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1: Georgian Med News. 2008 Apr;(157):39-44.

IgE-MEDIATED FOOD HYPERSENSITIVITY DISORDERS.

Gotua M, Lomidze N, Dolidze N, Gotua T.

Center of Allergy and Immunology.

Food allergy has become a serious health concern especially in developed
countries in the past two decades. In general population approximately 4-6% of
children and 1-3% of adults experience food allergy. The article reviews IgE -
mediated food hypersensitivity disorders. Epidemiology, Mechanism, Clinical
manifestations, Genetically modified crops (GMOs), Diagnosis, Prevention and
Treatment of IgE-mediated food allergies are discussed. The investigations show
that over 90% of IgE-mediated food allergies in childhood are caused by: cow's
milk, hen's egg, soy, peanuts, tree nuts, wheat, fish and shellfish. Also the
causes of food allergy are food additives, genetically modified crops. Risk
factors for food-dependant exercise-induced anaphylaxis include asthma and
previous allergic reactions to the causative food. Food allergy is one of the
most common causes of systematic anaphylaxis and anaphylactoid reactions, with an
annual incidence of four cases per million populations and estimated 500 deaths
annually. In addition to gastrointestinal symptoms, individuals may experience
urticaria, angioedema, atopic dermatitis, oral syndrome, asthma, rhinitis,
conjunctivitis, hypotension, shock and cardiac arrhythmias, caused by the massive
release of mediators from mast cells and basophiles. Diagnosis of food allergy is
based on history, detailed dietary analysis, skin testing, measuring specific IgE
in blood serum and challenge tests. Treatment and prevention includes: avoidance
diet, application of autoinjectable epinephrine, H1 and H2 antihistamines,
corticosteroids, antileukotriens, prostaglandin synthetase inhibitors, cromolyn
sodium, etc.

PMID: 18487689 [PubMed - in process]

2: Appl Environ Microbiol. 2008 May 16. [Epub ahead of print]

Effectiveness of Bt chickpeas and the entomopathogenic fungus Metarhizium
anisopliae to control Helicoverpa armigera (Lepidoptera: Noctuidae).

Lawo NC, Mahon RJ, Milner RJ, Sarmah BK, Higgins TJ, Romeis J.

Agroscope Reckenholz-Tö¤nikon Research Station ART, Reckenholzstr. 191, 8046
Zurich, Switzerland; CSIRO Entomology, GPO BOX 1700, Canberra, ACT 2601,
Australia; Department of Agricultural Biotechnology, Assam Agricultural
University, Jorhat 785 013, India; CSIRO Plant Industry, GPO Box 1600, Canberra
ACT 2601, Australia.

The use of genetically modified crops expressing lepidopteran-specific Cry
proteins derived from the soil bacterium Bacillus thuringiensis (Bt) is an
effective method to control the polyphagous pest Helicoverpa armigera. As H.
armigera potentially develops resistance to Cry proteins, Bt crops should be
regarded as one tool in integrated pest management. Therefore, they should be
compatible with biological control. Bioassays were conducted to understand the
interactions between a Cry2Aa-expressing chickpea line, either a susceptible or
Cry2A-resistant H. armigera strain, and the entomopathogenic fungus Metarhizium
anisopliae. In a first concentration-response assay, Cry2A-resistant larvae were
more tolerant to M. anisopliae than susceptible larvae, while in a second
bioassay the fungus caused similar mortalities to the two strains fed control
chickpea leaves. Thus, resistance to Cry2A did not cause any fitness costs that
would become visible in an increased susceptibility to the fungus. On Bt chickpea
leaves, susceptible H. armigera larvae were more sensitive to M. anisopliae than
on control leaves. It appeared that sublethal damage induced by the Bt-toxin
enhanced the effectiveness of M. anisopliae. For Cry2A-resistant larvae the
mortality caused by the fungus was similar when fed either food source. To
examine which strain would be more likely to be exposed to the fungus, their
movement on control and Bt chickpea plants was compared. Movement did not appear
to differ among larvae on Bt or conventional chickpea as indicated by the number
of leaflets damaged per leaf. The findings suggest that Bt chickpeas and M.
anisopliae are compatible to control H. armigera.

PMID: 18487396 [PubMed - as supplied by publisher]

3: Science. 2008 Apr 25;320(5875):473-5.

Is the drought over for pharming?

Kaiser J.

Publication Types:
News

PMID: 18436771 [PubMed - indexed for MEDLINE]

4: Science. 2008 Apr 25;320(5875):472.

Papaya takes on ringspot virus and wins.

Stokstad E.

Publication Types:
News

PMID: 18436770 [PubMed - indexed for MEDLINE]

5: Science. 2008 Apr 25;320(5875):468-71.

Tough lessons from golden rice.

Enserink M.

Publication Types:
News

PMID: 18436769 [PubMed - indexed for MEDLINE]

6: Science. 2008 Apr 25;320(5875):466-7.

GM crops: a world view.

[No authors listed]

Publication Types:
News

PMID: 18436768 [PubMed - indexed for MEDLINE]

7: Science. 2008 Apr 25;320(5875):465.

Multimedia feature: plant genomes.

[No authors listed]

Publication Types:
Interactive Tutorial

PMID: 18436767 [PubMed - indexed for MEDLINE]

8: Science. 2008 Apr 25;320(5875):465.

Green genes. Plant genomes. Introduction.

Zahn LM, Hines PJ, Pennisi E, Travis J.

Publication Types:
Introductory Journal Article

PMID: 18436766 [PubMed - indexed for MEDLINE]

9: Science. 2008 Apr 25;320(5875):452-3.

Ecology. Harvesting data from genetically engineered crops.

Marvier M, Carriö¨re Y, Ellstrand N, Gepts P, Kareiva P, Rosi-Marshall E,
Tabashnik BE, Wolfenbarger LL.

Environmental Studies Institute, Santa Clara University, Santa Clara, CA 95053,
USA. mmarvier@scu.edu

PMID: 18436759 [PubMed - indexed for MEDLINE]

10: Science. 2008 Apr 25;320(5875):425.

Seeds of a perfect storm.

Fedoroff N.

Publication Types:
Editorial

PMID: 18436745 [PubMed - indexed for MEDLINE]

11: J Nutr. 2008 May;138(5):921-6.

Lysozyme transgenic goats' milk influences gastrointestinal morphology in young
pigs.

Brundige DR, Maga EA, Klasing KC, Murray JD.

Department of Animal Science, University of California, Davis, CA 95616, USA.

Transgenesis provides a method of expressing novel proteins in milk to increase
the functional benefits of milk consumption. Transgenic goats expressing human
lysozyme (hLZ) at 67% of the concentration in human breast milk were produced,
thereby enhancing the antimicrobial properties of goats' milk. The objective of
this study was to investigate the impact of pasteurized milk containing hLZ on
growth, the intestinal epithelium, and an enteropathogenic Escherichia coli
(EPEC) infection in young weaned pigs. Pigs were placed into 4 groups and fed a
diet of solid food and either control (nontransgenic) goats' milk or milk from
hLZ-transgenic goats. Growth was assessed by weight gain. Nonchallenged pigs were
necropsied after 6 wk, whereas the remaining pigs were necropsied at 7 wk
following bacterial challenge. We determined the numbers of total coliforms and
E. coli and examined small intestinal histology for all pigs. Complete blood
counts were also determined pre- and postchallenge. Challenged pigs receiving hLZ
milk had fewer total coliforms (P = 0.029) and E. coli (P = 0.030) in the ileum
than controls. hLZ-fed pigs also had a greater duodenal villi width (P = 0.029)
than controls. Additionally, nonchallenged hLZ-fed pigs had fewer intraepithelial
lymphocytes per micron of villi height (P = 0.020) than nonchallenged controls.
These results indicate that the consumption of pasteurized hLZ goats' milk has
the potential to improve gastrointestinal health and is protective against an
EPEC in young weaned pigs. These same benefits may occur in young children if
they were to consume milk from hLZ-transgenic goats.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18424602 [PubMed - indexed for MEDLINE]

12: FEMS Microbiol Lett. 2008 Jun;283(1):62-8. Epub 2008 Apr 16.

Intragastric administration with recombinant Lactococcus lactis producing heme
oxygenase-1 prevents lipopolysaccharide-induced endotoxemia in rats.

Pang Q, Ji Y, Li Y, Bermöºdez-Humarö¡n LG, Hu G, Zeng Y.

Department of Pharmacology, Nanjing Medical University, Nanjing, Jiangsu, China.

Gut injury is a pivotal initiating event in the dysfunctional inflammatory
response that causes postinjury multiple organ failure. Heme oxygenase-1 (HO-1)
is an important enzyme that provides cellular protection against oxidative stress
in different in vitro and in vivo systems. In this study, we evaluated the
protective effects of intragastrically administered live Lactococcus lactis
secreting bioactive HO-1 to treat intestinal mucosal injury induced by
lipopolysaccharide in rats. Intragastric administration with this recombinant L.
lactis strain led to active delivery of HO-1 at the mucosa and significantly
decreased morbidity and mortality of lipopolysaccharide -induced endotoxemia as
confirmed by blinded macroscopic and microscopic inflammatory scores (Chiu's
grade), myeloperoxidase activity, mortality, and tumor necrosis factor-alpha and
IL-10 cytokine stimulation. This protective effect could be abolished by an HO-1
inhibitor, the zinc protoporphyrin-IX. Our results suggest that a food-grade
bacterium genetically modified to deliver bioactive HO-1 in situ exerts a
protective effect against intestinal mucosal injury in rats with endotoxemia via
modulation of the immune system. This novel approach may be beneficial for the
maintenance of the intestinal barrier and anti-inflammatory response of the lower
intestine.

PMID: 18422629 [PubMed - in process]

13: Science. 2008 Apr 18;320(5874):320-1.

Ecology. Agriculture at a crossroads.

Kiers ET, Leakey RR, Izac AM, Heinemann JA, Rosenthal E, Nathan D, Jiggins J.

Institute of Ecological Science, Faculty of Earth and Life Sciences, Vrije
Universiteit, De Boelelaan 1085, NL-1081 HV Amsterdam, Netherlands.
ekiers@falw.vu.nl

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18420917 [PubMed - indexed for MEDLINE]

14: Transgenic Res. 2008 Apr 11. [Epub ahead of print]

Environmental impact of herbicide regimes used with genetically modified
herbicide-resistant maize.

Devos Y, Cougnon M, Vergucht S, Bulcke R, Haesaert G, Steurbaut W, Reheul D.

Department of Plant Production, Faculty of Bioscience Engineering, Ghent
University, Coupure Links 653, 9000, Ghent, Belgium, Yann.Devos@UGent.be.

With the potential advent of genetically modified herbicide-resistant (GMHR)
crops in the European Union, changes in patterns of herbicide use are predicted.
Broad-spectrum, non-selective herbicides used with GMHR crops are expected to
substitute for a set of currently used herbicides, which might alter the
agro-environmental footprint from crop production. To test this hypothesis, the
environmental impact of various herbicide regimes currently used with non-GMHR
maize in Belgium was calculated and compared with that of possible herbicide
regimes applied in GMHR maize. Impacts on human health and the environment were
calculated through the pesticide occupational and environmental risk (POCER)
indicator. Results showed that the environmental impact of herbicide regimes
solely relying on the active ingredients glyphosate (GLY) or glufosinate-ammonium
(GLU) is lower than that of herbicide regimes applied in non-GMHR maize. Due to
the lower potential of GLY and GLU to contaminate ground water and their lower
acute toxicity to aquatic organisms, the POCER exceedence factor values for the
environment were reduced approximately by a sixth when GLY or GLU is used alone.
However, the environmental impact of novel herbicide regimes tested may be
underestimated due to the assumption that active ingredients used with GMHR maize
would be used alone. Data retrieved from literature suggest that weed control
efficacy is increased and resistance development delayed when GLY or GLU is used
together with other herbicides in the GMHR system. Due to the partial instead of
complete replacement of currently used herbicide regimes, the beneficial
environmental impact of novel herbicide regimes might sometimes be reduced or
counterbalanced. Despite the high weed control efficacy provided by the
biotechnology-based weed management strategy, neither indirect harmful effects on
farmland biodiversity through losses in food resources and shelter, nor shifts in
weed communities have been demonstrated in GMHR maize yet. However, with the
increasing adoption rate of GMHR maize and their associated novel herbicide
regimes, this situation is expected to change in the short-term.

PMID: 18404410 [PubMed - as supplied by publisher]

15: Science. 2008 Apr 11;320(5873):171-3.

Plant genetics. The blue revolution, drop by drop, gene by gene.

Pennisi E.

Publication Types:
News

PMID: 18403686 [PubMed - indexed for MEDLINE]

16: Food Chem Toxicol. 2008 Jun;46(6):2201-13. Epub 2008 Feb 29.

Subchronic feeding study of herbicide-tolerant soybean DP-356ö˜43-5 in
Sprague-Dawley rats.

Appenzeller LM, Munley SM, Hoban D, Sykes GP, Malley LA, Delaney B.

Pioneer Hi-Bred International, Inc., Johnston, IA, USA.

Optimumtrade markGATtrade mark(1) soybean is a genetically modified (GM) soybean
containing event DP-356ö˜43-5 (356043) that was produced by integration of the
coding sequences of the GAT4601 and GM-HRA proteins. In planta expression of
these proteins confers tolerance to glyphosate and sulfonylurea/imidazolinone
herbicides, respectively. This paper reports the results from a subchronic rat
feeding study conducted with 356043 soybeans. Dehulled/defatted toasted meal and
toasted ground hulls were prepared from soybeans from untreated plants (356043),
herbicide-treated plants (356043+Gly/SU), non-transgenic isoline control (091),
and three commercial non-transgenic reference varieties (93B86, 93B15, and
93M40). Individual diets conforming to standard certified rodent chow formulation
(Purina Rodent LabDiet((R)) 5002) were prepared with 20% meal (w/w) and 1.5%
hulls (w/w). Diets were fed to young adult Sprague-Dawley rats (12/sex/group) for
at least 93 days. Compared with rats fed the isoline control or conventional
reference diets, no biologically-relevant, adverse effects were observed in rats
fed diets containing 356043 or 356043+Gly/SU soybean with respect to body
weight/gain, food consumption/efficiency, clinical signs, mortality,
ophthalmology, neurobehavioral assessments (sensory response, grip strength,
motor activity), clinical pathology (hematology, coagulation, serum chemistry,
urinalysis), organ weights, and gross and microscopic pathology. The results from
this study indicate that 356043 soybeans are as safe and nutritious as
conventional non-GM soybeans.

PMID: 18403083 [PubMed - in process]

17: Nature. 2008 Mar 6;452(7183):122-4.

Almost in bloom.

Marris E.

PMID: 18396501 [PubMed - indexed for MEDLINE]

18: Nat Biotechnol. 2008 Apr;26(4):384-6.

Comment on:
Nat Biotechnol. 2008 Mar;26(3):260.

Bt corn in Spain--the performance of the EU's first GM crop.

Gö¨mez-Barbero M, Berbel J, Rodrö­guez-Cerezo E.

Publication Types:
Comment
Letter
Research Support, Non-U.S. Gov't

PMID: 18392015 [PubMed - indexed for MEDLINE]

19: Nat Biotechnol. 2008 Apr;26(4):379; discussion 379-80.

Comment on:
Nat Biotechnol. 2007 Dec;25(12):1330.

An inconvenient version of events.

Monastra G.

Publication Types:
Comment
Letter

PMID: 18392011 [PubMed - indexed for MEDLINE]

20: Nat Biotechnol. 2008 Apr;26(4):365.

Tear-free onions.

Aldridge S.

Publication Types:
News

PMID: 18392004 [PubMed - indexed for MEDLINE]

21: Br J Nutr. 2008 Feb;99 Suppl 1:S22-5.

Influence of parental attitudes in the development of children eating behaviour.

Scaglioni S, Salvioni M, Galimberti C.

Pediatric Clinic S. Paolo Hospital University of Milan, Milan, Italy.
silviascaglioni@unimi.it

The present paper is a review of available data on effects of parental feeding
attitudes and styles on child nutritional behaviour. Food preferences develop
from genetically determined predispositions to like sweet and salty flavours and
to dislike bitter and sour tastes. There is evidence for existence of some
innate, automatic mechanism that regulate appetite. However, from birth genetic
predispositions are modified by experience. There are mechanisms of taste
development: mere exposure, medicine effect, flavour learning, flavour nutrient
learning. Parents play a pivotal role in the development of their child's food
preferences and energy intake, with research indicating that certain child
feeding practices, such as exerting excessive control over what and how much
children eat, may contribute to childhood overweight. Mothers are of particular
interest on children's eating behaviour, as they have been shown to spend
significantly more time than fathers in direct interactions with their children
across several familial situations.A recent paper describes two primary aspects
of control: restriction, which involves restricting children's access to junk
foods and restricting the total amount of food, and pressure, which involves
pressuring children to eat healthy foods (usually fruits and vegetables) and
pressuring to eat more in general.The results showed significant correlations
between parent and child for reported nutritional behaviour like food intake,
eating motivations, and body dis- and satisfaction. Parents create environments
for children that may foster the development of healthy eating behaviours and
weight, or that may promote overweight and aspects of disordered eating. In
conclusion positive parental role model may be a better method for improving a
child's diet than attempts at dietary control.

PMID: 18257948 [PubMed - in process]

22: BMC Biotechnol. 2008 Apr 3;8:36.

Development of patatin knockdown potato tubers using RNA interference (RNAi)
technology, for the production of human-therapeutic glycoproteins.

Kim YS, Lee YH, Kim HS, Kim MS, Hahn KW, Ko JH, Joung H, Jeon JH.

Plant Genome Research Center, KRIBB, Daejeon 305-806, Korea. yoon1920@kribb.re.kr

BACKGROUND: Patatins encoded by a multi-gene family are one of the major storage
glycoproteins in potato tubers. Potato tubers have recently emerged as
bioreactors for the production of human therapeutic glycoproteins (vaccines).
Increasing the yield of recombinant proteins, targeting the produced proteins to
specific cellular compartments, and diminishing expensive protein purification
steps are important research goals in plant biotechnology. In the present study,
potato patatins were eliminated almost completely via RNA interference (RNAi)
technology to develop potato tubers as a more efficient protein expression
system. The gene silencing effect of patatins in the transgenic potato plants was
examined at individual isoform levels. RESULTS: Based upon the sequence
similarity within the multi-gene family of patatins, a highly conserved target
sequence (635 nts) of patatin gene pat3-k1 [GenBank accession no. DQ114421] in
potato plants (Solanum tuberosum L.) was amplified for the construction of a
patatin-specific hairpin RNAi (hpRNAi) vector. The CaMV 35S promoter-driven
patatin hpRNAi vector was transformed into the potato cultivar Desiree by
Agrobacterium-mediated transformation. Ten transgenic potato lines bearing
patatin hpRNA were generated. The effects of RNA interference were characterized
at both the protein and mRNA levels using 1D and 2D SDS/PAGE and quantitative
real-time RT-PCR analysis. Dependent upon the patatin hpRNAi line, patatins
decreased by approximately 99% at both the protein and mRNA levels. However, the
phenotype (e.g. the number and size of potato tuber, average tuber weight, growth
pattern, etc.) of hpRNAi lines was not distinguishable from wild-type potato
plants under both in vitro and ex vitro growth conditions. During glycoprotein
purification, patatin-knockdown potato tubers allowed rapid purification of other
potato glycoproteins with less contamination of patatins. CONCLUSION:
Patatin-specific hpRNAi effectively suppressed the expression of a majority of
patatin variants in potato tubers via the specific degradation of individual
mRNAs of the patatin multi-gene family. More importantly, patatin-knockdown
potato tubers appear to be an ideal host for the production of human therapeutic
glycoproteins, because they eventually allow fast, easy purification of
recombinant proteins, with less contamination from potato glycoprotein patatins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18384693 [PubMed - indexed for MEDLINE]

23: Rev Med Suisse. 2008 Jan 30;4(142):314.

[Food and milk clones declared]

[Article in French]

Nau JY.

PMID: 18383942 [PubMed - indexed for MEDLINE]

24: J Am Vet Med Assoc. 2008 Mar 1;232(5):667.

FDA reaches final conclusion that food from clones is safe.

[No authors listed]

Publication Types:
News

PMID: 18380061 [PubMed - indexed for MEDLINE]

25: Proc Natl Acad Sci U S A. 2008 Apr 1;105(13):5129-33. Epub 2008 Mar 28.

Anthropogenic increase in carbon dioxide compromises plant defense against
invasive insects.

Zavala JA, Casteel CL, Delucia EH, Berenbaum MR.

Institute for Genomic Biology and Departments of Plant Biology and Entomology,
University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

Elevated levels of atmospheric carbon dioxide (CO2), a consequence of
anthropogenic global change, can profoundly affect the interactions between crop
plants and insect pests and may promote yet another form of global change: the
rapid establishment of invasive species. Elevated CO2 increased the
susceptibility of soybean plants grown under field conditions to the invasive
Japanese beetle (Popillia japonica) and to a variant of western corn rootworm
(Diabrotica virgifera virgifera) resistant to crop rotation by down-regulating
gene expression related to defense signaling [lipoxygenase 7 (lox7), lipoxygenase
8 (lox8), and 1-aminocyclopropane-1-carboxylate synthase (acc-s)]. The
down-regulation of these genes, in turn, reduced the production of cysteine
proteinase inhibitors (CystPIs), which are specific deterrents to coleopteran
herbivores. Beetle herbivory increased CystPI activity to a greater degree in
plants grown under ambient than under elevated CO2. Gut cysteine proteinase
activity was higher in beetles consuming foliage of soybeans grown under elevated
CO2 than in beetles consuming soybeans grown in ambient CO2, consistent with
enhanced growth and development of these beetles on plants grown in elevated CO2.
These findings suggest that predicted increases in soybean productivity under
projected elevated CO2 levels may be reduced by increased susceptibility to
invasive crop pests.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18375762 [PubMed - indexed for MEDLINE]

26: Biochem Biophys Res Commun. 2008 May 30;370(2):344-7. Epub 2008 Mar 26.

Enhancement of disease resistance to Magnaporthe grisea in rice by accumulation
of hydroxy linoleic acid.

Yara A, Yaeno T, Montillet JL, Hasegawa M, Seo S, Kusumi K, Iba K.

Department of Biology, Faculty of Sciences, Kyushu University, 6-10-1 Hakozaki,
Higashi-ku, Fukuoka 812-8581, Japan.

Linoleic acid (18:2) and linolenic acid (18:3) are sources for various oxidized
metabolites called oxylipins, some of which inhibit growth of fungal pathogens.
In a previous study, we found disease resistance to rice blast fungus Magnaporthe
grisea enhanced in 18:2-accumulating transgenic rice (F78Ri) in which the
conversion from 18:2 to 18:3 was suppressed. Here, we demonstrate that
18:2-derived hydroperoxides and hydroxides (HPODEs and HODEs, respectively)
inhibit growth of M. grisea more strongly than their 18:3-derived counterparts.
Furthermore, in F78Ri plants, the endogenous levels of HPODEs and HODEs increased
significantly, compared with wild-type plants. These results suggest that the
increased accumulation of antifungal oxylipins, such as HPODEs and HODEs, causes
the enhancement of disease resistance against M. grisea.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18373976 [PubMed - indexed for MEDLINE]

27: RNA. 2008 May;14(5):903-13. Epub 2008 Mar 26.

Hairpin RNAs derived from RNA polymerase II and polymerase III promoter-directed
transgenes are processed differently in plants.

Wang MB, Helliwell CA, Wu LM, Waterhouse PM, Peacock WJ, Dennis ES.

Commonwealth Scientific and Industrial Research Organisation Plant Industry,
Canberra, Australian Capital Territory 2601, Australia. ming-bo.wang@csiro.au

RNA polymerase III (Pol III) as well as Pol II (35S) promoters are able to drive
hairpin RNA (hpRNA) expression and induce target gene silencing in plants. siRNAs
of 21 nt are the predominant species in a 35S Pol II line, whereas 24- and/or
22-nucleotide (nt) siRNAs are produced by a Pol III line. The 35S line
accumulated the loop of the hpRNA, in contrast to full-length hpRNA in the Pol
III line. These suggest that Pol II and Pol III-transcribed hpRNAs are processed
by different pathways. One Pol III transgene produced only 24-nt siRNAs but
silenced the target gene efficiently, indicating that the 24-nt siRNAs can direct
mRNA degradation; specific cleavage was confirmed by 5' rapid amplification of
cDNA ends (RACE). Both Pol II- and Pol III-directed hpRNA transgenes induced
cytosine methylation in the target DNA. The extent of methylation is not
correlated with the level of 21-nt siRNAs, suggesting that they are not effective
inducers of DNA methylation. The promoter of a U6 transgene was significantly
methylated, whereas the promoter of the endogenous U6 gene was almost free of
cytosine methylation, suggesting that endogenous sequences are more resistant to
de novo DNA methylation than are transgene constructs.

PMID: 18367720 [PubMed - indexed for MEDLINE]

28: J Infect Dis. 2008 Feb 15;197 Suppl 1:S25-8.

Genetic strategy to prevent influenza virus infections in animals.

Chen J, Chen SC, Stern P, Scott BB, Lois C.

Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, MA
02139, USA. jchen@mit.edu

The natural reservoirs of influenza viruses are aquatic birds. After adaptation,
avian viruses can acquire the ability to infect humans and cause severe disease.
Because domestic poultry serves as a key link between the natural reservoir of
influenza viruses and epidemics and pandemics in human populations, an effective
measure to control influenza would be to eliminate or reduce influenza virus
infection in domestic poultry. The development and distribution of
influenza-resistant poultry represents a proactive strategy for controlling the
origin of influenza epidemics and pandemics in both poultry and human
populations. Recent developments in RNA interference and transgenesis in birds
should facilitate the development of influenza-resistant poultry.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 18269324 [PubMed - indexed for MEDLINE]

29: Genome. 2008 Jan;51(1):41-9.

An efficient field screening procedure for identifying transposants for
constructing an Ac/Ds-based insertional-mutant library of rice.

Luan WJ, He CK, Hu GC, Dey M, Fu YP, Si HM, Zhu L, Liu WZ, Duan F, Zhang H, Liu
WY, Zhuo RY, Garg A, Wu R, Sun ZX.

China National Rice Research Institute, State Key Laboratory of Rice Biology,
Hangzhou, Zhejiang, People's Republic of China.

An efficient system was developed, and several variables tested, for generating a
large-scale insertional-mutagenesis population of rice. The most important
feature in this improved Ac/Ds tagging system is that one can conveniently carry
out large-scale screening in the field and select transposants at the seedling
stage. Rice was transformed with a plasmid that includes a Basta-resistance gene
(bar). After the Ds element is excised during transposition, bar becomes adjacent
to the ubiquitin promoter, and the rice plant becomes resistant to the herbicide
Basta. In principle, one can plant up to one million plants in the field and
select those plants that survive after spraying with Basta. To test the utility
of this system, 4 Ds starter lines were crossed with 14 different Ac plants, and
many transposants were successfully identified after planting 134,285 F2 plants
in the field. Over 2,800 of these transposants were randomly chosen for PCR
analysis, and the results fully confirmed the reliability of the field screening
procedure.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18356938 [PubMed - indexed for MEDLINE]

30: EMBO J. 2008 Apr 23;27(8):1183-96. Epub 2008 Mar 20.

Regulation of endocytic recycling by C. elegans Rab35 and its regulator RME-4, a
coated-pit protein.

Sato M, Sato K, Liou W, Pant S, Harada A, Grant BD.

Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway,
NJ, USA. sato-ken@showa.gunma-u.ac.jp

Using Caenorhabditis elegans genetic screens, we identified receptor-mediated
endocytosis (RME)-4 and RME-5/RAB-35 as important regulators of yolk endocytosis
in vivo. In rme-4 and rab-35 mutants, yolk receptors do not accumulate on the
plasma membrane as would be expected in an internalization mutant, rather the
receptors are lost from cortical endosomes and accumulate in dispersed small
vesicles, suggesting a defect in receptor recycling. Consistent with this,
genetic tests indicate the RME-4 and RAB-35 function downstream of clathrin,
upstream of RAB-7, and act synergistically with recycling regulators RAB-11 and
RME-1. We find that RME-4 is a conserved DENN domain protein that binds to RAB-35
in its GDP-loaded conformation. GFP-RME-4 also physically interacts with AP-2, is
enriched on clathrin-coated pits, and requires clathrin but not RAB-5 for
cortical association. GFP-RAB-35 localizes to the plasma membrane and early
endocytic compartments but is lost from endosomes in rme-4 mutants. We propose
that RME-4 functions on coated pits and/or vesicles to recruit RAB-35, which in
turn functions in the endosome to promote receptor recycling.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 18354496 [PubMed - indexed for MEDLINE]

31: Nature. 2008 Mar 20;452(7185):273-7.

Water: more crop per drop.

Marris E.

Publication Types:
News

PMID: 18354452 [PubMed - indexed for MEDLINE]

32: J Neurosci. 2008 Mar 19;28(12):3103-13.

Rapid consolidation to a radish and protein synthesis-dependent long-term memory
after single-session appetitive olfactory conditioning in Drosophila.

Krashes MJ, Waddell S.

Department of Neurobiology, University of Massachusetts Medical School,
Worcester, Massachusetts 01605, USA.

In Drosophila, formation of aversive olfactory long-term memory (LTM) requires
multiple training sessions pairing odor and electric shock punishment with rest
intervals. In contrast, here we show that a single 2 min training session pairing
odor with a more ethologically relevant sugar reinforcement forms long-term
appetitive memory that lasts for days. Appetitive LTM has some mechanistic
similarity to aversive LTM in that it can be disrupted by cycloheximide, the
dCreb2-b transcriptional repressor, and the crammer and tequila LTM-specific
mutations. However, appetitive LTM is completely disrupted by the radish mutation
that apparently represents a distinct mechanistic phase of consolidated aversive
memory. Furthermore, appetitive LTM requires activity in the dorsal paired medial
neuron and mushroom body alpha'beta' neuron circuit during the first hour after
training and mushroom body alphabeta neuron output during retrieval, suggesting
that appetitive middle-term memory and LTM are mechanistically linked. Last,
experiments feeding and/or starving flies after training reveals a critical
motivational drive that enables appetitive LTM retrieval.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 18354013 [PubMed - indexed for MEDLINE]

33: Environ Entomol. 2008 Feb;37(1):263-70.

Tri-trophic interactions between Bt cotton, the herbivore Aphis gossypii Glover
(Homoptera: Aphididae), and the predator Chrysopa pallens (Rambur) (Neuroptera:
Chrysopidae).

Guo JY, Wan FH, Dong L, Lö¶vei GL, Han ZJ.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of
Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094,
China. guojy@cjac.org.cn

Tri-trophic impacts of transgenic Bacillus thuringiensis (Bt) cotton GK12 and
NuCOTN 99B were studied using a predator, the great lacewing Chrysopa pallens
(Rambur), and its prey, the cotton aphid Aphis gossypii Glover, in laboratory
feeding experiments. The parental nontransgenic cotton cultivar of GK12 was used
as control. The predator was fed with uniform (aphids from a single cultivar) or
mixed prey (aphids from the three cotton cultivars provided on alternate days).
Mortality and development of the immature stages, pupal body mass, adult sex
ratio, fecundity, and egg viability of C. pallens were measured. When fed
GK12-originated aphid prey, pupal body mass of C. pallens was significantly
higher than that of the control, more females emerged, and these females laid
significantly more eggs. Other parameters were not impacted. Females emerging
from larvae maintained on NuCOTN 99B-originated prey laid fewer eggs than those
maintained on GK12. Other measurements did not differ significantly between the
two Bt cotton cultivars. Compared with the control, mixed feeding significantly
prolonged pupal development time and increased pupal body mass and percentage of
females but did not affect other parameters. These results indicate that C.
pallens is sensitive to aphid prey from different cotton cultivars. Transgenic Bt
cotton GK12-originated aphid prey has no adverse impact on survival, development,
and fecundity of C. pallens. Between the two Bt cotton cultivars, NuCOTN
99B-originated aphid prey provided to C. pallens in the larval stage may lower
female fecundity. Mixed feeding of C. pallens with the two Bt cotton-originated
prey and non-Bt prey may have some adverse impacts on pupal development.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 18348819 [PubMed - indexed for MEDLINE]

34: Environ Entomol. 2008 Feb;37(1):255-62.

Cry1F protein not detected in soil after three years of transgenic Bt corn (1507
corn) use.

Shan G, Embrey SK, Herman RA, McCormick R.

Dow AgroSciences, 9330 Zionsville Rd., Indianapolis, IN 46268, USA. gshan@dow.com

To evaluate the potential of Bacillus thuringiensis (Bt) Cry1F protein
accumulation in soil, transgenic corn containing event DAS-01507-1 encoding the
cry1F gene was grown in three field sites for 3 consecutive yr, and the corn
plants were incorporated into the soil through postseason tillage or no tillage
each year. Soil samples were collected from these fields, and the level of Cry1F
protein in these samples was determined using an enzyme-linked immunosorbent
assay (ELISA) with a synthetic invertebrate gut fluid as an extraction buffer.
The ELISA was validated in soil matrices over the concentration range of 18-180
ng/g dry weight, with a limit of detection of 4.5 ng/g dry weight. The assay was
shown to have good accuracy and precision. No detectable Cry1F protein was found
in any of the soil samples collected from the Cry1F corn fields. Soil also was
bioassayed, and no biological activity was observed against Heliothis virescens
neonates. These results indicate that the level of Cry1F protein accumulated in
soil after 3-yr continuous planting of transgenic Cry1F corn is negligible.

PMID: 18348818 [PubMed - indexed for MEDLINE]

35: Environ Entomol. 2008 Feb;37(1):1-10.

Selection of nontarget arthropod taxa for field research on transgenic
insecticidal crops: using empirical data and statistical power.

Prasifka JR, Hellmich RL, Dively GP, Higgins LS, Dixon PM, Duan JJ.

USDA-ARS, Corn Insects and Crop Genetics Research Unit, Genetics Laboratory c/o
Insectary, Iowa State University, Ames, IA 50011, USA. prasifka@iastate.edu

One of the possible adverse effects of transgenic insecticidal crops is the
unintended decline in the abundance of nontarget arthropods. Field trials
designed to evaluate potential nontarget effects can be more complex than
expected because decisions to conduct field trials and the selection of taxa to
include are not always guided by the results of laboratory tests. Also, recent
studies emphasize the potential for indirect effects (adverse impacts to
nontarget arthropods without feeding directly on plant tissues), which are
difficult to predict because of interactions among nontarget arthropods, target
pests, and transgenic crops. As a consequence, field studies may attempt to
monitor expansive lists of arthropod taxa, making the design of such broad
studies more difficult and reducing the likelihood of detecting any negative
effects that might be present. To improve the taxonomic focus and statistical
rigor of future studies, existing field data and corresponding power analysis may
provide useful guidance. Analysis of control data from several nontarget field
trials using repeated-measures designs suggests that while detection of small
effects may require considerable increases in replication, there are taxa from
different ecological roles that are sampled effectively using standard methods.
The use of statistical power to guide selection of taxa for nontarget trials
reflects scientists' inability to predict the complex interactions among
arthropod taxa, particularly when laboratory trials fail to provide guidance on
which groups are more likely to be affected. However, scientists still may
exercise judgment, including taxa that are not included in or supported by power
analyses.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18348790 [PubMed - indexed for MEDLINE]

36: Arch Environ Contam Toxicol. 2008 Mar 18. [Epub ahead of print]

Reduced Fitness of Daphnia magna Fed a Bt-Transgenic Maize Variety.

Bö¸hn T, Primicerio R, Hessen DO, Traavik T.

Genö¸kÁ€”Centre for Biosafety, The Science Park, P.O. Box 6418, Tromso, 9294,
Norway, thomas@genok.org.

Genetically modified (GM) maize expressing the Bt-toxin Cry1Ab (Bt-maize) was
tested for effects on survival, growth, and reproduction of the water flea
Daphnia magna, a crustacean arthropod commonly used as a model organism in
ecotoxicological studies. In three repeated experiments, D. magna were fed 100%
ground maize in suspension, using either GM or isogenic unmodified (UM) maize. D.
magna fed GM-maize showed a significantly reduced fitness performance: The
mortality was higher, a lower proportion of females reached sexual maturation,
and the overall egg production was lower compared to D. magna fed UM isogenic
maize. We conclude that the tested variety of Bt-maize and its UM counterpart do
not have the same quality as food sources for this widely used model organism.
The combination of a reduced fitness performance combined with earlier onset of
reproduction of D. magna fed Bt-maize indicates a toxic effect rather than a
lower nutritional value of the GM-maize.

PMID: 18347840 [PubMed - as supplied by publisher]

37: Medicina (Kaunas). 2008;44(2):87-99.

Genetically modified organisms: do the benefits outweigh the risks?

Hug K.

K. Hug, Department of Medical Ethics, Lund University, BMC C 13, 221 84 Lund,
Sweden. Kristina.Hug@med.lu.se

The objective of this literature review is to analyze the implications of using
genetically modified organisms (GMOs) as well as international and European
position regarding such organisms. METHOD: Review of international and European
legal requirements and ethical guidelines and relevant publications, found and
accessed with the help of PubMed and Lund University Library databases. RESULTS:
The article discusses the main application areas of GMOs, the expansion of using
GMOs in the world as well as the advantages and disadvantages of the implications
of their usage. It further provides an overview of the suggested ways to tackle
or avoid the GMO-related risks. The international and European positions
regarding the application of GMOs are discussed and European Directives,
Regulations, and ethical guidelines are overviewed. The article further presents
the public attitudes towards GMOs in Europe as well as overviews surveys
conducted at the national level. CONCLUSION: Suggested steps to tackle the
challenge of developing and managing biotechnology for the benefit of public
health and the environment are presented.

Publication Types:
Comparative Study
Review

PMID: 18344661 [PubMed - indexed for MEDLINE]

38: Shokuhin Eiseigaku Zasshi. 2008 Feb;49(1):16-22.

Development of event-specific quantitation method for GA21 maize, which is a gm
event without CaMV35S promoter.

Oguchi T, Onishi M, Chikagawa Y, Minegishi Y, Kodama T, Akiyama H, Ohno Y, Futo
S, Hino A, Furui S, Kitta K.

National Agriculture and Food Research Organization, National Food Research
Institute, Ibaraki, Japan.

A real-time PCR detection method was developed for event-specific quantitation of
Roundup Ready maize, GA21. The developed PCR method was designed to amplify an
artificial junction site between the native maize genome DNA and the recombinant
DNA of GA21 maize, which provides only one target sequence per haploid of GA21
genome. Thus, the amplification efficiency of the event-specific target for GA21
became closely similar to the amplification of SSIIb, and the conversion factor
(Cf) for the quantitation method was similar to the theoretical value. The
developed method demonstrated better performance than the existing
construct-specific method that has been used as a Japanese official method. The
developed method can easily be combined with the real-time PCR targeting of the
CaMV35S promoter, and the multiplexed method should be an effective screening
method for GM maize.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18344654 [PubMed - indexed for MEDLINE]

39: Science. 2008 Mar 14;319(5869):1474-6.

Erratum in:
Science. 2008 Apr 4;320(5872):50. Science.2008 Apr 18;320(5874):316.

Agriculture. Dueling visions for a hungry world.

Stokstad E.

Publication Types:
News

PMID: 18339914 [PubMed - indexed for MEDLINE]

40: Appetite. 2008 Jul;51(1):129-36. Epub 2008 Feb 7.

Acceptance of genetically modified foods: The relation between technology and
evaluation.

Tenbö¼lt P, De Vries NK, van Breukelen G, Dreezens E, Martijn C.

Department of Health Education and Promotion, Universiteit Maastricht, The
Netherlands.

This study investigates why consumers accept different genetically modified food
products to different extents. The study shows that whether food products are
genetically modified or not and whether they are processed or not are the two
important features that affect the acceptance of food products and their
evaluation (in terms of perceived healthiness, naturalness, necessity and
tastiness). The extent to which these evaluation attributes and acceptance of a
product are affected by genetic modification or processing depends on whether the
product is negatively affected by the other technology: Any technological change
to a 'natural' product (when nonprocessed products are genetically modified or
when non-genetically modified products are processed) affect evaluation and
acceptance stronger than a change to an technologically adapted product (when
processed products are also genetically modified or vice versa). Furthermore,
evaluation attributes appear to mediate the effects of genetic modification and
processing on acceptance.

PMID: 18336952 [PubMed - in process]

41: Environ Toxicol Chem. 2008 Apr;27(4):793-8.

Aquatic fate and effects of Bacillus thuringiensis Cry3Bb1 protein: toward risk
assessment.

Prihoda KR, Coats JR.

Pesticide Toxicology Laboratory, Department of Entomology, Iowa State University,
Ames, Iowa 50011, USA.

Genetically engineered crops expressing Bacillus thuringiensis (Bt) insecticidal
crystalline (Cry) proteins became commercially available in the United States in
1996. In 2006, 19 million ha of Bt corn were planted worldwide, which represents
a 10 million ha increase in 10 years. The sustainability of Bt crops is
important, because their use has significantly reduced the amount of chemical
insecticides necessary to control agricultural pests. Despite the high adoption
rates of this novel insecticide, little is known about the aquatic fate of
transgenic Bt proteins and their nontarget effects on aquatic invertebrates,
although several potential routes exist for their transport to aquatic systems.
Methods were developed to investigate the aquatic fate of transgenic Bt proteins
and to determine their potential effects on nontarget aquatic invertebrates.
Laboratory microcosms containing pond water only or pond water and sediment were
used to examine the fate of the coleopteran-active Bt Cry3Bb1 protein in
decomposing transgenic corn event MON863 (hereafter referred to as MON863 corn)
leaf, stalk, and root. A half-life of less than 3 d was found for Bt Cry3Bb1 from
decomposing MON863 corn residue. No Bt Cry3Bb1 was measured in the pond water or
sediment extracts of microcosms containing MON863 corn. In an acute, static,
partial-renewal toxicity test, Bt Cry3Bb1 protein from MON863 root extracts was
fed to Chironomus dilutus larvae for 10 d. A significant decrease in C. dilutus
survival at nominal concentrations of 30 ng/ml was found; however, no effect on
growth among the surviving larvae was observed.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18333682 [PubMed - indexed for MEDLINE]

42: Proc Natl Acad Sci U S A. 2008 Mar 18;105(11):4495-500. Epub 2008 Mar 10.

Integration of light and abscisic acid signaling during seed germination and
early seedling development.

Chen H, Zhang J, Neff MM, Hong SW, Zhang H, Deng XW, Xiong L.

Donald Danforth Plant Science Center, St Louis, MO 63132, USA.

Seed germination is regulated by endogenous hormonal cues and external
environmental stimuli such as water, low temperature, and light. After
germination, the young seedling must rapidly establish its root system and the
photoautotrophic capability appropriate to its surrounding environment. Light and
the phytohormone abscisic acid (ABA) both regulate seed germination and seedling
development, although how light and ABA signals are integrated at the molecular
level is not understood. Here, we found that the previously described
light-signaling component HY5 also mediates ABA response in seed germination,
early seedling growth, and root development in Arabidopsis. HY5 binds to the
promoter of the transcription factor ABI5 gene with high affinity and is required
for the expression of ABI5 and ABI5-targeted late embryogenesis-abundant genes in
seeds. Chromatin immunoprecipitation also indicated that the binding of HY5 to
the ABI5 promoter is significantly enhanced by ABA. Overexpression of ABI5
restores ABA sensitivity in hy5 and results in enhanced light responses and
shorter hypocotyls in the wild type. Our studies identified an unexpected mode of
light and ABA signal integration that may help young seedlings better adapt to
environmental stresses.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18332440 [PubMed - indexed for MEDLINE]

43: J Econ Entomol. 2008 Feb;101(1):182-9.

Effect of transgenic Bacillus thuringiensis rice lines on mortality and feeding
behavior of rice stem borers (Lepidoptera: Crambidae).

Chen H, Zhang G, Zhang Q, Lin Y.

National Key Laboratory of Crop Genetic Improvement and National Centre of Plant
Gene Research, Huazhong Agricultural University, Wuhan, 430070, China.

Ten transgenic Bacillus thuringiensis Bt rice, Oryza sativa L., lines with
different Bt genes (two Cry1Ac lines, three Cry2A lines, and five Cry9C lines)
derived from the same variety Minghui 63 were evaluated in both the laboratory
and the field. Bioassays were conducted by using the first instars of two main
rice lepidopteran insect species: yellow stem borer, Scirpophaga incertulas
(Walker) and Asiatic rice borer, Chilo suppressalis (Walker). All transgenic
lines exhibited high toxicity to these two rice borers. Field evaluation results
also showed that all transgenic lines were highly insect resistant with both
natural infestation and manual infestation of the neonate larvae of S. incertulas
compared with the nontransformed Minghui63. Bt protein concentrations in leaves
of 10 transgenic rice lines were estimated by the sandwich enzyme-linked
immunosorbent assay. The cry9C gene had the highest expression level, next was
cry2A gene, and the cry1Ac gene expressed at the lowest level. The feeding
behavior of 7-d-old Asiatic rice borer to three classes of Bt transgenic rice
lines also was detected by using rice culm cuttings. The results showed that
7-d-old larvae of Asiatic rice borer have the capacity to distinguish Bt and
non-Bt culm cuttings and preferentially fed on non-Bt cuttings. When only Bt culm
cuttings with three classes of different Bt proteins (CrylAc, Cry2A, and Cry9C)
were fed, significant distribution difference of 7-d-old Asiatic rice borer in
culm cuttings of different Bt proteins also was found. In the current study, we
evaluate different Bt genes in the same rice variety in both the laboratory and
the field, and also tested feeding behavior of rice insect to these Bt rice.
These data are valuable for the further development of two-toxin Bt rice and
establishment of appropriate insect resistance management in the future.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18330134 [PubMed - indexed for MEDLINE]

44: J Econ Entomol. 2008 Feb;101(1):56-60.

Flight take-off performance of Colorado potato beetle in relation to potato
phenology.

Mbungu NT, Boiteau G.

Department of Natural Resource Sciences, Macdonald Campus of McGill University,
21,111 Lakeshore Blvd., Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada.

The flight take-off frequency of adult Colorado potato beetles, Leptinotarsa
decemlineata (Say), from potato plants, Solanum tuberosum L. 'Red Pontiac' at the
bloom stage of development was 2.2-2.5-fold that of Colorado potato beetle from
plants at the vegetative stage. Tests were conducted in a flight chamber over a
period of 3 h. Prefeeding Colorado potato beetles for 48 h on potato plants at
the bloom or at the vegetative stage before placing them into the flight chamber
resulted in the same significantly higher flight take-off frequency from potato
plants at the bloom stage than from plants at the vegetative stage. These results
demonstrate that the factor in potato plants in bloom that stimulates the flight
take-off of the Colorado potato beetle is independent of the feeding history of
the beetles and begins acting only when the beetles are in the presence of the
plant. According to these results, the dispersal of adult Colorado potato beetles
from potato fields in bloom to younger potato fields with plants at the
vegetative stage, previously reported in the literature, is at least partly
explained by the effect of plant phenology on the frequency of flight take-off.
Results confirm the value of planting potato fields of similar phenology over as
wide an area as possible to reduce Colorado potato beetle dispersal between
fields. Results also imply that staggering the planting dates of conventional
potato refuge areas near Colorado potato beetle transgenic or conventionally
resistant potato fields is a sound management practice, because it promotes the
movement of wild beetles over to the adjacent younger resistant crops.

PMID: 18330116 [PubMed - indexed for MEDLINE]

45: Food Chem Toxicol. 2008 Mar;46S1:S2-S70. Epub 2008 Feb 13.

Safety and nutritional assessment of GM plants and derived food and feed: The
role of animal feeding trials.

Report of the EFSA GMO Panel Working Group on Animal Feeding Trials.

Adopted by the Scientific Panel on Genetically Modified Organisms(2) on 12
September 2007.

In this report the various elements of the safety and nutritional assessment
procedure for genetically modified (GM) plant derived food and feed are
discussed, in particular the potential and limitations of animal feeding trials
for the safety and nutritional testing of whole GM food and feed. The general
principles for the risk assessment of GM plants and derived food and feed are
followed, as described in the EFSA guidance document of the EFSA Scientific Panel
on Genetically Modified Organisms. In Section 1 the mandate, scope and general
principles for risk assessment of GM plant derived food and feed are discussed.
Products under consideration are food and feed derived from GM plants, such as
maize, soybeans, oilseed rape and cotton, modified through the introduction of
one or more genes coding for agronomic input traits like herbicide tolerance
and/or insect resistance. Furthermore GM plant derived food and feed, which have
been obtained through extensive genetic modifications targeted at specific
alterations of metabolic pathways leading to improved nutritional and/or health
characteristics, such as rice containing beta-carotene, soybeans with enhanced
oleic acid content, or tomato with increased concentration of flavonoids, are
considered. The safety assessment of GM plants and derived food and feed follows
a comparative approach, i.e. the food and feed are compared with their non-GM
counterparts in order to identify intended and unintended (unexpected)
differences which subsequently are assessed with respect to their potential
impact on the environment, safety for humans and animals, and nutritional
quality. Key elements of the assessment procedure are the molecular,
compositional, phenotypic and agronomic analysis in order to identify
similarities and differences between the GM plant and its near isogenic
counterpart. The safety assessment is focussed on (i) the presence and
characteristics of newly expressed proteins and other new constituents and
possible changes in the level of natural constituents beyond normal variation,
and on the characteristics of the GM food and feed, and (ii) the possible
occurrence of unintended (unexpected) effects in GM plants due to genetic
modification. In order to identify these effects a comparative phenotypic and
molecular analysis of the GM plant and its near isogenic counterpart is carried
out, in parallel with a targeted analysis of single specific compounds, which
represent important metabolic pathways in the plant like macro and micro
nutrients, known anti-nutrients and toxins. Significant differences may be
indicative of the occurrence of unintended effects, which require further
investigation. Section 2 provides an overview of studies performed for the safety
and nutritional assessment of whole food and feed. Extensive experience has been
built up in recent decades from the safety and nutritional testing in animals of
irradiated foods, novel foods and fruit and vegetables. These approaches are also
relevant for the safety and nutritional testing of whole GM food and feed. Many
feeding trials have been reported in which GM foods like maize, potatoes, rice,
soybeans and tomatoes have been fed to rats or mice for prolonged periods, and
parameters such as body weight, feed consumption, blood chemistry, organ weights,
histopathology etc have been measured. The food and feed under investigation were
derived from GM plants with improved agronomic characteristics like herbicide
tolerance and/or insect resistance. The majority of these experiments did not
indicate clinical effects or histopathological abnormalities in organs or tissues
of exposed animals. In some cases adverse effects were noted, which were
difficult to interpret due to shortcomings in the studies. Many studies have also
been carried out with feed derived from GM plants with agronomic input traits in
target animal species to assess the nutritive value of the feed and their
performance potential. Studies in sheep, pigs, broilers, lactating dairy cows,
and fish, comparing the in vivo bioavailability of nutrients from a range of GM
plants with their near isogenic counterpart and commercial varieties, showed that
they were comparable with those for near isogenic non-GM lines and commercial
varieties. In Section 3 toxicological in vivo, in silico, and in vitro test
methods are discussed which may be applied for the safety and nutritional
assessment of specific compounds present in food and feed or of whole food and
feed derived from GM plants. Moreover the purpose, potential and limitations of
the 90-day rodent feeding trial for the safety and nutritional testing of whole
food and feed have been examined. Methods for single and repeated dose toxicity
testing, reproductive and developmental toxicity testing and immunotoxicity
testing, as described in OECD guideline tests for single well-defined chemicals
are discussed and considered to be adequate for the safety testing of single
substances including new products in GM food and feed. Various in silico and in
vitro methods may contribute to the safety assessment of GM plant derived food
and feed and components thereof, like (i) in silico searches for sequence
homology and/or structural similarity of novel proteins or their degradation
products to known toxic or allergenic proteins, (ii) simulated gastric and
intestinal fluids in order to study the digestive stability of newly expressed
proteins and in vitro systems for analysis of the stability of the novel protein
under heat or other processing conditions, and (iii) in vitro genotoxicity test
methods that screen for point mutations, chromosomal aberrations and DNA
damage/repair. The current performance of the safety assessment of whole foods is
mainly based on the protocols for low-molecular-weight chemicals such as
pharmaceuticals, industrial chemicals, pesticides, food additives and
contaminants. However without adaptation, these protocols have limitations for
testing of whole food and feed. This primarily results from the fact that defined
single substances can be dosed to laboratory animals at very large multiples of
the expected human exposure, thus giving a large margin of safety. In contrast
foodstuffs are bulky, lead to satiation and can only be included in the diet at
much lower multiples of expected human intakes. When testing whole foods, the
possible highest concentration of the GM food and feed in the laboratory animal
diet may be limited because of nutritional imbalance of the diet, or by the
presence of compounds with a known toxicological profile. The aim of the 90-days
rodent feeding study with the whole GM food and feed is to assess potential
unintended effects of toxicological and/or nutritional relevance and to establish
whether the GM food and feed is as safe and nutritious as its traditional
comparator rather than determining qualitative and quantitative intrinsic
toxicity of defined food constituents. The design of the study should be adapted
from the OECD 90-day rodent toxicity study. The precise study design has to take
into account the nature of the food and feed and the characteristics of the new
trait(s) and their intended role in the GM food and feed. A 90-day animal feeding
trial has a large capacity (sensitivity and specificity) to detect potential
toxicological effects of single well defined compounds. This can be concluded
from data reported on the toxicology of a wide range of industrial chemicals,
pharmaceuticals, food substances, environmental, and agricultural chemicals. It
is possible to model the sensitivity of the rat subchronic feeding study for the
detection of hypothetically increased amount of compounds such as anti-nutrients,
toxicants or secondary metabolites. With respect to the detection of potential
unintended effects in whole GM food and feed, it is unlikely that substances
present in small amounts and with a low toxic potential will result in any
observable (unintended) effects in a 90-day rodent feeding study, as they would
be below the no-observed-effect-level and thus of unlikely impact to human health
at normal intake levels. Laboratory animal feeding studies of 90-days duration
appear to be sufficient to pick up adverse effects of diverse compounds that
would also give adverse effects after chronic exposure. This conclusion is based
on literature data from studies investigating whether toxicological effects are
adequately identified in 3-month subchronic studies in rodents, by comparing
findings at 3 and 24 months for a range of different chemicals. The 90-day rodent
feeding study is not designed to detect effects on reproduction or development
other than effects on adult reproductive organ weights and histopathology.
Analyses of available data indicate that, for a wide range of substances,
reproductive and developmental effects are not potentially more sensitive
endpoints than those examined in subchronic toxicity tests. Should there be
structural alerts for reproductive/developmental effects or other indications
from data available on a GM food and feed, then these tests should be considered.
By relating the estimated daily intake, or theoretical maximum daily intake per
capita for a given whole food (or the sum of its individual commercial
constituents) to that consumed on average per rat per day in the subchronic
90-day feeding study, it is possible to establish the margin of exposure (safety
margin) for consumers. Results obtained from testing GM food and feed in rodents
indicate that large (at least 100-fold) 'safety' margins exist between animal
exposure levels without observed adverse effects and estimated human daily
intake. Results of feeding studies with feed derived from GM plants with improved
agronomic properties, carried out in a wide range of livestock species, are
discussed. The studies did not show any biologically relevant differences in the
parameters tested between control and test animals. The studies have shown that
targeted compositional analysis is the cornerstone for the safety assessment of
GM plants modified for agronomic input traits, and once compositional equivalence
has been established, feeding studies with livestock species add little to their
safety assessment. Examples of models for livestock feeding studies with GM
plants with increased concentration of desirable nutrients are provided. Such
studies should be conducted on a case-by-case basis to establish the nutritional
benefits. Possible effects of the new feed resource on animal performance, animal
health, efficacy, and acceptability of the new feed ingredient should be
investigated, and time spans for such studies should be determined on a
case-by-case basis. The feasibility and limitations of human studies with foods
derived from GM plants are discussed, as well as the potential and limitations of
post-market monitoring to detect unintended effects of these foods. Post-market
monitoring is not a substitute for a thorough pre-market risk assessment. In
Section 4 standards for test sample preparation, test materials, diet formulation
and analysis are evaluated. Specific attention is paid to the choice of control
diets and comparators, dietary stability, and nutritional balancing of diets.
When testing whole foods, it is desirable to obtain the highest concentration
possible of the GM food and feed in the laboratory animal diet without causing
nutritional imbalance. Normal practice is to use a minimum of two test dose
levels and negative control with which to create nutritionally equivalent
balanced diets in a comparative protocol. It is recommended to include a relevant
number of commercial varieties as control diets to demonstrate the biological
range of the parameters which are measured in order to assess the biological
relevance of statistically significant differences between the GM plant and its
counterpart. The choice of the comparator for GM food and feed testing is
crucial, and can be found in the parental (near isogenic) line. For modified
macronutrients a comparator is the unmodified form of the macronutrient. For
investigating GM food and feed with enhanced nutritional properties, choices for
control diets should be made on a case-by-case basis. Section 5 provides
information on the collection, analysis and interpretation of data and findings
obtained from animal feeding studies. Data generation for the prediction of
safety and nutritional value of GM plant derived food and feed must be of high
quality in order to perform a proper hazard identification and risk assessment.
This should be based on the use of standardised study designs conducted to the
principles of Good Laboratory Practise, incorporating random quality assurance
audits of all phases of the study. Expert data evaluation and analysis are
critical for establishing any association between exposure and outcome. This
involves specialists from a broad range of scientific disciplines such as
toxicologists, haematologists, clinical biochemists, pathologists, human and
animal nutritionists and also biostatisticians. One of the pivotal requirements
in data analysis is to distinguish those effects which are potentially treatment
related from spurious occurrences or the result of normal individual biological
variation. If differences exist between test and control, comparison to
historical control data from the same laboratory as well as published data for
the strain, sex and age of the animal being investigated is helpful, as well as
data obtained with commercial reference lines. In Section 6 strategies are
outlined for the safety and nutritional assessment of GM plant derived food and
feed. The generation of studies for pre-market assessment of the safety and
nutritional properties of food and feed from GM plants should follow a structured
approach with stepwise development and consideration of the data obtained at each
step in order to formulate the questions to be asked and answered at the next
step (see Fig. 3). Hazards related to the intended genetic modifications are
evaluated applying in silico, in vitro and in vivo safety studies of newly
expressed protein(s), newly formed metabolites, and of natural substances whose
levels may have been altered as a result of gene insertion. Guidelines have been
developed by OECD describing detailed protocols for the safety testing of these
substances in food and feed. A detailed testing strategy should be designed based
on the prior knowledge regarding the biology of these products, so that the
relevant endpoints are measured in the individual test. Testing of the safety and
nutritional value of the whole GM plant or derived food and feed should be
considered where the molecular, compositional, phenotypic, agronomic and other
analyses have demonstrated differences between the GM plant derived food and feed
and their conventional counterpart, apart from the inserted trait(s), or if there
are any indications or remaining uncertainties for the potential occurrence of
unintended effects. In such a case, the testing program should include at least a
90-day rodent feeding study. In the context of the safety and nutritional
assessment of GM plant derived food and feed, the adapted 90-day rodent feeding
study, if triggered by the outcome of the molecular, compositional, phenotypic or
agronomic analysis, functions as a sentinel study designed to assess potential
unintended effects of toxicological and/or nutritional relevance rather than
determining qualitative and quantitative intrinsic toxicity of defined food
constituents. In the situation where molecular, compositional, phenotypic,
agronomic and other analyses have demonstrated equivalence between the GM plant
derived food and feed and their near isogenic counterpart, except for the
inserted trait(s), and do not indicate the occurrence of unintended effects,
experiences with GM plants modified for agronomic input traits have demonstrated
that the performance of 90-day feeding trials with rodents or feeding trials with
target animal species have provided little if anything to the overall safety
assessment (except for added confirmation of safety). The use of 90-days studies
in rodents should be considered for the detection of possible unintended effects
in food and feed derived from GM plants which have been more extensively modified
in order to cope with environmental stress conditions like drought or high salt
conditions, or GM plants with quality or output traits with the purpose to
improve human or animal nutrition and/or health. Ninety-day studies with rodents
are normally of sufficient duration for the identification of general
toxicological effects of compounds that would also give adverse effects after
chronic exposure. In general, long term, chronic toxicity testing of whole GM
food and feed is not expected to generate information additional to what is
already known from in silico/in vitro testing and from subchronic testing. In
cases where structural alerts or other information is available about the
possibly altered occurrence of food components in the GM food and feed compared
to its counterpart, the performance of specific toxicological testing, e.g.
chronic, reproductive, etc., should be considered case-by-case, but
preferentially only for the single substance of concern. Livestock feeding
studies with target animal species should be conducted on a case-by-case basis to
establish the nutritional benefits that might be expected from GM plants with
claimed nutritional/health benefits. Possible effects of the new feed resource on
animal performance, animal health, efficacy, and acceptability of the new feed
ingredient should be investigated, and time spans for such studies should be
determined on a case-by-case basis. There is a need for a more uniform approach
to the design and analysis of animal feeding trials, and in particular for
appropriate statistical analysis of data. The process of data interpretation
requires extensive professional experience of the field, together with a thorough
understanding of the concept of causality. One of the pivotal requirements is to
distinguish those effects which are potentially treatment related from spurious
occurrences or result from normal individual biological variation. Post-market
monitoring is not a substitute for a thorough pre-market risk assessment, neither
should it be considered as a routine need. Knowledge gained through post-market
monitoring might at best describe only broad patterns of human nutritional
exposure. In general it cannot be relied upon as a technique for monitoring
adverse events or other health outcomes related to the consumption of GM plant
derived food and feed. It can be anticipated that in the future the predictive
value of a 90-day rodent feeding studies used for the safety assessment of whole
food and feed will be enhanced by the integration of new technologies like
transcriptomics, proteomics and metabolomics into the experimental risk
assessment approach. Moreover, the use of 'profiling' technologies may also
facilitate a non-targeted approach in compositional analysis in order to aid the
detection of unintended effects in GM plant derived food and feed due to the
genetic modification. These technologies are still under development, and need
validation before they can be used for routine safety assessment purposes. In
Section 7 conclusions and recommendations are presented on:

PMID: 18328408 [PubMed - as supplied by publisher]

46: Plant Cell Rep. 2008 Mar 8. [Epub ahead of print]

Agrobacterium tumefaciens-mediated transformation of poinsettia, Euphorbia
pulcherrima, with virus-derived hairpin RNA constructs confers resistance to
Poinsettia mosaic virus.

Clarke JL, Spetz C, Haugslien S, Xing S, Dees MW, Moe R, Blystad DR.

Plant Health and Plant Protection Division, Norwegian Institute for Agricultural
and Environmental Research, Hoegskoleveien 7, 1432, Aas, Norway,
jihong.liu-clarke@bioforsk.no.

Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima
Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants
of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain
LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA
silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to
transformation, an efficient somatic embryogenesis system was developed for
poinsettia cv. Millenium in which about 75% of the explants produced somatic
embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines
were generated. An average transformation frequency of 2.1% (range 1.2-3.5%) was
revealed. Stable integration of transgenes into the poinsettia nuclear genome was
confirmed by PCR and Southern blot analysis. Both single- and multiple-copy
transgene integration into the poinsettia genome were found among transformants.
Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV
were detected by double antibody sandwich enzyme-linked immunosorbent assay
(DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that
transgene-derived small interfering (si) RNA molecules were detected among the
poinsettia transformants prior to inoculation. The Agrobacterium-mediated
transformation methodology developed in the current study should facilitate
improvement of this ornamental plant with enhanced disease resistance, quality
improvement and desirable colour alteration. Because poinsettia is a non-food,
non-feed plant and is not propagated through sexual reproduction, this is likely
to be more acceptable even in areas where genetically modified crops are
currently not cultivated.

PMID: 18327592 [PubMed - as supplied by publisher]

47: Food Chem Toxicol. 2008 Jun;46(6):1976-84. Epub 2008 Feb 2.

Impact of Bacillus thuringiensis toxin Cry1Ab on rumen epithelial cells (REC) - A
new in vitro model for safety assessment of recombinant food compounds.

Bondzio A, Stumpff F, Schö¶n J, Martens H, Einspanier R.

Department of Veterinary Biochemistry, Freie Universitö¤t Berlin, Oertzenweg 19b,
14163 Berlin, Germany.

The growing use of genetically modified crops necessitates viable screening
methods for safety evaluation of recombinant feed, particularly for ruminants. A
new sheep rumen epithelial cell culture is introduced as an in vitro cell system
for safety evaluation especially focussing on feed and food compounds. We used
lactate dehydrogenase (LDH) release, WST-1 conversion, ATP content and caspase
3/7 activity to evaluate cytotoxicity of Cry1Ab, one of the newly expressed
Bt-proteins in transgene maize. The results were compared to the effects of
valinomycin, a potassium ionophore known to induce cytotoxic effects on a wide
range of cells. Whereas no toxicity of Cry1Ab was observed in short as well as in
long term experiments, even at non-physiological high concentrations, exposure to
valinomycin induced apoptosis and a significant response of all viability
parameters after a number of hours. The ATP content and the WST-1 conversion
reflecting the energy metabolism of the cells appear to be more sensitive
indicators of valinomycin toxicity than the LDH release, a parameter which
reflects the membrane integrity. This study presents an in vitro model system,
that may be useful as a supplementary tool in toxicity screening before testing
substances on animals in vivo.

PMID: 18325653 [PubMed - in process]

48: BMC Biotechnol. 2008 Mar 6;8:26.

Comparison of different real-time PCR chemistries and their suitability for
detection and quantification of genetically modified organisms.

Buh Gasparic M, Cankar K, Zel J, Gruden K.

Department of Biotechnology and Systems Biology, National Institute of Biology,
Vecna pot 111, SI-1000 Ljubljana, Slovenia. meti.buh.gasparic@nib.si

BACKGROUND: The real-time polymerase chain reaction is currently the method of
choice for quantifying nucleic acids in different DNA based quantification
applications. It is widely used also for detecting and quantifying genetically
modified components in food and feed, predominantly employing TaqMan and SYBR
Green real-time PCR chemistries. In our study four alternative chemistries: Lux,
Plexor, Cycling Probe Technology and LNA were extensively evaluated and compared
using TaqMan chemistry as a reference system. RESULTS: Amplicons were designed on
the maize invertase gene and the 5'-junction of inserted transgene and plant
genomic DNA in MON 810 event. Real-time assays were subsequently compared for
their efficiency in PCR amplification, limits of detection and quantification,
repeatability and accuracy to test the performance of the assays. Additionally,
the specificity of established assays was checked on various transgenic and
non-transgenic plant species. The overall applicability of the designed assays
was evaluated, adding practicability and costs issues to the performance
characteristics. CONCLUSION: Although none of the chemistries significantly
outperformed the others, there are certain characteristics that suggest that LNA
technology is an alternative to TaqMan when designing assays for quantitative
analysis. Because LNA probes are much shorter they might be especially
appropriate when high specificity is required and where the design of a common
TaqMan probe is difficult or even impossible due to sequence characteristics.
Plexor on the other hand might be a method of choice for qualitative analysis
when sensitivity, low cost and simplicity of use prevail.

Publication Types:
Comparative Study
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 18325084 [PubMed - indexed for MEDLINE]

49: Neurodegener Dis. 2008;5(3-4):232-6. Epub 2008 Mar 6.

Human recombinant butyrylcholinesterase purified from the milk of transgenic
goats interacts with beta-amyloid fibrils and suppresses their formation in
vitro.

Podoly E, Bruck T, Diamant S, Melamed-Book N, Weiss A, Huang Y, Livnah O,
Langermann S, Wilgus H, Soreq H.

Alexander Silberman Life Sciences Institute, Hebrew University of Jerusalem,
Givat Ram, Jerusalem, Israel.

BACKGROUND: In Alzheimer's disease (AD), brain butyrylcholinesterase (BChE)
co-localizes with beta-amyloid (Abeta) fibrils. AIMS: In vitro testing of the
significance of this phenomenon to AD progress. METHODS: A thioflavine T (ThT)
fluorogenic assay, photo-induced cross-linking and quantifiable electron
microscopy served to compare the effect on Abeta fibril formation induced by
highly purified recombinant human BChE (rBChE) produced in the milk of transgenic
goats with that of serum-derived human BChE. RESULTS: Both proteins at 1:50 and
1:25 ratios to Abeta dose-dependently prolonged the ThT lag time and reduced the
apparent rate of Abeta fibril formation compared to Abeta alone. Photo-induced
cross-linking tests showed that rBChE prolonged the persistence of amyloid
dimers, trimers and tetramers in solution, whereas Abeta alone facilitated
precipitation of such multimers from solution. Transmission electron microscopy
showed that rBChE at 1:100 to Abeta prevented the formation of larger, over
150-nm-long, Abeta fibrils and reduced fibril branching compared to Abeta alone
as quantified by macro programming of Image Pro Plus software. CONCLUSION: Our
findings demonstrate that rBChE interacts with Abeta fibrils and can attenuate
their formation, extension and branching, suggesting further tests of rBChE, with
unlimited supply and no associated health risks, as a therapeutic agent for
delaying the formation of amyloid toxic oligomers in AD patients. 2008 S. Karger
AG, Basel

Publication Types:
Comparative Study

PMID: 18322399 [PubMed - indexed for MEDLINE]

50: Eur J Nutr. 2008 Mar;47(2):99-103. Epub 2008 Mar 4.

Zeaxanthin is bioavailable from genetically modified zeaxanthin-rich potatoes.

Bub A, Mö¶seneder J, Wenzel G, Rechkemmer G, Briviba K.

Federal Research Centre for Nutrition and Food, Institute of Nutritional
Physiology, Haid-und-Neu-Str. 9, 76131 Karlsruhe, Germany. achim.bub@bfel.de

The carotenoid zeaxanthin accumulates in the human macula lutea and protects
retinal cells from blue light damage. However, zeaxanthin intake from food
sources is low. Increasing zeaxanthin in common foods such as potatoes by
traditional plant breeding or by genetic engineering could contribute to an
increased intake of this carotenoid and, consequently, to a decreased risk of
age-related macular degeneration. Our aim was to investigate whether zeaxanthin
from genetically modified zeaxanthin-rich potatoes is bioavailable in humans.
Three men participated in this randomized, controlled double-blinded, crossover
pilot study. All subjects consumed 1,100 g of mashed potatoes, either genetically
modified (Solanum tuberosum L. var. Baltica GM47/18; 3 mg zeaxanthin) or
wild-type control potatoes (Solanum tuberosum L. var. Baltica; 0.14 mg
zeaxanthin). A second treatment was followed after a 7-day wash-out period. The
concentration of zeaxanthin was significantly increased in chylomicrons after
consumption of genetically modified potatoes and 0.27 mg of the 3 mg zeaxanthin
dose could be detected in chylomicrons. Consumption of control potatoes had no
effect on concentrations of zeaxanthin in chylomicrons. After normalization of
chylomicron zeaxanthin for plasma triacylglycerol, the time course of zeaxanthin
concentrations peaked at 7 h after consumption of genetically modified potatoes.
There were no significant differences in the concentrations of other major potato
carotenoids such as lutein and beta-carotene in chylomicrons after consumption of
genetically modified and wild type control potatoes. Thus, consumption of
zeaxanthin-rich potatoes significantly increases chylomicron zeaxanthin
concentrations suggesting that potentially such potatoes could be used as an
important dietary source of zeaxanthin.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18320254 [PubMed - in process]

51: PLoS Biol. 2008 Mar 4;6(3):e68.

Functional adaptation of a plant receptor-kinase paved the way for the evolution
of intracellular root symbioses with bacteria.

Markmann K, Giczey G, Parniske M.

Genetics, Faculty of Biology, Ludwig Maximilians Universitö¤t, Munich, Germany.

Nitrogen-fixing root nodule symbioses (RNS) occur in two major forms-Actinorhiza
and legume-rhizobium symbiosis-which differ in bacterial partner, intracellular
infection pattern, and morphogenesis. The phylogenetic restriction of nodulation
to eurosid angiosperms indicates a common and recent evolutionary invention, but
the molecular steps involved are still obscure. In legumes, at least seven
genes-including the symbiosis receptor-kinase gene SYMRK-are essential for the
interaction with rhizobia bacteria and for the Arbuscular Mycorrhiza (AM)
symbiosis with phosphate-acquiring fungi, which is widespread in occurrence and
believed to date back to the earliest land plants. We show that SYMRK is also
required for Actinorhiza symbiosis of the cucurbit Datisca glomerata with
actinobacteria of the genus Frankia, revealing a common genetic basis for both
forms of RNS. We found that SYMRK exists in at least three different structural
versions, of which the shorter forms from rice and tomato are sufficient for AM,
but not for functional endosymbiosis with bacteria in the legume Lotus japonicus.
Our data support the idea that SYMRK sequence evolution was involved in the
recruitment of a pre-existing signalling network from AM, paving the way for the
evolution of intracellular root symbioses with nitrogen-fixing bacteria.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18318603 [PubMed - indexed for MEDLINE]

52: Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3727-32. Epub 2008 Mar 3.

Cost-effective production of a vaginal protein microbicide to prevent HIV
transmission.

Ramessar K, Rademacher T, Sack M, Stadlmann J, Platis D, Stiegler G, Labrou N,
Altmann F, Ma J, Stö¶ger E, Capell T, Christou P.

Departament de Producciö¨ Vegetal i Ciö¨ncia Forestal, Universitat de Lleida,
Avenida Alcalde Rovira Roure, 191, Lleida, 25198, Spain.

A series of small-molecule microbicides has been developed for vaginal delivery
to prevent heterosexual HIV transmission, but results from human clinical trials
have been disappointing. Protein-based microbicides, such as HIV-specific
monoclonal antibodies, have been considered as an alternative approach. Despite
their promising safety profile and efficacy, the major drawback of such molecules
is the economy of large-scale production in mammalian cells, the current system
of choice. Here, we show that an alternative biomanufacturing platform is now
available for one of the most promising anti-HIV antibodies (2G12). Our data show
that the HIV-neutralization capability of the antibody is equal to or superior to
that of the same antibody produced in CHO cells. We conclude that this protein
production system may provide a means to achieve microbicide ingredient
manufacture at costs that would allow product introduction and manufacture in the
developing world.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18316741 [PubMed - indexed for MEDLINE]

53: J Immunol Methods. 2008 Apr 20;333(1-2):156-166. Epub 2008 Feb 20.

Hybridoma populations enriched for affinity-matured human IgGs yield
high-affinity antibodies specific for botulinum neurotoxins.

Adekar SP, Jones RM, Elias MD, Al-Saleem FH, Root MJ, Simpson LL, Dessain SK.

Cardeza Foundation for Hematologic Research and Kimmel Cancer Center, Thomas
Jefferson University, 1015 Walnut Street, Philadelphia, PA 19107, United States;
Department of Medicine, Thomas Jefferson University, 1025 Walnut Street,
Philadelphia, PA 19107, United States.

The affinity-matured human antibody repertoire may be ideal as a source for
antibody therapeutics against infectious diseases and bioterror agents. Hybridoma
methods for cloning these antibodies have many potential advantages, including
convenience, high-yield antibody expression, and the ability to capture the
antibodies in their native configurations. However, they have been hindered by
hybridoma instability and limited accessibility of antigen-specific,
class-switched human B-cells. Here, we describe an efficient, three-step method
that uses human peripheral blood B-cells to produce stable hybridoma populations
that are highly-enriched for affinity-matured human IgG antibodies. Peripheral
blood mononuclear cells (PBMCs) are (a) selected for expression of CD27, a marker
of post-germinal center B-cells, (b) cultured in vitro to promote B-cell
proliferation and class-switching, and (c) fused to a genetically modified
myeloma cell line. Using this strategy, we cloned 5 IgG antibodies that bind
botulinum neurotoxins (BoNT), the causes of the food-borne paralytic illness,
botulism, and Category A Select Bioterror agents. Two of these antibodies bind
BoNT with low picomolar affinities. One (30B) is the first high-affinity human
antibody to bind serotype B BoNT, and another (6A) is able to neutralize a lethal
dose of serotype A BoNT in vivo in pre- and post-exposure models. This optimized
hybridoma method will broadly enable access to the native human antibody
repertoire.

PMID: 18313069 [PubMed - as supplied by publisher]

54: Learn Mem. 2008 Feb 28;15(3):133-42. Print 2008.

Visual pattern memory requires foraging function in the central complex of
Drosophila.

Wang Z, Pan Y, Li W, Jiang H, Chatzimanolis L, Chang J, Gong Z, Liu L.

State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics,
Chinese Academy of Sciences, Beijing 100101, China.

The role of the foraging (for) gene, which encodes a cyclic
guanosine-3',5'-monophosphate (cGMP)-dependent protein kinase (PKG), in
food-search behavior in Drosophila has been intensively studied. However, its
functions in other complex behaviors have not been well-characterized. Here, we
show experimentally in Drosophila that the for gene is required in the operant
visual learning paradigm. Visual pattern memory was normal in a natural variant
rover (for(R)) but was impaired in another natural variant sitter (for(S)), which
has a lower PKG level. Memory defects in for(S) flies could be rescued by either
constitutive or adult-limited expression of for in the fan-shaped body.
Interestingly, we showed that such rescue also occurred when for was expressed in
the ellipsoid body. Additionally, expression of for in the fifth layer of the
fan-shaped body restored sufficient memory for the pattern parameter "elevation"
but not for "contour orientation," whereas expression of for in the ellipsoid
body restored sufficient memory for both parameters. Our study defines a
Drosophila model for further understanding the role of cGMP-PKG signaling in
associative learning/memory and the neural circuit underlying this for-dependent
visual pattern memory.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18310460 [PubMed - indexed for MEDLINE]

55: Science. 2008 Feb 29;319(5867):1172.

Annette Schavan interview. German science takes an international view. Interview
by Gretchen Vogel.

Schavan A.

Publication Types:
Interview
News

PMID: 18309054 [PubMed - indexed for MEDLINE]

56: Nat Genet. 2008 Mar;40(3):269-70.

Comment on:
Nat Genet. 2008 Mar;40(3):367-72.

Plant breeders go back to nature.

Zamir D.

Publication Types:
Comment
News

PMID: 18305476 [PubMed - indexed for MEDLINE]

57: J Agric Food Chem. 2008 Mar 26;56(6):1818-28. Epub 2008 Feb 28.

Development of a real-time PCR method for the differential detection and
quantification of four solanaceae in GMO analysis: potato (Solanum tuberosum),
tomato (Solanum lycopersicum), eggplant (Solanum melongena), and pepper (Capsicum
annuum).

Chaouachi M, El Malki R, Berard A, Romaniuk M, Laval V, Brunel D, Bertheau Y.

Unitö© Etude du Polymorphisme des Gö©nomes Vö©gö©taux (EPGV) UR1279, Centre National
de Gö©notypage (CNG), 2 rue Gaston Crö©mieux 91057, CP5721, Evry cedex, France.

The labeling of products containing genetically modified organisms (GMO) is
linked to their quantification since a threshold for the presence of fortuitous
GMOs in food has been established. This threshold is calculated from a
combination of two absolute quantification values: one for the specific GMO
target and the second for an endogenous reference gene specific to the taxon.
Thus, the development of reliable methods to quantify GMOs using endogenous
reference genes in complex matrixes such as food and feed is needed. Plant
identification can be difficult in the case of closely related taxa, which
moreover are subject to introgression events. Based on the homology of
beta-fructosidase sequences obtained from public databases, two couples of
consensus primers were designed for the detection, quantification, and
differentiation of four Solanaceae: potato (Solanum tuberosum), tomato (Solanum
lycopersicum), pepper (Capsicum annuum), and eggplant (Solanum melongena).
Sequence variability was studied first using lines and cultivars (intraspecies
sequence variability), then using taxa involved in gene introgressions, and
finally, using taxonomically close taxa (interspecies sequence variability). This
study allowed us to design four highly specific TaqMan-MGB probes. A duplex real
time PCR assay was developed for simultaneous quantification of tomato and
potato. For eggplant and pepper, only simplex real time PCR tests were developed.
The results demonstrated the high specificity and sensitivity of the assays. We
therefore conclude that beta-fructosidase can be used as an endogenous reference
gene for GMO analysis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18303841 [PubMed - in process]

58: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3640-5. Epub 2008 Feb 26.

Microarray analyses reveal that plant mutagenesis may induce more transcriptomic
changes than transgene insertion.

Batista R, Saibo N, Lourenö§o T, Oliveira MM.

Instituto Nacional de Saöºde Dr. Ricardo Jorge, Avenida Padre Cruz, 1649-016
Lisbon, Portugal. rita.batista@insa.min-saude.pt

Controversy regarding genetically modified (GM) plants and their potential impact
on human health contrasts with the tacit acceptance of other plants that were
also modified, but not considered as GM products (e.g., varieties raised through
conventional breeding such as mutagenesis). What is beyond the phenotype of these
improved plants? Should mutagenized plants be treated differently from
transgenics? We have evaluated the extent of transcriptome modification occurring
during rice improvement through transgenesis versus mutation breeding. We used
oligonucleotide microarrays to analyze gene expression in four different pools of
four types of rice plants and respective controls: (i) a gamma-irradiated stable
mutant, (ii) the M1 generation of a 100-Gy gamma-irradiated plant, (iii) a stable
transgenic plant obtained for production of an anticancer antibody, and (iv) the
T1 generation of a transgenic plant produced aiming for abiotic stress
improvement, and all of the unmodified original genotypes as controls. We found
that the improvement of a plant variety through the acquisition of a new desired
trait, using either mutagenesis or transgenesis, may cause stress and thus lead
to an altered expression of untargeted genes. In all of the cases studied, the
observed alteration was more extensive in mutagenized than in transgenic plants.
We propose that the safety assessment of improved plant varieties should be
carried out on a case-by-case basis and not simply restricted to foods obtained
through genetic engineering.

Publication Types:
Comparative Study

PMID: 18303117 [PubMed - indexed for MEDLINE]

59: Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3957-62. Epub 2008 Feb 21.

Antibiotic-resistant soil bacteria in transgenic plant fields.

Demanö¨che S, Sanguin H, Potö© J, Navarro E, Bernillon D, Mavingui P, Wildi W,
Vogel TM, Simonet P.

Universitö© de Lyon, Unitö© Mixte de Recherche 5557, Ecologie Microbienne, Centre
National de la Recherche Scientifique, 69622 Villeurbanne, France.

Understanding the prevalence and polymorphism of antibiotic resistance genes in
soil bacteria and their potential to be transferred horizontally is required to
evaluate the likelihood and ecological (and possibly clinical) consequences of
the transfer of these genes from transgenic plants to soil bacteria. In this
study, we combined culture-dependent and -independent approaches to study the
prevalence and diversity of bla genes in soil bacteria and the potential impact
that a 10-successive-year culture of the transgenic Bt176 corn, which has a
blaTEM marker gene, could have had on the soil bacterial community. The bla gene
encoding resistance to ampicillin belongs to the beta-lactam antibiotic family,
which is widely used in medicine but is readily compromised by bacterial
antibiotic resistance. Our results indicate that soil bacteria are naturally
resistant to a broad spectrum of beta-lactam antibiotics, including the third
cephalosporin generation, which has a slightly stronger discriminating effect on
soil isolates than other cephalosporins. These high resistance levels for a wide
range of antibiotics are partly due to the polymorphism of bla genes, which occur
frequently among soil bacteria. The blaTEM116 gene of the transgenic corn Bt176
investigated here is among those frequently found, thus reducing any risk of
introducing a new bacterial resistance trait from the transgenic material. In
addition, no significant differences were observed in bacterial
antibiotic-resistance levels between transgenic and nontransgenic corn fields,
although the bacterial populations were different.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18292221 [PubMed - indexed for MEDLINE]

60: Rev Panam Salud Publica. 2008 Jan;23(1):52-8.

[Academic production on food labeling in Brazil]

[Article in Portuguese]

Cö¢mara MC, Marinho CL, Guilam MC, Braga AM.

Fundaö§ö¸o Oswaldo Cruz (Fiocruz), Centro de Estudos da Saöºde do Trabalhador e
Ecologia Humana (CESTEH), Brazil. maria.clara@ensp.fiocruz.br

OBJECTIVE: To review and discuss academic production (theses and dissertations)
on the topic of labeling of prepackaged foods in Brazil. METHOD: A search of the
database maintained by the Coordination for the Development of Higher Education
Professionals (CAPES), one of the two Brazilian government research funding and
support agencies, was conducted on the following keywords: "rotulagem"
(labeling), "rotulagem nutricional" (food labeling) and "rö¨tulo de alimentos"
(food labels). The search covered the years 1987 (earliest year available) to
2004. RESULTS: We identified 49 studies on this topic. Content analysis
identified three major themes: the extent to which food labels meet specific
legal requirements (57.2%); the degree to which consumers understand the
information on labels (22.4%); and the labeling of transgenic or
genetically-modified foods (20.4%). CONCLUSIONS: Food labeling is a frequent
topic and is adequately covered by the Brazilian academic production. In most of
the studies, ineffective law enforcement appears to be the main factor in the
lack of compliance with and disrespect for the food labeling rules and
regulations in Brazil.

Publication Types:
English Abstract
Review

PMID: 18291073 [PubMed - indexed for MEDLINE]

61: Lit Med. 2007 Spring;26(1):25-54.

Flora, not fauna: GM culture and agriculture.

McHugh S.

University of New England, USA.

PMID: 18290361 [PubMed - indexed for MEDLINE]

62: Environ Entomol. 2007 Oct;36(5):1275-82.

Degradation of Cry1Ac protein within transgenic Bacillus thuringiensis rice
tissues under field and laboratory conditions.

Li Y, Wu K, Zhang Y, Yuan G.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of
Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094,
China.

To clarify the environmental fate of the Cry1Ac protein from Bacillus
thuringiensis subsp. kurstaki (Bt) contained in transgenic rice plant stubble
after harvest, degradation was monitored under field conditions using an
enzyme-linked immunosorbent assay. In stalks, Cry1Ac protein concentration
decreased rapidly to 50% of the initial amount during the first month after
harvest; subsequently, the degradation decreased gradually reaching 21.3% when
the experiment was terminated after 7 mo. A similar degradation pattern of the
Cry1Ac protein was observed in rice roots. However, when the temperature
increased in April of the following spring, protein degradation resumed, and no
protein could be detected by the end of the experiment. In addition, a laboratory
experiment was conducted to study the persistence of Cry1Ac protein released from
rice tissue in water and paddy soil. The protein released from leaves degraded
rapidly in paddy soil under flooded conditions during the first 20 d and
plateaued until the termination of this trial at 135 d, when 15.3% of the initial
amount was still detectable. In water, the Cry1Ac protein degraded more slowly
than in soil but never entered a relatively stable phase as in soil. The
degradation rate of Cry1Ac protein was significantly faster in nonsterile water
than in sterile water. These results indicate that the soil environment can
increase the degradation of Bt protein contained in plant residues. Therefore,
plowing a field immediately after harvest could be an effective method for
decreasing the persistence of Bt protein in transgenic rice fields.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18284753 [PubMed - indexed for MEDLINE]

63: Environ Entomol. 2007 Oct;36(5):1269-74.

Response of ground beetle (Coleoptera: Carabidae) field populations to four years
of Lepidoptera-specific Bt corn production.

Floate KD, Cö¡rcamo HA, Blackshaw RE, Postman B, Bourassa S.

Agriculture and Agri-Food Canada, Lethbridge Research Centre, 5403 1st Ave. S.,
Lethbridge, AB, Canada T1J 4B1. floatek@agr.gc.ca

Pitfall traps were used to monitor populations of ground beetles (Coleoptera:
Carabidae) in plots of corn grown in continuous cultivation during a 4-yr period
(2000-2003). Treatments included transgenic corn expressing a Bt Cry protein with
efficacy specific against Lepidoptera (Bt), conventional corn grown with
insecticide application (I), and the same conventional cultivar grown without
insecticide application (NI). Mixed-model analyses of variance were performed on
pitfall captures of beetles combined across weeks to give seasonal sums. Effects
of corn treatment were not detected (P > 0.05) on total beetle abundance or
species richness in any year. Effects of corn treatment on individual taxa were
detected (P < 0.05) for 3 of the 39 species-by-year combinations examined.
Effects of near significance (P < 0.08) were detected for an additional two
species. In 2001, captures of Amara farcta Leconte and Harpalus amputatus Say
were lower in Bt plots than in I or NI plots. In 2003, captures of Amara
apricaria (Paykull) and Amara carinata (Leconte) were higher in Bt plots than in
I or NI plots. Also in 2003, captures of Poecilus scitulus Leconte were higher in
I plots than in Bt or NI plots. These patterns were not repeated among years.
Results of this study indicate that cultivation of Lepidoptera-specific Bt corn
in southern Alberta does not appreciably affect ground beetle populations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18284752 [PubMed - indexed for MEDLINE]

64: Environ Entomol. 2007 Oct;36(5):1254-68.

Effects of insecticide-treated and Lepidopteran-active Bt transgenic sweet corn
on the abundance and diversity of arthropods.

Rose R, Dively GP.

Biotechnology Regulatory Services, USDA, Animal and Plant Health Inspection
Service, Unit 147, 4700 River Rd., Riverdale, MD 20737, USA.

A field study was conducted over 2 yr to determine the effects of transgenic
sweet corn containing a gene from the bacterium Bacillus thuringiensis (Bt) on
the diversity and abundance of nontarget arthropods. The Bt hybrid (expressing
Cry1Ab endotoxin for lepidopteran control) was compared with near-isogenic non-Bt
and Bt hybrids treated with a foliar insecticide and with a near-isogenic non-Bt
hybrid without insecticides. Plant inspections, sticky cards, and pitfall traps
were used to sample a total of 573,672 arthropods, representing 128 taxonomic
groups in 95 families and 18 orders. Overall biodiversity and community-level
responses were not significantly affected by the transgenic hybrid. The Bt hybrid
also had no significant adverse effects on population densities of specific
nontarget herbivores, decomposers, and natural enemies enumerated at the family
level during the crop cycle. As expected, the insecticide lambda-cyhalothrin had
broad negative impacts on the abundance of many nontarget arthropods. One
insecticide application in the Bt plots reduced the overall abundance of the
natural enemy community by 21-48%. Five applications in the non-Bt plots reduced
natural enemy communities by 33-70%. Nontarget communities affected in the
insecticide-treated Bt plots exhibited some recovery, but communities exposed to
five applications showed no trends toward recovery during the crop cycle. This
study clearly showed that the nontarget effects of Bt transgenic sweet corn on
natural enemies and other arthropods were minimal and far less than the
community-level disruptions caused by lambda-cyhalothrin.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18284751 [PubMed - indexed for MEDLINE]

65: Poult Sci. 2008 Mar;87(3):475-9.

Performance of laying hens fed diets containing DAS-59122-7 maize grain compared
with diets containing nontransgenic maize grain.

Jacobs CM, Utterback PL, Parsons CM, Rice D, Smith B, Hinds M, Liebergesell M,
Sauber T.

Department of Animal Sciences, University of Illinois, Urbana 61801, USA.

An experiment using 216 Hy-Line W-36 pullets was conducted to evaluate transgenic
maize grain containing the cry34Ab1 and cry35Ab1 genes from a Bacillus
thuringiensis (Bt) strain and the phosphinothricin ace-tyltransferase (pat) gene
from Streptomyces viridochromogenes. Expression of the cry34Ab1 and cry35Ab1
genes confers resistance to corn rootworms, and the pat gene confers tolerance to
herbicides containing glufosinate-ammonium. Pullets (20 wk of age) were placed in
cage lots (3 hens/cage, 2 cages/lot) and were randomly assigned to 1 of 3
corn-soybean meal dietary treatments (12 lots/treatment) formulated with the
following maize grains: near-isogenic control (control), conventional maize, and
transgenic test corn line 59122 containing event DAS-59122-7. Differences between
59122 and control group means were evaluated with statistical significance at P <
0.05. Body weight and gain, egg production, egg mass, and feed efficiency for
hens fed the 59122 corn were not significantly different from the respective
values for hens fed diets formulated with control maize grain. Egg component
weights, Haugh unit measures, and egg weight class distribution were similar
regardless of the corn source. This research indicates that performance of hens
fed diets containing 59122 maize grain, as measured by egg production and egg
quality, was similar to that of hens fed diets formulated with near-isogenic corn
grain.

PMID: 18281573 [PubMed - indexed for MEDLINE]

66: Nat Genet. 2008 Mar;40(3):367-72. Epub 2008 Feb 17.

Comment in:
Nat Genet. 2008 Mar;40(3):269-70.

A phenylalanine in DGAT is a key determinant of oil content and composition in
maize.

Zheng P, Allen WB, Roesler K, Williams ME, Zhang S, Li J, Glassman K, Ranch J,
Nubel D, Solawetz W, Bhattramakki D, Llaca V, Deschamps S, Zhong GY, Tarczynski
MC, Shen B.

Pioneer Hi-Bred International Inc., A DuPont Company, 7300 NW 62nd Avenue, PO Box
1004, Johnston, Iowa 50131, USA.

Plant oil is an important renewable resource for biodiesel production and for
dietary consumption by humans and livestock. Through genetic mapping of the oil
trait in plants, studies have reported multiple quantitative trait loci (QTLs)
with small effects, but the molecular basis of oil QTLs remains largely unknown.
Here we show that a high-oil QTL (qHO6) affecting maize seed oil and oleic-acid
contents encodes an acyl-CoA:diacylglycerol acyltransferase (DGAT1-2), which
catalyzes the final step of oil synthesis. We further show that a phenylalanine
insertion in DGAT1-2 at position 469 (F469) is responsible for the increased oil
and oleic-acid contents. The DGAT1-2 allele with F469 is ancestral, whereas the
allele without F469 is a more recent mutant selected by domestication or
breeding. Ectopic expression of the high-oil DGAT1-2 allele increases oil and
oleic-acid contents by up to 41% and 107%, respectively. This work provides
insights into the molecular basis of natural variation of oil and oleic-acid
contents in plants and highlights DGAT as a promising target for increasing oil
and oleic-acid contents in other crops.

PMID: 18278045 [PubMed - indexed for MEDLINE]

67: Physiol Plant. 2008 Mar;132(3):384-96.

Overexpression of zeaxanthin epoxidase gene enhances the sensitivity of tomato
PSII photoinhibition to high light and chilling stress.

Wang N, Fang W, Han H, Sui N, Li B, Meng QW.

College of Life Science, State Key Laboratory of Crop Biology, Shandong
Agricultural University, Tai'an 271018, China.

A tomato (Lycopersicon esculentum Mill.) zeaxanthin epoxidase gene (LeZE) was
isolated. The deduced amino acid sequence of LeZE showed high identities with
zeaxanthin epoxidase in other plant species. Northern blot analysis showed that
the mRNA accumulation of LeZE in the wild-type (WT) was not induced by light and
temperature but regulated by the diurnal rhythm. The sense transgenic plants were
obtained under the control of the cauliflower mosaic virus 35S promoter
(35S-CaMV). Northern and western blot analysis confirmed that sense LeZE was
transferred into the tomato genome and overexpressed. The ratio of (A + Z)/(V + A
+ Z) and the values of non-photochemical quenching were lower in transgenic
plants than in WT plants under high light and chilling stress with low
irradiance. The O(2) evolution rate and the maximal photochemical efficiency of
PSII (Fv/Fm) in transgenic plants decreased more quickly during both stresses and
recovered slower than that in WT under optimal conditions. These results
suggested that overexpression of LeZE impaired the function of the xanthophyll
cycle and aggravated PSII photoinhibition in tomato under high light and chilling
stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18275469 [PubMed - indexed for MEDLINE]

68: Dokl Biochem Biophys. 2007 Nov-Dec;417:327-30.

Methylation of GCGG sites of the patatin promoter is organ-specific and inversely
correlates with its activity.

Romanov GA, Naumkina EM, Ashapkin VV, Vanyushin BF.

Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, ul.
Botanicheskaya 35, Moscow 127276, Russia.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18274451 [PubMed - indexed for MEDLINE]

69: J Exp Bot. 2008;59(2):377-87. Epub 2008 Feb 10.

Senescence-induced ectopic expression of the A. tumefaciens ipt gene in wheat
delays leaf senescence, increases cytokinin content, nitrate influx, and nitrate
reductase activity, but does not affect grain yield.

Sykorovö¡ B, Kuresovö¡ G, Daskalova S, Trckovö¡ M, Hoyerovö¡ K, Raimanovö¡ I, Motyka
V, Trö¡vnö­ckovö¡ A, Elliott MC, Kamö­nek M.

Institute of Experimental Botany, Academy of Sciences of the Czech Republic,
Rozvojovö¡ 263, 16502 Prague 6, Czech Republic.

The manipulation of cytokinin levels by senescence-regulated expression of the
Agrobacterium tumefaciens ipt gene through its control by the Arabidopsis SAG12
(senescence-associated gene 12) promoter is an efficient tool for the
prolongation of leaf photosynthetic activity which potentially can affect plant
productivity. In the present study, the efficiency of this approach was tested on
wheat (Triticum aestivum L.)-a monocarpic plant characterized by a fast switch
from vegetative to reproductive growth, and rapid translocation of metabolites
from leaves to developing grains after anthesis. When compared with the wild-type
(WT) control plants, the SAG12::ipt wheat plants exhibited delayed chlorophyll
degradation only when grown under limited nitrogen (N) supply. Ten days after
anthesis the content of chlorophyll and bioactive cytokinins of the first (flag)
leaf of the transgenic plants was 32% and 65% higher, respectively, than that of
the control. There was a progressive increase in nitrate influx and nitrate
reductase activity. However, the SAG12::ipt and the WT plants did not show
differences in yield-related parameters including number of grains and grain
weight. These results suggest that the delay of leaf senescence in wheat also
delays the translocation of metabolites from leaves to developing grains, as
indicated by higher accumulation of ((15)N-labelled) N in spikes of control
compared with transgenic plants prior to anthesis. This delay interferes with the
wheat reproductive strategy that is based on a fast programmed translocation of
metabolites from the senescing leaves to the reproductive sinks shortly after
anthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18267946 [PubMed - indexed for MEDLINE]

70: Nat Biotechnol. 2008 Feb;26(2):161-2.

Action needed to harmonize regulation of low-level presence of biotech traits.

Krueger R, Le Buanec B.

Publication Types:
Letter

PMID: 18259165 [PubMed - indexed for MEDLINE]

71: Nat Biotechnol. 2008 Feb;26(2):159-60.

FDA on transgenic animals--a dog's breakfast?

Miller HI.

Publication Types:
Letter

PMID: 18259164 [PubMed - indexed for MEDLINE]

72: Nat Biotechnol. 2008 Feb;26(2):139-40.

Scientists cry foul as Europe plays politics with GM crops.

Hodgson J.

Publication Types:
News

PMID: 18259156 [PubMed - indexed for MEDLINE]

73: J Zhejiang Univ Sci B. 2008 Feb;9(2):132-40.

Molecular and functional comparisons of the vacuolar Na+/H+ exchangers originated
from glycophytic and halophytic species.

Li JY, He XW, Xu L, Zhou J, Wu P, Shou HX, Zhang FC.

State Key Laboratory of Plant Physiology and Biochemistry, College of Life
Sciences, Zhejiang University, Hangzhou 310058, China.

A novel vacuolar Na+/H+ exchanger, CgNHX1, was cloned from a halophytic species
Chenopodium glaucum by using reverse transcriptase-polymerase chain reaction
(RT-PCR) and rapid amplification of cDNA ends (RACE) technique. Sequence
alignment and phylogenetic analysis of 22 NHX genes from GenBank as well as the
new CgNHX1 gene indicate that NHX genes shared a great degree of similarity,
regardless of their glycophytic or halophytic origin. Expression of the CgNHX1
gene was induced by NaCl and peaked at 400 mmol/L NaCl. Overexpression of NHX1
genes in rice enhanced their tolerance to salt stress. However, there is no
significant difference in salt tolerance among the transgenic rice plants
overexpressing the NHX1 genes from either glycophytic or halophytic species. The
Na+ content of both the wild type (WT) and transgenic plants increased when
exposed to 50 and 100 mmol/L NaCl, and the Na+ concentration in transgenic plants
was marginally higher than that of WT. Our data demonstrate that the
overexpression of the NHX1 gene from either glycophytic or halophytic species
resulted in the enhanced tolerance to salt stress at a similar level, suggesting
that NHX gene per se might not be the reason accounting for the difference in
salt tolerance between glycophytes and halophytes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18257135 [PubMed - indexed for MEDLINE]

74: J Exp Bot. 2008;59(2):213-23. Epub 2008 Feb 5.

Effect of the cauliflower Or transgene on carotenoid accumulation and chromoplast
formation in transgenic potato tubers.

Lopez AB, Van Eck J, Conlin BJ, Paolillo DJ, O'Neill J, Li L.

USDA-ARS, Plant, Soil and Nutrition Laboratory, Cornell University, Ithaca, NY
14853, USA.

Transgenic plants have facilitated our understanding of the functional roles of
genes and the metabolic processes affected in plants. Recently, the Or gene was
isolated from an orange cauliflower mutant and it was shown that the Or gene
could serve as a novel genetic tool to enrich carotenoid content in transgenic
potato tubers. An in-depth characterization of these Or transgenic lines is
presented here. It was found that the Or transgene may facilitate the
identification of potential rate-limiting step(s) of the carotenoid biosynthetic
pathway. The Or transgenic tubers accumulated not only increased levels of
carotenoids that normally are present in the controls, but also three additional
metabolite intermediates of phytoene, phytofluene, and zeta-carotene, indicating
that the desaturation steps became limiting following the expression of the Or
transgene. Moreover, we observed that long-term cold storage greatly enhanced
carotenoid content in the Or transgenic tubers to a level of 10-fold over
controls. Expression of the Or transgene in the transgenic plants caused no
dramatic changes in the transcript levels of the endogenous carotenoid
biosynthetic genes, which is in agreement with the Or gene not directly
controlling carotenoid biosynthesis. Microscope analysis revealed that the Or
transgene conferred the formation of chromoplasts containing carotenoid
sequestering structures in a heterologous system. Such structures were not
observed in tubers of potato cultivars that accumulate high levels of
carotenoids. Collectively, these results provide direct evidence demonstrating
that the Or gene indeed controls chromoplast differentiation and that regulation
of chromoplast formation can have a profound effect on carotenoid accumulation in
plants.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18256051 [PubMed - indexed for MEDLINE]

75: Food Chem Toxicol. 2007 Dec 25. [Epub ahead of print]

Identification of potentially emerging food safety issues by analysis of reports
published by the European Community's Rapid Alert System for Food and Feed
(RASFF) during a four-year period.

Kleter GA, Prandini A, Filippi L, Marvin HJ.

RIKILT Á€“ Institute of Food Safety, Wageningen University and Research Center,
P.O. Box 230, NL-6700 AE Wageningen, The Netherlands.

The SAFE FOODS project undertakes to design a new approach towards the early
identification of emerging food safety hazards. This study explored the utility
of notifications filed through RASFF, the European Commission's Rapid Alert
System for Food and Feed, to identify emerging trends in food safety issues.
RASFF information and alert notifications published in the four-year period of
July 2003-June 2007 were assigned to categories of products and hazards. For
chronological trend analysis, a basic time unit of three months was chosen. Data
within each hazard category were analyzed for chronological trends, relationships
between product and hazard categories, regions of origin, and countries filing
the notifications. Conspicuous trends that were observed included a rise in the
incidence of food contact substances, particularly 2-isopropyl-thioxanthone, as
well as of chemical substances migrating from utensils and fraud-related issues.
Temporary increases were noted in the incidences of the unauthorized dye Para
Red, genetically modified organisms, the pesticide isophenfos-methyl, and herring
worm, Anisakis simplex. National and European authorities themselves have
signaled these conspicuous trends and taken measures. It is recommended to add
complementary data to RASFF data, including safety assessments, risk management
measures, background data on hazards and surveillance patterns, for a holistic
approach towards early identification of emerging hazards.

PMID: 18255210 [PubMed - as supplied by publisher]

76: J Agric Food Chem. 2008 Mar 12;56(5):1589-94. Epub 2008 Feb 7.

Starch physicochemical properties and their associations with microsatellite
alleles of starch-synthesizing genes in a rice RIL population.

Bao J, Jin L, Xiao P, Shen S, Sun M, Corke H.

Institute of Nuclear-Agricultural Sciences, Key Laboratory of Chinese Ministry of
Agriculture for Nuclear-Agricultural Sciences, College of Agriculture and
Biotechnology, Zhejiang University, Hangzhou, People's Republic of China.
jsbao@zju.edu.cn

The physicochemical properties of starch, such as apparent amylose content,
gelatinization temperature, and pasting viscosities, determine the eating,
cooking, and processing qualities of various products of rice. A recombinant
inbred line (RIL) population derived from the reciprocal cross of Lemont (a
premium high-quality tropical japonica rice) and Jiayu 293 (a high-yield but
low-quality indica rice) was used to test the association of microsatellite
markers of starch-synthesizing genes with starch quality parameters. The results
confirmed the association of Wx and starch synthase I (SSI) alleles with various
starch properties measured in rice flour. However, the starch properties were not
associated with the starch branching enzyme 1 (SBE 1) gene alleles.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18254594 [PubMed - indexed for MEDLINE]

77: J Exp Bot. 2008;59(2):315-25. Epub 2008 Feb 4.

Decreased expression of plastidial adenylate kinase in potato tubers results in
an enhanced rate of respiration and a stimulation of starch synthesis that is
attributable to post-translational redox-activation of ADP-glucose
pyrophosphorylase.

Oliver SN, Tiessen A, Fernie AR, Geigenberger P.

Max-Planck-Institute of Molecular Plant Physiology, Am Muehlenberg 1, D-14476
Potsdam-Golm, Germany.

Adenine nucleotides are of general importance for many aspects of cell function,
but their role in the regulation of biosynthetic processes is still unclear. It
was previously reported that decreased expression of plastidial adenylate kinase,
catalysing the interconversion of ATP and AMP to ADP, leads to increased
adenylate pools and starch content in transgenic potato tubers. However, the
underlying mechanisms were not elucidated. Here, it is shown that decreased
expression of plastidial adenylate kinase in growing tubers leads to increased
rates of respiratory oxygen consumption and increased carbon fluxes into starch.
Increased rates of starch synthesis were accompanied by post-translational
redox-activation of ADP-glucose pyrophosphorylase (AGPase), catalysing the key
regulatory step of starch synthesis in the plastid, while there were no
substantial changes in metabolic intermediates or sugar levels. A similar
increase in post-translational redox-activation of AGPase was found after
supplying adenine to wild-type potato tuber discs to increase adenine nucleotide
levels. Results provide first evidence for a link between redox-activation of
AGPase and adenine nucleotide levels in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18252705 [PubMed - indexed for MEDLINE]

78: Physiol Plant. 2007 Sep;131(1):106-21.

The rice Mybleu transcription factor increases tolerance to oxygen deprivation in
Arabidopsis plants.

Mattana M, Vannini C, Espen L, Bracale M, Genga A, Marsoni M, Iriti M, Bonazza V,
Romagnoli F, Baldoni E, Coraggio I, Locatelli F.

Istituto di Biologia e Biotecnologia Agraria, CNR, via E Bassini 15, 20133
Milano, Italy.

Mybleu is a natural incomplete transcription factor of rice (Oryza sativa),
consisting of a partial Myb repeat followed by a short leucine zipper. We
previously showed its localization to the apical region of rice roots and
coleoptiles. Specifically, in coleoptiles, Mybleu is expressed under both aerobic
and anaerobic conditions, whereas in roots, it is expressed only under aerobic
conditions. Mybleu is able to dimerize with canonical leucine zippers and to
activate transcription selectively. To investigate Mybleu function in vivo, we
transformed Arabidopsis thaliana and evaluated several morphological,
physiological and biochemical parameters. In agreement with a hypothesized role
of Mybleu in cell elongation in the differentiation zone, we found that the
constitutive expression of this transcription factor in Arabidopsis induced
elongation in the primary roots and in the internodal region of the floral stem;
we also observed a modification of the root apex morphology in transformed lines.
Based on the high expression of Mybleu in anaerobic rice coleoptiles, we studied
the role of this transcription factor in transgenic plants grown under low-oxygen
conditions. We found that overexpression of this transcription factor increased
tolerance to oxygen deficit. In transgenic plants, this effect may depend both on
the maintenance of a higher metabolism during stress and on the higher expression
levels of certain genes involved in the anaerobic response.

PMID: 18251929 [PubMed - indexed for MEDLINE]

79: Physiol Plant. 2008 Feb;132(2):236-53.

Transcriptome analyses give insights into selenium-stress responses and selenium
tolerance mechanisms in Arabidopsis.

Van Hoewyk D, Takahashi H, Inoue E, Hess A, Tamaoki M, Pilon-Smits EA.

Department of Biology, Colorado State University, Fort Collins, CO 80523, USA.

Selenate is chemically similar to sulfate and can be taken up and assimilated by
plants via the same transporters and enzymes. In contrast to many other
organisms, selenium (Se) has not been shown to be essential for higher plants. In
excess, Se is toxic and restricts development. Both Se deficiency and toxicity
pose problems worldwide. To obtain better insights into the effects of Se on
plant metabolism and into plant mechanisms involved in Se tolerance, the
transcriptome of Arabidopsis plants grown with or without selenate was studied
and Se-responsive genes identified. Roots and shoots exhibited different
Se-related changes in gene regulation and metabolism. Many genes involved in
sulfur (S) uptake and assimilation were upregulated. Accordingly, Se treatment
enhanced sulfate levels in plants, but the quantity of organic S metabolites
decreased. Transcripts regulating the synthesis and signaling of ethylene and
jasmonic acid were also upregulated by Se. Arabidopsis mutants defective in
ethylene or jasmonate response pathways exhibited reduced tolerance to Se,
suggesting an important role for these two stress hormones in Se tolerance.
Selenate upregulated a variety of transcripts that were also reportedly induced
by salt and osmotic stress. Selenate appeared to repress plant development, as
suggested by the downregulation of genes involved in cell wall synthesis and
auxin-regulated proteins. The Se-responsive genes discovered in this study may
help create plants that can better tolerate and accumulate Se, which may enhance
the effectiveness of Se phytoremediation or serve as Se-fortified food.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18251864 [PubMed - indexed for MEDLINE]

80: Appl Environ Microbiol. 2008 Apr;74(7):2129-34. Epub 2008 Feb 1.

Construction of sterile ime1Delta-transgenic Saccharomyces cerevisiae wine yeasts
unable to disseminate in nature.

Ramö­rez M, Ambrona J.

Departamento de Microbiologö­a (Antiguo Rectorado), Facultad de Ciencias,
Universidad de Extremadura, 06071 Badajoz, Spain. mramirez@unex.es

The use of new transgenic yeasts in industry carries a potential environmental
risk because their dispersal, introducing new artificial genetic combinations
into nature, could have unpredictable consequences. This risk could be avoided by
using sterile transgenic yeasts that are unable to sporulate and mate with wild
yeasts. These sterile yeasts would not survive the annual cyclic harvesting
periods, being condemned to disappear in the wineries and vineyards in less than
a year. We have constructed new ime1Delta wine yeasts that are unable to
sporulate and mate, bear easy-to-detect genetic markers, and quickly disappear in
grape must fermentation immediately after sporulation of the yeast population.
These sterile yeasts maintained the same biotechnological properties as their
parent yeasts without any detectable deleterious effect of the ime1Delta
mutation. These yeasts are therefore interesting biotechnologically for food
industry applications and for genetically modified microorganism environmental
monitoring studies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18245242 [PubMed - in process]

81: Appetite. 2008 Jul;51(1):58-68. Epub 2007 Dec 15.

Knowledge, attitudes towards and acceptability of genetic modification in
Germany.

Christoph IB, Bruhn M, Roosen J.

Federal Research Centre for Nutrition and Food (BfEL), Department of Food
Economics, Hermann-Weigmann-Strasse, 24103 Kiel, Germany.

Genetic modification remains a controversial issue. The aim of this study is to
analyse the attitudes towards genetic modification, the knowledge about it and
its acceptability in different application areas among German consumers. Results
are based on a survey from spring 2005. An exploratory factor analysis is
conducted to identify the attitudes towards genetic modification. The identified
factors are used in a cluster analysis that identified a cluster of supporters,
of opponents and a group of indifferent consumers. Respondents' knowledge of
genetics and biotechnology differs among the found clusters without revealing a
clear relationship between knowledge and support of genetic modification. The
acceptability of genetic modification varies by application area and cluster, and
genetically modified non-food products are more widely accepted than food
products. The perception of personal health risks has high explanatory power for
attitudes and acceptability.

PMID: 18243411 [PubMed - in process]

82: Trends Biotechnol. 2008 Mar;26(3):122-5.

Is biotechnology a victim of anti-science bias in scientific journals?

Miller HI, Morandini P, Ammann K.

The Hoover Institution, 434 Galvez Mall, Stanford University, Stanford, CA
94305-6010, USA. miller@hoover.stanford.edu

Primarily outside the scientific community, misapprehensions and misinformation
about recombinant DNA-modified (also known as 'genetically modified', or 'GM')
plants have generated significant 'pseudo-controversy' over their safety that has
resulted in unscientific and excessive regulation (with attendant inflated
development costs) and disappointing progress. But pseudo-controversy and
sensational claims have originated within the scientific community as well, and
even scholarly journals' treatment of the subject has been at times unscientific,
one-sided and irresponsible. These shortcomings have helped to perpetuate 'The
Big Lie' - that recombinant DNA technology applied to agriculture and food
production is unproven, unsafe, untested, unregulated and unwanted. Those
misconceptions, in turn, have given rise to unwarranted opposition and tortuous,
distorted public policy.

Publication Types:
Review

PMID: 18243381 [PubMed - indexed for MEDLINE]

83: Anal Bioanal Chem. 2008 Feb 2. [Epub ahead of print]

Advances in molecular techniques for the detection and quantification of
genetically modified organisms.

Elenis DS, Kalogianni DP, Glynou K, Ioannou PC, Christopoulos TK.

Department of Chemistry, University of Athens, Athens, 15771, Greece.

Progress in genetic engineering has led to the introduction of genetically
modified organisms (GMOs) whose genomes have been altered by the integration of a
novel sequence conferring a new trait. To allow consumers an informed choice,
many countries require food products to be labeled if the GMO content exceeds a
certain threshold. Consequently, the development of analytical methods for GMO
screening and quantification is of great interest. Exponential amplification by
the polymerase chain reaction (PCR) remains a central step in molecular methods
of GMO detection and quantification. In order to meet the challenge posed by the
continuously increasing number of GMOs, various multiplex assays have been
developed for the simultaneous amplification and/or detection of several GMOs.
Classical agarose gel electrophoresis is being replaced by capillary
electrophoresis (CE) systems, including CE chips, for the rapid and automatable
separation of amplified fragments. Microtiter well-based hybridization assays
allow high-throughput analysis of many samples in a single plate. Microarrays
have been introduced in GMO screening as a technique for the simultaneous
multianalyte detection of amplified sequences. Various types of biosensors,
including surface plasmon resonance sensors, quartz crystal microbalance
piezoelectric sensors, thin-film optical sensors, dry-reagent dipstick-type
sensors and electrochemical sensors were introduced in GMO screening because they
offer simplicity and lower cost. GMO quantification is performed by real-time PCR
(rt-QPCR) and competitive PCR. New endogenous reference genes have been
validated. rt-QPCR is the most widely used approach. Multiplexing is another
trend in this field. Strategies for high-throughput multiplex competitive
quantitative PCR have been reported.

PMID: 18239909 [PubMed - as supplied by publisher]

84: Sci China C Life Sci. 2008 Feb;51(2):145-56.

Leaf surface factors of transgenic Bt cotton associated with the feeding
behaviors of cotton aphids: a case study on non-target effects.

Xue K, Deng S, Wang R, Yan F, Xu C.

College of Life Sciences, Peking University, Beijing 100871, China.

The present paper reports case study results of the risk assessment of transgenic
Bt cotton on a non-target pest, cotton aphid, Aphis gossypii. Several types of
techniques, i.e., electrical penetration graph (EPG), light and electron
microscopy, bioassays and chemical analysis, were applied to investigate physical
and chemical leaf factors of 2 transgenic Bt cotton lines (GK12 and GK19) and
their parental non-Bt cotton line (Simian3) associated with searching and feeding
behaviors of cotton aphids on leaves or leaf extracts of cotton plants. EPG
results showed that there were some differences among behaviors of cotton aphids
on 2 Bt cotton and 1 non-Bt cotton lines. Cotton aphids performed similarly to
leaf surface extracts from 3 cotton lines; and leaf surface chemicals, mainly
volatiles and waxes, were almost identical in the components and concentrations
among the cotton lines. However, three cotton lines were quite different from
each other in the densities of certain kinds of covering trichomes. Therefore,
the relationships between the physical characteristics and the searching
behaviors of cotton aphids on the three cotton lines were constructed as the
regression equations. Glandular trichomes and covering trichomes with 5 branches
influenced the cotton aphids' searching behaviors effectively; and other
trichomes with other branches affected aphids in varying ways. These results
demonstrated that leaf surface physical factors of transgenic Bt cotton lines
different from their parental non-Bt line could affect the penetration behaviors
of non-target cotton aphids. Cotton aphids penetrate and feed more easily on two
Bt cotton lines than on the non-Bt cotton line.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18239893 [PubMed - indexed for MEDLINE]

85: J Econ Entomol. 2007 Dec;100(6):1880-6.

Evaluation of transgenic Bacillus thuringiensis corn hybrids against
Cry1Ab-susceptible and -resistant sugarcane borer (Lepidoptera: Crambidae).

Wu X, Huang F, Leonard BR, Moore SH.

Department of Entomology, Louisiana State University Agricultural Center, Baton
Rouge, LA 70803, USA.

A Louisiana strain of the sugarcane borer, Diatraea saccharalis (F.)
(Lepidoptera: Crambidae), was selected for resistance to the CrylAb protein of
Bacillus thuringiensis (Bt) by using an F2 screening procedure. Survival of
Bt-resistant, -susceptible, and -heterozygous genotypes of sugarcane borer was
evaluated on vegetative and reproductive stages of five non-Bt and seven Bt field
corn, Zea mays L., hybrids in a greenhouse study. Larval survival was recorded 21
d after infestation of neonates on potted plants. Larval survival across the
three sugarcane borer genotypes and five non-Bt corn hybrids after 21 d ranged
from 23.6 +/- 5.2% (mean +/- SEM) to 57.5 +/- 5.2%. Mean survival of
Cry1Ab-resistant larvae on vegetative and reproductive plant stages was 12 and
21%, respectively. During the vegetative stages, all seven Bt corn hybrids were
highly efficacious against Cry1Ab-susceptible and -heterozygous genotypes of
sugarcane borer, with a larval survival rate of <2% for the Bt-susceptible
genotype and < or =5% for the heterozygotes. However, 8-18% of the heterozygous
genotype survived on reproductive stage plants for four of the seven Bt corn
hybrids tested. The variation in performance of Bt corn cultivars at vegetative
and reproductive growth stages against Cry1Ab resistant sugarcane borer suggests
differential seasonal expression that may hasten resistance in the field. Bt corn
hybrids expressing a "high dose" for European corn borer, Ostrinia nubilalis
(Hö¼bner), may not produce a sufficient high dose for the sugarcane borer.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18232406 [PubMed - indexed for MEDLINE]

86: J Econ Entomol. 2007 Dec;100(6):1862-70.

DiPel-selected Ostrinia nubilalis larvae are not resistant to transgenic corn
expressing Bacillus thuringiensis Cry1Ab.

Li H, Buschman LL, Huang F, Zhu KY, Bonning B, Oppert B.

Department of Entomology, 123 Waters Hall, Kansas State University, Manhattan, KS
66506, USA.

The survival of KS-SC DiPel-resistant and -susceptible European corn borer,
Ostrinia nubilalis (Hö¼bner), was evaluated on different tissues from corn, Zea
mays L., hybrids, including a nontransgenic and two transgenic corn plants
(events MON810 and Bt11) expressing high doses of Bacillus thuringiensis (Bt)
Cry1Ab. The survival of Bt-resistant and -susceptible third instars was similar
after a 5-d exposure to transgenic plant tissues. Survivors eventually died when
returned to Bt corn tissues, but many were able to continue development when
transferred to non-Bt corn tissues. Survival of resistant and susceptible larvae
also was evaluated in bioassays with dilutions of leaf extracts from the three
corn hybrids incorporated in an artificial diet. In these assays, survival was
significantly higher for resistant O. nubilalis neonates at three of the five
dilutions compared with the susceptible strain, but the resistance ratio was only
2.2- and 2.4-fold for MON810 and Bt11, respectively. The data demonstrate that
Bt-resistant and unselected control O. nubilalis larvae were similar in
susceptibility to MON810 and Bt11 event corn hybrids. Although we were unable to
evaluate the Cry1Ab protein that larvae were exposed to in the transgenic tissue
because of company restrictions, Cry1Ab protoxin produced in Escherichia coli was
incubated with extracts from non-Bt corn leaves to simulate the in planta effect
on the transgenic protein. Cry1Ab protoxin was hydrolyzed rapidly by enzymes in
the corn extract into peptide fragments with molecular masses ranging from 132 to
74 kDa, and eventually 58 kDa. Overall, these data suggest that plant enzymes
hydrolyze transgenic toxin to one that is functionally activated. Therefore,
resistant insect populations with reduced proteinase activity do not seem to pose
a threat to the efficacy of commercial MON810 and Bt11 corn hybrids.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18232404 [PubMed - indexed for MEDLINE]

87: Bull Acad Natl Med. 2007 Apr-May;191(4-5):807-14; discussion 814.

[Allergic risk and role of the Allergy Vigilance Network]

[Article in French]

Moneret-Vautrin DA.

Mö©decine interne, immunologie clinique et allergologique, Hö´pital Central, 54035
Nancy cedex.

The recent increase in the incidence of severe anaphylaxis calls for continual
assessment of risk factor and dangers associated with food allergy, keeping
abreast of changes in the food industry. Allergologists, regulatory bodies and
the food industry are all responsible for food safety. The Allergy Vigilance
Network, founded by a university research team and comprising 398 French and
Belgian allergologists, has developed a three-point strategy. First, reporting
cases of severe anaphylaxis of document allergic origin makes it possible to
monitor the prevalence of food allergens and to evaluate the quality of
management of allergy-related emergencies, thus providing data suitable for
estimating the economic burden of anaphylaxis. The second objective of the
network is to set up multicenter trials to determine the prevalence of
sensitization to risk allergens, such as peanut, lupin and plant pollen, of which
transgenic varieties will soon emerge. The third objective is screening and
long-term monitoring of dangers related to new foods, ingredients and adjuvant
sensitizing factors. Post-marketing monitoring of potential allergic risks
arising from genetically modified food is another aim of the network, together
with the establishment of a serum bank, following WHO/FAO recommendations. The
Allergy Vigilance Network, together with the French National Institute for Food
Safety (AFSSA), the Ministry of Consumer Affairs (DGCCRF) and various patient
associations, is striving to analyse and deal with dangers related to the
allergenicity of natural and modified food proteins.

Publication Types:
Comparative Study
English Abstract

PMID: 18225435 [PubMed - indexed for MEDLINE]

88: Plant Mol Biol. 2008 Mar;66(5):551-63. Epub 2008 Jan 26.

Genome-wide allele-specific expression analysis using Massively Parallel
Signature Sequencing (MPSS) reveals cis- and trans-effects on gene expression in
maize hybrid meristem tissue.

Guo M, Yang S, Rupe M, Hu B, Bickel DR, Arthur L, Smith O.

Pioneer Hi-Bred International, Inc., A DuPont Business, Johnston, IA, 50131-0552,
USA. mei.guo@pioneer.com

Allelic differences in expression are important genetic factors contributing to
quantitative trait variation in various organisms. However, the extent of
genome-wide allele-specific expression by different modes of gene regulation has
not been well characterized in plants. In this study we developed a new
methodology for allele-specific expression analysis by applying Massively
Parallel Signature Sequencing (MPSS), an open ended and sequencing based mRNA
profiling technology. This methodology enabled a genome-wide evaluation of cis-
and trans-effects on allelic expression in six meristem stages of the maize
hybrid. Summarization of data from nearly 400 pairs of MPSS allelic signature
tags showed that 60% of the genes in the hybrid meristems exhibited differential
allelic expression. Because both alleles are subjected to the same trans-acting
factors in the hybrid, the data suggest the abundance of cis-regulatory
differences in the genome. Comparing the same allele expressed in the hybrid
versus its inbred parents showed that 40% of the genes were differentially
expressed, suggesting different trans-acting effects present in different
genotypes. Such trans-acting effects may result in gene expression in the hybrid
different from allelic additive expression. With this approach we quantified gene
expression in the hybrid relative to its inbred parents at the allele-specific
level. As compared to measuring total transcript levels, this study provides a
new level of understanding of different modes of gene regulation in the hybrid
and the molecular basis of heterosis.

PMID: 18224447 [PubMed - indexed for MEDLINE]

89: Trends Biotechnol. 2008 Mar;26(3):139-45. Epub 2008 Jan 28.

Metabolic engineering of carotenoid biosynthesis in plants.

Giuliano G, Tavazza R, Diretto G, Beyer P, Taylor MA.

Ente per le Nuove tecnologie, l'Energia e l'Ambiente (ENEA), Casaccia Research
Center, PO Box 2400, 00123 S.M. di Galeria (Roma), Italy.
giuliano@casaccia.enea.it

Carotenoids are one of the most diverse classes of natural compounds. Plant
carotenoids are composed of a C40 isoprenoid skeleton with or without epoxy,
hydroxy and keto groups. They have fundamental roles in human nutrition as
antioxidants and vitamin A precursors and their consumption is increasingly
associated with protection from a range of diseases. They are also used
commercially as safe food, feed and cosmetic colorants and they protect plants
from photooxidative stress. In the past six years many metabolic engineering
efforts have been undertaken in plants aiming to improve the nutritional value of
staple crops, to enable the use of plants as 'cell factories' for producing
specialty carotenoids and to improve plant resistance to abiotic stress.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 18222560 [PubMed - indexed for MEDLINE]

90: Ann Allergy Asthma Immunol. 2008 Jan;100(1 Suppl 2):S23-9.

Hereditary angiodema: a current state-of-the-art review, VI: novel therapies for
hereditary angioedema.

Frank MM.

Department of Pediatrics, Duke University School of Medicine, Durham, NC 27710,
USA. frank007@mc.duke.edu

OBJECTIVE: To provide a comprehensive overview on clinical trial design and
results of emerging therapies for the treatment of hereditary angioedema (HAE).
DATA SOURCES: MEDLINE or PubMed literature searches were conducted to identify
double-blind, placebo-controlled trials investigating C1 esterase replacement,
kallikrein inhibitor, and bradykinin receptor 2 antagonist therapies. STUDY
SELECTION: Ongoing trials or those just recently completed from all companies
developing a product for the treatment of HAE are discussed. RESULTS: All of
these agents are believed to be effective when tested in patients in phase 1 or
phase 2 trials. The studies have many features in common, including being
placebo-controlled and blinded; having a preliminary screening visit at which the
diagnosis is confirmed; having either low circulating C1 inhibitor protein levels
or low levels of functional C1 inhibitor, low C4 levels, and normal C1q levels;
enrolling individuals who are relatively early in attacks (4-6 hours from the
onset); and stipulating that patients continue taking the medications that they
have been taking in the long term. The type of attack acceptable for each
treatment protocol varies from study to study. Some allow peripheral edema
attacks, some facial attacks, and in some studies, the Food and Drug
Administration has allowed purified serum C1 inhibitor to be used as a rescue
medication if the patient remains in difficulty after the study drug has been
used and found to be ineffective. CONCLUSION: The outlook for new, effective
short-term therapy appears to be excellent. In the near future, a whole new
therapeutic armamentarium to care for patients with HAE should be available in
the United States.

Publication Types:
Review

PMID: 18220149 [PubMed - indexed for MEDLINE]

91: Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):E10; author reply E11. Epub 2008
Jan 24.

Comment on:
Proc Natl Acad Sci U S A. 2007 Oct 9;104(41):16204-8.

Study of Bt impact on caddisflies overstates its conclusions: response to
Rosi-Marshall et al.

Parrott W.

Publication Types:
Comment
Letter

PMID: 18218776 [PubMed - indexed for MEDLINE]

92: J Dairy Sci. 2008 Feb;91(2):673-8.

Alfalfa containing the glyphosate-tolerant trait has no effect on feed intake,
milk composition, or milk production of dairy cattle.

Combs DK, Hartnell GF.

Department of Dairy Science, University of Wisconsin, Madison, Wisconsin 53706,
USA. dkcombs@wisc.edu

The objective of this experiment was to assess if feeding glyphosate-tolerant
alfalfa affects feed intake, milk composition, or milk production of dairy cows.
One alfalfa (Medicago sativa), variety expressing the CP4 EPSPS protein and grown
in southeastern Washington State was harvested at the late vegetative stage as
hay. Three commercial conventional varieties of alfalfa hay of similar nutrient
composition and harvested in the same geographic region were fed to cows as
controls. The commercial hays were selected to be similar in crude protein [18%
of dry matter (DM)] and neutral detergent fiber (40% of DM) to the
glyphosate-tolerant hay. Sixteen multiparous Holstein cows were fed diets
containing alfalfa hay (39.7% of diet DM) from either the glyphosate-tolerant
alfalfa, or 1 of the 3 conventional varieties. Diets contained at least 15.7%
crude protein and 29% neutral detergent fiber. Experimental design was a
replicated 4 x 4 Latin square. Periods were 28 d and feed intake, milk yield, and
milk composition were summarized over the last 14 d of each period. Daily milk
yield (38.0 kg) and 4% fat-corrected milk (34.7 kg) were not affected by
treatment. Milk fat (3.44%) and milk true protein (2.98%) were also not affected
by source of hay. Milk lactose (4.72%) and soldis-not-fat (8.5%) did not differ
due to treatment. Dry matter intake was similar across treatments (24.4 kg/d).
These results are consistent with data from feeding trials with other
glyphosate-tolerant crops and previously reported compositional comparisons of
glyphosate-tolerant alfalfa with controls. Milk production, milk composition,
feed intake, and feed efficiency were not affected by feeding diets that
contained nearly 40% glyphosate-tolerant alfalfa hay to lactating dairy cows.

PMID: 18218755 [PubMed - indexed for MEDLINE]

93: J Agric Food Chem. 2008 Feb 27;56(4):1259-63. Epub 2008 Jan 25.

Herbicide-induced anthocyanin accumulation in transgenic rice by expression of
rice OSB2 under the control of rice CYP72A21 promoter.

Hirose S, Kawahigashi H, Tagiri A, Ohkawa Y.

National Institute of Agrobiological Sciences, Ibaraki, Japan.

CYP72A21, a rice cytochrome P450 gene, is induced by chloroacetamide herbicides.
OSB2, a rice myc-type transcription factor, induces anthocyanin accumulation in
rice leaves. To produce plants for biomonitoring by color change, we combined the
CYP72A21 promoter and the OSB2 gene and introduced them into the rice isogenic
line Taichung-65 CB A (T65), which contains loci CB and A from the rice cultivar
Murasakiine. Leaves of the transgenic plants turned red upon treatment with the
chloroacetamide herbicides acetochlor, alachlor, and metolachlor. Seedling shoots
reddened upon treatment with alachlor or metolachlor at 10 microM, a
concentration slightly higher than that used in the field. Anthocyanin content
was increased approximately 200% by the treatment. The color changes were
consistent with increased shoot expression of OSB2 and the anthocyanidin synthase
gene (ANS). This system promises easy detection of rice plant gene expression.
Transgenic plants could be used in the future to biomonitor accumulated
herbicides.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18217708 [PubMed - indexed for MEDLINE]

94: Adv Med Sci. 2007;52:98-103.

Food allergies, cross-reactions and agroalimentary biotechnologies.

Ronchetti R, Kaczmarski MG, Haå‚uszka J, Jesenak M, Villa MP.

Department of Paediatrics, 2nd School of Medicine, University La Sapienza, S.
Andrea Hospital, Rome, Italy. roberto.ronchetti@ospedalesantandrea.it

The discrepancy between what the general public and specialist in allergic
diseases regard as a true food allergy can in part depend on the frequent
evidence of subjects in whom clinical symptoms elicited by a given food allergen
are frequently not reproducible: this suggests the existence of allergens
variably present in certain foods. In adults and older children common is a form
of food allergy associated with inhaled allergens, especially pollens. In this
allergic form pollens and various vegetal food often cross react but the
underlying scientific rationale is largely unclear. From the study of the
"latex-fruits allergic syndrome" and the "oral allergic syndrome" emerged that
the cross reactivity depends on epitopes of pollens and vegetables belonging to
one of the 14 classes of the "pathogenesis related proteins" (PRPs). Vegetables
produce PRPs in response to infection or after plant injury or application of
chemicals: long-term conservation and methods used for rapid artificial ripening
of vegetables can cause plant to produce PRPs or other allergens. A genetic
selection of vegetables "protecting themselves against infection and infestation"
by mean of PRPs production is practiced in agroalimentary biotechnology. We deem
it urgent that the two realms, Medical Science (Allergology) and Agricultural
Biotechnology begin to communicate openly in order to produce food as efficiently
as possible but without harming the large part of the population which is
predisposed to allergy and react to PRPs.

Publication Types:
Review

PMID: 18217398 [PubMed - indexed for MEDLINE]

95: Proc Natl Acad Sci U S A. 2008 Jan 29;105(4):1339-42. Epub 2008 Jan 23.

A Caenorhabditis elegans allatostatin/galanin-like receptor NPR-9 inhibits local
search behavior in response to feeding cues.

Bendena WG, Boudreau JR, Papanicolaou T, Maltby M, Tobe SS, Chin-Sang ID.

Department of Biology, Queen's University, Kingston, ON, Canada.
bendenaw@biology.queensu.ca

Movement in Caenorhabditis elegans is the result of sensory cues creating
stimulatory and inhibitory output from sensory neurons. Four interneurons (AIA,
AIB, AIY, and AIZ) are the primary recipients of this information that is further
processed en route to motor neurons and muscle contraction. C. elegans has >1,000
G protein-coupled receptors (GPCRs), and their contribution to sensory-based
movement is largely undefined. We show that an allatostatin/galanin-like GPCR
(NPR-9) is found exclusively in the paired AIB interneuron. AIB interneurons are
associated with local search/pivoting behavior. npr-9 mutants display an
increased local search/pivoting that impairs their ability to roam and travel
long distances on food. With impaired roaming behavior on food npr-9 mutants
accumulate more intestinal fat as compared with wild type. Overexpression of
NPR-9 resulted in a gain-of-function phenotype that exhibits enhanced forward
movement with lost pivoting behavior off food. As such the animal travels a great
distance off food, creating arcs to return to food. These findings indicate that
NPR-9 has inhibitory effects on the AIB interneuron to regulate foraging
behavior, which, in turn, may affect metabolic rate and lipid storage.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18216257 [PubMed - indexed for MEDLINE]

96: Toxicology. 2008 Mar 12;245(1-2):24-34. Epub 2007 Dec 17.

Immunotoxicological studies of genetically modified rice expressing PHA-E lectin
or Bt toxin in Wistar rats.

Kroghsbo S, Madsen C, Poulsen M, Schrö¸der M, Kvist PH, Taylor M, Gatehouse A, Shu
Q, Knudsen I.

Department of Toxicology and Risk Assessment, National Food Institute, Technical
University of Denmark, Mö¸rkhö¸j Bygade 19, DK-2860 Sö¸borg, Denmark.
sck@food.dtu.dk

As part of the SAFOTEST project the immunmodulating effect of Cry1Ab protein from
Bacillus thuringiensis (Bt) and PHA-E lectin from kidney bean (Phaseolus vulgaris
erythroagglutinin) was examined in 28- and 90-day feeding studies in Wistar rats.
PHA-E lectin was chosen as positive control. Rats were fed control rice,
transgenic rice expressing Cry1Ab protein or PHA-E lectin, or transgenic rice
spiked with the purified recombinant protein. Total immunoglobulin levels,
mitogen-induced cell proliferation, T-dependent antibody response to sheep red
blood cells and the antigen-specific antibody response in serum were examined at
the end of the studies. A dose-dependent increase in mesenteric lymph node weight
and total immunoglobulin A was seen when feeding PHA-E transgenic rice alone or
spiked with 0.1% purified PHA-E lectin for 90 days indicating a local effect of
PHA-E in the intestine. No adverse effects of Cry1Ab protein were found. An
anti-PHA-E and anti-Cry1Ab antibody response was induced both after inhalation
(control groups) and after inhalation/ingestion (groups fed recombinant protein
alone or together with transgenic rice). In conclusion, only PHA-E lectin was
found to have an immunomodulating effect when feeding rats for 90 days with
approximately 70 mg PHA-E/kg bodyweight per day. As both PHA-E lectin and Cry1Ab
protein were capable of inducing an antigen-specific antibody response it is
important to make careful considerations when designing future animal studies to
avoid intake of proteins from the other groups by inhalation as well as to
examine the sensitization and elicitation potential of 'foreign' proteins before
introduction to the world market.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18215453 [PubMed - indexed for MEDLINE]

97: J Food Sci. 2008 Jan;73(1):S64-9.

Comparative physicochemical properties and structure of rice containing the
sck+cryIAc genes and its nontransgenic counterpart.

Li X, Huang K, Zhu B, Liang Z, Wei L, Luo Y.

College of Food Science and Nutritional Engineering, China Agricultural
University, Beijing 100083, China.

The physicochemical properties and structure of an insect-resistant rice,
Liangyou Kefeng Nr. 6 (IRR), containing the sck and cryIAc genes were compared
with those of its nontransgenic counterpart designated as Liangyou 2186
(control), considering their key role in determining commercial value. Basically
the appearance of IRR was not affected in terms of size and shape after foreign
gene transformation but improved with lower chalkiness. The milling yield of IRR
with lower chalkiness was higher measured by head rice yield compared with its
parental control. The differences in appearance and milling quality were
confirmed by the structure of raw rice grain by scanning electron microscopy
(SEM). Slight differences were observed in pasting properties and textural
quality determined by rapid viscosity analyzer and texture analyzer which was in
agreement with the result of the structure of cooked rice grain by SEM. The above
differences might be as a result of a positional effect of T-DNA insertion. On
the whole, the appearance, milling quality, and eating quality of IRR were not
adversely affected by transgenes, which will facilitate its acceptance by the
consumer after commercialization.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18211372 [PubMed - indexed for MEDLINE]

98: New Phytol. 2008;178(1):80-91. Epub 2008 Jan 16.

Ectopic expression of GmPAP3 alleviates oxidative damage caused by salinity and
osmotic stresses.

Li WY, Shao G, Lam HM.

Department of Biology, The Chinese University of Hong Kong, Shatin, NT, Hong Kong
SAR, China.

The primary biochemical reaction of purple acid phosphatases (PAP) is to catalyze
the hydrolysis of phosphate esters and anhydrides. However, the soybean GmPAP3
gene expression is induced by NaCl, osmotic, and oxidative treatments, indicating
a possible role of PAP in abiotic stress responses. Confocal and electron
microscopic studies demonstrated that GmPAP3 protein is mainly localized in
mitochondria, a primary site for reactive oxygen species (ROS) production. When
subjected to NaCl and polyethylene glycol (PEG) treatments, ectopic expression of
GmPAP3 in transgenic tobacco BY-2 cells mimicked the protective effects exhibited
by the antioxidant ascorbic acid: increase in the percentage of cells with active
mitochondria; reduction in the percentage of dead cells; and reduced accumulation
of ROS. In addition, when GmPAP3 transgenic Arabidopsis thaliana seedlings were
subjected to NaCl, PEG, and paraquat (PQ) treatments, the percentage of root
elongation was significantly higher than the wild type. Furthermore, PQ-induced
lipid peroxidation in these transgenic seedlings was also reduced. In summary,
the mitochondrial localized GmPAP3 may play a role in stress tolerance by
enhancing ROS scavenging.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18208471 [PubMed - indexed for MEDLINE]

99: Ecotoxicol Environ Saf. 2008 Jun;70(2):327-33. Epub 2008 Feb 21.

Does Cry1Ab protein affect learning performances of the honey bee Apis mellifera
L. (Hymenoptera, Apidae)?

Ramirez-Romero R, Desneux N, Decourtye A, Chaffiol A, Pham-Delö¨gue MH.

Instituto de Ecologia A.C., Km. 2.5 Carretera Antigua a Coatepec No. 351 El Haya,
91070 Xalapa, Veracruz, Mexico.

Genetically modified Bt crops are increasingly used worldwide but side effects
and especially sublethal effects on beneficial insects remain poorly studied.
Honey bees are beneficial insects for natural and cultivated ecosystems through
pollination. The goal of the present study was to assess potential effects of two
concentrations of Cry1Ab protein (3 and 5000ppb) on young adult honey bees.
Following a complementary bioassay, our experiments evaluated effects of the
Cry1Ab on three major life traits of young adult honey bees: (a) survival of
honey bees during sub-chronic exposure to Cry1Ab, (b) feeding behaviour, and (c)
learning performance at the time that honey bees become foragers. The latter
effect was tested using the proboscis extension reflex (PER) procedure. The same
effects were also tested using a chemical pesticide, imidacloprid, as positive
reference. The tested concentrations of Cry1Ab protein did not cause lethal
effects on honey bees. However, honey bee feeding behaviour was affected when
exposed to the highest concentration of Cry1Ab protein, with honey bees taking
longer to imbibe the contaminated syrup. Moreover, honey bees exposed to 5000ppb
of Cry1Ab had disturbed learning performances. Honey bees continued to respond to
a conditioned odour even in the absence of a food reward. Our results show that
transgenic crops expressing Cry1Ab protein at 5000ppb may affect food consumption
or learning processes and thereby may impact honey bee foraging efficiency. The
implications of these results are discussed in terms of risks of transgenic Bt
crops for honey bees.

PMID: 18206234 [PubMed - in process]

100: Plant Mol Biol. 2008 Mar;66(5):445-62. Epub 2008 Jan 18.

Overexpression of OsiSAP8, a member of stress associated protein (SAP) gene
family of rice confers tolerance to salt, drought and cold stress in transgenic
tobacco and rice.

Kanneganti V, Gupta AK.

Department of Plant Biotechnology, Madurai Kamaraj University, Madurai, 625021,
TamilNadu, India.

We describe here the isolation and characterization of OsiSAP8, a member of
stress Associated protein (SAP) gene family from rice characterized by the
presence of A20 and AN1 type Zinc finger domains. OsiSAP8 is a multiple stress
inducible gene, induced by various stresses, namely heat, cold, salt,
desiccation, submergence, wounding, heavy metals as well as stress hormone
Abscisic acid. OsiSAP8 protein fused to GFP was localized towards the periphery
of the cells in the epidermal cells of infiltrated Nicotiana benthamiana leaves.
Yeast two hybrid analysis revealed that A20 and AN1 type zinc-finger domains of
OsiSAP8 interact with each other. Overexpression of the gene in both transgenic
tobacco and rice conferred tolerance to salt, drought and cold stress at seed
germination/seedling stage as reflected by percentage of germination and gain in
fresh weight after stress recovery. Transgenic rice plants were tolerant to salt
and drought during anthesis stage without any yield penalty as compared to
unstressed transgenic plants.

PMID: 18205020 [PubMed - indexed for MEDLINE]

101: Shokuhin Eiseigaku Zasshi. 2007 Dec;48(6):170-8.

[Development and evaluation of qualitative detection methods for unapproved
genetically modified rice (LLRice)]

[Article in Japanese]

Watanabe T, Shiramasa Y, Furui S, Kitta K, Minegishi Y, Akiyama H, Maitani T.

National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo
158-8501, Japan.

We developed a specific method to extract DNA from rice grain samples and
modified the qualitative real-time PCR method provided by Bayer Co., Ltd. for
reliable detection of the genetically modified (GM) rice variety, LLRice601,
which has not undergone safety assessment for regulatory approval in Japan.
Moreover, we conducted a data analysis to confirm the results obtained with
real-time PCR. The yields of DNA extracted from powdered samples of rice grains
were almost equal among 5 different varieties of rice, and there was no
significant difference in the yield over three days. Reliable results were
obtained using 50 ng of the extracted DNA as the template for real-time PCR. To
examine the adequacy of the methods, we organized an interlaboratory study with
the participation of 2 laboratories, in which 80 test samples were analyzed in a
blinded manner. The statistical analysis revealed no significant difference in
the Ct value for the endogenous gene of the DNA samples and for the targeted DNA
sequence of 0.1% samples. The limit of detection of the method was approximately
0.1%. Analysis of the fluorescence intensity of the PCR-amplified product of the
construct-specific DNA sequence suggested that it may be reasonable to judge a
sample as positive when a Ct value of less than 40 is obtained.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 18203502 [PubMed - indexed for MEDLINE]

102: Proc Natl Acad Sci U S A. 2008 Feb 5;105(5):1431-5. Epub 2008 Jan 17.

Comment in:
Proc Natl Acad Sci U S A. 2008 Feb 12;105(6):1777-8.

Nutritional impact of elevated calcium transport activity in carrots.

Morris J, Hawthorne KM, Hotze T, Abrams SA, Hirschi KD.

Vegetable and Fruit Improvement Center, Texas A&M University, College Station, TX
77845, USA.

Nutrition recommendations worldwide emphasize ingestion of plant-based diets
rather than diets that rely primarily on animal products. However, this
plant-based diet could limit the intake of essential nutrients such as calcium.
Osteoporosis is one of the world's most prevalent nutritional disorders, and
inadequate dietary calcium is a known contributor to the pathophysiology of this
condition. Previously, we have modified carrots to express increased levels of a
plant calcium transporter (sCAX1), and these plants contain approximately
2-fold-higher calcium content in the edible portions of the carrots. However, it
was unproven whether this change would increase the total amount of bioavailable
calcium. In randomized trials, we labeled these modified carrots with isotopic
calcium and fed them to mice and humans to assess calcium bioavailability. In
mice feeding regimes (n = 120), we measured (45)Ca incorporation into bones and
determined that mice required twice the serving size of control carrots to obtain
the calcium found in sCAX1 carrots. We used a dual-stable isotope method with
(42)Ca-labeled carrots and i.v. (46)Ca to determine the absorption of calcium
from these carrots in humans. In a cross-over study of 15 male and 15 female
adults, we found that when people were fed sCAX1 and control carrots, total
calcium absorption per 100 g of carrots was 41% +/- 2% higher in sCAX1 carrots.
Both the mice and human feeding studies demonstrate increased calcium absorption
from sCAX1-expressing carrots compared with controls. These results demonstrate
an alternative means of fortifying vegetables with bioavailable calcium.

Publication Types:
Randomized Controlled Trial
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18202180 [PubMed - indexed for MEDLINE]

103: Proc Natl Acad Sci U S A. 2008 Jan 22;105(3):859-64. Epub 2008 Jan 16.

The intersection between cell wall disassembly, ripening, and fruit
susceptibility to Botrytis cinerea.

Cantu D, Vicente AR, Greve LC, Dewey FM, Bennett AB, Labavitch JM, Powell AL.

Departments of Plant Sciences and Viticulture and Enology, University of
California, Davis, CA 95616, USA.

Fruit ripening is characterized by processes that modify texture and flavor but
also by a dramatic increase in susceptibility to necrotrophic pathogens, such as
Botrytis cinerea. Disassembly of the major structural polysaccharides of the cell
wall (CW) is a significant process associated with ripening and contributes to
fruit softening. In tomato, polygalacturonase (PG) and expansin (Exp) are among
the CW proteins that cooperatively participate in ripening-associated CW
disassembly. To determine whether endogenous CW disassembly influences the
ripening-regulated increase in necrotropic pathogen susceptibility, B. cinerea
susceptibility was assessed in transgenic fruit with suppressed polygalacturonase
(LePG) and expansin (LeExp1) expression. Suppression of either LePG or LeExp1
alone did not reduce susceptibility but simultaneous suppression of both
dramatically reduced the susceptibility of ripening fruit to B. cinerea, as
measured by fungal biomass accumulation and by macerating lesion development.
These results demonstrate that altering endogenous plant CW disassembly during
ripening influences the course of infection by B. cinerea, perhaps by changing
the structure or the accessibility of CW substrates to pathogen CW-degrading
enzymes. Recognition of the role of ripening-associated CW metabolism in
postharvest pathogen susceptibility may be useful in the design and development
of strategies to limit pathogen losses during fruit storage, handling, and
distribution.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18199833 [PubMed - indexed for MEDLINE]

104: Mol Genet Genomics. 2008 Mar;279(3):291-301. Epub 2008 Jan 16.

Cloning and characterization of three genes encoding Qb-SNARE proteins in rice.

Bao YM, Wang JF, Huang J, Zhang HS.

State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing
Agricultural University, Nanjing, 210095, China.

Qb-SNARE proteins belong to the superfamily of SNAREs (soluble
N-ethylmaleimide-sensitive factor attachment protein receptors) and function as
important components of the vesicle trafficking machinery in eukaryotic cells.
Here, we report three novel plant SNARE (NPSN) genes isolated from rice and named
OsNPSN11, OsNPSN12 and OsNPSN13. They have about 70% nucleotide identity over
their entire coding regions and similar genomic organization with ten exons and
nine introns in each gene. Multiple alignment of deduced amino acid sequences
indicate that the OsNPSNs proteins are homologous to AtNPSNs from Arabidopsis,
containing a Qb-SNARE domain and a membrane-spanning domain in the C-terminal
region. Semi-quantitative RT-PCR assays showed that the OsNPSNs were ubiquitously
and differentially expressed in roots, culms, leaves, immature spikes and
flowering spikes. The expression of OsNPSNs was significantly activated in rice
seedlings treated with H(2)O(2), but down-regulated under NaCl and PEG6000
stresses. Transient expression method in onion epidermal cells revealed that
OsNPSNs were located in the plasma membrane. Transformed yeast cells with OsNPSNs
had better growth rates than empty-vector transformants when cultured on either
solid or liquid selective media containing various concentrations of H(2)O(2),
but more sensitive to NaCl and mannitol stresses. The 35S:OsNPSN11 transgenic
tobacco also showed more tolerance to H(2)O(2) and sensitivity to NaCl and
mannitol than non-transgenic tobacco. These results indicate that OsNPSNs may be
involved in different aspects of the signal transduction in plant and yeast
responses to abiotic stresses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18197419 [PubMed - indexed for MEDLINE]

105: Traffic. 2008 Mar;9(3):408-16. Epub 2007 Jan 9.

Lack of a vacuolar sorting receptor leads to non-specific missorting of soluble
vacuolar proteins in Arabidopsis seeds.

Craddock CP, Hunter PR, Szakacs E, Hinz G, Robinson DG, Frigerio L.

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.

The plant vacuolar sorting receptor (VSR) binds proteins carrying vacuolar
sorting signals (VSS) of the 'sequence-specific' type (ssVSS) but not the
C-terminal, hydrophobic sorting signals (ctVSS). Seeds of Arabidopsis mutants
lacking the major VSR isoform, AtVSR1, secrete a proportion of the proteins
destined to storage vacuoles. The sorting signals for these proteins are not well
defined, but they do not seem to be of the ssVSS type. Here, we tested whether
absence of VSR1 in seeds leads to secretion of reporter proteins carrying ssVSS
but not ctVSS. Our results show that reporters carrying either ssVSS or ctVSS are
equally secreted in the absence of VSR1. We discuss our findings in relation to
the current model for vacuolar sorting.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18194392 [PubMed - indexed for MEDLINE]

106: New Phytol. 2008;178(1):147-56. Epub 2008 Jan 10.

Effects of host plant environment and Ustilago maydis infection on the fungal
endophyte community of maize (Zea mays).

Pan JJ, Baumgarten AM, May G.

Department of Biology, The University of Akron, Akron, OH 44325, USA.
jepan@uakron.edu

The focus of many fungal endophyte studies has been how plants benefit from
endophyte infection. Few studies have investigated the role of the host plant as
an environment in shaping endophyte community diversity and composition. The
effects that different attributes of the host plant, that is, host genetic
variation, host variation in resistance to the fungal pathogen Ustilago maydis
and U. maydis infection, have on the fungal endophyte communities in maize (Zea
mays) was examined. The internal transcribed spacer (ITS) region of the rDNA was
sequenced to identify fungi and the endophyte communities were compared in six
maize lines that varied in their resistance to U. maydis. It was found that host
genetic variation, as determined by maize line, had significant effects on
species richness, while the interactions between line and U. maydis infection and
line and field plot had significant effects on endophyte community composition.
However, the effects of maize line were not dependent on whether lines were
resistant or susceptible to U. maydis. Almost 3000 clones obtained from 58 plants
were sequenced to characterize the maize endophyte community. These results
suggest that the endophyte community is shaped by complex interactions and
factors, such as inoculum pool and microclimate, may be important.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18194146 [PubMed - indexed for MEDLINE]

107: J AOAC Int. 2007 Nov-Dec;90(6):1517-25.

Detection and characterization of cry1Ac transgene construct in Bt cotton:
multiple polymerase chain reaction approach.

Singh CK, Ojha A, Kachru DN.

Industrial Toxicology Research Centre, Post Box No. 80, M.G. Marg Lucknow-226001
U.P., India.

To comply with international labeling regulations for genetically modified (GM)
crops and food, and to enable proper identification of GM organisms (GMOs),
effective methodologies and reliable approaches are needed. The spurious and
unapproved GM planting has contributed to crop failures and commercial losses. To
ensure effective and genuine GM cultivation, a methodology is needed to detect
and identify the trait of interest and concurrently evaluate the structural and
functional stability of the transgene insert. A multiple polymerase chain
reaction (PCR) approach was developed for detection, identification, and gene
stability confirmation of cry1Ac transgene construct in Bt cotton. As many as 9
samples of Bt cotton hybrid seeds comprising 3 approved Bt hybrids, MECH-12Bt,
MECH-162Bt, MECH-184Bt, and a batch of 6 nonapproved Bt hybrids were tested.
Initially, single standard PCR assays were run to amplify predominant GM DNA
sequences (CaMV 35S promoter, nos terminator, and npt-II marker gene); a
housekeeping gene, Gossypium hirsutum fiber-specific acyl carrier protein gene
(acp1); a trait-specific transgene (cry1Ac); and a sequence of 7S 3'
transcription terminator which specifically borders with 3' region of cry1Ac
transgene cassette. The concurrent amplification of all sequences of the entire
cassette was performed by 3 assays, duplex, triplex, and quadruplex multiplex PCR
assays, under common assay conditions. The identity of amplicons was reconfirmed
by restriction endonuclease digestion profile. The 2 distinct transgene
cassettes, cry1Ac and npt-II, of the Bt cotton were amplified using the
respective forward primer of promoter and reverse primer of terminator. The
resultant amplicons were excised, eluted, and purified. The purified amplicons
served as template for nested PCR assays. The nested PCR runs confirmed the
transgene construct orientation and identity. The limit of detection as
established by our assay for GM trait (cry1Ac) was 0.1%. This approach can be
adopted as a standard procedure for complete molecular characterization of Bt
cotton. These assays will be of interest and use to importers, breeders, research
laboratories, safety regulators, and food processors for detection of cry1Ac
bearing GMOs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18193727 [PubMed - indexed for MEDLINE]

108: Trends Biotechnol. 2008 Feb;26(2):77-81. Epub 2008 Jan 11.

Potential for metabolic engineering of resveratrol biosynthesis.

Halls C, Yu O.

Donald Danforth Plant Science Center, 975 North Warson Road, Saint Louis, MO
63132, USA.

Resveratrol, an interesting plant phenolic compound, is found in red wine but is
not widely distributed in other common food sources. Health benefits of
resveratrol include prevention of cardiovascular diseases and cancers, and--as
discovered more recently--promotion of longevity in several animal systems. The
pathway and enzymes for resveratrol biosynthesis are well characterized.
Furthermore, metabolic engineering of this compound has been achieved in plants,
microbes and animals. This review attempts to summarize current understanding of
resveratrol pathway-engineering in various systems, to outline the challenges in
commercial applications and to identify future opportunities for resveratrol
bioengineering.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 18191264 [PubMed - indexed for MEDLINE]

109: Trends Biotechnol. 2008 Feb;26(2):64-9. Epub 2008 Jan 11.

Can GM sorghum impact Africa?

Botha GM, Viljoen CD.

GMO Testing Facility, University of the Free State, PO Box 339, Bloemfontein
9300, South Africa. bothagm.sci@mail.ufs.ac.za

It is said that genetic modification (GM) of grain sorghum has the potential to
alleviate hunger in Africa. To this end, millions of dollars have been committed
to developing GM sorghum. Current developments in the genetic engineering of
sorghum are similar to efforts to improve cassava and other traditional African
crops, as well as rice in Asia. On closer analysis, GM sorghum is faced with the
same limitations as 'Golden Rice' (GM rice) in the context of combating vitamin A
deficiency (VAD) efficiently and sustainably. Thus, it is questionable whether
the cost of developing GM sorghum can be justified when compared to the cost of
investing in sustainable agricultural practice in Africa.

Publication Types:
Review

PMID: 18191263 [PubMed - indexed for MEDLINE]

110: Tanpakushitsu Kakusan Koso. 2008 Jan;53(1):65-71.

[Mutational reconstructed ferric chelate reductase confers enhanced tolerance in
rice to iron deficiency in calcareous soil]

[Article in Japanese]

Ishimaru Y, Nishizawa NK.

aishimar@mail.ecc.u-tokyo.ac.jp

Publication Types:
Review

PMID: 18186305 [PubMed - indexed for MEDLINE]

111: Proc Natl Acad Sci U S A. 2008 Jan 15;105(2):470-5. Epub 2008 Jan 9.

A crop population perspective on maize seed systems in Mexico.

Dyer GA, Taylor JE.

Department of Agricultural and Resource Economics, University of California, One
Shields Avenue, Davis, CA 95616, USA. gdyer@primal.ucdavis.edu

Improvement of local germplasm through artificial selection is regarded as the
main force behind maize evolution and diversity in Mexico, the crop's center of
origin. This perspective neglects the larger social context of maize evolution.
Using a theoretical approach and Mexico-wide data, we show that farmer-led
evolution of maize is largely driven by a technological diffusion and
appropriation process that selectively integrates nonlocal germplasm into local
seed stocks. Our approach construes farmer practices as events in the life
history of seed to build a demographic model. The model shows how random and
systematic differences in management combine to structure maize seed populations
into subpopulations that can spread or become extinct, in some cases
independently of visible agronomic advantages. The process involves continuous
population bottlenecks that can lead to diversity loss. Nonlocal germplasm thus
might play a critical role in maintaining diversity in individual localities.
Empirical estimates show that introduction of nonlocal seed in Central and
Southeastern Mexico is rarer than previously thought; prompt replacement further
prevents new seed from spreading. Yet introduced seed perceived as valuable
diffuses rapidly, contributing variation in the form of type diversity or through
introgression into local seed. Maize seed dynamics and evolution are thus part of
a complex social process driven by farmers' desire to appropriate the value in
maize farming, not always achieved by preserving or improving local seed stocks.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18184814 [PubMed - indexed for MEDLINE]

112: Mol Plant Microbe Interact. 2008 Feb;21(2):171-7.

The expression of a bean PGIP in transgenic wheat confers increased resistance to
the fungal pathogen Bipolaris sorokiniana.

Janni M, Sella L, Favaron F, Blechl AE, De Lorenzo G, D'Ovidio R.

Dipartimento di Agrobiologia e Agrochimica, University of Tuscia, 01100 Viterbo,
Italy.

A possible strategy to control plant pathogens is the improvement of natural
plant defense mechanisms against the tools that pathogens commonly use to
penetrate and colonize the host tissue. One of these mechanisms is represented by
the host plant's ability to inhibit the pathogen's capacity to degrade plant cell
wall polysaccharides. Polygalacturonase-inhibiting proteins (PGIP) are plant
defense cell wall glycoproteins that inhibit the activity of fungal
endopolygalacturonases (endo-PGs). To assess the effectiveness of these proteins
in protecting wheat from fungal pathogens, we produced a number of transgenic
wheat lines expressing a bean PGIP (PvPGIP2) having a wide spectrum of
specificities against fungal PGs. Three independent transgenic lines were
characterized in detail, including determination of the levels of PvPGIP2
accumulation and its subcellular localization and inhibitory activity. Results
show that the transgene-encoded protein is correctly secreted into the apoplast,
maintains its characteristic recognition specificities, and endows the transgenic
wheat with new PG recognition capabilities. As a consequence, transgenic wheat
tissue showed increased resistance to digestion by the PG of Fusarium
moniliforme. These new properties also were confirmed at the plant level during
interactions with the fungal pathogen Bipolaris sorokiniana. All three lines
showed significant reductions in symptom progression (46 to 50%) through the
leaves following infection with this pathogen. Our results illustrate the
feasibility of improving wheat's defenses against pathogens by expression of
proteins with new capabilities to counteract those produced by the pathogens.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18184061 [PubMed - indexed for MEDLINE]

113: Adv Exp Med Biol. 2008;606:357-93.

Producing recombinant human milk proteins in the milk of livestock species.

Bö¶sze Z, Baranyi M, Whitelaw CB.

Agricultural Biotechnology Center, Gö¶dö¶llöµ, Hungary. bosze@abc.hu

Recombinant human proteins produced by the mammary glands of genetically modified
transgenic livestock mammals represent a special aspect of milk bioactive
components. For therapeutic applications, the often complex posttranslational
modifications of human proteins should be recapitulated in the recombinant
products. Compared to alternative production methods, mammary gland production is
a viable option, underlined by a number of transgenic livestock animal models
producing abundant biologically active foreign proteins in their milk.
Recombinant proteins isolated from milk have reached different phases of clinical
trials, with the first marketing approval for human therapeutic applications from
the EMEA achieved in 2006.

Publication Types:
Review

PMID: 18183938 [PubMed - indexed for MEDLINE]

114: Adv Exp Med Biol. 2008;606:345-56.

Manipulation of milk fat composition through transgenesis.

Van Eenennaam AL, Medrano JF.

Department of Animal Science, University of California, Davis 95616-8521, USA.
alvaneenennaam@ucdavis.edu

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Review

PMID: 18183937 [PubMed - indexed for MEDLINE]

115: Nat Biotechnol. 2008 Jan;26(1):73-81.

Erratum in:
Nat Biotechnol. 2008 Feb;26(2):241.

Allergenicity assessment of genetically modified crops--what makes sense?

Goodman RE, Vieths S, Sampson HA, Hill D, Ebisawa M, Taylor SL, van Ree R.

Department of Food Science & Technology, University of Nebraska, Lincoln,
Nebraska, 68583-0955, USA. rgoodman2@unlnotes.unl.edu

GM crops have great potential to improve food quality, increase harvest yields
and decrease dependency on certain chemical pesticides. Before entering the
market their safety needs to be scrutinized. This includes a detailed analysis of
allergenic risks, as the safety of allergic consumers has high priority. However,
not all tests currently being applied to assessing allergenicity have a sound
scientific basis. Recent events with transgenic crops reveal the fallacy of
applying such tests to GM crops.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 18183024 [PubMed - indexed for MEDLINE]

116: J Exp Bot. 2007;58(15-16):4147-59.

Expression of a RING-HC protein from rice improves resistance to Pseudomonas
syringae pv. tomato DC3000 in transgenic Arabidopsis thaliana.

Cheung MY, Zeng NY, Tong SW, Li FW, Zhao KJ, Zhang Q, Sun SS, Lam HM.

Department of Biology, The Chinese University of Hong Kong, Shatin, Hong Kong
SAR.

A cDNA clone (OsRHC1) was obtained, which encodes a novel RING zinc finger
protein sharing similar structural features (multiple transmembrane domains at
the N-half; a unique RING zinc finger consensus
Cys-X(2)-Cys-X(11)-Cys-X-His-X(3)-Cys-X(2)-Cys-X(6)-Cys-X(2)-Cys at the C
terminus) to a group of closely related annotated proteins from both monocots and
dicots. OsRHC1 was found to be localized on plasma membrane of rice cells and
induced by wounding in rice lines containing Xa loci. Ecotopic expression of the
OsRHC1 cDNA from rice (a monocot) in transgenic Arabidopsis thaliana (a dicot)
enhanced the defence response toward Pseudomonas syringae pv. tomato DC3000,
suggesting that OsRHC1 may confer broad-spectrum disease resistance. The
protective effects were neutralized in the presence of MG132 or in an npr1-3
mutation background, indicating that the function of OsRHC1 is dependent on the
ubiquitin-mediated protein degradation via the 26S proteasome and the presence of
the key defence response regulator NPR1.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18182423 [PubMed - indexed for MEDLINE]

117: Plant J. 2008 Apr;54(1):141-51. Epub 2008 Jan 7.

Overexpression of membrane-associated acyl-CoA-binding protein ACBP1 enhances
lead tolerance in Arabidopsis.

Xiao S, Gao W, Chen QF, Ramalingam S, Chye ML.

School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong
Kong, China.

In Arabidopsis thaliana, a family of six genes encodes acyl-CoA-binding proteins
(ACBPs) that show conservation at the acyl-CoA-binding domain. They are the
membrane-associated ACBP1 and ACBP2, extracellularly targeted ACBP3,
kelch-motif-containing ACBP4 and ACBP5, and 10-kDa ACBP6. The acyl-CoA domain in
each of ACBP1 to ACBP6 binds long-chain acyl-CoA esters in vitro, suggestive of
possible roles in plant lipid metabolism. We addressed here the use of
Arabidopsis ACBPs in conferring lead [Pb(II)] tolerance in transgenic plants
because the 10-kDa human ACBP has been identified as a molecular target for
Pb(II) in vivo. We investigated the effect of Pb(II) stress on the expression of
genes encoding Arabidopsis ACBP1, ACBP2 and ACBP6. We showed that the expression
of ACBP1 and ACBP2, but not ACBP6, in root is induced by Pb(II) nitrate
treatment. In vitro Pb(II)-binding assays indicated that ACBP1 binds Pb(II)
comparatively better, and ACBP1 was therefore selected for further
investigations. When grown on Pb(II)-containing medium, transgenic Arabidopsis
lines overexpressing ACBP1 were more tolerant to Pb(II)-induced stress than the
wild type. Accumulation of Pb(II) in shoots of the ACBP1-overepxressing plants
was significantly higher than wild type. The acbp1 mutant showed enhanced
sensitivity to Pb(II) when germinated and grown in the presence of Pb(II) nitrate
and tolerance was restored upon complementation using an ACBP1 cDNA. Our results
suggest that ACBP1 is involved in mediating Pb(II) tolerance in Arabidopsis with
accumulation of Pb(II) in shoots. Such observations of Pb(II) accumulation,
rather than Pb(II) extrusion, in the ACBP1-overexpressing plants implicate
possible use of ACBP1 in Pb(II) phytoremediation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18182029 [PubMed - indexed for MEDLINE]

118: Plant J. 2008 Apr;54(1):93-105. Epub 2008 Jan 7.

Rice SVP-group MADS-box proteins, OsMADS22 and OsMADS55, are negative regulators
of brassinosteroid responses.

Lee S, Choi SC, An G.

Department of Life Science and National Research Laboratory of Plant Functional
Genomics, Pohang University of Science and Technology (POSTECH), Pohang 790-784,
Korea.

Most short vegetative phase (SVP)-group MADS-box genes control meristem identity
and flowering time. Among the three SVP-group genes in rice, OsMADS47 has been
reported as a negative regulator of brassinosteroid (BR) responses. Here, we
investigated the functional roles of two close homologs, OsMADS22 and OsMADS55,
by generating single, double and triple RNAi lines and overexpression lines.
Analyses of the plants showed that their roles in regulating meristem identity
are well conserved; however, the involvement of these genes in determining
flowering time has diversified. Most importantly, OsMADS55 works as a major
negative regulator of BR responses, and OsMADS22 functions to support OsMADS55.
Whereas single OsMADS55 RNAi plants display weak BR responses in the lamina joint
(LJ), OsMADS22-OsMADS55 double and OsMADS22-OsMADS47-OsMADS55 triple RNAi plants
manifest dramatic BR responses with regard to LJ inclination, coleoptile
elongation and senescence. Stem elongation is also notably reduced in the double
and triple RNAi plants, probably because of BR oversensitivity. Expression
analyses indicate the diversified roles in age-dependent BR responses.
Altogether, our study demonstrates that all three rice SVP-group genes work as
negative regulators of BR responses, but that their spatial and temporal roles
are diversified.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18182025 [PubMed - indexed for MEDLINE]

119: Plant Mol Biol. 2008 Mar;66(5):491-502. Epub 2008 Jan 6.

DH1, a LOB domain-like protein required for glume formation in rice.

Li A, Zhang Y, Wu X, Tang W, Wu R, Dai Z, Liu G, Zhang H, Wu C, Chen G, Pan X.

Yangzhou University, Yangzhou, Jiangsu, 225009, People's Republic of China.

T-DNA tagging is a high throughput strategy for identifying and cloning
functional genes in plants. In this study, we screened 4416 lab-created T(1) rice
T-DNA tagged lines and identified a mutant, designated dh1 (degenerated hull1),
with phenotype of degenerated hull and naked pistils and stamens. Approximately
60% florets on the dh1 panicle defected in forming normal palea and lemma.
Instead, they formed degenerative velum-like or filamentous organs accompanying
with the lack of lodicules, stamens and pistils at different degree. A 361 bp of
genomic sequence flanking the T-DNA isolated using TAIL-PCR (Thermal asymmetric
interlaced PCR) co-segregated with the mutation phenotype. Results of blastn and
gene prediction revealed the T-DNA inserted into the promoter region of a
function-predicted gene at 283 bp upstream of its transcription start site (TSS).
The predicted gene encoded a LOB (Lateral Organ Boundaries) domain-like protein.
RT-PCR analyses indicated the transcription level of target candidate gene, DH1,
decreased significantly in dh1 mutant. RNAi aimed at DH1 in wild type plants
could partially result in the mutation phenotype of dh1. DH1 could also rescue
the mutation phenotype in the complement experiment. The result of transformation
by a fused expression vector, pDH1::GFP, revealed that DH1 had the keen spatial
and temporal characteristics of expressing at axillary bud, young panicle and
floral organs but not at root, leaf, node and culm, and strongly expressing at
young tissues but weakly at mature organs. The dh1 presented severer mutation
phenotype under relatively longer daylight than under shorter daylight implied
that shorter daylight induced the expression of gene(s) redundant to DH1 in
function and partially compensated for the loss-of-function. It is the first time
to report the LOB-domain gene participating in the development of floral organs
in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18180880 [PubMed - indexed for MEDLINE]

120: New Phytol. 2008;177(4):977-89. Epub 2007 Dec 19.

Capsicum annuum WRKY protein CaWRKY1 is a negative regulator of pathogen defense.

Oh SK, Baek KH, Park JM, Yi SY, Yu SH, Kamoun S, Choi D.

Plant Genome Research Center, KRIBB, P.O. Box 115, Yusung, Daejeon 305-600,
Korea.

Plants respond to pathogens by regulating a network of signaling pathways that
fine-tune transcriptional activation of defense-related genes. The aim of this
study was to determine the role of Capsicum annuum WRKY zinc finger-domain
transcription factor 1 (CaWRKY1) in defense. In previous studies, CaWRKY1 was
found to be rapidly induced in C. annuum (chili pepper) leaves by incompatible
and compatible pathogen inoculations, but the complexity of the network of the
WRKY family prevented the function of CaWRKY1 in defense from being elucidated.
Virus-induced gene silencing of CaWRKY1 in chili pepper leaves resulted in
decreased growth of Xanthomonas axonopodis pv. vesicatoria race 1.
CaWRKY1-overexpressing transgenic plants showed accelerated hypersensitive cell
death in response to infection with tobacco mosaic virus and Pseudomonas syringe
pv. tabaci. Lower levels of pathogenesis-related gene induction were observed in
CaWRKY1-overexpressing transgenic plants following salicylic acid (SA)
treatments. This work suggests that the newly characterized CaWRKY1, which is
strongly induced by pathogen infections and the signal molecule SA, acts as a
regulator to turn off systemic acquired resistance once the pathogen challenge
has diminished and to prevent spurious activation of defense responses at
suboptimal concentrations of SA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18179600 [PubMed - indexed for MEDLINE]

121: J Environ Qual. 2008 Jan 4;37(1):182-5. Print 2008 Jan-Feb.

Isotopic discrimination as a tool for organic farming certification in sweet
pepper.

del Amor FM, Navarro J, Aparicio PM.

Instituto Murciano de Investigaciö¨n y Desarrollo Agrario y Alimentario, 30150 La
Alberca, Murcia, Spain. franciscom.delamor@carm.es

Organic farming is a form of agriculture that excludes the use of synthetic
fertilizers, pesticides, and genetically modified organisms. These fertilizers
have been traditionally overused in conventional farming to avoid lost revenue,
but this often not does not take into account the potential contamination of
aquifers and river due to nitrate leaching. Transition to organic farming
practices could provide an instrument to reduce contamination and increase
potential income. It is difficult to determine to what extent those fertilizers
could have been used within a complete traceability of the production process. In
this experiment, we evaluated the use of (15)N/(14)N isotopic discrimination in
sweet pepper plants to test the hypothesis that synthetic fertilizers
significantly reduce (15)N/(14)N compared with exclusively organic practices.
Therefore, three common types of organic manures (sheep, hen, or horse) were
applied at a rate of 8 kg m(-2) with or without synthetic fertilizer amendments
under fully controlled environmental and irrigation conditions. Results indicate
that (i) use of synthetic fertilizers significantly reduced (15/14)N(2)vsN(2)atm
compared with treatments that only received water; (ii) with respect to the plant
organs, old leaves and fruits were more sensitive to the synthetic fertilizer
additions with reductions in (15/14)N(2)vsN(2)atm of 24.1 and 27.8%,
respectively; and (iii) independently of the organic manure used, no additional
fertilization (synthetic or organic) is required before 106 days after
transplanting at that dosage because plant fresh weight was not reduced.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18178891 [PubMed - indexed for MEDLINE]

122: J Microbiol Biotechnol. 2007 Dec;17(12):1944-8.

Effects of silkworm hemolymph on cell viability and hCTLA4Ig production in
transgenic rice cell suspension cultures.

Cheon SH, Lee KH, Kwon JY, Ryu HN, Yu DH, Choi YS, Kim DI.

Department of Biological Engineering, Inha University, Incheon 402-751, Korea.

Silkworm hemolymph (SH), prepared from fifth-instar larvae of Bombyx mori and
heat-treated at 60 degrees C for 30 min, was used to improve cell viability and
the production of human cytotoxic T-lymphocyte antigen 4-immunoglobulin
(hCTLA4Ig) in transgenic Oryza sativa L. cell suspension cultures. Even though SH
could not elevate cell viability at the concentrations up to 3% (v/v), addition
of 0.3% (v/v) SH to a culture medium enhanced the production of hCTLA4Ig by 36.8%
over an SH-free medium. Moreover, the production period of hCTLA4Ig could be
shortened in a 0.3% (v/v) SHadded medium compared with that in an SH-free
culture. As a result, addition of 0.3% (v/v) SH improved the productivity of
hCTLA4Ig significantly in transgenic rice cell cultures.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18167440 [PubMed - indexed for MEDLINE]

123: Nat Rev Genet. 2008 Feb;9(2):91-101.

Towards a better bowl of rice: assigning function to tens of thousands of rice
genes.

Jung KH, An G, Ronald PC.

Department of Plant Pathology, 1 Shields Avenue, UC Davis, Davis, California
95616, USA.

Rice, one of the most important food crops for humans, is the first crop plant to
have its genome sequenced. Rice whole-genome microarrays, genome tiling arrays
and genome-wide gene-indexed mutant collections have recently been generated.
With the availability of these resources, discovering the function of the
estimated 41,000 rice genes is now within reach. Such discoveries have broad
practical implications for understanding the biological processes of rice and
other economically important grasses such as cereals and bioenergy crops.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 18160965 [PubMed - indexed for MEDLINE]

124: Cytogenet Genome Res. 2007;119(1-2):147-53. Epub 2007 Dec 14.

Molecular characterisation and chromosomal localisation of a telomere-like
repetitive DNA sequence highly enriched in the C genome of Brassica.

Galvö¸o Bezerra dos Santos K, Becker HC, Ecke W, Bellin U.

Department of Crop Sciences, Georg-August-Universitö¤t Gö¶ttingen, Germany.
ksantos@gwdg.de

The aim of this work was to find C genome specific repetitive DNA sequences able
to differentiate the homeologous A (B. rapa) and C (B. oleracea) genomes of
Brassica, in order to assist in the physical identification of B. napus
chromosomes. A repetitive sequence (pBo1.6) highly enriched in the C genome of
Brassica was cloned from B. oleracea and its chromosomal organisation was
investigated through fluorescent in situ hybridisation (FISH) in B. oleracea (2n
= 18, CC), B. rapa (2n = 20, AA) and B. napus (2n = 38, AACC) genomes. The
sequence was 203 bp long with a GC content of 48.3%. It showed up to 89% sequence
identity with telomere-like DNA from many plant species. This repeat was clearly
underrepresented in the A genome and the in situ hybridisation showed its B.
oleracea specificity at the chromosomal level. Sequence pBo1.6 was localised at
interstitial and/or telomeric/subtelomeric regions of all chromosomes from B.
oleracea, whereas in B. rapa no signal was detected in most of the cells. In B.
napus 18 to 24 chromosomes hybridised with pBo1.6. The discovery of a sequence
highly enriched in the C genome of Brassica opens the opportunity for detailed
studies regarding the subsequent evolution of DNA sequences in polyploid genomes.
Moreover, pBo1.6 may be useful for the determination of the chromosomal location
of transgenic DNA in genetically modified oilseed rape. Copyright (c) 2007 S.
Karger AG, Basel.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18160795 [PubMed - indexed for MEDLINE]

125: Hypertension. 2008 Feb;51(2):540-6. Epub 2007 Dec 24.

Dietary n-3 polyunsaturated fatty acids and direct renin inhibition improve
electrical remodeling in a model of high human renin hypertension.

Fischer R, Dechend R, Qadri F, Markovic M, Feldt S, Herse F, Park JK, Gapelyuk A,
Schwarz I, Zacharzowsky UB, Plehm R, Safak E, Heuser A, Schirdewan A, Luft FC,
Schunck WH, Muller DN.

Medical Faculty of the Charitö©, Experimental and Clinical Research Center, Franz
Volhard Clinic and HELIOS Klinikum Berlin-Buch, Berlin, Germany.
robert.fischer@charite.de

We compared the effect n-3 polyunsaturated fatty acids (PUFAs) with direct renin
inhibition on electrophysiological remodeling in angiotensin II-induced cardiac
injury. We treated double-transgenic rats expressing the human renin and
angiotensinogen genes (dTGRs) from week 4 to 7 with n-3 PUFA ethyl-esters
(Omacor; 25-g/kg diet) or a direct renin inhibitor (aliskiren; 3 mg/kg per day).
Sprague-Dawley rats were controls. We performed electrocardiographic,
magnetocardiographic, and programmed electrical stimulation. Dietary n-3 PUFAs
increased the cardiac content of eicosapentaenoic and docosahexaenoic acid. At
week 7, mortality in dTGRs was 31%, whereas none of the n-3 PUFA- or
aliskiren-treated dTGRs died. Systolic blood pressure was modestly reduced in n-3
PUFA-treated (180+/-3 mm Hg) compared with dTGRs (208+/-5 mm Hg).
Aliskiren-treated dTGRs and Sprague-Dawley rats were normotensive (110+/-3 and
119+/-6 mm Hg, respectively). Both n-3 PUFA-treated and untreated dTGRs showed
cardiac hypertrophy and increased atrial natriuretic peptide levels. Prolonged
QRS and QT(c) intervals and increased T-wave dispersion in dTGRs were reduced by
n-3 PUFAs or aliskiren. Both treatments reduced arrhythmia induction from 75% in
dTGRs to 17% versus 0% in Sprague-Dawley rats. Macrophage infiltration and
fibrosis were reduced by n-3 PUFAs and aliskiren. Connexin 43, a mediator of
intermyocyte conduction, was redistributed to the lateral cell membranes in
dTGRs. n-3 PUFAs and aliskiren restored normal localization to the intercalated
disks. Thus, n-3 PUFAs and aliskiren improved electrical remodeling, arrhythmia
induction, and connexin 43 expression, despite a 70-mm Hg difference in blood
pressure and the development of cardiac hypertrophy.

PMID: 18158339 [PubMed - indexed for MEDLINE]

126: Anal Chim Acta. 2008 Jan 21;607(1):106-13. Epub 2007 Nov 19.

Development and validation of a sensitive enzyme immunoassay for surveillance of
Cry1Ab toxin in bovine blood plasma of cows fed Bt-maize (MON810).

Paul V, Steinke K, Meyer HH.

Physiology Weihenstephan, Technical University Munich, Weihenstephaner Berg 3,
85350 Freising, Germany. Vijay@wzw.tum.de

The increasing global adoption of genetically modified (GM) plant derivatives in
animal feed has provoked a strong demand for an appropriate detection method to
evaluate the existence of transgenic protein in animal tissues and animal
by-products derived from GM plant fed animals. A highly specific and sensitive
sandwich enzyme immunoassay for the surveillance of transgenic Cry1Ab protein
from Bt-maize in the blood plasma of cows fed on Bt-maize was developed and
validated according to the criteria of EU-Decision 2002/657/EC. The sandwich
assay is based on immuno-affinity purified polyclonal antibody raised against
Cry1Ab protein in rabbits. Native and biotinylated forms of this antibody served
as capture antibody and detection antibody for the ELISA, respectively.
Streptavidin-horseradish peroxidase conjugate and TMB substrate provided the
means for enzymatic colour development. The immunoassay allowed Cry1Ab protein
determination in bovine blood plasma in an analytical range of 0.4-100 ng mL(-1)
with a decision limit (CCalpha) of 1.5 ng mL(-1) and detection capability
(CCbeta) of 2.3 ng mL(-1). Recoveries ranged from 89 to 106% (mean value of 98%)
in spiked plasma. In total, 20 plasma samples from cows (n=7) fed non-transgenic
maize and 24 samples from cows (n=8) fed transgenic maize (collected before and,
after 1 and 2 months of feeding) were investigated for the presence of the Cry1Ab
protein. There was no difference amongst both groups (all the samples were below
1.5 ng mL(-1); CCalpha). No plasma sample was positive for the presence of the
Cry1Ab protein at CCalpha and CCbeta of the assay.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 18155416 [PubMed - indexed for MEDLINE]

127: BMC Plant Biol. 2007 Dec 20;7:67.

TaMSH7: a cereal mismatch repair gene that affects fertility in transgenic barley
(Hordeum vulgare L.).

Lloyd AH, Milligan AS, Langridge P, Able JA.

School of Agriculture, Food & Wine, The University of Adelaide, Waite Campus,
PMB1, Glen Osmond, South Australia, 5064, Australia. andrew.lloyd@adelaide.edu.au

BACKGROUND: Chromosome pairing, recombination and DNA repair are essential
processes during meiosis in sexually reproducing organisms. Investigating the
bread wheat (Triticum aestivum L.) Ph2 (Pairing homoeologous) locus has
identified numerous candidate genes that may have a role in controlling such
processes, including TaMSH7, a plant specific member of the DNA mismatch repair
family. RESULTS: Sequencing of the three MSH7 genes, located on the short arms of
wheat chromosomes 3A, 3B and 3D, has revealed no significant sequence divergence
at the amino acid level suggesting conservation of function across the
homoeogroups. Functional analysis of MSH7 through the use of RNAi
loss-of-function transgenics was undertaken in diploid barley (Hordeum vulgare
L.). Quantitative real-time PCR revealed several T0 lines with reduced MSH7
expression. Positive segregants from two T1 lines studied in detail showed
reduced MSH7 expression when compared to transformed controls and null
segregants. Expression of MSH6, another member of the mismatch repair family
which is most closely related to the MSH7 gene, was not significantly reduced in
these lines. In both T1 lines, reduced seed set in positive segregants was
observed. CONCLUSION: Results presented here indicate, for the first time, a
distinct functional role for MSH7 in vivo and show that expression of this gene
is necessary for wild-type levels of fertility. These observations suggest that
MSH7 has an important function during meiosis and as such remains a candidate for
Ph2.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18096080 [PubMed - indexed for MEDLINE]

128: FEMS Microbiol Ecol. 2008 Feb;63(2):156-68. Epub 2007 Dec 15.

Use of a novel nonantibiotic triple marker gene cassette to monitor high survival
of Pseudomonas fluorescens SBW25 on winter wheat in the field.

Jö¤derlund L, Hellman M, Sundh I, Bailey MJ, Jansson JK.

Department of Microbiology, Swedish University of Agricultural Sciences, Uppsala,
Sweden.

Pseudomonas fluorescens SBW25 was tagged with a triple marker gene cassette
containing gfp, encoding green fluorescent protein; luxAB, encoding luciferase;
and telABkilA, encoding tellurite resistance, and the tagged strain was monitored
in the first Swedish field release of a genetically modified microorganism (GMM).
The cells were inoculated onto winter wheat seeds and the GMM cells (SBW25:tgl)
were monitored in the field from September 2005 to May 2006 using plating,
luminometry and microscopic analyses. Cell numbers were high on all sampling
occasions and metabolically active cells were detected on all plant parts. Field
results were similar to those obtained in a parallel phytotron study, although
the amount of SBW25:tgl detected on shoots was significantly higher in the
phytotron than in the field. After winter, cell counts were 100-fold higher on
the roots and root-associated soil compared with prewinter measurements, although
the cells had a lower relative metabolic activity. The wheat seeds were naturally
infested with Microdochium nivale, but no treatment resulted in reduction of
disease symptoms. No SWB25:tgl cells were ever found in bulk soil or uninoculated
plants. The Swedish field trial results complement and contrast with prior field
studies performed with the same parent organism in the United Kingdom under
different soil, plant and climatic conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18093144 [PubMed - indexed for MEDLINE]

129: Environ Toxicol Chem. 2008 Jan;27(1):188-95.

Toxicological safety assessment of genetically modified Bacillus thuringiensis
with additional N-acyl homoserine lactonase gene.

Peng D, Zhou C, Chen S, Ruan L, Yu Z, Sun M.

State Key Laboratory of Agricultural Microbiology, College of Life Sciences and
Technology, Huazhong Agricultural University, Wuhan, People's Republic of China.

The aim of the present study is to evaluate the toxicology safety to mammals of a
genetically modified (GM) Bacillus thuringiensis with an additional N-acyl
homoserine lactones gene (aiiA), which possesses insecticidal activity together
with restraint of bacterial pathogenicity and is intended for use as a
multifunctional biopesticide. Safety assessments included an acute oral toxicity
test and 28-d animal feeding study in Wistar rats, primary eye and dermal
irritation in Zealand White rabbits, and delayed contact hypersensitivity in
guinea pigs. Tests were conducted using spray-dried powder preparation. This GM
product showed toxicity neither in oral acute toxicity test nor in 28-d animal
feeding test at a dose of 5,000 mg/kg body weight. During the animal feeding
test, there were no significant differences in growth, food and water
consumption, hematology, blood biochemical indices, organ weights, and
histopathology finding between rats in controls and tested groups. Tested animals
in primary eye and dermal irritation and delayed contact hypersensitivity test
were also devoid of any toxicity compared to controls. All the above results
demonstrated that the GM based multifunctional B. thuringiensis has low toxicity
and low eye and dermal irritation and would not cause hypersensitivity to
laboratory mammals and therefore could be regarded as safe for use as a
pesticide.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18092859 [PubMed - indexed for MEDLINE]

130: Mol Syst Biol. 2007;3:158. Epub 2007 Dec 18.

Synthetic biology: promises and challenges.

Serrano L.

EMBL-CRG Systems Biology Unit, Centro de Regulaciö¨n Genomica CRG, Barcelona,
Spain.

Publication Types:
Editorial

PMID: 18091727 [PubMed - indexed for MEDLINE]

131: Plant J. 2008 Apr;54(1):43-55. Epub 2007 Dec 15.

Partial demethylation of oligogalacturonides by pectin methyl esterase 1 is
required for eliciting defence responses in wild strawberry (Fragaria vesca).

Osorio S, Castillejo C, Quesada MA, Medina-Escobar N, Brownsey GJ, Suau R,
Heredia A, Botella MA, Valpuesta V.

Departamento de Biologö­a Molecular y Bioquö­mica, Universidad de Mö¡laga, 29071
Mö¡laga, Spain.

In addition to the role of the cell wall as a physical barrier against pathogens,
some of its constituents, such as pectin-derived oligogalacturonides (OGA), are
essential components for elicitation of defence responses. To investigate how
modifications of pectin alter defence responses, we expressed the fruit-specific
Fragaria x ananassa pectin methyl esterase FaPE1 in the wild strawberry Fragaria
vesca. Pectin from transgenic ripe fruits differed from the wild-type with regard
to the degree and pattern of methyl esterification, as well as the average size
of pectin polymers. Purified oligogalacturonides from the transgenic fruits
showed a reduced degree of esterification compared to oligogalacturonides from
wild-type fruits. This reduced esterification is necessary to elicit defence
responses in strawberry. The transgenic F. vesca lines had constitutively
activated pathogen defence responses, resulting in higher resistance to the
necrotropic fungus Botrytis cinerea. Further studies in F. vesca and Nicotiana
benthamiana leaves showed that the elicitation capacity of the
oligogalacturonides is more specific than previously envisaged.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18088306 [PubMed - indexed for MEDLINE]

132: Plant Biotechnol J. 2008 Apr;6(3):295-300. Epub 2007 Dec 11.

Fruit-specific suppression of the ethylene receptor LeETR4 results in
early-ripening tomato fruit.

Kevany BM, Taylor MG, Klee HJ.

Plant Molecular and Cellular Biology Program, Horticultural Sciences, University
of Florida, Gainesville, FL 32611-0690, USA.

Tomato is an economically important crop and a significant dietary source of
important phytochemicals, such as carotenoids and flavonoids. Although it has
been known for many years that the plant hormone ethylene is essential for the
ripening of climacteric fruits, its role in fruit growth and maturation is much
less well understood. In this study, data are presented which indicate that
fruit-specific suppression of the ethylene receptor LeETR4 causes early ripening,
whereas fruit size, yield and flavour-related chemical composition are largely
unchanged. Early fruit ripening is a highly desirable and valuable trait, and the
approach demonstrated here should be applicable to any fruit species requiring
ethylene to ripen. These results demonstrate that ethylene receptors probably act
as biological clocks regulating the onset of tomato fruit ripening.

PMID: 18086233 [PubMed - indexed for MEDLINE]

133: Plant Biotechnol J. 2008 Apr;6(3):281-94. Epub 2007 Dec 10.

Higher biomass accumulation by increasing phosphoribosylpyrophosphate synthetase
activity in Arabidopsis thaliana and Nicotiana tabacum.

Koslowsky S, Riegler H, Bergmö¼ller E, Zrenner R.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie, Am Mö¼hlenberg 1, 14476
Potsdam Golm, Germany.

Plants are able to produce all the organic compounds required for development and
growth. As developmental processes and metabolic pathways use a common resource
pool, the tight regulation of the distribution of metabolites between growth,
production of defence compounds and storage products can be assumed. A transgenic
approach was used to investigate the importance of supplying the key intermediate
phosphoribosylpyrophosphate (PRPP) for plant growth and biomass accumulation in
the model plant Arabidopsis thaliana and in Nicotiana tabacum. For this purpose,
the Ashbya gossypii genes coding for either PRPP synthetase (PRS) or a mutated
variant of the same gene were over-expressed under the control of a constitutive
promoter. It was shown that increased PRS activity in A. thaliana or N. tabacum
leads to a substantial increase in biomass accumulation under different
standardized growth conditions. Growth enhancement was accompanied by significant
changes in the amount of sugars and other metabolites. This study provides
evidence that the supply of PRPP co-limits growth rates, and has obvious
implications for biotechnological strategies aiming to increase plant biomass as
an alternative renewable energy source.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18086232 [PubMed - indexed for MEDLINE]

134: Toxicol Sci. 2008 Apr;102(2):425-32. Epub 2007 Dec 15.

A gene-shuffled glyphosate acetyltransferase protein from Bacillus licheniformis
(GAT4601) shows no evidence of allergenicity or toxicity.

Delaney B, Zhang J, Carlson G, Schmidt J, Stagg B, Comstock B, Babb A, Finlay C,
Cressman RF, Ladics G, Cogburn A, Siehl D, Bardina L, Sampson H, Han Y.

Pioneer Hi-Bred International, Inc., Johnston, Iowa 50131, USA.

The glyphosate acetyltransferase (gat) gene from Bacillus licheniformis was
subjected to multiple rounds of gene shuffling to optimize kinetics of
corresponding GAT proteins to acetylate the herbicide active ingredient
glyphosate. Genetically modified soybeans expressing the gat4601 gene (356043
soybeans) are tolerant to the application of glyphosate. The current manuscript
reports the outcome of the allergenicity and toxicity assessment for the GAT4601
protein. Bioinformatic comparison of the amino acid sequence of GAT4601 did not
identify similarities to known allergenic or toxic proteins. In vitro studies
conducted with heterologously produced GAT4601 protein demonstrated that it was
rapidly degraded in simulated gastric fluid containing pepsin (< 30 s) and in
simulated intestinal fluid containing pancreatin (< 2 min) and completely
inactivated at temperatures above 56 degrees C. The GAT4601 protein expressed in
planta is not glycosylated and similar protein profiles were observed in flour
extracts from 356043 soybeans and nontransgenic near isoline comparator soybeans
(Jack) using serum from soy allergic persons. No evidence of adverse effects was
observed in mice following acute oral exposure to 2000 mg/kg of GAT4601 protein
or in a repeated dose dietary exposure study at doses of 800-1000 mg/kg/day. This
comprehensive assessment demonstrates that the GAT4601 protein does not present a
risk for adverse effects in humans when used in the context of agricultural
biotechnology.

Publication Types:
In Vitro
Research Support, Non-U.S. Gov't

PMID: 18084044 [PubMed - indexed for MEDLINE]

135: Trends Plant Sci. 2008 Jan;13(1):28-35. Epub 2007 Dec 20.

Folate biofortification in food plants.

Bekaert S, Storozhenko S, Mehrshahi P, Bennett MJ, Lambert W, Gregory JF 3rd,
Schubert K, Hugenholtz J, Van Der Straeten D, Hanson AD.

Department of Molecular Genetics, Ghent University, K.L. Ledeganckstraat 35,
B-9000 Gent, Belgium.

Folate deficiency is a global health problem affecting many people in the
developing and developed world. Current interventions (industrial food
fortification and supplementation by folic acid pills) are effective if they can
be used but might not be possible in less developed countries. Recent advances
demonstrate that folate biofortification of food crops is now a feasible
complementary strategy to fight folate deficiency worldwide. The genes and
enzymes of folate synthesis are sufficiently understood to enable metabolic
engineering of the pathway, and results from pilot engineering studies in plants
(and bacteria) are encouraging. Here, we review the current status of
investigations in the field of folate enhancement on the eve of a new era in food
fortification.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 18083061 [PubMed - indexed for MEDLINE]

136: Acta Virol. 2007;51(3):157-62.

Expression of the barley yellow dwarf virus-GAV movement protein and its
detection in the infected and transgenic plants.

Xie J, Wang X, Liu Y, Peng Y, Zhou G.

Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing,
China.

Movement proteins (MPs) that facilitate virus movement in the plants were
identified in a number of plant viruses. In this study, full-length MP gene of
the Chinese isolate Barley yellow dwarf virus-GAV (BYDV-GAV) was cloned and
expressed in Escherichia coli. About 32% of the expressed MP was soluble
providing the concentration of isopropyl-beta-D-galactopyranoside (IPTG), time of
the induction, temperature and shaking speed were optimized. The soluble MP was
purified using nickel-affinity column. Immune serum prepared against purified MP
was used for the detection of MP in the BYDV-GAV infected leaves of oat and in
the leaves of transgenic wheat plants expressing the full-length and truncated MP
gene.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18076305 [PubMed - indexed for MEDLINE]

137: Nature. 2007 Dec 13;450(7172):928-9.

Showdown for Europe.

Abbott A, Schiermeier Q.

Publication Types:
News

PMID: 18075535 [PubMed - indexed for MEDLINE]

138: Nature. 2007 Dec 13;450(7172):921.

Directive action required.

[No authors listed]

Publication Types:
Editorial

PMID: 18075528 [PubMed - indexed for MEDLINE]

139: Biotechniques. 2007 Nov;43(5):649-50, 652, 654 passim.

High-efficiency thermal asymmetric interlaced PCR for amplification of unknown
flanking sequences.

Liu YG, Chen Y.

Key Laboratory of Plant Functional Genomics and Biotechnology of Education
Department, College of Life Sciences, South China Agricultural University,
Guangzhou, China. ygliuy@scau.edu.cn

Isolation of unknown DNA sequences flanked by known sequences is an important
task in molecular biology research. Thermal asymmetric interlaced PCR (TAIL-PCR)
is an effective method for this purpose. However the success rate of the original
TAIL-PCR needs to be increased, and it is more desirable to obtain target
products with larger sizes. Here we present a substantially improved TAIL-PCR
procedure with special primer design and optimized thermal conditions. This
high-efficiency TAIL-PCR (hiTAIL-PCR) combines the advantages of the TAIL-cycling
and suppression-PCR, thus it can block the amplification of nontarget products
and suppress small target ones, but allow efficient amplification of large target
sequences. Using this method, we isolated genomic flanking sequences of T-DNA
insertions from transgenic rice lines. In our tests, the success rates of the
reactions were higher than 90%, and in most cases the obtained major products had
sizes of 1-3 kb.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18072594 [PubMed - indexed for MEDLINE]

140: Cell Res. 2008 Apr;18(4):508-21.

Constitutive expression of pathogen-inducible OsWRKY31 enhances disease
resistance and affects root growth and auxin response in transgenic rice plants.

Zhang J, Peng Y, Guo Z.

State Key Laboratory of Agrobiotechnology, Department of Plant Pathology, China
Agricultural University, Yuanmingyuan West Road 2, Beijing 100094, China.

WRKY transcription factors have many regulatory roles in response to biotic and
abiotic stresses. In this study, we isolated a rice WRKY gene (OsWRKY31) that is
induced by the rice blast fungus Magnaporthe grisea and auxin. This gene encodes
a polypeptide of 211 amino-acid residues and belongs to a subgroup of the rice
WRKY gene family that probably originated after the divergence of monocot and
dicot plants. OsWRKY31 was found to be localized to the nucleus of onion
epidermis cells to transiently express OsWRKY31-eGFP fusion protein. Analysis of
OsWRKY31 and its mutants fused with a Gal4 DNA-binding domain indicated that
OsWRKY31 has transactivation activity in yeast. Overexpression of the OsWRKY31
gene was found to enhance resistance against infection with M. grisea, and the
transgenic lines exhibited reduced lateral root formation and elongation compared
with wild-type and RNAi plants. The lines with overexpression showed constitutive
expression of many defense-related genes, such as PBZ1 and OsSci2, as well as
early auxin-response genes, such as OsIAA4 and OsCrl1 genes. Furthermore, the
plants with overexpression were less sensitive to exogenously supplied IBA, NAA
and 2,4-D at high concentrations, suggesting that overexpression of the OsWRKY31
gene might alter the auxin response or transport. These results also suggest that
OsWRKY31 might be a common component in the signal transduction pathways of the
auxin response and the defense response in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18071364 [PubMed - indexed for MEDLINE]

141: J Eukaryot Microbiol. 2007 Nov-Dec;54(6):465-7.

In vitro culture of the obligate parasite Spongospora subterranea (cercozoa;
plasmodiophorida) associated with root-inducing transferred-DNA transformed
potato hairy roots.

Qu X, Christ BJ.

Department of Plant Pathology, Pennsylvania State University, University Park,
Pennsylvania 16802, USA.

Spongospora subterranea is a soil-borne, obligate parasitic protist that causes
powdery scab of potatoes. In this study, an in vitro culture system was developed
for the maintenance and proliferation of the protist in potato hairy roots. The
hairy roots of potato were induced in vitro with Agrobacterium rhizogenes.
Cystosori of S. subterranea from potato scab lesions were surface disinfested and
used to inoculate potato hairy roots. Plasmodia, zoosporangia, and cystosori were
observed microscopically in the hairy roots within 6 wk after inoculation,
indicating the completion of the life cycle of S. subterranea in vitro. This is
the first in vitro culture system for S. subterranea, and will be a valuable tool
to study fundamental and practical aspects of the biology of the parasite.

Publication Types:
Evaluation Studies
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18070323 [PubMed - indexed for MEDLINE]

142: New Phytol. 2008;177(3):743-55. Epub 2007 Dec 7.

Isolate specificity of quantitative trait loci for partial resistance of barley
to Puccinia hordei confirmed in mapping populations and near-isogenic lines.

Marcel TC, Gorguet B, Ta MT, Kohutova Z, Vels A, Niks RE.

Laboratory of Plant Breeding, Graduate School for Experimental Plant Sciences,
Wageningen University, Droevendaalsesteeg 1, 6708 PB Wageningen, The Netherlands.

Partial resistance is considered race-nonspecific and durable, consistent with
the concept of 'horizontal' resistance. However, detailed observations of partial
resistance to leaf rust (Puccinia hordei) in barley (Hordeum vulgare) revealed
small cultivar x isolate interactions, suggesting a minor-gene-for-minor-gene
interaction model, similar to so-called 'vertical' resistance. Three consistent
quantitative trait loci (QTLs), labelled Rphq2, Rphq3 and Rphq4, that were
detected in the cross susceptible L94 x partially resistant Vada have been
incorporated into the L94 background to obtain near-isogenic lines (NILs). Three
isolates were used to map QTLs on seedlings of the L94 x Vada population and to
evaluate the effect of each QTL on adult plants of the respective NILs under
field conditions. Rphq2 had a strong effect in seedlings but almost no effect in
adult plants, while Rphq3 was effective in seedlings and in adult plants against
all three isolates. However, Rphq4 was effective in seedlings and in adult plants
against two isolates but ineffective in both development stages against the
third, demonstrating a clear and reproducible isolate-specific effect. The
resistance governed by the three QTLs was not associated with a hypersensitive
reaction. Those results confirm the minor-gene-for-minor-gene model suggesting
specific interactions between QTLs for partial resistance and P. hordei isolates.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18069952 [PubMed - indexed for MEDLINE]

143: J Agric Food Chem. 2008 Jan 9;56(1):272-80. Epub 2007 Dec 11.

Three polyphenol oxidases from red clover (Trifolium pratense) differ in
enzymatic activities and activation properties.

Schmitz GE, Sullivan ML, Hatfield RD.

US Dairy Forage Research Center, Agricultural Research Service, US Department of
Agriculture, Madison, WI 53706, USA.

Polyphenol oxidases (PPOs) oxidize o-diphenols to o-quinones, which cause
browning reactions in many wounded fruits, vegetables, and plants including the
forage crop red clover (Trifolium pratense L.). Production of o-quinones in red
clover inhibits postharvest proteolysis during the ensiling process. The cDNAs
encoding three red clover PPOs were expressed individually in alfalfa (Medicago
sativa L.), which lacks detectable endogenous foliar PPO activity and
o-diphenols. Several physical and biochemical characteristics of the red clover
PPOs in alfalfa extracts were determined. In transgenic alfalfa extracts, red
clover PPOs exist in a latent state and are activated (10-40-fold increase in
activity) by long incubations (>2 days) at ambient temperature or short
incubations (<10 min) at > or =65 degrees C. PPO1 appears to be more stable at
high temperatures than PPO2 or PPO3. During incubation at ambient temperature,
the molecular masses of the PPO enzymes were reduced by approximately 20 kDa. The
apparent pH optima of latent PPO1, PPO2, and PPO3 are 5.5, 6.9, and 5.1,
respectively, and latent PPO1 is slightly activated (~5-fold) by low pH.
Activation of the PPOs shifts the pH optima to approximately 7, and the activated
PPOs retain substantial levels of activity as the pH increases above their
optima. The latent and activated PPOs were surveyed for ability to oxidize
various o-diphenols, and activation of the PPOs had little effect on substrate
specificity. Activation increases the V max but not the affinity of the PPO
enzymes for caffeic acid. Results indicate red clover PPOs undergo structural and
kinetic changes during activation and provide new insights to their effects in
postharvest physiology.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 18069787 [PubMed - indexed for MEDLINE]

144: Z Naturforsch [C]. 2007 Sep-Oct;62(9-10):743-6.

Inheritance and expression of transgenes through anther culture of transgenic hot
pepper.

Kim YS, Kuk YI, Kim KM.

Kumho Life and Environmental Science Laboratory & Agricultural Plant Stress
Research Center, Chonnam National University, 300 Yongbong-dong, Buk-gu, Gwangju
500-757, Republic of Korea.

Anther cultures have been developed from transgenic donor peppers carrying
HPT/J1-1. Eight out of sixteen plants produced from an anther culture set pepper
fruits. Southern blot analysis of donors revealed two independent plants with a
single copy of the integrated transgene. PCR and RT-PCR results showed the
inheritance of HPT/J1-1 and expression of J1-1 in A1. All A1 progeny derived from
transgenic anthers had resistance to hygromycin. They grew normally and showed
similar phenotypes to the wild-type. Therefore, the use of an anther culture
system coupled with genetic transformation in breeding programs will greatly
facilitate the genetic improvement of pepper plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18069249 [PubMed - indexed for MEDLINE]

145: Nat Biotechnol. 2007 Dec;25(12):1356; author reply 1359-60.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

GM soybeans--revisiting a controversial format.

Cummins J.

Publication Types:
Comment
Letter

PMID: 18066023 [PubMed - indexed for MEDLINE]

146: Nat Biotechnol. 2007 Dec;25(12):1356-8.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

Response to GM soybeans-revisiting a controversial format.

[No authors listed]

Publication Types:
Comment

PMID: 18066022 [PubMed - indexed for MEDLINE]

147: Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

GM soybeans--revisiting a controversial format.

Ho MW, Saunders PT.

Publication Types:
Comment
Letter

PMID: 18066021 [PubMed - indexed for MEDLINE]

148: Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

GM soybeans--revisiting a controversial format.

Leifert C.

Publication Types:
Comment
Letter

PMID: 18066020 [PubMed - indexed for MEDLINE]

149: Nat Biotechnol. 2007 Dec;25(12):1355-6; author reply 1359-60.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

GM soybeans--revisiting a controversial format.

Heinemann JA, Traavik T.

Publication Types:
Comment
Letter

PMID: 18066019 [PubMed - indexed for MEDLINE]

150: Nat Biotechnol. 2007 Dec;25(12):1354-5; author reply 1359-60.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

GM soybeans--revisiting a controversial format.

John B.

Publication Types:
Comment
Letter

PMID: 18066018 [PubMed - indexed for MEDLINE]

151: Nat Biotechnol. 2007 Dec;25(12):1351-4; author reply 1359-60.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):981-7.

GM soybeans--revisiting a controversial format.

Ermakova IV.

Publication Types:
Comment
Letter

PMID: 18066017 [PubMed - indexed for MEDLINE]

152: Nat Biotechnol. 2007 Dec;25(12):1330.

Comment in:
Nat Biotechnol. 2008 Apr;26(4):379; discussion 379-80.

Another inconvenient truth. In Europe, no one apparently wants to listen if you
have good news about genetically modified organisms (GMOs).

[No authors listed]

Publication Types:
Editorial

PMID: 18066008 [PubMed - indexed for MEDLINE]

153: RNA. 2008 Feb;14(2):217-24. Epub 2007 Dec 7.

Faithful editing of a tomato-specific mRNA editing site in transgenic tobacco
chloroplasts.

Karcher D, Kahlau S, Bock R.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie, D-14476 Potsdam-Golm,
Germany.

RNA editing sites and their site-specific trans-acting recognition factors are
thought to have coevolved. Hence, evolutionary loss of an editing site by a
genomic mutation is normally followed by the loss of the specific recognition
factor for this site, due to the absence of selective pressure for its
maintenance. Here, we have tested this scenario for the only tomato-specific
plastid RNA editing site. A single C-to-U editing site in the tomato rps12 gene
is absent from the tobacco and nightshade plastid genomes, where the presence of
a genomic T nucleotide obviates the need for editing of the rps12 mRNA. We have
introduced the tomato editing site into the tobacco rps12 gene by plastid
transformation and find that, surprisingly, this heterologous site is efficiently
edited in the transplastomic plants. This suggests that the trans-acting
recognition factor for the rps12 editing site has been maintained, presumably
because it serves another function in tobacco plastids. Bioinformatics analyses
identified an editing site in the rpoB gene of tobacco and tomato whose sequence
context exhibits striking similarity to that of the tomato rps12 editing site.
This may suggest that requirement for rpoB editing resulted in maintenance of the
rps12 editing activity or, alternatively, the pre-existing rpoB editing activity
facilitated the evolution of a novel editing site in rps12.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18065714 [PubMed - indexed for MEDLINE]

154: Dokl Biochem Biophys. 2007 Sep-Oct;416:290-3.

Synthesis of hepatitis B virus surface antigen in tomato plants transgenic for
the preS2-S gene.

Salyaev K, Rekoslavskaya NI, Stolbikov AS, Hammond RW, Shchelkunov SN.

Siberian Institute of Plant Physiology and Biochernistry, Siberian Branch,
Russian Academy of Sciences, Irkutsk, 664033, Russia.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18064835 [PubMed - indexed for MEDLINE]

155: Biotechnol J. 2008 Mar;3(3):354-9.

Protoplast isolation and transient gene expression in switchgrass, Panicum
virgatum L.

Mazarei M, Al-Ahmad H, Rudis MR, Stewart CN Jr.

Department of Plant Sciences, University of Tennessee, Knoxville, TN 37996-4561,
USA.

Transient assay systems using protoplasts have been utilized in several plant
species and are a powerful tool for rapid functional gene analysis and
biochemical manipulations. A protoplast system has not been used in switchgrass
(Panicum virgatum L.), even though it is a bioenergy crop that has received
considerable attention. Here we report the first protocol to isolate large
numbers of viable protoplasts from both leaves and roots of two switchgrass
genotypes. Furthermore, we demonstrate transient expression of PEG-mediated DNA
uptake in the isolated protoplasts by measuring the activity of
beta-glucuronidase (GUS) reporter gene driven by either the Cauliflower mosaic
virus (CaMV) 35S promoter or the maize ubiquitin 1 promoter. Protoplast
transformation with either the 35S or the ubiquitin promoter resulted in an
increase in GUS activity compared to the untransformed controls; however, the
extent of GUS activity was considerably higher for the ubiquitin promoter than
for the 35S promoter. Collectively, our results indicate an efficient protoplast
isolation and transient assay system that can be used to facilitate gene
expression studies in switchgrass.

Publication Types:
Research Support, Non-U.S. Gov't
Technical Report

PMID: 18064611 [PubMed - indexed for MEDLINE]

156: Theor Appl Genet. 2008 Feb;116(4):455-63. Epub 2007 Dec 7.

Effects of defoliating insect resistance QTLs and a cry1Ac transgene in soybean
near-isogenic lines.

Zhu S, Walker DR, Boerma HR, All JN, Parrott WA.

Department of Crop and Soil Sciences, University of Georgia, Athens, GA 30602,
USA.

The crystal proteins coded by transgenes from Bacillus thuringiensis (Bt) have
shown considerable value in providing effective insect resistance in a number of
crop species, including soybean, Glycine max (L.) Merr. Additional sources of
soybean insect resistance would be desirable to manage the development of
tolerance/resistance to crystal proteins by defoliating insects and to sustain
the deployment of Bt crops. The objective of this study was to evaluate the
effects and interactions of three insect resistance quantitative trait loci
(QTLs; QTL-M, QTL-H, and QTL-G) originating from Japanese soybean PI 229358 and a
cry1Ac gene in a "Benning" genetic background. A set of 16 BC(6)F(2)-derived near
isogenic lines (NILs) was developed using marker-assisted backcrosses and
evaluated for resistance to soybean looper [SBL, Pseudoplusia includens (Walker)]
and corn earworm [CEW, Helicoverpa zea (Boddie)] in field cage, greenhouse, and
detached leaf assays. Both Bt and QTL-M had significantly reduced defoliation by
both SBL and CEW and reduced larval weight of CEW. The antibiosis QTL-G had a
significant effect on reducing CEW larval weight and also a significant effect on
reducing defoliation by SBL and CEW in some assays. The antixenosis QTL-H had no
main effect, but it appeared to function through interaction with QTL-M and
QTL-G. Adding QTL-H and QTL-G further enhanced the resistance of the Bt and QTL-M
combination to CEW in the field cage assay. These results should help guide the
development of strategies for effective management of insect pests and for
sustainable deployment of Bt genes.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18064435 [PubMed - indexed for MEDLINE]

157: Ying Yong Sheng Tai Xue Bao. 2007 Sep;18(9):2061-8.

[Effects of crop arrangement patterns on arthropod community structure in
transgenic bollworm-resistant cotton fields]

[Article in Chinese]

Guo JY, Wan FH, Hu YH, Yan Y.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of
Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094,
China. guojy406@163.com

The study on the arthropod community structures in transgenic bollworm-resistant
cotton (cv. Lumianyan 15) fields with different crop arrangement patterns showed
that compared with those in mono-cultured cotton plot, there were more species of
arthropod community and of phytophagous pest and natural enemy sub-communities as
well as more individuals of neutral arthropods in vegetable-, fruit tree-, and
peanut-cotton plots. More individuals of phytophagous pests in vegetable- and
peanut-cotton plots, and natural enemies in vegetable- and fruit tree-cotton
plots were observed. The arthropod community in peanut- and fruit tree-cotton
plots had the highest similarity, while that in mono-cultured cotton and
vegetable-cotton plots had the lowest one. The Renyi diversity index indicated
that compared with mono-cultured cotton plot, vegetable-cotton plot had lower
diversities of arthropod community and pest sub-community, while fruit tree- and
peanut-cotton plots had higher diversities of arthropod community and natural
enemy sub-community, and of arthropod community and pest sub-community diversity,
respectively. It was concluded that fruit tree-cotton was a recommendable crop
arrangement pattern for transgenic bollworm-resistant cotton.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 18062314 [PubMed - indexed for MEDLINE]

158: J Chromatogr A. 2008 Jan 4;1177(1):195-8. Epub 2007 Nov 19.

Analysis of glyphosate and aminomethylphosphonic acid by capillary
electrophoresis with electrochemiluminescence detection.

Chiu HY, Lin ZY, Tu HL, Whang CW.

Department of Chemistry, Tunghai University, Taichung 407, Taiwan.

A capillary electrophoresis (CE) method coupled with electrochemiluminescence
(ECL) detection for the analysis of glyphosate (GLY) and its major metabolite
aminomethylphosphonic acid (AMPA) is presented. Complete separation of GLY and
AMPA was achieved in 8 min using a background electrolyte of 20 mM sodium
phosphate (pH 9.0) and a separation voltage of 21 kV. ECL detection was performed
with an indium tin oxide (ITO) working electrode bias at 1.6 V (vs. a Pt-wire
reference) in a 30 0mM sodium phosphate buffer (pH 8.0) containing 3.5mM Ru(bpy)3
2+ (where bpy=2.2'-bipyridyl). Linear correlation (r>or=0.997) between ECL
intensity and analyte concentration was obtained in the ranges 0.169-16.9 and
5.55-111 microg ml(-1) for GLY and AMPA, respectively. The limits of detection
(LODs) for GLY and AMPA in water were 0.06 microg ml(-1) and 4.04 microg ml(-1),
respectively. The developed method was applied to the analysis of GLY in
soybeans. The LOD of GLY in soybean was 0.6 microg g(-1). Total analysis time
including sample pretreatment was less than 1h.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18061199 [PubMed - indexed for MEDLINE]

159: Mol Biotechnol. 2008 Jan;38(1):41-55.

Contributions of microorganisms to industrial biology.

Demain AL, Adrio JL.

Research Institute for Scientists Emeriti (R.I.S.E.), Drew University, Madison,
NJ 07940, USA. ademain@drew.edu

Life on earth is not possible without microorganisms. Microbes have contributed
to industrial science for over 100 years. They have given us diversity in
enzymatic content and metabolic pathways. The advent of recombinant DNA brought
many changes to industrial microbiology. New expression systems have been
developed, biosynthetic pathways have been modified by metabolic engineering to
give new metabolites, and directed evolution has provided enzymes with modified
selectability, improved catalytic activity and stability. More and more genomes
of industrial microorganisms are being sequenced giving valuable information
about the genetic and enzymatic makeup of these valuable forms of life. Major
tools such as functional genomics, proteomics, and metabolomics are being
exploited for the discovery of new valuable small molecules for medicine and
enzymes for catalysis.

Publication Types:
Review

PMID: 18060538 [PubMed - indexed for MEDLINE]

160: Biotechnol Adv. 2008 Mar-Apr;26(2):162-8. Epub 2007 Nov 12.

Advances in development of transgenic pulse crops.

Eapen S.

Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre,
Mumbai-400085, India. eapenhome@yahoo.com

It is three decades since the first transgenic pulse crop has been developed.
Todate, genetic transformation has been reported in all the major pulse crops
like Vigna species, Cicer arietinum, Cajanus cajan, Phaseolus spp, Lupinus spp,
Vicia spp and Pisum sativum, but transgenic pulse crops have not yet been
commercially released. Despite the crucial role played by pulse crops in tropical
agriculture, transgenic pulse crops have not moved out from laboratories to large
farm lands compared to their counterparts - 'cereals' and the closely related
leguminous oil crop - 'soybean'. The reason for lack of commercialization of
transgenic pulse crops can be attributed to the difficulty in developing
transgenics with reproducibility, which in turn is due to lack of competent
totipotent cells for transformation, long periods required for developing
transgenics and lack of coordinated research efforts by the scientific community
and long term funding. With optimization of various factors which influence
genetic transformation of pulse crops, it will be possible to develop transgenic
plants in this important group of crop species with more precision and
reproducibility. A translation of knowledge from information available in
genomics and functional genomics in model legumes like Medicago truncatula and
Lotus japonicus relating to factors which contribute to enhancing crop yield and
ameliorate the negative consequences of biotic and abiotic stress factors may
provide novel insights for genetic manipulation to improve the productivity of
pulse crops.

Publication Types:
Review

PMID: 18055156 [PubMed - indexed for MEDLINE]

161: Protein Expr Purif. 2008 Feb;57(2):127-35. Epub 2007 Nov 20.

Expression of active recombinant human lactoferrin in the milk of transgenic
goats.

Zhang J, Li L, Cai Y, Xu X, Chen J, Wu Y, Yu H, Yu G, Liu S, Zhang A, Chen J,
Cheng G.

Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai
200025, China.

This report details the establishment of a transgenic goat model in order to
produce human lactoferrin (hLf) in the mammary gland for large-scale application
and research. Two transgenic male goats were generated by microinjecting sequence
encoding hLf cDNA to the pronuclear. In the two lines, derived from the two
founders, eight lactating female goats could secrete recombinant human
lactoferrin (rhLf) at concentrations of up to 0.765 mg/ml. The method of
purifying the rhLf from the milk was achieved using ion-exchange chromatography
and resulted in 97% purity. Biochemical and physicochemical characteristics of
rhLf were similar to native lactoferrin (nhLf); this included N-terminal
sequence, isoelectric point, molecular mass, glycosylation,
iron-binding/releasing ability, thermal stability, and proteolysis. The rhLf
showed broad spectrum antibacterial activity inhibiting the growth of several
pathogenic bacterial strains. Also investigated, although to a lesser degree, was
a practicable pasteurization method for the downstream processing of rhLf and,
further, a method for the oral administration of rhLf. On the basis of these
results, our studies show an optimistic and promising approach for the
large-scale production and therapeutic application of rhLf expressed in
transgenic goats.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18054499 [PubMed - indexed for MEDLINE]

162: Mol Plant Microbe Interact. 2008 Jan;21(1):30-9.

Naturally occurring broad-spectrum powdery mildew resistance in a Central
American tomato accession is caused by loss of mlo function.

Bai Y, Pavan S, Zheng Z, Zappel NF, Reinstö¤dler A, Lotti C, De Giovanni C,
Ricciardi L, Lindhout P, Visser R, Theres K, Panstruga R.

Laboratory of Plant Breeding, Plant Sciences Group, Wageningen University,
Droevendaalsesteeg 1, Wageningen, The Netherlands. bai.yuling@wur.nl

The resistant cherry tomato (Solanum lycopersicum var. cerasiforme) line LC-95,
derived from an accession collected in Ecuador, harbors a natural allele (ol-2)
that confers broad-spectrum and recessively inherited resistance to powdery
mildew (Oidium neolycopersici). As both the genetic and phytopathological
characteristics of ol-2-mediated resistance are reminiscent of powdery mildew
immunity conferred by loss-of-function mlo alleles in barley and Arabidopsis, we
initiated a candidate-gene approach to clone Ol-2. A tomato Mlo gene (SlMlo1)
with high sequence-relatedness to barley Mlo and Arabidopsis AtMLO2 mapped to the
chromosomal region harboring the Ol-2 locus. Complementation experiments using
transgenic tomato lines as well as virus-induced gene silencing assays suggested
that loss of SlMlo1 function is responsible for powdery mildew resistance
conferred by ol-2. In progeny of a cross between a resistant line bearing ol-2
and the susceptible tomato cultivar Moneymaker, a 19-bp deletion disrupting the
SlMlo1 coding region cosegregated with resistance. This polymorphism results in a
frameshift and, thus, a truncated nonfunctional SlMlo1 protein. Our findings
reveal the second example of a natural mlo mutant that possibly arose
post-domestication, suggesting that natural mlo alleles might be evolutionarily
short-lived due to fitness costs related to loss of mlo function.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18052880 [PubMed - indexed for MEDLINE]

163: J Microbiol Biotechnol. 2007 Feb;17(2):335-41.

Qualitative and quantitative analysis of genetically modified pepper.

Song HS, Kim JH, Kim DH, Kim HY.

Institute of Life Science & Resources and Graduate School of Biotechnology, Kyung
Hee University, Suwon 446-701, Korea.

For the development of qualitative and quantitative PCR methods of genetically
modified (GM) pepper developed in Korea, a capsanthin-capsorubin synthase (CCS)
gene was used as the endogenous reference gene. The primer pair ccs-F/R
amplifying the pepper endogenous gene gave rise to an amplicon of 102 bp. No
amplified product was observed when DNA samples from 16 different plants were
used as templates. The construct-specific primer pairs amplifying the junction
region of the bar gene and Ti7 introduced in GM pepper gave rise to an amplicon
of 182 bp. Quantitative PCR assay was performed using a TaqMan probe and a
standard plasmid as a reference molecule, which contained both an endogenous and
event-specific sequence. For the validation of this method, the test samples
containing 0.1, 1, 3, 5, and 10% GM pepper were quantified.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18051766 [PubMed - indexed for MEDLINE]

164: J Microbiol Biotechnol. 2007 Apr;17(4):681-4.

Quantitative analysis of phosphinothricin-N-acetyltransferase in genetically
modified herbicide tolerant pepper by an enzyme-linked immunosorbent assay.

Shim YY, Shin WS, Moon GS, Kim KH.

Saskatoon Research Centre, Agriculture and Agri-Food Canada, 107 Science Place,
Saskatoon, Saskatchewan S7N OX2, Canada.

An immunoassay method was developed to quantitatively detect
phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance
(bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was
overexpressed in Escherichia coli M15 (pQE31-bar) and efficiently purified by
Ni2+ affinity chromatography. A developed sandwich enzyme-linked immunosorbent
assay (S-ELISA) method (detection limit: 0.01 microg/ml) was 100-fold more
sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot
analysis in detecting the recombinant PAT. In real sample tests, PAT in
genetically modified herbicide-tolerant (GMHT) peppers was successfully
quantified [4.9 +/- 0.4 microg/g of sample (n = 6)] by the S-ELISA method. The
S-ELISA method developed here could be applied to other GMHT crops and vegetables
producing PAT.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18051284 [PubMed - indexed for MEDLINE]

165: J Microbiol Biotechnol. 2007 Apr;17(4):547-59.

Bacillus thuringiensis as a specific, safe, and effective tool for insect pest
control.

Roh JY, Choi JY, Li MS, Jin BR, Je YH.

Department of Agricultural Biotechnology, College of Agriculture and Life
Sciences, Seoul National University, Seoul 151-742, Korea.

Bacillus thuringiensis (Bt) was first described by Berliner [10] when he isolated
a Bacillus species from the Mediterranean flour moth, Anagasta kuehniella, and
named it after the province Thuringia in Germany where the infected moth was
found. Although this was the first description under the name B. thuringiensis,
it was not the first isolation. In 1901, a Japanese biologist, Ishiwata
Shigetane, discovered a previously undescribed bacterium as the causative agent
of a disease afflicting silkworms. Bt was originally considered a risk for
silkworm rearing but it has become the heart of microbial insect control. The
earliest commercial production began in France in, 1938, under the name Sporeine
[72]. A resurgence of interest in Bt has been attributed to Edward Steinhaus
[105], who obtained a culture in 1942 and attracted attention to the potential of
Bt through his subsequent studies. In 1956, T. Angus [3] demonstrated that the
crystalline protein inclusions formed in the course of sporulation were
responsible for the insecticidal action of Bt. By the early 1980's, Gonzalez et
al. [48] revealed that the genes coding for crystal proteins were localized on
transmissible plasmids, using a plasmid curing technique, and Schnepf and
Whiteley [103] first cloned and characterized the genes coding for crystal
proteins that had toxicity to larvae of the tobacco hornworm, from plasmid DNA of
Bt subsp. kurstaki HD-1. This first cloning was followed quickly by the cloning
of many other cry genes and eventually led to the development of Bt transgenic
plants. In the 1980s, several scientists successively demonstrated that plants
can be genetically engineered, and finally, Bt cotton reached the market in 1996
[104].

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 18051264 [PubMed - indexed for MEDLINE]

166: Plant Mol Biol. 2008 Feb;66(3):289-301. Epub 2007 Nov 30.

Functional analysis of rice HOMEOBOX4 (Oshox4) gene reveals a negative function
in gibberellin responses.

Dai M, Hu Y, Ma Q, Zhao Y, Zhou DX.

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural
University, Wuhan 430070, China.

The homeodomain-leucine zipper (HD-Zip) putative transcription factor genes are
divided into 4 families. In this work, we studied the function of a rice HD-Zip I
gene, H OME O BO X4 (Oshox4). Oshox4 transcripts were detected in leaf and floral
organ primordia but excluded from the shoot apical meristem and the protein was
nuclear localized. Over-expression of Oshox4 in rice induced a semi-dwarf
phenotype that could not be complemented by applied GA3. The over-expression
plants accumulated elevated levels of bioactive GA, while the GA catabolic gene
GA2ox3 was upregulated in the transgenic plants. In addition, over-expression of
Oshox4 blocked GA-dependent alpha-amylase production. However, down-regulation of
Oshox4 in RNAi transgenic plants induced no phenotypic alteration. Interestingly,
the expression of YAB1 that is involved in the negative feedback regulation of
the GA biosynthesis was upregulated in the Oshox4 over-expressing plants.
One-hybrid assays showed that Oshox4 could interact with YAB1 promoter in yeast.
In addition, Oshox4 expression was upregulated by GA. These data together suggest
that Oshox4 may be involved in the negative regulation of GA signalling and may
play a role to fine tune GA responses in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18049796 [PubMed - indexed for MEDLINE]

167: Science. 2007 Nov 30;318(5855):1417.

Plants tolerant of high boron levels.

Miwa K, Takano J, Omori H, Seki M, Shinozaki K, Fujiwara T.

Biotechnology Research Center, University of Tokyo, Tokyo 113-8657, Japan.

Reduced crop productivity due to soils containing toxic levels of boron (B) is a
worldwide problem in food production. It is estimated that up to 17% of the
barley yield losses in southern Australia are caused by B toxicity. We found that
the expression of AtBOR4, an Arabidopsis paralog of BOR1, the first identified
boron transporter gene, generates plants that are tolerant of high B levels. BOR4
is a polarly localized borate exporter that enhances B efflux from roots. The
present study is a foundation for the improvement of crop productivity in soils
containing excess B, which are distributed in arid areas of the world.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18048682 [PubMed - indexed for MEDLINE]

168: Proc Natl Acad Sci U S A. 2007 Dec 4;104(49):19631-6. Epub 2007 Nov 28.

Delayed leaf senescence induces extreme drought tolerance in a flowering plant.

Rivero RM, Kojima M, Gepstein A, Sakakibara H, Mittler R, Gepstein S, Blumwald E.

Department of Plant Sciences, University of California, Davis, CA 95616, USA.

Drought, the most prominent threat to agricultural production worldwide,
accelerates leaf senescence, leading to a decrease in canopy size, loss in
photosynthesis and reduced yields. On the basis of the assumption that senescence
is a type of cell death program that could be inappropriately activated during
drought, we hypothesized that it may be possible to enhance drought tolerance by
delaying drought-induced leaf senescence. We generated transgenic plants
expressing an isopentenyltransferase gene driven by a stress- and
maturation-induced promoter. Remarkably, the suppression of drought-induced leaf
senescence resulted in outstanding drought tolerance as shown by, among other
responses, vigorous growth after a long drought period that killed the control
plants. The transgenic plants maintained high water contents and retained
photosynthetic activity (albeit at a reduced level) during the drought. Moreover,
the transgenic plants displayed minimal yield loss when watered with only 30% of
the amount of water used under control conditions. The production of
drought-tolerant crops able to grow under restricted water regimes without
diminution of yield would minimize drought-related losses and ensure food
production in water-limited lands.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18048328 [PubMed - indexed for MEDLINE]

169: J Biochem Mol Biol. 2007 Nov 30;40(6):952-8.

Characterization of a stress-responsive ankyrin repeat-containing zinc finger
protein of Capsicum annuum (CaKR1).

Seong ES, Choi D, Cho HS, Lim CK, Cho HJ, Wang MH.

School of Biotechnology, Kangwon National Uiversity, Chuncheon, Kangwon-do
200-701, Korea.

We isolated many genes induced from pepper cDNA microarray data following their
infection with the soybean pustule pathogen Xanthomonas axonopodis pv. glycines
8ra. A full-length cDNA clone of the Capsicum annuum ankyrin-repeat domain
C(3)H(1) zinc finger protein (CaKR1) was identified in a chili pepper using the
expressed sequence tag (EST) database. The deduced amino acid sequence of CaKR1
showed a significant sequence similarity (46%) to the ankyrin-repeat protein in
very diverse family of proteins of Arabidopsis. The gene was induced in response
to various biotic and abiotic stresses in the pepper leaves, as well as by an
incompatible pathogen, such as salicylic acid (SA) and ethephon. CaKR1 expression
was highest in the root and flower, and its expression was induced by treatment
with agents such as NaCl and methyl viologen, as well as by cold stresses. These
results showed that CaKR1 fusion with soluble, modified green fluorescent protein
(smGFP) was localized to the cytosol in Arabidopsis protoplasts, suggesting that
CaKR1 might be involved in responses to both biotic and abiotic stresses in
pepper plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18047791 [PubMed - indexed for MEDLINE]

170: J Neuroendocrinol. 2008 Feb;20(2):182-7. Epub 2007 Nov 28.

Changes in the brain serotonin satiety system in transgenic rats lacking brain
angiotensinogen.

Voigt JP, Raasch W, Hö¶rtnagl H, Bader M, Fink H, Jö¶hren O.

University of Nottingham, School of Veterinary Medicine and Science, Sutton
Bonington Campus, Sutton Bonington, Leicestershire, UK.
peter.voigt@nottingham.ac.uk

In transgenic rats, TGR(ASrAOGEN)680, with reduced glial expression of
angiotensinogen, changes in brain angiotensinogen are associated with reductions
in serotonin (5-HT) content and/or 5-HT metabolism as determined in various brain
regions, including the hypothalamus. These rats showed an anxious phenotype upon
a first behavioural screen. The present study aimed to extend the search for
functional consequences of changes in brain 5-HT with respect to feeding
behaviour in these transgenic rats. In feeding experiments, rats were treated
with the anorectic drug fenfluramine to probe for functional changes in the
serotonergic satiety system. Fenfluramine (0.3 mg/kg, i.p.) reduced food intake
in TGR(ASrAOGEN)680 rats whereas the minimal effective dose in wild-type rats was
3 mg/kg, i.p. Although, in the cortex, no differences were apparent in the
expression of serotonin 5-HT(1A), 5-HT(1B), 5-HT(2C) receptor and 5-HT
transporter mRNAs between TGR(ASrAOGEN)680 and wild-type rats, the expression of
mRNAs for the 5-HT(2C) receptor and 5-HT transporter mRNA were significantly
higher in the hypothalamus of TGR(ASrAOGEN)680 rats compared to wild-type rats.
No differences were found in the mRNA levels for hypothalamic 5-HT(1A) and
5-HT(1B) receptors between TGR(ASrAOGEN)680 and wild-type rats. Taken together,
these findings suggest that the transgenic effect on the brain 5-HT system is
paralleled by functional changes of the serotonergic feeding system.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18047554 [PubMed - indexed for MEDLINE]

171: J Appl Microbiol. 2007 Dec;103(6):2593-600.

Immigration of Bacillus thuringiensis to bean leaves from soil inoculum or distal
plant parts.

Maduell P, Armengol G, Llagostera M, Lindow S, Orduz S.

Biotechnology and Biological Control Unit, Corporaciö¨n para Investigaciones
Biolö¨gicas, Medellö­n, Colombia.

AIMS: We addressed the process of immigration of Bacillus thuringiensis from soil
to leaves and its capacity to grow on bean diffusate medium (BDM), a medium
designed to simulate the nutrient composition of the phylloplane. METHODS AND
RESULTS: Two different B. thuringiensis strains were inoculated into soils, onto
seeds or onto lower leaves of bean plants to determine if they were able to
disperse to upper leaves under controlled conditions. While B. thuringiensis
isolates were commonly recovered from leaves exposed to such inocula, populations
were very low (<10 CFU cm(-2) of leaf). In addition, the number of cells of B.
thuringiensis recovered decreased with increasing distance from the soil or from
the inoculated leaves. Moreover, B. thuringiensis colonies did not grow well on
BDM. CONCLUSIONS: This indicates that B. thuringiensis disperses poorly from the
soil or the seed to the leaves or between leaves of the same plant under
controlled conditions. Bacillus thuringiensis apparently has greater nutrient
requirements than other bacterial species that are prominent inhabitants of the
phylloplane. SIGNIFICANCE AND IMPACT OF THE STUDY: Finding the mechanisms that
favour bacteria colonization on leaves will in turn help to improve the efficacy
of biocontrol agents against the target pests.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18045443 [PubMed - indexed for MEDLINE]

172: J Appl Microbiol. 2007 Dec;103(6):2248-57.

The heterologous expression of polysaccharidase-encoding genes with oenological
relevance in Saccharomyces cerevisiae.

van Rensburg P, Strauss ML, Lambrechts MG, Cordero Otero RR, Pretorius IS.

Department of Viticulture and Oenology, Institute for Wine Biotechnology,
Stellenbosch University, Matieland, South Africa.

AIMS: The main objective of this study was to develop polysaccharide-degrading
wine strains of Saccharomyces cerevisiae, which are able to improve aspects of
wine processing and clarification, as well as colour extraction and stabilization
during winemaking. METHODS AND RESULTS: Two yeast expression/secretion gene
cassettes were constructed, namely (i) a pectinase gene cassette (pPPK)
consisting of the endo-polygalacturonase gene (pelE) from Erwinia chrysanthemi
and the pectate lyase gene (peh1) from Erwinia carotovora and (ii) a
glucanase/xylanase gene cassette (pEXS) containing the endo-beta-1,4-glucanase
gene (end1) from Butyrivibrio fibrisolvens and the endo-beta-1,4-xylanase gene
(xynC) from Aspergillus niger. The commercial wine yeast strain, VIN13, was
transformed separately with these two gene cassettes and checked for the
production of pectinase, glucanase and xylanase activities. Pinot Noir, Cinsaut
and Muscat d'Alexandria grape juices were fermented using the VIN13[pPPK]
pectinase- and the VIN13[pEXS] glucanase/xylanase-producing transformants.
Chemical analyses of the resultant wines indicated that (i) the
pectinase-producing strain caused a decrease in the concentration of phenolic
compounds in Pinot Noir whereas the glucanase/xylanase-producing strain caused an
increase in phenolic compounds presumably because of the degradation of the grape
skins; (ii) the glucanase/xylanase-producing strain caused a decrease in wine
turbidity, especially in Pinot Noir wine, as well as a clear increase in colour
intensity and (iii) in the Muscat d'Alexandria and Cinsaut wines, the differences
between the control wines (fermented with the untransformed VIN3 strain) and the
wines produced by the two transformed strains were less prominent showing that
the effect of these polysaccharide-degrading enzymes is cultivar-dependent.
CONCLUSIONS: The recombinant wine yeasts producing pectinase, glucanase and
xylanase activities during the fermentation of Pinot Noir, Cinsaut and Muscat
d'Alexandria grape juice altered the chemical composition of the resultant wines
in a way that such yeasts could potentially be used to improve the clarity,
colour intensity and stability and aroma of wine. SIGNIFICANCE AND IMPACT OF THE
STUDY: Aspects of commercial-scale wine processing and clarification, colour
extraction and stabilization, and aroma enhancement could potentially be improved
by the use of polysaccharide-degrading wine yeasts without the addition of
expensive commercial enzyme preparations. This offers the potential to further
improve the price:quality ratio of wine according to consumer expectations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18045408 [PubMed - indexed for MEDLINE]

173: J Agric Food Chem. 2007 Dec 26;55(26):10827-31. Epub 2007 Nov 29.

Strong increase of foliar inulin occurs in transgenic lettuce plants (Lactuca
sativa L.) overexpressing the Asparagine Synthetase A gene from Escherichia coli.

Sobolev AP, Segre AL, Giannino D, Mariotti D, Nicolodi C, Brosio E, Amato ME.

Institute of Chemical Methodologies, CNR, Research Area of Rome, Italy.
anatoli.sobolev@imc.cnr.it

Transgenic lettuce (Lactuca sativa L. cv. 'Cortina') lines expressing the
asparagine synthetase A gene from Escherichia coli were produced to alter the
plant nitrogen status and eventually enhance growth. The relative molecular
abundance of water-soluble metabolites was measured by 1H NMR in transgenic and
conventional plants at early developmental stages and grown under the same
conditions. NMR metabolic profiles assessed that a transgenic line and the
wild-type counterpart shared the same compounds, but it also revealed side
effects on the carbon metabolism following genetic modification. Concerning the
nitrogen status, the amino acid content did not vary significantly, except for
glutamic acid and gamma-aminobutyric acid, which diminished in the transgenics.
As for the carbon metabolism, in transgenic leaves the contents of sucrose,
glucose, and fructose decreased, whereas that of inulin increased up to 30 times,
accompanied by the alteration of most Krebs's cycle organic acids and the rise of
tartaric acid compared to nontransformed controls. Lettuce leaf inulins consisted
of short oligomeric chains made of one glucose unit bound to two/four fructose
units. Inulins are beneficial for human health, and they are extracted from
plants and commercialized as long-chain types, whereas the short forms are
synthesized chemically. Hence, lettuce genotypes with high content of foliar
short-chain inulin represent useful materials for breeding strategies and a
potential source for low molecular weight inulin.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18044837 [PubMed - indexed for MEDLINE]

174: J Appl Toxicol. 2008 Mar;28(2):217-26.

Pancreatic response of rats fed genetically modified soybean.

Magaö±a-Gö¨mez JA, Cervantes GL, Yepiz-Plascencia G, de la Barca AM.

Coordinaciö¨n de Nutriciö¨n, Centro de Investigaciö¨n en Alimentaciö¨n y Desarrollo,
A.C. P.O. Box 1735, Sonora 83000, Mexico.

Mice fed genetically modified (GM) soybean were not affected in nutritional
performance, but pancreatic microscopic features were disturbed. The mechanisms
for these contradictory findings are unknown. This study analysed the histology
of acinar pancreatic cells and the expression of pancreatitis-associated protein
(PAP) and trypsinogen mRNA in rats fed GM soy protein. Two bioassays were run,
each one with 34 Wistar rats distributed into two groups fed with non-GM or
GM-soy protein (18% protein) for 0, 1, 3, 5, 15 and 30 days. Nutritional
evaluation, plasma amylase levels, pancreatic histological analysis and
quantification of PAP and trypsinogen mRNAs levels using quantitative real-time
RT-PCR were done. No differences in nutritional performance among rats fed non-GM
and GM diets were found. The GM, but not the non-GM, diet induced zymogen-granule
depletion after 15 days feeding, returning to normal levels after 30 days (P <
0.05). Acinar disorganization started as early as 5 days after initiation of the
GM diet and it recovered after 30 days. Levels of PAP mRNA significantly
increased in the GM diet between day 1 and day 3 and decreased to the basal level
by day 15. Trypsinogen mRNA peaked at two different times; at day 1 and at day
15, decreasing to basal levels after 30 days. Plasma amylase levels remained
unchanged at all times. This indicates that GM soy protein intake affected
pancreas function, evidenced by the early acute PAP mRNA increased levels and
pancreas cellular changes followed by recuperation of acinar cells after 30 days.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18041736 [PubMed - indexed for MEDLINE]

175: Plant J. 2008 Mar;53(6):895-908. Epub 2007 Nov 23.

Heterologous expression, and biochemical and cellular characterization of CaPLA1
encoding a hot pepper phospholipase A1 homolog.

Seo YS, Kim EY, Mang HG, Kim WT.

Department of Biology, College of Science, Yonsei University, Seoul 120-749,
Korea.

Phospholipid signaling has been recently implicated in diverse cellular processes
in higher plants. We identified a cDNA encoding the phospholipase A1 homolog
(CaPLA1) from 5-day-old early roots of hot pepper. The deduced amino acid
sequence showed that the lipase-specific catalytic triad is well conserved in
CaPLA1. In vitro lipase assays and site-directed mutagenesis revealed that CaPLA1
possesses PLA1 activity, which catalyzes the hydrolysis of phospholipids at the
sn-1 position. CaPLA1 was selectively expressed in young roots, at days 4-5 after
germination, and rapidly declined thereafter, suggesting that the expression of
CaPLA1 is subject to control by a development-specific mechanism in roots.
Because transgenic work was extremely difficult in hot peppers, in this study we
overexpressed CaPLA1 in Arabidopsis so as to provide cellular information on the
function of this gene. CaPLA1 overexpressors had significantly longer roots,
leaves and petioles, and grew more rapidly than the wild-type plants, leading to
an early bolting phenotype with prolonged inflorescence. Microscopic analysis
showed that the vegetative tissues of 35S:CaPLA1 plants contained an increased
number of small-sized cells, which resulted in highly populated cell layers. In
addition, mRNAs for cell cycle-controlled proteins and fatty acid catabolizing
enzymes were coordinately upregulated in CaPLA1-overexpressing plants. These
results suggest that CaPLA1 is functionally relevant in heterologous Arabidopsis
cells, and hence might participate in a subset of positive control mechanisms of
cell and tissue growth in transgenic lines. We discuss possible biochemical and
cellular functions of CaPLA1 in relation to the phospholipid signaling pathway in
hot pepper and transgenic Arabidopsis plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18036200 [PubMed - indexed for MEDLINE]

176: Food Chem Toxicol. 2008 Apr;46(4):1213-20. Epub 2007 Oct 18.

Liver and colon DNA oxidative damage and gene expression profiles of rats fed
Arabidopsis thaliana mutant seeds containing contrasted flavonoids.

Luceri C, Giovannelli L, Pitozzi V, Toti S, Castagnini C, Routaboul JM, Lepiniec
L, Larrosa M, Dolara P.

Department of Preclinical and Clinical Pharmacology, University of Florence,
Viale Pieraccini 6, 50139 Florence, Italy. cristina.luceri@unifi.it


Plant polyphenols, such as flavonoids, comprise many compounds, ranging from
simple phenolic molecules (i.e. flavonols, anthocyanins) to polymeric structures
with high molecular weight (as proanthocyanidins, PAs). We investigated the
effects of flavonoids by feeding Wistar rats Arabidopsis thaliana seeds carrying
mutations in key enzymes of the flavonoid biosynthetic pathway (15% w/w seeds for
4 weeks). The seeds used were: Ws-2 wild-type containing flavonols and PAs, tt3-4
mutant containing flavonols only, ban-5 accumulating flavonols and anthocyanins,
tt4-8 mutant, deprived of flavonoids. DNA oxidative damage was significantly
reduced only in the liver of rats fed tt3-4 mutant seeds. Microarray analysis of
the liver revealed down-regulation of genes associated with oxidative stress,
Krebs cycle, electron transport and proteasome degradation in all experimental
groups compared to the tt4-8-fed reference rats; therefore, these effects were
due to the flavonol content and not to high molecular weight compounds. We
observed a down-regulation of inflammatory response genes in the colon mucosa in
ban-5- fed rats, probably due to anthocyanin content. In conclusion, flavonols
exhibited antioxidant effects at systemic level, whereas high molecular weight
flavonoids affected only the colon, probably due to their limited absorption.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18035473 [PubMed - indexed for MEDLINE]

177: FEMS Microbiol Ecol. 2008 Jan;63(1):118-31. Epub 2007 Nov 20.

Prevalence of pSmeSM11a-like plasmids in indigenous Sinorhizobium meliloti
strains isolated in the course of a field release experiment with genetically
modified S. meliloti strains.

Kuhn S, Stiens M, Pö¼hler A, Schlö¼ter A.

Lehrstuhl fö¼r Genetik, Fakultö¤t fö¼r Biologie, Universitö¤t Bielefeld, Bielefeld,
Germany.

Plasmid pSmeSM11a, residing in the indigenous Sinorhizobium meliloti strain SM11
originating from a field in Strassmoos (Bavaria, Germany), was analysed
previously at the genomic level. Thirty-seven indigenous S. meliloti strains,
originating from two different locations in Germany, were screened for genes
identified previously on pSmeSM11a. Seven of these strains harbour accessory
plasmids that are very similar to pSmeSM11a. The identified pSmeSM11a-like
plasmids are c. 130-150 kb in size and possess nearly identical restriction
profiles. Up to 30 genes identified previously on pSmeSM11a could be detected on
these plasmids by hybridisation experiments, e.g., the nodulation genes nodP and
nodQ, the ethylene level modulation gene acdS and the taurine metabolism gene
tauD. A few pSmeSM11a genes were also detected on other plasmids. The reference
plasmid pSmeSM11a contains a region that is similar to a segment of S. meliloti
strain Rm1021 pSymA. Regions with similarity to pSymA were also detected on the
aforementioned seven pSmeSM11a-like plasmids. The specifications of these regions
are nearly identical to the one on pSmeSM11a and differ from Rm1021 pSymA as
determined by nucleotide sequence analysis. Two further plasmids similar to
pSmeSM11a completely lack the pSymA-region. Those strains carrying accessory
plasmids that contain the acdS gene encoding 1-aminocyclopropane-1-carboxylate
deaminase are able to grow on 1-aminocyclopropane-1-carboxylate as the sole
source of nitrogen, demonstrating functionality of the acdS gene product. About
36% of the analysed plasmids, including three pSmeSM11a-like plasmids, could be
transferred to another S. meliloti recipient strain, allowing for their
dissemination in S. meliloti populations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18034835 [PubMed - indexed for MEDLINE]

178: J Food Sci. 2007 Nov;72(9):S689-95.

Zinc and iron bioavailability of genetically modified soybeans in rats.

Martino HS, Martin BR, Weaver CM, Bressan J, Esteves EA, Costa NM.

Dept. de Nutriö§ö¸o e Saöºde, Univ. Federal de Viö§osa, Viö§osa-MG 36.570-000, Brazil.

The aim of this work was to evaluate zinc and iron bioavailability of UFV-116, a
new variety without 2 lipoxygenases, with better taste and flavor than a
commercial variety OCEPAR 19, containing all 3 isozymes. To evaluate zinc
absorption using 65Zn whole body retention and femur 65Zn uptake, rats were given
3 g of a 65ZnCl2 labeled test meal (0.25 microCi). The 2 varieties were tested at
the level of 9 and 30 ppm of zinc as defatted soy flour. Two other groups
(control) received egg white as source of protein and ZnS04.H20 as the zinc
source. To evaluate iron absorption, using 59Fe whole body retention, animals
were given a 3 g 59FeCl3 labeled test meal (0.2 microCi). The 2 varieties were
tested at 12 and 25 ppm iron as defatted soy flour. Whole fat soy flour of
variety 1 (UFV-116) was higher (P < 0.05) in Ca, K, Mg, phytic acid, and oxalate
than variety 2 (OCEPAR-19). No difference was observed among the soybean
varieties (P > 0.05) for femur 65Zn retention, at different levels of zinc.
However, whole body retention was lower (P < 0.05) for UFV-116 than for
OCEPAR-19. Femur 65Zn uptake was correlated with the whole body retention;
however, whole body retention was more sensitive. Whole body 59Fe retention from
UFV-116 was lower (P < 0.05) than from OCEPAR-19. Zinc and iron bioavailability
was lower for UFV-116, possibly due to its higher content of antinutrient
factors, especially phytate.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18034754 [PubMed - indexed for MEDLINE]

179: J Food Sci. 2007 Nov;72(9):R131-7.

Nutritional and safety assessments of foods and feeds nutritionally improved
through biotechnology: case studies: executive summary of a task force report by
the International Life Sciences Institute, Washington, D.C.

International Life Sciences Institute.

During the last 2 decades, the public and private sectors have made substantial
international research progress toward improving the nutritional value of a wide
range of food and feed crops. Nevertheless, significant numbers of people still
suffer from the effects of undernutrition. In addition, the nutritional quality
of feed is often a limiting factor in livestock production systems, particularly
those in developing countries. As newly developed crops with nutritionally
improved traits come closer to being available to producers and consumers, we
must ensure that scientifically sound and efficient processes are used to assess
the safety and nutritional quality of these crops. Such processes will facilitate
deploying these crops to those world areas with large numbers of people who need
them. This document describes 5 case studies of crops with improved nutritional
value. These case studies examine the principles and recommendations published by
the Intl. Life Sciences Inst. (ILSI) in 2004 for the safety and nutritional
assessment of foods and feeds derived from nutritionally improved crops (ILSI
2004). One overarching conclusion that spans all 5 case studies is that the
comparative safety assessment process is a valid approach. Such a process has
been endorsed by many publications and organizations, including the 2004 ILSI
publication. The type and extent of data that are appropriate for a
scientifically sound comparative safety assessment are presented on a
case-by-case basis in a manner that takes into account scientific results
published since the 2004 ILSI report. This report will appear in the January
issue of Comprehensive Reviews in Food Science and Food Safety.

Publication Types:
Guideline

PMID: 18034742 [PubMed - indexed for MEDLINE]

180: Toxicol Sci. 2008 Mar;102(1):100-9. Epub 2007 Nov 21.

Differences in allergenic potential of food extracts following oral exposure in
mice reflect differences in digestibility: potential approaches to safety
assessment.

Bowman CC, Selgrade MK.

Immunotoxicology Branch, Experimental Toxicology Division, National Health and
Environmental Effects Research Laboratory, U.S. Environmental Protection Agency,
Research Triangle Park, North Carolina 27711, USA. bowman.christal@epa.gov

An animal model for food allergy is needed to assess genetically modified food
crops for potential allergenicity. The ideal model must produce allergic antibody
(IgE) to proteins differentially according to known allergenicity before being
used to accurately identify potential allergens among novel proteins. The oral
route is the most relevant for exposure to food antigens, and a protein's
stability to digestion is a current risk assessment tool based on this natural
route. However, normal laboratory animals do not mount allergic responses to
proteins administered orally due to oral tolerance, an immunologic mechanism
which specifically suppresses IgE. To circumvent oral tolerance and evoke
differential IgE responses to a panel of allergenic and nonallergenic food
extracts, female C3H/HeJ mice were exposed subcutaneously or orally with cholera
toxin as an adjuvant. All foods elicited IgE by the subcutaneous route. Oral
exposure, however, resulted in IgE to allergens (peanut, Brazil nut, and egg
white) but not to nonallergens (spinach and turkey), provided that the dose and
exposures were limited. Additionally, in vitro digestibility assays demonstrated
the presence of digestion-stable proteins in the allergenic food extracts but not
in the nonallergenic foods. Our results suggest that the subcutaneous route is
inadequate to distinguish allergens from nonallergens, but oral exposure under
the appropriate experimental conditions will result in differential allergic
responses in accordance with known allergenicity. Moreover, those foods
containing digestion-resistant proteins provoke allergic responses in this model,
supporting the current use of pepsin resistance in the decision tree for
potential allergenicity assessment.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18033772 [PubMed - indexed for MEDLINE]

181: Mol Nutr Food Res. 2007 Dec;51(12):1518-26.

Enhancing lignan biosynthesis by over-expressing pinoresinol lariciresinol
reductase in transgenic wheat.

Ayella AK, Trick HN, Wang W.

Department of Human Nutrition, Kansas State University, Manhattan, KS 66506, USA.

Lignans are phenylpropane dimers that are biosynthesized via the phenylpropanoid
pathway, in which pinoresinol lariciresinol reductase (PLR) catalyzes the last
steps of lignan production. Our previous studies demonstrated that the contents
of lignans in various wheat cultivars were significantly associated with
anti-tumor activities in APC(Min) mice. To enhance lignan biosynthesis, this
study was conducted to transform wheat cultivars ('Bobwhite', 'Madison', and
'Fielder', respectively) with the Forsythia intermedia PLR gene under the
regulatory control of maize ubiquitin promoter. Of 24 putative transgenic wheat
lines, we successfully obtained 3 transformants with the inserted ubiquitin-PLR
gene as screened by PCR. Southern blot analysis further demonstrated that
different copies of the PLR gene up to 5 were carried out in their genomes.
Furthermore, a real-time PCR indicated approximately 17% increase of PLR gene
expression over the control in 2 of the 3 positive transformants at T(0)
generation. The levels of secoisolariciresinol diglucoside, a prominent lignan in
wheat as determined by HPLC-MS, were found to be 2.2-times higher in one of the
three positive transgenic sub-lines at T(2 )than that in the wild-type (117.9 +/-
4.5 vs. 52.9 +/- 19.8 mug/g, p <0.005). To the best of our knowledge, this is the
first study that elevated lignan levels in a transgenic wheat line has been
successfully achieved through genetic engineering of over-expressed PLR gene.
Although future studies are needed for a stably expression and more efficient
transformants, the new wheat line with significantly higher SDG contents obtained
from this study may have potential application in providing additive health
benefits for cancer prevention.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 18030664 [PubMed - indexed for MEDLINE]

182: Poult Sci. 2007 Dec;86(12):2608-14.

Comparison of broiler performance and carcass parameters when fed diets
containing soybean meal produced from glyphosate-tolerant (MON 89788), control,
or conventional reference soybeans.

Taylor M, Hartnell G, Lucas D, Davis S, Nemeth M.

Monsanto Company, Product Safety Center, 800 N. Lindbergh Blvd., Mail Stop O3D,
Creve Couer, MO 63167, USA. mary.l.taylor@monsanto.com

A 42-d floor pen study was conducted to compare broiler (Ross x Ross 308)
performance and carcass measurements when fed diets containing meal produced from
glyphosate-tolerant soybeans (MON 89788) with those of broilers fed diets
containing meal produced from control soybean (A3244) that has similar genetic
background to MON 89788. Soybean meal produced from 6 conventional soybean
varieties was included in the study to provide comparison measurements for
broilers fed meal derived from conventional soybeans. It has been found that MON
89788 produces the 5-enolpyruvylshikimate-3-phosphate synthase protein from
Agrobacterium sp. strain CP4 (cp4 epsps), which confers tolerance to glyphosate,
the active ingredient in Roundup agricultural herbicides. Broilers were fed
starter diets (approximately 33% wt/wt dehulled soybean meal) from d 0 to 21 and
grower-finisher diets (approximately 30% wt/wt dehulled soybean meal) from d 21
to 42. The study utilized a randomized complete block design with 8 dietary
treatments assigned randomly within 5 blocks of 16 pens each (8 male and 8
female) with 10 birds per pen. There were 10 pens per treatment group (5 male and
5 female). No treatment differences (P > 0.05) were detected among dietary
treatments for feed intake, weight gain, adjusted feed conversion, or any
measured carcass and meat quality parameters. Comparison of all performance,
carcass, and meat quality parameters measured showed no differences (P > 0.05)
between birds fed the MON 89788 soybean meal diet and the population of birds fed
the control and 6 conventional reference soybean meal diets. It is concluded that
the diets containing soybean meal produced from MON 89788 were nutritionally
equivalent to diets containing soybean meal produced from the control and
conventional reference soybean varieties when fed to broilers.

Publication Types:
Randomized Controlled Trial

PMID: 18029807 [PubMed - indexed for MEDLINE]

183: Poult Sci. 2007 Dec;86(12):2569-81.

Comparison of broiler performance when fed diets containing event DP-356ö˜43-5
(Optimum GAT), nontransgenic near-isoline control, or commercial reference
soybean meal, hulls, and oil.

McNaughton J, Roberts M, Smith B, Rice D, Hinds M, Schmidt J, Locke M, Brink K,
Bryant A, Rood T, Layton R, Lamb I, Delaney B.

Solution BioSciences, 2028 Northwood Drive, Salisbury, MD 21801, USA.
mcnaughton@ahpharma.com

Event DP-356ö˜43-5 (356043; Optimum GAT) is a genetically modified soybean
(Glycine max) that was produced by insertion of the gat4601 and gm-hra genes. The
expression products of these genes are the glyphosate acetyltransferase 4601 and
acetolactate synthase proteins, respectively. Expression of the glyphosate
acetyltransferase 4601 protein confers tolerance in planta to the herbicidal
active ingredient glyphosate, whereas expression of the acetolactate synthase
protein confers tolerance to sulfonylurea and imidazolinone herbicides. The
objective of this study was to compare the nutritional equivalence of 356043
soybeans to nontransgenic soybeans in a 42-d feeding trial in broiler chickens.
Diets were prepared using processed fractions (meal, hulls, and oil) from
untreated 356043 soybean plants or from soybean plants treated with a mixture of
glyphosate, chlorimuron, and thifensulfuron (356043 + Gly/SU). For comparison,
additional diets were produced with soybean fractions obtained from a
nontransgenic near-isoline (control; 091) and nontransgenic commercial Pioneer
varieties (93B86, 93B15, and 93M40). Diets were fed to Ross x Cobb broilers (n =
120/group, 50% male and 50% female) in 3 phases. Starter diets contained 30%
soybean meal, grower diets 26% soybean meal, and finisher diets 21.5% soybean
meal. Soybean hulls and oil were added at 1.0 and 0.5%, respectively, across all
diets in each phase. No statistically significant differences were observed in
mortality, growth performance variables, or carcass and organ yields between
broilers consuming diets produced with 356043 or 356043 + Gly/SU soybean
fractions and those consuming diets produced with near-isoline control soybean
fractions. Additionally, all performance and carcass variables from control,
356043, and 356043 + Gly/SU soybean treatment groups fell within the tolerance
intervals constructed using data from reference soybean groups. Based on the
results from this study, it was concluded that 356043 soybean was nutritionally
equivalent to nontransgenic control soybean with a comparable genetic background.

Publication Types:
Controlled Clinical Trial

PMID: 18029803 [PubMed - indexed for MEDLINE]

184: Plant Biotechnol J. 2008 Apr;6(3):213-25. Epub 2007 Nov 19.

Regulating innovative crop technologies in Canada: the case of regulating
genetically modified crops.

Smyth S, McHughen A.

College of Biotechnology, University of Saskatchewan, 51 Campus Drive, Saskatoon,
SK, Canada S7N 5A8. stuart.smyth@usask.ca

The advent of genetically modified crops in the late 1980s triggered a regulatory
response to the relatively new field of plant genetic engineering. Over a 7-year
period, a new regulatory framework was created, based on scientific principles
that focused on risk mitigation. The process was transparent and deliberately
sought the input of those involved in crop development from non-governmental
organizations, industry, academia and federal research laboratories. The
resulting regulations have now been in place for over a decade, and the
resilience of the risk-mitigating regulations is evident as there has been no
documented case of damage to either environment or human health.

Publication Types:
Review

PMID: 18028290 [PubMed - indexed for MEDLINE]

185: Proc Natl Acad Sci U S A. 2007 Nov 27;104(48):19150-5. Epub 2007 Nov 19.

The transcription factor IDEF1 regulates the response to and tolerance of iron
deficiency in plants.

Kobayashi T, Ogo Y, Itai RN, Nakanishi H, Takahashi M, Mori S, Nishizawa NK.

Laboratory of Plant Biotechnology, Graduate School of Agricultural and Life
Sciences, University of Tokyo, 1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

Iron is essential for most living organisms and is often the major limiting
nutrient for normal growth. Plants induce iron utilization systems under
conditions of low iron availability, but the molecular mechanisms of gene
regulation under iron deficiency remain largely unknown. We identified the rice
transcription factor IDEF1, which specifically binds the iron
deficiency-responsive cis-acting element IDE1. IDEF1 belongs to an
uncharacterized branch of the plant-specific transcription factor family ABI3/VP1
and exhibits the sequence recognition property of efficiently binding to the
CATGC sequence within IDE1. IDEF1 transcripts are constitutively present in rice
roots and leaves. Transgenic tobacco plants expressing IDEF1 under the control of
the constitutive cauliflower mosaic virus 35S promoter transactivate
IDE1-mediated expression only in iron-deficient roots. Transgenic rice plants
expressing an introduced IDEF1 exhibit substantial tolerance to iron deficiency
in both hydroponic culture and calcareous soil. IDEF1 overexpression leads to the
enhanced expression of the iron deficiency-induced transcription factor gene
OsIRO2, suggesting the presence of a sequential gene regulatory network. These
findings reveal cis element/trans factor interactions that are functionally
linked to the iron deficiency response. Manipulation of IDEF1 also provides
another approach for producing crops tolerant of iron deficiency to enhance food
and biomass production in calcareous soils.

PMID: 18025467 [PubMed - indexed for MEDLINE]

186: J Dairy Sci. 2007 Dec;90(12):5706-13.

Performance of lactating dairy cows fed silage and grain from a maize hybrid with
the cry1F trait versus its nonbiotech counterpart.

Faust M, Smith B, Rice D, Owens F, Hinds M, Dana G, Hunst P.

Department of Animal Science, 123 Kildee Hall, Iowa State University, Ames 50010,
USA.

Effects of feeding grain and maize silage from a non-Bt maize and a variety of Bt
maize that contains cry1F (event TC1507, event DAS-ö˜15ö˜7-1), a gene that provides
maize with insect resistance, on the health and performance of dairy cows were
evaluated. In a crossover trial, 20 lactating Holstein cows were assigned to each
of 2 dietary treatment groups and fed diets containing whole-plant maize silage
plus maize grain from TC1507 or its near-isoline counterpart (control). Each
period of the crossover trial lasted 28 d and was preceded by a 7-d adjustment
period. To minimize variability due to stage of lactation, 2 blocks of 10 cows at
90 to 130 d of lactation at the start of the trial were used. Within each dietary
treatment, 10 cows were from each of 2 genetic selection lines (high and average
fat plus protein predicted transmitting ability). Diets were formulated to be
isocaloric and isonitrogenous. Dry matter intake and daily production of milk,
fat, protein, lactose, nonfat solids, and total solids did not differ between
cows fed the TC1507 diet and cows fed the control diet. Furthermore, milk from
cows in different dietary treatment groups did not differ in milk urea nitrogen
concentration or somatic cell count. For milk fat percentage, a significant
dietary treatment by genetic group interaction was detected although overall
yield of milk and solids-corrected milk did not differ with diet. Physical
measures of cow health including body weight, body condition score, temperature,
pulse, and respiration rate were collected weekly; dietary treatment group means
for these measures were not different. Blood chemistry and hematological analyses
were conducted using blood samples collected from cows before the start of the
trial and at the end of each period. Overall, the TC1507 and control groups did
not differ in any of these indices of health status. Further, hematological
profiles for cows in the dietary treatment groups were not different. In summary,
no differences were detected in milk production, milk composition, or cow health
as indicated by physical measures, blood chemistry, and hematological analyses
between dairy cows fed diets containing maize grain plus whole-plant maize silage
from TC1507 and dairy cows fed grain plus silage from its near-isoline
counterpart.

Publication Types:
Randomized Controlled Trial

PMID: 18024763 [PubMed - indexed for MEDLINE]

187: Toxicol Lett. 2007 Dec 10;175(1-3):118-35. Epub 2007 Oct 10.

Zero tolerances in food and animal feed -- are there any scientific alternatives?
A European point of view on an international controversy.

Heberer T, Lahrssen-Wiederholt M, Schafft H, Abraham K, Pzyrembel H, Henning KJ,
Schauzu M, Braeunig J, Goetz M, Niemann L, Gundert-Remy U, Luch A, Appel B,
Banasiak U, Bö¶l GF, Lampen A, Wittkowski R, Hensel A.

Federal Institute for Risk Assessment, Section 55, - Residues of Medicinal
Products, Diedersdorfer Weg 1, 12277 Berlin, Germany. bfr@bfr.bund.de

A number of zero tolerance provisions are contained in both food and animal feed
law, e.g. for chemical substances whose occurrence is not permitted or is
directly prohibited in food or animal feed. In the European Union, bans of this
kind were introduced to give consumers and animals the greatest possible
protection from substances with a possible hazard potential within the intendment
of the hazard prevention principles and current precautionary measures. This also
applies to substances for which an acceptable daily intake cannot be derived and
a maximum residue limit cannot, therefore, be established, e.g. due to missing or
inadequate toxicological data. Zero tolerances are also under discussion as trade
barriers because their use has triggered numerous legal disputes. This paper
draws together the results of an evaluation of alternative risk assessment
methods to be used for the risk assessment of substances to which currently only
zero tolerances apply. It will demonstrate that, depending on the available
toxicological data, a scientifically sound risk assessment may still be possible.
In this context, the two concepts - margin of exposure and threshold of
toxicological concern - are very promising approaches. Until the scientific and
sociopolitical discussions have been completed, it is essential that the
principle of zero tolerances be upheld, especially for those substances which may
be genotoxic carcinogens. In microbiology, there is no legal room for manoeuvre
with regard to food safety criteria established for reasons of consumer health
protection on the basis of scientific assessments.

PMID: 18024010 [PubMed - indexed for MEDLINE]

188: Toxicol Lett. 2007 Dec 10;175(1-3):82-8. Epub 2007 Oct 7.

A rapid and inexpensive method to screen for common foods that reduce the action
of acrylamide, a harmful substance in food.

Hasegawa K, Miwa S, Tajima T, Tsutsumiuchi K, Taniguchi H, Miwa J.

Institute for Biological Function, Chubu University, 1200 Matsumoto, Kasugai
487-8501, Japan.

By DNA microarray and protein 2-DE screens for Caenorhabditis elegans genes
up-regulated by acrylamide, we selected the gst-4 gene and constructed a gst::gfp
fusion gene, which was used to transform C. elegans into a biosensor for
acrylamide. This biosensor detects acrylamide as a GFP-expression signal in a
dose- and time-dependent manner. When the biosensor was exposed to acrylamide
together with commercially available powdered green tea, GFP levels decreased to
the control level, suggestive of acrylamide detoxification or prevention of GST
induction. The present methodology should be applicable for screening of not only
harmful substances but also substances that reduce or counteract their
harmfulness or action, with appropriately constructed visible biosensors.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18023302 [PubMed - indexed for MEDLINE]

189: J Agric Food Chem. 2007 Dec 26;55(26):10714-9. Epub 2007 Nov 20.

Effect of a Bowman-Birk proteinase inhibitor from Phaseolus coccineus on
Hypothenemus hampei gut proteinases in vitro.

de Azevedo Pereira R, Valencia-Jimö©nez A, Magalhö¸es CP, Prates MV, Melo JA, de
Lima LM, de Sales MP, Tempel Nakasu EY, da Silva MC, Grossi-de-Sö¡ MF.

Embrapa Recursos Genö©ticos e Biotecnologia, PqEB, W5 Norte Final, Brasö­lia - DF,
70770-900, Brazil.

The coffee berry borer, Hypothenemus hampei (Ferrari), is an important
devastating coffee pest worldwide. Both trypsin and chymotrypsin enzyme
activities from H. hampei larval midgut can be inactivated by proteinaceous
enzyme-inhibitors. A serine proteinase inhibitor belonging to the Bowman-Birk
class was purified from a wild accession of Phaseolus coccineus L. seeds. The
inhibitor (PcBBI1) is a cysteine-rich protein that is heat-stable at alkaline pH.
MALDI-TOF/MS analysis showed that PcBBI1 occurs in seeds as a monomer (8689 Da)
or dimer (17,378 Da). Using in vitro inhibition assays, it was found that PcBBI1
has a high inhibitory activity against H. hampei trypsin-like enzymes, bovine
pancreatic chymotrypsin, and trypsin. According to this, PcBBI1 could be a
promising tool to make genetically modified coffee with resistance to coffee
berry borer.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18020416 [PubMed - indexed for MEDLINE]

190: Commun Agric Appl Biol Sci. 2007;72(1):177-81.

Modelling heterogeneity to estimate the ex ante value of biotechnology
innovations.

Dillen K, Demont M, Tollens E.

Centre for Agricultural and food economics Dept. of land management and
economics, K.U. Leuven.

PMID: 18018883 [PubMed - indexed for MEDLINE]

191: Genes Dev. 2007 Nov 15;21(22):2874-9.

Riboswitch-dependent gene regulation and its evolution in the plant kingdom.

Bocobza S, Adato A, Mandel T, Shapira M, Nudler E, Aharoni A.

Department of Plant Sciences, Weizmann Institute of Science, Rehovot 76100,
Israel.

Riboswitches are natural RNA sensors that affect gene control via their capacity
to bind small molecules. Their prevalence in higher eukaryotes is unclear. We
discovered a post-transcriptional mechanism in plants that uses a riboswitch to
control a metabolic feedback loop through differential processing of the
precursor RNA 3' terminus. When cellular thiamin pyrophosphate (TPP) levels rise,
metabolite sensing by the riboswitch located in TPP biosynthesis genes directs
formation of an unstable splicing product, and consequently TPP levels drop. When
transformed in plants, engineered TPP riboswitches can act autonomously to
modulate gene expression. In an evolutionary perspective, a TPP riboswitch is
also present in ancient plant taxa, suggesting that this mechanism is active
since vascular plants emerged 400 million years ago.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18006684 [PubMed - indexed for MEDLINE]

192: Trends Plant Sci. 2007 Dec;12(12):548-55. Epub 2007 Nov 19.

Transgenic strategies for the nutritional enhancement of plants.

Zhu C, Naqvi S, Gomez-Galera S, Pelacho AM, Capell T, Christou P.

Universitat de Lleida, Av. Alcalde Rovira Roure, 191, E-25198 Lleida, Spain.

The nutrients in the human diet ultimately come from plants. However, all our
major food crops lack certain essential vitamins and minerals. Although a varied
diet provides adequate nutrition, much of the human population, particularly in
developing countries, relies on staple crops, such as rice or maize, which does
not provide the full complement of essential nutrients. Malnutrition is a
significant public health issue in most of the developing world. One way to
address this problem is through the enhancement of staple crops to increase their
essential nutrient content. Here, we review the current strategies for the
biofortification of crops, including mineral fertilization and conventional
breeding but focusing on transgenic approaches which offer the most rapid way to
develop high-nutrient commercial cultivars.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 18006362 [PubMed - indexed for MEDLINE]

193: Vet Immunol Immunopathol. 2008 Jan 15;121(1-2):83-90. Epub 2007 Aug 25.

Foliar extracts from transgenic tomato plants expressing the structural
polyprotein, P1-2A, and protease, 3C, from foot-and-mouth disease virus elicit a
protective response in guinea pigs.

Pan L, Zhang Y, Wang Y, Wang B, Wang W, Fang Y, Jiang S, Lv J, Wang W, Sun Y, Xie
Q.

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
Research Institute, Chinese Academy of Agriculture Science, Xujiaping 11,
Lanzhou, Gansu 730046, PR China. zhangyg@public.lz.gs.cn

The expression of recombinant antigens in transgenic plants is increasingly used
as an alternative method of producing experimental immunogens. In this report, we
describe the production of transgenic tomato plants that express the structural
polyprotein, P1-2A, and protease, 3C, from foot-and-mouth disease (FMDV). P1-2A3C
was inserted into the plant binary vector, pBin438, and transformed into tomato
plants using Agrobacterium tumefaciens strain, GV3101. The presence of P1-2A3C
was confirmed by PCR, transcription was verified by RT-PCR, and recombinant
protein expression was confirmed by sandwich-ELISA and Western blot analyses.
Guinea pigs immunized intramuscularly with foliar extracts from
P1-2A3C-transgenic tomato plants were found to develop a virus-specific antibody
response against FMDV. Vaccinated guinea pigs were fully protected against a
challenge infection, while guinea pigs injected with untransformed plant extracts
failed to elicit an antibody response and were not protected against challenge.
These results demonstrate that transgenic tomato plants expressing the FMDV
structural polyprotein, P1-2A, and the protease, 3C, can be used as a source of
recombinant antigen for vaccine production.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18006078 [PubMed - indexed for MEDLINE]

194: Environ Biosafety Res. 2007 Jul-Sep;6(3):167-81. Epub 2007 Jul 6.

Long term evaluation of field-released genetically modified rhizobia.

Corich V, Giacomini A, Vendramin E, Vian P, Carlot M, Concheri G, Polone E,
Casella S, Nuti MP, Squartini A.

Dipartimento di Biotecnologie Agrarie, Universitö  di Padova, Viale
dell'Universitö  16, 35020 Legnaro, Padova, Italy.

This is the report of the first open field release of genetically modified
microorganisms (GMMs) in Italy. It covers ten years of monitoring, and follows
in-field GMM dynamics from strain release to disappearance below detection
limits, as well as assessment of impact on resident microorganisms. The bacteria
released belong to the nitrogen fixing legume endosymbiont Rhizobium
leguminosarum bv. viciae, and were engineered with non-agronomically-proficient
traits, in order to assess their behavior and fate without GMM-specific positive
feedback from the plant. A DNA cassette containing mercury resistance and
ss-galactosidase genes was introduced in either plasmid-borne or chromosomally
integrated versions, in order to test the resulting strain stability. A synthetic
promoter was used to drive the lacZ gene, conferring high catabolic activity to
the GMM. Two different wild-type Rhizobium backgrounds were tested, comparing a
non-indigenous vs. an indigenous, highly competitive strain. The latter had much
greater persistence, since it was able to survive and establish at technically
detectable levels for over four years after release. Selection factors, such as
reiterated presence of the plant host, or lactose substrate supply, enhanced
long-term survival to different extents. The lactose treatment showed that even a
single trophic supplementation can surpass the benefits of symbiotic interaction
for a period of several years. Concerning impact, the GMMs did not alter
substantially the other soil community general microbiota. However, there were
some significant differences in microbiota as a consequence of the Rhizobium
inoculation. This effect was observed with either the WT or GMM, and was more
evident in the release of the indigenous Rhizobium. Moreover, as the indigenous
GMM had its parental, dominant wild-type in the same soil, it was possible to
evaluate to what extent the GMM version could result in parent displacement
("self-impact"), and how much the two rhizobia would additively contribute to
nodulation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 18001684 [PubMed - indexed for MEDLINE]

195: Plant Cell Environ. 2008 Jan;31(1):165-76. Epub 2007 Nov 12.

RNA interference-mediated repression of sucrose-phosphatase in transgenic potato
tubers (Solanum tuberosum) strongly affects the hexose-to-sucrose ratio upon cold
storage with only minor effects on total soluble carbohydrate accumulation.

Chen S, Hajirezaei MR, Zanor MI, Hornyik C, Debast S, Lacomme C, Fernie AR,
Sonnewald U, Bö¶rnke F.

Friedrich-Alexander-Universitö¤t, Lehrstuhl fö¼r Biochemie, Staudtstr. 5, 91058
Erlangen, Germany.

Storage of potato tubers at low temperatures leads to the accumulation of glucose
and fructose in a process called 'cold sweetening'. The aim of this work was to
investigate the role of sucrose-phosphatase (SPP) in potato tuber carbohydrate
metabolism at low temperature (4 degrees C). To this end, RNA interference (RNAi)
was used to reduce SPP expression in transgenic potato tubers. Analysis of SPP
specific small interfering RNAs (siRNAs), SPP protein accumulation and enzyme
activity indicated that SPP silencing in transgenic tubers was stable during the
cold treatment. Analysis of soluble carbohydrates showed that in transgenic
tubers, cold-induced hexogenesis was inhibited while, despite strongly reduced
SPP activity, sucrose levels exceeded wild-type (WT) values four- to fivefold
after 34 d of cold treatment. This led to a drastic change in the
hexose-to-sucrose ratio from 1.9 in WT tubers to 0.15 to 0.11 in transgenic
tubers, while the total amount of soluble sugars was largely unchanged in both
genotypes. Sucrose-6(F)-phosphate (Suc6P), the substrate of SPP, accumulated in
transgenic tubers in the cold which most likely enables the residual enzyme to
operate with maximal catalytic activity in vivo and thus, in the long term,
counterbalances reduced SPP activity in the transformants. Northern analysis
revealed that cold-induced expression of vacuolar invertase (VI) was blocked in
SPP-silenced tubers explaining a reduced sucrose-to-hexose conversion. Suc6P
levels were found to negatively correlate with VI expression. A possible role of
Suc6P in regulating VI expression is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17999659 [PubMed - indexed for MEDLINE]

196: J Agric Food Chem. 2007 Dec 12;55(25):10226-31. Epub 2007 Nov 13.

Quantification and persistence of recombinant DNA of Roundup Ready corn and
soybean in rotation.

Lerat S, Gulden RH, Hart MM, Powell JR, England LS, Pauls KP, Swanton CJ,
Klironomos JN, Trevors JT.

Departments of Environmental Biology, Plant Agriculture, and Integrative Biology,
University of Guelph, Guelph, Ontario, Canada N1G 2W1.

The presence of the recombinant cp4 epsps gene from Roundup Ready (RR) corn and
RR soybean was quantified using real-time PCR in soil samples from a field
experiment growing RR and conventional corn and soybean in rotation. RR corn and
RR soybean cp4 epsps persisted in soil for up to 1 year after seeding. The
concentration of recombinant DNA in soil peaked in July and August in RR corn and
RR soybean plots, respectively. A small fraction of soil samples from plots
seeded with conventional crops contained recombinant DNA, suggesting transgene
dispersal by means of natural process or agricultural practices. This research
will aid in the understanding of the persistence of recombinant DNA in
agricultural cropping systems.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17997522 [PubMed - indexed for MEDLINE]

197: J Food Sci. 2007 Aug;72(6):S420-4.

Comparison of nutritional quality between Chinese indica rice with sck and cry1Ac
genes and its nontransgenic counterpart.

Li X, Huang K, He X, Zhu B, Liang Z, Li H, Luo Y.

College of Food Science and Nutritional Engineering, China Agricultural
University, Beijing 100083, China.

Nutritional assessment of transgenic crops used for human food and animal feed is
an important aspect of safety evaluations. An insect-resistant rice (IRR) was
generated by the stable insertion of sck, a modified cowpea trypsin inhibitor
gene, and cry1Ac, encoding a crystal protein from Bacillus thuringiensis into the
genome of a common variety of Chinese indica rice. The composition of the brown
and milled rice grain from the resulted IRR line designated Liangyou Kefeng No. 6
was compared with that of the parental rice cultivar Liangyou 2186. Nutrients,
including the proximates, amino acids, fatty acids, minerals, and vitamins, were
measured. The antinutritive components such as phytic acid, lectin, and trypsin
inhibitors were also examined. The data demonstrated that the nutritional quality
of both the brown and milled rice grains from the transgenic line was
substantially equivalent to that of the nontransgenic counterpart, and measured
amounts of nutritional components fell within the range of values reported for
other commercial lines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17995700 [PubMed - indexed for MEDLINE]

198: Anal Bioanal Chem. 2008 Jan;390(1):377-87. Epub 2007 Nov 11.

Detection and characterization of recombinant DNA expressing vip3A-type
insecticidal gene in GMOs--standard single, multiplex and construct-specific PCR
assays.

Singh CK, Ojha A, Bhatanagar RK, Kachru DN.

Industrial Toxicology Research Centre, Post Box No. 80, M. G. Marg, Lucknow,
226001, Uttar Pradesh, India.

Vegetative insecticidal protein (Vip), a unique class of insecticidal protein, is
now part of transgenic plants for conferring resistance against lepidopteron
pests. In order to address the imminent regulatory need for detection and
labeling of vip3A carrying genetically modified (GM) products, we have developed
a standard single PCR and a multiplex PCR assay. As far as we are aware, this is
the first report on PCR-based detection of a vip3A-type gene (vip-s) in
transgenic cotton and tobacco. Our assay involves amplification of a 284-bp
region of the vip-s gene. This assay can possibly detect as many as 20 natural
wild-type isolates bearing a vip3A-like gene and two synthetic genes of vip3A in
transgenic plants. The limit of detection as established by our assay for GM
trait (vip-s) is 0.1%. Spiking with nontarget DNA originating from diverse plant
sources had no inhibitory effect on vip-s detection. Since autoclaving of vip-s
bearing GM leaf samples showed no deterioration/interference in detection
efficacy, the assay seems to be suitable for processed food products as well. The
vip-s amplicon identity was reconfirmed by restriction endonuclease assay. The
primer set for vip-s was equally effective in a multiplex PCR assay format
(duplex, triplex and quadruplex), used in conjunction with the primer sets for
the npt-II selectable marker gene, Cauliflower mosaic virus 35S promoter and
nopaline synthetase terminator, enabling concurrent detection of the transgene,
regulatory sequences and marker gene. Further, the entire transgene construct was
amplified using the forward primer of the promoter and the reverse primer of the
terminator. The resultant amplicon served as a template for nested PCR to confirm
the construct integrity. The method is suitable for screening any vip3A-carrying
GM plant and food. The availability of a reliable PCR assay method prior to
commercial release of vip3A-based transgenic crops and food would facilitate
rapid and efficient regulatory compliance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17994293 [PubMed - indexed for MEDLINE]

199: Plant Physiol. 2008 Jan;146(1):162-77. Epub 2007 Nov 9.

Involvement of polyamine oxidase in wound healing.

Angelini R, Tisi A, Rea G, Chen MM, Botta M, Federico R, Cona A.

Dipartimento di Biologia, Universitö  Roma Tre, Viale G Marconi 446, 00146, Rome,
Italy.

Hydrogen peroxide (H(2)O(2)) is involved in plant defense responses that follow
mechanical damage, such as those that occur during herbivore or insect attacks,
as well as pathogen attack. H(2)O(2) accumulation is induced during wound healing
processes as well as by treatment with the wound signal jasmonic acid. Plant
polyamine oxidases (PAOs) are H(2)O(2) producing enzymes supposedly involved in
cell wall differentiation processes and defense responses. Maize (Zea mays) PAO
(ZmPAO) is a developmentally regulated flavoprotein abundant in primary and
secondary cell walls of several tissues. In this study, we investigated the
effect of wounding on ZmPAO gene expression in the outer tissues of the maize
mesocotyl and provide evidence that ZmPAO enzyme activity, protein, and mRNA
levels increased in response to wounding as well as jasmonic acid treatment.
Histochemically detected ZmPAO activity especially intensified in the epidermis
and in the wound periderm, suggesting a tissue-specific involvement of ZmPAO in
wound healing. The role played by ZmPAO-derived H(2)O(2) production in
peroxidase-mediated wall stiffening events was further investigated by exploiting
the in vivo use of N-prenylagmatine (G3), a selective and powerful ZmPAO
inhibitor, representing a reliable diagnostic tool in discriminating
ZmPAO-mediated H(2)O(2) production from that generated by peroxidase, oxalate
oxidase, or by NADPH oxidase activity. Here, we demonstrate that G3 inhibits
wound-induced H(2)O(2) production and strongly reduces lignin and suberin
polyphenolic domain deposition along the wound, while it is ineffective in
inhibiting the deposition of suberin aliphatic domain. Moreover, ZmPAO ectopic
expression in the cell wall of transgenic tobacco (Nicotiana tabacum) plants
strongly enhanced lignosuberization along the wound periderm, providing evidence
for a causal relationship between PAO and peroxidase-mediated events during wound
healing.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17993545 [PubMed - indexed for MEDLINE]

200: Plant Physiol. 2008 Jan;146(1):189-99. Epub 2007 Nov 9.

A versatile transposon-based activation tag vector system for functional genomics
in cereals and other monocot plants.

Qu S, Desai A, Wing R, Sundaresan V.

Section of Plant Biology , University of California, Davis, CA 95616, USA.

Transposon insertional mutagenesis is an effective alternative to T-DNA
mutagenesis when transformation through tissue culture is inefficient as is the
case for many crop species. When used as activation tags, transposons can be
exploited to generate novel gain-of-function phenotypes without transformation
and are of particular value in the study of polyploid plants where gene knockouts
will not have phenotypes. We have developed an in cis-activation-tagging Ac-Ds
transposon system in which a T-DNA vector carries a Dissociation (Ds) element
containing 4x cauliflower mosaic virus enhancers along with the Activator (Ac)
transposase gene. Stable Ds insertions were selected using green fluorescent
protein and red fluorescent protein genes driven by promoters that are functional
in maize (Zea mays) and rice (Oryza sativa). The system has been tested in rice,
where 638 stable Ds insertions were selected from an initial set of 26 primary
transformants. By analysis of 311 flanking sequences mapped to the rice genome,
we could demonstrate the wide distribution of the elements over the rice
chromosomes. Enhanced expression of rice genes adjacent to Ds insertions was
detected in the insertion lines using semiquantitative reverse transcription-PCR
method. The in cis-two-element vector system requires minimal number of primary
transformants and eliminates the need for crossing, while the use of fluorescent
markers instead of antibiotic or herbicide resistance increases the applicability
to other plants and eliminates problems with escapes. Because Ac-Ds has been
shown to transpose widely in the plant kingdom, the activation vector system
developed in this study should be of utility more generally to other monocots.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17993541 [PubMed - indexed for MEDLINE]

201: Nat Biotechnol. 2007 Nov;25(11):1213; discussion 1213.

Comment on:
Nat Biotechnol. 2007 Sep;25(9):950.

Open season?

Moses V.

Publication Types:
Comment

PMID: 17989668 [PubMed - indexed for MEDLINE]

202: Nat Biotechnol. 2007 Nov;25(11):1213-4.

GMO quantification in processed food and feed.

Weighardt F.

Publication Types:
Letter

PMID: 17989666 [PubMed - indexed for MEDLINE]

203: Toxicol Sci. 2008 Feb;101(2):215-25. Epub 2007 Nov 7.

Acrylamide-responsive genes in the nematode Caenorhabditis elegans.

Hasegawa K, Miwa S, Isomura K, Tsutsumiuchi K, Taniguchi H, Miwa J.

Institute for Biological Function, Chubu University, Kasugai 487-8501, Japan.

As acrylamide is a known neurotoxin for many animals and potential carcinogen for
humans, it came as a surprise when the Swedish National Food Agency and Stockholm
University reported in 2002 that it is formed during the frying or baking of
foods. We report here genomic and proteomic analyses on genes and proteins of
Caenorhabditis elegans exposed to 500 mg/l acrylamide. Of the 21,120 genes
profiled, 409 genes were more than twofold upregulated and 111 genes were
downregulated. Upregulated genes included many that encode detoxification enzymes
such as glutathione S-transferases (GSTs), uridine
diphosphate-glucuronosyl/glucosyl transferases, and short-chain type
dehydrogenases but only one cytochrome P450. Subsequent proteomic analysis
confirmed the heavy involvement of GSTs. Because of their high expression levels
and central roles in acrylamide metabolism, we analyzed the in vivo expression
patterns of eight gst genes. Although all encoded GST and were more than twofold
upregulated by acrylamide treatment, their expression patterns were varied, and
their regulation involved the transcription factor SKN-1 (a C. elegans homolog of
Nuclear factor E2-related factors 1 and 2). We then selected the
gst-4::gfp-transformed C. elegans to study the detoxification rate of acrylamide
and its metabolite glycidimide in living animals. This animal detects acrylamide
as a green fluorescence protein (GFP) expression signal in a dose- and
time-dependent manner and may prove to be a useful tool not only for rapidly and
inexpensively detecting acrylamide, a harmful substance in food, but also for
analyzing mechanisms of GST induction by acrylamide and other inducers like
oxidative stresses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17989133 [PubMed - indexed for MEDLINE]

204: Biochem Biophys Res Commun. 2008 Jan 11;365(2):334-9. Epub 2007 Nov 5.

Generation of a transgenic rice seed-based edible vaccine against house dust mite
allergy.

Yang L, Kajiura H, Suzuki K, Hirose S, Fujiyama K, Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.

As an alternative approach to conventional allergen-specific immunotherapy,
transgenic rice seed expressing a major house dust mite (HDM) allergen, Der p 1,
was developed as an edible vaccine. The C-terminal KDEL-tagged Der p 1 allergen
specifically accumulated in seed endosperm tissue under the control of the
endosperm-specific GluB1 promoter. Der p 1 reached a maximum concentration of 58
microg/grain and was deposited in the endoplasmic reticulum (ER)-derived protein
body I (PB-I). Plant-derived Der p 1 was posttranslationally modified with
high-mannose-type glycan structures. Glycosylated Der p 1 displayed reduced IgE
binding capacity in comparison with its unglycosylated counterpart in vitro. Our
results indicate that transgenic Der p 1 rice seeds are a safe, potential oral
delivery vaccine for the treatment of HDM allergy.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17988639 [PubMed - indexed for MEDLINE]

205: Med Pregl. 2007 May-Jun;60(5-6):295-8.

[Diseases caused by viruses and toxins in biological warfare and bioterrorism]

[Article in Serbian]

Bojiä‡ I, Vukadinov J, Miniä‡ S.

Specijalisticka ordinacija Dr Bojiä‡ Beograd. drbojic@net.yu

INTRODUCTION: Viruses and toxins, as well as bacteria and rickettsia can
potentially be used as biological weapons in conflicts or in bioterrorism. USE OF
BIOLOGICAL WEAPONS: The infection can be acquired by inhalation of aerosols,
ingestion of contaminated food or water, or direct contact with the skin or
mucosa. Special attention must be given to the possible use of genetically
modified agents. CONCLUSION: This paper describes the clinical features of
diseases caused hbi viruses (smallpox, hemorrhagic Jever and encephalitis) and
toxins (botulinum, staphylococcal enterotoxin B, ricinus toxin and mycotoxins)
their diagnosis, treatment, as well as basic preventive measures.

Publication Types:
English Abstract
Review

PMID: 17988067 [PubMed - indexed for MEDLINE]

206: Crit Rev Food Sci Nutr. 2007;47(8):721-33.

Toxicity studies of genetically modified plants: a review of the published
literature.

Domingo JL.

Laboratory of Toxicology and Environmental Health, School of Medicine, Rovira I
Virgili University, San Lorenzo, Reus, Spain. joseluis.domingo@urv.cat

According to the information reported by the WHO, the genetically modified (GM)
products that are currently on the international market have all passed risk
assessments conducted by national authorities. These assessments have not
indicated any risk to human health. In spite of this clear statement, it is quite
amazing to note that the review articles published in international scientific
journals during the current decade did not find, or the number was particularly
small, references concerning human and animal toxicological/health risks studies
on GM foods. In this paper, the scientific information concerning the potential
toxicity of GM/transgenic plants using the Medline database is reviewed. Studies
about the safety of the potential use of potatoes, corn, soybeans, rice,
cucumber, tomatoes, sweet pepper, peas, and canola plants for food and feed were
included. The number of references was surprisingly limited. Moreover, most
published studies were not performed by the biotechnology companies that produce
these products. This review can be concluded raising the following question:
where is the scientific evidence showing that GM plants/food are toxicologically
safe?

Publication Types:
Review

PMID: 17987446 [PubMed - indexed for MEDLINE]

207: Biosci Biotechnol Biochem. 2007 Nov;71(11):2759-65. Epub 2007 Nov 7.

Chloroplast-like organelles were found in enucleate sieve elements of transgenic
plants overexpressing a proteinase inhibitor.

Xie J, Ouyang XZ, Xia KF, Huang YF, Pan WB, Cai YP, Xu X, Li B, Xu ZF.

State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the
Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou
510275, China.

SaPIN2a, a plant proteinase inhibitor from nightshade (Solanum americanum), was
located to the enucleate sieve elements (SEs) of phloem. The expressed SaPIN2a in
transgenic lettuce showed inhibition of plant endogenous trypsin- and
chymotrypsin-like activities, suggesting that SaPIN2a can regulate proteolysis in
plant cells. To further investigate the physiological role of SaPIN2a, we
produced transgenic nightshade and lettuce plants overexpressing SaPIN2a from the
cauliflower mosaic virus (CaMV) 35S promoter using Agrobacterium-mediated
transformation. Overexpression of SaPIN2a in transgenic plants was demonstrated
by northern blot and western blot analysis. SaPIN2a-overexpressing transgenic
nightshade plants showed significantly lower height than wild-type plants.
Transmission electron microscopy analysis showed that chloroplast-like organelles
with thylakoids, which are not present in enucleate SEs of wild-type plants, were
present in the enucleate SEs of SaPIN2a-overexpressing transgenic plants. This
finding is discussed in terms of the possible role played by SaPIN2a in the
regulation of proteolysis in SEs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17986772 [PubMed - indexed for MEDLINE]

208: Plant Cell Environ. 2008 Jan;31(1):86-96. Epub 2007 Nov 6.

Identification of novel pathogen-responsive cis-elements and their binding
proteins in the promoter of OsWRKY13, a gene regulating rice disease resistance.

Cai M, Qiu D, Yuan T, Ding X, Li H, Duan L, Xu C, Li X, Wang S.

National Key Laboratory of Crop Genetic Improvement, National Center of Plant
Gene Research (Wuhan), Huazhong Agricultural University, Wuhan 430070, China.

The WRKY transcription factor superfamily controls diverse developmental and
physiological processes in plants. However, little is known about the factors
that directly regulate the function of WRKY genes. In this study, we identified
cis-acting elements and their binding proteins of rice OsWRKY13, a gene that
plays a pivotal role in disease resistance against bacterial and fungal
pathogens. Two novel pathogen-responsive cis-elements, PRE2 and PRE4, were
characterized from the promoter region of OsWRKY13. The two cis-elements
negatively regulate gene expression without pathogen challenge, and positively
regulate gene expression after pathogen-induced protein binding. OsWRKY13 binds
to PRE4, which harbours a novel W-like box. Another five proteins (Rad51-like;
tubby-like; SWIM zinc finger and nucleotide-binding adaptor shared by APAF-1,
certain R proteins and CED-4 (NB-ARC) domain containing proteins; and an unknown
protein) also bind to one of the two cis-elements. Different proteins interacting
with the same cis-element appear to have different DNA-binding core sequences.
These proteins localize in the nucleus and show differential expression upon
pathogen challenge. These results suggest that OsWRKY13 expression is regulated
by multiple factors to achieve disease resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17986178 [PubMed - indexed for MEDLINE]

209: Transgenic Res. 2007 Dec;16(6):713-21. Epub 2007 Feb 16.

Biologically active human GM-CSF produced in the seeds of transgenic rice plants.

Sardana R, Dudani AK, Tackaberry E, Alli Z, Porter S, Rowlandson K, Ganz P,
Altosaar I.

Department of Biochemistry, Microbiology & Immunology, Faculty of Medicine,
University of Ottawa, 451 Smyth Road, Ottawa, ON, Canada, K1H 8M5.

Rice flour is a well-known and characterized source of pharmaceutical
ingredients, which are gluten-free and incorporated in many drug delivery
applications such as excipient starch. To further exploit this uniqueness, the
synthetic capacity of rice endosperm tissue, the basis of rice flour, was
extended by genetic transformation. Recombinant human GM-CSF, a cytokine used in
treating neutropenia and with other potential clinical applications, has been
expressed in transgenic rice seeds using a rice glutelin promoter. Rice seeds
accumulated human GM-CSF to a level of 1.3% of total soluble protein. The rice
seed-produced human GM-CSF was found to be biologically active when tested using
a human cell line TF-1. Use of rice as a host plant offers not only attractive
features of safe production in seeds but also self-containment of foreign genes,
as rice is primarily a self-pollinated crop plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17985214 [PubMed - indexed for MEDLINE]

210: Regul Toxicol Pharmacol. 2008 Feb;50(1):98-113. Epub 2007 Sep 29.

Comparative safety assessment of plant-derived foods.

Kok EJ, Keijer J, Kleter GA, Kuiper HA.

RIKILT Institute of Food Safety, Bornsesteeg 45, PO Box 230, 6700 AE Wageningen,
The Netherlands. Esther.kok@wur.nl

The second generation of genetically modified (GM) plants that are moving towards
the market are characterized by modifications that may be more complex and traits
that more often are to the benefit of the consumer. These developments will have
implications for the safety assessment of the resulting plant products. In part
of the cases the same crop plant can, however, also be obtained by 'conventional'
breeding strategies. The breeder will decide on a case-by-case basis what will be
the best strategy to reach the set target and whether genetic modification will
form part of this strategy. This article discusses important aspects of the
safety assessment of complex products derived from newly bred plant varieties
obtained by different breeding strategies. On the basis of this overview, we
conclude that the current process of the safety evaluation of GM versus
conventionally bred plants is not well balanced. GM varieties are elaborately
assessed, yet at the same time other crop plants resulting from conventional
breeding strategies may warrant further food safety assessment for the benefit of
the consumer. We propose to develop a general screening frame for all newly
developed plant varieties to select varieties that cannot, on the basis of
scientific criteria, be considered as safe as plant varieties that are already on
the market.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't
Review

PMID: 17983697 [PubMed - indexed for MEDLINE]

211: Nat Biotechnol. 2007 Nov;25(11):1322-6. Epub 2007 Nov 4.

Control of coleopteran insect pests through RNA interference.

Baum JA, Bogaert T, Clinton W, Heck GR, Feldmann P, Ilagan O, Johnson S,
Plaetinck G, Munyikwa T, Pleau M, Vaughn T, Roberts J.

Monsanto Company, 700 Chesterfield Parkway West, Chesterfield, Missouri
63017-1732, USA.

Commercial biotechnology solutions for controlling lepidopteran and coleopteran
insect pests on crops depend on the expression of Bacillus thuringiensis
insecticidal proteins, most of which permeabilize the membranes of gut epithelial
cells of susceptible insects. However, insect control strategies involving a
different mode of action would be valuable for managing the emergence of insect
resistance. Toward this end, we demonstrate that ingestion of double-stranded
(ds)RNAs supplied in an artificial diet triggers RNA interference in several
coleopteran species, most notably the western corn rootworm (WCR) Diabrotica
virgifera virgifera LeConte. This may result in larval stunting and mortality.
Transgenic corn plants engineered to express WCR dsRNAs show a significant
reduction in WCR feeding damage in a growth chamber assay, suggesting that the
RNAi pathway can be exploited to control insect pests via in planta expression of
a dsRNA.

PMID: 17982443 [PubMed - indexed for MEDLINE]

212: Plant Physiol. 2008 Jan;146(1):213-27. Epub 2007 Nov 2.

Glufosinate ammonium-induced pathogen inhibition and defense responses culminate
in disease protection in bar-transgenic rice.

Ahn IP.

National Institute of Agricultural Biotechnology, Rural Development
Administration, Suwon, Korea. jinhyung@rda.go.kr

Glufosinate ammonium diminished developments of rice (Oryza sativa) blast and
brown leaf spot in 35S:bar-transgenic rice. Pre- and postinoculation treatments
of this herbicide reduced disease development. Glufosinate ammonium specifically
impeded appressorium formation of the pathogens Magnaporthe grisea and
Cochliobolus miyabeanus on hydrophobic surface and on transgenic rice. In
contrast, conidial germination remained unaffected. Glufosinate ammonium
diminished mycelial growth of two pathogens; however, this inhibitory effect was
attenuated in malnutrition conditions. Glufosinate ammonium caused slight
chlorosis and diminished chlorophyll content; however, these alterations were
almost completely restored in transgenic rice within 7 d. Glufosinate ammonium
triggered transcriptions of PATHOGENESIS-RELATED (PR) genes and hydrogen peroxide
accumulation in transgenic rice and PR1 transcription in Arabidopsis (Arabidopsis
thaliana) wild-type ecotype Columbia harboring 35S:bar construct. All transgenic
Arabidopsis showed robust hydrogen peroxide accumulation by glufosinate ammonium.
This herbicide also induced PR1 transcription in etr1 and jar1 expressing bar;
however, no expression was observed in NahG and npr1. Fungal infection did not
alter transcriptions of PR genes and hydrogen peroxide accumulation induced by
glufosinate ammonium. Infiltration of glufosinate ammonium did not affect
appressorium formation of M. grisea in vivo but inhibited blast disease
development. Hydrogen peroxide scavengers nullified blast protection and
transcriptions of PR genes by glufosinate ammonium; however, they did not affect
brown leaf spot progression. In sum, both direct inhibition of pathogen infection
and activation of defense systems were responsible for disease protection in
bar-transgenic rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17981989 [PubMed - indexed for MEDLINE]

213: Plant Physiol. 2007 Dec;145(4):1192-200. Epub 2007 Nov 2.

A set of modular binary vectors for transformation of cereals.

Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer P, Kumlehn J.

Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben,
Germany.

Genetic transformation of crop plants offers the possibility of testing
hypotheses about the function of individual genes as well as the exploitation of
transgenes for targeted trait improvement. However, in most cereals, this option
has long been compromised by tedious and low-efficiency transformation protocols,
as well as by the lack of versatile vector systems. After having adopted and
further improved the protocols for Agrobacterium-mediated stable transformation
of barley (Hordeum vulgare) and wheat (Triticum aestivum), we now present a
versatile set of binary vectors for transgene overexpression, as well as for gene
silencing by double-stranded RNA interference. The vector set is offered with a
series of functionally validated promoters and allows for rapid integration of
the desired genes or gene fragments by GATEWAY-based recombination. Additional
in-built flexibility lies in the choice of plant selectable markers, cassette
orientation, and simple integration of further promoters to drive specific
expression of genes of interest. Functionality of the cereal vector set has been
demonstrated by transient as well as stable transformation experiments for
transgene overexpression, as well as for targeted gene silencing in barley.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17981986 [PubMed - indexed for MEDLINE]

214: Mol Cells. 2007 Oct 31;24(2):301-6.

Overexpression of Arabidopsis homogentisate phytyltransferase or tocopherol
cyclase elevates vitamin E content by increasing gamma-tocopherol level in
lettuce (Lactuca sativa L.).

Lee K, Lee SM, Park SR, Jung J, Moon JK, Cheong JJ, Kim M.

School of Agricultural Biotechnology and Center for Agricultural Biomaterials,
Seoul National University, Seoul 151-921, Korea.

Tocopherols, essential components of the human diet, are synthesized exclusively
by photosynthetic organisms. To increase tocopherol content by increasing total
flux to the tocopherol biosynthetic pathway, genes encoding Arabidopsis
homogentisate phytyltransferase (HPT/V-TE2) and tocopherol cyclase (TC/VTE1) were
constitutively overexpressed in lettuce (Lactuca sativa L.). Total tocopherol
content of the transgenic plants overexpressing either of the genes was increased
by more than 2-fold mainly due to an increase in gamma-tocopherol. However,
chlorophyll content in the HPT/VTE2 and TC/VTE1 transgenic lines decreased by up
to 20% and increased by up to 35%, respectively (P < 0.01). These results
demonstrate that manipulation of the tocopherol biosynthetic pathway can increase
or decrease chlorophyll content depending on the gene introduced.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17978586 [PubMed - indexed for MEDLINE]

215: Mol Plant Microbe Interact. 2007 Nov;20(11):1346-52.

Salicylic acid is important for basal defense of Solanum tuberosum against
Phytophthora infestans.

Halim VA, Eschen-Lippold L, Altmann S, Birschwilks M, Scheel D, Rosahl S.

Leibniz Institute of Plant Biochemistry, Department of Stress and Developmental
Biology, Weinberg 3, D-06120 Halle (Saale), Germany.

The importance of the signaling compound salicylic acid for basal defense of
potato (Solanum tuberosum L. cv. Dö©sirö©e) against Phytophthora infestans, the
causal agent of late blight disease, was assessed using transgenic NahG potato
plants which are unable to accumulate salicylic acid. Although the size of
lesions caused by P. infestans was not significantly different in wild-type and
transgenic NahG plants, real-time polymerase chain reaction analyses revealed a
drastic enhancement of pathogen growth in potato plants depleted of salicylic
acid. Increased susceptibility of NahG plants correlated with compromised callose
formation and reduced early defense gene expression. NahG plants pretreated with
the salicylic acid analog 2,6-dichloro-isonicotinic acid allowed pathogen growth
to a similar extent as did wild-type plants, indicating that salicylic acid is an
important compound required for basal defense of potato against P. infestans.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17977146 [PubMed - indexed for MEDLINE]

216: J Am Vet Med Assoc. 2007 Nov 1;231(9):1340-2.

Comment in:
J Am Vet Med Assoc. 2008 Feb 1;232(3):350; author reply 350-1.

Envisioning the future of veterinary medicine: the imperative for change in
veterinary medical education.

Prasse KW, Heider LE, Maccabe AT.

Association of American Veterinary Medical Colleges, 1101 Vermont Ave NW, Ste
301, Washington, DC 20005, USA.

PMID: 17975989 [PubMed - indexed for MEDLINE]

217: Science. 2007 Dec 7;318(5856):1561-2. Epub 2007 Nov 1.

Comment on:
Science. 2007 Dec 7;318(5856):1640-2.

Plant science. The power of the pyramid.

Moar WJ, Anilkumar KJ.

Department of Entomology and Plant Pathology, Auburn University, Auburn, AL
36849, USA. moarwil@auburn.edu

Publication Types:
Comment

PMID: 17975032 [PubMed - indexed for MEDLINE]

218: Ecol Appl. 2007 Oct;17(7):2123-35.

Effect of pollinator abundance on self-fertilization and gene flow: application
to GM Canola.

Hoyle M, Hayter K, Cresswell JE.

PenTAG, Peninsula Medical School, University of Plymouth, Noy Scott House,
Barrack Road, Exeter EX2 5DW, United Kingdom. m.w.hoyle@exeter.ac.uk

Cross-pollination from fields of transgenic crops is of great public concern.
Although cross-pollination in commercial canola (Brassica napus) fields has been
empirically measured, field trials are expensive and do not identify the causes
of cross-pollination. Therefore, theoretical models can be valuable because they
can provide estimates of cross-pollination at any given site and time. We present
a general analytical model of field-to-field gene flow due to the following
competing mechanisms: the wind, bees, and autonomous pollination. We parameterize
the model for the particular case of field-to-field cross-pollination of
genetically modified (GM) canola via the wind and via bumble bees (Bombus spp.)
and honey bees (Apis mellifera). We make extensive use of the large data set of
bee densities collected during the recent U.K. Farm Scale Evaluations. We predict
that canola approaches almost full seed set without pollinators and that
autonomous pollination is responsible for > or = 25% of seed set, irrespective of
pollinator abundance. We do not predict the relative contribution of bees vs. the
wind in landscape-scale gene flow in canola. However, under model assumptions, we
predict that the maximum field-to-field gene flow due to bumble bees is 0.04% and
0.13% below the current EU limit for adventitious GM presence for winter- and
spring-sown canola, respectively. We predict that gene flow due to bees is
approximately 3.1 times higher at 20% compared to 100% male-fertility, and due to
the wind, 1.3 times higher at 20% compared to 100% male-fertility, for both
winter- and spring-sown canola. Bumble bee-mediated gene flow is approximately
2.7 times higher and wind-mediated gene flow approximately 1.7 times lower in
spring-sown than in winter-sown canola, regardless of the degree of
male-sterility. The model of cross-pollination due to the wind most closely
predicted three previously published observations: field-to-field gene flow is
low; gene flow increases with the proportion of plants that are male-sterile; and
gene flow is higher in winter- than in spring-sown canola. Our results therefore
suggest that the wind, not bees, is the main vector of long-distance gene flow in
canola.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17974346 [PubMed - indexed for MEDLINE]

219: Proteomics. 2007 Dec;7(23):4349-57.

Effect of the 1BL.1RS translocation on the wheat endosperm, as revealed by
proteomic analysis.

Gobaa S, Bancel E, Kleijer G, Stamp P, Branlard G.

Swiss Federal Institute of Technology Zurich (ETHZ), Institute of Plant Sciences,
Universitö¤tsrasse 2, Switzerland. samy.gobaa@ipw.agrl.ethz.ch

The introduction of the 1RS chromosome of rye into wheat made wheat more
resistant to several pathogens. Today, this resistance has been overcome but the
1BL.1RS translocation remains interesting because of the improved yield and
despite the lower rheological properties it produces. Nothing has been reported
yet on the impact of rye chromatin introgression on the grain proteome of wheat.
The comparison of the 2-DE profiles of 16 doubled haploid lines, with or without
the 1BL.1RS translocation, revealed quantitative and qualitative proteic
variations in prolamins and other endosperm proteins. Eight spots were found
specifically in lines having the 1BL.1RS translocation; 16 other spots
disappeared from the same lines. Twelve spots, present in both genotypes, met the
criteria for up- or down-regulated spots. In translocated genotypes, a highly
overexpressed spot, identified as a gamma-gliadin with nine cysteine residues,
suggests that the lack of LMW-GS induced by 1BL.1RS is counterbalanced by an
overexpression of a relatively similar prolamin. Moreover, a spot that was absent
from 1BL.1RS genotypes was identified as a dimeric alpha-amylase inhibitor. It
was considered to be a valuable candidate to explain the sticky dough associated
with translocated cultivars.

PMID: 17973293 [PubMed - indexed for MEDLINE]

220: Development. 2007 Nov;134(22):4107-17.

Arabidopsis GLAUCE promotes fertilization-independent endosperm development and
expression of paternally inherited alleles.

Ngo QA, Moore JM, Baskar R, Grossniklaus U, Sundaresan V.

Section of Plant Biology, University of California, One Shields Avenue, Davis, CA
95616, USA.

Early seed development of sexually reproducing plants requires both maternal and
paternal genomes but is prominently maternally influenced. A novel gametophytic
maternal-effect mutant defective in early embryo and endosperm development,
glauce (glc), has been isolated from a population of Arabidopsis Ds transposon
insertion lines. The glc mutation results from a deletion at the Ds insertion
site, and the molecular identity of GLC is not known. glc embryos can develop up
to the globular stage in the absence of endosperm and glc central cells appear to
be unfertilized. glc suppresses autonomous endosperm development observed in the
fertilization-independent seed (fis) class mutants. glc is also epistatic to mea,
one of the fis class mutants, in fertilized seeds, and is essential for the
biparental embryonic expression of PHE1, a repressed downstream target of MEA. In
addition, maternal GLC function is required for the paternal embryonic expression
of the ribosome protein gene RPS5a and the AMP deaminase gene FAC1, both of which
are essential for early embryo and endosperm development. These results indicate
that factors derived from the female gametophyte activate a subset of the
paternal genome of fertilized seeds.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17965055 [PubMed - indexed for MEDLINE]

221: Int J Toxicol. 2007 Sep-Oct;26(5):389-99.

Strategies to evaluate the safety of bioengineered foods.

Delaney B.

Pioneer Hi-Bred International, Inc., DuPont Agriculture and Nutrition, Johnston,
Iowa 50131-0550, USA. bryan.delaney@pioneer.com

A number of genetically modified (GM) crops bioengineered to express agronomic
traits including herbicide resistance and insect tolerance have been
commercialized. Safety studies conducted for the whole grains and food and feed
fractions obtained from GM crops (i.e., bioengineered foods) bear similarities to
and distinctive differences from those applied to substances intentionally added
to foods (e.g., food ingredients). Similarities are apparent in common animal
models, route of exposure, duration, and response variables typically assessed in
toxicology studies. However, because of differences in the nutritional and
physical properties of food ingredients and bioengineered foods and in the
fundamental goals of the overall safety assessment strategies for these different
classes of substances, there are recognizable differences in the individual
components of the safety assessment process. The fundamental strategic difference
is that the process for food ingredients is structured toward quantitative risk
assessment whereas that for bioengineered foods is structured for the purpose of
qualitative risk assessment. The strategy for safety assessment of bioengineered
foods focuses on evaluating the safety of the transgenic proteins used to impart
the desired trait or traits and to demonstrate compositional similarity between
the grains of GM and non-GM comparator crops using analytical chemistry and, in
some cases, feeding studies. Despite these differences, the similarities in the
design of safety studies conducted with bioengineered foods should be recognized
by toxicologists. The current paper reviews the basic principles of safety
assessment for bioengineered foods and compares them with the testing strategies
applied to typical food ingredients. From this comparison it can be seen that the
strategies used to assess the safety of bioengineered foods are at least as
robust as that used to assess the safety of typical food ingredients.

Publication Types:
Review

PMID: 17963126 [PubMed - indexed for MEDLINE]

222: J Agric Food Chem. 2007 Nov 28;55(24):9846-9. Epub 2007 Oct 26.

Improving zinc content and antioxidant activity in transgenic tomato plants with
expression of mouse metallothionein-I by mt-I gene.

Sheng J, Liu K, Fan B, Yuan Y, Shen L, Ru B.

College of Food Science, China Agricultural University, Beijing, China.

Metallothioneins (MTs), as a family of low-molecular-weight, cysteine-rich, and
metal-binding proteins, show potential for utilization in functional food. Tomato
plants were transformed with gene constructs that contained mt-I encoding the
mouse MT-I, similar in sense orientation with the constitutively active double
35S promoter from cauliflower mosaic virus. Three independent transformants,
which had copies of the gene in their genomes, were obtained. In these transgenic
lines, high-level expression of MT-I, high zinc content, and some antioxidant
enzyme activities were detected in leaves. The average zinc content in transgenic
tomato leaves was 32.7 mg/100 g FW, which about 1.6 times higher than that in
wild-type. The superoxide dismutase activity was also higher (68.6, 66.9, and
66.1 U/g FW in the three transformants) than that in wild-type (57.4 U/g FW). In
particular, the levels of superoxide free radical scanvenging in the three
transformants were 14.2%, 14.6%, and 13.7%, respectively, which about 1.5 times
higher than that in control (5.6%). Transgenic MT tomato may potentially be used
as an antioxidant and for zinc supplementation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17960876 [PubMed - indexed for MEDLINE]

223: Electrophoresis. 2007 Nov;28(22):4192-201.

CE-MS of zein proteins from conventional and transgenic maize.

Erny GL, Marina ML, Cifuentes A.

Institute of Industrial Fermentations (CSIC), Madrid, Spain.

In this work, an original CE-MS method has been developed to analyze the complex
zein protein fractions from maize. A thorough optimization of: (i) zein protein
extraction, (ii) CE separation, and (iii) electrospray-MS (ESI-MS) detection is
carried out in order to obtain highly informative CE-MS profiles of this
fraction. The developed CE-MS method provides good separation of multiple zein
proteins based on their electrophoretic mobilities as well as adequate
characterization of these proteins based on their M(r). Zein proteins with small
M(r) differences (below 100 Da) were easily separated and successfully analyzed
by CE-MS. Thus, apart of the so-called 15-kDa-beta-zein and 16-kDa-gamma-zein,
which are demonstrated to be formed by a heterogeneous group of proteins,
numerous alpha-zeins belonging to the 19- and 22-kDa fraction were also
identified for the first time in this work. The usefulness of this CE-MS method
was corroborated by comparing the zein-protein fingerprints of various maize
lines including transgenic and their corresponding nontransgenic isogenic lines
cultivated under the same conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17960858 [PubMed - indexed for MEDLINE]

224: Molecules. 2007 Jul 27;12(8):1569-95.

Functional analysis of polyphenol oxidases by antisense/sense technology.

Thipyapong P, Stout MJ, Attajarusit J.

Suranaree University of Technology, 111 University Ave., Muang District, Nakhon
Ratchasima 30000, Thailand. piyada@sut.ac.th

Polyphenol oxidases (PPOs) catalyze the oxidation of phenolics to quinones, the
secondary reactions of which lead to oxidative browning and postharvest losses of
many fruits and vegetables. PPOs are ubiquitous in angiosperms, are inducible by
both biotic and abiotic stresses, and have been implicated in several
physiological processes including plant defense against pathogens and insects,
the Mehler reaction, photoreduction of molecular oxygen by PSI, regulation of
plastidic oxygen levels, aurone biosynthesis and the phenylpropanoid pathway.
Here we review experiments in which the roles of PPO in disease and insect
resistance as well as in the Mehler reaction were investigated using transgenic
tomato (Lycopersicon esculentum) plants with modified PPO expression levels
(suppressed PPO and overexpressing PPO). These transgenic plants showed normal
growth, development and reproduction under laboratory, growth chamber and
greenhouse conditions. Antisense PPO expression dramatically increased
susceptibility while PPO overexpression increased resistance of tomato plants to
Pseudomonas syringae. Similarly, PPO-overexpressing transgenic plants showed an
increase in resistance to various insects, including common cutworm (Spodoptera
litura (F.)), cotton bollworm (Helicoverpa armigera (Hö¼bner)) and beet army worm
(Spodoptera exigua (Hö¼bner)), whereas larvae feeding on plants with suppressed
PPO activity had higher larval growth rates and consumed more foliage. Similar
increases in weight gain, foliage consumption, and survival were also observed
with Colorado potato beetles (Leptinotarsa decemlineata (Say)) feeding on
antisense PPO transgenic tomatoes. The putative defensive mechanisms conferred by
PPO and its interaction with other defense proteins are discussed. In addition,
transgenic plants with suppressed PPO exhibited more favorable water relations
and decreased photoinhibition compared to nontransformed controls and transgenic
plants overexpressing PPO, suggesting that PPO may have a role in the development
of plant water stress and potential for photoinhibition and photooxidative damage
that may be unrelated to any effects on the Mehler reaction. These results
substantiate the defensive role of PPO and suggest that manipulation of PPO
activity in specific tissues has the potential to provide broad-spectrum
resistance simultaneously to both disease and insect pests, however, effects of
PPO on postharvest quality as well as water stress physiology should also be
considered. In addition to the functional analysis of tomato PPO, the application
of antisense/sense technology to decipher the functions of PPO in other plant
species as well as for commercial uses are discussed.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17960074 [PubMed - indexed for MEDLINE]

225: Risk Anal. 2007 Aug;27(4):935-46.

An empirical test of competing theories of hazard-related trust: the case of GM
food.

Allum N.

Department of Sociology, University of Surrey, Guildford, UK.
n.allum@surrey.ac.uk

Few scholars doubt the importance of trust in explaining variation in public
perception of technological risk. Relatively little, however, is known about the
particular types of judgments that people use in granting or withholding trust.
This article presents findings from an empirical study that explores several
dimensions of trust relevant for citizens' judgments of scientists involved in
the development of GM food. The relationship between particular dimensions of
trust and perceptions of GM food risk is also explored, using structural equation
modeling. Results suggest that trust judgments based on the perception of shared
values are most important in relation to GM food risk, but that judgments about
scientists' technical competence are also important.

PMID: 17958502 [PubMed - indexed for MEDLINE]

226: Genetika. 2007 Aug;43(8):1058-64.

[Phytopathological and molecular genetic identification of brown rust resistance
genes in common wheat accessions with alien genetic material]

[Article in Russian]

Gaä­nullin NR, Lapochkina IF, Zhemchuzhina AI, Kiseleva MI, Kolomiets TM,
Kovalenko ED.

Brown rust resistance genes were sought in 23 resistant common wheat accessions
with alien genetic material of Aegilops speltoides, Ae. triuncialis, and Triticum
kiharae from the Arsenal collection. The genes were identified by common
phytopathological tests and PCR analysis with STS markers directed to the known
Lr genes. None of the methods identified the resistance genes in two accessions.
In the other accessions, the combination of the two methods broadened the
spectrum of detectable genes and, in some cases, allowed double verification of
the presence of a resistance gene. Most accessions proved to contain several
brown rust resistance genes, combining juvenile and adult plant ones. The
accessions were found to contain gene combinations that ensured field resistance
and immunity under the conditions of the Non-Chernozem region (Lr13 + Lr10 and
Lr12 + Lr34). Accessions with alien genetic material contained a unique
combination of five or six resistance genes. Since the accessions were rich in
brown rust resistance genes, including effective ones, and carried rare
combinations of these genes, they were proposed as donors to be universally
employed in breeding for immunity in all regions of Russia.

Publication Types:
English Abstract

PMID: 17958305 [PubMed - indexed for MEDLINE]

227: Plant Biotechnol J. 2008 Jan;6(1):2-12. Epub 2007 Oct 23.

US regulatory system for genetically modified [genetically modified organism
(GMO), rDNA or transgenic] crop cultivars.

McHughen A, Smyth S.

Department of Botany and Plant Sciences, University of California, Riverside, CA
92521-0124, USA. alanmc@ucr.edu

This paper reviews the history of the federal regulatory oversight of plant
agricultural biotechnology in the USA, focusing on the scientific and political
forces moulding the continually evolving regulatory structure in place today.
Unlike most other jurisdictions, the USA decided to adapt pre-existing
legislation to encompass products of biotechnology. In so doing, it established
an overarching committee (Office of Science and Technology Policy) to study and
distribute various regulatory responsibilities amongst relevant agencies: the
Food and Drug Administration, Environmental Protection Agency and US Department
of Agriculture. This paper reviews the history and procedures of each agency in
the execution of its regulatory duties and investigates the advantages and
disadvantages of the US regulatory strategy.

Publication Types:
Historical Article
Review

PMID: 17956539 [PubMed - indexed for MEDLINE]

228: J AOAC Int. 2007 Sep-Oct;90(5):1513-6.

Development of agricultural biotechnology and biosafety regulations used to
assess the safety of genetically modified crops in Iran.

Mousavi A, Malboobi MA, Esmailzadeh NS.

National Institute of Genetic Engineering and Biotechnology, P.O. Box 14155-6343,
Tehran, 1417863171, Iran. m-amir@nrcgeb.ac.ir

Rapid progress in the application of biotechnological methodologies and
development of genetically modified crops in Iran necessitated intensive efforts
to establish proper organizations and prepare required rules and regulations at
the national level to ensure safe application of biotechnology in all pertinent
aspects. Practically, preparation of a national biotechnology strategic plan in
the country coincided with development of a national biosafety framework that was
the basis for the drafted biosafety law. Although biosafety measures were
observed by researchers voluntarily, the establishment of national biosafety
organizations since the year 2000 built a great capacity to deal with biosafety
issues in the present and future time, particularly with respect to food and
agricultural biotechnology.

Publication Types:
Review

PMID: 17956001 [PubMed - indexed for MEDLINE]

229: J AOAC Int. 2007 Sep-Oct;90(5):1508-12.

Development of agribiotechnology and biosafety regulations used to assess safety
of genetically modified crops in Bangladesh.

Nasiruddin KM, Nasim A.

Bangladesh Agricultural University, Biotechnology Department, Mymensingh 2202,
Bangladesh. nasirbiotech@yahoo.com

Bangladesh is on the verge of adopting genetically modified (GM) crops for
commercial cultivation and consumption as feed and food. Most of the laboratories
are engaged in tissue culture and molecular characterization on plants, whereas
some have started living modified organism research with shortages of trained
manpower, infrastructure, and funding. Nutritionally improved Golden Rice,
biotech brinjal, and late blight-resistant potato are in contained trials in a
greenhouse, and potato ring spot virus-resistant papaya is in the process of
approval for a field trial. The government has taken some initiative in support
of GM organism research, which include the formation of a Biotechnology
Department in all institutes and the formation of the apex body, the National
Task Force Committee on Biotechnology of Bangladesh under the chairpersonship of
the Prime Minister. Biosafety policy guidelines and related aspects of
biotechnology issues have been approved, and the laws are in the process of being
promulgated. Being a party to the Cartagena Protocol, proper biosafety measures
are regulated by the appropriate authority as stated. Although there are no laws
made yet directly for biosafety of GM crops/foods, the relevant laws on
agriculture, medicine, food, import, trade, environment, etc. may suffice and
explain the situation.

Publication Types:
Review

PMID: 17956000 [PubMed - indexed for MEDLINE]

230: J AOAC Int. 2007 Sep-Oct;90(5):1500-7.

Development of agriculture biotechnology in Pakistan.

Zafar Y.

Pakistan Atomic Energy Commission, Agriculture and Biotechnology Division, PO Box
No. 1114, Islamabad, Pakistan. y_zafar@yahoo.com

Agriculture plays an important role in the national economy of Pakistan, where
most of the rapidly increasing population resides in rural areas and depends on
agriculture for subsistence. Biotechnology has considerable potential for
promoting the efficiency of crop improvement, food production, and poverty
reduction. Use of modern biotechnology started in Pakistan since 1985. Currently,
there are 29 biotech centers/institutes in the country. However, few centers have
appropriate physical facilities and trained manpower to develop genetically
modified (GM) crops. Most of the activities have been on rice and cotton, which
are among the top 5 crops of Pakistan. Biotic (virus/bacterial/insect) and
abiotic (salt) resistant and quality (male sterility) genes have already been
incorporated in some crop plants. Despite acquiring capacity to produce
transgenic plants, no GM crops, either produced locally or imported, have been
released in the country. Pakistan is signatory to the World Trade Organization,
Convention on Biological Diversity, and Cartagena protocols. Several legislations
under the Agreement on Trade-Related Aspects of Intellectual Property Rights have
been promulgated in the country. National Biosafety Guidelines have been
promulgated in April 2005. The Plant Breeders Rights Act, Amendment in Seed
Act-1976, and Geographical Indication for Goods are still passing through
discussion, evaluation, and analysis phases. Meanwhile, an illegal GM crop
(cotton) has already sneaked into farmer's field. Concerted and coordinated
efforts are needed among various ministries for implementation of regulation and
capacity building for import/export and local handling of GM crops. Pakistan
could easily benefit from the experience of Asian countries, especially China and
India, where conditions are similar and the agriculture sector is almost like
that of Pakistan. Thus, the exchange of information and experiences is important
among these nations.

PMID: 17955999 [PubMed - indexed for MEDLINE]

231: J AOAC Int. 2007 Sep-Oct;90(5):1492-9.

Application of current allergy assessment guidelines to next-generation
biotechnology-derived crops.

Bannon GA, Martino-Catt S.

Monsanto Co., Global Regulatory Sciences, 800 N. Lindbergh Blvd, St. Louis, MO
63167, USA. gary.a.bannan@monsanto.com

In any single day, our immune systems are exposed to thousands of different
proteins from the environment and the food we eat. In a portion of the human
population, some of those proteins will stimulate the immune systems to
synthesize immunoglobulin E in an allergenic response. The discrepancy between
the vast numbers of proteins we encounter and the limited number of proteins that
actually become allergens have led scientists on a quest to discover what unique
features exist that make proteins destined to be allergens. The information
gained from these studies has led to an allergy assessment strategy that
characterizes the potential allergenicity of biotechnology products prior to
their commercialization. This testing strategy appears to be effective as shown
by the fact that there have been no clinically documented food allergic reactions
to any of the biotechnology proteins introduced into food crops, to date. The
next generation of biotechnology products will most likely contain more complex
traits, including nutritionally enhanced food crops, and the question arises as
to whether the current allergy assessment strategy will be sufficient to protect
the health of the consuming public. In this paper, we discuss general allergen
characteristics in order to better understand how proteins become allergens,
summarize the current allergy assessment process, evaluate the different aspects
of this process for their adequacy in determining the allergenic potential of
engineered functional foods, and, finally, we assess the possibility of new
technologies having a positive impact on the allergy assessment of nutritionally
enhanced crops.

Publication Types:
Review

PMID: 17955998 [PubMed - indexed for MEDLINE]

232: J AOAC Int. 2007 Sep-Oct;90(5):1480-91.

An overview of methods for assessment of iron bioavailability from foods
nutritionally enhanced through biotechnology.

Cockell KA.

Health Canada, Nutrition Research Division, Food Directorate, 2203C Banting
Research Centre, 251 Sir Frederick Banting Driveway, Ottawa, ON, Canada.
kevin_cockell@hc-sc.gc.ca

Iron deficiency and iron deficiency anemia continue to be significant public
health problems worldwide. While supplementation and fortification have been
viable means to improve iron nutriture of the population in developed countries,
they may be less successful in developing regions for a number of reasons,
including complexities in distribution and consumer compliance. Biofortification
of staple crops, through conventional plant breeding strategies or modern methods
of biotechnology, provides an alternative approach that may be more sustainable
once initial investments have been made. Three types of biofortification
strategies are being essayed, singly or in combination: increasing the total iron
content of edible portions of the plant, decreasing the levels of inhibitors of
iron absorption, and increasing the levels of factors that enhance iron
absorption. Bioavailability is a key concept in iron nutrition, particularly for
nonheme iron such as is found in these biofortified foods. An overview is
presented of methods for evaluation of iron bioavailability from foods
nutritionally enhanced through biotechnology.

PMID: 17955997 [PubMed - indexed for MEDLINE]

233: J AOAC Int. 2007 Sep-Oct;90(5):1470-9.

Nutritional and safety assessments of foods and feeds nutritionally improved
through biotechnology: lysine maize as a case study.

Glenn KC.

Monsanto Co., 800 North Lindbergh Blvd, E3NB, St. Louis, MO 63167, USA.
kevin.c.glenn@monsanto.com

During the last decade, the area of biotech crops modified for agronomic input
traits (e.g., herbicide tolerance and insect protection) has increased to 90
million halyear, grown by over 8 million farmers in a total of 17 countries. As
adoption of these improved agronomic trait biotech crops has grown, so has
interest in biotech crops that have improved nutritional characteristics for use
as feed and food. A previous publication by the International Life Sciences
Institute (ILSI) reported on the principles and concepts proposed for the
nutritional and safety assessments of foods and feeds nutritionally improved
through biotechnology. In this paper, the guidelines and principles recommended
in the earlier publication are discussed relative to a specific case study,
Lysine maize. Lysine maize is a feed ingredient with enhanced nutritional
characteristics for poultry and swine and provides an alternative to the need for
addition of supplemental lysine to some diets for these animals. The 2004 Task
Force of the ILSI has also applied the concepts from that report to 4 other case
studies: sweet potato enriched in provitamin A (2 examples, one using
biotechnology and one using conventional breeding); Golden Rice 2; double-embryo
maize; and ASP-1 enhanced protein sweet potato.

PMID: 17955996 [PubMed - indexed for MEDLINE]

234: J AOAC Int. 2007 Sep-Oct;90(5):1456-64.

Overaccumulation of higher polyamines in ripening transgenic tomato fruit revives
metabolic memory, upregulates anabolism-related genes, and positively impacts
nutritional quality.

Mattoo AK, Chung SH, Goyal RK, Fatima T, Solomos T, Srivastava A, Handa AK.

U.S. Department of Agriculture, Agricultural Research Service, Sustainable
Agricultural Systems Laboratory, The Henry A. Wallace Beltsville Agricultural
Research Center, Bldg 001, Beltsville, MD 20705-2350, USA.
autar.mattoo@ars.usda.gov

Vegetables and fruits are essential components of the human diet as they are
sources of vitamins, minerals, and fiber and provide antioxidants that prevent
chronic diseases. Our goal is to improve durable nutritional quality of tomato
fruit. We developed transgenic tomatoes expressing yeast S-adenosylmethionine
decarboxylase (ySAMdc) gene driven by a fruit-specific E8 promoter to investigate
the role of polyamines in fruit metabolism. Stable integration of E8-ySAMdc
chimeric gene in tomato genome led to ripening-specific accumulation of
polyamines, spermidine (Spd) and spermine (Spm), which in turn affected higher
accumulation of glutamine, asparagine, and organic acids in the red fruit with
significant decrease in the contents of valine, aspartate, sucrose, and glucose.
The metabolite profiling analysis suggests that Spd/Spm are perceived as
"signaling" organic-N metabolites by the fruit cells, resulting in the
stimulation of carbon sequestration; enhanced synthesis of biomolecules;
increased acid to sugar ratio, a good attribute for the fruit flavor; and in the
accumulation of another "vital amine," choline, which is an essential
micronutrient for brain development. A limited transcriptome analysis of the
transgenic fruit that accumulate higher polyamines revealed a large number of
differentially expressed genes, about 55% of which represented discrete
functional categories, and the remaining 45% were novel, unknown, or
unclassified: amino acid biosynthesis, carotenoid biosynthesis, cell wall
metabolism, chaperone family, flavonoid biosynthesis, fruit ripening, isoprenoid
biosynthesis, polyamine biosynthesis, signal transduction,
stress/defense-related, transcription, translation, and vacuolar function. There
was a good correspondence between some gene transcripts and their protein
products, but not in the case of the tonoplast intrinsic protein, which showed
post-transcriptional regulation. Higher metabolic activity of the transgenic
fruit is reflected in higher respiratory activity, and upregulation of chaperones
and mitochondrial cytochrome oxidase transcripts compared to the control. These
transgenic plants are a new resource to understand the role of Spd/Spm in fruit
biology. Transcriptome analysis and metabolic profiles of Spd/Spm accumulating,
transgenic fruit suggest the presence of an intricate regulation and
interconnection between certain metabolic pathways that are revived when Spd and
Spm likely reach a certain threshold. Thus, polyamines act as antiapoptotic
regulatory molecules and are able to revive metabolic memory in the tomato fruit.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17955994 [PubMed - indexed for MEDLINE]

235: J AOAC Int. 2007 Sep-Oct;90(5):1450-5.

Transgenic approaches for cyanogen reduction in cassava.

Siritunga D, Sayre R.

University of Puerto Rico Mayaguez, Department of Biology, Mayaguez, Puerto Rico.

For cassava to become a safe and acceptable crop, it is necessary to reduce the
cyanogen levels in cassava foods. While this objective can be achieved by
processing procedures, recent findings have shown that it is also possible to
achieve it by suppression of cyanogen synthesis or by accelerating cyanogen
turnover and volatilization. In 2003, cyanogen-free cultivars were generated by
selective inhibition CYP79D1/D2 gene expression. The CYP79D1/D2 enzymes catalyze
the first-dedicated step in cyanogen synthesis. Tissue-specific inhibition of
CYP79D1/D2 expression in leaves lead to a 99% reduction in root cyanogen levels,
indicating that the cyanogenic glycoside, linamarin, is synthesized in leaves and
transported to roots. An alternative strategy to the reduce cyanogen content is
to enhance cyanogen detoxification and cyanide volatilization during processing.
This strategy has the advantage that cyanogen levels in unprocessed roots are not
altered, potentially providing protection against herbivory andlor theft. To
produce cultivars that promote rapid cyanide volatilization, hydroxynitrile lyase
(HNL), which catalyzes the last step in cyanogenesis, was overexpressed in roots.
Elevated HNL activity resulted in a 3-fold increase in the rate of cyanogen
turnover. Importantly, the cyanogen content of the transformed and wild-type
plants was identical, a potential benefit for farmers.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17955993 [PubMed - indexed for MEDLINE]

236: J AOAC Int. 2007 Sep-Oct;90(5):1445-9.

Delivering golden rice to developing countries.

Mayer JE.

Golden Rice Project, Campus Technologies Freiburg, 79104 Freiburg, Germany.
jorge.mayer@goldenrice.org

Micronutrient deficiencies create a vicious circle of malnutrition, poverty, and
economic dependency that we must strive to break. Golden Rice offers a
sustainable solution to reduce the prevalence of vitamin A deficiency-related
diseases and mortality, a problem that affects the health of millions of children
in all developing countries. The technology is based on the reconstitution of the
carotenoid biosynthetic pathway by addition of 2 transgenes. The outcome of this
high-tech approach will be provided to end users as nutrient-dense rice varieties
that are agronomically identical to their own, locally adapted varieties. This
intervention has the potential to reach remote rural populations without access
to fortification and supplementation programs. As part of our delivery strategy,
we are partnering with government and nongovernment, national and international
agricultural institutions to navigate through cumbersome and expensive regulatory
regimes that affect the release of genetically modified crops, and to create
local demand for the biofortified rice varieties.

Publication Types:
Review

PMID: 17955992 [PubMed - indexed for MEDLINE]

237: J AOAC Int. 2007 Sep-Oct;90(5):1440-4.

Impact of foods nutritionally enhanced through biotechnology in alleviating
malnutrition in developing countries.

Gilani GS, Nasim A.

Nutrition Research Division, Food Directorate, Health Products and Food Branch,
Health Canada, Government of Canada, 251 Sir Frederick Banting Dwy, Ottawa, ON,
Canada. Sarwar_Gilani@hc-sc.gc.ca

According to United Nations (UN) projections, the world's population will grow
from 6.1 billion in 2000 to 8 billion in 2025 and 9.4 billion in 2050. Most (93%)
of the increase will take place in developing countries. The rapid population
growth in developing countries creates major challenges for governments regarding
food and nutrition security. According to current World Health Organization
estimates, more than 3 billion people worldwide, especially in developing
countries, are malnourished in essential nutrients. Malnutrition imposes severe
costs on a country's population due to impaired physical and cognitive abilities
and reduced ability to work. Little progress has been made in improving
malnutrition over the past few decades. The Food and Agriculture Organization of
the UN would like to see more nutrient-rich foods introduced into these
countries, because supplements are expensive and difficult to distribute widely.
Biofortification of staple crops through modern biotechnology can potentially
help in alleviating malnutrition in developing countries. Several genetically
modified crops, including rice, potatoes, oilseeds, and cassava, with elevated
levels of essential nutrients (such as vitamin A, iron, zinc, protein and
essential amino acids, and essential fatty acids); reduced levels of
antinutritional factors (such as cyanogens, phytates, and glycoalkaloid); and
increased levels of factors that influence bioavailability and utilization of
essential nutrients (such as cysteine residues) are advancing through field trial
stage and regulatory processes towards commercialization. The ready availability
and consumption of the biofortified crops would have a significant impact in
reducing malnutrition and the risk of chronic disease in developing countries.

Publication Types:
Review

PMID: 17955991 [PubMed - indexed for MEDLINE]

238: Plant J. 2008 Jan;53(1):42-52. Epub 2007 Oct 22.

Transcription factor AtTCP14 regulates embryonic growth potential during seed
germination in Arabidopsis thaliana.

Tatematsu K, Nakabayashi K, Kamiya Y, Nambara E.

Growth Regulation Research Group, RIKEN Plant Science Center, Yokohama, 1-7-22,
Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.

To understand the molecular mechanisms underlying regulation of seed germination,
we searched enriched cis elements in the upstream regions of Arabidopsis genes
whose transcript levels increased during seed germination. Using available
published microarray data, we found that two cis elements, Up1 or Up2, which
regulate outgrowth of Arabidopsis axillary shoots, were significantly
over-represented. Classification of Up1- and Up2-containing genes by gene
ontology revealed that protein synthesis-related genes, especially ribosomal
protein genes, were highly over-represented. Expression analysis using a reporter
gene driven by a synthetic promoter regulated by these elements showed that the
Up1 is necessary and sufficient for germination-associated gene induction,
whereas Up2 acts as an enhancer of Up1. Up1-mediated gene expression was
suppressed by treatments that blocked germination. Up1 is almost identical to the
site II motif, which is the predicted target of TCP transcription factors. Of 24
AtTCP genes, AtTCP14, which showed the highest transcript level just prior to
germination, was functionally characterized to test its involvement in the
regulation of seed germination. Transposon-tagged lines for AtTCP14 showed
delayed germination. In addition, germination of attcp14 mutants exhibited
hypersensitivity to exogenously applied abscisic acid and paclobutrazol, an
inhibitor of gibberellin biosynthesis. AtTCP14 was predominantly expressed in the
vascular tissues of the embryo, and affected gene expression in radicles in a
non-cell-autonomous manner. Taken together, these results indicate that AtTCP14
regulates the activation of embryonic growth potential in Arabidopsis seeds.

PMID: 17953649 [PubMed - indexed for MEDLINE]

239: PLoS Genet. 2007 Oct;3(10):1965-74.

Meiotic transmission of an in vitro-assembled autonomous maize minichromosome.

Carlson SR, Rudgers GW, Zieler H, Mach JM, Luo S, Grunden E, Krol C, Copenhaver
GP, Preuss D.

Chromatin, Chicago, Illinois, USA.

Autonomous chromosomes are generated in yeast (yeast artificial chromosomes) and
human fibrosarcoma cells (human artificial chromosomes) by introducing purified
DNA fragments that nucleate a kinetochore, replicate, and segregate to daughter
cells. These autonomous minichromosomes are convenient for manipulating and
delivering DNA segments containing multiple genes. In contrast, commercial
production of transgenic crops relies on methods that integrate one or a few
genes into host chromosomes; extensive screening to identify insertions with the
desired expression level, copy number, structure, and genomic location; and long
breeding programs to produce varieties that carry multiple transgenes. As a step
toward improving transgenic crop production, we report the development of
autonomous maize minichromosomes (MMCs). We constructed circular MMCs by
combining DsRed and nptII marker genes with 7-190 kb of genomic maize DNA
fragments containing satellites, retroelements, and/or other repeats commonly
found in centromeres and using particle bombardment to deliver these constructs
into embryogenic maize tissue. We selected transformed cells, regenerated plants,
and propagated their progeny for multiple generations in the absence of
selection. Fluorescent in situ hybridization and segregation analysis
demonstrated that autonomous MMCs can be mitotically and meiotically maintained.
The MMC described here showed meiotic segregation ratios approaching Mendelian
inheritance: 93% transmission as a disome (100% expected), 39% transmission as a
monosome crossed to wild type (50% expected), and 59% transmission in self
crosses (75% expected). The fluorescent DsRed reporter gene on the MMC was
expressed through four generations, and Southern blot analysis indicated the
encoded genes were intact. This novel approach for plant transformation can
facilitate crop biotechnology by (i) combining several trait genes on a single
DNA fragment, (ii) arranging genes in a defined sequence context for more
consistent gene expression, and (iii) providing an independent linkage group that
can be rapidly introgressed into various germplasms.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17953486 [PubMed - indexed for MEDLINE]

240: Genet Mol Res. 2007 Jun 30;6(2):445-52.

Gene flow from transgenic to nontransgenic soybean plants in the Cerrado region
of Brazil.

Abud S, de Souza PI, Vianna GR, Leonardecz E, Moreira CT, Faleiro FG, Jöºnior JN,
Monteiro PM, Rech EL, Aragö¸o FJ.

Embrapa Cerrados, Planaltina, DF, Brasil.

Evaluation of transgenic crops under field conditions is a fundamental step for
the production of genetically engineered varieties. In order to determine if
there is pollen dispersal from transgenic to nontransgenic soybean plants, a
field release experiment was conducted in the Cerrado region of Brazil.
Nontransgenic plants were cultivated in plots surrounding Roundup Ready
transgenic plants carrying the cp4 epsps gene, which confers herbicide tolerance
against glyphosate herbicide, and pollen dispersal was evaluated by checking for
the dominant gene. The percentage of cross-pollination was calculated as a
fraction of herbicide-tolerant and -nontolerant plants. The greatest amount of
transgenic pollen dispersion was observed in the first row, located at one meter
from the central (transgenic) plot, with a 0.52% average frequency. The frequency
of pollen dispersion decreased to 0.12% in row 2, reaching 0% when the plants
were up to 10 m distance from the central plot. Under these conditions pollen
flow was higher for a short distance. This fact suggests that the management
necessary to avoid cross-pollination from transgenic to nontransgenic plants in
the seed production fields should be similar to the procedures currently utilized
to produce commercial seeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17952868 [PubMed - indexed for MEDLINE]

241: Plant Cell. 2007 Oct;19(10):3100-10. Epub 2007 Oct 19.

MIDGET unravels functions of the Arabidopsis topoisomerase VI complex in DNA
endoreduplication, chromatin condensation, and transcriptional silencing.

Kirik V, Schrader A, Uhrig JF, Hulskamp M.

University of Cologne, Botanical Institute III, 50931 Cologne, Germany.
vkirik@stanford.edu

The plant homologs of the archaeal DNA topoisomerase VI complex are required for
the progression of endoreduplication cycles. Here, we describe the identification
of MIDGET (MID) as a novel component of topoisomerase VI. We show that mid
mutants show the same phenotype as rhl1, rhl2, and top6B mutants and that MID
protein physically interacts with RHL1. The phenotypic analysis revealed new
phenotypes, indicating that topoisomerase VI is involved in chromatin
organization and transcriptional silencing. In addition, genetic evidence is
provided suggesting that the ATR-dependent DNA damage repair checkpoint is
activated in mid mutants, and CYCB1;1 is ectopically activated. Finally, we
demonstrate that overexpression of CYCB1;2 can rescue the endoreduplication
defects in mid mutants, suggesting that in mid mutants, a specific checkpoint is
activated preventing further progression of endoreduplication cycles.

PMID: 17951446 [PubMed - indexed for MEDLINE]

242: Food Chem Toxicol. 2008 Jan;46(1):9-33. Epub 2007 Sep 14.

The application of post-market monitoring to novel foods.

Hepburn P, Howlett J, Boeing H, Cockburn A, Constable A, Davi A, de Jong N,
Moseley B, Oberdö¶rfer R, Robertson C, Wal JM, Samuels F.

Unilever, Safety and Environmental Assurance Centre, Colworth Park, Sharnbrook,
Befordshire MK44 1LQ, United Kingdom.

The role of post-market monitoring (PMM) in the safety assessment of novel foods
is critically discussed in order to derive guidelines as to in which situations
the application of PMM might be warranted. Available data sources on food
consumption and health status, and the methodologies for generating such data are
reviewed. The paper suggests improvements to make them more applicable for PMM
purposes. It is concluded that any PMM programme must be a hypothesis-driven
scientific exercise. PMM can have a role as a complement to, but not as a
replacement for, a comprehensive pre-market safety assessment. Its use may be
appropriate to confirm that product use is as predicted in the pre-market
assessment; to provide reassurance that effects observed in the pre-market
assessment occur with no greater frequency or intensity in the post-market phase
than anticipated; and to investigate the significance of any adverse effects
reported by consumers after market-launch. However PMM is insufficiently powerful
to test the hypothesis that any effects seen in the pre-market assessment are
absent in the post-market phase. Current methodologies place limitations on what
PMM can achieve. PMM should only be used when triggered by or when the focus is
on specific evidence-based questions.

Publication Types:
Review

PMID: 17950974 [PubMed - indexed for MEDLINE]

243: Genetics. 2007 Nov;177(3):1509-26. Epub 2007 Oct 18.

A transgenomic cytogenetic sorghum (Sorghum propinquum) bacterial artificial
chromosome fluorescence in situ hybridization map of maize (Zea mays L.)
pachytene chromosome 9, evidence for regions of genome hyperexpansion.

Amarillo FI, Bass HW.

Department of Biological Science, Florida State University, Tallahassee, Florida
32306-4370.

A cytogenetic FISH map of maize pachytene-stage chromosome 9 was produced with 32
maize marker-selected sorghum BACs as probes. The genetically mapped markers used
are distributed along the linkage maps at an average spacing of 5 cM. Each locus
was mapped by means of multicolor direct FISH with a fluorescently labeled probe
mix containing a whole-chromosome paint, a single sorghum BAC clone, and the
centromeric sequence, CentC. A maize-chromosome-addition line of oat was used for
bright unambiguous identification of the maize 9 fiber within pachytene
chromosome spreads. The locations of the sorghum BAC-FISH signals were
determined, and each new cytogenetic locus was assigned a centiMcClintock
position on the short (9S) or long (9L) arm. Nearly all of the markers appeared
in the same order on linkage and cytogenetic maps but at different relative
positions on the two. The CentC FISH signal was localized between cdo17 (at
9L.03) and tda66 (at 9S.03). Several regions of genome hyperexpansion on maize
chromosome 9 were found by comparative analysis of relative marker spacing in
maize and sorghum. This transgenomic cytogenetic FISH map creates anchors between
various maps of maize and sorghum and creates additional tools and information
for understanding the structure and evolution of the maize genome.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17947405 [PubMed - indexed for MEDLINE]

244: Appetite. 2008 Mar-May;50(2-3):340-52. Epub 2007 Sep 18.

Consumer responses to communication about food risk management.

van Dijk H, Houghton J, van Kleef E, van der Lans I, Rowe G, Frewer L.

Marketing and Consumer Behaviour Group, Department of Social Sciences, Wageningen
University, Hollandseweg 1, 6706 KN Wageningen, The Netherlands.
Heleen.vanDijk@wur.nl

Recent emphasis within policy circles has been on transparent communication with
consumers about food risk management decisions and practices. As a consequence,
it is important to develop best practice regarding communication with the public
about how food risks are managed. In the current study, the provision of
information about regulatory enforcement, proactive risk management, scientific
uncertainty and risk variability were manipulated in an experiment designed to
examine their impact on consumer perceptions of food risk management quality. In
order to compare consumer reactions across different cases, three food hazards
were selected (mycotoxins on organically grown food, pesticide residues, and a
genetically modified potato). Data were collected from representative samples of
consumers in Germany, Greece, Norway and the UK. Scores on the "perceived food
risk management quality" scale were subjected to a repeated-measures mixed linear
model. Analysis points to a number of important findings, including the existence
of cultural variation regarding the impact of risk communication
strategies-something which has obvious implications for pan-European risk
communication approaches. For example, while communication of uncertainty had a
positive impact in Germany, it had a negative impact in the UK and Norway.
Results also indicate that food risk managers should inform the public about
enforcement of safety laws when communicating scientific uncertainty associated
with risks. This has implications for the coordination of risk communication
strategies between risk assessment and risk management organizations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17945386 [PubMed - in process]

245: Crit Rev Food Sci Nutr. 2007;47(7):675-99.

A review on tomato authenticity: quality control methods in conjunction with
multivariate analysis (chemometrics).

Arvanitoyannis IS, Vaitsi OB.

University of Thessaly School of Agricultural Sciences Department of Agriculture
Animal Production & Aquatic Production, Volos, Hellas, Greece. parmenion@uth.gr

Authenticity and traceability have been two of the most important issues in the
food chain. Authenticity in particular, is closely related with both food quality
and safety issues. Vegetables stand for a category of foods heavily affected by
adulteration either in terms of geographic origin (national or international
level) or production methods (organic or conventional production, fertilizers,
pesticides, genetically modified vegetables). This review aims at addressing most
of the currently applied methods for ensuring quality control of vegetables; a)
instrumental: ion chromatography, high pressure liquid chromatography, atomic
absorption spectrophotometry, electronic nose and mass spectroscopy and b)
sensory analysis. The results of all the above mentioned methods were analyzed by
means of multivariate analysis (principal component analysis, discriminant
analysis, cluster analysis, canonical analysis, and factor analysis). All ensuing
results and conclusions are summarized in eight comprehensive tables.

Publication Types:
Review

PMID: 17943497 [PubMed - indexed for MEDLINE]

246: J Int Bioethique. 2006 Dec;17(4):109-14.

GMOs and development.

DaSilva EJ.

Section of Life Sciences Division of Basic and Engineering Sciences, UNESCO,
Paris, France.

PMID: 17939274 [PubMed - indexed for MEDLINE]

247: Nat Biotechnol. 2007 Nov;25(11):1277-9. Epub 2007 Oct 14.

Folate fortification of rice by metabolic engineering.

Storozhenko S, De Brouwer V, Volckaert M, Navarrete O, Blancquaert D, Zhang GF,
Lambert W, Van Der Straeten D.

Unit Plant Hormone Signalling and Bio-imaging, Department of Molecular Genetics,
Ghent University, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium.

Rice, the world's major staple crop, is a poor source of essential
micronutrients, including folates (vitamin B9). We report folate biofortification
of rice seeds achieved by overexpressing two Arabidopsis thaliana genes of the
pterin and para-aminobenzoate branches of the folate biosynthetic pathway from a
single locus. We obtained a maximal enhancement as high as 100 times above wild
type, with 100 g of polished raw grains containing up to four times the adult
daily folate requirement.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17934451 [PubMed - indexed for MEDLINE]

248: Appl Environ Microbiol. 2007 Dec;73(24):8012-7. Epub 2007 Oct 12.

Effect of feeding cows genetically modified maize on the bacterial community in
the bovine rumen.

Wiedemann S, Gö¼rtler P, Albrecht C.

Institute of Biochemistry and Molecular Medicine, University of Bern, Buehlstr.
28, CH-3012 Bern, Switzerland. christiane.albrecht@mci.unibe.ch

Rumen-cannulated cows (n = 4) were fed successively silage made from either
conventional or genetically modified (GM) maize. Results revealed no effects of
GM maize on the dynamics of six ruminal bacterial strains (investigated by
real-time PCR) compared to the conventional maize silage.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17933942 [PubMed - indexed for MEDLINE]

249: Plant Cell. 2007 Oct;19(10):3127-45. Epub 2007 Oct 12.

Subcellular localization and functional domain studies of DEFECTIVE KERNEL1 in
maize and Arabidopsis suggest a model for aleurone cell fate specification
involving CRINKLY4 and SUPERNUMERARY ALEURONE LAYER1.

Tian Q, Olsen L, Sun B, Lid SE, Brown RC, Lemmon BE, Fosnes K, Gruis DF,
Opsahl-Sorteberg HG, Otegui MS, Olsen OA.

Pioneer Hi-Bred International, A DuPont Business, Johnston, Iowa 50131, USA.

DEFECTIVE KERNEL1 (DEK1), which consists of a membrane-spanning region (DEK1-MEM)
and a calpain-like Cys proteinase region (DEK1-CALP), is essential for aleurone
cell formation at the surface of maize (Zea mays) endosperm. Immunolocalization
and FM4-64 dye incubation experiments showed that DEK1 and CRINKLY4 (CR4), a
receptor kinase implicated in aleurone cell fate specification, colocalized to
plasma membrane and endosomes. SUPERNUMERARY ALEURONE LAYER1 (SAL1), a negative
regulator of aleurone cell fate encoding a class E vacuolar sorting protein,
colocalized with DEK1 and CR4 in endosomes. Immunogold localization, dual-axis
electron tomography, and diffusion of fluorescent dye tracers showed that young
aleurone cells established symplastic subdomains through plasmodesmata of larger
dimensions than those connecting starchy endosperm cells and that CR4
preferentially associated with plasmodesmata between aleurone cells. Genetic
complementation experiments showed that DEK1-CALP failed to restore wild-type
phenotypes in maize and Arabidopsis thaliana dek1 mutants, and DEK1-MEM also
failed to restore wild-type phenotypes in Arabidopsis dek1-1 mutants. Instead,
ectopic expression of DEK1-MEM under the control of the cauliflower mosaic virus
35S promoter gave a dominant negative phenotype. These data suggest a model for
aleurone cell fate specification in which DEK1 perceives and/or transmits a
positional signal, CR4 promotes the lateral movement of aleurone signaling
molecules between aleurone cells, and SAL1 maintains the proper plasma membrane
concentration of DEK1 and CR4 proteins via endosome-mediated
recycling/degradation.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17933905 [PubMed - indexed for MEDLINE]

250: Plant Cell. 2007 Oct;19(10):3194-211. Epub 2007 Oct 12.

Erratum in:
Plant Cell. 2007 Dec;19(12):4131-2.

Manipulation of phytoene levels in tomato fruit: effects on isoprenoids,
plastids, and intermediary metabolism.

Fraser PD, Enfissi EM, Halket JM, Truesdale MR, Yu D, Gerrish C, Bramley PM.

School of Biological Sciences, Royal Holloway, University of London, Egham,
Surrey, TW20 OEX, United Kingdom.

In tomato (Solanum lycopersicum), phytoene synthase-1 (PSY-1) is the key
biosynthetic enzyme responsible for the synthesis of fruit carotenoids. To
further our understanding of carotenoid formation in tomato fruit, we
characterized the effect of constitutive expression of an additional tomato Psy-1
gene product. A quantitative data set defining levels of carotenoid/isoprenoid
gene expression, enzyme activities, and metabolites was generated from fruit that
showed the greatest perturbation in carotenoid content. Transcriptional
upregulation, resulting in increased enzyme activities and metabolites, occurred
only in the case of Psy-1, Psy-2, and lycopene cyclase B. For reactions involving
1-deoxy-d-xylulose5-phosphate synthase, geranylgeranyl diphosphate synthase,
phytoene desaturase, zeta-carotene desaturase, carotene isomerase, and lycopene
beta-cyclase, there were no correlations between gene expression, enzyme
activities, and metabolites. Perturbations in carotenoid composition were
associated with changes in plastid type and with chromoplast-like structures
arising prematurely during fruit development. The levels of >120 known
metabolites were determined. Comparison with the wild type illustrated that key
metabolites (sucrose, glucose/fructose, and Glu) and sectors of intermediary
metabolism (e.g., tricarboxylic [corrected] acid cycle intermediates and fatty
acids) in the Psy-1 transgenic mature green fruit resembled changes in metabolism
associated with fruit ripening. General fruit developmental and ripening
properties, such as ethylene production and fruit firmness, were unaffected.
Therefore, it appears that the changes to pigmentation, plastid type, and
metabolism associated with Psy-1 overexpression are not connected with the
ripening process.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17933904 [PubMed - indexed for MEDLINE]

251: Electrophoresis. 2007 Nov;28(22):4247-54.

Ultra-fast simultaneous analysis of genetically modified organisms in maize by
microchip electrophoresis with LIF detector.

Kumar KS, Kang SH.

Department of Chemistry and Research Institute Basic Science, Chonbuk National
University, Jeonju, South Korea.

This study examined the potential of microchip electrophoresis (ME) with a LIF
detector using a programmed field strength gradient (PFSG) in a conventional
glass double-T microchip for the ultra-fast detection and simultaneous analysis
of genetically modified (GM) maize. The separation efficiency and sensitivity at
various sieving gels (poly(ethylene oxide) (PEO, M(r) 8,000,000) and
2-hydroxyethylcellulose (HEC) (M(r) 250,000)) and fluorescent dye concentrations
were investigated. The PCR products of both the GM and non-GM maize were analyzed
within 30 s under the PFSG (470.6 V/cm for 20 s, 117.6 V/cm for 12 s, and 470.6
V/cm for 30 s) with a 2.5% HEC sieving matrix in the running buffer, 1 x
Tris-borate EDTA (TBE) (pH 8.30) and 0.5 ppm ethidium bromide. The five
transgenic maize varieties (Event176, MON810, Bt11, GA21, and T25) examined in
this study were also clearly differentiated by ME-PFSG within 30 s in a single
run without any loss of resolution. The ME-PFSG technique is a powerful tool for
the ultra-fast detection and simultaneous analysis of GMOs in a variety of foods
including maize.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17932874 [PubMed - indexed for MEDLINE]

252: Science. 2007 Oct 12;318(5848):265-8.

Lyso-phosphatidylcholine is a signal in the arbuscular mycorrhizal symbiosis.

Drissner D, Kunze G, Callewaert N, Gehrig P, Tamasloukht M, Boller T, Felix G,
Amrhein N, Bucher M.

Institute of Plant Sciences, Eidgenö¶ssische Technische Hochschule (ETH) Zurich,
Experimental Station Eschikon 33, 8315 Lindau, Switzerland.

The arbuscular mycorrhizal (AM) symbiosis represents the most widely distributed
mutualistic root symbiosis. We report that root extracts of mycorrhizal plants
contain a lipophilic signal capable of inducing the phosphate transporter genes
StPT3 and StPT4 of potato (Solanum tuberosum L.), genes that are specifically
induced in roots colonized by AM fungi. The same signal caused rapid
extracellular alkalinization in suspension-cultured tomato (Solanum lycopersicum
L.) cells and induction of the mycorrhiza-specific phosphate transporter gene
LePT4 in these cells. The active principle was characterized as the lysolipid
lyso-phosphatidylcholine (LPC) via a combination of gene expression studies,
alkalinization assays in cell cultures, and chromatographic and mass
spectrometric analyses. Our results highlight the importance of lysophospholipids
as signals in plants and in particular in the AM symbiosis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17932296 [PubMed - indexed for MEDLINE]

253: Dokl Biol Sci. 2007 Jul-Aug;415:267-9.

Expression of the artificial gene encoding anti-microbial peptide cecropin P1
increases the resistance of transgenic potato plants to potato blight and white
rot.

Zakharchenko NS, Rukavtsova EB, Gudkov AT, Yukhmanova AA, Shkol'naya LA, Kado CI,
Bur'yanov YI.

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Pushchino Branch),
Russian Academy of Sciences, Pushchino, Moscow oblast, 142290 Russia.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17929662 [PubMed - indexed for MEDLINE]

254: Plant Foods Hum Nutr. 2007 Dec;62(4):185-91. Epub 2007 Oct 11.

Towards generating caffeine-free tea by metabolic engineering.

Yadav SK, Ahuja PS.

Biotechnology Division, Institute of Himalayan Bioresource Technology, CSIR,
Palampur, H.P. 176061, India. skyt@rediffmail.com

Tea is a rich source of antioxidants which are contributing substantially to the
promotion of health and the prevention of various chronic diseases. Despite the
fact that tea has various important compounds, it also contains a purine
alkaloid, caffeine. High intake of tea leads to an increase in level of caffeine
in addition to its important antioxidant constituents. Increased level of
caffeine causes several health related problems. Therefore, tea can become a most
useful source of beneficial compounds, if only its caffeine level is either
decreased or eliminated all together from the plant itself. This could be
achieved through either of the techniques; overexpressing caffeine degradative
pathway genes or silencing caffeine biosynthesis pathway gene. The identification
and cloning of caffeine biosynthesis in tea and degradative genes in
microorganisms opens up the possibility of using genetic engineering to produce
naturally decaffeinated tea. Here we review these different strategies which can
be employed to make caffeine-free tea, a human health beneficial drink.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17929169 [PubMed - indexed for MEDLINE]

255: Biochem Biophys Res Commun. 2007 Nov 30;363(4):983-8. Epub 2007 Oct 2.

Tomato plants overexpressing CaKR1 enhanced tolerance to salt and oxidative
stress.

Seong ES, Cho HS, Choi D, Joung YH, Lim CK, Hur JH, Wang MH.

School of Biotechnology, Kangwon National University, Chuncheon, Kangwon-do
200-701, Republic of Korea.

CaKR1 from pepper leaves encodes an ankyrin repeat domain zinc finger that is
thought to be involved in transcriptional regulation in response to pathogens and
abiotic stresses. Transgenic tomato plants expressing CaKR1 show enhanced
resistance to Phytophthora infestans. In this study, we further characterized
this CaKR1-overexpressing transgenic tomato line. Morphologically, the leaves of
the transgenic plants were thicker than those of control plants. Overexpressed
transgenic plants also produced lower levels of free oxygen radicals, such as
superoxide (O2-) and hydrogen peroxide (H2O2), and showed enhanced resistance to
salinity and oxidative stress. In particular, transgenic plants produced higher
levels of transcripts encoding the pathogenesis-related (PR) proteins LePR1,
LePR2, and LePR3, as well as oxidative stress response proteins, such as
superoxide dismutase (LeSOD2) and ascorbate peroxidase (LeAPX2 and LeAPX3). These
results suggest that CaKR1 is a key signaling molecule regulating plant
antioxidant metabolism and defense responses.

PMID: 17927963 [PubMed - indexed for MEDLINE]

256: Plant Biotechnol J. 2007 Nov;5(6):759-67.

Maize streak virus-resistant transgenic maize: a first for Africa.

Shepherd DN, Mangwende T, Martin DP, Bezuidenhout M, Kloppers FJ, Carolissen CH,
Monjane AL, Rybicki EP, Thomson JA.

Department of Molecular and Cell Biology, University of Cape Town, Private Bag,
Rondebosch 7701, South Africa. dionne.shepherd@uct.ac.za

In this article, we report transgene-derived resistance in maize to the severe
pathogen maize streak virus (MSV). The mutated MSV replication-associated protein
gene that was used to transform maize showed stable expression to the fourth
generation. Transgenic T2 and T3 plants displayed a significant delay in symptom
development, a decrease in symptom severity and higher survival rates than
non-transgenic plants after MSV challenge, as did a transgenic hybrid made by
crossing T2 Hi-II with the widely grown, commercial, highly MSV-susceptible,
white maize genotype WM3. To the best of our knowledge, this is the first maize
to be developed with transgenic MSV resistance and the first all-African-produced
genetically modified crop plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17924935 [PubMed - indexed for MEDLINE]

257: Proc Natl Acad Sci U S A. 2007 Oct 9;104(41):16204-8. Epub 2007 Oct 8.

Comment in:
Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):E10; author reply E11. Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):E9; author reply E11.

Toxins in transgenic crop byproducts may affect headwater stream ecosystems.

Rosi-Marshall EJ, Tank JL, Royer TV, Whiles MR, Evans-White M, Chambers C,
Griffiths NA, Pokelsek J, Stephen ML.

Department of Biology, Loyola University Chicago, Chicago, IL 60626, USA.
erosi@luc.edu

Corn (Zea mays L.) that has been genetically engineered to produce the Cry1Ab
protein (Bt corn) is resistant to lepidopteran pests. Bt corn is widely planted
in the midwestern United States, often adjacent to headwater streams. We show
that corn byproducts, such as pollen and detritus, enter headwater streams and
are subject to storage, consumption, and transport to downstream water bodies.
Laboratory feeding trials showed that consumption of Bt corn byproducts reduced
growth and increased mortality of nontarget stream insects. Stream insects are
important prey for aquatic and riparian predators, and widespread planting of Bt
crops has unexpected ecosystem-scale consequences.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17923672 [PubMed - indexed for MEDLINE]

258: Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16450-5. Epub 2007 Oct 8.

Plant nuclear factor Y (NF-Y) B subunits confer drought tolerance and lead to
improved corn yields on water-limited acres.

Nelson DE, Repetti PP, Adams TR, Creelman RA, Wu J, Warner DC, Anstrom DC, Bensen
RJ, Castiglioni PP, Donnarummo MG, Hinchey BS, Kumimoto RW, Maszle DR, Canales
RD, Krolikowski KA, Dotson SB, Gutterson N, Ratcliffe OJ, Heard JE.

Monsanto Company, 62 Maritime Drive, Mystic, CT 06355, USA.
donald.e.nelson@monsanto.com

Commercially improved crop performance under drought conditions has been
challenging because of the complexity of the trait and the multitude of factors
that influence yield. Here we report the results of a functional genomics
approach that identified a transcription factor from the nuclear factor Y (NF-Y)
family, AtNF-YB1, which acts through a previously undescribed mechanism to confer
improved performance in Arabidopsis under drought conditions. An orthologous
maize transcription factor, ZmNF-YB2, is shown to have an equivalent activity.
Under water-limited conditions, transgenic maize plants with increased ZmNF-YB2
expression show tolerance to drought based on the responses of a number of
stress-related parameters, including chlorophyll content, stomatal conductance,
leaf temperature, reduced wilting, and maintenance of photosynthesis. These
stress adaptations contribute to a grain yield advantage to maize under
water-limited environments. The application of this technology has the potential
to significantly impact maize production systems that experience drought.

PMID: 17923671 [PubMed - indexed for MEDLINE]

259: Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16402-9. Epub 2007 Oct 8.

Strategies for developing Green Super Rice.

Zhang Q.

National Key Laboratory of Crop Genetic Improvement, National Center of Plant
Gene Research and National Center of Crop Molecular Breeding, Huazhong
Agricultural University, Wuhan 430070, China. qifazh@mail.hzau.edu.cn

From a global viewpoint, a number of challenges need to be met for sustainable
rice production: (i) increasingly severe occurrence of insects and diseases and
indiscriminate pesticide applications; (ii) high pressure for yield increase and
overuse of fertilizers; (iii) water shortage and increasingly frequent occurrence
of drought; and (iv) extensive cultivation in marginal lands. A combination of
approaches based on the recent advances in genomic research has been formulated
to address these challenges, with the long-term goal to develop rice cultivars
referred to as Green Super Rice. On the premise of continued yield increase and
quality improvement, Green Super Rice should possess resistances to multiple
insects and diseases, high nutrient efficiency, and drought resistance, promising
to greatly reduce the consumption of pesticides, chemical fertilizers, and water.
Large efforts have been focused on identifying germplasms and discovering genes
for resistance to diseases and insects, N- and P-use efficiency, drought
resistance, grain quality, and yield. The approaches adopted include screening of
germplasm collections and mutant libraries, gene discovery and identification,
microarray analysis of differentially regulated genes under stressed conditions,
and functional test of candidate genes by transgenic analysis. Genes for almost
all of the traits have now been isolated in a global perspective and are
gradually incorporated into genetic backgrounds of elite cultivars by molecular
marker-assisted selection or transformation. It is anticipated that such
strategies and efforts would eventually lead to the development of Green Super
Rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17923667 [PubMed - indexed for MEDLINE]

260: BMC Bioinformatics. 2007 Oct 9;8:375.

Computational analysis of the relationship between allergenicity and
digestibility of allergenic proteins in simulated gastric fluid.

Jiang B, Qu H, Hu Y, Ni T, Lin Z.

College of Life Sciences, National Lab of Protein Engineering and Genetic
Engineering of Plants, Peking University, Beijing 100871, PR China.
jiangbingjun@gmail.com

BACKGROUND: Safety assessment of genetically modified (GM) food, with regard to
allergenic potential of transgene-encoded xenoproteins, typically involves
several different methods, evaluation by digestibility being one thereof.
However, there are still debates about whether the allergenicity of food
allergens is related to their resistance to digestion by the gastric fluid. The
disagreements may in part stem from classification of allergens only by their
sources, which we believe is inadequate, and the difficulties in achieving
identical experimental conditions for studying digestion by simulated gastric
fluid (SGF) so that results can be compared. Here, we reclassify allergenic food
allergens into alimentary canal-sensitized (ACS) and non-alimentary
canal-sensitized (NACS) allergens and use a computational model that simulates
gastric fluid digestion to analyze the digestibilities of these two types.
RESULTS: The model presented in this paper is as effective as SGF digestion
experiments, but more stable and reproducible. On the basis of this model, food
allergens are satisfactorily classified as ACS and NACS types by their pathways
for sensitization; the former are relatively resistant to gastric fluid digestion
while the later are relatively labile. CONCLUSION: The results suggest that it is
better to classify allergens into ACS and NACS types when understanding the
relationship between their digestibility and allergenicity and the digestibility
of a target foreign protein is a parameter for evaluating its allergenicity
during safety assessments of GM food.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17922925 [PubMed - indexed for MEDLINE]

261: FEBS J. 2007 Nov;274(21):5659-68. Epub 2007 Oct 8.

The hydroxyproline motif of male sex peptide elicits the innate immune response
in Drosophila females.

Domanitskaya EV, Liu H, Chen S, Kubli E.

Zoologisches Institut Universitö¤t Zö¼rich-Irchel, Zö¼rich, Switzerland.

Seminal fluid elicits a variety of physiological and behavioral changes in insect
females. In Drosophila melanogaster females, sex peptide (SP) is the major
seminal agent eliciting oviposition and reduction of receptivity. But SP also has
many other effects; for example, it stimulates food intake, egg production,
ovulation, juvenile hormone production and antimicrobial peptide synthesis. Thus,
SP very probably has several receptors. To identify putative targets and
signaling cascades, we studied the genome-wide regulation of genes by microarray
analysis of RNA isolated from females after mating with wild-type males or males
lacking SP, respectively. In addition, we studied the effects of SP on the
proteome of females. Sex peptide regulates gene activity differentially in the
head and in the abdomen. Genes coding for unspecific antimicrobial peptides are
specifically transcribed in the abdomen, e.g. the antimicrobial peptide drosocin
in epithelial tissues of the female genital tract (oviduct and calyx). Hence, SP
elicits a systemic [Peng J, Zipperlen P & Kubli E (2005) Curr Biol15, 1690-1694]
and an epithelial immune response. Ectopic expression of SP in the fat body of
transgenic virgin females (with subsequent secretion into the hemolymph) does not
elicit drosocin synthesis in the genital tract. Thus, the receptors for the
stimulation of the systemic and the epithelial responses by SP are
compartmentalized. The hydroxyproline (P*) motif of SP, P*TKFP*IP*SP*NP*, is
identified as a novel elicitor of the innate immune response. We suggest that SP
acts by chemical mimicry of sugar components of the bacterial cell wall. Thus, SP
may induce the immune system via pattern recognition receptors.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17922838 [PubMed - indexed for MEDLINE]

262: Vet Res Commun. 2007 Aug;31 Suppl 1:385-8.

Detection of genetically modified organisms in food: comparison among three
different DNA extraction methods.

Vodret B, Milia M, Orani MG, Serratrice G, Mancuso MR.

Zooprofilattic Institute of Sardinia, Feed Hygiene Unit, Sassari, Italy.
bruna.vodret@izs-sardegna.it

PMID: 17682920 [PubMed - indexed for MEDLINE]

263: Nat Biotechnol. 2007 Oct;25(10):1065-6.

Erratum in:
Nat Biotechnol. 2008 Feb;26(2):241.

Europe's anti-GM stance to presage animal feed shortage?

Mitchell P.

Publication Types:
News

PMID: 17921975 [PubMed - indexed for MEDLINE]

264: Plant Physiol. 2007 Dec;145(4):1408-22. Epub 2007 Oct 5.

Silencing of the mitochondrial ascorbate synthesizing enzyme
L-galactono-1,4-lactone dehydrogenase affects plant and fruit development in
tomato.

Alhagdow M, Mounet F, Gilbert L, Nunes-Nesi A, Garcia V, Just D, Petit J,
Beauvoit B, Fernie AR, Rothan C, Baldet P.

Institut National de la Recherche Agronomique, Universitö© Bordeaux 1, Universitö©
Victor Sö©galen-Bordeaux 2, BP 81, 33883 Villenave d'Ornon cedex, France.

L-Galactono-1,4-lactone dehydrogenase (EC 1.3.2.3) catalyzes the last step in the
main pathway of vitamin C (L-ascorbic acid) biosynthesis in higher plants. In
this study, we first characterized the spatial and temporal expression of
SlGalLDH in several organs of tomato (Solanum lycopersicum) plants in parallel
with the ascorbate content. P(35S):Slgalldh(RNAi) silenced transgenic tomato
lines were then generated using an RNAi strategy to evaluate the effect of any
resulting modification of the ascorbate pool on plant and fruit development. In
all P(35S):Slgalldh(RNAi) plants with reduced SlGalLDH transcript and activity,
plant growth rate was decreased. Plants displaying the most severe effects (dwarf
plants with no fruit) were excluded from further analysis. The most affected
lines studied exhibited up to an 80% reduction in SlGalLDH activity and showed a
strong reduction in leaf and fruit size, mainly as a consequence of reduced cell
expansion. This was accompanied by significant changes in mitochondrial function
and altered ascorbate redox state despite the fact that the total ascorbate
content remained unchanged. By using a combination of transcriptomic and
metabolomic approaches, we further demonstrated that several primary, like the
tricarboxylic acid cycle, as well as secondary metabolic pathways related to
stress response were modified in leaves and fruit of P(35S):Slgalldh(RNAi)
plants. When taken together, this work confirms the complexity of ascorbate
regulation and its link with plant metabolism. Moreover, it strongly suggests
that, in addition to ascorbate synthesis, GalLDH could play an important role in
the regulation of cell growth-related processes in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17921340 [PubMed - indexed for MEDLINE]

265: Curr Biol. 2007 Oct 23;17(20):1809-16. Epub 2007 Oct 4.

Sugar receptors in Drosophila.

Slone J, Daniels J, Amrein H.

Duke University Medical Center, Department of Molecular Genetics and
Microbiology, Durham, North Carolina 27710, USA.

The detection and discrimination of chemical compounds in potential foods are
essential sensory processes when animals feed. The fruit fly Drosophila
melanogaster employs 68 different gustatory receptors (GRs) for the detection of
mostly nonvolatile chemicals that include sugars, a diverse group of toxic
compounds present in many inedible plants and spoiled foods, and pheromones
[1-6]. With the exception of a trehalose (GR5a) and a caffeine (GR66a) receptor
[7-9], the functions of GRs involved in feeding are unknown. Here, we show that
the Gr64 genes encode receptors for numerous sugars. We generated a fly strain
that contained a deletion for all six Gr64 genes (DeltaGr64) and showed that
these flies exhibit no or a significantly diminished proboscis extension reflex
(PER) response when stimulated with glucose, maltose, sucrose, and several other
sugars. The only considerable response was detected when Gr64 mutant flies were
stimulated with fructose. Interestingly, response to trehalose is also abolished
in these flies, even though they contain a functional Gr5a gene, which has been
previously shown to encode a receptor for this sugar [8, 9]. This observation
indicates that two or more Gr genes are necessary for trehalose detection,
suggesting that GRs function as multimeric receptor complexes. Finally, we
present evidence that some members of the Gr64 gene family are transcribed as a
polycistronic mRNA, providing a mechanism for the coexpression of multiple sugar
receptors in the same taste neurons.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17919910 [PubMed - indexed for MEDLINE]

266: Biol Pharm Bull. 2007 Oct;30(10):1913-7.

The glycosylation and pharmacokinetics of CTLA4Ig produced in rice cells.

Kim HJ, Kim HJ.

College of Pharmacy, Chung-Ang University, Seoul, South Korea.

Cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4Ig) has immunosuppressive
activity and the ability to induce immune tolerance. There has been no report of
its glycosylation ratio or of the role of its glycans. We investigated the
terminal sialylation of rice cell-derived recombinant human CTLA4Ig (rrhCTLA4Ig)
using lectins. The glycosylation ratios of rrhCTLA4Ig and Chinese hamster ovary
(CHO) cell-derived recombinant human CTLA4Ig (crhCTLA4Ig) were evaluated by
chemical deglycosylation. After intravenous (i.v.) or subcutaneous (s.c.)
administration to rats, the pharmacokinetics of rrhCTLA4Ig and crhCTLA4Ig as well
as of their deglycosylated forms were evaluated. rrhCTLA4Ig does not have
terminal sialic acids and its glycosylation ratio was slightly lower than that of
crhCTLA4Ig. Its terminal elimination half-life (T(1/2)) was shorter than that of
crhCTLA4Ig following i.v. administration. However, the half-life was
significantly prolonged and was similar with that of crhCTLA4Ig following s.c.
administration. Moreover, the deglycosylated forms of both preparations were
cleared from the circulation faster than the native forms. These results suggest
that the presence of glycans on rrhCTLA4Ig and crhCTA4Ig are important for their
in vivo stability. In addition, the glycan structure of rrhCTLA4Ig is more
effective in maintaining in vivo stability after s.c. administration than after
i.v. administration although the glycans on rrhCTLA4Ig lack terminal sialic
acids, suggesting that its glycans have the potential for in vivo stability.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17917261 [PubMed - indexed for MEDLINE]

267: J Exp Bot. 2007;58(12):3213-26. Epub 2007 Oct 4.

Decreased shoot stature and grain alpha-amylase activity following ectopic
expression of a gibberellin 2-oxidase gene in transgenic wheat.

Appleford NE, Wilkinson MD, Ma Q, Evans DJ, Stone MC, Pearce SP, Powers SJ,
Thomas SG, Jones HD, Phillips AL, Hedden P, Lenton JR.

Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom.

Ectopic expression of a gibberellin 2-oxidase gene (PcGA2ox1) decreased the
content of bioactive gibberellins (GAs) in transgenic wheat, producing a range of
dwarf plants with different degrees of severity. In at least one case, a single
transformation event gave rise to T(1) plants with different degrees of dwarfism,
the phenotypes being stably inherited over at least four generations. The dwarf
phenotype, which included dark-green leaves, increased tillering and, in severe
cases, a prostrate growth habit, was replicated by the application of a GA
biosynthesis inhibitor to the wild type. Ear rachis length, grain set, and grain
size were also decreased in the wheat transformants, compared with an azygous
(null) line. The extent of post-germination alpha-amylase production in grains
reflected the severity of the shoot phenotype of the transformants and both
developmental processes were restored to normal by the application of gibberellic
acid (GA(3)). Expression of two GA biosynthesis genes (TaGA20ox1 and TaGA3ox2)
was up-regulated, and that of two alpha-amylase gene families (alpha-Amy1 and
alpha-Amy2) down regulated, in scutella of semi-dwarf lines, compared with
controls. The marked decline in transcript abundance of both alpha-amylase gene
families in aleurone was associated with a decreased content of bioactive GAs in
grains of the semi-dwarf lines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17916639 [PubMed - indexed for MEDLINE]

268: Bull Entomol Res. 2007 Oct;97(5):437-44.

F2 screen for resistance to a Bacillus thuringiensis-maize hybrid in the
sugarcane borer (Lepidoptera: Crambidae).

Huang FN, Leonard BR, Andow DA.

Department of Entomology, 404 Life Sciences Building, Louisiana State University
AgCenter, Baton Rouge, LA 70803, USA. fhuang@agcenter.lsu.edu

A novel F2 screening technique was developed for detecting resistance in
sugarcane borer, Diatraea saccharalis (F.), to transgenic Bacillus thuringiensis
(Bt)-maize expressing the Cry1Ab insecticidal protein. The F2 screening method
involved (i) collecting larvae from maize fields; (ii) establishing two-parent
families; (iii) screening F2 neonates for survival on Bt-maize leaf tissues; and
(iv) confirming resistance on commercial Bt-maize plants. With the F2 screening
method, 213 iso-line families of D. saccharalis were established from field
collections in northeast Louisiana, USA and were screened for Bt resistance. One
family was confirmed to carry a major Bt resistance allele(s). In a laboratory
bioassay, larval mortality of the Bt-resistant D. saccharalis on Bt-maize leaf
tissues was significantly lower than that of a Bt-susceptible strain. This
Bt-resistant D. saccharalis population is the first corn stalk borer species that
has completed larval development on commercial Bt-maize. The F2 screening
protocol developed in this study could be modified for detecting Bt resistance
alleles in other similar corn stalk borers, such as the European corn borer,
Ostrinia nubilalis (Hö¼bner), and the southwestern corn borer, D. grandiosella
Dyar.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17916262 [PubMed - indexed for MEDLINE]

269: Plant J. 2007 Dec;52(6):1080-93. Epub 2007 Oct 3.

Potassium transport systems in the moss Physcomitrella patens: pphak1 plants
reveal the complexity of potassium uptake.

Garciadeblas B, Barrero-Gil J, Benito B, Rodrö­guez-Navarro A.

Departamento de Biotecnologö­a, Universidad Politö©cnica de Madrid, 28040 Madrid,
Spain.

Potassium uptake is one of the most basic processes of plant physiology. However,
a comprehensive description is lacking. At a cellular level fungi have provided a
helpful but imperfect plant model, which we aim to improve using Physcomitrella
patens. Blast searches in expressed sequence tag databases demonstrated that
Physcomitrella expresses the same families of K(+) and Na(+) transport systems as
flowering plants. We cloned two inward rectifier channels, PpAKT1-2, and four
HAK-type transporters (PpHAK1-4). In both types of transport system, phylogenetic
analyses revealed that despite their high sequence conservation they could not be
included in Arabidopsis or rice (Oryza sativa) clusters. Both inward rectifier
channels and one HAK transporter (PpHAK1) were expressed in yeast. PpAKT1 and
activated mutants of PpAKT2 and PpHAK1 showed clear functions that were similar
to those of homologous systems of flowering plants. A pphak1 null mutant line of
Physcomitrella failed to deplete K(+) below 10 mum. Moreover, in a
non-K(+)-limiting medium in which wild-type plants grew only as protonema,
pphak1-1 plants produced leafy gametophores and contained 60% more K(+). We found
that Physcomitrella takes up K(+) through several systems. PpHAK1 is the dominant
system in plants that underwent K(+) starvation for long periods but an as-yet
unidentified system, which is non-selective for K(+), Rb(+), and Cs(+), dominates
in many other conditions. Finally, we discuss that, similar to PpHAK1, one of the
functions of AtHAK5 may be to control cellular K(+) content and that a
non-selective as-yet unidentified system also exists in Arabidopsis.

PMID: 17916113 [PubMed - indexed for MEDLINE]

270: Plant J. 2007 Dec;52(6):1027-40. Epub 2007 Oct 3.

Absence of the endo-beta-1,4-glucanases Cel1 and Cel2 reduces susceptibility to
Botrytis cinerea in tomato.

Flors V, Leyva Mde L, Vicedo B, Finiti I, Real MD, Garcö­a-Agustö­n P, Bennett AB,
Gonzö¡lez-Bosch C.

Area de Fisiologö­a Vegetal, Departamento de Ciencias Agrarias y del Medio
Natural, ESTCE, Universitat Jaume I, 12071 Castellö¨n, Spain.

Cel1 and Cel2 are members of the tomato (Solanum lycopersicum Mill)
endo-beta-1,4-glucanase (EGase) family that may play a role in fruit ripening and
organ abscission. This work demonstrates that Cel1 protein is present in other
vegetative tissues and accumulates during leaf development. We recently reported
the downregulation of both the Cel1 mRNA and protein upon fungal infection,
suggesting the involvement of EGases in plant-pathogen interactions. This
hypothesis was confirmed by assessing the resistance to Botrytis cinerea
infection of transgenic plants expressing both genes in an antisense orientation
(Anti-Cel1, Anti-Cel2 and Anti-Cel1-Cel2). The Anti-Cel1-Cel2 plants showed
enhanced resistance to this fungal necrotroph. Microscopical analysis of infected
leaves revealed that tomato plants accumulated pathogen-inducible callose within
the expanding lesion. Anti-Cel1-Cel2 plants presented a faster and enhanced
callose accumulation against B. cinerea than wild-type plants. The inhibitor
2-deoxy-d-glucose, a callose synthesis inhibitor, showed a direct relationship
between faster callose accumulation and enhanced resistance to B. cinerea. EGase
activity appears to negatively modulate callose deposition. The absence of both
EGase genes was associated with changes in the expression of the pathogen-related
genes PR1 and LoxD. Interestingly, Anti-Cel1-Cel2 plants were more susceptible to
Pseudomonas syringae, displaying severe disease symptoms and enhanced bacterial
growth relative to wild-type plants. Analysis of the involvement of Cel1 and Cel2
in the susceptibility to B. cinerea in fruits was done with the ripening-impaired
mutants Never ripe (Nr) and Ripening inhibitor (rin). The data reported in this
work support the idea that enzymes involved in cell wall metabolism play a role
in susceptibility to pathogens.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17916112 [PubMed - indexed for MEDLINE]

271: Mol Biotechnol. 2007 Jun;36(2):102-12.

Cloning, characterization, and transformation of the phosphoethanolamine
N-methyltransferase gene (ZmPEAMT1) in maize (Zea mays L.).

Wu S, Yu Z, Wang F, Li W, Ye C, Li J, Tang J, Ding J, Zhao J, Wang B.

The State Key Laboratory of Plant Genomics, Institute of Genetics and
Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China.

N-methylation of phosphoethanolamine, the committing step in choline (Cho)
biosynthesis in plants, is catalyzed by S-adenosyl-L-methionine:
phosphoethanolamine N-methyltransferase (PEAMT, EC 2.1.1.103). Herein we report
the cloning and characterization of the novel maize phosphoethanolamine
N-methyltransferase gene (ZmPEAMT1) using a combination of bioinformatics and a
PCR-based allele mining strategy. The cDNA sequence of ZmPEAMT1 gene is 1,806 bp
in length and translates a 495 amino acids peptide. The upstream promoter
sequence of ZmPEAMT1 were obtained by TAIL-PCR, and contained four kinds of
putative cis-acting regulatory elements, including stress-responsive elements,
phytohormone-responsive elements, pollen developmental special activation
elements, and light-induced signal transduction elements, as well as several
other structural features in common with the promoter of rice and Arabidopsis
homologies. RT-PCR analysis showed that expression of ZmPEAMT1 was induced by
salt stress and suppressed by high temperature. Over-expression of ZmPEAMT1
enhanced the salt tolerance, root length, and silique number in transgenic
Arabidopsis. These data indicated that ZmPEAMT1 maybe involved in maize root
development and stress resistance, and maybe having a potential application in
maize genetic engineering.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17914189 [PubMed - indexed for MEDLINE]

272: Z Naturforsch [C]. 2007 Jul-Aug;62(7-8):583-91.

Transgenic rice plants expressing a novel antifreeze glycopeptide possess
resistance to cold and disease.

Zhang S, Wei Y, Pan H.

College of Plant Science, Jilin University, Changchun/Jilin 130062, China.
zhang_sh@jlu.edu.cn

Freezing injury and disease are both restrictive factors in crop production. In
order to improve the tolerance ability to these stresses, a better way is to
carry out genetic engineering by transferring dualfunctional genes. A predicted
rice antifreeze glycopeptide gene was purposefully selected from rice
blast-induced cDNA library. Northern blot demonstrated that the gene is expressed
not only in blast-infected rice leaves, but also in low temperature-treated rice.
In addition, the expressed protein in Escherichia coli exhibits strong antifreeze
activities. The gene was overexpressed in rice plants transformed via
Agrobacterium tumefacient EHA105. Overall 112 T0 transformants were obtained in
this research. Cold tolerance and disease resistance of T1 transformants were,
respectively, investigated. The results showed that plants containing
overexpressed transgene can withstand -1 degrees C for 24 h without severe
chilling injury after thawed, and that disease symptoms of the parallel
transformants are highly reduced in response to blast infection, when compared
with controls. The relationship of the gene and several pathogenesis-related
protein genes to be chosen was analyzed and discussed. All these results
confirmed the dual role of the cloned gene, and implied that genetic engineering
using this kind of gene is a promising method to reduce biotic and abiotic
stresses.

PMID: 17913077 [PubMed - indexed for MEDLINE]

273: J Plant Physiol. 2008 Jan;165(1):71-82. Epub 2007 Oct 1.

Efficient generation of transgenic barley: the way forward to modulate
plant-microbe interactions.

Hensel G, Valkov V, Middlefell-Williams J, Kumlehn J.

Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Plant
Reproductive Biology, Corrensstr. 3, 06466 Gatersleben, Germany.
hensel@ipk.gatersleben.de

Stable genetic transformation represents the gold standard approach to the
detailed elucidation of plant gene functions. This is particularly relevant in
barley, an important experimental model widely employed in applied molecular,
genetic and cell biological research, and biotechnology. Presented are details of
the establishment of a protocol for Agrobacterium-mediated gene transfer to
immature embryos, which enables the highly efficient generation of transgenic
barley. Advancements were achieved through comparative experiments on the
influence of various explant treatments and co-cultivation conditions. The
analysis of representative numbers of transgenic lines revealed that the obtained
T-DNA copy numbers are typically low, the generative transmission of the
recombinant DNA is in accordance with the Mendelian rules and the vast majority
of the primary transgenics produce progeny that expresses the respective
transgene product. Moreover, the newly established protocol turned out to be
useful to transform not only the highly amenable cultivar (cv.) 'Golden Promise'
but also other spring and winter barley genotypes, albeit with substantially
lower efficiency. As a major result of this study, a very useful tool is now
available for future functional gene analyses as well as genetic engineering
approaches. With the aim to modify the expression of barley genes putatively
involved in plant-fungus interactions, numerous transgenic plants have been
generated using diverse expression cassettes. These plants represent an example
of how transformation technology may contribute to further our understanding of
important biological processes.

PMID: 17905476 [PubMed - indexed for MEDLINE]

274: Anal Biochem. 2007 Dec 15;371(2):194-200. Epub 2007 Aug 28.

A multiway approach to analyze metabonomic data: a study of maize seeds
development.

Castro C, Manetti C.

Dipartimento di Chimica, Universitö  degli Studi di RomaLa Sapienza, Piazzale Aldo
Moro, 5, 00185 Rome, Italy.

The aim of this research was to show that the application of multiway partial
least square-discriminant analysis to nuclear magnetic resonance spectra is a
valuable tool to analyze metabonomic data of transgenic maize. We evaluated the
effects, on the development of seeds, of the introduction of the
antisense-mediated downregulation and overexpression of the Rpd3 gene (ZmRpd3) in
the genome of a maize inbred line, we identified the metabolites involved in the
differentiation between classes of samples, directly integrating the evolution of
each metabolic perturbation over time in the model. Major differences were found
at the beginning of development, confirming the results obtained by transcript
analysis: ZmRpd3 transcripts and proteins accumulate during the initial stage of
development, suggesting a role for this gene in cell cycle control.

Publication Types:
Evaluation Studies

PMID: 17904514 [PubMed - indexed for MEDLINE]

275: Ann Bot (Lond). 2008 Jan;101(2):301-10. Epub 2007 Sep 27.

Ethylene and the regulation of senescence processes in transgenic Nicotiana
sylvestris plants.

Yang TF, Gonzalez-Carranza ZH, Maunders MJ, Roberts JA.

Division of Plant Sciences, School of Biosciences, University of Nottingham,
Sutton Bonington Campus, Loughborough, Leicestershire. LE12 5RD.

BACKGROUND AND AIMS: Exposure of plants to ethylene can influence a spectrum of
developmental processes including organ senescence and abscission. The aim of
this study was to examine the role of the gaseous regulator in Nicotiana
sylvestris plants exhibiting a silenced or constitutive ethylene response.
METHODS: Transgenic N. sylvestris plants were generated that either ectopically
expressed the Arabidopsis mutant ethylene receptor ETR1-1 or the tomato EIN3-like
(LeEIL1) gene. Highly expressing homozygous lines were selected and the
time-course of development, from germination to organ senescence, was studied.
KEY RESULTS: Fifty percent of the homozygous Pro(35S):ETR1-1 lines examined
showed a high susceptibility to collapse prior to flowering, with plant death
occurring within a few days of leaf wilting. The time-course of leaf senescence
in the remaining Pro(35S):ETR1-1 lines was visibly arrested compared to wild type
(negative segregant) plants and this observation was reaffirmed by chlorophyll
and protein analysis. Petal necrosis was also delayed in Pro(35S):ETR1-1 lines
and corolla abscission did not take place. When senescence of Pro(35S):ETR1-1
plants did take place this was accompanied by leaf bleaching, but tissues
remained fully turgid and showed no signs of collapse. A single Pro(35S):LeEIL1
line was found to exhibit consistently accelerated leaf and flower senescence and
precocious flower bud shedding. CONCLUSIONS: These observations support a role
for ethylene in regulating a spectrum of developmental events associated with
organ senescence and tissue necrosis. Furthermore, the transgenic lines generated
during this study may provide a valuable resource for exploring how senescence
processes are regulated in plants.

PMID: 17901061 [PubMed - indexed for MEDLINE]

276: Food Chem Toxicol. 2007 Nov;45(11):2073-85. Epub 2007 Aug 30.

Report of an Expert Panel on the reanalysis by of a 90-day study conducted by
Monsanto in support of the safety of a genetically modified corn variety (MON
863).

Doull J, Gaylor D, Greim HA, Lovell DP, Lynch B, Munro IC.

Pharmacology, Toxicology and Therapeutics, Division of Toxicology, Department of
Pharmacology, The University of Kansas Medical Center, 1018A Briedenthal
Building, 3901 Rainbow Boulevard, Kansas City, KS 66160-7417, USA.

MON 863, a genetically engineered corn variety that contains the gene for
modified Bacillus thuringiensis Cry3Bb1 protein to protect against corn rootworm,
was tested in a 90-day toxicity study as part of the process to gain regulatory
approval. This study was reanalyzed by Sö©ralini et al. who contended that the
study showed possible hepatorenal effects of MON 863. An Expert Panel was
convened to assess the original study results as analyzed by the Monsanto Company
and the reanalysis conducted by Sö©ralini et al. The Expert Panel concludes that
the Sö©ralini et al. reanalysis provided no evidence to indicate that MON 863 was
associated with adverse effects in the 90-day rat study. In each case,
statistical findings reported by both Monsanto and Sö©ralini et al. were
considered to be unrelated to treatment or of no biological or clinical
importance because they failed to demonstrate a dose-response relationship,
reproducibility over time, association with other relevant changes (e.g.,
histopathology), occurrence in both sexes, difference outside the normal range of
variation, or biological plausibility with respect to cause-and-effect. The
Sö©ralini et al. reanalysis does not advance any new scientific data to indicate
that MON 863 caused adverse effects in the 90-day rat study.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17900781 [PubMed - indexed for MEDLINE]

277: Int J Food Microbiol. 2007 Oct 20;119(1-2):116-25. Epub 2007 Aug 19.

Climatic models to predict occurrence of Fusarium toxins in wheat and maize.

Schaafsma AW, Hooker DC.

University of Guelph, Ridgetown Campus, Ridgetown, Ontario, Canada N0P 2C0.
aschaafs@ridgetownc.uoguelph.ca

Although forecasting Fusarium infections have useful implications, it may be
argued that forecasting Fusarium toxins is more useful to help reduce their entry
into the food chain. Several disease incidence models have been commercialized
for wheat, but only one toxin prediction model from Ontario, Canada, "DONcast",
has been validated extensively and commercialized to date for wheat, and another
has been proposed for maize. In the development of these predictive tools, the
variation in toxin levels associated with year and agronomic effects was
estimated from simple linear models using wheat and maize samples taken from farm
fields. In wheat, environment effects accounted for 48% of the variation in
deoxynivalenol (DON) across all fields, followed by variety (27%), and previous
crop (14 to 28%). In maize, hybrid accounted for 25% of the variation of either
DON or fumonisin, followed by environment (12%), and when combined 42% of the
variability was accounted for. The robust site-specific, DON forecast model
accounted for up to 80% of the variation in DON, and has been used commercially
for 5 years in Canada. Forecasting DON and fumonisins in maize is more difficult,
because of its greater exposure to infection, the role of wounding in infection,
the more important role of hybrid susceptibility, and the vast array of
uncharacterized hybrids available in the marketplace. Nevertheless, using data
collected from controlled experiments conducted in Argentina and the Philippines,
a model was developed to predict fumonisin concentration using insect damage and
weather variables, accounting for 82% of the variability of fumonisins. Using
mycotoxins as a measure of disease outcome, as opposed to disease symptoms,
offers a more robust prediction of mycotoxin risk, and it accounts for mycotoxin
accumulation that occurs frequently in the absence of any change in Fusarium
symptoms.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17900733 [PubMed - indexed for MEDLINE]

278: Adv Food Nutr Res. 2007;53:161-98.

Designer milk.

Sabikhi L.

Dairy Technology Division, National Dairy Research Institute, Karnal 132001,
Haryana, India.

Dairy biotechnology is fast gaining ground in the area of altering milk
composition for processing and/or animal and human health by employing
nutritional and genetic approaches. Modification of the primary structure of
casein, alteration in the lipid profile, increased protein recovery, milk
containing nutraceuticals, and replacement for infant formula offer several
advantages in the area of processing. Less fat in milk, altered fatty acid
profiles to include more healthy fatty acids such as CLA and omega-fats, improved
amino acid profiles, more protein, less lactose, and absence of
beta-lactoglobulin (beta-LG) are some opportunities of "designing" milk for human
health benefits. Transgenic technology has also produced farm animals that
secrete in their milk, human lactoferrin, lysozyme, and lipase so as to simulate
human milk in terms of quality and quantity of these elements that are protective
to infants. Cow milk allergenicity in children could be reduced by eliminating
the beta-LG gene from bovines. Animals that produce milk containing therapeutic
agents such as insulin, plasma proteins, drugs, and vaccines for human health
have been genetically engineered. In order to cater to animal health, transgenic
animals that express in their mammary glands, various components that work
against mastitis have been generated. The ultimate acceptability of the
"designer" products will depend on ethical issues such as animal welfare and
safety, besides better health benefits and increased profitability of products
manufactured by the novel techniques.

Publication Types:
Review

PMID: 17900499 [PubMed - indexed for MEDLINE]

279: Plant Mol Biol. 2007 Dec;65(6):711-8. Epub 2007 Sep 25.

Systemic wound signaling in tomato leaves is cooperatively regulated by systemin
and hydroxyproline-rich glycopeptide signals.

Narvö¡ez-Vö¡squez J, Orozco-Cö¡rdenas ML, Ryan CA.

Institute of Biological Chemistry, Washington State University, Pullman, WA
99164-6340, USA.

Hydroxyproline-rich glycopeptides (HypSys peptides) have been isolated recently
from tobacco and tomato leaves that are powerful activators of protease inhibitor
synthesis. The peptides are processed from polyprotein precursors, two from a
single tobacco precursor and three from a single tomato precursor. The precursor
genes are expressed in response to wounding and methyl jasmonate, similar to the
expression of the systemin precursor prosystemin in tomato leaves. Here we
investigate the relationships between systemin and the tomato HypSys peptides in
regulating wound signaling in tomato plants. Analysis of transgenic tomato plants
over-expressing sense and antisense constructs of the tomato HypSys precursor
under the 35S CaMV promoter show that the transgenic plants regulate protease
inhibitor gene expression in response to wounding in a manner similar to
prosystemin. The evidence indicates that the expression of both the tomato HypSys
precursor gene and the prosystemin gene in response to wounding are necessary for
strong systemic signaling. The data supports a role for both genes in an
amplification loop that up-regulates the octadecanoid pathway and the synthesis
of jasmonates to effect strong systemic signaling of defense genes. This report
provides the first demonstration of the involvement of two plant peptides derived
from two unrelated genes in regulating long distance wound signaling in plants.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17899396 [PubMed - indexed for MEDLINE]

280: Planta. 2008 Jan;227(2):387-96. Epub 2007 Sep 26.

Functional characterization of the pollen-specific SBgLR promoter from potato
(Solanum tuberosum L.).

Lang Z, Zhou P, Yu J, Ao G, Zhao Q.

State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China
Agricultural University, Beijing, 100094, People's Republic of China.

SBgLR (Solanum tuberosum genomic lysine-rich) gene was isolated from a potato
genomic library using SB401 (S. berthaultii 401) cDNA as probe. RT-PCR analysis
of SBgLR gene expression profile and microscopic analysis of green fluorescent
protein (GFP) expression in tobacco plants transformed with SBgLR promoter-GFP
reporters indicate that SBgLR is a pollen-specific gene. A series of 5'deletions
of SBgLR promoter were fused to the beta-glucuronidase (GUS) gene and stably
introduced into tobacco plants. Histochemical and quantitative assays of GUS
expression in transgenic plants allowed us to localize an enhancer of SBgLR
promoter to the region -345 to -269 relative to the translation start site. This
76 bp (-345 to -269) fragment enhanced GUS expression in leaves, stems and roots
when fused to -90/+6 CaMV 35S minimal promoter. Deletion analysis showed that a
cis-element, which can repress gene expression in root hairs, was located in the
region -345 to -311. Further study indicated that the -269 to -9 region was
sufficient to confer pollen-specific expression of GFP when fused to CaMV 35S
enhancer.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17899173 [PubMed - indexed for MEDLINE]

281: Planta. 2008 Jan;227(2):409-25. Epub 2007 Sep 27.

Distinct roles of the pepper hypersensitive induced reaction protein gene CaHIR1
in disease and osmotic stress, as determined by comparative transcriptome and
proteome analyses.

Jung HW, Lim CW, Lee SC, Choi HW, Hwang CH, Hwang BK.

Laboratory of Molecular Plant Pathology, College of Life Sciences and
Biotechnology, Korea University, Anam-dong, Sungbuk-ku, Seoul, 136-713, South
Korea.

A Capsicum annuum hypersensitive induced reaction protein1 (CaHIR1) was recently
proposed as a positive regulator of hypersensitive cell death in plants.
Overexpression of CaHIR1 in transgenic Arabidopsis plants conferred enhanced
resistance against the hemi-biotrophic Pseudomonas syringae pv. tomato (Pst) and
the biotrophic Hyaloperonospora parasitica. Infection by avirulent Pseudomonas
strains carrying avrRpm1 or avrRpt2 caused enhanced resistance responses in
transgenic plants, suggesting that CaHIR1 is involved in basal disease resistance
in a race-nonspecific manner. H. parasitica exhibited low levels of asexual
sporulation on CaHIR1 seedlings. In contrast, transgenic plants were susceptible
not only to the necrotrophic fungal pathogen Botrytis cinerea but were also
sensitive to osmotic stress caused by high salinity and drought. To identify
proteins whose expression was altered by CaHIR1 overexpression in Arabidopsis
leaves, a quantitative comparative proteome analysis using two-dimensional gel
electrophoresis coupled with mass spectrometry was performed. Of about 400
soluble proteins, 11 proteins involved in several metabolic pathways were up- or
down-regulated by CaHIR1 overexpression. Genes encoding glycine decarboxylase
(At2g35370) and an unidentified protein (At2g03440), which were strongly
upregulated in CaHIR1-overexpressing Arabidopsis, were also differentially
induced at the transcriptional level by Pst infection. Arabidopsis carbonic
anhydrase (At3g01500), highly similar to tobacco salicylic acid-binding protein
3, was up-regulated by CaHIR1 overexpression. The activity of an anti-oxidant
enzyme, cooper/zinc superoxide dismutase (At2g28190), was also attenuated in
transgenic Arabidopsis by CaHIR1 overexpression. Together, these results suggest
that CaHIR1 overexpression in Arabidopsis mediates plant responses to biotrophic,
hemi-biotrophic and necrotrophic pathogens, as well as to osmotic stress in
different ways.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17899171 [PubMed - indexed for MEDLINE]

282: Food Chem Toxicol. 2007 Dec;45(12):2372-80. Epub 2007 Aug 23.

Proteomic analysis of ovomucoid hypersensitivity in mice by two-dimensional
difference gel electrophoresis (2D-DIGE).

Hobson DJ, Rupa P, Diaz GJ, Zhang H, Yang M, Mine Y, Turner PV, Kirby GM.

Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada N1G
2W1.

There is a need to develop reliable methods to assess the safety of genetically
modified and other novel foods. The aim of this study was to identify protein
biomarkers of food allergy in mice exposed to ovomucoid (OVM), a major food
allergen found in chicken egg white. BALB/c mice were repeatedly sensitized by
gavage with OVM and cholera toxin (CT) and control mice were exposed to a mixture
of amino acids with CT. At the endpoint, all mice were challenged
intraperitoneally with OVM and alum. Type-1 hypersensitivity was confirmed in
OVM-sensitized mice by observation of clinical signs of anaphylaxis and elevated
levels of plasma histamine, OVM-specific IgE and OVM-specific IgG by ELISA.
Differential protein expression was assessed in albumin-depleted plasma as well
as in mesenteric lymph node, liver, spleen, and ileum by two-dimensional
difference gel electrophoresis (2D-DIGE). Differentially expressed proteins were
identified by liquid chromatography with tandem mass spectrometry. Plasma
proteins overexpressed in OVM-sensitized mice included haptoglobin (41-fold),
serum amyloid A (19-fold) and peroxiredoxin-2 (1.9-fold). Further validation of
these plasma proteins in other animal models of food allergy with different food
allergens is required to assess their potential as candidate biomarkers for use
in evaluating the allergenicity of novel foods.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17897766 [PubMed - indexed for MEDLINE]

283: Health Res Policy Syst. 2007 Sep 26;5:10.

Biofortification in China: policy and practice.

Campos-Bowers MH, Wittenmyer BF.

Department of Health Management and Policy, School of Public Health, University
of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth,
Texas 76107, USA. mcampos@hsc.unt.edu.

ABSTRACT: Micronutrient deficiency undernutrition, due to insufficient levels of
vitamins and minerals in the diet, remains one of the most prevalent and
preventable nutritional problems in the world today. Micronutrient undernutrition
is the most common form of malnutrition. Compared to the 180 million children
with protein-energy malnutrition, 3.5-5 billion persons are iron-deficient, and
140-250 million persons are vitamin A-deficient. Micronutrient deficiencies
diminish physical, cognitive, and reproductive development. Undernutrition is
both a cause and a result of poor human health and achievement.Middle-income
nations, such as China, also suffer from micronutrient undernutrition's effects.
In China's poor western provinces, despite supplementation and fortification
efforts, stunting and underweight (symptoms of micronutrient undernutrition)
remain common. In recent decades, nutritional adequacy, in terms of available
food energy, improved immensely, as the government made food security a top
priority. A potential next step for China could be to address specifically
micronutrient undernutrition. The paper aims to provide a discussion of policy
issues relevant to biofortification, if China were to consider the implementation
of this intervention in its rural provinces.Traditional nutritional interventions
currently employ four main strategies: dietary modification, supplementation,
commercial fortification, and biofortification. Biofortification, a relatively
new technique, involves selectively breeding staple plant varieties to increase
specific nutrient levels in plant tissues. Biofortification has the potential to
provide benefits to humans, plants, and livestock; nourish nutrient-depleted
soils; and help increase crop yields per acre. Biofortification methods include
selective breeding, reducing levels of anti-nutrients, and increasing levels of
substances that promote nutrient absorption.If China were to implement
biofortification programs, with help from government agencies and international
organizations, several policy questions would need to be addressed. The paper
discusses several policy questions that pertain to the relationship between
biofortified and genetically modified crops, human health and safety concerns,
labeling of biofortified crops for consumers, consumer rights, potential
environmental impacts, intellectual property rights, seed disbursement,
government investment, private-sector research, and additional agricultural and
commercial regulations. Biofortification has the potential to help alleviate the
suffering, death, disability, and failure to achieve full human potential that
results from micronutrient undernutrition-related diseases.

Publication Types:
Editorial

PMID: 17897456 [PubMed - in process]

284: Genes Genet Syst. 2007 Aug;82(4):321-7.

Knockout of glutelin genes which form a tandem array with a high level of
homology in rice by gamma irradiation.

Morita R, Kusaba M, Iida S, Nishio T, Nishimura M.

Institute of Radiation Breeding, National Institute of Agrobiological Sciences,
Kamimurata, Hitachi-ohmiya, Japan. ryo622@affrc.go.jp

In the course of evolution, a gene is often duplicated in tandem, resulting in a
functional redundancy. The analysis of function of these genes by raising double
mutant might be difficult because they are very tightly linked. We described here
a mutant of such a tandem duplicated gene. glu1 is a gamma-ray-induced rice
mutant, which lacks an acidic subunit of glutelin, a major seed storage protein.
We found that glu1 harbors a 129.7-kb deletion involving two highly similar and
tandem repeated glutelin genes, GluB5 and GluB4. The deletion eliminated the
entire GluB5 and GluB4 gene except half of the first exon of GluB5. GluB5 and
GluB4 have the same amino acid sequence in the acidic subunit, suggesting that
only the mutation involving both GluB5 and GluB4 results in the lack of the
glutelin acidic subunit deleted in glu1. Our finding suggests that gamma-ray can
be an effective mutagen to analyze tandem repeated and functionally redundant
genes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17895583 [PubMed - indexed for MEDLINE]

285: Genome. 2007 Jul;50(7):627-37.

Quantitative trait locus analysis of multiple agronomic traits in the model
legume Lotus japonicus.

Gondo T, Sato S, Okumura K, Tabata S, Akashi R, Isobe S.

University of Miyazaki, Frontier Science Research Center, 1-1 Nishi
Gakuen-Kibanadai, Miyazaki 889-2192, Japan.

The first quantitative trait locus (QTL) analysis of multiple agronomic traits in
the model legume Lotus japonicus was performed with a population of recombinant
inbred lines derived from Miyakojima MG-20 x Gifu B-129. Thirteen agronomic
traits were evaluated in 2004 and 2005: traits of vegetative parts (plant height,
stem thickness, leaf length, leaf width, plant regrowth, plant shape, and stem
color), flowering traits (flowering time and degree), and pod and seed traits
(pod length, pod width, seeds per pod, and seed mass). A total of 40 QTLs were
detected that explained 5%-69% of total variation. The QTL that explained the
most variation was that for stem color, which was detected in the same region of
chromosome 2 in both years. Some QTLs were colocated, especially those for pod
and seed traits. Seed mass QTLs were located at 5 locations that mapped to the
corresponding genomic positions of equivalent QTLs in soybean, pea, chickpea, and
mung bean. This study provides fundamental information for breeding of
agronomically important legume crops.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17893740 [PubMed - indexed for MEDLINE]

286: Rev Sci Tech. 2007 Aug;26(2):461-70.

Vaccines for immunological control of fertility in animals.

Hardy CM, Braid AL.

Commonwealth Scientific and Industrial Research Organisation, GPO Box 1700,
Canberra, ACT, 2601, Australia. chris.hardy@csiro.au

Fertility control has gained considerable momentum as a management tool to
regulate populations of captive and wild animals and to control aggressive
behaviour or improve meat quality in livestock. Anti-fertility vaccination
(immunocontraception and immunocastration) is a humane alternative to methods
that rely on surgical or chemical sterilisation and lethal control. Two types of
experimental immunocontraceptive vaccine have been registered for field use in
animals. They contain either porcine zona pellucida (PZP) proteins extracted from
pig ovaries or synthetic conjugated gonadotrophin releasing hormone (GnRH)
peptides. These vaccines require repeated injections and are limited to captive
or small populations of free-ranging wild animals. Alternative
immunocontraceptive vaccines are actively being developed either to improve
efficacy or enable large numbers of wild animals to be targeted. Some employ live
genetically modified viruses to deliver immunocontraception and have proved
successful under laboratory conditions. The relative merits, risks, social
acceptability and regulations controlling the use of existing and novel animal
immunocontraceptives are reviewed.

Publication Types:
Review

PMID: 17892166 [PubMed - indexed for MEDLINE]

287: Plant Mol Biol. 2007 Nov;65(4):385-402. Epub 2007 Sep 19.

Ds insertion mutagenesis as an efficient tool to produce diverse variations for
rice breeding.

Jiang SY, Bachmann D, La H, Ma Z, Venkatesh PN, Ramamoorthy R, Ramachandran S.

Rice Functional Genomics Group, Temasek Life Sciences Laboratory, 1 Research
Link, National University of Singapore, Singapore, 117604, Singapore.

The availability of diversified germplasm resources is the most important for
developing improved rice varieties with higher seed yield or tolerance to various
biotic or abiotic stresses. Here we report an efficient tool to create increased
variations in rice by maize Ac/Ds transposon (a gene trap system) insertion
mutagenesis. We have generated around 20,000 Ds insertion rice lines of which
majority are homozygous for Ds element. We subjected these lines to phenotypic
and abiotic stress screens and evaluated these lines with respect to their seed
yields and other agronomic traits as well as their tolerance to drought, salinity
and cold. Based on this evaluation, we observed that random Ds insertions into
rice genome have led to diverse variations including a range of morphological and
conditional phenotypes. Such differences in phenotype among these lines were
accompanied by differential gene expression revealed by GUS histochemical
staining of gene trapped lines. Among the various phenotypes identified, some Ds
lines showed significantly higher grain yield compared to wild-type plants under
normal growth conditions indicating that rice could be improved in grain yield by
disrupting certain endogenous genes. In addition, several 1,000s of Ds lines were
subjected to abiotic stresses to identify conditional mutants. Subsequent to
these screens, over 800 lines responsive to drought, salinity or cold stress were
obtained, suggesting that rice has the genetic potential to survive under abiotic
stresses when appropriate endogenous genes were suppressed. The mutant lines that
have higher seed yielding potential or display higher tolerance to abiotic
stresses may be used for rice breeding by conventional backcrossing combining
with molecular marker-assisted selection. In addition, by exploiting the behavior
of Ds to leave footprints upon remobilization, we have shown an alternative
strategy to develop new rice varieties without foreign DNA sequences in their
genome.

PMID: 17891459 [PubMed - indexed for MEDLINE]

288: Plant Cell. 2007 Sep;19(9):2913-28. Epub 2007 Sep 21.

Functional dissection of naturally occurring amino acid substitutions in eIF4E
that confers recessive potyvirus resistance in plants.

Yeam I, Cavatorta JR, Ripoll DR, Kang BC, Jahn MM.

Department of Plant Breeding and Genetics, College of Agriculture and Life
Sciences, Cornell University, Ithaca, NY 14853, USA.

Naturally existing variation in the eukaryotic translation initiation factor 4E
(eIF4E) homolog encoded at the pvr1 locus in Capsicum results in recessively
inherited resistance against several potyviruses. Previously reported data
indicate that the physical interaction between Capsicum-eIF4E and the viral
genome-linked protein (VPg) is required for the viral infection in the
Capsicum-Tobacco etch virus (TEV) pathosystem. In this study, the potential
structural role(s) of natural variation in the eIF4E protein encoded by recessive
resistance alleles and their biological consequences have been assessed. Using
high-resolution three-dimensional structural models based on the available
crystallographic structures of eIF4E, we show that the amino acid substitution
G107R, found in many recessive plant virus resistance genes encoding eIF4E, is
predicted to result in a substantial modification in the protein binding pocket.
The G107R change was shown to not only be responsible for the interruption of VPg
binding in planta but also for the loss of cap binding ability in vitro, the
principal function of eIF4E in the host. Overexpression of the Capsicum-eIF4E
protein containing the G107R amino acid substitution in Solanum lycopersicum
indicated that this polymorphism alone is sufficient for the acquisition of
resistance against several TEV strains.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17890375 [PubMed - indexed for MEDLINE]

289: New Phytol. 2007;176(2):288-98.

Overexpression of a NAC-domain protein promotes shoot branching in rice.

Mao C, Ding W, Wu Y, Yu J, He X, Shou H, Wu P.

State Key Laboratory of Plant Physiology and Biochemistry, College of Life
Sciences, Zhejiang University, Hangzhou, 310058, PR China.

For a better understanding of shoot branching in rice (Oryza sativa), a rice
activation-tagging library was screened for mutations in tiller development.
Here, an activation-tagging mutant Ostil1 (Oryza sativa tillering1) was
characterized, which showed increased tillers, enlarged tiller angle and
semidwarf phenotype. Flanking sequence was obtained by plasmid rescue.
RNA-interfering and overexpression transgenic rice plants were produced using
Agrobacterium-mediated transformation. The mutant phenotype was cosegregated with
the reallocation of Ds element, and the flanking region of the reallocated Ds
element was identified as part of the OsNAC2 gene. Northern analysis showed that
expression of OsNAC2 was greatly induced in the mutant plants. Transgenic rice
overexpressing the OsNAC2 resulted in recapture of the mutant phenotype, while
downregulation of OsNAC2 in the Ostil1 mutant through RNA interfering (RNAi)
complemented the mutant phenotype, confirming that the Ostil1 was caused by
overexpression of OsNAC2. Overexpression of OsNAC2 regulates shoot branching in
rice. Overexpression of OsNAC2 contributes tiller bud outgrowth, but does not
affect tiller bud initiation. This suggests that OsNAC2 has potential utility for
improving plant structure for higher light-use efficiency and higher yield
potential in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17888111 [PubMed - indexed for MEDLINE]

290: Plant J. 2007 Aug;51(4):575-88. Epub 2007 Jun 6.

Members of the plant NIMA-related kinases are involved in organ development and
vascularization in poplar, Arabidopsis and rice.

Vigneault F, Lachance D, Cloutier M, Pelletier G, Levasseur C, Sö©guin A.

Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre,
1055 du P.E.P.S., PO Box 10380, Stn. Sainte-Foy, Quebec, QC, Canada G1 V 4C7.

NIMA-related kinases (Neks) are a family of serine/threonine kinases that have
been linked to cell-cycle regulation in fungi and mammals. Information regarding
the function of Neks in plants is very limited. We screened the three plant
species that have had their genomes sequenced in an attempt to improve our
understanding of their role in plants. We retrieved seven members in Arabidopsis
thaliana, nine in Populus trichocarpa and six in Oryza sativa. Phylogenetic
analysis showed that plant Neks are closely related to each other and contain
paralogous genes. Moreover, their chromosome distribution and their exon-intron
structure revealed that the actual plant Nek family was derived from a single
representative followed by large segmental duplication events. Functional
expression analyses in the three species relied on RTqPCR in poplar and publicly
available microarray data for Arabidopsis and rice. Although plant Neks are
present in every organ analyzed, their expression profiles suggest their
involvement in plant development processes. Furthermore, we showed that PNek1, a
member of the poplar family, is expressed at sites of free auxin synthesis and is
specifically involved during the vascularization process.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17886359 [PubMed - indexed for MEDLINE]

291: Plant Physiol. 2007 Nov;145(3):626-39. Epub 2007 Sep 20.

Reduced expression of succinyl-coenzyme A ligase can be compensated for by
up-regulation of the gamma-aminobutyrate shunt in illuminated tomato leaves.

Studart-Guimarö¸es C, Fait A, Nunes-Nesi A, Carrari F, Usadel B, Fernie AR.

Department Willmitzer, Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie,
14476 Golm, Germany.

Increasing experimental evidence suggests that the tricarboxylic acid cycle in
plants is of greater importance in illuminated photosynthetic tissues than
previously thought. In this study, transgenic tomato (Solanum lycopersicum)
plants expressing a fragment of the beta-subunit of succinyl-coenzyme A ligase in
either the antisense orientation or using the RNA interference approach, however,
revealed little alteration in either photosynthesis or plant growth despite
exhibiting dramatic reductions in activity. Moreover, the rate of respiration was
only moderately affected in the transformants, suggesting that this enzyme does
not catalyze a crucial step in mitochondrial respiration. However, metabolite and
transcript profiling of these lines alongside enzyme and label redistribution
experiments revealed that, whereas considerable activity of this enzyme appears
to be dispensable, the reason for such a mild phenotype in extremely inhibited
lines was an up-regulation of an alternative pathway for succinate
production-that offered by the gamma-aminobutyric acid shunt. When taken
together, these data highlight the importance both of succinate production for
mitochondrial metabolism and the interplay between various routes of its
production. The results are discussed in the context of current models of plant
respiration in mitochondrial and cellular metabolism of the illuminated leaf.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17885090 [PubMed - indexed for MEDLINE]

292: Plant Physiol. 2007 Nov;145(3):640-52. Epub 2007 Sep 20.

Alteration of organic acid metabolism in Arabidopsis overexpressing the maize C4
NADP-malic enzyme causes accelerated senescence during extended darkness.

Fahnenstich H, Saigo M, Niessen M, Zanor MI, Andreo CS, Fernie AR, Drincovich MF,
Flö¼gge UI, Maurino VG.

Botanisches Institut, Universitö¤t zu Kö¶ln, 50931 Cologne, Germany.

The full-length cDNA encoding the maize (Zea mays) C(4) NADP-malic enzyme was
expressed in Arabidopsis (Arabidopsis thaliana) under the control of the
cauliflower mosaic virus 35S promoter. Homozygous transgenic plants (MEm) were
isolated with activities ranging from 6- to 33-fold of those found in the wild
type. The transformants did not show any differences in morphology and
development when grown in long days; however, dark-induced senescence progressed
more rapidly in MEm plants compared to the wild type. Interestingly, senescence
could be retarded in the transgenic lines by exogenously supplying glucose,
sucrose, or malate, suggesting that the lack of a readily mobilized carbon source
is likely to be the initial factor leading to the premature induction of
senescence in MEm plants. A comprehensive metabolic profiling on whole rosettes
allowed determination of approximately 80 metabolites during a diurnal cycle as
well as following dark-induced senescence and during metabolic complementation
assays. MEm plants showed no differences in the accumulation and degradation of
carbohydrates with respect to the wild type in all conditions tested, but
accumulated lower levels of intermediates used as respiratory substrates,
prominently malate and fumarate. The data indicated that extremely low levels of
malate and fumarate are responsible for the accelerated dark-induced senescence
encountered in MEm plants. Thus, in prolonged darkness these metabolites are
consumed faster than in the wild type and, as a consequence, MEm plants enter
irreversible senescence more rapidly. In addition, the data revealed that both
malate and fumarate are important forms of fixed carbon that can be rapidly
metabolized under stress conditions in Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17885087 [PubMed - indexed for MEDLINE]

293: Plant Physiol. 2007 Nov;145(3):946-60. Epub 2007 Sep 20.

Localization in roots and flowers of pea chloroplastic thioredoxin f and
thioredoxin m proteins reveals new roles in nonphotosynthetic organs.

de Dios Barajas-Lö¨pez J, Serrato AJ, Olmedilla A, Chueca A, Sahrawy M.

Estaciö¨n Experimental del Zaidö­n, Consejo Superior de Investigaciones
Cientö­ficas, 18008 Granada, Spain.

Plant thioredoxins (TRXs) are involved in redox regulation of a wide variety
processes and usually exhibit organ specificity. We report strong evidence that
chloroplastic TRXs are localized in heterotrophic tissues and suggest some ways
in which they might participate in several metabolic and developmental processes.
The promoter regions of the chloroplastic f and m1 TRX genes were isolated from a
pea (Pisum sativum) plant genomic bank. Histochemical staining for
beta-glucuronidase (GUS) in transgenic homozygous Arabidopsis (Arabidopsis
thaliana) plants showed preferential expression of the 444-bp PsTRXf1 promoter in
early seedlings, stems, leaves, and roots, as well as in flowers, stigma, pollen
grains, and filaments. GUS activity under the control of the 1,874-bp PsTRXm1
promoter was restricted to the leaves, roots, seeds, and flowers. To gain insight
into the translational regulation of these genes, a series of deletions of 5'
elements in both TRX promoters were analyzed. The results revealed that a 126-bp
construct of the PsTRXf2 promoter was unable to reproduce the expression pattern
observed with the full promoter. The differences in expression and tissue
specificity between PsTRXm1 and the deleted promoters PsTRXm2 and PsTRXm3 suggest
the existence of upstream positive or negative regulatory regions that affect
tissue specificity, sucrose metabolism, and light regulation. PsTRXm1 expression
is finely regulated by light and possibly by other metabolic factors. In situ
hybridization experiments confirmed new localizations of these chloroplastic TRX
transcripts in vascular tissues and flowers, and therefore suggest possible new
functions in heterotrophic tissues related to cell division, germination, and
plant reproduction.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17885084 [PubMed - indexed for MEDLINE]

294: Plant Physiol. 2007 Nov;145(3):601-15. Epub 2007 Sep 20.

Early steps in proanthocyanidin biosynthesis in the model legume Medicago
truncatula.

Pang Y, Peel GJ, Wright E, Wang Z, Dixon RA.

Plant Biology Division , Samuel Roberts Noble Foundation, Ardmore, Oklahoma
73401, USA.

Oligomeric proanthocyanidins (PAs) composed primarily of epicatechin units
accumulate in the seed coats of the model legume Medicago truncatula, reaching
maximal levels at around 20 d after pollination. Genes encoding the single
Medicago anthocyanidin synthase (ANS; EC 1.14.11.19) and leucoanthocyanidin
reductase (LAR; EC 1.17.1.3) were cloned and the corresponding enzymes
functionally identified. Recombinant MtANS converted leucocyanidin to cyanidin,
and, more efficiently, dihydroquercetin to the flavonol quercetin. Levels of
transcripts encoding dihydroflavonol reductase, ANS, and anthocyanidin reductase
(ANR), the enzyme responsible for conversion of anthocyanidin to (-)-epicatechin,
paralleled the accumulation of PAs in developing seeds, whereas LAR transcripts
appeared to be more transiently expressed. LAR, ANS, and ANR proteins were
localized to the cytosol in transfected tobacco (Nicotiana tabacum) leaves.
Antisense down-regulation of ANS in M. truncatula resulted in reduced anthocyanin
and PA levels, but had no impact on flavonol levels. Transgenic tobacco plants
constitutively overexpressing MtLAR showed reduced anthocyanin content, but no
catechin or increased levels of PAs were detected either in leaves or in flowers.
Our results confirm previously ascribed in vivo functions for ANS and ANR.
However, the apparent lack of catechin in M. truncatula PAs, the poor correlation
between LAR expression and PA accumulation, and the lack of production of
catechin monomers or oligomers in transgenic plants overexpressing MtLAR question
the role of MtLAR in PA biosynthesis in Medicago.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17885080 [PubMed - indexed for MEDLINE]

295: Plant Mol Biol. 2007 Dec;65(6):733-46. Epub 2007 Sep 20.

Overexpression of a putative maize calcineurin B-like protein in Arabidopsis
confers salt tolerance.

Wang M, Gu D, Liu T, Wang Z, Guo X, Hou W, Bai Y, Chen X, Wang G.

State Key Laboratory of Agrobiotechnology and National Center for Plant Gene
Research (Beijing), China Agricultural University, Beijing 100094, China.

The calcineurin B-like proteins (CBLs) represent a unique family of calcium
sensors in plants. Although extensive studies and remarkable progress have been
made in Arabidopsis (Arabidopsis thaliana) CBLs, their functions in other plant
species are still quite limited. Here, we report the cloning and functional
characterization of ZmCBL4, a novel CBL gene from maize (Zea mays). ZmCBL4
encodes a putative homolog of the Arabidopsis CBL4/SOS3 protein, with novel
properties. ZmCBL4 has one copy in maize genome and harbors seven introns in its
coding region. ZmCBL4 expressed differentially in various organs of the maize
plants at a low level under normal condition, and its expression was regulated by
NaCl, LiCl, ABA and PEG treatments. Expression of 35S::ZmCBL4 not only
complemented the salt hypersensitivity in Arabidopsis sos3 mutant, but also
enhanced the salt tolerance in Arabidopsis wild type at the germination and
seedling stages. Moreover, the LiCl tolerance in all of the ZmCBL4-expressing
lines increased more significantly as compared with the NaCl tolerance, and in
consistent with this, it was found that the expression of Arabidopsis AtNHX8, a
putative plasma membrane Li+/H+ antiporter gene identified recently, was induced
in these transgenic lines under LiCl stress. The ZmCBL4-expressing Arabidopsis
lines accumulated less Na+ and Li+ as compared with the control plants. This
study has identified a putative maize CBL gene which functions in the salt
stress-elicited calcium signaling and thus in the tolerance to salinity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17882512 [PubMed - indexed for MEDLINE]

296: Pest Manag Sci. 2007 Nov;63(11):1107-15.

Altered pesticide use on transgenic crops and the associated general impact from
an environmental perspective.

Kleter GA, Bhula R, Bodnaruk K, Carazo E, Felsot AS, Harris CA, Katayama A,
Kuiper HA, Racke KD, Rubin B, Shevah Y, Stephenson GR, Tanaka K, Unsworth J,
Wauchope RD, Wong SS.

RIKILT-Institute of Food Safety, Wageningen University and Research Centre,
Wageningen, Holland. gijs.kleter@wur.nl

The large-scale commercial cultivation of transgenic crops has undergone a steady
increase since their introduction 10 years ago. Most of these crops bear
introduced traits that are of agronomic importance, such as herbicide or insect
resistance. These traits are likely to impact upon the use of pesticides on these
crops, as well as the pesticide market as a whole. Organizations like USDA-ERS
and NCFAP monitor the changes in crop pest management associated with the
adoption of transgenic crops. As part of an IUPAC project on this topic, recent
data are reviewed regarding the alterations in pesticide use that have been
observed in practice. Most results indicate a decrease in the amounts of active
ingredients applied to transgenic crops compared with conventional crops. In
addition, a generic environmental indicator -- the environmental impact quotient
(EIQ) -- has been applied by these authors and others to estimate the
environmental consequences of the altered pesticide use on transgenic crops. The
results show that the predicted environmental impact decreases in transgenic
crops. With the advent of new types of agronomic trait and crops that have been
genetically modified, it is useful to take also their potential environmental
impacts into account.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17880042 [PubMed - indexed for MEDLINE]

297: Sci China C Life Sci. 2007 Oct;50(5):573-9.

Progress in the evaluation of transgenic fish for possible ecological risk and
its containment strategies.

Hu W, Wang Y, Zhu Z.

State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of
Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.

Genetically improved transgenic fish possess many beneficial economic traits;
however, the commercial aquaculture of transgenic fish has not been performed
till date. One of the major reasons for this is the possible ecological risk
associated with the escape or release of the transgenic fish. Using a growth
hormone transgenic fish with rapid growth characteristics as a subject, this
paper analyzes the following: the essence of the potential ecological risks posed
by transgenic fish; ecological risk in the current situation due to transgenic
fish via one-factor phenotypic and fitness analysis, and mathematical model
deduction. Then, it expounds new ideas and the latest findings using an
artificially simulated ecosystem for the evaluation of the ecological risks posed
by transgenic fish. Further, the study comments on the strategies and principles
of controlling these ecological risks by using a triploid approach. Based on
these results, we propose that ecological risk evaluation and prevention
strategies are indispensable important components and should be accompanied with
breeding research in order to provide enlightments for transgenic fish breeding,
evaluation of the ecological risks posed by transgenic fish, and development of
containment strategies against the risks.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17879053 [PubMed - indexed for MEDLINE]

298: Plant J. 2007 Nov;52(4):627-39. Epub 2007 Sep 18.

The identification of a gene (Cwp1), silenced during Solanum evolution, which
causes cuticle microfissuring and dehydration when expressed in tomato fruit.

Hovav R, Chehanovsky N, Moy M, Jetter R, Schaffer AA.

Institute of Field and Garden Crops, ARO, The Volcani Center, Bet Dagan 50250,
Israel.

One of the most intriguing phenomena of fleshy fruit is the ability to maintain
high water content at maturity, even following harvest. This is accomplished by a
fruit cuticle that is highly impermeable to water diffusion. In this paper, we
report on a novel genotype of tomato, developed via introgression from the wild
species Solanum habrochaites, which is characterized by microfissuring of the
fruit cuticle and dehydration of the mature fruit. The microfissure/dehydration
phenotype is inherited as a single gene, termed Cwp1 (cuticular water
permeability). The gene was fine mapped, and its identity was determined by
map-based cloning and differential expression analysis in near-isogenic lines.
Causality of the Cwp1 gene was shown by the heterologous transgenic expression of
the gene in the cultivated tomato, which caused a microfissured fruit cuticle
leading to dehydrated fruit. Cwp1 encodes for a protein of unidentified function
in the DUF833 domain family. The gene is expressed in the fruit epidermis of the
dehydrating genotype harbouring the wild-species introgression, but not in the
cultivated tomato. It is expressed only in the primitive green-fruited wild
tomato species, but is not expressed in the cultivated Solanum lycopersicum and
the closely related Solanum cheesmaniae and Solanum pimpinellifolium, indicating
a pre-adaptive role for Cwp1 silencing in the evolution and domestication of the
cultivated tomato.

PMID: 17877702 [PubMed - indexed for MEDLINE]

299: Plant J. 2007 Nov;52(4):716-29. Epub 2007 Sep 18.

The soybean Dof-type transcription factor genes, GmDof4 and GmDof11, enhance
lipid content in the seeds of transgenic Arabidopsis plants.

Wang HW, Zhang B, Hao YJ, Huang J, Tian AG, Liao Y, Zhang JS, Chen SY.

National Key Laboratory of Plant Genomics, Institute of Genetics and
Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.

Soybean is one of the most important leguminous seed crops among the oil crops.
Although the pathways for lipid biosynthesis have been identified, the factors
that regulate the biosynthetic pathways at the transcriptional level are largely
unknown. Here, we report our findings on the involvement of soybean Dof-type
transcription factor genes in the regulation of the lipid content in soybean
seeds. We identified 28 Dof-type transcription factor genes in soybean plants,
and these genes displayed diverse patterns of expression in various organs. Seven
flower/pod-specific genes and one constitutively expressed gene were further
investigated. The proteins encoded by these seven genes were localized in the
nucleus, and exhibited different abilities for transcriptional activation and DNA
binding. Two genes, GmDof4 and GmDof11, were found to increase the content of
total fatty acids and lipids in GmDof4 and GmDof11 transgenic Arabidopsis seeds.
We also found that the 1000-seed weight was increased in the GmDof4 and GmDof11
transgenic plants. Using microarray and DNA binding analysis, we found that the
two Dof-like proteins, GmDof4 and GmDof11, activated the acetyl CoA carboxylase
gene and long-chain-acyl CoA synthetase gene, respectively, by direct binding to
the cis-DNA elements in their promoter regions. In addition, both proteins
downregulated the storage protein gene, CRA1, through direct binding. These
results suggest that the two GmDof genes may augment the lipid content of soybean
seeds by upregulating genes that are associated with the biosynthesis of fatty
acids.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17877700 [PubMed - indexed for MEDLINE]

300: Plant J. 2007 Nov;52(4):752-62. Epub 2007 Sep 18.

Geranyl diphosphate synthase is required for biosynthesis of gibberellins.

van Schie CC, Ament K, Schmidt A, Lange T, Haring MA, Schuurink RC.

Department of Plant Physiology, Swammerdam Institute for Life Sciences,
University of Amsterdam, 1098 SM Amsterdam, The Netherlands. cvanschie@ucsd.edu

Geranyl diphosphate synthase (GPS) is generally considered to be responsible for
the biosynthesis of monoterpene precursors only. However, reduction of LeGPS
expression in tomato (Lycopersicon esculentum) by virus-induced gene silencing
resulted in severely dwarfed plants. Further analysis of these dwarfed plants
revealed a decreased gibberellin content, whereas carotenoid and chlorophyll
levels were unaltered. Accordingly, the phenotype could be rescued by application
of gibberellic acid. The dwarfed phenotype was also obtained in Arabidopsis
thaliana plants transformed with RNAi constructs of AtGPS. These results link
geranyl diphosphate (GPP) to the gibberellin biosynthesis pathway. They also
demand a re-evaluation of the role of GPS in precursor synthesis for other di-,
tri-, tetra- and/or polyterpenes and their derivatives.

PMID: 17877699 [PubMed - indexed for MEDLINE]

301: Cell Res. 2007 Oct;17(10):881-94.

Genome-wide analysis of the phospholipase D family in Oryza sativa and functional
characterization of PLD beta 1 in seed germination.

Li G, Lin F, Xue HW.

National Key Laboratory of Plant Molecular Genetics, Institute of Plant
Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.

Phospholipase D (PLD) plays a critical role in plant growth and development, as
well as in hormone and stress responses. PLD encoding genes constitute a large
gene family that are present in higher plants. There are 12 members of the PLD
family in Arabidopsis thaliana and several of them have been functionally
characterized; however, the members of the PLD family in Oryza sativa remain to
be fully described. Through genome-wide analysis, 17 PLD members found in
different chromosomes have been identified in rice. Protein domain structural
analysis reveals a novel subfamily, besides the C2-PLDs and PXPH-PLDs, that is
present in rice - the SP-PLD. SP-PLD harbors a signal peptide instead of the C2
or PXPH domains at the N-terminus. Expression pattern analysis indicates that
most PLD-encoding genes are differentially expressed in various tissues, or are
induced by hormones or stress conditions, suggesting the involvement of PLD in
multiple developmental processes. Transgenic studies have shown that the
suppressed expression of rice PLD beta 1 results in reduced sensitivity to
exogenous ABA during seed germination. Further analysis of the expression of ABA
signaling-related genes has revealed that PLD beta 1 stimulates ABA signaling by
activating SAPK, thus repressing GAmyb expression and inhibiting seed
germination.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17876344 [PubMed - indexed for MEDLINE]

302: Plant Mol Biol. 2007 Dec;65(6):719-32. Epub 2007 Sep 15.

Isolation and molecular characterization of the Triticum aestivum L.
ethylene-responsive factor 1 (TaERF1) that increases multiple stress tolerance.

Xu ZS, Xia LQ, Chen M, Cheng XG, Zhang RY, Li LC, Zhao YX, Lu Y, Ni ZY, Liu L,
Qiu ZG, Ma YZ.

National Key Facility of Crop Gene Resources and Genetic Improvement, Key
Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Institute of
Crop Science, Chinese Academy of Agriculture Sciences, Beijing 100081, PR China.

ERF transcription factors play important roles in regulating gene expression
under abiotic and biotic stresses. The first member of the ERF gene family in
wheat (Triticum aestivum L.) was isolated by screening a drought-induced cDNA
library and designated as T. aestivum ethylene-responsive factor 1 (TaERF1),
which encoded a putative protein of 355 amino acids with a conserved DNA-binding
domain and a conserved N-terminal motif (MCGGAIL). The TaERF1 gene was located on
chromosome 7A. Protein interaction assays indicated that TaERF1, with a putative
phosphorylation site (TPDITS) in the C-terminal region, was a potential
phosphorylation substrate for TaMAPK1 protein kinase. Deletion of the N-terminal
motif enhanced the interaction of TaERF1 with TaMAPK1. The predicted TaERF1
protein contained three putative nuclear localization signals (NLSs), and three
NLSs modulated synergistically the activity of subcellular localization. As a
trans-acting factor, TaERF1 was capable of binding to the GCC-box and CRT/DRE
elements in vitro, and of trans-activating reporter gene expression in tobacco
(Nicotiana tabacum L.) leaves. Transcription of the TaERF1 gene was induced not
only by drought, salinity and low-temperature stresses and exogenous ABA,
ethylene and salicylic acid, but also by infection with Blumeria graminis f. sp.
tritici. Furthermore, overexpression of TaERF1 activated stress-related genes,
including PR and COR/RD genes, under normal growth conditions, and improved
pathogen and abiotic stress tolerance in transgenic plants. These results
suggested that the TaERF1 gene encodes a GCC-box and CRT/DRE element binding
factor that might be involved in multiple stress signal transduction pathways.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17874224 [PubMed - indexed for MEDLINE]

303: Biotechnol Appl Biochem. 2007 Oct;48(Pt 2):101-7.

Expression of human coagulation Factor IX in transgenic tomato (Lycopersicon
esculentum).

Zhang H, Zhao L, Chen Y, Cui L, Ren W, Tang K.

Plant Biotechnology Research Center, School of Agriculture and Biology,
Fudan-Shanghai Jiaotong University (SJTU)-Nottingham Plant Biotechnology R&D
Center, SJTU, Shanghai 200030, People's Republic of China.

In the present study, a plant binary expression vector PG-pRD12-hFIX (where PG is
polygalacturonase) harbouring the hFIX (human coagulation Factor IX) gene was
constructed and introduced into tomato (Lycopersicon esculentum) via
Agrobacterium tumefaciens-mediated transformation. After kanamycin selection, 32
putative independent transgenic tomato plants were regenerated. PCR and
Southern-blot analyses confirmed the transgenic status of some plants. RT
(reverse transcription)-PCR analysis for the expression of the introduced gene
(hFIX) demonstrated that the hFIX gene was expressed specifically in fruits of
the tomato. Western-blot analysis confirmed the presence of a 56 kDa band
specific to hFIX in the transformed tomatoes. ELISA results showed that the
expression of hFIX protein reached a maximum of 15.84 ng/g fresh weight in mature
fruit. A blood-clotting assay demonstrated the clotting activity of the expressed
hFIX protein in transgenic tomato fruits. This is the first report on the
expression of hFIX in plants, and our research provides potentially valuable
knowledge for further development of the plant-derived therapeutic proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17868024 [PubMed - indexed for MEDLINE]

304: Development. 2007 Oct;134(20):3639-48. Epub 2007 Sep 12.

Polycomb group proteins function in the female gametophyte to determine seed
development in plants.

Leroy O, Hennig L, Breuninger H, Laux T, Kö¶hler C.

Institute of Plant Sciences and Zö¼rich-Base Plant Science Center, Swiss Federal
Institute of Technology, ETH Centre, CH-8092 Zö¼rich, Switzerland.

Polycomb group (PcG) proteins are evolutionary conserved proteins that stably
maintain established transcriptional patterns over cell generations. The
FERTILIZATION INDEPENDENT SEED (FIS) PcG complex from plants has a similar
composition to the Polycomb repressive complex 2 from animals. Mutations in FIS
genes cause parent-of-origin-dependent seed abortion. Every seed inheriting a
mutant fis allele from the mother is destined to abort, regardless of the
presence of a wild-type paternal allele. We tested in Arabidopsis whether the
parent-of-origin-dependent seed abortion caused by lack of the FIS subunit MSI1
is caused by parental imprinting of the MSI1 gene. Our data show that MSI1 is not
an imprinted gene and that early paternal MSI1 expression is not sufficient to
rescue msi1 mutant seeds. By contrast, expression of MSI1 in msi1 female
gametophytes is necessary to restore normal seed development, strongly arguing
that the female gametophytic effect of fis mutants is caused by a functional
requirement for an intact FIS complex in the female gametophyte. Thus,
FIS-mediated expression patterns established in the female gametophyte can impact
on seed development, establishing fis mutants as true female gametophytic
maternal-effect mutants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17855429 [PubMed - indexed for MEDLINE]

305: Med Pregl. 2007 Mar-Apr;60(3-4):195-7.

[Diseases caused by bacteria and rickettsia in biological warfare and
bioterrorism]

[Article in Serbian]

Bojiä‡ I, Vukadinov J, Miniä‡ S.

Specijalisticka ordinacija Dr Bojiä‡, Beograd. drbojic@net.yu

INTRODUCTION: Until recently, the use of biological weapons was considered more
from an academic than practical point of view. The list of agents and/or toxins
that can be used as biological weapons is long. Some of them are highly lethal,
while others cause morbidity and disability. BIOLOGICAL WEAPONS: Bacteria,
rickettsia, viruses, fungi, protozoa and toxins can all be used as biological
weapons. The infection may be acquired by inhalation of aerosols, ingestion of
contaminated water or food or direct contact with infectious agents. Early
recognition, diagnosis and treatment of these patients is of utmost importance.
Special attention must be given to the use of genetically modified
microorganisms. Medical protection from biological weapons is very important as
well as continuous education. CONCLUSION: This article describes the main
clinical characteristics of anthrax, cholera, plague, Q fever, tularemia,
brucellosis, and glanders, as biological weapons, their diagnostics, treatment
and basic prevention measures.

Publication Types:
English Abstract
Review

PMID: 17853736 [PubMed - indexed for MEDLINE]

306: Fortune. 2007 Jul 9;156(1):74-8, 80.

Attack of the mutant rice.

Gunther M.

Publication Types:
News

PMID: 17853593 [PubMed - indexed for MEDLINE]

307: Anal Bioanal Chem. 2007 Oct;389(3):913-21. Epub 2007 Sep 13.

A DNA electrochemical sensor prepared by electrodepositing zirconia on composite
films of single-walled carbon nanotubes and poly(2,6-pyridinedicarboxylic acid),
and its application to detection of the PAT gene fragment.

Yang J, Jiao K, Yang T.

College of Chemistry and Molecular Engineering, Qingdao University of Science and
Technology, Qingdao 266042, China.

Carboxyl group-functionalized single-walled carbon nanotubes (SWNTs) and
2,6-pyridinedicarboxylic acid (PDC) were electropolymerized by cyclic voltammetry
on a glassy-carbon electrode (GCE) surface to form composite films (SWNTs/PDC).
Zirconia was then electrodeposited on the SWNTs/PDC/GCE from an aqueous
electrolyte containing ZrOCl2 and KCl by cycling the potential between -1.1 V and
+0.7 V at a scan rate of 20 mV s(-1). DNA probes with a phosphate group at the 5'
end were easily immobilized on the zirconia thin films, because of the strong
affinity between zirconia and phosphate groups. The sensors were characterized by
cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). EIS was
used for label-free detection of the target DNA by measuring the increase of the
electron transfer resistance (R(et)) of the electrode surface after the
hybridization of the probe DNA with the target DNA. The PAT gene fragment and
polymerase chain reaction (PCR) amplification of the NOS gene from transgenically
modified beans were satisfactorily detected by use of this DNA electrochemical
sensor. The dynamic range of detection of the sensor for the PAT gene fragment
was from 1.0 x 10(-11) to 1.0 x 10(-6) mol L(-1) and the detection limit was 1.38
x 10(-12) mol L(-1).

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17851654 [PubMed - indexed for MEDLINE]

308: Plant Physiol Biochem. 2007 Oct-Nov;45(10-11):822-33. Epub 2007 Aug 3.

Enhanced tolerance to sulfur dioxide and salt stress of transgenic Chinese
cabbage plants expressing both superoxide dismutase and catalase in chloroplasts.

Tseng MJ, Liu CW, Yiu JC.

Department of Horticulture, National Chung Hsing University, Taichung 402,
Taiwan, ROC.

To explore the possibility of overcoming the highly phytotoxic effect of SO(2)
and salt stress, we introduced the maize Cu/ZnSOD and/or CAT genes into
chloroplasts of Chinese cabbage (Brassica campestris L. ssp. pekinensis cv.
Tropical Pride) (referred to as SOD, CAT and SOD+CAT plants). SOD+CAT plants
showed enhanced tolerance to 400 ppb SO(2), and visible damage was one-sixth that
of wild-type (CK) plants. In addition, when SOD+CAT plants were exposed to a high
salt treatment of 200 mM NaCl for 4 weeks, the photosynthetic activity of the
plants decreased by only 6%, whereas that of CK plants decreased by 72%. SOD
plants had higher total APX and GR activities than CK plants. As expected, SOD
plants showed levels of protection from SO(2) and salt stress that were
moderately improved compared to CK plants. However, CAT plants showed inhibition
of APX activity and provided only limited improvements in plant stress tolerance.
Moreover, SOD+CAT plants accumulated more K(+), Ca(2+) and Mg(2+) and less Na(+)
in their leaves compared with those of CK plants. These results suggest that the
expression of SOD and CAT simultaneously is suitable for the introduction of
increased multiple stress protection.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17851086 [PubMed - indexed for MEDLINE]

309: J Econ Entomol. 2007 Aug;100(4):1416-22.

Bean alpha-amylase inhibitors in transgenic peas inhibit development of pea
weevil larvae.

de Sousa-Majer MJ, Hardie DC, Turner NC, Higgins TJ.

Department of Environmental Biology, Curtin University of Technology, P.O. Box
U1987, Perth, WA 6845, Australia. jmajer@bigpond.net.au

This glasshouse study used an improved larval measurement procedure to evaluate
the impact of transgenic pea, Pisum sativum L., seeds expressing a-amylase
inhibitor (AI)-1 or -2 proteins on pea weevil, Bruchus pisorum L. Seeds of
transgenic 'Laura' and 'Greenfeast' peas expressing alpha-(AI)-1 reduced pea
weevil survival by 93-98%. Larval mortality occurred at an early instar.
Conversely, in nontransgenic cultivars, approximately 98-99% of the pea weevils
emerged as adults. By measuring the head capsule size, we determined that larvae
died at the first to early third instar in alpha-(AI)-1 transgenic peas,
indicating that this inhibitor is highly effective in controlling this insect. By
contrast, transgenic Laura and 'Dundale' expressing alpha-(AI)-2 did not affect
pea weevil survival, but they did delay larval development. After 77 d of
development, the head capsule size indicated that the larvae were still at the
third instar stage in transgenic alpha-(AI)-2 peas, whereas adult bruchids had
developed in the nontransgenic peas.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17849896 [PubMed - indexed for MEDLINE]

310: J Econ Entomol. 2007 Aug;100(4):1129-35.

Influence of plant severing on movement of Ostrinia nubilalis larvae in Zea mays
hybrid seed production fields.

Reardon KT, Hellmich RL, Sumerford DV, Lewis LC, Reardon BJ, Calvin DD.

USDA-ARS, Corn Insects and Crop Genetics Research Unit, and Department of
Entomology, Iowa State University, 102 Genetics Laboratory, Ames, IA 50011, USA.

Genetically engineered corn hybrids that contain a cry gene from the bacterium
Bacillus thuringiensis Berliner (Bt) are gaining popularity for controlling the
corn pest Ostrinia nubilalis (Hö¼bner). Continuous use of Bt corn, however, could
select for O. nubilalis that are resistant to this corn. Monitoring for insect
resistance is important, because it could help maintain the Bt technology. A
possible monitoring method is to collect larval insects in commercial drying bins
after harvest from Bt seed production fields. A drawback to this method is that
these collections may be contaminated by insects that moved as later instars from
severed non-Bt male rows into the adjacent Bt female rows. These larvae have
little to no exposure to Bt toxin, resulting in possible "false positives." The
objectives of this study were to first find which combination of planting and
severing dates produces the least number of larvae that move from non-Bt male
plants to Bt female plants and to assess O. nubilalis larval movement from
severed non-Bt male rows to Bt female rows. Field studies in 2002 and 2003 were
designed to simulate a hybrid seed production field. Results suggest that
movement of O. nubilalis larvae from male corn is minimized when corn is planted
early and male plants are severed by 2 wk post-anthesis. This reduces the
likelihood of false positives by reducing the number of susceptible larvae moving
between Bt and non-Bt plants. Also, larvae moved to all four female rows that
were adjacent to the severed rows, but there were significantly more larvae found
in the closest row compared with the other three. These results could be used to
develop a monitoring program to find O. nubilalis larvae with resistance to Bt
corn in field populations of O. nubilalis.

Publication Types:
Evaluation Studies
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17849861 [PubMed - indexed for MEDLINE]

311: J Econ Entomol. 2007 Aug;100(4):1116-23.

Effect of age and mating status on adult European corn borer (Lepidoptera:
Crambidae) dispersal from small-grain aggregation plots.

Reardon BJ, Sappington TW.

USDA-ARS Corn Insects and Crop Genetics Research Unit, Genetics Laboratory, Iowa
State University, Ames, IA 50011, USA.

The European corn borer, Ostrinia nubilalis (Hö¼bner) (Lepidoptera: Crambidae), is
often controlled with genetically modified corn, Zea mays L., hybrids (Bacillus
thuringiensis [Bt] corn) in the United States. If Bt-resistant insects are
detected in the field, mitigation-remediation tactics must be implemented to
sustain the efficacy of insecticidal, transgenic corn. Mass releasing
laboratory-reared, susceptible adults near aggregation sites to mate with locally
emerging resistant adults is a possible remediation tactic, but it is imperative
that the former remain in or near the release site long enough to mate.
Understanding adult dispersal behavior relative to the timing of mating is
important, because it directly affects patterns of gene flow and the rate at
which Bt resistance moves through a population. Previous work shows that newly
eclosed adults do not remain in proximity to their natal field. However, moth
age, reproductive development, or mating status may influence the propensity to
disperse. The objectives of this study were to determine the effect of adult age
(0-3, 4-6, and 7-10 d old) and mating status on dispersal of adults released in
small-grain aggregation plots. Less than 1% of the marked adults released in the
aggregation plots remained after one night. More males than females were
recovered. Age influenced dispersal, with mostly 4-6-d old adults being
recovered. Conversely, mating status did not affect the number of adults
recovered. Given the paucity of marked adult moths recovered near their release
sites, mass releases of adults may not be a viable tactic to combat the spread of
resistance to Bt corn.

PMID: 17849859 [PubMed - indexed for MEDLINE]

312: Mol Plant Microbe Interact. 2007 Sep;20(9):1092-101.

The chitin-binding Cladosporium fulvum effector protein Avr4 is a virulence
factor.

van Esse HP, Bolton MD, Stergiopoulos I, de Wit PJ, Thomma BP.

Laboratory of Phytopathology, Centre for Biosystems Genomics (CBSG), Wageningen
University, Binnenhaven 5, 6709 PD Wageningen, The Netherlands.

The biotrophic fungal pathogen Cladosporium fulvum (syn. Passalora fulva) is the
causal agent of tomato leaf mold. The Avr4 protein belongs to a set of effectors
that is secreted by C. fulvum during infection and is thought to play a role in
pathogen virulence. Previous studies have shown that Avr4 binds to chitin present
in fungal cell walls and that, through this binding, Avr4 can protect these cell
walls against hydrolysis by plant chitinases. In this study, we demonstrate that
Avr4 expression in Arabidopsis results in increased virulence of several fungal
pathogens with exposed chitin in their cell walls, whereas the virulence of a
bacterium and an oomycete remained unaltered. Heterologous expression of Avr4 in
tomato increased the virulence of Fusarium oxysporum f. sp. lycopersici. Through
tomato GeneChip analyses, we demonstrate that Avr4 expression in tomato results
in the induced expression of only a few genes. Finally, we demonstrate that
silencing of the Avr4 gene in C. fulvum decreases its virulence on tomato. This
is the first report on the intrinsic function of a fungal avirulence protein that
has a counter-defensive activity required for full virulence of the pathogen.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17849712 [PubMed - indexed for MEDLINE]

313: Plant Mol Biol. 2007 Nov;65(5):645-54. Epub 2007 Sep 12.

Flavonoid profiling among wild type and related GM wheat varieties.

Ioset JR, Urbaniak B, Ndjoko-Ioset K, Wirth J, Martin F, Gruissem W, Hostettmann
K, Sautter C.

Laboratoire de Pharmacognosie et Phytochimie, Ecole Romande de Pharmacie
Genö¨ve-Lausanne, Universitö© de Genö¨ve, Quai Ernest-Ansermet 30, Geneva 4 1211,
Switzerland.

Pleiotropic effects are one of the main concerns regarding genetically modified
organisms (GMOs). This includes unintended side effects of the transgene or its
genome insertion site on the regulation of other endogenous genes, which could
potentially cause the accumulation of different secondary metabolites that may
have not only an impact on diet as repeatedly worried by the public but also on
the environment. Regarding amount and possible environmental effects, flavonoids
represent the most prominent group of secondary metabolites in wheat. Many
flavonoids function as signalling or defence molecules. We used a robust and
reproducible analytical method to compare the flavonoid content of genetically
modified (GM) wheat (Triticum aestivum L., Gramineae) expressing genes that
confer increased fungal resistance with their non-GM siblings. The transgenes
provide either a broad-spectrum fungal defence (chitinase/glucanase from barley)
or bunt-specific resistance by a viral gene (KP4). Significant differences in
flavonoid composition were found between different wheat varieties whereas
different lines of GM wheat with increased antifungal resistance showed only
minor differences in their flavonoid composition relative to their non-GM
siblings. In a field test, no significant differences were detectable between
infected and non-infected wheat of the same variety regardless of the presence of
the transgene. Our results are in agreement with the hypothesis that the
transgenes we used to increase wheat defence to fungal pathogens do not interfere
with the flavonoid biosynthesis pathway. More significantly, the genetic
background resulting from conventional breeding has a direct impact on the
biological composition of flavonoids, and thus possibly on the environment.

Publication Types:
Comparative Study

PMID: 17849220 [PubMed - indexed for MEDLINE]

314: Nat Biotechnol. 2007 Sep;25(9):981-7.

Comment in:
Nat Biotechnol. 2007 Dec;25(12):1351-4; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1354-5; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1355-6; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1355; author reply 1359-60. Nat Biotechnol. 2007 Dec;25(12):1356-8. Nat Biotechnol. 2007 Dec;25(12):1356; author reply 1359-60.

GM soybeans and health safety--a controversy reexamined.

Marshall A.

PMID: 17846624 [PubMed - indexed for MEDLINE]

315: Nat Biotechnol. 2007 Sep;25(9):950.

Comment in:
Nat Biotechnol. 2007 Nov;25(11):1213; discussion 1213.

A tragic GM "outing".

[No authors listed]

Publication Types:
Editorial

PMID: 17846608 [PubMed - indexed for MEDLINE]

316: Planta. 2008 Jan;227(2):277-86. Epub 2007 Sep 8.

Altered expression of barley proline transporter causes different growth
responses in Arabidopsis.

Ueda A, Shi W, Shimada T, Miyake H, Takabe T.

Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya,
464-8601, Japan.

A compatible solute, proline is accumulated in various kinds of plants and
microorganisms under environmental stresses. The function of proline is thought
to be an osmotic regulator under water stress, and its transport into cells is
mediated by a proline transporter. Here, we report the effects of expressing the
barley proline transporter (HvProT) under the control of either the CaMV35S
promoter (35Sp) or a root cap promoter (RCp), on Arabidopsis growth. In
Arabidopsis, transformed HvProT functions in the plasma membrane, like other
amino acid transporters. Reduction in biomass production was observed in aerial
parts of 35Sp-HvProT plants, and it was accompanied with decreased proline
accumulation in leaves. Impaired growth of 35Sp-HvProT plants was restored by
exogenously adding L: -proline. These results suggested that growth reduction was
caused by a deficiency of endogenous proline. In 35Sp-HvProT plants, the amount
of proline dehydrogenase (PDH) transcript was increased compared to wild type
(WT) plants, with a consequent enhancement of the activity of PDH. On the other
hand, the transgenic RCp-HvProT plants accumulated 2- to 3-fold more proline in
the root tip region compared to WT, and root elongation was enhanced at the same
time. Thus, different physiological responses were caused by the altered location
in accumulation of proline using two different promoters for heterologous
expression of HvProT. These results indicate the importance of proline
distribution at the tissue level during vegetative development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17828417 [PubMed - indexed for MEDLINE]

317: Planta. 2008 Jan;227(2):299-308. Epub 2007 Sep 8.

Effects of drought on water content and photosynthetic parameters in potato
plants expressing the trehalose-6-phosphate synthase gene of Saccharomyces
cerevisiae.

Stiller I, Dulai S, Kondrö¡k M, Tarnai R, Szabö¨ L, Toldi O, Bö¡nfalvi Z.

Agricultural Biotechnology Center, 2101, Gö¶dö¶llo, P.O. Box 411, Hungary.

Two transgenic potato lines, T1 and T2, expressing the trehalose-6-phosphate
synthase (TPS1) gene of yeast were isolated. In our experimental approach, we
applied two novelties, namely the fusion of the drought-inducible promoter StDS2
to TPS1 and a marker-free transformation method. In contrast to the expected
drought-induced expression, only a very low constitutive TPS1 expression was
detected in the transgenic lines, probably due to chromosomal position effects.
The observed expression pattern, however, was sufficient to alter the drought
response of plants. Detached leaves of T1 and T2 showed an 8 h delay in wilting
compared to the non-transformed control. Potted plants of T1 and T2 kept water 6
days longer than control plants and maintained high stomatal conductance and a
satisfactory rate of net photosynthesis. During drought treatment, CO2
assimilation rate measured at saturating CO2 level was maintained at maximum
level for 6-9 days in transgenic plants while it decreased rapidly after 3 days
in the wild type plants. Under optimal growth conditions, lower CO2 fixation was
detected in the transgenic than in the control plants. Stomatal densities of T1
and T2 leaves were reduced by 30-40%. This may have contributed to the lower CO2
fixation rate and altered drought response.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17828416 [PubMed - indexed for MEDLINE]

318: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 27;363(1492):761-76.

Biological control and sustainable food production.

Bale JS, van Lenteren JC, Bigler F.

School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT,
UK. j.s.bale@bham.ac.uk

The use of biological control for the management of pest insects pre-dates the
modern pesticide era. The first major successes in biological control occurred
with exotic pests controlled by natural enemy species collected from the country
or area of origin of the pest (classical control). Augmentative control has been
successfully applied against a range of open-field and greenhouse pests, and
conservation biological control schemes have been developed with indigenous
predators and parasitoids. The cost-benefit ratio for classical biological
control is highly favourable (1:250) and for augmentative control is similar to
that of insecticides (1:2-1:5), with much lower development costs. Over the past
120 years, more than 5000 introductions of approximately 2000 non-native control
agents have been made against arthropod pests in 196 countries or islands with
remarkably few environmental problems. Biological control is a key component of a
'systems approach' to integrated pest management, to counteract
insecticide-resistant pests, withdrawal of chemicals and minimize the usage of
pesticides. Current studies indicate that genetically modified insect-resistant
Bt crops may have no adverse effects on the activity or function of predators or
parasitoids used in biological control. The introduction of rational approaches
for the environmental risk assessment of non-native control agents is an
essential step in the wider application of biological control, but future success
is strongly dependent on a greater level of investment in research and
development by governments and related organizations that are committed to a
reduced reliance on chemical control.

Publication Types:
Review

PMID: 17827110 [PubMed - indexed for MEDLINE]

319: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 27;363(1492):741-59.

Sustainable agriculture and plant diseases: an epidemiological perspective.

Gilligan CA.

Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge
CB2 3EA, UK. cag1@cam.ac.uk

The potential for modern biology to identify new sources for genetical, chemical
and biological control of plant disease is remarkably high. Successful
implementation of these methods within globally and locally changing agricultural
environments demands new approaches to durable control. This, in turn, requires
fusion of population genetics and epidemiology at a range of scales from the
field to the landscape and even to continental deployment of control measures. It
also requires an understanding of economic and social constraints that influence
the deployment of control. Here I propose an epidemiological framework to model
invasion, persistence and variability of epidemics that encompasses a wide range
of scales and topologies through which disease spreads. By considering how to map
control methods onto epidemiological parameters and variables, some new
approaches towards optimizing the efficiency of control at the landscape scale
are introduced. Epidemiological strategies to minimize the risks of failure of
chemical and genetical control are presented and some consequences of
heterogeneous selection pressures in time and space on the persistence and
evolutionary changes of the pathogen population are discussed. Finally, some
approaches towards embedding epidemiological models for the deployment of control
in an economically plausible framework are presented.

Publication Types:
Review

PMID: 17827101 [PubMed - indexed for MEDLINE]

320: J Agric Food Chem. 2007 Oct 3;55(20):8003-10. Epub 2007 Sep 8.

A strategy for designing multi-taxa specific reference gene systems. example of
application--ppi phosphofructokinase (ppi-PPF) used for the detection and
quantification of three taxa: maize (Zea mays), cotton (Gossypium hirsutum) and
rice (Oryza sativa).

Chaouachi M, Giancola S, Romaniuk M, Laval V, Bertheau Y, Brunel D.

Unitö© Etude du Polymorphisme des Gö©nomes Vö©gö©taux (EPGV), Institut National de la
Recherche Agronomique (INRA), Centre National de Gö©notypage (CNG), 2 Rue Gaston
Crö©mieux 91057, CP5721, Evry Cedex, France.

In the first part of the paper, we report the description of a new strategy for
the development of a plant reference gene system that can be used for genetically
modified organism (GMO) analysis. On the basis of in silico research for
candidate genes, the design of degenerate primers allowed the obtention of
genomic sequences of the selected gene ppi-phosphofructokinase ( ppi-PPF) for
nine taxa in which GMOs have been developed. The comparison and the analysis of
inter- and intraspecies sequence variability were performed using a large number
of species and cultivars. As an example of application following the detection of
single nucleotide polymorphism, we designed specific conventional and real-time
polymerase chain reaction tests for the detection and quantification of three
taxa, namely, maize, cotton, and rice. This system was highly specific and
sensitive. The gene copy number conservation among different cultivars was
analyzed and confirmed with a sequencing step. This reference gene system is
adequate for use in routine assays for the quantification of GMOs. We then
explain briefly the constraints faced and propose recommendations when designing
a reference gene system depending on the species to be targeted.

PMID: 17824661 [PubMed - indexed for MEDLINE]

321: New Phytol. 2007;176(3):665-72. Epub 2007 Aug 23.

Physiological and genetic mapping study of tolerance to root-knot nematode in
rice.

Shrestha R, Uzzo F, Wilson MJ, Price AH.

School of Biological Sciences, University of Aberdeen, Aberdeen, AB24 3UU, UK.
r.shrestha@abdn.ac.uk

The root-knot nematode Meloidogyne graminicola is an obligate biotrophic parasite
and a major pest of rice (Oryza sativa) for which resistant varieties are not
currently available. Quantitative trait loci (QTLs) for partial resistance to M.
graminicola were identified using a mapping population based on two rice
varieties, Bala x Azucena. Experiments were carried out to investigate the
interactions between M. graminicola and these two varieties in terms of nematode
establishment, reproduction and effect on rice yield. Nematode establishment was
also assessed in the mapping population. Meloidogyne graminicola consistently
caused more galling and had higher reproductive success in Azucena than in Bala.
M. graminicola did not significantly reduce yield in Bala, but caused a yield
reduction of almost half in Azucena, suggesting that the partial resistance to
nematode establishment was related to nematode tolerance. A total of six
significant or putative QTLs for nematode tolerance were detected. For two of the
QTLs detected, Azucena was the donor of the tolerance alleles, suggesting it may
be possible to breed plants with greater tolerance than Bala.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17822410 [PubMed - indexed for MEDLINE]

322: Mol Genet Genomics. 2007 Dec;278(6):709-22. Epub 2007 Sep 6.

Characterization of two rice peroxidase promoters that respond to blast
fungus-infection.

Sasaki K, Yuichi O, Hiraga S, Gotoh Y, Seo S, Mitsuhara I, Ito H, Matsui H,
Ohashi Y.

Division of Plant Sciences, National Institute of Agrobiological Sciences,
Tsukuba, Ibaraki 305-8602, Japan.

Peroxidase (POX) genes consist of a large gene family possibly contributing to
self-defense, however constitutive and stress-induced expression patterns of
individual gene were poorly understood in rice. We studied here the
characteristic expression of two representative rice POX genes, R2329 and R2184,
which are blast fungus-inducible (Sasaki et al. in Plant Cell Physiol
45:1442-1452, 2004). Basal GUS activity in R2329 promoter::GUS rice plants was
100-fold higher than that in R2184 promoter::GUS plants, and these levels
reflected the transcript levels monitored by quantitative real-time RT-PCR. R2329
promoter was activated by blast fungus-infection and wounding, and R2184 promoter
was activated by the fungal-infection and methyl jasmonate (MeJA)-treatment. By
histochemical GUS staining analysis, constitutive R2329 and R2184 expression was
commonly found in vascular bundle and exodermis in leaves and roots, while the
precise expression profile was characteristic. In blast fungus inoculated R2329
promoter::GUS leaves, GUS staining was induced just around fungus-induced local
lesions. Analysis of the 5' deleted promoters suggests the presence of many kinds
of stress-responsive elements in the regions between -1798 and -748 of R2329
promoter and between -1975 and -548 of R2184 promoter. These results revealed the
stress-responsive characteristics of R2329 and R2184 promoters, and indicated the
possible use for generation of useful transgenic plants.

PMID: 17805575 [PubMed - indexed for MEDLINE]

323: Nature. 2007 Sep 6;449(7158):9.

Biotech crop rules get rewrite.

Marris E.

Publication Types:
News

PMID: 17805261 [PubMed - indexed for MEDLINE]

324: Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14867-71. Epub 2007 Sep 5.

The small interfering RNA production pathway is required for shoot meristem
initiation in rice.

Nagasaki H, Itoh J, Hayashi K, Hibara K, Satoh-Nagasawa N, Nosaka M, Mukouhata M,
Ashikari M, Kitano H, Matsuoka M, Nagato Y, Sato Y.

Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya
464-8601, Japan.

The shoot apical meristem (SAM) is a group of stem cells that are responsible for
plant development. Mutations in rice SHOOTLESS2 (SHL2), SHL4/SHOOT ORGANIZATION2
(SHO2), and SHO1 cause complete deletion or abnormal formation of the SAM. In
this study we showed that defects in SAM formation in shl mutants are associated
with the loss of expression of the homeodomain-leucine zipper (HD-ZIPIII) family
genes. Rice SHL2, SHL4/SHO2, and SHO1 encoded orthologues of Arabidopsis
RNA-dependent RNA polymerase 6, ARGONAUTE (AGO) 7, and DICER-like 4,
respectively, whose mutations affect leaf development through the trans-acting
siRNA (ta-siRNA) pathway. This suggested that the ta-siRNA pathway regulates the
critical step of SAM formation during rice embryogenesis. The gain-of-function
experiment by the ectopic expression of SHL4 resulted in reduced accumulation of
an microRNA, miR166, and partial adaxialization of leaves, supporting a role for
the ta-siRNA pathway in the maintenance of leaf polarity as previously reported
in maize. Analysis of the spatiotemporal expression patterns of HD-ZIPIII and
miR166 in wild-type and shl mutant embryos suggested that the loss of HD-ZIPIII
expression in the SAM region of the developing embryo is the result of ectopic
expression of miR166. Our analysis of shl mutants demonstrated that HD-ZIPIII
expression regulated by miR166 is sensitive to the ta-siRNA pathway during SAM
formation in rice embryogenesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17804793 [PubMed - indexed for MEDLINE]

325: Plant Cell Physiol. 2007 Oct;48(10):1472-83. Epub 2007 Sep 5.

Temperature response of photosynthesis in transgenic rice transformed with
'sense' or 'antisense' rbcS.

Makino A, Sage RF.

Graduate School of Agricultural Science, Tohoku University, 1-1
Tsutsumidori-Amamiyamachi, Sendai, 981-8555 Japan. makino@biochem.tohoku.ac.jp

The responses of chlorophyll fluorescence, gas exchange rate and Rubisco
activation state to temperature were examined in transgenic rice plants with 130
and 35% of the wild-type (WT) Rubisco content by transformation with rbcS cDNA in
sense and antisense orientations, respectively. Although the optimal temperatures
of PSII quantum efficiency and CO(2) assimilation were found to be between 25 and
32 degrees C, the maximal activation state of Rubisco was found to be between 16
and 20 degrees C in all genotypes. The Rubisco flux control coefficient was also
the highest between 16 and 20 degrees C in the WT and antisense lines [>0.88 at
an intercellular CO(2) pressure (Ci) of 28 Pa]. Gross photosynthesis at Ci = 28
Pa per Rubisco content in the WT between 12 and 20 degrees C was close to that of
the antisense lines where high Rubisco control is present. Thus, Rubisco activity
most strongly limited photosynthesis at cool temperatures. These results
indicated that a selective enhancement of Rubisco content can enhance
photosynthesis at cool temperatures, but in the sense line with enhanced Rubisco
content Pi regeneration limitation occurred. Above 20 degrees C, the Rubisco flux
control coefficient declined. This decline was associated with a decline in
Rubisco activation. The activation state of Rubisco measured at each temperature
decreased with increasing Rubisco content, and the slope of activation to Rubisco
content was independent of temperature. We discuss the possibility that the
decline in Rubisco activation at intermediate and high temperatures is part of a
regulated response to a limitation in other photosynthetic processes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17804480 [PubMed - indexed for MEDLINE]

326: Endocr Rev. 2007 Oct;28(6):664-84. Epub 2007 Sep 4.

Neuropeptide y receptor selective ligands in the treatment of obesity.

Kamiji MM, Inui A.

Department of Gastroenterology, Faculty of Medicine, University of Sao Paulo,
Ribeirö¸o Preto Campus 14048-900, Ribeirö¸o Preto-SP, Brazil.

Obesity is a serious public health problem throughout the world, affecting both
developed societies and developing countries. The central nervous system has
developed a meticulously interconnected circuitry in order to keep us fed and in
an adequate nutritional state. One of these consequences is that an energy-dense
environment favors the development of obesity. Neuropeptide Y (NPY) is one of the
most abundant and widely distributed peptides in the central nervous system of
both rodents and humans and has been implicated in a variety of physiological
actions. Within the hypothalamus, NPY plays an essential role in the control of
food intake and body weight. Centrally administered NPY causes robust increases
in food intake and body weight and, with chronic administration, can eventually
produce obesity. NPY activates a population of at least six G protein-coupled Y
receptors. NPY analogs exhibit varying degrees of affinity and specificity for
these Y receptors. There has been renewed speculation that ligands for Y
receptors may be of benefit for the treatment of obesity. This review highlights
the therapeutic potential of Y(1), Y(2), Y(4), and Y(5) receptor agonists and
antagonists as additional intervention to treat human obesity.

Publication Types:
Review

PMID: 17785427 [PubMed - indexed for MEDLINE]

327: Sci Am. 2007 Sep;297(3):104-11.

Sowing a gene revolution.

Raney T, Pingali P.

Agricultural and Development Economics Division, United Nations Food and
Agriculture Organization, Rome.

PMID: 17784631 [PubMed - indexed for MEDLINE]

328: Sci Am. 2007 Sep;297(3):54-6, 58.

A question of sustenance.

Stix G.

PMID: 17784624 [PubMed - indexed for MEDLINE]

329: J AOAC Int. 2007 Jul-Aug;90(4):1098-106.

Characterization of genetically modified maize in weakly contaminated seed
batches and identification of the origin of the adventitious contamination.

Petit L, Pagny G, Baraige F, Nignol AC, Zhang D, Fach P.

Agence Franö§aise de Sö©curitö© Sanitaire des Aliments, Laboratoire d'Etudes et de
Recherches sur la Qualitö© des Aliments et sur les Procö©dö©s Agro-Alimentaires,
Unitö© EBA, 23 Avenue du Gö©nö©ral De Gaulle, 94700 Maisons-Alfort, France.

So far, relatively few genetically modified plants (GMPs) have been planted in
the European Union (EU). However, in France, seed batches weakly contaminated by
unidentified GM materials have recently been detected among commercial maize
seeds (14 seed batches positive out of 447 analyzed). We have developed a 3-step
approach to precisely identify the genetic modifications detected in such maize
seed batches. First, to isolate GMPs derived from the contaminated seed batches,
10 000 maize seeds of each batch were planted and screened by polymerase chain
reaction (PCR) on 100-plant batches, then on 10-plant subbatches, and finally,
plant by plant. In a second step, specific identification of the individual GMPs
was performed. Finally, to determine the origin of the contamination, each
individual GMP was analyzed by simple sequence repeat (SSR) markers. The results
showed that all batches were contaminated by few GM seeds, having a GM content <
0.1%. Finally, 12 individual GMPs have been isolated from 17 plant pools that
were tested positive either for P35-S and/or T-Nos. MON810 and T25 transformation
events approved for cultivation in the EU were detected in 7 individual GMPs. The
other seed batches were contaminated by genetically modified organisms (GMOs)
that are not approved in the EU, including GA21 or the stacking MON810/T25.
Presumable identification of T14 was also achieved following sequencing of 1
individual GMP. The data also showed that most of the seed batches were
contaminated by several transformation events. Finally, analysis of SSR markers
indicated that the contaminations were essentially due to cross-pollination in
the seed production process.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17784498 [PubMed - indexed for MEDLINE]

330: Plant Physiol. 2007 Oct;145(2):351-66. Epub 2007 Aug 31.

Expression of aberrant forms of AUXIN RESPONSE FACTOR8 stimulates parthenocarpy
in Arabidopsis and tomato.

Goetz M, Hooper LC, Johnson SD, Rodrigues JC, Vivian-Smith A, Koltunow AM.

Commonwealth Scientific and Industrial Research Organization, Plant Industry,
Glen Osmond, South Australia 5064, Australia.

Fruit initiation in Arabidopsis (Arabidopsis thaliana) is generally repressed
until fertilization occurs. However, mutations in AUXIN RESPONSE FACTOR8 (ARF8)
uncouple fruit initiation from fertilization, resulting in the formation of
seedless, parthenocarpic fruit. Here we induced parthenocarpy in wild-type
Arabidopsis by introducing either the mutant genomic (g) Atarf8-4 sequence or
gAtARF8:beta-glucuronidase translational fusion constructs by plant
transformation. Silencing of endogenous AtARF8 transcription was not observed,
indicating that the introduced, aberrant ARF8 transcripts were compromising the
function of endogenous ARF8 and/or associated factors involved in suppressing
fruit initiation. To analyze the role of ARF8 in tomato (Solanum lycopersicum) we
initially emasculated 23 tomato cultivars to test for background parthenocarpy.
Surprisingly, all had a predisposition to initiate fertilization-independent
fruit growth. Expression of gAtarf8-4 in transgenic tomato ('Monalbo') resulted
in a significant increase in the number and size of parthenocarpic fruit.
Isolation of tomato ARF8 cDNA indicated significant sequence conservation with
AtARF8. SlARF8 may therefore control tomato fruit initiation in a similar manner
as AtARF8 does in Arabidopsis. Two SlARF8 cDNAs differing in size by 5 bp were
found, both arising from the same gene. The smaller cDNA is a splice variant and
is also present in Arabidopsis. We propose that low endogenous levels of the
splice variant products might interfere with efficient formation/function of a
complex repressing fruit initiation, thereby providing an explanation for the
observed ovary expansion in tomato and also Arabidopsis after emasculation.
Increasing the levels of aberrant Atarf8-4 transcripts may further destabilize
formation/function of the complex in a dosage-dependent manner enhancing tomato
parthenocarpic fruit initiation frequency and size and mimicking the
parthenocarpic dehiscent silique phenotype found in homozygous Atarf8-4 mutants.
Collectively these data suggest that similar mechanisms involving auxin signaling
exist to inhibit parthenocarpic fruit set in tomato and Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17766399 [PubMed - indexed for MEDLINE]

331: Plant Biotechnol J. 2007 Nov;5(6):835-46. Epub 2007 Aug 31.

superwoman1-cleistogamy, a hopeful allele for gene containment in GM rice.

Yoshida H, Itoh J, Ohmori S, Miyoshi K, Horigome A, Uchida E, Kimizu M, Matsumura
Y, Kusaba M, Satoh H, Nagato Y.

Hokuriku Research Center, National Agricultural Research Center, Niigata
943-0193, Japan. yocida@affrc.go.jp

Cleistogamy is an efficient strategy for preventing gene flow from genetically
modified (GM) crops. We identified a cleistogamous mutant of rice harbouring a
missense mutation (the 45th residue isoleucine to threonine; I45T) in the class-B
MADS-box gene SUPERWOMAN1 (SPW1), which specifies the identities of lodicules
(equivalent to petals) and stamens. In the mutant, spw1-cls, the stamens are
normal, but the lodicules are transformed homeotically to lodicule-glume mosaic
organs, thereby engendering cleistogamy. Since this mutation does not affect
other agronomic traits, it can be used in crosses to produce transgenic lines
that do not cause environmental perturbation. Molecular analysis revealed that
the reduced heterodimerization ability of SPW1(I45T) with its counterpart class-B
proteins OsMADS2 and OsMADS4 caused altered lodicule identity. spw1-cls is the
first useful mutant for practical gene containment in GM rice. Cleistogamy is
possible in many cereals by engineering class-B floral homeotic genes and thereby
inducing lodicule identity changes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17764519 [PubMed - indexed for MEDLINE]

332: Plant J. 2007 Oct;52(2):362-73. Epub 2007 Aug 30.

StGA2ox1 is induced prior to stolon swelling and controls GA levels during potato
tuber development.

Kloosterman B, Navarro C, Bijsterbosch G, Lange T, Prat S, Visser RG, Bachem CW.

Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding,
Department of Plant Sciences, Wageningen University and Research Centre, PO Box
386, 6700 AJ Wageningen, The Netherlands. bjorn.kloosterman@wur.nl

The formation and growth of a potato (Solanum tuberosum) tuber is a complex
process regulated by different environmental signals and plant hormones. In
particular, the action of gibberellins (GAs) has been implicated in different
aspects of potato tuber formation. Here we report on the isolation and functional
analysis of a potato GA 2-oxidase gene (StGA2ox1) and its role in tuber
formation. StGA2ox1 is upregulated during the early stages of potato tuber
development prior to visible swelling and is predominantly expressed in the
subapical region of the stolon and growing tuber. 35S-over-expression
transformants exhibit a dwarf phenotype, reduced stolon growth and earlier in
vitro tuberization. Transgenic plants with reduced expression levels of StGA2ox1
showed normal plant growth, an altered stolon swelling phenotype and delayed in
vitro tuberization. Tubers of the StGA2ox1 suppression clones contain increased
levels of GA20, indicating altered GA metabolism. We propose a role for StGA2ox1
in early tuber initiation by modifying GA levels in the subapical stolon region
at the onset of tuberization, thereby facilitating normal tuber development and
growth.

PMID: 17764503 [PubMed - indexed for MEDLINE]

333: Planta. 2007 Dec;227(1):263-72. Epub 2007 Sep 1.

Characterization of rice tryptophan decarboxylases and their direct involvement
in serotonin biosynthesis in transgenic rice.

Kang S, Kang K, Lee K, Back K.

Department of Molecular Biotechnology, Agricultural Plant Stress Research Center,
Biotechnology Research Institute, Chonnam National University, Gwangju, South
Korea.

L-Tryptophan decarboxylase (TDC) and L-tyrosine decarboxylase (TYDC) belong to a
family of aromatic L-amino acid decarboxylases and catalyze the conversion of
tryptophan and tyrosine into tryptamine and tyramine, respectively. The rice
genome has been shown to contain seven TDC or TYDC-like genes. Three of these
genes for which cDNA clones were available were characterized to assign their
functions using heterologous expression in Escherichia coli and rice (Oryza
sativa cv. Dongjin). The purified products of two of the genes were expressed in
E. coli and exhibited TDC activity, whereas the remaining gene could not be
expressed in E. coli. The recombinant TDC protein with the greatest TDC activity
showed a K (m) of 0.69 mM for tryptophan, and its activity was not inhibited by
phenylalanine or tyrosine, indicating a high level of substrate specificity
toward tryptophan. The ectopic expression of the three cDNA clones in rice led to
the abundant production of the products of the encoded enzymes, tyramine and
tryptamine. The overproduction of TYDC resulted in stunted growth and a lack of
seed production due to tyramine accumulation, which increased as the plant aged.
In contrast, transgenic plants that produced TDC showed a normal phenotype and
contained 25-fold and 11-fold higher serotonin in the leaves and seeds,
respectively, than the wild-type plants. The overproduction of either tyramine or
serotonin was not strongly related to the enhanced synthesis of tyramine or
serotonin derivatives, such as feruloyltyramine and feruloylserotonin, which are
secondary metabolites that act as phytoalexins in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17763868 [PubMed - indexed for MEDLINE]

334: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 27;363(1492):905-13.

The role of biotechnology for agricultural sustainability in Africa.

Thomson JA.

Department of Molecular and Cell Biology, University of Cape Town, Cape Town
7701, Republic of South Africa. jennifer.thomson@uct.ac.za

Sub-Saharan Africa could have a shortfall of nearly 90Mt of cereals by the year
2025 if current agricultural practices are maintained. Biotechnology is one of
the ways to improve agricultural production. Insect-resistant varieties of maize
and cotton suitable for the subcontinent have been identified as already having a
significant impact. Virus-resistant crops are under development. These include
maize resistant to the African endemic maize streak virus and cassava resistant
to African cassava mosaic virus. Parasitic weeds such as Striga attack the roots
of crops such as maize, millet, sorghum and upland rice. Field trials in Kenya
using a variety of maize resistant to a herbicide have proven very successful.
Drought-tolerant crops are also under development as are improved varieties of
local African crops such as bananas, cassava, sorghum and sweet potatoes.

Publication Types:
Review

PMID: 17761472 [PubMed - indexed for MEDLINE]

335: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 27;363(1492):703-16.

Breeding for abiotic stresses for sustainable agriculture.

Witcombe JR, Hollington PA, Howarth CJ, Reader S, Steele KA.

CAZS Natural Resources, University of Wales, Bangor LL57 2UW, UK.
j.r.witcombe@bangor.ac.uk

Using cereal crops as examples, we review the breeding for tolerance to the
abiotic stresses of low nitrogen, drought, salinity and aluminium toxicity. All
are already important abiotic stress factors that cause large and widespread
yield reductions. Drought will increase in importance with climate change, the
area of irrigated land that is salinized continues to increase, and the cost of
inorganic N is set to rise. There is good potential for directly breeding for
adaptation to low N while retaining an ability to respond to high N conditions.
Breeding for drought and salinity tolerance have proven to be difficult, and the
complex mechanisms of tolerance are reviewed. Marker-assisted selection for
component traits of drought in rice and pearl millet and salinity tolerance in
wheat has produced some positive results and the pyramiding of stable
quantitative trait locuses controlling component traits may provide a solution.
New genomic technologies promise to make progress for breeding tolerance to these
two stresses through a more fundamental understanding of underlying processes and
identification of the genes responsible. In wheat, there is a great potential of
breeding genetic resistance for salinity and aluminium tolerance through the
contributions of wild relatives.

Publication Types:
Review

PMID: 17761467 [PubMed - indexed for MEDLINE]

336: Plant Physiol Biochem. 2007 Sep;45(9):722-8. Epub 2007 Jul 25.

Enhanced tolerance to heat stress in transgenic plants expressing the GASA4 gene.

Ko CB, Woo YM, Lee DJ, Lee MC, Kim CS.

Department of Plant Biotechnology and Agricultural Plant Stress Research Center,
Chonnam National University, 300 Yongbong-dong, Buk-gu, Kwangju 500-757, South
Korea.

We conducted a genetic yeast screen to identify Thermo-tolerance genes (TTOs) in
maize kernel cDNA library. During the screening, we identified a maize clone
(TTO6) that seemed to confer elevated heat tolerance in comparison to control
cells. TTO6 cDNA (GenBank accession no. AY103785) encodes an 11-kDa protein which
is 69% similarity to the Arabidopsis GASA4 gene. To further examine heat
tolerance in Arabidopsis, we functionally characterized the GASA4 gene and found
that heat induced GASA4 expression. Constitutive expression of GASA4 in
Arabidopsis led to elevated heat tolerance in transgenic lines. Interestingly,
endoplasmic reticulum chaperone expression analysis suggests that GASA4
influences BiP gene expression during heat stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17761429 [PubMed - indexed for MEDLINE]

337: Plant Physiol Biochem. 2007 Sep;45(9):637-46. Epub 2007 Jul 25.

Production and phenotypic analysis of rice transgenics with altered levels of
pyruvate decarboxylase and alcohol dehydrogenase proteins.

Agarwal S, Kapoor A, Lakshmi OS, Grover A.

Department of Plant Molecular Biology, University of Delhi, South Campus, Benito
Juarez Road, Dhaula Kuan, New Delhi 110021, India.

Pyruvate decarboxylase (Pdc) and alcohol dehydrogenase (Adh) enzymes are
responsible for the operation of ethanolic fermentation pathway that appears to
correlate to an extent with anoxia tolerance in plants. This study was undertaken
with the objective of (a) analysing the rice pdc gene family and (b) altering the
efficacy of the ethanolic fermentation process, through production of transgenic
rice plants over- and under-expressing pyruvate decarboxylase (employing Ospdc1
gene from rice) as well as over-expressing alcohol dehydrogenase (employing
Ghadh2 gene from cotton) proteins. Correlations noted in this study between the
pattern of expression of the Pdc alpha-subunit and Ospdc2 transcript as well as
between the Pdc beta-subunit and Ospdc1 transcript suggest the possibility that
alpha-subunit is encoded by Ospdc2 and that beta-subunit is encoded by Ospdc1.
The fact that levels of Pdc beta-subunit were particularly high in pUH-sPdc1
(plasmid construct designed for over-expression of Ospdc1) seedlings while levels
of beta-subunit levels were negligible or lower in pUH-asPdc1 (plasmid construct
designed for under-expression of Ospdc1) seedlings also support these
observations. Transgenics raised for over-expression of Pdc and Adh and
under-expression of Pdc were confirmed for the transgene presence and effects by
PCR, Southern blotting, Northern blotting, Western blotting and isozyme assays.
Pdc and Adh over-expressing rice transgenics at early seedling stage under
unstressed control growth conditions showed slight, consistent advantage in root
vigour as compared to that of wild-type seedlings.

PMID: 17761427 [PubMed - indexed for MEDLINE]

338: Arch Anim Nutr. 2007 Aug;61(4):308-16.

Nutritional assessment of genetically modified rapeseed synthesizing high amounts
of mid-chain fatty acids including production responses of growing-finishing
pigs.

Bö¶hme H, Rudloff E, Schö¶ne F, Schumann W, Hö¼ther L, Flachowsky G.

Institute of Animal Nutrition, Federal Agricultural Research Centre (FAL),
Braunschweig, Germany.

The nutritive value of genetically modified myristic acid-rich rapeseed, in which
a acyl-thioesterase gene inserted, was studied. Crude nutrients, amino acid and
fatty acid profiles as well as mineral and glucosinolate contents were determined
and compared with those of the non-transgenic parental cultivar. The
concentration of crude nutrients, minerals and amino acids were found to be
within the range of natural variance. The myristic and palmitic acid content
increased from 0.1 - 11.4% and from 3.6-20%, respectively, at the expense of
oleic acid, which decreased from 68.6-42.6% of total fatty acids. The
glucosinolate contents increased from 12.4 micromol/g in the parental plant to 19
micromol/g DM in the GM-plant. Full-fat rapeseed of both cultivars was
incorporated in pig diets at a level of 15%, and the digestibility and the
production efficiency were tested under ad libitum feeding conditions with ten
pigs each over the growing finishing period from 32-105 kg BW. The experimental
diets did not show significant differences in digestibility and energetic feeding
value. However, feed intake and weight gain decreased presumably due to the
increasing glucosinolate intake associated with the feeding of transgenic
rapeseed. The dietary fatty acids profile influenced the fatty acid profile of
body fat. Myristic acid accumulated in back fat and intramuscular fat while the
oleic acid content decreased. The increased glucosinolate intake affected the
weight of thyroid glands and their iodine concentration.

PMID: 17760308 [PubMed - indexed for MEDLINE]

339: J Exp Bot. 2007;58(12):3183-95. Epub 2007 Aug 28.

Uptake and allocation of carbon and nitrogen in Vicia narbonensis plants with
increased seed sink strength achieved by seed-specific expression of an amino
acid permease.

Gö¶tz KP, Staroske N, Radchuk R, Emery RJ, Wutzke KD, Herzog H, Weber H.

Fachgebiet Pflanzenbau in den Tropen und Subtropen, Humboldt Universitö¤t zu
Berlin, D-14195 Berlin, Germany.

Over-expressing an amino acid permease in Vicia narbonensis seeds increases sink
strength for N that is evident from the higher seed protein content and seed
weight. Here, the effect of increased seed sink strength of line AAP-12 on
growth, development, and on whole plant carbon and nitrogen uptake and
partitioning is analysed. AAP-12 plants have a prolonged growth period.
Accumulation and partitioning of dry matter and N in leaves, stems, and pods are
higher whereas remobilization to the seeds is delayed, indicating that the switch
from growth to reserve allocation and remobilization is delayed. Measuring uptake
and allocation of (15)N-ammonia applied via the roots revealed a higher and
longer label uptake period during maturation. Measuring whole plant carbon
fixation and allocation after (13)C labelling shows higher levels at maturation,
particularly in seeds, indicating higher seed sink strength for C and increased
allocation into maturing seeds. Levels of cytokinins were dramatically increased
in AAP-12 seeds indicating its role in nitrogen-mediated growth stimulation.
AAP-12 seeds have higher natural abundances for (13)C indicating increased C
fixation via PEP carboxylase in order to meet the higher demand of carbon
acceptors for amino acid synthesis. In summary, increased seed sink strength for
N in AAP-12 stimulates seed growth, but also that of vegetative organs, which
finally leads to a higher ratio of vegetative to seed biomass at maturity and
thus a lower harvest index. Therefore, the increased N uptake due to higher seed
demand of AAP-12 is partly compensated by growth stimulation of vegetative
organs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17728294 [PubMed - indexed for MEDLINE]

340: Int J Food Microbiol. 2007 Oct 20;119(1-2):147-51. Epub 2007 Jul 31.

Opportunities for biotechnology and policy regarding mycotoxin issues in
international trade.

Kendra DF, Dyer RB.

USDA-Agricultural Research Service, National Center for Agricultural Utilization
Research, 1815 N. University Street, Peoria, Illinois 61604, USA.
david.kendra@ars.usda.gov

Despite being introduced more than a decade ago, agricultural biotechnology still
remains framed in controversy impacting both the global economy and international
regulations. Controversies surrounding agricultural biotechnology produced crops
and foods commonly focus on human and environmental safety, intellectual property
rights, consumer choice, ethics, food security, poverty reduction and
environmental conservation. Originally, some consumers were reluctant to accept
the first generation agricultural biotechnology products because they appeared to
primarily benefit agricultural producers; however, it is clear from continued
evaluations that these technologies also improved both the safety and
wholesomeness of food and helped improve the environment. Plants engineered to
resist insect pests and tolerate less toxic pesticides resulted in improved
yields thereby enabling farmers to produce more food per acre while reducing the
need for herbicides, pesticides, and water and tilling. An indirect benefit of
reduced pest damage in transgenic corn expressing genes to control insect pests
is lower levels of mycotoxins, most notably those caused by the genus Fusarium.
Mycotoxins are an important regulatory issue globally because of their toxic and
carcinogenic potential to humans and animals. Complicating this issue is the fact
that toxicological databases for mycotoxins are relatively incomplete compared to
other food contaminants. Current debates about agricultural biotechnology and
mycotoxins reveal significant differences in perception of associated risks and
benefits. When faced with uncertainty, regulators tend to set limits as low as
possible. Additionally, some regulators invoke the "Precautionary Principle" when
limited information is available or disputes over interpretation exist for
possible contaminants, including mycotoxins. A major concern regarding use of the
"Precautionary Principle" is the appearance that regulators can justify setting
any limit on the basis of inconclusive or unknown potential hazards of a
contaminant which may significantly impact global trade because mycotoxin
residues vary widely between countries. This paper describes the current economic
and heath impact of these regulations and their impact on international trade.

Publication Types:
Review

PMID: 17727996 [PubMed - indexed for MEDLINE]

341: Ir Med J. 2007 May;100(5):475-6.

Genetically modified food and health--a cause for concern?

Cullen E.

Publication Types:
Letter

PMID: 17727126 [PubMed - indexed for MEDLINE]

342: Plant Biotechnol J. 2008 Jan;6(1):13-21. Epub 2007 Aug 28.

Modifying lysine biosynthesis and catabolism in corn with a single bifunctional
expression/silencing transgene cassette.

Frizzi A, Huang S, Gilbertson LA, Armstrong TA, Luethy MH, Malvar TM.

Mystic Research, Monsanto Company, 62 Maritime Drive, Mystic, CT 06355, USA.

Although it is one of the major crops in the world, corn has poor nutritional
quality for human and animal consumption due to its low lysine content. Here, we
report a method of simultaneous expression of a deregulated lysine biosynthetic
enzyme, CordapA, and reduction of a bifunctional lysine degradation enzyme,
lysine-ketoglutarate reductase/saccharophine dehydrogenase (LKR/SDH), in
transgenic corn plants by a single transgene cassette. This is accomplished by
inserting an inverted-repeat sequence targeting the maize LKR/SDH gene into an
intron of a transgene cassette that expresses CordapA. This combination of
LKR/SDH silencing and CordapA expression led to the accumulation of free lysine
to over 4000 p.p.m. in transgenic corn grain, compared to less than 100 p.p.m. in
wild-type controls. This intron-embedded silencing cassette design reduces the
number of transgene cassettes needed in transgenic approaches for manipulating
metabolic pathways that sometimes require expression of one gene and silencing of
another.

PMID: 17725550 [PubMed - indexed for MEDLINE]

343: Invest Ophthalmol Vis Sci. 2007 Sep;48(9):4226-31.

The prime role of HDL to transport lutein into the retina: evidence from
HDL-deficient WHAM chicks having a mutant ABCA1 transporter.

Connor WE, Duell PB, Kean R, Wang Y.

Division of Endocrinology, Metabolism, and Clinical Nutrition, Department of
Medicine, Oregon Health and Science University, Portland, Oregon 97239, USA.
connorw@ohsu.edu

PURPOSE: Lutein and zeaxanthin are largely transported in plasma by high-density
lipoprotein (HDL). The Wisconsin hypoalpha mutant (WHAM) chicken has a recessive
sex-linked mutation in the ABCA1 transporter gene that results in a severe
deficiency of HDL. In this study, the transport and tissue distribution of lutein
and zeaxanthin were examined in newly hatched and 28-day-old WHAM chicks compared
with control chicks. METHODS: One-day-old WHAM and control chicks were randomized
to be fed a high-lutein or a control diet for 28 days. The plasma and tissues
were analyzed for lutein, zeaxanthin, and lipoproteins on days 1 and 28. RESULTS:
The WHAM chicks had very low plasma levels of HDL cholesterol (5.3% of normal).
They also had very low concentrations of lutein in the plasma and all other
tissues compared with control chicks. The plasma and retina were only 9% and 6%
of control levels (P < 0.01), respectively. Zeaxanthin levels were similarly low
(9% of control, P < 0.01). The high-lutein diet increased the content of lutein
in the plasma and tissues of control chicks (P < 0.01). In contrast, in WHAM
chicks, lutein increased greatly in the plasma, liver, and heart, but little in
the retina (6% of control). CONCLUSIONS: HDL deficiency in the WHAM chicks was
associated with a deficiency of lutein and zeaxanthin in the tissues, especially
in the retina. The high-lutein diet increased the lutein content of some tissues
via LDL and VLDL transport, but retinal lutein remained very low. These data
support the prime role of HDL as the specific transporter of lutein and
zeaxanthin into the retina. The WHAM chick provides an excellent model for the
study of the role of HDL in the retinal uptake of lutein and zeaxanthin.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17724211 [PubMed - indexed for MEDLINE]

344: Plant Mol Biol. 2007 Oct;65(3):259-75. Epub 2007 Aug 26.

Downstream promoter sequence of an Indian isolate of Rice tungro bacilliform
virus alters tissue-specific expression in host rice and acts differentially in
heterologous system.

Mathur S, Dasgupta I.

Department of Plant Molecular Biology, University of Delhi South Campus, Benito
Juarez Road, New Delhi, 110021, India.

An Indian isolate of Rice tungro bacilliform virus from West Bengal (RTBV-WB)
showed significant nucleotide differences in its putative promoter region when
compared with a previously characterized isolate from Philippines. The
transcription start site of RTBV-WB was mapped followed by assessing the activity
and tissue-specificity of the full-length (FL) promoter (-231 to +645) and
several of its upstream and downstream deletions by studying the expression of
beta-Glucuronidase (GUS) reporter gene in transgenic rice (Oryza sativa L. subsp.
indica) plants at various stages of development. In addition to the expected
vascular-specific expression pattern, studied by histochemical staining, GUS
enzymatic assay and northern and RT-PCR analysis, two novel patterns were
revealed in some of the downstream deleted versions; a non-expressing type,
representing no expression at any stage in any tissue and constitutive type,
representing constitutive expression at all stages in most tissues. This
indicated the presence of previously unreported positive and negative
cis-regulatory elements in the downstream region. The negative element and a
putative enhancer region in the upstream region specifically bound to rice
nuclear proteins in vitro. The FL and its deletion derivatives were also active
in heterologous systems like tobacco (Nicotiana tabacum) and wheat (Triticum
durum). Expression patterns in tobacco were different from those observed in rice
suggesting the importance of upstream elements in those systems and host-specific
regulation of the promoter in diverse organisms. Thus, the RTBV-WB FL promoter
and its derivatives contain an array of cis-elements, which control constitutive
or tissue- and development-specific gene expression in a combinatorial fashion.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17721744 [PubMed - indexed for MEDLINE]

345: Genetics. 2007 Sep;177(1):523-33. Epub 2007 Aug 24.

The expression pattern of a rice disease resistance gene xa3/xa26 is
differentially regulated by the genetic backgrounds and developmental stages that
influence its function.

Cao Y, Ding X, Cai M, Zhao J, Lin Y, Li X, Xu C, Wang S.

National Key Laboratory of Crop Genetic Improvement, National Center of Plant
Gene Research (Wuhan), Huazhong Agricultural University, Wuhan 430070, China.

Genetic background and developmental stage influence the function of some disease
resistance (R) genes. The molecular mechanisms of these modifications remain
elusive. Our results show that the two factors are associated with the expression
of the R gene in rice Xa3 (also known as Xa26)-mediated resistance to Xanthomonas
oryzae pv. oryzae (Xoo), which in turn influences the expression of
defense-responsive genes. The background of japonica rice, one of the two major
subspecies of Asian cultivated rice, facilitates the function of Xa3 more than
the background of indica rice, another rice subspecies. Xa3 expression gradually
increases from early seedling stage to adult stage. Japonica plants carrying Xa3
regulated by the native promoter showed an enlarged resistance spectrum (i.e.,
resistance to more Xoo races), an increased resistance level (i.e., further
reduced lesion length), and whole-growth-stage resistance compared to the indica
rice; this enhanced resistance was associated with an increased expression of Xa3
throughout the growth stages in the japonica plants, which resulted in enhanced
expression of defense-responsive genes. Overexpressing Xa3 with a constitutive
strong promoter further enhanced rice resistance due to further increased Xa3
transcripts in both indica and japonica backgrounds, whereas regulating Xa3 with
a pathogen-induced weak promoter impaired rice resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17720929 [PubMed - indexed for MEDLINE]

346: Regul Toxicol Pharmacol. 2007 Oct;49(1):53-62.

Comparative safety testing of genetically modified foods in a 90-day rat feeding
study design allowing the distinction between primary and secondary effects of
the new genetic event.

Knudsen I, Poulsen M.

Department of Toxicology and Risk Assessment, National Food Institute, Technical
University of Denmark, 19 Moerkhoej Bygade, DK-2860 Soeborg, Denmark.

This article discusses the wider experiences regarding the usefulness of the
90-day rat feeding study for the testing of whole foods from genetically modified
(GM) plant based on data from a recent EU-project [Poulsen, M., Schrö¸der, M.,
Wilcks, A., Kroghsbo, S., Lindecrona, R.H., Miller, A., Frenzel, T., Danier, J.,
Rychlik, M., Shu, Q., Emami, K., Taylor, M., Gatehouse, A., Engel, K.-H.,
Knudsen, I., 2007a. Safety testing of GM-rice expressing PHA-E lectin using a new
animal test design. Food Chem. Toxicol. 45, 364-377; Poulsen, M., Kroghsbo, S.,
Schrö¸der, M., Wilcks, A., Jacobsen, H., Miller, A., Frenzel, T., Danier, J.,
Rychlik, M., Shu, Q., Emami, K., Sudhakar, D., Gatehouse, A., Engel, K.-H.,
Knudsen, I., 2007b. A 90-day safety in Wistar rats fed genetically modified rice
expressing snowdrop lectin Galanthus nivalis (GNA). Food Chem. Toxicol. 45,
350-363; Schrö¸der, M., Poulsen, M., Wilcks, A., Kroghsbo, S., Miller, A.,
Frenzel, T., Danier, J., Rychlik, M., Emami, K., Gatehouse, A., Shu, Q., Engel,
K.-H., Knudsen, I., 2007. A 90-day safety study of genetically modified rice
expressing Cry1Ab protein (Bacillus thuringiensis toxin) in Wistar rats. Food
Chem. Toxicol. 45, 339-349]. The overall objective of the project has been to
develop and validate the scientific methodology necessary for assessing the
safety of foods from genetically modified plants in accordance with the present
EU regulation. The safety assessment in the project is combining the results of
the 90-day rat feeding study on the GM food with and without spiking with the
pure novel gene product, with the knowledge about the identity of the genetic
change, the compositional data of the GM food, the results from in-vitro/ex-vivo
studies as well as the results from the preceding 28-day toxicity study with the
novel gene product, before the hazard characterisation is concluded. The results
demonstrated the ability of the 90-day rat feeding study to detect the
biological/toxicological effects of the new gene product in the GM food. The
authors consider on this basis that the 90-day, rodent feeding study with one
high dose level and a dietary design based upon compositional data on the GM food
and toxicity data on the gene product is sensitive and specific enough to verify
the presence/absence of the biological/nutritional/toxicological effects of the
novel gene insert and further by the use of spiking able to separate potentially
unintended effects of the novel gene product from other unintended effects at the
level of intake defined in the test and within the remit of the test.
Recommendations for further work necessary in the field are given.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17719159 [PubMed - indexed for MEDLINE]

347: Plant Cell Physiol. 2007 Sep;48(9):1263-74. Epub 2007 Aug 23.

Disease resistance against Magnaporthe grisea is enhanced in transgenic rice with
suppression of omega-3 fatty acid desaturases.

Yara A, Yaeno T, Hasegawa M, Seto H, Montillet JL, Kusumi K, Seo S, Iba K.

Department of Biology, Faculty of Sciences, Kyushu University, Hakozaki, Fukuoka,
Japan.

Linolenic acid (18:3) is the most abundant fatty acid in plant membrane lipids
and is a source for various oxidized metabolites, called oxylipins. 18:3 and
oxylipins play important roles in the induction of defense responses to pathogen
infection and wound stress in Arabidopsis. However, in rice, endogenous roles for
18:3 and oxylipins in disease resistance have not been confirmed. We generated
18:3-deficient transgenic rice plants (F78Ri) with co-suppression of two omega-3
fatty acid desaturases, OsFAD7 and OsFAD8. that synthesize 18:3. The F78Ri plants
showed enhanced resistance to the phytopathogenic fungus Magnaporthe grisea. A
typical 18:3-derived oxylipin, jasmonic acid (JA), acts as a signaling molecule
in defense responses to fungal infection in Arabidopsis. However, in F78Ri
plants, the expression of JA-responsive pathogenesis-related genes, PBZ1 and
PR1b, was induced after inoculation with M. grisea, although the JA-mediated
wound response was suppressed. Furthermore, the application of JA methyl ester
had no significant effect on the enhanced resistance in F78Ri plants. Taken
together, our results indicate that, although suppression of fatty acid
desaturases involves the concerted action of varied oxylipins via diverse
metabolic pathways, 18:3 or 18:3-derived oxylipins, except for JA, may contribute
to signaling on defense responses of rice to M. grisea infection.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17716996 [PubMed - indexed for MEDLINE]

348: Int J Food Microbiol. 2007 Oct 20;119(1-2):126-30. Epub 2007 Jul 31.

Strategies for managing Fusarium head blight and deoxynivalenol accumulation in
wheat.

Yuen GY, Schoneweis SD.

Department of Plant Pathology, University of Nebraska-Lincoln, Lincoln, NE
68583-0722, United States. gyuen1@unl.edu

Many mycotoxigenic fungi infect plant hosts and cause disease in the field.
Therefore, control of field infection by these fungi is a critical step in
managing mycotoxin accumulation in the harvested product. Fusarium graminearum,
also known as Gibberella zeae, is the causal agent of Fusarium head blight (FHB),
or scab, in cereals and is also the primary agent responsible for contamination
of grain with deoxynivalenol (DON). Research efforts worldwide are devoted to the
development of strategies to control field infection of wheat and barley by this
pathogen. Strategies include the use of fungicides and biological control agents
to protect flowering heads from infection. There is extensive effort in breeding
for host resistance to infection and spread of the pathogen within the heads.
Scientists are also seeking exogenous traits to introduce into cereals to enhance
resistance. Cultural practices are also being examined, primarily as measures to
reduce pathogen survival and inoculum production in crop residues. The successes
and limitations of these strategies in the management of Fusarium head blight and
deoxynivalenol are discussed.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17716767 [PubMed - indexed for MEDLINE]

349: Int J Food Microbiol. 2007 Oct 20;119(1-2):51-3. Epub 2007 Jul 31.

Marker-assisted selection for Fusarium head blight resistance in wheat.

Anderson JA.

Dept. of Agronomy & Plant Genetics, 411 Borlaug Hall, 1991 Buford Circle,
University of Minnesota, St. Paul, MN 55108, USA. ander319@umn.edu


The cultivation of wheat varieties resistant to Fusarium head blight (FHB) is
recognized as one of the most important components to diminish losses due to this
disease. Although there is no known immunity to this disease in wheat germplasm,
considerable improvements in genetic resistance have been achieved by
concentrated breeding efforts that have relied primarily upon repeated field and
greenhouse-based screening. DNA markers are a relatively new technology that can
be used to increase breeding progress, especially for traits such as FHB that are
difficult to select for under field conditions and that are controlled by
multiple genes. Marker-assisted selection (MAS) uses markers to select for
particular DNA segments that are genetically linked to genes that provide
incremental resistance to FHB. One particular gene, designated Fhb1, provides a
20-25% average reduction in FHB symptoms. This gene and its associated markers
have been validated in numerous breeding programs and is widely used to more
precisely breed for resistance. About a dozen other genes affecting FHB reaction
have been identified, but they have smaller and more inconsistent effects
compared with Fhb1. Nevertheless, breeders are discovering which of these markers
can be combined with Fhb1 in their genetic backgrounds to enhance resistance. The
establishment of the USDA-ARS Regional Small Grains Genotyping Centers and
similar facilities around the world have increased the capacity for wheat
breeders to utilize this powerful technology. More efficient DNA extraction
technologies and marker platforms will allow breeders to more fully implement MAS
in the future.

Publication Types:
Review

PMID: 17716766 [PubMed - indexed for MEDLINE]

350: Environ Entomol. 2007 Aug;36(4):967-81.

Effects of cultivation of genetically modified Bt maize on epigeic arthropods
(Araneae; Carabidae).

Toschki A, Hothorn LA, Ross-Nickoll M.

RWTH-Aachen University, Institute for Environmental Research, Worringer Weg 1,
D-52056 Aachen, Germany. tosch@bio5.rwth-aachen.de

A field study was conducted in Germany to determine the possible effects of
transgenic maize cultivation on nontarget epigeic predator organisms. During the
growing period of 2001-2003, the activity abundances of spiders and carabid
beetles were recorded and compared in three treatments: (1) Bt-maize (Mon 810)
expressing the Cry1ab protein from Bacillus thuringiensis (Berliner), (2) an
isogenic variety, and (3) the isogenic variety treated with insecticide. All
three treatments were replicated in eight plots. The results were evaluated using
three different methods. The activity abundances of single species were
statistically analyzed by confidence interval methods. In addition, the
phenological behaviors of the spider and carabid beetle species were determined,
and multivariate statistical evaluation of the community by principal component
analysis was conducted. Significantly different activity abundances in Bt plots
compared with isogenic control plots were observed both for spiders and carabid
beetles during 2001. However, in 2002 and 2003, no changes in community structure
were detectable in any of the treatments. The change in the first year may have
been caused by the influence of a massive cornborer infestation and accompanying
large changes in microclimatic factors.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17716489 [PubMed - indexed for MEDLINE]

351: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 12;363(1491):557-72.

Marker-assisted selection: an approach for precision plant breeding in the
twenty-first century.

Collard BC, Mackill DJ.

Plant Breeding, Genetics and Biotechnology Division, International Rice Research
Institute (IRRI), DAPO Box 7777, Metro Manila, The Philippines.

DNA markers have enormous potential to improve the efficiency and precision of
conventional plant breeding via marker-assisted selection (MAS). The large number
of quantitative trait loci (QTLs) mapping studies for diverse crops species have
provided an abundance of DNA marker-trait associations. In this review, we
present an overview of the advantages of MAS and its most widely used
applications in plant breeding, providing examples from cereal crops. We also
consider reasons why MAS has had only a small impact on plant breeding so far and
suggest ways in which the potential of MAS can be realized. Finally, we discuss
reasons why the greater adoption of MAS in the future is inevitable, although the
extent of its use will depend on available resources, especially for orphan
crops, and may be delayed in less-developed countries. Achieving a substantial
impact on crop improvement by MAS represents the great challenge for agricultural
scientists in the next few decades.

Publication Types:
Review

PMID: 17715053 [PubMed - indexed for MEDLINE]

352: Plant Biotechnol J. 2007 Nov;5(6):815-26. Epub 2007 Aug 22.

Development of transgenic rice seed accumulating a major Japanese cedar pollen
allergen (Cry j 1) structurally disrupted for oral immunotherapy.

Yang L, Suzuki K, Hirose S, Wakasa Y, Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.

Rice seed-based edible vaccines expressing T-cell epitope peptides derived from
Japanese cedar major pollen allergens have been used to successfully suppress
allergen-specific Th2-mediated immunoglobulin E (IgE) responses in mouse
experiments. In order to further expand the application of seed-based
allergen-specific immunotherapy for controlling Japanese cedar pollinosis, we
generated transgenic rice plants that specifically express recombinant Cry j 1
allergens in seeds. Cry j 1 allergens give low specific IgE-binding activity but
contain all of the T-cell epitopes. The allergens were expressed directly or as a
protein fusion with the major rice storage protein glutelin. Fusion proteins
expressed under the control of the strong rice endosperm-specific GluB-1 promoter
accumulated in rice endosperm tissue up to 15% of total seed protein. The fusion
proteins aggregated with cysteine-rich prolamin and were deposited in endoplasmic
reticulum-derived protein body I. The production of transgenic rice expressing
structurally disrupted Cry j 1 peptides with low IgE binding activity but
spanning the entire Cry j1 region can be used as a universal, safe and effective
tolerogen for rice seed-based oral immunotherapy for cedar pollen allergy in
humans and other mammals.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17714439 [PubMed - indexed for MEDLINE]

353: Plant Mol Biol. 2007 Oct;65(3):329-41. Epub 2007 Aug 22.

A Cre/loxP-mediated self-activating gene excision system to produce marker gene
free transgenic soybean plants.

Li Z, Xing A, Moon BP, Burgoyne SA, Guida AD, Liang H, Lee C, Caster CS, Barton
JE, Klein TM, Falco SC.

DuPont Agriculture & Nutrition, Experimental Station, E353-002C, Route 141 &
Henry Clay Road, Wilmington, DE 19880, USA. Zhongsen.li@cgr.dupont.com

Marker-gene-free transgenic soybean plants were produced by isolating a
developmentally regulated embryo-specific gene promoter, app1, from Arabidopsis
and developing a self-activating gene excision system using the P1 bacteriophage
Cre/loxP recombination system. To accomplish this, the Cre recombinase gene was
placed under control of the app1 promoter and, together with a selectable marker
gene (hygromycin phosphotransferase), were cloned between two loxP recombination
sites. This entire sequence was then placed between a constitutive promoter and a
coding region for either beta-glucuronidase (Gus) or glyphosate acetyltransferase
(Gat). Gene excision would remove the entire sequence between the two loxP sites
and bring the coding region to the constitutive promoter for expression. Using
this system marker gene excision occurred in over 30% of the stable transgenic
events as indicated by the activation of the gus reporter gene or the gat gene in
separate experiments. Transgenic plants with 1 or 2 copies of a functional
excision-activated gat transgene and without any marker gene were obtained in T0
or T1 generation. This demonstrates the feasibility of using developmentally
controlled promoters to mediate marker excision in soybean.

PMID: 17712602 [PubMed - indexed for MEDLINE]

354: Plant Biotechnol J. 2007 Nov;5(6):735-45. Epub 2007 Aug 16.

Enhanced phosphorus nutrition in monocots and dicots over-expressing a
phosphorus-responsive type I H+-pyrophosphatase.

Yang H, Knapp J, Koirala P, Rajagopal D, Peer WA, Silbart LK, Murphy A, Gaxiola
RA.

Department of Plant Science, University of Connecticut, 1390 Storrs Road, Storrs,
CT 06269-4163, USA.

Plants challenged by limited phosphorus undergo dramatic morphological and
architectural changes in their root systems in order to increase their absorptive
surface area. In this paper, it is shown that phosphorus deficiency results in
increased expression of the type I H+-pyrophosphatase AVP1 (AVP, Arabidopsis
vacuolar pyrophosphatase), subsequent increased P-type adenosine triphosphatase
(P-ATPase)-mediated rhizosphere acidification and root proliferation. Molecular
genetic manipulation of AVP1 expression in Arabidopsis, tomato and rice results
in plants that outperform controls when challenged with limited phosphorus.
However, AVP1 over-expression and the resulting rhizosphere acidification do not
result in increased sensitivity to AlPO4, apparently because of the enhancement
of potassium uptake and the release of organic acids. Thus, the over-expression
of type I H+-pyrophosphatases appears to be a generally applicable technology to
help alleviate agricultural losses in low-phosphorus tropical/subtropical soils
and to reduce phosphorus runoff pollution of aquatic and marine environments
resulting from fertilizer application.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17711412 [PubMed - indexed for MEDLINE]

355: Plant Mol Biol. 2007 Nov;65(5):603-14. Epub 2007 Aug 21.

Distinct repressing modules on the distal region of the SBP2 promoter contribute
to its vascular tissue-specific expression in different vegetative organs.

Freitas RL, Carvalho CM, Fietto LG, Loureiro ME, Almeida AM, Fontes EP.

Departamento de Biologia Vegetal, Universidade Federal de Viö§osa, 36571-000
Vicosa, MG, Brazil.

The Glycine max sucrose binding protein (GmSBP2) promoter directs vascular
tissue-specific expression of reporter genes in transgenic tobacco. Here we
showed that an SBP2-GFP fusion protein under the control of the GmSBP2 promoter
accumulates in the vascular tissues of vegetative organs, which is consistent
with the proposed involvement of SBP in sucrose transport-dependent physiological
processes. Through gain-of-function experiments we confirmed that the
tissue-specific determinants of the SBP2 promoter reside in the distal
cis-regulatory domain A, CRD-A (position -2000 to -700) that is organized into a
modular configuration to suppress promoter activity in tissues other than
vascular tissues. The four analyzed CRD-A sub-modules, designates Frag II
(-1785/-1508), Frag III (-1507/-1237), Frag IV (-1236/-971) and Frag V
(-970/-700), act independently to alter the constitutive pattern of
-92pSBP2-mediated GUS expression in different organs. Frag V fused to
-92pSBP2-GUS restored the tissue-specific pattern of the full-length promoter in
the shoot apex, but not in other organs. Likewise, Frag IV confined GUS
expression to the vascular bundle of leaves, whereas Frag II mediated vascular
specific expression in roots. Strong stem expression-repressing elements were
located at positions -1485 to -1212, as Frag III limited GUS expression to the
inner phloem. We have also mapped a procambium silencer to the consensus sequence
CAGTTnCaAccACATTcCT which is located in both distal and proximal upstream
modules. Fusion of either repressing element-containing module to the
constitutive -92pSBP2 promoter suppresses GUS expression in the elongation zone
of roots. Together our results demonstrate the unusual aspect of distal sequences
negatively controlling tissue-specificity of a plant promoter.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17710554 [PubMed - indexed for MEDLINE]

356: Mol Cell Neurosci. 2007 Oct;36(2):211-21. Epub 2007 Jul 24.

The Drosophila ARC homolog regulates behavioral responses to starvation.

Mattaliano MD, Montana ES, Parisky KM, Littleton JT, Griffith LC.

Department of Biology and National Center for Behavior Genomics, Brandeis
University, Waltham, MA 02454-9110, USA.

The gene encoding dARC1, one of three Drosophila homologs of mammalian
activity-regulated cytoskeleton-associated protein (ARC), is upregulated in both
seizure and muscular hypercontraction mutants. In this study we generate a null
mutant for dArc1 and show that this gene is not involved in synaptic plasticity
at the larval neuromuscular junction or in formation or decay of short-term
memory of courtship conditioning, but rather is a modifier of stress-induced
behavior. dARC1 is expressed in a number of neurosecretory cells and mutants are
starvation-resistant, exhibiting an increased time of survival in the absence of
food. Starvation resistance is likely due to the fact that dArc1 mutants lack the
normal hyperlocomotor response to starvation, which is almost universal in the
animal kingdom. dARC1 acts in insulin-producing neurons of the pars
intercerebralis to control this behavior, but does not appear to be a general
regulator of insulin signaling. This suggests that there are multiple modes of
communication between the pars and the ring gland that control starvation-induced
behavioral responses.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17707655 [PubMed - indexed for MEDLINE]

357: Phytochemistry. 2008 Jan;69(1):76-87. Epub 2007 Aug 17.

Inactivation of pea genes by RNAi supports the involvement of two similar
O-methyltransferases in the biosynthesis of (+)-pisatin and of chiral
intermediates with a configuration opposite that found in (+)-pisatin.

Kaimoyo E, VanEtten HD.

Division of Plant Pathology and Microbiology, Department of Plant Sciences, 1140
E. South Campus Drive, Forbes 303, University of Arizona, Tucson, AZ 85721,
United States.

(+)-Pisatin, the major phytoalexin of pea (Pisum sativum L.), is believed to be
synthesized via two chiral intermediates,
(-)-7,2'-dihydroxy-4',5'-methylenedioxyisoflavanone [(-)-sophorol] and
(-)-7,2'-dihydroxy-4',5'-methylenedioxyisoflavanol [(-)-DMDI]; both have an
opposite C-3 absolute configuration to that found at C-6a in (+)-pisatin. The
expression of isoflavone reductase (IFR), which converts
7,2'-dihydroxy-4',5'-methylenedioxyisoflavone (DMD) to (-)-sophorol, sophorol
reductase (SOR), which converts (-)-sophorol to (-)-DMDI, and
hydroxymaackiain-3-O-methyltransferase (HMM), believed to be the last step of
(+)-pisatin biosynthesis, were inactivated by RNA-mediated genetic interference
(RNAi) in pea hairy roots. Some hairy root lines containing RNAi constructs of
IFR and SOR accumulated DMD or (-)-sophorol, respectively, and were deficient in
(+)-pisatin biosynthesis supporting the involvement of chiral intermediates with
a configuration opposite to that found in (+)-pisatin in the biosynthesis of
(+)-pisatin. Pea proteins also converted (-)-DMDI to an achiral isoflavene
suggesting that an isoflavene might be the intermediate through which the
configuration is changed to that found in (+)-pisatin. Hairy roots containing
RNAi constructs of HMM also were deficient in (+)-pisatin biosynthesis, but did
not accumulate (+)-6a-hydroxymaackiain, the proposed precursor to (+)-pisatin.
Instead, 2,7,4'-trihydroxyisoflavanone (TIF), daidzein, isoformononetin, and
liquiritigenin accumulated. HMM has a high amino acid similarity to
hydroxyisoflavanone-4'-O-methyltransferase (HI4'OMT), an enzyme that methylates
TIF, an early intermediate in the isoflavonoid pathway. The accumulation of these
four compounds is consistent with the blockage of the synthesis of (+)-pisatin at
the HI4'OMT catalyzed step resulting in the accumulation of liquiritigenin and
TIF and the diversion of the pathway to produce daidzein and isoformononetin,
compounds not normally made by pea. Previous results have identified two highly
similar "HMMs" in pea. The current results suggest that both of these
O-methyltransferases are involved in (+)-pisatin biosynthesis and that one
functions early in the pathway as HI4'OMT and the second acts at the terminal
step of the pathway.

PMID: 17707445 [PubMed - indexed for MEDLINE]

358: Poult Sci. 2007 Sep;86(9):1988-94.

Comparison of broiler performance and carcass parameters when fed diets
containing combined trait insect-protected and glyphosate-tolerant corn (MON
89034 x NK603), control, or conventional reference corn.

Taylor M, Lucas D, Nemeth M, Davis S, Hartnell G.

Monsanto Company, Product Safety Center, Creve Couer, MO 63167, USA.
mary.l.taylor@monsanto.com

A 42-d floor pen study was conducted to compare broiler (Ross x Ross 308)
performance and carcass measurements when fed diets containing
lepidopteran-protected corn combined with glyphosate-tolerant corn (MON 89034 x
NK603) with those of broilers fed diets containing corn grain from the
conventional control (similar genetic background to the test corn) and 6
conventional corn hybrids. It has been found that MON 89034 produces the
Cry1A.105 and Cry2Ab2 insecticidal proteins that protect corn plants from feeding
damage caused by European corn borer (Ostrinia nubilalis) and other lepidopteran
insect pests. In addition, NK603 produces the 5-enolpyruvylshikimate-3-phosphate
synthase protein from Agrobacterium sp. strain CP4 (CP4 EPSPS), which confers
tolerance to glyphosate, the active ingredient in Roundup agricultural
herbicides. The traditional breeding of plants that express the individual traits
produced MON 89034 x NK603. Broilers were fed starter diets (approximately 57%
wt/wt corn grain) from d 0 to 21 and grower-finisher diets (approximately 59%
wt/wt corn grain) from d 21 to 42. The study utilized a randomized complete block
design with 8 dietary treatments assigned randomly within 5 blocks of 16 pens
each (8 male and 8 female) with 10 birds per pen. There were 10 pens per
treatment group (5 male and 5 female). Weight at d 0 and 42, feed intake, feed
conversion, and all measured carcass and meat quality parameters were not
different (P > 0.05) for birds fed MON 89034 x NK603 and control corn diets. In
addition, comparisons of the MON 89034 x NK603 diet to the population of the
control and 6 reference corn diets showed no difference (P > 0.05) in any
performance, carcass, or meat quality parameter measured. In conclusion, the
diets containing MON 89034 x NK603 were nutritionally equivalent to diets
containing the control or conventional reference corn grain when fed to broilers.

Publication Types:
Comparative Study
Randomized Controlled Trial

PMID: 17704388 [PubMed - indexed for MEDLINE]

359: Poult Sci. 2007 Sep;86(9):1972-9.

Comparison of broiler performance when fed diets containing grain from
second-generation insect-protected and glyphosate-tolerant, conventional control
or commercial reference corn.

Taylor M, Hartnell G, Nemeth M, Lucas D, Davis S.

Monsanto Co., Product Safety Center, Creve Coeur, MO 63167, USA.
mary.l.taylor@monsanto.com

Two 42-d floor pen studies were conducted to compare broiler (Ross x Ross 308)
performance and carcass measurements when broilers were fed diets containing
grain from either second-generation lepidopteran insect-protected corn (MON
89034; study 1) or second-generation lepidopteran combined with second-generation
corn rootworm-protected and glyphosate-tolerant corn (MON 89034 x MON 88017;
study 2) with those of diets containing corn grain from the conventional control
and 4 conventional corn hybrids. In both studies, broilers were fed starter diets
(approximately 55%, wt/wt, corn grain) from d 0 to 21 and grower-finisher diets
(approximately 59%, wt/wt, corn grain) from d 21 to 42. Each study used a
randomized complete block design with 6 dietary treatments assigned randomly
within 5 blocks of 12 pens each (6 male and 6 female) and 10 pens per treatment
group (5 male and 5 female). In study 1 (MON 89034), no treatment differences
were detected among dietary treatments for feed intake, weight gain, or any
measured carcass parameter. A significant difference was noted for adjusted feed
conversion between MON 89034 and control birds; however, no differences were
detected in individual treatment comparisons between the MON 89034 and 3 of the 4
commercial corn diets. In study 2 (MON 89034 x MON 88017), no treatment
differences were observed for feed intake and most carcass parameters. When
significant treatment differences were detected, no differences were observed
between MON 89034 x MON 88017, its control, and 2 or more of the commercial corn
diets. In each study, comparison of all parameters measured showed no differences
between birds fed the test diet and the population of birds fed the control and 4
commercial corn diets. In conclusion, the test diets were nutritionally
equivalent to diets containing the control and corn grain from commercial
hybrids.

Publication Types:
Comparative Study
Randomized Controlled Trial

PMID: 17704386 [PubMed - indexed for MEDLINE]

360: Sci Eng Ethics. 2007 Mar;13(1):69-82.

A case for a duty to feed the hungry: GM plants and the third world.

Carter L.

The School of History, Philosophy, Religion and Classics, The University of
Queensland, Brisbane, Australia. l.carter@uq.edu.au

This article is concerned with a discussion of the plausibility of the claim that
GM technology has the potential to provide the hungry with sufficient food for
subsistence. Following a brief outline of the potential applications of GM in
this context, a history of the green revolution and its impact will be discussed
in relation to the current developing world agriculture situation. Following a
contemporary analysis of malnutrition, the claim that GM technology has the
potential to provide the hungry with sufficient nourishment will be discussed
within the domain of moral philosophy to determine whether there exists a moral
obligation to pursue this end if and only if the technology proves to be
relatively safe and effective. By using Peter Singer's duty of moral rescue, I
argue that we have a moral duty to assist the third world through the
distribution of such GM plants. I conclude the paper by demonstrating that my
argument can be supported by applying a version of the Precautionary Principle on
the grounds that doing nothing might be worse for the current situation.

PMID: 17703610 [PubMed - indexed for MEDLINE]

361: Biotechnol J. 2007 Sep;2(9):1086-7.

The difficulty of structuring and focusing the co-existence debate in Europe.

Custers R.

VIB, Gent, Belgium. Rene.Custers@vib.be

The co-existence debate in Europe is wide and difficult. In this paper some
recommendations are given on how to make progress in the debate. Not with the
goal of pushing GMOs, but with the goal of achieving genuine freedom of choice.

PMID: 17703493 [PubMed - indexed for MEDLINE]

362: Biotechnol J. 2007 Sep;2(9):1141-6.

Transparent communication strategy on GMOs: will it change public opinion?

Sinemus K, Egelhofer M.

Genius Gmbh, Darmstadt, Germany. Kristina.Sinemus@genius.de

Innovations are central for the economic growth; however, the use of new
technologies needs to be widely accepted in the general public and the society as
a whole. Biotechnology in general, and the use of genetic engineering in food
production in particular are seen critically by the European public and perceived
as "risky", and a transatlantic divide between European and US citizens has been
observed. This review investigates the reasons for those differing perceptions
and proposes new strategies to communicate the benefits of biotechnology in
agriculture to a broader public. When analyzing the dialogue process that has
taken place between public, scientists, governmental organizations and industry,
questions arise on what has been done differently in Europe, in order to propose
new, more successful and efficient communication strategies for the future.

Publication Types:
Review

PMID: 17703492 [PubMed - indexed for MEDLINE]

363: Biotechnol J. 2007 Sep;2(9):1088-92.

Intellectual property, genetically modified crops and bioethics.

Adcock M.

Department of Law, Durham University, Durham, UK. mike.adcock@durham.ac.uk

The implementation of a new technology is almost always surrounded by a debate on
the moral and social implications that may arise. The debate with regard to
genetically modified (GM) crops has been one of the longest and most
controversial. However, one area of the debate that receives less attention is
the role that intellectual property can play. The introduction of an effective
and yet appropriate intellectual property system addressing society's particular
needs can eliminate some of these issues. This paper looks at whether the
situation in Europe is meeting our current needs and also addresses the role
intellectual property can play in the debate over the introduction of GM crops in
developing countries.

Publication Types:
Review

PMID: 17703487 [PubMed - indexed for MEDLINE]

364: Br Poult Sci. 2007 Aug;48(4):480-8.

The effects of xylanase supplementation on growth, digestion, circulating hormone
and metabolite levels, immunity and gut microflora in cockerels fed on
wheat-based diets.

Gao F, Jiang Y, Zhou GH, Han ZK.

College of Animal Science and Technology, Nanjing Agricultural University,
Nanjing, Jiangsu, China. gaofeng0629@sina.com

1. The xylanase product used in this study was derived from a genetically
modified isolate of Aspergillus niger. Two trials were conducted to investigate
the effects of xylanase supplementation on growth, digestion, circulating hormone
and metabolite levels, immune parameters and composition of the gut microflora in
cockerels fed on wheat-based diets. 2. The experimental diets consisted of a
wheat-based control diet supplemented with 0 or 0.1% enzyme preparation. The
diets were fed between 7 and 21 d of age. 3. Enzyme supplementation improved
growth and feed conversion efficiency. The addition of enzyme to wheat-based diet
increased the apparent total digestibility of dry matter (DM), crude protein and
fat. 4. Enzyme supplementation reduced the relative weight of digestive organs to
a certain extent, but there was no significant difference. Enzyme supplementation
reduced digesta viscosity in the jejunum. There was no significant difference
between the two experimental groups in counts of lactobacillus and coliform
bacteria in the caeca. 5. Enzyme supplementation increased the concentration of
blood thyroxine (T(4)), insulin-like growth factor I (IGF-I) and insulin, reduced
the concentrations of blood uric acid, but had no significant effect on the
concentrations of blood glucose and triiodothyronine (T(3)). 6. Enzyme
supplementation increased the relative weight of spleen of cockerels, serum
antibody titres to Newcastle disease virus (NDV), lymphocyte proliferation in
response to phytohaemagglutinin (PHA) and the natural killer (NK) cell activity.
7. It is concluded that supplementation with an enzyme preparation (xylanase),
which hydrolyses non-starch polysaccharides can improve growth in cockerels fed
on wheat-based diets. This improvement is achieved through enzyme effects on
digestion, absorption, metabolism and immunity of cockerels.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17701501 [PubMed - indexed for MEDLINE]

365: Transgenic Res. 2007 Dec;16(6):675-88. Epub 2007 Aug 14.

Genetically modified crops for the bioeconomy: meeting public and regulatory
expectations.

Chapotin SM, Wolt JD.

US Agency for International Development, Washington, DC 20523, USA.

As the United States moves toward a plant-based bioeconomy, a large research and
development effort is focused on creating new feedstocks to meet biomass demand
for biofuels, bioenergy, and specialized bioproducts, such as industrial
compounds and biomaterial precursors. Most bioeconomy projections assume the
widespread deployment of novel feedstocks developed through the use of modern
molecular breeding techniques, but rarely consider the challenges involved with
the use of genetically modified crops, which can include hurdles due to
regulatory approvals, market adoption, and public acceptance. In this paper we
consider the implications of various transgenic crops and traits under
development for the bioeconomy that highlight these challenges. We believe that
an awareness of the issues in crop and trait selection will allow developers to
design crops with maximum stakeholder appeal and with the greatest potential for
widespread adoption, while avoiding applications unlikely to meet regulatory
approval or gain market and public acceptance.

Publication Types:
Review

PMID: 17701080 [PubMed - indexed for MEDLINE]

366: Nature. 2007 Aug 16;448(7155):736.

Geneticist trades plants for politics. Nina Fedoroff interviewed by Emma Marris.

Fedoroff N.

Publication Types:
Interview

PMID: 17700665 [PubMed - indexed for MEDLINE]

367: J Dairy Sci. 2007 Sep;90(9):4084-91.

Use of human lysozyme transgenic goat milk in cheese making: effects on lactic
acid bacteria performance.

Scharfen EC, Mills DA, Maga EA.

Department of Animal Science, University of California, Davis 95616, USA.

Genetically engineered goats expressing elevated levels of the antimicrobial
enzyme lysozyme in their milk were developed to improve udder health, product
shelf life, and consumer well-being. The purpose of this study was to evaluate
the effect of lysozyme on the development of lactic acid bacteria (LAB)
throughout the cheese-making process. Raw and pasteurized milk from 7 lysozyme
transgenic goats and 7 breed-, age-, and parity-matched nontransgenic controls
was transformed into cheeses by using industry methods, and their microbiological
load was evaluated. The numbers of colony-forming units of LAB were determined
for raw and pasteurized goat milk, whey, and curd at d 2 and at d 6 or 7 of
production. Selective plating media were used to enumerate lactococcal species
separately from total LAB. Although differences in the mean number of
colony-forming units between transgenic and control samples in raw milk, whey,
and cheese curd were non-significant for both total LAB and lactococcal species
from d 2 of production, a significant decrease was observed in both types of LAB
among d 6 transgenic raw milk cheese samples. In pasteurized milk trials, a
significant decrease in LAB was observed only in the raw milk of transgenic
animals. These results indicate that lysozyme transgenic goat milk is not
detrimental to LAB growth during the cheese-making process.

PMID: 17699025 [PubMed - indexed for MEDLINE]

368: J Dairy Sci. 2007 Sep;90(9):4005-21.

Invited review: Advances in starter cultures and cultured foods.

Cogan TM, Beresford TP, Steele J, Broadbent J, Shah NP, Ustunol Z.

Moorepark Food Reseach Centre, Teagasc, Fermoy, Ireland.

With 2005 retail sales close to $4.8 million, cultured dairy products are driving
the growth of dairy foods consumption. Starter cultures are of great industrial
significance in that they play a vital role in the manufacturing, flavor, and
texture development of fermented dairy foods. Furthermore, additional interest in
starter bacteria has been generated because of the data accumulating on the
potential health benefits of these organisms. Today, starter cultures for
fermented foods are developed mainly by design rather than by the traditional
screening methods and trial and error. Advances in genetics and molecular biology
have provided opportunities for genomic studies of these economically significant
organisms and engineering of cultures that focuses on rational improvement of the
industrially useful strain. Furthermore, much research has been published on the
health benefits associated with ingesting cultured dairy foods and probiotics,
particularly their role in modulating immune function. The aim of this review is
to describe some of the major scientific advances made in starter and non-starter
lactic acid bacteria during the past 10 yr, including genomic studies on dairy
starter cultures, engineering of culture attributes, advances in phage control,
developments in methods to enumerate lactic acid bacteria and probiotics in dairy
foods, and the potential role of cultured dairy foods in modulation of immune
function.

Publication Types:
Review

PMID: 17699017 [PubMed - indexed for MEDLINE]

369: Neuron. 2007 Aug 16;55(4):662-76.

Dopamine mediates context-dependent modulation of sensory plasticity in C.
elegans.

Kindt KS, Quast KB, Giles AC, De S, Hendrey D, Nicastro I, Rankin CH, Schafer WR.

Biomedical Sciences Graduate Program, University of California, San Diego, La
Jolla CA 92093, USA.

Dopamine has been implicated in the modulation of diverse forms of behavioral
plasticity, including appetitive learning and addiction. An important challenge
is to understand how dopamine's effects at the cellular level alter the
properties of neural circuits to modify behavior. In the nematode C. elegans,
dopamine modulates habituation of an escape reflex triggered by body touch. In
the absence of food, animals habituate more rapidly than in the presence of food;
this contextual information about food availability is provided by dopaminergic
mechanosensory neurons that sense the presence of bacteria. We find that dopamine
alters habituation kinetics by selectively modulating the touch responses of the
anterior-body mechanoreceptors; this modulation involves a D1-like dopamine
receptor, a Gq/PLC-beta signaling pathway, and calcium release within the touch
neurons. Interestingly, the body touch mechanoreceptors can themselves excite the
dopamine neurons, forming a positive feedback loop capable of integrating context
and experience to modulate mechanosensory attention.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17698017 [PubMed - indexed for MEDLINE]

370: BMC Biotechnol. 2007 Aug 14;7:47.

A minimal peach type II chlorophyll a/b-binding protein promoter retains
tissue-specificity and light regulation in tomato.

Bassett CL, Callahan AM, Artlip TS, Scorza R, Srinivasan C.

US Department of Agriculture, the Agricultural Research Service, Appalachian
Fruit Research Station, 2217 Wiltshire Road, Kearneysville, West Virginia 25430,
USA. Carole.Bassett@ars.usda.gov

BACKGROUND: Promoters with tissue-specificity are desirable to drive expression
of transgenes in crops to avoid accumulation of foreign proteins in edible
tissues/organs. Several photosynthetic promoters have been shown to be strong
regulators of expression of transgenes in light-responsive tissues and would be
good candidates for leaf and immature fruit tissue-specificity, if expression in
the mature fruit were minimized. RESULTS: A minimal peach chlorophyll a/b-binding
protein gene (Lhcb2*Pp1) promoter (Cab19) was isolated and fused to an uidA
(beta-glucuronidase [GUS]) gene containing the PIV2 intron. A control vector
carrying an enhanced mas35S CaMV promoter fused to uidA was also constructed. Two
different orientations of the Cab19::GUS fusion relative to the left T-DNA border
of the binary vector were transformed into tomato. Ten independent regenerants of
each construct and an untransformed control line were assessed both qualitatively
and quantitatively for GUS expression in leaves, fruit and flowers, and
quantitatively in roots. CONCLUSION: The minimal CAB19 promoter conferred GUS
activity primarily in leaves and green fruit, as well as in response to light.
GUS activity in the leaves of both Cab19 constructs averaged about 2/3 that
observed with mas35S::GUS controls. Surprisingly, GUS activity in transgenic
green fruit was considerably higher than leaves for all promoter constructs;
however, in red, ripe fruit activities were much lower for the Cab19 promoter
constructs than the mas35S::GUS. Although GUS activity was readily detectable in
flowers and roots of mas35S::GUStransgenic plants, little activity was observed
in plants carrying the Cab19 promoter constructs. In addition, the
light-inducibility of the Cab19::GUS constructs indicated that all the requisite
cis-elements for light responsiveness were contained on the Cab19 fragment. The
minimal Cab19 promoter retains both tissue-specificity and light regulation and
can be used to drive expression of foreign genes with minimal activity in mature,
edible fruit.

PMID: 17697347 [PubMed - indexed for MEDLINE]

371: Planta. 2007 Dec;227(1):199-209. Epub 2007 Aug 11.

Specific role of LeMAN2 in the control of seed germination exposed by
overexpression of the LeMAN3 gene in tomato plants.

Belotserkovsky H, Berger Y, Shahar R, Wolf S.

Faculty of Agricultural, Food and Environmental Quality Sciences, The Institute
of Plant Sciences and Genetics in Agriculture and the Otto Warburg Minerva Center
for Agricultural Biotechnology, The Hebrew University of Jerusalem, Rehovot,
Israel.

Endo-beta-mannanase is one of the key enzymes involved in the hydrolysis of the
mannan-rich cell walls of tomato (Solanum lycopersicon) seeds. Two isoforms of
endo-beta-mannanase have been characterized in tomato seeds: LeMAN2 is active in
the micropylar area prior to germination and LeMAN1 is active after germination
in all endosperm cells surrounding the cotyledons. To explore whether general
mannanase activity in the endosperm cap is sufficient to promote germination, the
gene encoding LeMAN3 was inserted into transgenic tomato plants under the control
of a CaMV-35S promoter. Expression of LeMAN3 was evident in the endosperm cap and
in the lateral endosperm of the transgenic seeds 10 min after imbibition. An
activity test indicated increased activity of endo-beta-mannanase in the
transgenic lines relative to the control line in all seed parts, during the first
20 h of imbibition. However, overexpression of LeMAN3 in transgenic seeds
inhibited seed germination at both optimal and suboptimal temperatures. Detailed
RT-PCR analyses revealed the transcription patterns of the genes encoding the
various mannanase isoforms, and indicated a delay in LeMAN2 transcription in the
endosperm cap of the transgenic seeds. Interestingly, tissue-print assays
indicated similar mannanase activity in the micropylar areas for both transgenic
and control seeds. These results indicate that overexpression of active
endo-beta-mannanase in the endosperm cap is not sufficient to enable hydrolysis
of the cell walls or to promote germination of tomato seeds. Cell-wall hydrolysis
in these endosperm cells is under tight control and requires the specific
activity of LeMAN2.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17694319 [PubMed - indexed for MEDLINE]

372: Plant Mol Biol. 2007 Oct;65(3):285-94. Epub 2007 Aug 11.

Spatial and temporal regulation of the forisome gene for1 in the phloem during
plant development.

Noll GA, Fontanellaz ME, Rö¼ping B, Ashoub A, van Bel AJ, Fischer R, Knoblauch M,
Prö¼fer D.

Institut fö¼r Biochemie und Biotechnologie der Pflanzen der, Westfö¤lischen
Wilhelms-Universitö¤t Mö¼nster, Hindenburgplatz 55, 48143, Munster, Germany.

Forisomes are protein aggregates found uniquely in the sieve elements of
Fabaceaen plants. Upon wounding they undergo a reversible, calcium-dependent
conformational switch which enables them to act as cellular stopcocks. Forisomes
begin to form in young sieve elements at an early stage of metaphloem
differentiation. Genes encoding forisome components could therefore be useful as
markers of early sieve element development. Here we present a comprehensive
analysis of the developmental expression profile of for1, which encodes such a
forisome component. The for1 gene is highly conserved among Fabaceaen species and
appears to be unique to this phylogenetic lineage since no orthologous genes have
been found in other plants, including Arabidopsis and rice. Even so, transgenic
tobacco plants expressing reporter genes under the control of the for1 promoter
display reporter activity exclusively in immature sieve elements. This suggests
that the regulation of sieve element development is highly conserved even in
plants where mature forisomes have not been detected. The promoter system could
therefore provide a powerful tool for the detailed analysis of differentiation in
metaphloem sieve elements in an unexpectedly broad range of plant species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17694275 [PubMed - indexed for MEDLINE]

373: Trends Plant Sci. 2007 Sep;12(9):397-403. Epub 2007 Aug 10.

The intragenic approach as a new extension to traditional plant breeding.

Rommens CM, Haring MA, Swords K, Davies HV, Belknap WR.

Simplot Plant Sciences, J. R. Simplot Company, Boise, ID 83706, USA.
crommens@simplot.com

The novel intragenic approach to genetic engineering improves existing varieties
by eliminating undesirable features and activating dormant traits. It transforms
plants with native expression cassettes to fine-tune the activity and/or tissue
specificity of target genes. Any intragenic modification of traits could, at
least in theory, also be accomplished by traditional breeding and transgenic
modification. However, the new approach is unique in avoiding the transfer of
unknown or foreign DNA. By consequently eliminating various potential risk
factors, this method represents a relatively safe approach to crop improvement.
Therefore, we argue that intragenic crops should be cleared through the
regulatory process in a timely and cost-effective manner.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17692557 [PubMed - indexed for MEDLINE]

374: Genomics. 2007 Oct;90(4):516-29. Epub 2007 Aug 9.

The alpha- and beta-expansin and xyloglucan endotransglucosylase/hydrolase gene
families of wheat: molecular cloning, gene expression, and EST data mining.

Liu Y, Liu D, Zhang H, Gao H, Guo X, Wang D, Zhang X, Zhang A.

State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of
Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101,
People's Republic of China.

Expansins and xyloglucan endotransglucosylase/hydrolases (XTHs) are families of
extracellular proteins with members that have been shown to play an important
role in cell wall growth. In this study, three, six, and five members of the
wheat alpha-expansin (TaEXPA1 to TaEXPA3), beta-expansin (TaEXPB1 to TaEXPB6),
and XTH (TaXTH1 to TaXTH5) gene families, respectively, were isolated from a
dwarf wheat line. The mRNA expression analysis by real-time RT-PCR indicates that
these genes display different transcription levels in different
stages/organs/treatments, possibly suggesting their functional roles in the cell
wall expansion process. Moreover, the comparison of the expression levels reveals
that most of the expansins show lower expression than the XTHs. Finally, we
present the analysis of wheat alpha- and beta-expansins and XTH families by
expressed sequence tag data mining.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17692501 [PubMed - indexed for MEDLINE]

375: Food Chem Toxicol. 2007 Dec;45(12):2513-25. Epub 2007 Jun 21.

History of safe use as applied to the safety assessment of novel foods and foods
derived from genetically modified organisms.

Constable A, Jonas D, Cockburn A, Davi A, Edwards G, Hepburn P, Herouet-Guicheney
C, Knowles M, Moseley B, Oberdö¶rfer R, Samuels F.

Nestlö© Research Centre, Vers-Chez-les-blanc 1000, Lausanne 26, Switzerland.

Very few traditional foods that are consumed have been subjected to systematic
toxicological and nutritional assessment, yet because of their long history and
customary preparation and use and absence of evidence of harm, they are generally
regarded as safe to eat. This 'history of safe use' of traditional foods forms
the benchmark for the comparative safety assessment of novel foods, and of foods
derived from genetically modified organisms. However, the concept is hard to
define, since it relates to an existing body of information which describes the
safety profile of a food, rather than a precise checklist of criteria. The term
should be regarded as a working concept used to assist the safety assessment of a
food product. Important factors in establishing a history of safe use include:
the period over which the traditional food has been consumed; the way in which it
has been prepared and used and at what intake levels; its composition and the
results of animal studies and observations from human exposure. This paper is
aimed to assist food safety professionals in the safety evaluation and regulation
of novel foods and foods derived from genetically modified organisms, by
describing the practical application and use of the concept of 'history of safe
use'.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17692450 [PubMed - indexed for MEDLINE]

376: Plant Biotechnol J. 2007 Nov;5(6):768-77. Epub 2007 Aug 13.

Genetically stable expression of functional miraculin, a new type of alternative
sweetener, in transgenic tomato plants.

Sun HJ, Kataoka H, Yano M, Ezura H.

Graduate School of Life and Environmental Sciences, Gene Research Center,
University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-8572, Japan.

Miraculin is a taste-modifying protein isolated from the red berries of
Richadella dulcifica, a shrub native to West Africa. Miraculin by itself is not
sweet, but it is able to turn a sour taste into a sweet taste. This unique
property has led to increasing interest in this protein. In this article, we
report the high-yield production of miraculin in transgenic tomato plants. High
and genetically stable expression of miraculin was confirmed by Western blot
analysis and enzyme-linked immunosorbent assay. Recombinant miraculin accumulated
to high levels in leaves and fruits, up to 102.5 and 90.7 microg/g fresh weight,
respectively. Purified recombinant miraculin expressed in transgenic tomato
plants showed strong sweetness-inducing activity, similar to that of native
miraculin. These results demonstrate that recombinant miraculin was correctly
processed in transgenic tomato plants, and that this production system could be a
good alternative to production from the native plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17692073 [PubMed - indexed for MEDLINE]

377: Plant Mol Biol. 2007 Nov;65(4):453-66. Epub 2007 Aug 10.

T-DNA tagged knockout mutation of rice OsGSK1, an orthologue of Arabidopsis BIN2,
with enhanced tolerance to various abiotic stresses.

Koh S, Lee SC, Kim MK, Koh JH, Lee S, An G, Choe S, Kim SR.

Department of Life Science, Sogang University, Seoul 121-742, Korea.

T-DNA-tagged rice plants were screened under cold- or salt-stress conditions to
determine the genes involved in the molecular mechanism for their abiotic-stress
response. Line 0-165-65 was identified as a salt-responsive line. The gene
responsible for this GUS-positive phenotype was revealed by inverse PCR as OsGSK1
(Oryza sativa glycogen synthase kinase3-like gene 1), a member of the plant
GSK3/SHAGGY-like protein kinase genes and an orthologue of the Arabidopsis
brassinosteroid insensitive 2 (BIN2), AtSK21. Northern blot analysis showed that
OsGSK1 was most highly detected in the developing panicles, suggesting that its
expression is developmental stage specific. Knockout (KO) mutants of OsGSK1
showed enhanced tolerance to cold, heat, salt, and drought stresses when compared
with non-transgenic segregants (NT). Overexpression of the full-length OsGSK1 led
to a stunted growth phenotype similar to the one observed with the
gain-of-function BIN/AtSK21 mutant. This suggests that OsGSK1 might be a
functional rice orthologue that serves as a negative regulator of brassinosteroid
(BR)-signaling. Therefore, we propose that stress-responsive OsGSK1 may have
physiological roles in stress signal-transduction pathways and floral
developmental processes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17690841 [PubMed - indexed for MEDLINE]

378: Virus Res. 2007 Dec;130(1-2):210-27. Epub 2007 Aug 8.

Effectiveness and stability of heterologous proteins expressed in plants by
Turnip mosaic virus vector at five different insertion sites.

Chen CC, Chen TC, Raja JA, Chang CA, Chen LW, Lin SS, Yeh SD.

Department of Plant Pathology, National Chung-Hsing University, Taichung 40227,
Taiwan, ROC.

The N-terminal (NT) regions of particular protein-coding sequences are generally
used for in-frame insertion of heterologous open reading frames (ORFs) in
potyviral vectors for protein expression in plants. An infectious cDNA clone of
Turnip mosaic virus (TuMV) isolate YC5 was engineered at the generally used NT
regions of HC-Pro and CP, and other possibly permissive sites to investigate
their effectiveness to express the GFP (jellyfish green fluorescent protein) and
Der p 5 (allergen from the dust mite, Dermatophagoides pteronyssinus) ORFs. The
results demonstrated the permissiveness of the NT regions of P3, CIP and NIb to
carry the ORFs and express the translates as part of the viral polyprotein, the
processing of which released free-form proteins in the host cell milieu. However,
these sites varied in their permissiveness to retain the ORFs intact and hence
affect the heterologous protein expression. Moreover, strong influence of the
inserted ORF and host plants in determining the permissiveness of a viral genomic
context to stably carry the alien ORFs and hence to support their prolonged
expression was also noticed. In general, the engineered sites were relatively
more permissive to the GFP ORF than to the Der p 5 ORF. Among the hosts, the
local lesion host, Chenopodium quinoa Willd. showed the highest extent of support
to TuMV to stably carry the heterologous ORFs at the engineered sites and the
protein expression therefrom. Among the systemic hosts, Nicotiana benthamiana
Domin proved more supportive to TuMV to carry and express the heterologous ORFs
than the Brassica hosts, whereas the protein expression levels were significantly
higher and more stable in the plants of Brassica campestris L. var. chinensis and
B. campestris L. var. ching-geeng than those in the plants of B. juncea L. and B.
campestris L. var. pekinensis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17689817 [PubMed - indexed for MEDLINE]

379: New Phytol. 2007;175(4):731-42.

Role of the cyclic lipopeptide massetolide A in biological control of
Phytophthora infestans and in colonization of tomato plants by Pseudomonas
fluorescens.

Tran H, Ficke A, Asiimwe T, Hö¶fte M, Raaijmakers JM.

Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands.

Pseudomonas strains have shown promising results in biological control of late
blight caused by Phytophthora infestans. However, the mechanism(s) and
metabolites involved are in many cases poorly understood. Here, the role of the
cyclic lipopeptide massetolide A of Pseudomonas fluorescens SS101 in biocontrol
of tomato late blight was examined. Pseudomonas fluorescens SS101 was effective
in preventing infection of tomato (Lycopersicon esculentum) leaves by P.
infestans and significantly reduced the expansion of existing late blight
lesions. Massetolide A was an important component of the activity of P.
fluorescens SS101, since the massA-mutant was significantly less effective in
biocontrol, and purified massetolide A provided significant control of P.
infestans, both locally and systemically via induced resistance. Assays with nahG
transgenic plants indicated that the systemic resistance response induced by
SS101 or massetolide A was independent of salicylic acid signalling. Strain SS101
colonized the roots of tomato seedlings significantly better than its
massA-mutant, indicating that massetolide A was an important trait in plant
colonization. This study shows that the cyclic lipopeptide surfactant massetolide
A is a metabolite with versatile functions in the ecology of P. fluorescens SS101
and in interactions with tomato plants and the late blight pathogen P. infestans.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17688588 [PubMed - indexed for MEDLINE]

380: New Phytol. 2007;175(4):619-29.

Comment in:
New Phytol. 2007;175(4):597-9.

S cysteine-rich (SCR) binding domain analysis of the Brassica
self-incompatibility S-locus receptor kinase.

Kemp BP, Doughty J.

Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath
BA2 7AY, UK.

Brassica self-incompatibility, a highly discriminating outbreeding mechanism, has
become a paradigm for the study of plant cell-cell communications. When
self-pollen lands on a stigma, the male ligand S cysteine-rich (SCR), which is
present in the pollen coat, is transmitted to the female receptor, S-locus
receptor kinase (SRK). SRK is a membrane-spanning serine/threonine receptor
kinase present in the stigmatic papillar cell membrane. Haplotype-specific
binding of SCR to SRK brings about pollen rejection. The extracellular receptor
domain of SRK (eSRK) is responsible for binding SCR. Based on sequence homology,
eSRK can be divided into three subdomains: B lectin-like, hypervariable, and PAN.
Biochemical analysis of these subdomains showed that the hypervariable subdomain
is responsible for most of the SCR binding capacity of eSRK, whereas the B
lectin-like and PAN domains have little, if any, affinity for SCR. Fine mapping
of the SCR binding region of SRK using a peptide array revealed a region of the
hypervariable subdomain that plays a key role in binding the SCR molecule. We
show that residues within the hypervariable subdomain define SRK binding and are
likely to be involved in defining haplotype specificity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17688579 [PubMed - indexed for MEDLINE]

381: Mol Ecol. 2007 Aug;16(16):3292-8.

The temporal development in a hybridizing population of wild and cultivated
chicory (Cichorium intybus L.).

Sö¸rensen BS, Kiaer LP, Jö¸rgensen RB, Hauser TP.

Biosystems Department, Risö¸ National Laboratory, Frederiksborgvej 399, DK-4000
Roskilde, Denmark.

Hybridization and its possible impacts is a subject of increased attention in
connection with the risk of unintended gene flow from cultivated (including
genetically modified) plants to wild relatives. Whether such gene flow by
hybridization is likely to take place depends among other things on the
persistence of the hybrids in a natural environment over time. To evaluate this,
we studied an experimental hybridizing population of wild and cultivated
chicories (Cichorium intybus) relative to a previous study on the same population
2 years earlier. We compared the genetic composition, morphology and fitness
traits of plants from 2004 to the plants in the same plot in 2002. The majority
of the plants in 2004 was more morphologically and genetically intermediate than
in 2002. This indicates that no selection towards being wild-like or
cultivar-like was present over the period of 2 years. Furthermore, no distinct
fitness differences existed between the plants of 2004, probably due to most of
the plants being intermediate. No hybridization barriers appeared to be present
between wild and cultivated chicories beyond the F1 generation, since F2 hybrids
and backcrosses were in abundance; in fact, hybrids of probably fourth or fifth
generation were present. In conclusion, all results indicate that no barriers
exist to the temporal persistence of chicory hybrids in a natural environment.

PMID: 17688533 [PubMed - indexed for MEDLINE]

382: In Silico Biol. 2007;7(1):77-86.

Improved prediction of allergenicity by combination of multiple sequence motifs.

Kong W, Tan TS, Tham L, Choo KW.

Bioinformatics Group, Nanyang Polytechnic, Singapore. KONG_Wai_Ming@nyp.gov.sg

The identification and validation of protein allergens have become more important
nowadays as more and more transgenic proteins are introduced into our food
chains. Current allergen prediction algorithms focus on the identification of
single motif or single allergen peptide for allergen detection. However, an
analysis of the 575 allergen dataset shows that most allergens contain multiple
motifs. Here, we present a novel algorithm that detects allergen by making use of
combinations of motifs. Sensitivity of 0.772 and specificity of 0.904 were
achieved by the proposed algorithm to predict allergen. The specificity of the
proposed approach is found to be significantly higher than traditional single
motif approaches. The high specificity of the proposed algorithm is useful in
filtering out false positives, especially when laboratory resources are limited.

PMID: 17688432 [PubMed - indexed for MEDLINE]

383: Nat Biotechnol. 2007 Aug;25(8):874-5.

Comment on:
Nat Biotechnol. 2007 Aug;25(8):930-7.

The ABCs of low-phytate crops.

Raboy V.

Publication Types:
Comment
News

PMID: 17687363 [PubMed - indexed for MEDLINE]

384: Forum Nutr. 2007;60:183-95.

Prospects for improving the nutritional quality of dairy and meat products.

Coffey SG.

CSIRO Livestock Industries, St. Lucia, Australia. s.coffey@irl.cri.nz

Knowledge of the function of human and animal genes and their interactions is
rapidly increasing as a result of the completion of sequencing efforts for the
human, bovine and other genomes. Through transcriptomics, proteomics and
metabolomics, we have the capacity to study the health effects of food compounds
at the molecular level. The same tools that can assist the understanding of
nutrigenomics in humans can also be applied to producing animal-derived foods
with desired capacities to alter gene expression in humans. This, essentially,
represents food taking another major step in value through the personalisation of
health and nutrition. In its own right, nutrigenomics offers the potential to
improve animal production enterprises through major health and productivity
gains.

Publication Types:
Review

PMID: 17684415 [PubMed - indexed for MEDLINE]

385: BMC Neurosci. 2007 Aug 6;8:65.

Exploratory behaviour in NO-dependent cyclase mutants of Drosophila shows defects
in coincident neuronal signalling.

Tinette S, Zhang L, Garnier A, Engler G, Tares S, Robichon A.

Universitö© de Bourgogne, Dijon, France. tinette@cesg.cnrs.fr


BACKGROUND: Drosophila flies explore the environment very efficiently in order to
colonize it. They explore collectively, not individually, so that when a few land
on a food spot, they attract the others by signs. This behaviour leads to
aggregation of individuals and optimizes the screening of mates and egg-laying on
the most favourable food spots. RESULTS: Flies perform cycles of
exploration/aggregation depending on the resources of the environment. This
behavioural ecology constitutes an excellent model for analyzing simultaneous
processing of neurosensory information. We reasoned that the decision of flies to
land somewhere in order to achieve aggregation is based on simultaneous
integration of signals (visual, olfactory, acoustic) during their flight. On the
basis of what flies do in nature, we designed laboratory tests to analyze the
phenomenon of neuronal coincidence. We screened many mutants of genes involved in
neuronal metabolism and the synaptic machinery. CONCLUSION: Mutants of
NO-dependent cyclase show a specifically-marked behaviour phenotype, but on the
other hand they are associated with moderate biochemical defects. We show that
these mutants present errors in integrative and/or coincident processing of
signals, which are not reducible to the functions of the peripheral sensory
cells.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17683617 [PubMed - indexed for MEDLINE]

386: Plant Mol Biol. 2007 Oct;65(3):219-32. Epub 2007 Aug 8.

DORNROSCHEN-LIKE, an AP2 gene, is necessary for stamen emergence in Arabidopsis.

Nag A, Yang Y, Jack T.

Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.

To identify novel genes in petal and stamen development, a genetic screen was
carried out for enhancers of the unusual B class mutant pistillata-5 (pi-5). In
pi-5 flowers, second whorl organs develop as sepals rather than petals, but third
whorl stamens are normal. One pi-5 enhancer, dornrö¶schen-like-2 (drnl-2), results
in third whorl positions developing as filamentous organs. In addition to
enhancing the pi-5 phenotype, drnl-2 mutants also exhibit a phenotype in a
wild-type PI background. Although stamen primordia are morphologically visible
during early stages of flower development, they fail to enlarge in drnl-2
mutants. DRNL, which encodes a single AP2 domain protein, is expressed in a
dynamic pattern in the embryo, seedling, and flower. Analysis of both the drnl-2
mutant phenotype and the DRNL expression pattern in flowers suggests that DRNL
plays a critical role in stamen emergence in Arabidopsis.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17682829 [PubMed - indexed for MEDLINE]

387: Gastroenterology. 2007 Aug;133(2):517-28. Epub 2007 May 3.

Comment in:
Gastroenterology. 2007 Aug;133(2):706-9.

Induction of ovalbumin-specific tolerance by oral administration of Lactococcus
lactis secreting ovalbumin.

Huibregtse IL, Snoeck V, de Creus A, Braat H, De Jong EC, Van Deventer SJ,
Rottiers P.

Center for Experimental and Molecular Medicine, Academic Medical Center,
University of Amsterdam, Amsterdam, The Netherlands.

BACKGROUND AND AIMS: Obtaining antigen-specific immune suppression is an
important goal in developing treatments of autoimmune, inflammatory, and allergic
gastrointestinal diseases. Oral tolerance is a powerful means for inducing
tolerance to a particular antigen, but implementing this strategy in humans has
been difficult. Active delivery of recombinant autoantigens or allergens at the
intestinal mucosa by genetically modified Lactococcus lactis (L lactis) provides
a novel therapeutic approach for inducing tolerance. METHODS: We engineered the
food grade bacterium L lactis to secrete ovalbumin (OVA) and evaluated its
ability to induce OVA-specific tolerance in OVA T-cell receptor (TCR) transgenic
mice (DO11.10). Tolerance induction was assessed by analysis of delayed-type
hypersensitivity responses, measurement of cytokines and OVA-specific
proliferation, phenotypic analysis, and adoptive transfer experiments. RESULTS:
Intragastric administration of OVA-secreting L lactis led to active delivery of
OVA at the mucosa and suppression of local and systemic OVA-specific T-cell
responses in DO11.10 mice. This suppression was mediated by induction of
CD4(+)CD25(-) regulatory T cells that function through a transforming growth
factor beta-dependent mechanism. Restimulation of splenocytes and gut-associated
lymph node tissue from these mice resulted in a significant OVA-specific decrease
in interferon gamma and a significant increase in interleukin-10 production.
Furthermore, Foxp3 and CTLA-4 were significantly up-regulated in the
CD4(+)CD25(-) population. CONCLUSIONS: Mucosal antigen delivery by oral
administration of genetically engineered L lactis leads to antigen-specific
tolerance. This approach can be used to develop effective therapeutics for
systemic and intestinal immune-mediated inflammatory diseases.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17681173 [PubMed - indexed for MEDLINE]

388: Plant Cell Rep. 2007 Dec;26(12):2111-7. Epub 2007 Aug 7.

Yeast-based recombineering of DNA fragments into plant transformation vectors by
one-step transformation.

Nagano Y, Takao S, Kudo T, Iizasa E, Anai T.

Analytical Research Center for Experimental Sciences, Saga University, Saga,
Japan. nagano@cc.saga-u.ac.jp

T-DNA binary vectors are often used in plant transformation experiments. Because
they are usually very large and have few restriction sites suitable for DNA
ligation reactions, cloning DNA fragments into these vectors is difficult. We
provide herein an alternative to cloning DNA fragments into very large vectors.
Our yeast-based recombineering method enables DNA fragments to be cloned into
certain types of T-DNA binary vectors by one-step transformation without the
requirement of specific recombination sites or precisely positioned restriction
ends, thus making the cloning process more flexible. Moreover, this method is
inexpensive and is applicable to multifragment cloning.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17680244 [PubMed - indexed for MEDLINE]

389: Duke Law J. 2007 Apr;56(6):1581-6.

Beyond food and evil.

Chen J.

University of Louisville, Louis D. Brandeis School of Law.

PMID: 17679180 [PubMed - indexed for MEDLINE]

390: Dev Genes Evol. 2007 Sep;217(9):629-37. Epub 2007 Aug 3.

A rice YABBY gene, OsYABBY4, preferentially expresses in developing vascular
tissue.

Liu HL, Xu YY, Xu ZH, Chong K.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, the Chinese Academy of Sciences, Nanxincun 20, Xiangshan,
Beijing, 100093, People's Republic of China.

Developmental gene families have diversified during land plant evolution. The
primary role of YABBY gene family is promoting abaxial fate in model eudicot,
Arabidopsis thaliana. However recent results suggest that roles of YABBY genes
are not conserved in the angiosperms. In this paper, a rice YABBY gene was
isolated, and its expression patterns were analyzed in detail. Sequence
characterization and phylogenetic analyses showed the gene is OsYABBY4, which is
group-classified into FIL/YAB3 subfamily. Beta-glucuronidase reporter assay and
in situ analysis consistently revealed that OsYABBY4 was expressed in the
meristems and developing vascular tissue of rice, predominantly in the phloem
tissue, suggesting that the function of the rice gene is different from those of
its counterparts in eudicots. OsYABBY4 may have been recruited to regulate the
development of vasculature in rice. However, transgenic Arabidopsis plants
ectopically expressing OsYABBY4 behaved very like those over-expressing FIL or
YAB3 with abaxialized lateral organs, suggesting the OsYABBY4 protein domain is
conserved with its Arabidopsis counterparts in sequences. Our results also
indicate that the functional diversification of OsYABBY4 may be associated with
the divergent spatial-temporal expression patterns, and YABBY family members may
have preserved different expression regulatory systems and functions during the
evolution of different kinds of species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17676337 [PubMed - indexed for MEDLINE]

391: Nat Biotechnol. 2007 Aug;25(8):930-7. Epub 2007 Aug 5.

Comment in:
Nat Biotechnol. 2007 Aug;25(8):874-5.

Embryo-specific silencing of a transporter reduces phytic acid content of maize
and soybean seeds.

Shi J, Wang H, Schellin K, Li B, Faller M, Stoop JM, Meeley RB, Ertl DS, Ranch
JP, Glassman K.

Crop Genetics Research and Development, Pioneer Hi-Bred International, A DuPont
Company, Johnston, Iowa 50131, USA. Jinrui.shi@Pioneer.com

Phytic acid in cereal grains and oilseeds is poorly digested by monogastric
animals and negatively affects animal nutrition and the environment. However,
breeding programs involving mutants with less phytic acid and more inorganic
phosphate (P(i)) have been frustrated by undesirable agronomic characteristics
associated with the phytic acid-reducing mutations. We show that maize lpa1
mutants are defective in a multidrug resistance-associated protein (MRP)
ATP-binding cassette (ABC) transporter that is expressed most highly in embryos,
but also in immature endosperm, germinating seed and vegetative tissues.
Silencing expression of this transporter in an embryo-specific manner produced
low-phytic-acid, high-Pi transgenic maize seeds that germinate normally and do
not show any significant reduction in seed dry weight. This dominant transgenic
approach obviates the need for incorporating recessive lpa1 mutations to create
maize hybrids with reduced phytic acid. Suppressing the homologous soybean MRP
gene also generated low-phytic-acid seed, suggesting that the strategy might be
feasible for many crops.

PMID: 17676037 [PubMed - indexed for MEDLINE]

392: Plant Cell Physiol. 2007 Sep;48(9):1331-9. Epub 2007 Aug 3.

Ectopic expression of a foreign aquaporin disrupts the natural expression
patterns of endogenous aquaporin genes and alters plant responses to different
stress conditions.

Jang JY, Rhee JY, Kim DG, Chung GC, Lee JH, Kang H.

Department of Plant Biotechnology, Agricultural Plant Stress Research Center and
Biotechnology Research Institute, College of Agriculture and Life Sciences,
Chonnam National University, Buk-Gu, Gwangju, Korea.

Although the number of reports demonstrating the roles of individual aquaporins
in plants under diverse physiological conditions is expanding, the importance of
interactions between different aquaporin isoforms and their integrated functions
under stress conditions remain unclear. Here, we expressed one cucumber aquaporin
gene, designated CsPIP1;1, and one figleaf gourd aquaporin gene, designated
CfPIP2;1, in Arabidopsis thaliana, and investigated the effect of its expression
on the natural expression patterns of endogenous PIP genes under stress
conditions. The transcript levels of endogenous Arabidopsis PIP members were
altered differently depending on stress conditions by the expression of CsPIP1;1
or CfPIP2;1. The transgenic Arabidopsis plants that constitutively express
CfPIP2;1 displayed better growth compared with the wild-type plants under
dehydration stress conditions, whereas CsPIP1;1 expression exerted a negative
effect on the growth of Arabidopsis under dehydration stress conditions. CsPIP1;1
or CfPIP2;1 expression facilitated seed germination under high salt stress
conditions, but had no influence on the growth of Arabidopsis under cold stress
conditions. Our results indicate that the ectopic expression of a foreign
aquaporin gene perturbs differently the natural expression patterns of endogenous
aquaporin genes depending on particular stress conditions, and thereby influences
the responses of plants to different stress conditions. This implies that the up-
and/or down-regulation of aquaporins and their integrated functions are crucial
to the maintenance of proper water balance under stress conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17675323 [PubMed - indexed for MEDLINE]

393: Plant Cell Rep. 2007 Dec;26(12):2091-9. Epub 2007 Aug 1.

Expression and functional analysis of ZmDWF4, an ortholog of Arabidopsis DWF4
from maize (Zea mays L.).

Liu T, Zhang J, Wang M, Wang Z, Li G, Qu L, Wang G.

State Key Laboratory of Agrobiotechnology and National Center for Plant Gene
Research, China Agricultural University, Beijing, China.

DWF4 encodes a rate-limiting mono-oxygenase that mediates 22alpha-hydroxylation
reactions in the BR biosynthetic pathway and it is the target gene in the BR
feedback loop. Knockout of DWF4 results in a dwarfed phenotype and other severe
defects in Arabidopsis. Here we report on the isolation of the ZmDWF4 gene in
maize. Sequence analysis revealed that the open reading frame of ZmDWF4 was 1,518
bp, which encodes a protein composed of 505 amino acid residues with a calculated
molecular mass of 57.6 kD and a predicated isoelectric point (pI) of 9.54.
Phylogenetic analysis indicated that ZmDWF4 was very close to the Arabidopsis
DWF4. In young maize seedlings, the expression of ZmDWF4 in shoots was much
higher than that in roots. The highest expression of ZmDWF4 was observed in husk
leaves and the lowest in silks during flowering stage. The expression of ZmDWF4
in maize was significantly down regulated by exogenous brassinolide. A
heterogeneous complementary experiment demonstrated that the defects of three
Arabidopsis DWF4 mutants could be rescued by constitutive expression of ZmDWF4,
with leaf expandability, inflorescence stem heights and fertile capabilities all
restored to normal levels. Increases in seed and branch number as well as the
height of florescence stem were observed in the over-expressed transformants.
These findings suggest that ZmDWF4 may be an ortholog gene of Arabidopsis DWF4
and responsible for BR biosynthesis in maize.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17668219 [PubMed - indexed for MEDLINE]

394: Plant Cell Rep. 2007 Nov;26(11):1919-31. Epub 2007 Jul 28.

Promoters of two anther-specific genes confer organ-specific gene expression in a
stage-specific manner in transgenic systems.

Gupta V, Khurana R, Tyagi AK.

Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular
Biology, University of Delhi South Campus, New Delhi 110021, India.

Differential screening of a stage-specific cDNA library of Indica rice has been
used to identify two genes expressed in pre-pollination stage panicles, namely
OSIPA and OSIPK coding for proteins similar to expansins/pollen allergens and
calcium-dependent protein kinases (CDPK), respectively. Northern analysis and in
situ hybridizations indicate that OSIPA expresses exclusively in pollen while
OSIPK expresses in pollen as well as anther wall. Promoters of these two
anther-specific genes show the presence of various cis-acting elements (GTGA and
AGAAA) known to confer anther/pollen-specific gene expression.
Organ/tissue-specific activity and strength of their regulatory regions have been
determined in transgenic systems, i.e., tobacco and Arabidopsis. A unique
temporal activity of these two promoters was observed during various
developmental stages of anther/pollen. Promoter of OSIPA is active during the
late stages of pollen development and remains active till the anthesis, whereas,
OSIPK promoter is active to a low level in developing anther till the pollen
matures. OSIPK promoter activity diminishes before anthesis. Both promoters show
a potential to target expression of the gene of interest in developmental
stage-specific manner and can help engineer pollen-specific traits like
male-sterility in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17661051 [PubMed - indexed for MEDLINE]

395: Plant Cell. 2007 Jul;19(7):2124-39. Epub 2007 Jul 27.

Erratum in:
Plant Cell. 2007 Aug;19(8):2693-4.

Light-independent phytochrome signaling mediated by dominant GAF domain tyrosine
mutants of Arabidopsis phytochromes in transgenic plants.

Su YS, Lagarias JC.

Section of Molecular and Cellular Biology, University of California, Davis,
California 95616, USA.

The photoreversibility of plant phytochromes enables continuous surveillance of
the ambient light environment. Through expression of profluorescent,
photoinsensitive Tyr-to-His mutant alleles of Arabidopsis thaliana phytochrome B
(PHYB(Y276H)) and Arabidopsis phytochrome A (PHYA(Y242H)) in transgenic
Arabidopsis plants, we demonstrate that photoconversion is not a prerequisite for
phytochrome signaling. PHYB(Y276H)-expressing plants exhibit
chromophore-dependent constitutive photomorphogenesis, light-independent
phyB(Y276H) nuclear localization, constitutive activation of genes normally
repressed in darkness, and light-insensitive seed germination. Fluence rate
analyses of transgenic plants expressing PHYB(Y276H), PHYA(Y242H), and other
Y(GAF) mutant alleles of PHYB demonstrate that a range of altered light-signaling
activities are associated with mutation of this residue. We conclude that the
universally conserved GAF domain Tyr residue, with which the bilin chromophore is
intimately associated, performs a critical role in coupling light perception to
signal transduction by plant phytochromes.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17660358 [PubMed - indexed for MEDLINE]

396: Proc Natl Acad Sci U S A. 2007 Aug 21;104(34):13603-8. Epub 2007 Jul 27.

Comment in:
Proc Natl Acad Sci U S A. 2007 Aug 28;104(35):13859-60.

Recombinant human butyrylcholinesterase from milk of transgenic animals to
protect against organophosphate poisoning.

Huang YJ, Huang Y, Baldassarre H, Wang B, Lazaris A, Leduc M, Bilodeau AS,
Bellemare A, Cö´tö© M, Herskovits P, Touati M, Turcotte C, Valeanu L, Lemö©e N,
Wilgus H, Bö©gin I, Bhatia B, Rao K, Neveu N, Brochu E, Pierson J, Hockley DK,
Cerasoli DM, Lenz DE, Karatzas CN, Langermann S.

PharmAthene Canada, Inc., 7150 Alexander-Fleming, Montreal, QC, Canada H4S 2C8.
yjhuang@pharmathene.ca

Dangerous organophosphorus (OP) compounds have been used as insecticides in
agriculture and in chemical warfare. Because exposure to OP could create a danger
for humans in the future, butyrylcholinesterase (BChE) has been developed for
prophylaxis to these chemicals. Because it is impractical to obtain sufficient
quantities of plasma BChE to treat humans exposed to OP agents, the production of
recombinant BChE (rBChE) in milk of transgenic animals was investigated.
Transgenic mice and goats were generated with human BChE cDNA under control of
the goat beta-casein promoter. Milk from transgenic animals contained 0.1-5
g/liter of active rBChE. The plasma half-life of PEGylated, goat-derived,
purified rBChE in guinea pigs was 7-fold longer than non-PEGylated dimers. The
rBChE from transgenic mice was inhibited by nerve agents at a 1:1 molar ratio.
Transgenic goats produced active rBChE in milk sufficient for prophylaxis of
humans at risk for exposure to OP agents.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17660298 [PubMed - indexed for MEDLINE]

397: Trends Biotechnol. 2007 Sep;25(9):376-84. Epub 2007 Jul 30.

Humanizing infant milk formula to decrease postnatal HIV transmission.

Blais DR, Altosaar I.

Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine,
University of Ottawa, Ottawa, Ontario, K1H 8M5, Canada.

There are currently no safe methods for feeding babies born from the 16 million
HIV-infected women living in resource-constrained countries. Breast milk can
transmit HIV, and formula feeding can lead to gastrointestinal illnesses owing to
unsanitary conditions and the composition of milk formulations. There is
therefore a need to ensure that breast milk substitutes provide optimal health
outcomes. Given that the immune properties of several breast milk proteins are
known, transgenic food crops could facilitate inexpensive and safe reconstitution
of the beneficial breast milk proteome in infant formulae, while keeping the HIV
virus at bay. At least seven breast milk immune proteins have already been
produced in food crops, and dozens more proteins could potentially be produced if
fortified formula proves effective in nursing newborns born to HIV-infected
mothers.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17659799 [PubMed - indexed for MEDLINE]

398: Neuroscience. 2007 Aug 24;148(2):371-4. Epub 2007 Jul 19.

Co-regulation of cold-resistant food acquisition by insulin- and neuropeptide
Y-like systems in Drosophila melanogaster.

Lingo PR, Zhao Z, Shen P.

Department of Cellular Biology, and Biomedical and Health Sciences Institute, 724
Biological Sciences Building, University of Georgia, Athens, GA 30602, USA.

To survive, food-deprived animals may be forced to forage under hostile
conditions. We attempt to use genetically tractable Drosophila melanogaster as a
model to elucidate molecular and neural mechanisms that drive a forager to engage
in risk-prone food acquisition. Here we describe a paradigm for assessing
hunger-driven food acquisition by fly larvae at a deleteriously cold temperature.
Genetic analyses reveal that the neural activity of NPFR1, a receptor of
neuropeptide F (NPF, the sole fly homolog of neuropeptide Y or NPY), was required
for cold-resistant feeding behavior of fasted larvae. Conversely, NPFR1
overexpression in fed larvae was sufficient to trigger cold-resistant feeding
activity normally associated with fasted larvae. Furthermore, the fly
insulin-like system, implicated in the transduction of hunger signals to the CNS,
regulated negatively larval cold-resistant food acquisition. The results from
this and our previous studies suggest that the fly NPY-like system is a central
mediator of hunger-elicited resistance to diverse stressors that can be of
thermal, gustatory or mechanical form.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 17658221 [PubMed - indexed for MEDLINE]

399: Food Nutr Bull. 2007 Jun;28(2 Suppl):S271-9.

From harvest to health: challenges for developing biofortified staple foods and
determining their impact on micronutrient status.

Hotz C, McClafferty B.

HarvestPlus in Washington, DC 20006-1002, USA.

BACKGROUND: The use of conventional breeding techniques and biotechnology to
improve the micronutrient quality of staple crops is a new strategy to address
micronutrient deficiencies in developing countries. This strategy, referred to as
"biofortification," is being developed and implemented through the international
alliance of HarvestPlus to improve iron, zinc, and vitamin A status in low-income
populations. OBJECTIVE: The objective of this paper is to review the challenges
faced by nutritionists to determine and demonstrate the ability of biofortified
crops to have an impact on the nutritional and health status of target
populations. METHODS: We reviewed available published and unpublished information
that is needed to design and evaluate this strategy, including issues related to
micronutrient retention in staple foods, micronutrient bioavailability from plant
foods, and evidence for the efficacy of high-micronutrient-content staple foods
to improve micronutrient status. RESULTS: Further information is needed on the
retention of micronutrients in staple foods, in particular of provitamin A
carotenoids, when stored and prepared under different conditions. The low
bioavailability of iron from staple foods and the ability to demonstrate an
impact on zinc status are specific challenges that need to be addressed. In
target countries, infections and other micronutrient deficiencies may confound
the ability to affect micronutrient status, and this must be taken into account
in community-based studies. CONCLUSIONS: Information to date suggests that
biofortification has the potential to contribute to increased micronutrient
intakes and improved micronutrient status. The success of this strategy will
require the collaboration between health and agriculture sectors.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17658073 [PubMed - indexed for MEDLINE]

400: Food Nutr Bull. 2007 Jun;28(2 Suppl):S258-70.

Ensuring the supply of and creating demand for a biofortified crop with a visible
trait: lessons learned from the introduction of orange-fleshed sweet potato in
drought-prone areas of Mozambique.

Low JW, Arimond M, Osman N, Cunguara B, Zano F, Tschirley D.

Jan W Low was affiliated with the Department of Agricultural Economics, Michigan
State University, East Lansing, Michigan, USA. j.low@cgiar.org

BACKGROUND: Orange-fleshed sweet potato (OFSP) is a promising biofortified crop
for sub-Saharan Africa because it has high levels of provitamin A carotenoids,
the formed vitamin A is bioavailable, and white-fleshed sweet potato is already
widely grown. OBJECTIVES: To examine whether farmers will adopt varieties with a
distinct visible trait, young children will eat OFSP in sufficient quantities to
improve vitamin A intake, OFSP can serve as an entry point for promoting a more
diversified diet, and lessons can be drawn to assure sustained adoption. METHODS:
The 2-year quasi-experimental intervention study followed households and children
(n = 741; mean age, 13 months at baseline) through two agricultural cycles in
drought prone-areas of Mozambique. RESULTS: OFSP is acceptable to farmers when
introduced by using an integrated approach. In the second year, intervention
children (n = 498) were more likely than control children (n = 243) to have
consumed OFSP (54% vs. 4%), dark-green leaves (60% vs. 46%), or ripe papaya (65%
vs. 42%) on 3 or more days in the previous week (p < .001 for all comparisons).
Their vitamin A intakes were nearly eight times higher than those of control
children (median, 426 vs. 56 1g RAE [retinol activity equivalents], p < .001).
Diet diversification was limited by difficult agroecological conditions and low
purchasing power. However, dietary diversity was higher among intervention than
control children (32% vs. 9% consuming food from more than four groups; p <
.001). CONCLUSIONS: An integrated OFSP-based approach had a positive impact on
the vitamin A intake of young children. A market development component and
improved vine multiplication systems are recommended to assure sustained
adoption.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17658072 [PubMed - indexed for MEDLINE]

401: Shokuhin Eiseigaku Zasshi. 2007 Jun;48(3):41-50.

[A 52-week feeding study of genetically modified soybeans in F344 rats]

[Article in Japanese]

Sakamoto Y, Tada Y, Fukumori N, Tayama K, Ando H, Takahashi H, Kubo Y, Nagasawa
A, Yano N, Yuzawa K, Ogata A, Kamimura H.

Department of Environmental Health and Toxicology, Tokyo Metropolitan Institute
of Public Health: 3-24-1 Hyakunin-cho, Shinjuku-ku, Tokyo 169-0073, Japan.

A chronic feeding study to evaluate the safety of the genetically modified
glyphosate-tolerant soybeans (GM soybeans) was conducted using rats. F344 DuCrj
rats were fed diet containing GM soybeans or Non-GM soybeans at the concentration
of 30% in basal diet. Non-GM soybeans were closely related strain of GM soybeans.
These two diets were adjusted to an identical nutrient level. In this study, the
influence of GM soybeans on rats was compared with that of the Non-GM soybeans,
and furthermore, to assess the effect of soybeans themselves, the groups of rats
fed GM and Non-GM soybeans were compared with a group fed commercial diet (CE-2).
General conditions were observed daily and body weight and food consumption were
recorded. At the intermediate examination (26 weeks), and at the termination (52
weeks), animals were subjected to hematology, serum biochemistry, and
pathological examination. There were several differences in animal growth, food
intake, serum biochemical parameters and histological findings between the rats
fed the GM and/or Non-GM soybeans and the rats fed CE-2. However, body weight and
food intake were similar for the rats fed the GM and Non-GM soybeans. Gross
necropsy findings, hematological and serum biochemical parameters, organ weights,
and pathological findings showed no meaningful difference between rats fed the GM
and Non-GM soybeans. These results indicate that long-term intake of GM soybeans
at the level of 30% in diet has no apparent adverse effect in rats.

Publication Types:
English Abstract

PMID: 17657996 [PubMed - indexed for MEDLINE]

402: Theor Appl Genet. 2007 Nov;115(7):887-95. Epub 2007 Jul 27.

Functional analysis of Xa3/Xa26 family members in rice resistance to Xanthomonas
oryzae pv. oryzae.

Cao Y, Duan L, Li H, Sun X, Zhao Y, Xu C, Li X, Wang S.

National Key Laboratory of Crop Genetic Improvement, National Center of Plant
Gene Research (Wuhan), Huazhong Agricultural University, Wuhan 430070, China.

Plant disease resistant (R) genes are frequently clustered in the genome. The
diversity of members in a complex R-gene family may provide variation in
resistance specificity. Rice Xa3/Xa26, conferring resistance to Xanthomonas
oryzae pv. oryzae (Xoo) encodes a leucine-rich repeat (LRR) receptor kinase-type
protein and belongs to a multigene family, consisting of Xa3/Xa26, MRKa, MRKc and
MRKd in rice cultivar Minghui 63. MRKa and MRKc are intact genes, while MRKd is a
pseudogene. Complementary analyses showed that MRKa and MRKc could not mediate
resistance to Xoo when regulated by their native promoters, but MRKa not MRKc
conferred partial resistance to Xoo when regulated by a strong constitutive
promoter. Plants carrying truncated XA3/XA26, which lacked the kinase domain,
were compromised in their resistance to Xoo. However, the kinase domain of MRKa
could partially restore the function of the truncated XA3/XA26 in resistance.
MRKa and MRKc showed similar expression pattern as Xa3/Xa26, which expressed only
in the vascular systems of different tissues. The expressional characteristic of
MRKa and MRKc perfectly fits the function of genes conferring resistance to Xoo,
a vascular pathogen. These results suggest that although MRKa and MRKc cannot
mediate bacterial blight resistance nowadays, they may be once effective genes
for Xoo resistance. Their expressional characteristic and sequence similarity to
Xa3/Xa26 will provide templates for generating novel recognition specificity to
face the evolution of Xoo. In addition, both LRR and kinase domains encoded by
Xa3/Xa26 and MRKa are the functional determinants and MRKa-mediated resistance is
dosage-dependent.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17657469 [PubMed - indexed for MEDLINE]

403: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 12;363(1491):591-609.

Genetic contributions to agricultural sustainability.

Dennis ES, Ellis J, Green A, Llewellyn D, Morell M, Tabe L, Peacock WJ.

CSIRO Plant Industry, GPO Box 1600, Canberra, Australian Capital Territory 2601,
Australia.

The current tools of enquiry into the structure and operation of the plant genome
have provided us with an understanding of plant development and function far
beyond the state of knowledge that we had previously. We know about key genetic
controls repressing or stimulating the cascades of gene expression that move a
plant through stages in its life cycle, facilitating the morphogenesis of
vegetative and reproductive tissues and organs. The new technologies are enabling
the identification of key gene activity responses to the range of biotic and
abiotic challenges experienced by plants. In the past, plant breeders produced
new varieties with changes in the phases of development, modifications of plant
architecture and improved levels of tolerance and resistance to environmental and
biotic challenges by identifying the required phenotypes in a few plants among
the large numbers of plants in a breeding population. Now our increased knowledge
and powerful gene sequence-based diagnostics provide plant breeders with more
precise selection objectives and assays to operate in rationally planned crop
improvement programmes. We can expect yield potential to increase and harvested
product quality portfolios to better fit an increasing diversity of market
requirements. The new genetics will connect agriculture to sectors beyond the
food, feed and fibre industries; agri-business will contribute to public health
and will provide high-value products to the pharmaceutical industry as well as to
industries previously based on petroleum feedstocks and chemical modification
processes.

Publication Types:
Review

PMID: 17656342 [PubMed - indexed for MEDLINE]

404: Plant Cell Rep. 2007 Dec;26(12):2071-82. Epub 2007 Jul 26.

Stress-inducible expression of At DREB1A in transgenic peanut (Arachis hypogaea
L.) increases transpiration efficiency under water-limiting conditions.

Bhatnagar-Mathur P, Devi MJ, Reddy DS, Lavanya M, Vadez V, Serraj R,
Yamaguchi-Shinozaki K, Sharma KK.

International Crops Research Institute for the Semi-Arid Tropics (ICRISAT),
Patancheru, AP, India.

Water deficit is the major abiotic constraint affecting crop productivity in
peanut (Arachis hypogaea L.). Water use efficiency under drought conditions is
thought to be one of the most promising traits to improve and stabilize crop
yields under intermittent water deficit. A transcription factor DREB1A from
Arabidopsis thaliana, driven by the stress inducible promoter from the rd29A
gene, was introduced in a drought-sensitive peanut cultivar JL 24 through
Agrobacterium tumefaciens-mediated gene transfer. The stress inducible expression
of DREB1A in these transgenic plants did not result in growth retardation or
visible phenotypic alterations. T3 progeny of fourteen transgenic events were
exposed to progressive soil drying in pot culture. The soil moisture threshold
where their transpiration rate begins to decline relative to control well-watered
(WW) plants and the number of days needed to deplete the soil water was used to
rank the genotypes using the average linkage cluster analysis. Five diverse
events were selected from the different clusters and further tested. All the
selected transgenic events were able to maintain a transpiration rate equivalent
to the WW control in soils dry enough to reduce transpiration rate in wild type
JL 24. All transgenic events except one achieved higher transpiration efficiency
(TE) under WW conditions and this appeared to be explained by a lower stomatal
conductance. Under water limiting conditions, one of the selected transgenic
events showed 40% higher TE than the untransformed control.

PMID: 17653723 [PubMed - indexed for MEDLINE]

405: Mol Biotechnol. 2007 Mar;35(3):215-23.

Expression of a synthetic neutralizing epitope of porcine epidemic diarrhea virus
fused with synthetic B subunit of Escherichia coli heat labile enterotoxin in
rice endosperm.

Oszvald M, Kang TJ, Tomoskozi S, Tamas C, Tamas L, Kim TG, Yang MS.

Department of Biochemistry and Food Technology, Budapest University of Technology
and Economics, Budapest, Hungary.

Epitopes often require co-delivery with adjuvant and targeting proteins to enable
recognition by the immune system, and this approach may also increase the
efficacy of the antigen. In this study, we assess and describe the ability of
transgenic rice plants to express a fusion protein consisting of the B-subunit of
the Escherichia coli heat-labile enterotoxin (LTB) and a synthetic
core-neutralizing epitope (COE) of porcine epidemic diarrhea virus (PEDV),
inducing an enteric disease that is seen most predominantly in piglets. Both
components of the fusion proteins were detected with Western blot analysis. The
fusion protein was determined to assemble into pentamers, as was evidenced by its
ability to bind to GM1 gangliosides, and evidenced an average level of expression
in a transgenic rice endosperm. This indicates that the expression system of the
plant is capable of generating a sizable amount of antigen, possibly allowing for
the successful development of an edible vaccine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17652785 [PubMed - indexed for MEDLINE]

406: Development. 2007 Sep;134(17):3099-109. Epub 2007 Jul 25.

Haploinsufficiency after successive loss of signaling reveals a role for
ERECTA-family genes in Arabidopsis ovule development.

Pillitteri LJ, Bemis SM, Shpak ED, Torii KU.

Department of Biology, University of Washington, Seattle, WA 98195, USA.

The Arabidopsis genome contains three ERECTA-family genes, ERECTA (ER),
ERECTA-LIKE 1 (ERL1) and ERL2 that encode leucine-rich repeat receptor-like
kinases. This gene family acts synergistically to coordinate cell proliferation
and growth during above-ground organogenesis with the major player, ER, masking
the loss-of-function phenotypes of the other two members. To uncover the specific
developmental consequence and minimum threshold requirement for signaling,
ER-family gene function was successively eliminated. We report here that ERL2 is
haploinsufficient for maintaining female fertility in the absence of ER and ERL1.
Ovules of the haploinsufficient er-105 erl1-2 erl2-1/+ mutant exhibit abnormal
development with reduced cell proliferation in the integuments and gametophyte
abortion. Our analysis indicates that progression of integument growth requires
ER-family signaling in a dosage-dependent manner and that transcriptional
compensation among ER-family members occurs to maintain the required signaling
threshold. The specific misregulation of cyclin A genes in the er-105 erl1-2
erl2-1/+ mutant suggests that downstream targets of the ER-signaling pathway
might include these core cell-cycle regulators. Finally, genetic interaction of
the ER family and the WOX-family gene, PFS2, reveals their contribution to
integument development through interrelated mechanisms.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17652352 [PubMed - indexed for MEDLINE]

407: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 12;363(1491):611-21.

Integrated pest management: the push-pull approach for controlling insect pests
and weeds of cereals, and its potential for other agricultural systems including
animal husbandry.

Hassanali A, Herren H, Khan ZR, Pickett JA, Woodcock CM.

International Centre of Insect Physiology and Ecology, PO Box 30772, Nairobi,
Kenya.

This paper describes the 'push-pull' or 'stimulo-deterrent diversionary' strategy
in relation to current and potential examples from our own experiences. The
push-pull effect is established by exploiting semiochemicals to repel insect
pests from the crop ('push') and to attract them into trap crops ('pull'). The
systems exemplified here have been developed for subsistence farming in Africa
and delivery of the semiochemicals is entirely by companion cropping, i.e.
intercropping for the push and trap cropping for the pull. The main target was a
series of lepidopterous pests attacking maize and other cereals. Although the
area given to the cereal crop itself is reduced under the push-pull system,
higher yields are produced per unit area. An important spin-off from the project
is that the companion crops are valuable forage for farm animals. Leguminous
intercrops also provide advantages with regard to plant nutrition and some of the
trap crops help with water retention and in reducing land erosion. A major
benefit is that certain intercrop plants provide dramatic control of the African
witchweed (striga). Animal husbandry forms an essential part of intensive
subsistence agriculture in Africa and developments using analogous push-pull
control strategies for insect pests of cattle are exemplified.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17652071 [PubMed - indexed for MEDLINE]

408: Philos Trans R Soc Lond B Biol Sci. 2008 Feb 12;363(1491):639-58.

Improving water use in crop production.

Morison JI, Baker NR, Mullineaux PM, Davies WJ.

Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester
CO4 3SQ, UK. morisj@essex.ac.uk

Globally, agriculture accounts for 80-90% of all freshwater used by humans, and
most of that is in crop production. In many areas, this water use is
unsustainable; water supplies are also under pressure from other users and are
being affected by climate change. Much effort is being made to reduce water use
by crops and produce 'more crop per drop'. This paper examines water use by
crops, taking particularly a physiological viewpoint, examining the underlying
relationships between carbon uptake, growth and water loss. Key examples of
recent progress in both assessing and improving crop water productivity are
described. It is clear that improvements in both agronomic and physiological
understanding have led to recent increases in water productivity in some crops.
We believe that there is substantial potential for further improvements owing to
the progress in understanding the physiological responses of plants to water
supply, and there is considerable promise within the latest molecular genetic
approaches, if linked to the appropriate environmental physiology. We conclude
that the interactions between plant and environment require a team approach
looking across the disciplines from genes to plants to crops in their particular
environments to deliver improved water productivity and contribute to
sustainability.

Publication Types:
Review

PMID: 17652070 [PubMed - indexed for MEDLINE]

409: Ying Yong Sheng Tai Xue Bao. 2007 May;18(5):1077-80.

[Effects of transgenic corn expressing Bacillus thuringiensis cry1Ab toxin on
population increase of Rhopalosiphum maidis Fitch]

[Article in Chinese]

Li LL, Wang ZY, He KL, Bai SX, Hua L.

State Key Laboratory for the Biology of Plant Diseases and Insect Pests,
Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing
100094, China. wl3pq@sohu.com

A laboratory study was made on the effects of transgenic corn expressing cry1Ab
toxin from the bacterium Bacillus thuringiensis (Bt) on the life table of an
experimental population of corn leaf aphid Rhopalosiphum maidis Fitch. The
results showed that no obvious negative effects of two Bt corn hybrids DK647BTY
(event MON810) and NX4777 (event Bt11) were observed on the net reproductive
rate, average generation time, innate capacity for increase, and finite rate of
increase of R. maidis, and there were no significant differences in the rate of
alate, mortality of nymph, and longevity and fecundity duration of corn leaf
aphid when feeding on Bt and non-Bt corn hybrids, suggesting that Bt corn
expressing cry1Ab toxin had no side-effects on the development and reproduction
of R. maidis.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 17650861 [PubMed - indexed for MEDLINE]

410: Ann Bot (Lond). 2007 Sep;100(3):651-7. Epub 2007 Jul 23.

Functional conservation between CRABS CLAW orthologues from widely diverged
angiosperms.

Fourquin C, Vinauger-Douard M, Chambrier P, Berne-Dedieu A, Scutt CP.

Laboratoire de Reproduction et Dö©veloppement des Plantes, Ecole Normale
Supö©rieure de Lyon, 46, allö©e d'Italie, 69364 Lyon Cedex 07, France.

BACKGROUND AND AIMS: CRABS CLAW (CRC) encodes a transcription factor of the YABBY
family that plays important roles in carpel and nectary development in
Arabidopsis thaliana. Combined evolutionary and developmental studies suggest an
ancestor of the CRC gene to have controlled carpel development in the last common
ancestor of the angiosperms. Roles for CRC orthologues in leaf development and
carpel specification in rice, and in nectary development in core eudicots, have
accordingly been interpreted as derived. The aim of this study was to assess the
capacity of CRC orthologues from a basal angiosperm and from rice to complement
CRC mutants of arabidopsis. These experiments were designed to test the
hypothesized ancestral role of CRC in the angiosperms, and to indicate whether
putatively novel roles of various CRC orthologues resulted from changes to their
encoded proteins, or from other molecular evolutionary events. METHODS: The crc-1
mutant of arabidopsis was genetically transformed with the coding sequences of
various CRC orthologues, and with paralogous YABBY coding sequences, under the
control of the arabidopsis CRC promoter. The phenotypes of transformed plants
were assessed to determine the degree of complementation of the crc-1 mutant
phenotype in carpel fusion, carpel form and nectary development. KEY RESULTS: The
CRC orthologue from the basal angiosperm Amborella trichopoda partially
complemented the crc-1 mutant phenotype in carpels, but not in nectaries. The CRC
orthologue from rice partially complemented all aspects of the crc-1 mutant
phenotype. Though most non-CRC YABBY coding sequences did not complement crc-1
mutant phenotypes, YABBY2 (YAB2) proved to be an exception. CONCLUSIONS: The data
support a hypothesized ancestral role for CRC in carpel development and suggest
that novel roles for CRC orthologues in monocots and in core eudicots resulted
principally from molecular changes other than those affecting their coding
sequences.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17650514 [PubMed - indexed for MEDLINE]

411: Tsitol Genet. 2007 May-Jun;41(3):72-81.

[Investigation of genetically modified soybean biosafety in the center of the
origin and diversity of Russian Far East]

[Article in Russian]

Nedoluzhko AV, Dorokhov DB.

Wild soybean (Glycine soja Sieb. & Zucc.) is the nearest relative of a soybean
crop (Glycine max (L.) Merr.). Study of population genetic structure of
wild-growing relatives ofgenetically modified (GM) plants in the centers of their
origin is one of the main procedures before introduction of GM crops in these
areas. We have studied genetic variability of nine wild growing soya populations
of Primorye Territory using RAPD analysis. The level of G. soja genetic
variability was considerably higher than that of G. max. We have analyzed
phylogenetic relationships in the genus Glycine subgenus Soja using RAPD markers.
Our data confirm validity of allocation G. gracilis in a rank of a species.

Publication Types:
English Abstract

PMID: 17649627 [PubMed - indexed for MEDLINE]

412: Tsitol Genet. 2007 May-Jun;41(3):55-61.

[Experimental models of transgenic plants promising for the modern technologies]

[Article in Russian]

Abdeeva IA, Goldenkova-Pavlova IV, Mokriakova MV, Volkova LV, Bogush VG, Sidoruk
KV, Iur'eva NO, Debabov VG, Piruzian ES.

Transgenic popato plants have been created which express recombinant proteins,
analogues of spidroin 1, the protein of the cobweb skeleton thread. Expression of
the hybrid spidroin 1 genes possessing some repeated sequences retains both in
the model test-tube-growing plants and in the crops. Expression level of the
synthetic spidroin 1 genes and the level of accumulation of their products in
plants depend on the type of promoter, number of repeats, organ specificity and
plant species but not on the duration of plant material storage. The results show
that the strategy based on constuction and expression of hybrid proteins which
include the reporter protein makes it easier to select and analyse expression of
hybrid proteins in transgenic organisms.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 17649625 [PubMed - indexed for MEDLINE]

413: Tsitol Genet. 2007 May-Jun;41(3):44-9.

[Experimental models for creation of transgenic plants resistant to stress
factors]

[Article in Russian]

Goldenkova-Pavlova IV, Mirakhorli N, Maali AR, Isaenko E, Kartel' NA, Iur'eva NO,
Abdeeva IA.

Experimental models of the potato primary transgenic plants which express the
hybrid gene cry3aM-licBM2 have been created. Modecular analysis and the biotests
of the experimental models allow proposing a new system of cry genes expression
in plants. The system is based on the expression of hybrid genes possessing the
sequence of reporter lichenase gene and the use as a regulator element of a
light-induced promoter providing preferential expression of the controled genes
only in green plant tissues (leaves)--the target tissues for pests. The lichanase
presence in hybrid proteins facilitates selection and analysis of the expression
level of the hybris proteins in transgenic organisms. Basing on the lichenase
properties in hybrid proteins it seems possible to use this reporter system for
transgene monitoring in agrocoenosis as this system is rather simple and precise
and does not need large material and time expenses.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 17649623 [PubMed - indexed for MEDLINE]

414: Tsitol Genet. 2007 May-Jun;41(3):10-2.

[The global status of the commercialized biotechnological/genetically modified
crops: 2006]

[Article in Russian]

Clive J.

PMID: 17649619 [PubMed - indexed for MEDLINE]

415: Tsitol Genet. 2007 May-Jun;41(3):3-8; discussion 8-9.

[International symposium "Biosafety issues in implementation of genetically
modified organisms: new research approaches, regulation and public perception"
and its appeal for support of agricultural biology development. May 10-14, 2006.
Ialta, Ukraine]

[Article in Russian]

Blium IaB.

Publication Types:
Congresses

PMID: 17649618 [PubMed - indexed for MEDLINE]

416: Plant Biotechnol J. 2007 Sep;5(5):555-69. Epub 2007 Jul 21.

Improving containment strategies in biopharming.

Murphy DJ.

Biotechnology Unit, Division of Biological Sciences, University of Glamorgan,
Treforest, CF37 1DL, UK. dmurphy2@glam.ac.uk

This review examines the challenges of segregating biopharmed crops expressing
pharmaceutical or veterinary agents from mainstream crops, particularly those
destined for food or feed use. The strategy of using major food crops as
production vehicles for the expression of pharmaceutical or veterinary agents is
critically analysed in the light of several recent episodes of contamination of
the human food chain by non-approved crop varieties. Commercially viable
strategies to limit or avoid biopharming intrusion into the human food chain
require the more rigorous segregation of food and non-food varieties of the same
crop species via a range of either physical or biological methods. Even more
secure segregation is possible by the use of non-food crops, non-crop plants or
in vitro plant cultures as production platforms for biopharming. Such platforms
already under development range from outdoor-grown Nicotiana spp. to
glasshouse-grown Arabidopsis, lotus and moss. Amongst the more effective methods
for biocontainment are the plastid expression of transgenes, inducible and
transient expression systems, and physical containment of plants or cell
cultures. In the current atmosphere of heightened concerns over food safety and
biosecurity, the future of biopharming may be largely determined by the extent to
which the sector is able to maintain public confidence via a more considered
approach to containment and security of its plant production systems.

Publication Types:
Review

PMID: 17645439 [PubMed - indexed for MEDLINE]

417: Phytochemistry. 2007 Nov;68(21):2660-9. Epub 2007 Jul 17.

Tomato phenylacetaldehyde reductases catalyze the last step in the synthesis of
the aroma volatile 2-phenylethanol.

Tieman DM, Loucas HM, Kim JY, Clark DG, Klee HJ.

Horticultural Sciences Department, University of Florida, Gainesville, FL 32611,
United States.

The volatile compounds, 2-phenylacetaldehyde and 2-phenylethanol, are important
for the aroma and flavor of many foods, such as ripe tomato fruits, and are also
major constituents of scent of many flowers, most notably roses. While much work
has gone into elucidating the pathway for 2-phenylethanol synthesis in bacteria
and yeast, the pathways for synthesis in plants are not well characterized. We
have identified two tomato enzymes (LePAR1 and LePAR2) that catalyze the
conversion of 2-phenylacetaldehyde to 2-phenylethanol: LePAR1, a member of the
large and diverse short-chain dehydrogenase/reductase family, strongly prefers
2-phenylacetaldehyde to its shorter and longer homologues (benzaldehyde and
cinnamaldehyde, respectively) and does not catalyze the reverse reaction at a
measurable rate; LePAR2, however, has similar affinity for 2-phenylacetaldehyde,
benzaldehyde and cinnamaldehyde. To confirm the activity of these enzymes in
vivo, LePAR1 and LePAR2 cDNAs were individually expressed constitutively in
petunia. While wild type petunia flowers emit relatively high levels of
2-phenylacetaldehyde and lower levels of 2-phenylethanol, flowers from the
transgenic plants expressing LePAR1 or LePAR2 had significantly higher levels of
2-phenylethanol and lower levels of 2-phenylacetaldehyde. The in vivo alteration
of volatile emissions is an important step toward altering aroma volatiles in
plants.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17644147 [PubMed - indexed for MEDLINE]

418: Plant Cell Rep. 2007 Nov;26(11):2017-26. Epub 2007 Jul 20.

Overexpression of wheat dehydrin DHN-5 enhances tolerance to salt and osmotic
stress in Arabidopsis thaliana.

Brini F, Hanin M, Lumbreras V, Amara I, Khoudi H, Hassairi A, Pagö¨s M, Masmoudi
K.

Plant Molecular Genetics Unit, Centre of Biotechnology of Sfax, B.P'K', 3038,
Sfax, Tunisia.

Late Embryogenesis Abundant (LEA) proteins are associated with tolerance to
water-related stress. A wheat (Triticum durum) group 2 LEA proteins, known also
as dehydrin (DHN-5), has been previously shown to be induced by salt and abscisic
acid (ABA). In this report, we analyze the effect of ectopic expression of Dhn-5
cDNA in Arabidopsis thaliana plants and their response to salt and osmotic
stress. When compared to wild type plants, the Dhn-5 transgenic plants exhibited
stronger growth under high concentrations of NaCl or under water deprivation, and
showed a faster recovery from mannitol treatment. Leaf area and seed germination
rate decreased much more in wild type than in transgenic plants subjected to salt
stress. Moreover, the water potential was more negative in transgenic than in
wild type plants. In addition, the transgenic plants have higher proline contents
and lower water loss rate under water stress. Also, Na(+) and K(+) accumulate to
higher contents in the leaves of the transgenic plants. Our data strongly support
the hypothesis that Dhn-5, by its protective role, contributes to an improved
tolerance to salt and drought stress through osmotic adjustment.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17641860 [PubMed - indexed for MEDLINE]

419: Environ Biosafety Res. 2006 Oct-Dec;5(4):237-8. Epub 2007 Jul 20.

The Japanese experience with the Blue Book and subsequent activities in
environmental biosafety of GM crops.

Hayashi K.

Society for Techno-Innovation of Agriculture, Forestry and Fisheries, Sankaido
Bldg. 7F, 1-9-13 Akasaka, Minato-ku, Tokyo 107-0052, Japan. hayashi@staff.or.jp

The Blue Book made a big contribution to the development of the Japanese Ministry
of Agriculture, Forestry and Fisheries (MAFF) guidelines established in 1991 for
almost all of the basic issues. However, the MAFF guidelines could not
sufficiently cover some important areas that the Blue Book addressed well, such
as potential consequences. This gap has been recovered substantially by a new law
established in 2003. Japan still faces several important issues, including
assessment of stacked products, potential consequences, comparative assessment,
assessment of imported GM commodities and movement of concerned groups.

PMID: 17640516 [PubMed - indexed for MEDLINE]

420: Environ Biosafety Res. 2006 Oct-Dec;5(4):227-31. Epub 2007 Jul 20.

Perspective on OECD activities from a non-member country.

Alexandrova N, Atanassov A.

Agrobioinstitute, 8, Dragan Tzankov, Sofia 1164, Bulgaria.

The OECD Blue Book, "Recombinant DNA: Safety Considerations" was published in
1986. The developed principles and concepts on the stepwise and case-by-case
approach for risk assessment in the Blue Book have been used as a foundation for
building national biosafety frameworks and international instruments for the
regulation of the products of modern biotechnology. Twenty years after the Blue
Book was published, OECD continues its activities on unique identifier systems,
information-sharing, consensus documents for the biology of crops, trees and
microorganisms with respect to harmonization of regulatory oversight and those of
novel food and feed safety. These activities benefit, without any doubt, the
international community at large, including OECD non-member countries. In order
to strengthen its position in the international arena and to better respond to
the needs of the changing world, OECD would be encouraged to participate in a
more active manner in the technology transfer process and co-existence debate,
together with continuing the organization's efforts on information-sharing and
harmonization in the field of biotechnology and biosafety.

PMID: 17640514 [PubMed - indexed for MEDLINE]

421: Environ Biosafety Res. 2006 Oct-Dec;5(4):219-22. Epub 2007 Jul 20.

The impact of the OECD on the development of national/international risk/safety
assessment frameworks.

Gaugitsch H.

Environmental Impact Assessment and Biosafety, Federal Environment Agency,
Umweltbundesamt, Spittelauer Laende 5, A-1090 Vienna, Austria.
helmut.gaugitsch@umweltbundesamt.at

The role of OECD in the development of national and international risk/safety
assessment frameworks is presented and discussed. The most relevant OECD bodies
in this context have contributed a lot to the development of international
biosafety frameworks, inter alia by organizing international conferences in the
areas of food/feed and environmental safety of GMOs, focusing on practical and
current scientific issues. The OECD Consensus Documents as well as the OECD
Product Database have provided a good basis for risk/safety assessment frameworks
and their implementation. The relevance for the OECD work in the international
area is discussed, in particular, with respect to the work undertaken under the
Cartagena Protocol on Biosafety, the Codex Alimentarius Commission and its work
in biotechnology, as well as under the International Plant Protection Convention.
An outlook and suggestions for future directions of OECD work in this area are
presented.

PMID: 17640512 [PubMed - indexed for MEDLINE]

422: Environ Biosafety Res. 2006 Oct-Dec;5(4):213-8. Epub 2007 Jul 20.

Basic framework for risk assessment for transgenic plants developed by the OECD:
20 years after the OECD "Blue Book".

Bergmans H.

National Institute for Public Health and the Environment, GMO Office, PO Box 1,
Bilthoven 3720BA, The Netherlands. hans.bergmans@rivm.nl

Publication Types:
Review

PMID: 17640511 [PubMed - indexed for MEDLINE]

423: Environ Biosafety Res. 2006 Oct-Dec;5(4):207-9. Epub 2007 Jul 20.

Beyond the Blue Book. Framework for Risk/Safety Assessment of Transgenic Plants.
An overview of the workshop.

Kobayashi M, Kearns P.

OECD Biosafety Team--Paris, 2 rue Andrö© Pascal, 75775 Paris, France.
masatoshi.kobayashi@oecd.org

Publication Types:
Congresses

PMID: 17640509 [PubMed - indexed for MEDLINE]

424: Environ Biosafety Res. 2006 Oct-Dec;5(4):201-3. Epub 2007 Jul 20.

9th International Symposium on the Biosafety of Genetically Modified Organisms.
Session VII: Risk management and monitoring.

Schiemann J.

Institute for Plant Virology, Microbiology and Biosafety, Federal Biological
Research Centre for Agriculture and Forestry (BBA), Messeweg 11-12, 38104
Braunschweig, Germany. j.schiemann@bba.de

Biosafety regulatory frameworks are intended to serve as mechanisms for ensuring
the safe use of biotechnology products without imposing unacceptable risk to
human health or the environment, or unintended constraints to technology
transfer. In several regulatory systems GMO risk assessment has been separated
from GMO risk management. As a consequence, risk assessment can be performed on a
purely scientific basis, whereas risk management can take additional aspects
(e.g. socio-economic or ethical) into consideration. For instance, the European
Food Safety Authority (EFSA), the keystone of European Union risk assessment
regarding food and feed safety, provides independent scientific advice and clear
communication on existing and emerging risks in close collaboration with national
authorities and in open consultation with its stakeholders. Risk management
measures are not within the remit of EFSA, and remain the responsibility of the
European Commission and Member States.

Publication Types:
Congresses

PMID: 17640508 [PubMed - indexed for MEDLINE]

425: Environ Biosafety Res. 2006 Oct-Dec;5(4):197-9. Epub 2007 Jul 20.

9th International Symposium on the Biosafety of Genetically Modified Organisms.
Session VI: Estimating likelihood and exposure, Part II.

Quemada H.

Department of Biological Sciences, Wood Hall, Western Michigan University,
Kalamazoo, Michigan 49004, USA. hector.quemada@wmich.edu

Publication Types:
Congresses

PMID: 17640507 [PubMed - indexed for MEDLINE]

426: Environ Biosafety Res. 2006 Oct-Dec;5(4):193-5. Epub 2007 Jul 20.

9th International Symposium on the Biosafety of Genetically Modified Organisms.
Session V: Estimating likelihood and exposure.

Lentini Z.

CIAT (International Center for Tropical Agriculture), km 17 Recta Cali-Palmira,
Cali, A.A. 6713, Colombia. z.lentini@cgiar.org

Publication Types:
Congresses

PMID: 17640506 [PubMed - indexed for MEDLINE]

427: Environ Biosafety Res. 2006 Oct-Dec;5(4):189-92. Epub 2007 Jul 20.

9th International Symposium on the Biosafety of Genetically Modified Organisms.
Session IV: Identifying and defining hazards and potential consequences III:
Concepts for problem formulation and non-target risk assessment.

Alvarez-Morales A.

Department of Genetic Engineering, Center for Research and Advanced Studies
(Cinvestav), Irapuato, KM 9.6 Libramiento Norte, Irapuato 36821, Mexico.
aalvarez@ira.cinvestav.mx

Publication Types:
Congresses

PMID: 17640505 [PubMed - indexed for MEDLINE]

428: Environ Biosafety Res. 2006 Oct-Dec;5(4):187-8. Epub 2007 Jul 20.

9th International Symposium on the Biosafety of Genetically Modified Organisms.
Session III: Identifying and defining hazards and potential consequences II.

Sweet J.

Sweet Environmental Consultants, 6 The Green, Willingham, Cambridge CB4 5JA, UK.
jeremysweet303@aol.com

Publication Types:
Congresses

PMID: 17640504 [PubMed - indexed for MEDLINE]

429: Environ Biosafety Res. 2006 Oct-Dec;5(4):183-6. Epub 2007 Jul 20.

9th International Symposium on the Biosafety of Genetically Modified Organisms.
Session II: Identifying and defining hazards and potential consequences I:
Concepts for problem formulation and non-target risk assessment.

Bigler F.

Agroscope Reckenholz-Taeknikon ART, Swiss Agricultural Research,
Reckenholzstrasse 191, 8046 Zurich, Switzerland. franz.bigler@art.admin.ch

The scientific organizers of the symposium put much emphasis on the
identification and definition of hazard and the potential consequences thereof
and three full sessions with a total of 13 presentations encompassing a wide
range of related themes were planned for this topic. Unfortunately, one talk had
to be cancelled because of illness of the speaker (BM Khadi, India). Some
presentations covered conceptual approaches for environmental risk assessment
(ERA) of GM plants (problem formulation in the risk assessment framework,
familiarity approach, tiered and methodological frameworks, non-target risk
assessment) and the use of models in assessing invasiveness and weediness of GM
plants. Other presentations highlighted the lessons learned for future ERA from
case studies and commercialized GM crops, and from monitoring of unintended
releases to the environment. When the moderators of the three sessions came
together after the presentations to align their summaries, there was an obvious
need to restructure the 12 presentations in a way that allowed for a consistent
summarizing discussion. The following new organization of the 12 talks was
chosen: (1) Concepts for problem formulation and non-target risk assessment, (2)
Modeling as a tool for predicting invasiveness of GM plants, (3) Case-studies of
ERA of large-scale release, (4) Lessons learned for ERA from a commercialized GM
plant, (5) Monitoring of unintended release of Bt maize in Mexico. The new
thematic structure facilitates a more in-depth discussion of the presentations
related to a specific topic, and the conclusions to be drawn are thus more
consistent. Each moderator agreed to take responsibility for summarizing one or
more themes and to prepare the respective report.

Publication Types:
Congresses

PMID: 17640503 [PubMed - indexed for MEDLINE]

430: Mol Nutr Food Res. 2007 Aug;51(8):946-55.

Serum testing of genetically modified soybeans with special emphasis on potential
allergenicity of the heterologous protein CP4 EPSPS.

Hoff M, Son DY, Gubesch M, Ahn K, Lee SI, Vieths S, Goodman RE, Ballmer-Weber BK,
Bannon GA.

Department of Allergology, Paul Ehrlich Institute, Langen, Germany.

Roundup Ready soy contains the CP4-enolpyruvylshikimate-3-phosphate synthase (CP4
EPSPS) protein. Serum IgE from two distinct populations of soy-allergic patients
were recruited to determine their IgE-binding specificity. One population
consisted of 10 adult patients from Europe, whose primary diagnosis was soy food
allergy with some also having mite allergy. In addition, 6 primarily
mite-allergic, 6 food-allergic (celery, carrot, milk, shrimp, walnut, and apple),
and 5 non-allergic patients were tested. Another population consisted of 13
children from Korea, whose primary diagnosis was atopic dermatitis and
secondarily soy and egg sensitization. In addition, 11 non-allergic patients were
tested. Each patient population was extensively characterized with respect to
clinical symptoms, specific IgE (CAP) scores, and total IgE. Immunoblots and
ELISA assays were developed using serum IgE from these patients and soy extracts,
CP4 EPSPS, rice extract, ovalbumin, rubisco, purified major peanut allergen Ara h
2, the putative soy allergen Gly m Bd 30k and mite allergen Der f 2 proteins as
the intended targets. Immunoblot results indicated that soy-allergic patients
bound soy extracts but did not specifically bind rubisco or CP4 EPSPS. ELISA
results were in general agreement with the immunoblot results except that rubisco
bound significant quantities of serum IgE from some patients. These results
indicate that the CP4 EPSPS protein does not bind significant quantities of IgE
from two geographically distinct sensitive populations and there is no evidence
for an increased allergenic potential of this biotech protein.

PMID: 17639514 [PubMed - indexed for MEDLINE]

431: Plant Cell Rep. 2007 Nov;26(11):1999-2008. Epub 2007 Jul 18.

LeERF1 positively modulated ethylene triple response on etiolated seedling, plant
development and fruit ripening and softening in tomato.

Li Y, Zhu B, Xu W, Zhu H, Chen A, Xie Y, Shao Y, Luo Y.

Laboratory of Fruit Physiology and Molecular Biology, College of Food Science and
Nutritional Engineering, China Agricultural University, Beijing 100083, People's
Republic of China.

To study the function of LeERF1 in ethylene triple response on etiolated
seedling, plant development and fruit ripening and softening, LeERF1 gene was
introduced into tomato (Lycopersicon esculentum cv. No. 4 Zhongshu) through
Agrobacterium-mediated transformation. The sense LeERF1 and anti-sense LeERF1
transgenic tomato were obtained. Overexpression of LeERF1 in tomato caused the
typical ethylene triple response on etiolated seedling. In the adult stage,
35S::LeERF1 resulted in morphological changes in the leaves of the LeERF1-sn
lines. Anti-sense LeERF1 fruits had longer shelf life compared with wild-type
tomato. The results of this manuscript indicated that LeERF1 positively mediated
the ethylene signals, while the function of LeERF1 was verified for the first
time to be positively related with ethylene triple response on etiolated
seedling, plant development and fruit ripening and softening using LeERF1-sn, wt
and LeERF1-as tomato.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17639404 [PubMed - indexed for MEDLINE]

432: Transgenic Res. 2008 Jun;17(3):393-402. Epub 2007 Jul 19.

Impact of genetic structures on haploid genome-based quantification of
genetically modified DNA: theoretical considerations, experimental data in MON
810 maize kernels (Zea mays L.) and some practical applications.

Zhang D, Corlet A, Fouilloux S.

GEVES Domaine du Magneraud, Laboratoire BioGEVES, B.P. 52, Surgeres, 17700,
France, david.zhang@geves.fr.

Real-time Polymerase Chain Reaction (PCR) based assays are widely used to
estimate the content of genetically modified (GM) materials in food, feed and
seed. It has been known that the genetic structures of the analyte can
significantly influence the GM content expressed by the haploid genome (HG) %
estimated using real-time PCR assays; this kind of influence is also understood
as the impact of biological factors. The influence was first simulated at
theoretical level using maize as a model. We then experimentally assessed the
impact of biological factors on quantitative results, analysing by quantitative
real-time PCR six maize MON 810 hybrid kernels with different genetic structures:
(1) hemizygous from transgenic male parent, (2) hemizygous from transgenic female
parent and (3) homozygous at the transgenic locus. The results obtained in the
present study showed clear influences of biological factors on GM DNA
quantification: 1% of GM materials by weight (wt) for the three genetic
structures contained 0.39, 0.55 and 1.0% of GM DNA by HG respectively, from
quantitative real-time PCR analyses. The relationships between GM wt% and GM HG%
can be empirically established as: (1) in the case of the presence of a single GM
trait: GM HG% = GM wt% x (0.5 +/- 0.167Y), where Y is the endosperm DNA content
(%) in the total DNA of a maize kernel, (2) in the case of the presence of
multiple GM traits: GM HG% = N x GM wt% x (0.5 +/- 0.167Y), where N is the number
of GM traits (stacked or not) present in an unknown sample. This finding can be
used by stakeholders related to GMO for empirical prediction from one unit of
expression to another in the monitoring of seed and grain production chains.
Practical equations have also been suggested for haploid copy number
calculations, using hemizygous GM materials for calibration curves.

PMID: 17638110 [PubMed - in process]

433: Plant J. 2007 Sep;51(6):978-90. Epub 2007 Jul 17.

Genetic and molecular requirements for function of the Pto/Prf effector
recognition complex in tomato and Nicotiana benthamiana.

Balmuth A, Rathjen JP.

The Sainsbury Laboratory, Norwich Research Park, Colney Lane, Norwich NR4 7UH,
UK.

The Pto gene of tomato (Solanum lycopersicum) confers specific recognition of the
unrelated bacterial effector proteins AvrPto and AvrPtoB. Pto resides in a
constitutive molecular complex with the nucleotide binding site-leucine rich
repeats protein Prf. Prf is absolutely required for specific recognition of both
effectors. Here, using stable transgenic lines, we show that expression of Pto
from its genomic promoter in susceptible tomatoes was sufficient to complement
recognition of Pseudomonas syringae pv. tomato (Pst) bacteria expressing either
avrPto or avrPtoB. Pto kinase activity was absolutely required for specific
immunity. Expression of the Pto N-myristoylation mutant, pto(G2A), conferred
recognition of Pst (avrPtoB), but not Pst (avrPto), although bacterial growth in
these lines was intermediate between resistant and susceptible lines.
Overexpression of pto(G2A) complemented recognition of avrPto. Transgenic tomato
plants overexpressing wild-type Pto exhibited constitutive growth phenotypes, but
these were absent in lines overexpressing pto(G2A). Therefore, Pto myristoylation
is a quantitative factor for effector recognition in tomato, but is absolutely
required for overexpression phenotypes. Native expression of Pto in the
heterologous species Nicotiana benthamiana did not confer resistance to P.
syringae pv. tabaci (Pta) expressing avrPto or avrPtoB, but recognition of both
effectors was complemented by Prf co-expression. Thus, specific resistance
conferred solely by Pto in N. benthamiana is an artefact of overexpression.
Finally, pto(G2A) did not confer recognition of either avrPto or avrPtoB in N.
benthamiana, regardless of the presence of Prf. Thus, co-expression of Prf in N.
benthamiana complements many but not all aspects of normal Pto function.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17635766 [PubMed - indexed for MEDLINE]

434: J Neurosci. 2007 Jul 18;27(29):7640-7.

D1 dopamine receptor dDA1 is required in the mushroom body neurons for aversive
and appetitive learning in Drosophila.

Kim YC, Lee HG, Han KA.

Department of Biology and The Huck Institute Neuroscience Graduate Program,
Pennsylvania State University, University Park, Pennsylvania 16802, USA.

Drosophila has robust behavioral plasticity to avoid or prefer the odor that
predicts punishment or food reward, respectively. Both types of plasticity are
mediated by the mushroom body (MB) neurons in the brain, in which various
signaling molecules play crucial roles. However, important yet unresolved
molecules are the receptors that initiate aversive or appetitive learning
cascades in the MB. We have shown previously that D1 dopamine receptor dDA1 is
highly enriched in the MB neuropil. Here, we demonstrate that dDA1 is a key
receptor that mediates both aversive and appetitive learning in pavlovian
olfactory conditioning. We identified two mutants, dumb1 and dumb2, with abnormal
dDA1 expression. When trained with the same conditioned stimuli, both dumb
alleles showed negligible learning in electric shock-mediated conditioning while
they exhibited moderately impaired learning in sugar-mediated conditioning. These
phenotypes were not attributable to anomalous sensory modalities of dumb mutants
because their olfactory acuity, shock reactivity, and sugar preference were
comparable to those of control lines. Remarkably, the dumb mutant's impaired
performance in both paradigms was fully rescued by reinstating dDA1 expression in
the same subset of MB neurons, indicating the critical roles of the MB dDA1 in
aversive as well as appetitive learning. Previous studies using dopamine receptor
antagonists implicate the involvement of D1/D5 receptors in various pavlovian
conditioning tasks in mammals; however, these have not been supported by the
studies of D1- or D5-deficient animals. The findings described here unambiguously
clarify the critical roles of D1 dopamine receptor in aversive and appetitive
pavlovian conditioning.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17634358 [PubMed - indexed for MEDLINE]

435: Plant Cell Physiol. 2007 Aug;48(8):1081-91. Epub 2007 Jul 18.

An aluminum-activated citrate transporter in barley.

Furukawa J, Yamaji N, Wang H, Mitani N, Murata Y, Sato K, Katsuhara M, Takeda K,
Ma JF.

Research Institute for Bioresources, Okayama University, Chuo, Kurashiki,
Okayama, 710-0046, Japan.

Soluble ionic aluminum (Al) inhibits root growth and reduces crop production on
acid soils. Al-resistant cultivars of barley (Hordeum vulgare L.) detoxify Al by
secreting citrate from the roots, but the responsible gene has not been
identified yet. Here, we identified a gene (HvAACT1) responsible for the
Al-activated citrate secretion by fine mapping combined with microarray analysis,
using an Al-resistant cultivar, Murasakimochi, and an Al-sensitive cultivar,
Morex. This gene belongs to the multidrug and toxic compound extrusion (MATE)
family and was constitutively expressed mainly in the roots of the Al-resistant
barley cultivar. Heterologous expression of HvAACT1 in Xenopus oocytes showed
efflux activity for (14)C-labeled citrate, but not for malate. Two-electrode
voltage clamp analysis also showed transport activity of citrate in the
HvAACT1-expressing oocytes in the presence of Al. Overexpression of this gene in
tobacco enhanced citrate secretion and Al resistance compared with the wild-type
plants. Transiently expressed green fluorescent protein-tagged HvAACT1 was
localized at the plasma membrane of the onion epidermal cells, and immunostaining
showed that HvAACT1 was localized in the epidermal cells of the barley root tips.
A good correlation was found between the expression of HvAACT1 and citrate
secretion in 10 barley cultivars differing in Al resistance. Taken together, our
results demonstrate that HvAACT1 is an Al-activated citrate transporter
responsible for Al resistance in barley.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17634181 [PubMed - indexed for MEDLINE]

436: Plant Cell Physiol. 2007 Aug;48(8):1219-28. Epub 2007 Jul 18.

A wilty mutant of rice has impaired hydraulic conductance.

Koizumi K, Ookawa T, Satoh H, Hirasawa T.

Graduate School of Agriculture, Tokyo University of Agriculture and Technology,
Fuchu, Tokyo, 183-8509, Japan.

The rice CM2088 mutant is the wilty phenotype and wilts markedly under
well-watered sunny conditions. The leaf water potential and epidermal (mainly
stomatal) conductance of CM2088 plants decreased significantly under conditions
that induced intense transpiration, as compared with those of wild-type plants,
revealing that the wilty phenotype was not the result of abnormal stomatal
behavior but was due to an increase in resistance to water transport. The
resistance to water transport was dramatically elevated in the node and the
sheath and blade of a leaf of the mutant, but not in the root or stem. The
diameter of xylem vessels in the large vascular bundles of the leaf sheath and
the internode tended to be small, and the numbers of vessel elements with
narrowed or scalariform perforation plates in the leaf blade and sheath were
greater in the mutant than in the wild type. Most xylem vessels were occluded,
with air bubbles in the leaf sheath of the mutant during the midday hours under
intense transpiration conditions, while no bubbles were observed in plants that
were barely transpiring, revealing that the significant increase in resistance to
water transport was a result of the cavitation. The additive effects of
cavitation in xylem vessels and the decreased diameter and deformed plates of
vessel elements might be responsible for the wilty phenotype of CM2088.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17634180 [PubMed - indexed for MEDLINE]

437: Plant Cell Rep. 2007 Nov;26(11):1967-75. Epub 2007 Jul 14.

Soybean dwarf virus-resistant transgenic soybeans with the sense coat protein
gene.

Tougou M, Yamagishi N, Furutani N, Shizukawa Y, Takahata Y, Hidaka S.

National Agricultural Research Center for Tohoku Region, Morioka, Iwate 020-0198,
Japan. tougou@affrc.go.jp

We transformed a construct containing the sense coat protein (CP) gene of Soybean
dwarf virus (SbDV) into soybean somatic embryos via microprojectile bombardment
to acquire SbDV-resistant soybean plants. Six independent T(0) plants were
obtained. One of these transgenic lines was subjected to further extensive
analysis. Three different insertion patterns of Southern blot hybridization
analysis in T(1) plants suggested that these insertions introduced in T(0) plants
were segregated from each other or co-inherited in T(1) progenies. These
insertions were classified into two types, which overexpressed SbDV-CP mRNA and
accumulated SbDV-CP-specific short interfering RNA (siRNA), or repressed
accumulation of SbDV-CP mRNA and siRNA by RNA analysis prior to SbDV inoculation.
After inoculation of SbDV by the aphids, most T(2) plants of this transgenic line
remained symptomless, contained little SbDV-specific RNA by RNA dot-blot
hybridization analysis and exhibited SbDV-CP-specific siRNA. We discuss here the
possible mechanisms of the achieved resistance, including the RNA silencing.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17632723 [PubMed - indexed for MEDLINE]

438: Biochem Biophys Res Commun. 2007 Sep 7;360(4):880-4. Epub 2007 Jul 9.

Identification of multiple highly similar XIP-type xylanase inhibitor genes in
hexaploid wheat.

Takahashi-Ando N, Inaba M, Ohsato S, Igawa T, Usami R, Kimura M.

Plant & Microbial Metabolic Engineering Research Unit and Laboratory for
Remediation Research, Discovery Research Institute (DRI) and Plant Science Center
(PSC1), RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.

In hexaploid wheat, Xip-I is the only XIP-type xylanase inhibitor gene whose
expression and function have been characterized in detail. Here we demonstrate
the existence of new XIP-type genes with the identification of Xip-R1 and Xip-R2
in the root cDNAs. Southern blot analysis with the Xip-R1 probe revealed that
XIP-type genes comprised a significantly greater gene family than previously
speculated on in studies with the Xip-I probe. The transcript level of Xip-R
genes was increased upon an inoculation with Erysiphe graminis in the leaves, but
not with Fusarium graminearum in the spikelets. RT-PCR with the RNA samples
followed by extensive sequencing of the cloned amplified products revealed the
presence of 12 highly similar Xip-R genes. Among these genes, Xip-R1 was the only
predominant Xip-R family member induced to express in response to E. graminis.
XIP-R1 was located in the apoplastic space and inhibited family 11 xylanases, but
the protein did not show chitinolytic activity. These results suggest that
hexaploid wheat has a large family of XIPs in its genome, but that only some of
them are expressed for plant defense in limited tissues.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17631271 [PubMed - indexed for MEDLINE]

439: J Exp Bot. 2007;58(11):2909-15. Epub 2007 Jul 13.

Overexpression of the OsZIP4 zinc transporter confers disarrangement of zinc
distribution in rice plants.

Ishimaru Y, Masuda H, Suzuki M, Bashir K, Takahashi M, Nakanishi H, Mori S,
Nishizawa NK.

Department of Global Agricultural Sciences, Graduate School of Agricultural and
Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657
Japan.

Zinc (Zn), an essential nutrient in cells, plays a vital role in controlling
cellular processes such as growth, development, and differentiation. Although the
mechanisms of Zn translocation in rice plants (Oryza sativa) are not fully
understood, it has recently received increased interest. OsZIP4 is a Zn
transporter that localizes to apical cells. Transgenic rice plants overexpressing
the OsZIP4 gene under the control of the cauliflower mosaic virus (CaMV) 35S
promoter were produced. The Zn concentration in roots of 35S-OsZIP4 transgenic
plants was 10 times higher than in those of vector controls, but it was five
times lower in shoots. The Zn concentration in seeds of 35S-OsZIP4 plants was
four times lower compared with vector controls. Northern blot analysis and
quantitative real-time reverse transcription-PCR revealed transcripts of OsZIP4
expression driven by the CaMV 35S promoter in roots and shoots of 35S-OsZIP4
plants, but levels of endogenous OsZIP4 transcripts were low in roots and high in
shoots compared with vector controls. Microarray analysis revealed that the genes
expressed in shoots of 35S-OsZIP4 plants coincided with those induced in shoots
of Zn-deficient plants. These results indicate that constitutive expression of
OsZIP4 changes the Zn distribution within rice plants, and that OsZIP4 is a
critical Zn transporter that must be strictly regulated.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17630290 [PubMed - indexed for MEDLINE]

440: Vaccine. 2007 Aug 21;25(34):6373-80. Epub 2007 Jun 27.

Recombinant rotavirus inner core proteins produced in the milk of transgenic
rabbits confer a high level of protection after intrarectal delivery.

Soler E, Parez N, Passet B, Dubuquoy C, Riffault S, Pillot M, Houdebine LM,
Schwartz-Cornil I.

Biologie du Dö©veloppement et de la Reproduction, Domaine de Vilvert, 78352
Jouy-en-Josas Cedex, France.

Development of a safe, cheap and efficient vaccine against rotavirus is important
to reduce the morbidity and mortality associated with gastroenteritis in infants
worldwide. High quantities of two inner core rotavirus-derived proteins (VP2 and
a nonglycosylated mutant VP6 (VP6(NG)) from the RF81 bovine strain) were produced
in the milk of transgenic rabbits. We show here that rectal administration of
partially purified milk-derived VP2 and VP6(NG) proteins with the detoxified
LT(R192G) adjuvant almost completely prevented fecal shedding induced by a highly
infectious challenge in mice with the murine ECw strain. The vaccine generated
rotavirus-specific fecal secretory IgA, systemic IgG and IgA and a
rotavirus-specific Th1 response. We thus demonstrate in clinically feasible
settings that mass production of viral protein in transgenic milk is a promising
way to generate subunit vaccine against rotavirus.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17629366 [PubMed - indexed for MEDLINE]

441: Planta. 2007 Nov;226(6):1353-62. Epub 2007 Jul 13.

EgAP2-1, an AINTEGUMENTA-like (AIL) gene expressed in meristematic and
proliferating tissues of embryos in oil palm.

Morcillo F, Gallard A, Pillot M, Jouannic S, Aberlenc-Bertossi F, Collin M,
Verdeil JL, Tregear JW.

CIRAD/IRD Palm Group, UMR DAP/LIR IRD 192, Centre IRD Montpellier, BP 64501, 911
avenue Agropolis, 34394, Montpellier Cedex 5, France. morcillo@mpl.ird.fr

In order to better understand the developmental processes that govern the
formation of somatic embryos in oil palm (Elaeis guineensis Jacq.), we
investigated the transcription factor genes expressed during embryogenesis in
this species. The AP2/EREBP transcription factor family includes the AP2
subgroup, which contains several proteins that play important roles in plant
development. We identified and characterized EgAP2-1, which codes for a protein
that contains two AP2 domains similar to those of the transcription factor
BABYBOOM (BBM) and more generally AINTEGUMENTA-like (AIL) proteins of the AP2
subgroup. In a similar way to related genes from eudicots, ectopic expression of
EgAP2-1 in transgenic Arabidopsis plants alters leaf morphology and enhances
regeneration capacity. In oil palm, EgAP2-1 transcripts accumulate to the
greatest extent in zygotic embryos. This expression pattern was investigated in
more detail by in-situ hybridization, revealing that in both zygotic and somatic
embryos, EgAP2-1 expression is concentrated in proliferating tissues associated
with the early development of leaf primordia, root initials and provascular
tissues.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17628826 [PubMed - indexed for MEDLINE]

442: Ambio. 2007 Jun;36(4):359-61.

Transgenic maize containing the Cry1Ab protein ephemerally enhances soil
microbial communities.

Mulder C, Wouterse M, Rutgers M, Posthuma L.

National Institute for Public Health and the Environment, Bilthoven, The
Netherlands. Christian.Mulder@rivm.nl

PMID: 17626475 [PubMed - indexed for MEDLINE]

443: Transgenic Res. 2007 Aug;16(4):467-78. Epub 2006 Nov 25.

High level accumulation of alpha-glucan in maize kernels by expressing the gtfD
gene from Streptococcus mutans.

Zhang S, Dong JG, Wang T, Guo S, Glassman K, Ranch J, Nichols SE.

Pioneer Hi-Bred International, Inc., a DuPont company, Johnston, IA 50131, USA.
shirong.zhang@pioneer.com

Glucosyltransferases (GTFs, EC.2.4.1.5) are bacterial enzymes that catalyze the
polymerization of glucose residues from sucrose, leading to the production of
high molecular weight glucan with alpha-1,3 /alpha-1,6 linkages. Such glucans,
with many potential food and industrial applications, do not normally exist in
higher plants. We fused a mutant form of the gtfD gene from Sreptococcus mutans
with the maize (Zea mays L.) chloroplastic Brittle 1 transit peptide for
amyloplast targeting. This construct, driven by the ubiquitin promoter, was
introduced into maize by Agrobacterium-mediated transformation. We developed a
novel HPLC-based method that enabled us differentially to distinguish transgene
glucan from other endogenous polysaccharides in maize kernels. Using this method,
we screened over 100 transgenic plants for the presence of GTF-produced glucan
whose content varied between 0.8 and 14% of dry weight in the mature transgenic
seeds. The mature transgenic plants were indistinguishable from wildtype plants
in growth rate and morphology. Furthermore, starch granule size in the transgenic
maize kernel was unaffected by the accumulation of the foreign polysaccharide.
Mutation in Sh2, which encodes a subunit of ADP-glucose pyrophosphorylase, had no
effect on glucan accumulation caused by gtfD expression. Our results indicated
that high levels of novel carbohydrate polymer can be accumulated in crop plants
through transgene technology.

PMID: 17624807 [PubMed - indexed for MEDLINE]

444: Planta. 2007 Dec;227(1):1-12. Epub 2007 Jul 13.

A Ds-insertion mutant of OSH6 (Oryza sativa Homeobox 6) exhibits outgrowth of
vestigial leaf-like structures, bracts, in rice.

Park SH, Kim CM, Je BI, Park SH, Park SJ, Piao HL, Xuan YH, Choe MS, Satoh K,
Kikuchi S, Lee KH, Cha YS, Ahn BO, Ji HS, Yun DW, Lee MC, Suh SC, Eun MY, Han CD.

Division of Applied Life Science, Plant Molecular Biology and Biotechnology
Research Center, Environmental Biotechnology National Core Research Center,
Gyeongsang National University, Jinju, South Korea.

OSH6 (Oryza sativa Homeobox6) is an ortholog of lg3 (Liguleless3) in maize. We
generated a novel allele, termed OSH6-Ds, by inserting a defective Ds element
into the third exon of OSH6, which resulted in a truncated OSH6 mRNA. The
truncated mRNA was expressed ectopically in leaf tissues and encoded the
N-terminal region of OSH6, which includes the KNOX1 and partial KNOX2 subdomains.
This recessive mutant showed outgrowth of bracts or produced leaves at the basal
node of the panicle. These phenotypes distinguished it from the OSH6 transgene
whose ectopic expression led to a "blade to sheath transformation" phenotype at
the midrib region of leaves, similar to that seen in dominant Lg3 mutants.
Expression of a similar truncated OSH6 cDNA from the 35S promoter
(35S::DeltaOSH6) confirmed that the ectopic expression of this product was
responsible for the aberrant bract development. These data suggest that OSH6-Ds
interferes with a developmental mechanism involved in bract differentiation,
especially at the basal nodes of panicles.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17624547 [PubMed - indexed for MEDLINE]

445: Amino Acids. 2008 Feb;34(2):213-22. Epub 2007 Jul 12.

Regulation of aspartate-derived amino acid homeostasis in potato plants (Solanum
tuberosum L.) by expression of E. coli homoserine kinase.

Rinder J, Casazza AP, Hoefgen R, Hesse H.

Department of Molecular Physiology, Max-Planck-Institut fö¼r Molekulare
Pflanzenphysiologie, Golm, Germany.

The availability of the carbon backbone O-phosphohomoserine (OPHS) is critical to
methionine (met) and threonine (thr) synthesis. OPHS derives from homoserine and
is formed by homoserine kinase (HSK). To clarify the function of HSK in cellular
metabolism, the E. coli HSK ortholog thrB was expressed in potato plants
targeting the EcHSK protein to chloroplasts and to the cytosol. Both approaches
resulted in up to 11 times increased total HSK enzyme activity. Transgenic plants
exhibited reduced homoserine levels while met and thr did not accumulate
significantly. However, the precursor cysteine and upstream intermediates of met
such as cystathionine and homocysteine did indicating an accelerated carbon flow
towards the end products. Coincidently, plants with elevated cytosolic levels of
EcHSK exhibited a reduction in transcript levels of the endogenous HSK, as well
as of threonine synthase (TS), cystathionine beta-lyase (CbL), and met synthase
(MS). In all plants, cystathionine gamma-synthase (CgS) expression remained
relatively unchanged from wild type levels, while S-adenosylmethionine synthetase
(SAMS) expression increased. Feeding studies with externally supplied homoserine
fostered the synthesis of met and thr but the regulation of synthesis of both
amino acids retained the wild type regulation pattern. The results indicate that
excess of plastidial localised HSK activity does not influence the de novo
synthesis of met and thr. However, expression of HSK in the cytosol resulted in
the down-regulation of gene expression of pathway genes probably mediated via
OPHS. We integrated these data in a novel working model describing the regulatory
mechanism of met and thr homeostasis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17624493 [PubMed - indexed for MEDLINE]

446: Plant Cell Physiol. 2007 Aug;48(8):1192-206. Epub 2007 Jul 10.

Overexpression of TaVRN1 in Arabidopsis promotes early flowering and alters
development.

Adam H, Ouellet F, Kane NA, Agharbaoui Z, Major G, Tominaga Y, Sarhan F.

Universitö© du Quö©bec ö  Montrö©al, Dö©partement des Sciences biologiques, Case
Postale 8888, Succursale Centre-ville, Montrö©al, Quö©bec, Canada H3C 3P8.

TaVRN1, a member of the APETALA1 (AP1) subfamily of MADS-box transcription
factors, is a key gene that controls transition from vegetative to reproductive
phase in wheat. The accumulation of TaVRN1 transcripts in winter wheat probably
requires the down-regulation of TaVRT2, a MADS-box factor that binds and
represses the TaVRN1 promoter, and of the flowering repressor TaVRN2. However,
the molecular mechanisms by which TaVRN1 functions as an activator of phase
transition is unknown. To address this, a combination of gene expression and
functional studies was used. RNA in situ hybridization studies showed that TaVRN1
transcripts accumulate in all meristems and primordia associated with flower
development. An interaction screen in yeast revealed that TaVRN1 interacts with
several proteins involved in different processes of plant development such as
transcription factors, kinases and a cyclophilin. Arabidopsis plants
overexpressing TaVRN1 flower early and show various levels of modified plant
architecture. The ectopic expression causes an overexpression of the AP1 and MAX4
genes, which are associated with flowering and auxin regulation, respectively.
The induction of gene expression probably results from the binding of TaVRN1 to
CArG motifs present on the AP1 and MAX4 promoters. In contrast, Arabidopsis
plants that overexpress TaVRT2, which encodes a putative flowering repressor,
show an opposite late flowering phenotype. Together, the data provide molecular
evidence that TaVRN1 may have pleiotropic effects in various processes such as
control of axillary bud growth, transition to flowering and development of floral
organs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17623742 [PubMed - indexed for MEDLINE]

447: Theor Appl Genet. 2007 Aug;115(4):549-60. Epub 2007 Jul 11.

Investigation of rice transgene flow in compass sectors by using male sterile
line as a pollen detector.

Yuan QH, Shi L, Wang F, Cao B, Qian Q, Lei XM, Liao YL, Liu WG, Cheng L, Jia SR.

College of Life Science and Agriculture, MOE Key Lab of Tropic Biological
Resources, Hainan University, Haikou, 570228, China.

Rice is the most important staple food in the world. The rapid development of
transgenic rice and its future commercialization have raised concerns regarding
transgene flow and its potential environmental risk. It is known that rice is a
self-pollinated crop; the outcrossing rate between common cultivars is generally
less than 1%. In order to improve the detection sensitivity of rice transgene
flow, a male sterile (ms) line BoA with a high outcrossing rate was used as a
pollen detector in this study. A concentric circle design was adopted, in which
the transgenic rice B2 containing bar gene as a pollen donor was planted in the
center circle and the recipient BoA was planted in eight compass sectors. The
frequency of transgene flow in compass sectors was analyzed by continuous
sampling to generate cumulative data. The results of two years with sound
reproducibility demonstrated that the rice gene flow was closely associated with
the wind direction. According to the mean frequency of transgene flow, the eight
sectors can be divided into two groups: a higher frequency group downstream of
the prevailing wind (DPW) with a mean frequency ranging from 6.47 to 26.24%, and
a lower frequency group lateral to or upstream of the prevailing wind (UPW) with
a mean frequency of 0.39 to 3.03%. On the basis of the cumulative data, 90-96% of
the cumulative gene flow events occurred in the four DPW sectors, while it was
4-10% in the four UPW sectors. By using these systematic data, simulation models
and isograms of transgene flow in the eight compass sectors were calculated and
drawn, respectively.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17622509 [PubMed - indexed for MEDLINE]

448: J Hered. 2007 Jul-Aug;98(4):311-6. Epub 2007 Jul 9.

Introgression mapping of genes for winter hardiness and frost tolerance
transferred from Festuca arundinacea into Lolium multiflorum.

Kosmala A, Zwierzykowski Z, Zwierzykowska E, Luczak M, Rapacz M, Gasior D,
Humphreys M.

Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyå„ska 34, 60-479
Poznaå„, Poland. akos@igr.poznan.pl

Genes for winter hardiness and frost tolerance were introgressed from Festuca
arundinacea into winter-sensitive Lolium multiflorum. Two partly fertile,
pentaploid (2n = 5x = 35) F(1) hybrids F. arundinacea (2n = 6x = 42) x L.
multiflorum (2n = 4x = 28) were generated and backcrossed twice onto L.
multiflorum (2x). The backcross 1 (BC(1)) and backcross 2 (BC(2)) plants were
preselected for high vigor and good fertility, and subsequently, a total of 83
BC(2) plants were selected for winter hardiness after 2 Polish winters and by
simulated freezing tests. Genomic in situ hybridization (GISH) was performed on 6
winter-hardy plants selected after the first winter and shown to be significantly
(P < 0.05) more frost tolerant than the L. multiflorum control. Among the
analyzed BC(2) winter survivors, only diploid (2n = 2x = 14) plants were found.
Five plants carried 13 intact L. multiflorum chromosomes and 1 L. multiflorum
chromosome with a single introgressed F. arundinacea terminal chromosome segment.
The sixth BC(2) winter survivor appeared to be Lolium without any Festuca
introgression capable of detection by GISH. A combined GISH and fluorescence in
situ hybridization analysis with rDNA probes of the most winter-hardy (after 2
winters) and frost-tolerant BC(2) plant revealed the location of an F.
arundinacea introgression on the nonsatellite arm of L. multiflorum chromosome 2,
the same chromosome location reported previously as a site for frost tolerance
genes in the diploid and winter-hardy species Festuca pratensis.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17621586 [PubMed - indexed for MEDLINE]

449: Nat Biotechnol. 2007 Jul;25(7):725-7.

Fatal flaws in agbiotech regulatory policies.

McHughen A.

Department of Botany and Plant Sciences, University of California, Riverside,
California 92521-0124, USA. alanmc@ucr.edu

PMID: 17621292 [PubMed - indexed for MEDLINE]

450: Nat Biotechnol. 2007 Jul;25(7):717-8.

The status of GM rice R&D in China.

Wang Y, Johnston S.

Publication Types:
Letter

PMID: 17621287 [PubMed - indexed for MEDLINE]

451: Curr Opin Allergy Clin Immunol. 2007 Aug;7(4):355-9.

Animal models of anaphylaxis.

Nauta A, Knippels L, Garssen J, Redegeld F.

Numico Research, Wageningen, The Netherlands.

PURPOSE OF REVIEW: In this review we will focus on recent advances in the role of
mast cells in the pathophysiology of insect allergy and the possible mechanisms
of mast cell activation in anaphylaxis. RECENT FINDINGS: Anaphylactic reactions
in the mouse can be induced by several independent pathways involving
immunoglobulin E, immunoglobulin free light chains, or immunoglobulin G. There is
considerable evidence that mast cells play a central role in anaphylactic
reactions to insect stings. Mast cells can be directly activated by components of
insect venom or after allergic sensitization. Of interest is the observation that
mast cells are not only effector cells in insect allergy, but may also play a
protective role in preventing the development of severe anaphylactic responses or
by controlling inflammatory reactions by modulation of antigen-specific T-cell
responses. SUMMARY: The contribution of mast cells in anaphylactic responses to
insect venom may be heterogeneous. On the one hand, activation of mast cells
contributes to the pathology by the release of bioactive and tissue-damaging
mediators. However, mast cell activation may neutralize constituents in insect
venom and defend against the adverse effects of these toxins or they may modulate
inflammation through downregulation of antigen-specific immune responses.

Publication Types:
Review

PMID: 17620830 [PubMed - indexed for MEDLINE]

452: Plant Cell Rep. 2007 Jul;26(7):961-8. Epub 2007 Feb 16.

Transgenic tomatoes express an antigenic polypeptide containing epitopes of the
diphtheria, pertussis and tetanus exotoxins, encoded by a synthetic gene.

Soria-Guerra RE, Rosales-Mendoza S, Mö¡rquez-Mercado C, Lö¨pez-Revilla R,
Castillo-Collazo R, Alpuche-Solö­s AG.

Divisiö¨n de Biologö­a Molecular, Instituto Potosino de Investigaciö¨n Cientö­fica y
Tecnolö¨gica, Camino a la Presa San Josö© 2055, 78216 San Luis Potosö­, S.L.P.,
Mexico.

A current priority of vaccinology is the development of multicomponent vaccines
that protect against several pathogens. The diphtheria-pertussis-tetanus (DPT)
vaccine prevents the symptoms of three serious and often fatal diseases due to
the exotoxins produced by Corynebacterium diphteriae, Bordetella pertussis and
Clostridium tetani. We are attempting to develop an edible DPT multicomponent
vaccine in plants, based on the fusion of protective exotoxin epitopes encoded by
synthetic genes. By means of Agrobacterium mediated transformation we generated
transgenic tomatoes with a plant-optimised synthetic gene encoding a novel
polypeptide containing two adjuvant and six DPT immunoprotective exotoxin
epitopes joined by peptide linkers. In transformed tomato plants, integration of
the synthetic DPT (sDPT) gene detected by PCR was confirmed by Southern blot, and
specific transcripts of the expected molecular size were detected by RT-PCR.
Expression of the putative polypeptide encoded by the sDPT gene was detected by
immunoassay with specific antibodies to the diphtheria, pertussis and tetanus
exotoxins. The sDPT gene is therefore integrated, transcribed and translated as
the expected recombinant sDPT multiepitope polypeptide in transgenic tomatoes
that constitute a potential edible vaccine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17619922 [PubMed - indexed for MEDLINE]

453: ScientificWorldJournal. 2007 Jun 22;7:1047-62.

Cloning, structural characterization, and phylogenetic analysis of flower
MADS-box genes from crocus (Crocus sativus L.).

Tsaftaris AS, Polidoros AN, Pasentsis K, Kalivas A.

Institute of Agrobiotechnology, Center for Research and Technology Hellas,
Thermi, Greece. tsaft@certh.gr

Crocus (Crocus sativus L.) is a crop species cultivated for its flowers and, more
specifically, for its red stigmas. The flower of crocus is bisexual and sterile,
since crocus is a triploid species. Its perianth consists of six petaloid tepals:
three tepals in whorl 1 (outer tepals) and three tepals in whorl 2 (inner
tepals). The androecium consists of three distinct stamens and the gynoecium
consists of a single compound pistil with three carpels, a single three-branched
style, and an inferior ovary. The dry form of the stigmas constitutes the
commercial saffron used as a food additive, in the coloring industry, and in
medicine. In order to uncover and understand the molecular mechanisms controlling
flower development in cultivated crocus and its relative wild progenitor species,
and characterize a number of crocus flower mutants, we have cloned and
characterized different, full-length, cDNA sequences encoding MADS-box
transcription factor proteins involved in flower formation. Here we review the
different methods followed or developed for obtaining these sequences involving
conventional 5 inverted exclamation markö¤ 3 inverted exclamation markö¤ RACE, as
well as newly developed methods from our group, named Rolling Circle
Amplification C RACE (RCA-RACE) and its modification named familyRCA-RACE
(famRCA-RACE). Furthermore, the characteristics of the protein structure and
their common and specific domains for each type of MADS-box transcription factors
in this lower nongrass monocot belonging to the Iridaceae family are described.
Finally, a phylogenetic tree of all the MADS-box sequences available in our lab
is presented and discussed in relation to other data from studies of species of
the Iridaceae group and closely related families from an evolutionary
perspective. The structural and phylogenetic analyses are based on both published
and unpublished data.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17619787 [PubMed - indexed for MEDLINE]

454: Transgenic Res. 2007 Oct;16(5):541-55. Epub 2007 Jul 6.

A multidisciplinary approach directed towards the commercial release of
transgenic herbicide-tolerant rice in Costa Rica.

Espinoza-Esquivel AM, Arrieta-Espinoza G.

Centro de Investigaciö¨n en Biologö­a Celular y Molecular (CIBCM), Ciudad de la
Investigaciö¨n, Universidad de Costa Rica, San Jose, Costa Rica.
amespino@racsa.co.cr

This review discusses a multidisciplinary and multicomponent approach leading to
the development and commercial release of transgenic Costa Rican rice varieties
tolerant to the herbicide gluphosinate ammonium. We describe the field
evaluations of the transgenic lines and their potential environmental impact,
focusing on gene flow, particularly in relation to native wild Oryza species and
weedy rice, based on trials performed in compliance with the national regulatory
requirements of the country. We also present a socio-economic analysis of rice
production in Costa Rica and the economic benefits of genetically modified (GM)
rice as well as an environmental risk-benefit analysis for the deployment of GM
rice. Additionally, food safety evaluation, intellectual property management,
requirements for deregulation, and options for the commercialization of the new
varieties are discussed. We also present results from a national survey aimed at
assessing the level of support for GM crops in Costa Rica as this forms an
integral component of our approach. Taken together, our results demonstrate that
the adoption of these genetically improved rice varieties will provide clear
benefits to Costa Rican rice growers and consumers.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17619158 [PubMed - indexed for MEDLINE]

455: Transgenic Res. 2007 Oct;16(5):581-5. Epub 2007 Jul 6.

Metabolic engineering of carotenoid accumulation by creating a metabolic sink.

Li L, Van Eck J.

USDA-ARS, Plant, Soil and Nutrition Laboratory, Cornell University, Ithaca, NY
14853, USA. ll37@cornell.edu

Carotenoids are highly beneficial for human nutrition and health because they
provide essential nutrients and important antioxidants in our diets. However,
many food crops, especially the major staple crops contain only trace to low
amounts of carotenoids. Although significant progress has been made in developing
food crops rich in carotenoids by altering the expression of carotenoid
biosynthetic genes, in many cases it has proved to be difficult to reach the
desired levels of carotenoid enrichment. The recent identification and
characterization of a novel gene mutation in cauliflower reveals that creating a
metabolic sink to sequester carotenoids is an important mechanism to control
carotenoid accumulation in plants. The successful demonstration of increased
carotenoid accumulation in association with the formation of sink structures in
transgenic crops offers a new and alternative approach to increase carotenoid
content. Manipulation of the formation of metabolic sink along with the catalytic
activity of the pathway may represent a promising strategy for maximally
improving the nutritional quality of food crops.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 17619157 [PubMed - indexed for MEDLINE]

456: Toxicology. 2007 Aug 16;238(1):1-14. Epub 2007 May 24.

Erratum in:
Toxicology. 2008 Jan 14;243(1-2):246. Mitra, Kalyan [added].

Adverse effect of organophosphate compounds, dichlorvos and chlorpyrifos in the
reproductive tissues of transgenic Drosophila melanogaster: 70kDa heat shock
protein as a marker of cellular damage.

Gupta SC, Siddique HR, Mathur N, Mishra RK, Mitra K, Saxena DK, Chowdhuri DK.

Embryotoxicology Section, Industrial Toxicology Research Centre, Lucknow 226001,
India.

The study highlights the adverse effects of organophosphate compounds dichlorvos
and chlorpyrifos on reproduction in Drosophila. Freshly eclosed first instar
larvae of Drosophila melanogaster transgenic for hsp70 (hsp70-lacZ) Bg(9) were
fed on 0.015-150.0ppb dichlorvos and chlorpyrifos mixed food. Virgin flies
eclosing from the normal and contaminated food were pair-mated to examine the
effect of the test chemicals on reproduction of the exposed organisms. Expression
of hsp70, sex peptide (SP or Acp70A), accessory gland protein (Acp36DE) and
tissue damage was examined in reproductive organs of adult fly. Exposed organisms
exhibited a dose-dependent significantly reduced reproductive outcome and males
were found to be more sensitive than females. Hsp70 expression was restricted
only within the testis lobes of male fly while it was not induced in the ovary of
the female. In concurrence with absence of hsp70 expression in the accessory
glands of male fly, tissue damage was evident in them. Acp70A and Acp36DE
expression were found to be significantly downregulated at the higher
concentrations of the test chemicals. The study suggests that (i) dichlorvos is
more deleterious to fly reproduction compared to chlorpyrifos with an adverse
effect on Acp70A and Acp36DE expression required to facilitate normal
reproduction; (ii) hsp70 may be used as a marker of cellular damage against
dichlorvos and chlorpyrifos in Drosophila.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17618723 [PubMed - indexed for MEDLINE]

457: Planta. 2007 Oct;226(5):1243-54. Epub 2007 Jul 6.

Maize Lc transcription factor enhances biosynthesis of anthocyanins, distinct
proanthocyanidins and phenylpropanoids in apple (Malus domestica Borkh.).

Li H, Flachowsky H, Fischer TC, Hanke MV, Forkmann G, Treutter D, Schwab W,
Hoffmann T, Szankowski I.

Institute of Biological Production Systems, Fruit Science Section, Leibniz
University of Hannover, Herrenhaeuser Str. 2, 30419 Hannover, Germany.

Flavonoids are a large family of polyphenolic compounds with manifold functions
in plants. Present in a wide range of vegetables and fruits, flavonoids form an
integral part of the human diet and confer multiple health benefits. Here, we
report on metabolic engineering of the flavonoid biosynthetic pathways in apple
(Malus domestica Borkh.) by overexpression of the maize (Zea mays L.) leaf colour
(Lc) regulatory gene. The Lc gene was transferred into the M. domestica cultivar
Holsteiner Cox via Agrobacterium tumefaciens-mediated transformation which
resulted in enhanced anthocyanin accumulation in regenerated shoots. Five
independent Lc lines were investigated for integration of Lc into the plant
genome by Southern blot and PCR analyses. The Lc-transgenic lines contained one
or two Lc gene copies and showed increased mRNA levels for phenylalanine
ammonia-lyase (PAL), chalcone synthase (CHS), flavanone 3 beta-hydroxylase (FHT),
dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductases (LAR),
anthocyanidin synthase (ANS) and anthocyanidin reductase (ANR). HPLC-DAD and
LC-MS analyses revealed higher levels of the anthocyanin idaein (12-fold), the
flavan 3-ol epicatechin (14-fold), and especially the isomeric catechin
(41-fold), and some distinct dimeric proanthocyanidins (7 to 134-fold) in leaf
tissues of Lc-transgenic lines. The levels of phenylpropanoids and their
derivatives were only slightly increased. Thus, Lc overexpression in Malus
domestica resulted in enhanced biosynthesis of specific flavonoid classes, which
play important roles in both phytopathology and human health.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17618453 [PubMed - indexed for MEDLINE]

458: Plant Cell Environ. 2007 Aug;30(8):994-1005.

Glycinebetaine accumulation is more effective in chloroplasts than in the cytosol
for protecting transgenic tomato plants against abiotic stress.

Park EJ, Jekniä‡ Z, Pino MT, Murata N, Chen TH.

Department of Horticulture, ALS 4017, Oregon State University, Corvallis, OR
97331, USA.

Tomato (Lycopersicon esculentum Mill. cv. Moneymaker) plants were transformed
with a gene for choline oxidase (codA) from Arthrobacter globiformis. The gene
product (CODA) was targeted to the chloroplasts (Chl-codA), cytosol (Cyt-codA) or
both compartments simultaneously (ChlCyt-codA). These three transgenic plant
types accumulated different amounts and proportions of glycinebetaine (GB) in
their chloroplasts and cytosol. Targeting CODA to either the cytosol or both
compartments simultaneously increased total GB content by five- to sixfold over
that measured from the chloroplast targeted lines. Accumulation of GB in codA
transgenic plants was tissue dependent, with the highest levels being recorded in
reproductive organs. Despite accumulating, the lowest amounts of GB, Chl-codA
plants exhibited equal or higher degrees of enhanced tolerance to various abiotic
stresses. This suggests that chloroplastic GB is more effective than cytosolic GB
in protecting plant cells against chilling, high salt and oxidative stresses.
Chloroplastic GB levels were positively correlated with the degree of oxidative
stress tolerance conferred, whereas cytosolic GB showed no such a correlation.
Thus, an increase in total GB content does not necessarily lead to enhanced
stress tolerance, but additional accumulation of chloroplastic GB is likely to
further raise the level of stress tolerance beyond what we have observed.

PMID: 17617827 [PubMed - indexed for MEDLINE]

459: J Exp Bot. 2007;58(11):2851-61. Epub 2007 Jul 7.

Overexpression of HBK3, a class I KNOX homeobox gene, improves the development of
Norway spruce (Picea abies) somatic embryos.

Belmonte MF, Tahir M, Schroeder D, Stasolla C.

Department of Plant Science, University of Manitoba, Winnipeg, R3T 2N2, Manitoba,
Canada.

In order to investigate the effects of HBK3, a spruce gene member of the class I
KNOX family, during somatic embryogenesis, sense (HBK3-S) and antisense (HBK3-A)
Norway spruce (Picea abies) lines were generated. Somatic embryos produced from
these lines were then analysed at morphological and structural levels. Compared
with control, differentiation of immature somatic embryos from pro-embryogenic
masses (PEMs) was accelerated in lines overexpressing HBK3 (HBK3-S). Such
immature embryos showed enlarged embryogenic heads and were able to produce fully
developed cotyledonary embryos at higher frequency. Furthermore, HBK3-S embryos
had enlarged shoot apical meristems (SAMs) and enlarged expression pattern of
PgAGO, a molecular marker gene specific to meristematic cells. Lines in which
HBK3 (HBK3-A) was down-regulated had reduced ability to produce immature somatic
embryos from PEMs and were not able to complete the maturation processes. To
assess the function of HBK3 in comparison with that of angiosperm KNOX genes,
this gene was ectopically expressed in Arabidopsis plants. As observed for
spruce, Arabidopsis embryos overexpressing HBK3 had enlarged meristems and
enlarged expression pattern of SHOOTMERISTEMLESS, a SAM molecular marker gene. In
addition, transformed embryos were able to germinate at a higher rate and the
resulting plants showed a variety of phenotypic aberrations, including abnormal
leaves and reduced apical dominance. Overall, these data confirm the importance
of KNOTTED genes during development and reveal the participation of HBK3 in
conifer embryogeny. Furthermore, the results show redundant functions of this
gene during embryonic growth of spruce and Arabidopsis, but not during
post-embryonic growth.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17617659 [PubMed - indexed for MEDLINE]

460: Gen Comp Endocrinol. 2007 Oct-Dec;154(1-3):128-36. Epub 2007 Jun 3.

Circulating corticosterone levels in breeding blue tits Parus caeruleus differ
between island and mainland populations and between habitats.

Mö¼ller C, Jenni-Eiermann S, Blondel J, Perret P, Caro SP, Lambrechts MM, Jenni L.

Swiss Ornithological Institute, Luzernerstrasse 6, CH-6204 Sempach, Switzerland.
claudia.mueller@vogelwarte.ch

Little is known about whether adaptations to an insular life also involve
adaptations in basal corticosterone levels or in the adrenocortical stress
response, thus being part of a genetically based island syndrome. However,
differences in corticosterone between island and mainland may also be a direct
phenotypic response to differences in environmental conditions or may depend on
individual characteristics of the animal such as body condition or parental
investment. In this paper, we investigated whether insular (Island of Corsica)
and mainland (nearby Southern France) blue tits Parus caeruleus populations
differed in baseline and handling-stress induced corticosterone levels during the
breeding season as a response to biological changes of insular biota. We also
examined whether corticosterone levels of both mainland and insular blue tits
differed between birds living in two different habitats (summergreen and
evergreen oak woods) that differ in food availability and whether individual
characteristics affected corticosterone levels. We found (a) differences in
baseline corticosterone plasma levels between Corsica and the mainland,
independent of regional differences in fat scores, (b) a regional difference in
the relationship between corticosterone levels and brood size, (c) a difference
in the rapidity of onset of the stress response to handling between habitats,
independent of region, and (d) a negative relationship between body fat stores
and baseline corticosterone levels independent of region. Reduced baseline
corticosterone levels on Corsica may be a component of the insular syndrome,
allowing birds to be less aggressive and to enhance parental investment despite
higher breeding densities. We suggest that baseline corticosterone levels are
only elevated if food availability affects directly the parents. However, when
conditions deteriorate unexpectedly (as mimicked by handling stress), food
allocation between parents and offspring needs to be re-adjusted in favor of the
parents, possibly by increased circulating corticosterone levels. The switch to
self-maintenance seems to be modified by the amount of body energy stores.

Publication Types:
Comparative Study

PMID: 17617413 [PubMed - indexed for MEDLINE]

461: Plant J. 2007 Sep;51(5):763-78. Epub 2007 Jul 7.

A protein phosphatase 2A catalytic subunit is a negative regulator of abscisic
acid signalling.

Pernas M, Garcö­a-Casado G, Rojo E, Solano R, Sö¡nchez-Serrano JJ.

Centro Nacional de Biotecnologö­a CSIC, Campus de Cantoblanco UAM, 28049, Madrid,
Spain.

The key regulatory role of abscisic acid (ABA) in many physiological processes in
plants is well established. However, compared with other plant hormones, the
molecular mechanisms underlying ABA signalling are poorly characterized. In this
work, a specific catalytic subunit of protein phosphatase 2A (PP2Ac-2) has been
identified as a component of the signalling pathway that represses responses to
ABA. A loss-of-function pp2ac-2 mutant is hypersensitive to ABA. Moreover,
pp2ac-2 plants have altered responses in developmental and environmental
processes that are mediated by ABA, such as primary and lateral root development,
seed germination and responses to drought and high salt and sugar stresses.
Conversely, transgenic plants overexpressing PP2Ac-2 are less sensitive to ABA
than wild type, a phenotype that is manifested in all the above-mentioned
physiological processes. DNA microarray hybridization experiments reveal that
PP2Ac-2 is negatively involved in ABA responses through regulation of
ABA-dependent gene expression. Moreover, the results obtained indicate that ABA
antagonistically regulates PP2Ac-2 expression and PP2Ac-2 activity thus allowing
plant sensitivity to the hormone to be reset after induction. Phenotypic, genetic
and gene expression data strongly suggest that PP2Ac-2 is a negative regulator of
the ABA pathway. Activity of protein phosphatase 2A thus emerges as a key element
in the control of ABA signalling.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17617176 [PubMed - indexed for MEDLINE]

462: J Exp Bot. 2007;58(11):2799-810. Epub 2007 Jul 5.

Molecular cloning of a bifunctional beta-xylosidase/alpha-L-arabinosidase from
alfalfa roots: heterologous expression in Medicago truncatula and substrate
specificity of the purified enzyme.

Xiong JS, Balland-Vanney M, Xie ZP, Schultze M, Kondorosi A, Kondorosi E,
Staehelin C.

State Key Laboratory of Biocontrol, 135, Xingangxi Road, School of Life Sciences,
SunYat-Sen (Zhongshan) University, Guangzhou 510275, China.

Glycoside hydrolases are often members of a multigene family, suggesting
individual roles for each isoenzyme. Various extracellular glycoside hydrolases
have an important but poorly understood function in remodelling the cell wall
during plant growth. Here, MsXyl1, a concanavalin A-binding protein from alfalfa
(Medicago sativa L.) belonging to the glycoside hydrolase family 3
(beta-D-xylosidase branch) is characterized. Transcripts of MsXyl1 were detected
in roots (particularly root tips), root nodules, and flowers. MsXyl1 under the
control of the CaMV 35S promoter was expressed in the model legume Medicago
truncatula (Gaertner). Concanavalin A-binding proteins from the transgenic plants
exhibited 5-8-fold increased activities towards three p-nitrophenyl (PNP)
glycosides, namely PNP-beta-D-xyloside, PNP-alpha-L-arabinofuranoside, and
PNP-alpha-L-arabinopyranoside. An antiserum raised against a synthetic peptide
recognized MsXyl1, which was processed to a 65 kDa form. To characterize the
substrate specificity of MsXyl1, the recombinant protein was purified from
transgenic M. truncatula leaves by concanavalin A and anion chromatography.
MsXyl1cleaved beta-1,4-linked D-xylo-oligosaccharides and alpha-1,5-linked
L-arabino-oligosaccharides. Arabinoxylan (from wheat) and arabinan (from sugar
beet) were substrates for MsXyl1, whereas xylan (from oat spelts) was resistant
to degradation. Furthermore, MsXyl1 released xylose and arabinose from cell wall
polysaccharides isolated from alfalfa roots. These data suggest that MsXyl1 is a
multifunctional
beta-xylosidase/alpha-L-arabinofuranosidase/alpha-L-arabinopyranosidase
implicated in cell wall turnover of arabinose and xylose, particularly in rapidly
growing root tips. Moreover, the findings of this study demonstrate that stable
transgenic M. truncatula plants serve as an excellent expression system for
purification and characterization of proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17615411 [PubMed - indexed for MEDLINE]

463: Plant Biotechnol J. 2007 Sep;5(5):646-56. Epub 2007 Jul 5.

Expression of barley HvCBF4 enhances tolerance to abiotic stress in transgenic
rice.

Oh SJ, Kwon CW, Choi DW, Song SI, Kim JK.

School of Biotechnology and Environmental Engineering, Myongji University, Yongin
449-728, South Korea.

C-repeat/dehydration-responsive element binding factors (CBF/DREBs) are a family
of transcription factors that regulate freezing tolerance in Arabidopsis. As a
step towards understanding the stress response of monocotyledonous plants, we
isolated a barley gene HvCBF4 whose expression is induced by low-temperature
stress. Transgenic over-expression of HvCBF4 in rice resulted in an increase in
tolerance to drought, high-salinity and low-temperature stresses without stunting
growth. Interestingly, under low-temperature conditions, the maximum
photochemical efficiency of photosystem II in the dark-adapted state (F(v)/F(m),
where F(v) is the variable fluorescence and F(m) is the maximum fluorescence) in
HvCBF4 plants was higher by 20% and 10% than that in non-transgenic and
CBF3/DREB1A plants, respectively. Using the 60K Rice Whole Genome microarray, 15
rice genes were identified that were activated by HvCBF4. When compared with 12
target rice genes of CBF3/DREB1A, five genes were common to both HvCBF4 and
CBF3/DREB1A, and 10 and seven genes were specific to HvCBF4 and CBF3/DREB1A,
respectively. Interestingly, HvCBF4 did not activate Dip1 and Lip5, two important
target genes of CBF3/DREB1A, in transgenic rice under normal growth conditions,
but their expression was enhanced by HvCBF4 under low-temperature conditions. Our
results suggest that CBF/DREBs of barley act differently from those of
Arabidopsis in transgenic rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17614953 [PubMed - indexed for MEDLINE]

464: Plant Biotechnol J. 2007 Nov;5(6):709-19. Epub 2007 Jul 5.

Subcellular targeting is a key condition for high-level accumulation of cellulase
protein in transgenic maize seed.

Hood EE, Love R, Lane J, Bray J, Clough R, Pappu K, Drees C, Hood KR, Yoon S,
Ahmad A, Howard JA.

Arkansas State University, PO Box 2760, State University, AR 72467, USA.
ehood@astate.edu

Ethanol from lignocellulosic biomass is being pursued as an alternative to
petroleum-based transportation fuels. To succeed in this endeavour, efficient
digestion of cellulose into monomeric sugar streams is a key step. Current
production systems for cellulase enzymes, i.e. fungi and bacteria, cannot meet
the cost and huge volume requirements of this commodity-based industry.
Transgenic maize (Zea mays L.) seed containing cellulase protein in embryo
tissue, with protein localized to the endoplasmic reticulum, cell wall or
vacuole, allows the recovery of commercial amounts of enzyme. E1 cellulase, an
endo-beta-1,4-glucanase from Acidothermus cellulolyticus, was recovered at levels
greater than 16% total soluble protein (TSP) in single seed. More significantly,
cellobiohydrolase I (CBH I), an exocellulase from Trichoderma reesei, also
accumulated to levels greater than 16% TSP in single seed, nearly 1000-fold
higher than the expression in any other plant reported in the literature. The
catalytic domain was the dominant form of E1 that was detected in the endoplasmic
reticulum and vacuole, whereas CBH I holoenzyme was present in the cell wall.
With one exception, individual transgenic events contained single inserts.
Recovery of high levels of enzyme in T2 ears demonstrated that expression is
likely to be stable over multiple generations. The enzymes were active in
cleaving soluble substrate.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17614952 [PubMed - indexed for MEDLINE]

465: Plant Mol Biol. 2007 Sep;65(1-2):93-106. Epub 2007 Jul 5.

Leaf rust resistance gene Lr1, isolated from bread wheat (Triticum aestivum L.)
is a member of the large psr567 gene family.

Cloutier S, McCallum BD, Loutre C, Banks TW, Wicker T, Feuillet C, Keller B,
Jordan MC.

Cereal Research Centre, Agriculture and Agri-Food Canada, R3T 2M9, Winnipeg, MB,
Canada. scloutier@agr.gc.ca

In hexaploid wheat, leaf rust resistance gene Lr1 is located at the distal end of
the long arm of chromosome 5D. To clone this gene, an F(1)-derived doubled
haploid population and a recombinant inbred line population from a cross between
the susceptible cultivar AC Karma and the resistant line 87E03-S2B1 were
phenotyped for resistance to Puccinia triticina race 1-1 BBB that carries the
avirulence gene Avr1. A high-resolution genetic map of the Lr1 locus was
constructed using microsatellite, resistance gene analog (RGA), BAC end (BE), and
low pass (LP) markers. A physical map of the locus was constructed by screening a
hexaploid wheat BAC library from cultivar Glenlea that is known to have Lr1. The
locus comprised three RGAs from a gene family related to RFLP marker Xpsr567.
Markers specific to each paralog were developed. Lr1 segregated with RGA567-5
while recombinants were observed for the other two RGAs. Transformation of the
susceptible cultivar Fielder with RGA567-5 demonstrated that it corresponds to
the Lr1 resistance gene. In addition, the candidate gene was also confirmed by
virus-induced gene silencing. Twenty T (1) lines from resistant transgenic line T
(0)-938 segregated for resistance, partial resistance and susceptibility to Avr1
corresponding to a 1:2:1 ratio for a single hemizygous insertion. Transgene
presence and expression correlated with the phenotype. The resistance phenotype
expressed by Lr1 seemed therefore to be dependant on the zygosity status. T
(3)-938 sister lines with and without the transgene were further tested with 16
virulent and avirulent rust isolates. Rust reactions were all as expected for Lr1
thereby providing additional evidence toward the Lr1 identity of RGA567-5.
Sequence analysis of Lr1 indicated that it is not related to the previously
isolated Lr10 and Lr21 genes and unlike these genes, it is part of a large gene
family.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17611798 [PubMed - indexed for MEDLINE]

466: Theor Appl Genet. 2007 Aug;115(4):561-70. Epub 2007 Jul 4.

Analysis of the chromosome 2(2H) region of barley associated with the correlated
traits Fusarium head blight resistance and heading date.

Nduulu LM, Mesfin A, Muehlbauer GJ, Smith KP.

Department of Agronomy and Plant Genetics, University of Minnesota, Rm 411
Borlaug Hall, 1991 Upper Buford Circle, St. Paul, MN 55108, USA.

Fusarium head blight (FHB) is a major disease of barley (Hordeum vulgare L.) that
results in reduced grain yield and quality through the accumulation of the
mycotoxin deoxynivalenol (DON). Coincident QTL for FHB severity, DON
concentration, and heading date (HD) map to a region of chromosome 2(2H)
designated Qrgz-2H-8. It is unclear whether disease resistance at this locus is
due to a pleiotropic effect of late HD by delaying the host exposure to the
pathogen or a tightly linked resistance gene. The objectives of this study were
to develop a set of near isogenic lines (NILs) for the Qrgz-2H-8 region and to
genetically dissect the QTL region containing the coincident traits. Two NIL
populations were developed consisting of F(2)- and F(4)-derived recombinants from
a cross between a BC(5) line carrying the donor parent (Chevron) alleles in the
Qrgz-2H-8 region and the recurrent parent M69. Analysis of field and marker data
from these NILs revealed that the Chevron alleles conditioning FHB resistance,
late HD, and low DON concentration were successfully introgressed into the BC(5)
parent line and were segregating among NILs. QTL analysis of the F(4)-derived
population showed that the HD QTL is adjacent to the FHB QTL. Furthermore, a
single NIL was identified that was similar to the resistant BC(5) parent for FHB
severity and the early flowering parent M69 for HD. These results indicate that
the relationship between FHB and HD at the Qrgz-2H-8 region is likely due to
tight linkage rather than pleiotropy.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17609926 [PubMed - indexed for MEDLINE]

467: Biotechnol Lett. 2007 Nov;29(11):1781-7. Epub 2007 Jul 4.

Phytase expression in transgenic soybeans: stable transformation with a
vector-less construct.

Gao XR, Wang GK, Su Q, Wang Y, An LJ.

Department of Bioscience and Biotechnology, Dalian University of Technology,
Dalian, 116024, P.R. China. gxiaorong@yahoo.com

A minimal linear gene cassette (35S-phytase gene-nos) with T-DNA borders was
acquired by PCR and directly introduced into soybean through the pollen tube
pathway. A total of 13% of T(1 )plants were positive for phyA by specific PCR.
Southern blot analyses showed that phyA insertions were harbored stably in T(2)
progeny. Phytase expression level increased 2.5-fold over a 6-week period; its
highest activity was 150 U/mg protein, compared to 56 U/mg protein in
untransformed controls. Activity of phytase increased to 125 FTU/kg in T(3)
transgenic seeds as compared to 64 FTU/kg in wild-type plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17609861 [PubMed - indexed for MEDLINE]

468: Plant Cell Physiol. 2007 Aug;48(8):1108-20. Epub 2007 Jul 2.

A rice dihydrosphingosine C4 hydroxylase (DSH1) gene, which is abundantly
expressed in the stigmas, vascular cells and apical meristem, may be involved in
fertility.

Imamura T, Kusano H, Kajigaya Y, Ichikawa M, Shimada H.

Department of Biological Science and Technology, Tokyo University of Science,
2641 Yamazaki, Noda, Chiba, 278-8510, Japan.

Dihydrosphingosine C4 hydroxylase is a key enzyme in the biosynthesis of
phytosphingosine, a major constituent of sphingolipids in plants and yeasts. The
rice genome contains five homologue genes for dihydrosphingosine C4 hydroxylase,
DSH1-DSH5, whose gene products show high degrees of homology to the yeast
counterpart, SUR2. Among them, expression of DSH1, DSH2 and DSH4 was detected,
and DSH1 and DSH4 complement the yeast sur2 mutation. The DSH1 gene was
specifically and abundantly expressed in vascular bundles and apical meristems.
In particular, very strong expression was detected in the stigmas of flowers.
Repression of DSH1 expression by the antisense gene or RNA interference (RNAi)
resulted in a severe reduction of fertility. In the transformants in which DSH1
expression was suppressed, significantly increased expression of DSH2 was found
in leaves but not in pistils, suggesting that there was tissue-specific
correlation between DSH1 and DSH2 expression. Our results indicate that the
product of DSH1 may be involved in plant viability or reproductive processes, and
that the phenotype of sterility is apparently caused by loss of function of DSH1
in the stigma. It is also suggested that there is a complex mechanism controlling
the tissue-specific expression of the DSH1 gene.

PMID: 17609219 [PubMed - indexed for MEDLINE]

469: J Agric Food Chem. 2007 Jul 25;55(15):5942-7. Epub 2007 Jul 4.

Indicated detection of two unapproved transgenic rice lines contaminating
vermicelli products.

Akiyama H, Sasaki N, Sakata K, Ohmori K, Toyota A, Kikuchi Y, Watanabe T, Furui
S, Kitta K, Maitani T.

National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo
158-8501, Japan, Tokyo University of Agriculture and Technology, 2-24-16,
Naka-cho, Koganei, Tokyo 184-8588, Japan. akiyama@nihs.go.jp

We analyzed the DNA fragments extracted from four rice vermicelli products. The
Bacillus thuringiensis (Bt) rice line, which has a construct similar to the GM
Shanyou 63 line, was detected in some vermicelli products by identification of
the junction region sequence between rice Act1 promoter and the Cry1Ac gene, and
that between Cry1Ac and nos. In addition, we also detected a different Bt rice
line by means of the junction region sequence between the maize ubiquitin
promoter and cry1Ab gene and that between the cauliflower mosaic virus 35S
promoter and the hygromycin phosphotransferase in some vermicelli products.
Accordingly, we for the first time have detected the two transgenic Bt rice lines
contaminating rice vermicelli samples. Furthermore, we developed a duplex
real-time polymerase chain reaction (PCR) method for the simultaneous detection
of both Bt rice lines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17608495 [PubMed - indexed for MEDLINE]

470: J Agric Food Chem. 2007 Jul 25;55(15):6160-8. Epub 2007 Jul 3.

Chemical composition of glyphosate-tolerant soybean 40-3-2 grown in Europe
remains equivalent with that of conventional soybean (Glycine max L.).

Harrigan GG, Ridley WP, Riordan SG, Nemeth MA, Sorbet R, Trujillo WA, Breeze ML,
Schneider RW.

Product Safety Center and Regulatory Affairs, Monsanto Company, 800 North
Lindbergh Boulevard, St. Louis, Missouri 63167, USA.
george.g.harrigan@monsanto.com

The composition of glyphosate-tolerant (Roundup Ready) soybean 40-3-2 was
compared with that of conventional soybean grown in Romania in 2005 as part of a
comparative safety assessment program. Samples were collected from replicated
field trials, and compositional analyses were performed to measure proximates
(moisture, fat, ash, protein, and carbohydrates by calculation), fiber, amino
acids, fatty acids, isoflavones, raffinose, stachyose, phytic acid, trypsin
inhibitor, and lectin in grain as well as proximates and fiber in forage. The
mean values for all biochemical components assessed for Roundup Ready soybean
40-30-2 were similar to those of the conventional control and were within the
published range observed for commercial soybean. The compositional profile of
Roundup Ready soybean 40-3-2 was also compared to that of conventional soybean
varieties grown in Romania by calculating a 99% tolerance interval to describe
compositional variability in the population of traditional soybean varieties
already on the marketplace. These comparisons, together with the history of the
safe use of soybean as a common component of animal feed and human food, lead to
the conclusion that Roundup Ready soybean 40-3-2 is compositionally equivalent to
and as safe and nutritious as conventional soybean varieties grown commercially.

Publication Types:
Comparative Study

PMID: 17608426 [PubMed - indexed for MEDLINE]

471: Electrophoresis. 2007 Jul;28(13):2314-23.

Characterization and differentiation of diverse transgenic and nontransgenic
soybean varieties from CE protein profiles.

Garcö­a-Ruiz C, Garcö­a MC, Cifuentes A, Marina ML.

Department of Analytical Chemistry, Faculty of Chemistry, University of Alcalö¡,
Ctra. Madrid-Barcelona, Alcalö¡ de Henares, Madrid, Spain.

Nowadays, soybeans are commercialized in a wide variety of colors and tones.
Moreover, some pigmented seeds are being commercialized as soybeans while, on
other occasions, these seeds are labeled as mung beans, azuki beans or soybean
frijoles generating confusion on their identity. In this work, CE has been
applied for the first time for the characterization and differentiation of
different pigmented beans commercialized as soybeans. Other seeds commercialized
as azuki, mung green soybeans or soybean frijoles were also analyzed. Borate
buffer (at pH 8.5) containing 20% v/v ACN was used as the separation media and
solution containing ACN/water (75:25 v/v) with 0.3% v/v acetic acid was used to
solubilize the proteins from the samples. A 50 cm bare fused-silica capillary was
employed for obtaining adequate separations in about 12 min. The CE protein
pattern observed for yellow soybeans was different from that corresponding to
green and red soybeans. The seeds commercialized as black soybean presented
electropherograms identical or similar to those yielded by the yellow seeds with
the exception of the sample labeled as black soybeans frijoles that presented a
totally different pattern. In addition, CE protein profiles obtained for azuki
and mung green soybeans were very similar to those corresponding to red soybeans
and green soybeans, respectively. Finally, the CE method was also applied to
differentiate transgenic and nontransgenic soybean varieties. Discriminant
analysis, using several protein peak areas as variable, was used to successfully
classify these samples.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17607812 [PubMed - indexed for MEDLINE]

472: Neotrop Entomol. 2007 Mar-Apr;36(2):288-93.

[Mites associated with soybean crop in Rio Grande do Sul State, Brazil]

[Article in Portuguese]

Guedes JV, Navia D, Lofego AC, Dequech ST.

Depto. Defesa Fitossanitö¡ria, Univ. Federal de Santa Maria, Santa Maria, RS.

During the last growing seasons, high infestations of phytophagous mites were
observed in the State of Rio Grande do Sul, Brazil, becoming necessary to apply
pesticides for their control. The objective of this study was to identify
phytophagous and predatory mite species associated with soybean in ten counties
of that state, during the 2002/03 and 2003/04 growing seasons, in five soybean
cultivars (A 6001 RG, A 7001 RG, A 8000 RG, A 8100 RG, Anta 82), all genetically
modified. In samples of soybean leaves four phytophagous mite species
(Mononychellus planki (McGregor), Polyphagotarsonemus latus (Banks), Tetranychus
desertorum Banks and Tetranychus gigas Pritchard & Baker) and two predatory mite
species (Phytoseiulus fragariae Denmark & Schicha and Typhlodromalus aripo De
Leon) were found. T. desertorum was found for the first time associated with
soybean in the country. Phytoseiulus fragariae and T. aripo are reported for the
first time on soybean. The potential of phytoseid mites as biological control
agents in soybean crop was discussed. Among the hypotheses to explain the
increasing infestation of soybean fields with phytophagous mites area are the
progressively larger cultivated area, the dry spells observed in the last few
years in the growing season, changes in soybean cropping system that led to
increased use of pesticides and utilization of new soybean cultivars with
morphological or biochemicals characteristics that favour the development of
these mite populations.

Publication Types:
English Abstract

PMID: 17607464 [PubMed - indexed for MEDLINE]

473: Public Health Nutr. 2008 Jan;11(1):8-16. Epub 2007 Jul 3.

Predictors of Australian consumers' intentions to consume conventional and novel
sources of long-chain omega-3 fatty acids.

Cox DN, Evans G, Lease HJ.

CSIRO Food Futures National Research Flagship and CSIRO Human Nutrition, Adelaide
BC, South Australia 5000, Australia. david.cox@csiro.au

OBJECTIVES: To elicit predictors of variation in likelihood to purchase foods
rich in long-chain omega-3 fatty acids. DESIGN, SETTING AND SUBJECTS: Responses
from a community sample (n = 220) were elicited using a computer-administered
questionnaire based on an adaptation of Protection Motivation Theory including
measures of perceived risk and vulnerability to coronary heart disease (CHD).
Other measures included health status, body mass index (BMI), perceived
risk/benefits of novel technologies and sociodemographics. Descriptions of model
products were presented, including farmed fish fed fishmeal (FFFF); farmed fish
fed genetically modified (GM) oilseed (FFFGM); bread, milk and supplements
containing fish oil (SFO) or GM oilseed. It was hypothesised that perceived
vulnerability to CHD would enhance acceptance of GM products (H1). Furthermore,
information describing the benefits of LCO3FA, limitations to fish supply and
potential alternatives was given to a treatment group (50%) and hypothesised to
have a positive effect on the acceptance of GM products (H2). RESULTS: No
evidence was found to support H1 or H2. FFFF was most likely to be purchased (P <
0.01), followed by SFO and FFFGM. Multivariate regression analysis identified
significant (P < 0.05) predictors (standardised beta) for likelihood to purchase
FFFF: self-efficacy 0.56; behaviour (product) efficacy 0.19; belief that fishmeal
is unnatural -0.14 (R2 = 0.44) and for FFFGM: self-efficacy 0.65; perceived
severity of CHD 0.15; BMI -0.13; significant other has/had arthritis 0.11; belief
that GM oilseed is unnatural 0.11 (R2 = 0.49). CONCLUSIONS: Self-efficacy
(confidence to consume) was the most important predictor of likelihood to
purchase all products.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17605836 [PubMed - indexed for MEDLINE]

474: J Plant Physiol. 2007 Oct;164(10):1367-76. Epub 2007 Jul 2.

Stress-induced synthesis of proline confers tolerance to water deficit in
transgenic wheat.

Vendruscolo EC, Schuster I, Pileggi M, Scapim CA, Molinari HB, Marur CJ, Vieira
LG.

Universidade Federal do Paranö¡-Campus Palotina, Rua Pioneiro, 2153, CEP 85950-000
Palotina-Pr, Brazil. vendruscolo@ufpr.br

Water deficit is one of the main abiotic factors that affect spring wheat planted
in subtropical regions. Accumulation of proline appears to be a promising
approach to maintain the productivity of plants under stress condition. However,
morphological alterations and growth reduction are observed in transgenic plants
carrying genes coding for osmoprotectants controlled by constitutive promoters.
We report here the effects of water deficit on wheat plants transformed with the
Vigna aconitifolia Delta(1)-pyrroline-5-carboxylate synthetase (P5CS) cDNA that
encodes the key regulatory enzyme in proline biosynthesis, under the control of a
stress-induced promoter complex-AIPC. Transgenic wheat plants submitted to 15
days of water shortage presented a distinct response. We have found that drought
resulted in the accumulation of proline. The tolerance to water deficit observed
in transgenic plants was mainly due to protection mechanisms against oxidative
stress and not caused by osmotic adjustment.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17604875 [PubMed - indexed for MEDLINE]

475: Mol Plant Microbe Interact. 2007 Jul;20(7):832-42.

Sucrose-mediated priming of plant defense responses and broad-spectrum disease
resistance by overexpression of the maize pathogenesis-related PRms protein in
rice plants.

Gö¨mez-Ariza J, Campo S, Rufat M, Estopö  M, Messeguer J, San Segundo B, Coca M.

Departamento de Genö©tica Molecular, Laboratorio de Genö©tica Molecular Vegetal,
Consorcio CSIC-IRTA, Jordi Girona 18, 08034 Barcelona, Spain.

Expression of pathogenesis-related (PR) genes is part of the plant's natural
defense response against pathogen attack. The PRms gene encodes a
fungal-inducible PR protein from maize. Here, we demonstrate that expression of
PRms in transgenic rice confers broad-spectrum protection against pathogens,
including fungal (Magnaporthe oryzae, Fusarium verticillioides, and
Helminthosporium oryzae) and bacterial (Erwinia chrysanthemi) pathogens. The
PRms-mediated disease resistance in rice plants is associated with an enhanced
capacity to express and activate the natural plant defense mechanisms. Thus, PRms
rice plants display a basal level of expression of endogenous defense genes in
the absence of the pathogen. PRms plants also exhibit stronger and quicker
defense responses during pathogen infection. We also have found that sucrose
accumulates at higher levels in leaves of PRms plants. Sucrose responsiveness of
rice defense genes correlates with the pathogen-responsive priming of their
expression in PRms rice plants. Moreover, pretreatment of rice plants with
sucrose enhances resistance to M. oryzae infection. Together, these results
support a sucrose-mediated priming of defense responses in PRms rice plants which
results in broad-spectrum disease resistance.

PMID: 17601170 [PubMed - indexed for MEDLINE]

476: Mol Plant Microbe Interact. 2007 Jul;20(7):769-80.

Promoters of orthologous Glycine max and Lotus japonicus nodulation
autoregulation genes interchangeably drive phloem-specific expression in
transgenic plants.

Nontachaiyapoom S, Scott PT, Men AE, Kinkema M, Schenk PM, Gresshoff PM.

Australian Research Council Centre of Excellence for Integrative Legume Research,
University of Queensland, St. Lucia, QLD 4072, Australia.

The nodule autoregulation receptor kinase (GmNARK) of soybean (Glycine max) is
essential for the systemic autoregulation of nodulation. Based on quantitative
reverse-transcriptase polymerase chain reaction, GmNARK is ex-pressed to varying
levels throughout the plant; the transcript was detected at high levels in mature
leaves and roots but to a lesser extent in young leaves, shoot tips, and nodules.
The transcript level was not significantly affected by Bradyrhizobium japonicum
during the first week following inoculation. In addition, the activities of the
promoters of GmNARK and Lotus japonicus HARI, driving a beta-glucuronidase
(GUSPlus) reporter gene, were examined in stably transformed L. japonicus and
transgenic hairy roots of soybean. Histochemical GUS activity in L. japonicus
plants carrying either a 1.7-kb GmNARKpr::GUS or 2.0-kb LjHAR1pr::GUS construct
was clearly localized to living cells within vascular bundles, especially phloem
cells in leaves, stems, roots, and nodules. Phloem-specific expression also was
detected in soybean hairy roots carrying these constructs. Our study suggests
that regulatory elements required for the transcription of these orthologous
genes are conserved. Moreover, rapid amplification of 5' cDNA ends (5' rapid
amplification of cDNA ends) revealed two major transcripts of GmNARK potentially
originating from two TATA boxes. Further analysis of the GmNARK promoter has
confirmed that these two TATA boxes are functional. Deletion analysis also
located a region controlling phloem-specific expression to a DNA sequence between
908 bp and 1.7 kb upstream of the translation start site of GmNARK.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17601165 [PubMed - indexed for MEDLINE]

477: Plant Physiol. 2007 Aug;144(4):1968-77. Epub 2007 Jun 28.

A higher plant delta8 sphingolipid desaturase with a preference for (Z)-isomer
formation confers aluminum tolerance to yeast and plants.

Ryan PR, Liu Q, Sperling P, Dong B, Franke S, Delhaize E.

Commonwealth Scientific and Industrial Research Organization, Plant Industry,
Canberra, Australian Capital Territory 2601, Australia. peter.ryan@csiro.au

Three plant cDNA libraries were expressed in yeast (Saccharomyces cerevisiae) and
screened on agar plates containing toxic concentrations of aluminum. Nine cDNAs
were isolated that enhanced the aluminum tolerance of yeast. These cDNAs were
constitutively expressed in Arabidopsis (Arabidopsis thaliana) and one cDNA from
the roots of Stylosanthes hamata, designated S851, conferred greater aluminum
tolerance to the transgenic seedlings. The protein predicted to be encoded by
S851 showed an equally high similarity to Delta6 fatty acyl lipid desaturases and
Delta8 sphingolipid desaturases. We expressed other known Delta6 desaturase and
Delta8 desaturase genes in yeast and showed that a Delta6 fatty acyl desaturase
from Echium plantagineum did not confer aluminum tolerance, whereas a Delta8
sphingobase desaturase from Arabidopsis did confer aluminum tolerance. Analysis
of the fatty acids and sphingobases of the transgenic yeast and plant cells
demonstrated that S851 encodes a Delta8 sphingobase desaturase, which leads to
the accumulation of 8(Z/E)-C(18)-phytosphingenine and
8(Z/E)-C(20)-phytopshingenine in yeast and to the accumulation of
8(Z/E)-C(18)-phytosphingenine in the leaves and roots of Arabidopsis plants. The
newly formed 8(Z/E)-C(18)-phytosphingenine in transgenic yeast accounted for 3
mol% of the total sphingobases with a 8(Z):8(E)-isomer ratio of approximately
4:1. The accumulation of 8(Z)-C(18)-phytosphingenine in transgenic Arabidopsis
shifted the ratio of the 8(Z):8(E) isomers from 1:4 in wild-type plants to 1:1 in
transgenic plants. These results indicate that S851 encodes the first Delta8
sphingolipid desaturase to be identified in higher plants with a preference for
the 8(Z)-isomer. They further demonstrate that changes in the sphingolipid
composition of cell membranes can protect plants from aluminum stress.

PMID: 17600137 [PubMed - indexed for MEDLINE]

478: J Econ Entomol. 2007 Jun;100(3):921-31.

Susceptibility of bollworm and tobacco budworm (Lepidoptera: Noctuidae) to
Cry2Ab2 insecticidal protein.

Ali MI, Luttrell RG.

Department of Entomology, University of Arkansas, Fayetteville, AR 72701, USA.
miali@uark.edu

Susceptibilities of 82 bollworm, Helicoverpa zea (Boddie), and 44 tobacco
budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae), populations to
Cry2Ab2 protein were measured in diet incorporated assays at the University of
Arkansas from 2002 to 2005. Resulting data were used to calculate overall (pooled
data) estimates of species susceptibility for future benchmarks of resistance.
Variabilities among populations also were studied by comparing regressions for
individual populations and calculating mean susceptibilities for different
subgroups of the colonies studied. Individual lethal concentration (LC50)
estimates for nine laboratory, seven laboratory-cross, and 28 field populations
of H. virescens varied up to 48-fold when adjusted for the response of the most
susceptible laboratory colony studied. Mean susceptibilities of all laboratory,
laboratory-cross, or field colonies varied only two-fold. When grouped by host
plants, populations collected on tobacco, Nicotiana tabacum (L.), seemed to be
less susceptible than those collected on other host plants. Individual LC50
values for 82 laboratory, laboratory-cross and field populations of H. zea varied
up to 37-fold. Mean LC50 values of all laboratory, laboratory-cross, or field
populations varied only three-fold. Susceptibilities of populations from Bollgard
cotton were up to four-fold less than those from Bacillus thuringiensis corn, Zea
mays L. Field populations collected during late season were generally less
susceptible than those collected early in the season. Across the two species, H.
zea was less sensitive to Cry2Ab2 than H. virescens. Both species seem to be less
sensitive to Cry2Ab2 than to CrylAc.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17598557 [PubMed - indexed for MEDLINE]

479: Nature. 2007 Jun 28;447(7148):1042.

Uganda hosts banana trial.

Dauwers A.

Publication Types:
News

PMID: 17597729 [PubMed - indexed for MEDLINE]

480: Biosci Rep. 2007 Oct;27(4-5):225-34.

Overexpression of the rFCA RNA recognition motif affects morphologies
modifications in rice (Oryza sativa L.).

Hong F, Attia K, Wei C, Li K, He G, Su W, Zhang Q, Qian X, Yang J.

Institute of Genetics, State Key Laboratory of Genetic Engineering, School of
Life Sciences, Fudan University, Shanghai, PR China.

RNA recognition motifs as important regulators of gene expression are highly
conserved in animals and plants. The FCA floral promotion gene in Arabidopsis
encodes a protein, containing two RNA recognition motifs (RRM) and a WW protein
interaction domain. Here we isolated FCA cDNA from rice. FCA in rice (rFCA) was
homologous to FCA-gamma of Arabidopsis and contained conserved domains. To
investigate the function of RRM domain, fragment RRM1 and RRM2 of rFCA were
introduced into rice subspecies Oryza sativa L. subsp. Indica var. 9311 and
another rice subspecies Oryza sativa L. subsp. Japonica var. zhonghua11
transformation. Two transgenic lines exhibited similar phenotypes, flowering time
delay, seed size and cell volume of transgenic plants was increased. These
results showed that constitutive expression of RRMs could regulate cellular size.
The patterns of overexpression of two RRM domains and their similar morphologies
indicate they may play a same role.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17597396 [PubMed - indexed for MEDLINE]

481: Plant Cell Physiol. 2007 Aug;48(8):1098-107. Epub 2007 Jun 27.

Transactivation of protein expression by rice HSP101 in planta and using Hsp101
as a selection marker for transformation.

Chang CC, Huang PS, Lin HR, Lu CH.

Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan 701.
chingcc@mail.ncku.edu.tw

Plant HSP101 has dual activities, first, in conferring thermotolerance, and
secondly, in serving as a translational activator. In this study, we introduced
Oryza sativa Hsp101 (osHsp101) cDNA into tobacco by Agrobacterium-mediated
transformation. Stable integration and expression of the transgene into the
tobacco genome was demonstrated by Southern and Western blot analysis.
Overexpression of osHSP101 had no noticeable effect on growth or development of
the transgenic plants. Homozygous T(2) transgenic plants with overexpressed
osHSP101 survived heat treatment better than untransformed control plants. In
addition, taking advantage of conferring basal thermotolerance by plant HSP101,
we were able to demonstrate the feasibility of using osHsp101 as a selection
marker and select the transformants under high temperature in tobacco leaf disc
transformation mediated by Agrobacterium. Furthermore, transgenic tobacco plants
with overexpressed osHSP101 were able to enhance luciferase expression up to
2.9-fold more than untransformed plants in the progeny of reciprocally crossed
with omega-luciferase reporter lines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17597080 [PubMed - indexed for MEDLINE]

482: Plant Biotechnol J. 2007 Sep;5(5):664-74. Epub 2007 Jun 26.

A rice promoter containing both novel positive and negative cis-elements for
regulation of green tissue-specific gene expression in transgenic plants.

Cai M, Wei J, Li X, Xu C, Wang S.

National Key Laboratory of Crop Genetic Improvement, National Centre of Plant
Gene Research (Wuhan), Huazhong Agricultural University, Wuhan 430070, China.

The tissue-specific expression of transgenes is essential in plant breeding
programmes to avoid the fitness costs caused by constitutive expression of a
target gene. However, knowledge on the molecular mechanisms of tissue-specific
gene expression and practicable tissue-specific promoters is limited. In this
study, we identified the cis-acting elements of a tissue-specific promoter from
rice, P(D54O), and tested the application of original and modified P(D54O) and
its cis-elements in the regulation of gene expression. P(D54O) is a green
tissue-specific promoter. Five novel tissue-specific cis-elements (LPSE1, LPSE2,
LPSRE1, LPSRE2, PSE1) were characterized from P(D54O). LPSE1 activated gene
expression in leaf and young panicle. LPSRE2 suppressed gene expression in leaf,
root, young panicle and stem, and PSE1 suppressed gene expression in young
panicle and stem. LPSRE1 and LPSE2 had dual roles in the regulation of
tissue-specific gene expression; both functioned as activators in leaf, but
LPSRE1 acted as a repressor in stem and LPSE2 as a repressor in young panicle and
root. Transgenic rice plants carrying cry1Ac encoding Bacillus thuringiensis
endotoxin, regulated by P(D54O), were resistant to leaf-folders, with no Cry1Ac
protein found in endosperm or embryo. A reporter gene regulated by a series of
truncated P(D54O) showed various tissue-specific expression patterns. Different
fragments of P(D54O) fused with the constitutive cauliflower mosaic virus 35S
promoter suppressed 35S-regulated gene expression in various tissues. P(D54O),
truncated P(D54O) and the tissue-specific cis-elements provide useful tools for
the regulation of tissue-specific gene expression in rice breeding programmes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17596180 [PubMed - indexed for MEDLINE]

483: Proc Natl Acad Sci U S A. 2007 Jul 3;104(27):11376-81. Epub 2007 Jun 26.

Conserved noncoding genomic sequences associated with a flowering-time
quantitative trait locus in maize.

Salvi S, Sponza G, Morgante M, Tomes D, Niu X, Fengler KA, Meeley R, Ananiev EV,
Svitashev S, Bruggemann E, Li B, Hainey CF, Radovic S, Zaina G, Rafalski JA,
Tingey SV, Miao GH, Phillips RL, Tuberosa R.

Department of Agroenvironmental Science and Technology, University of Bologna,
Viale Fanin, 44, 40127 Bologna, Italy. silvio.salvi@unibo.it

Flowering time is a fundamental trait of maize adaptation to different
agricultural environments. Although a large body of information is available on
the map position of quantitative trait loci for flowering time, little is known
about the molecular basis of quantitative trait loci. Through positional cloning
and association mapping, we resolved the major flowering-time quantitative trait
locus, Vegetative to generative transition 1 (Vgt1), to an approximately 2-kb
noncoding region positioned 70 kb upstream of an Ap2-like transcription factor
that we have shown to be involved in flowering-time control. Vgt1 functions as a
cis-acting regulatory element as indicated by the correlation of the Vgt1 alleles
with the transcript expression levels of the downstream gene. Additionally,
within Vgt1, we identified evolutionarily conserved noncoding sequences across
the maize-sorghum-rice lineages. Our results support the notion that changes in
distant cis-acting regulatory regions are a key component of plant genetic
adaptation throughout breeding and evolution.

Publication Types:
Comparative Study

PMID: 17595297 [PubMed - indexed for MEDLINE]

484: Mol Ecol. 2007 Jul;16(13):2617-26.

Comment in:
Mol Ecol. 2007 Jul;16(13):2607-9.

Estimating the number of whales entering trade using DNA profiling and
capture-recapture analysis of market products.

Baker CS, Cooke JG, Lavery S, Dalebout ML, Ma YU, Funahashi N, Carraher C,
Brownell RL Jr.

Marine Mammal Institute, Hatfield Marine Science Center, 2030 SE Marine Science
Drive, Oregon State University, Newport, OR 97365, USA.
scott.baker@oregonstate.edu

Surveys of commercial markets combined with molecular taxonomy (i.e. molecular
monitoring) provide a means to detect products from illegal, unregulated and/or
unreported (IUU) exploitation, including the sale of fisheries bycatch and wild
meat (bushmeat). Capture-recapture analyses of market products using DNA
profiling have the potential to estimate the total number of individuals entering
the market. However, these analyses are not directly analogous to those of living
individuals because a 'market individual' does not die suddenly but, instead,
remains available for a time in decreasing quantities, rather like the
exponential decay of a radioactive isotope. Here we use mitochondrial DNA (mtDNA)
sequences and microsatellite genotypes to individually identify products from
North Pacific minke whales (Balaenoptera acutorostrata ssp.) purchased in 12
surveys of markets in the Republic of (South) Korea from 1999 to 2003. By
applying a novel capture-recapture model with a decay rate parameter to the 205
unique DNA profiles found among 289 products, we estimated that the total number
of whales entering trade across the five-year survey period was 827 (SE, 164; CV,
0.20) and that the average 'half-life' of products from an individual whale on
the market was 1.82 months (SE, 0.24; CV, 0.13). Our estimate of whales in trade
(reflecting the true numbers killed) was significantly greater than the
officially reported bycatch of 458 whales for this period. This unregulated
exploitation has serious implications for the survival of this genetically
distinct coastal population. Although our capture-recapture model was developed
for specific application to the Korean whale-meat markets, the exponential decay
function could be modified to improve the estimates of trade in other wildmeat or
fisheries markets or abundance of living populations by noninvasive genotyping.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17594434 [PubMed - indexed for MEDLINE]

485: Plant Cell Rep. 2007 Oct;26(10):1745-53. Epub 2007 Jun 26.

Regeneration of transgenic plants from two indica rice (Oryza sativa L.)
cultivars using shoot apex explants.

Arockiasamy S, Ignacimuthu S.

Entomology Research Institute, Loyola College, Chennai 600034, India.

We have established a reproducible procedure for transformation of shoot apices
and regeneration of transgenic plants for two indica rice cultivars, white ponni
(WP) and Pusa Basmathi 1 (PB 1). Four-day-old shoot apex explants were
transformed by cocultivation with Agrobacterium tumefaciens strain EHA 101
harbouring a binary plasmid pRIT1. The vector contained an improved hygromycin
phosphotransferase (hpt) gene for hygromycin resistance driven by actin 1
promoter and the reporter gene beta-glucuronidase intron (INT-GUS) controlled by
CaMV 35S promoter. Rice shoots were induced on media containing 0.1 mg/l
napthalene acetic acid (NAA), 1.0 mg/l kinetin (kn), 1.0 mg/l
N(6)-benzyleaminopurin (BAP), 300 mg/l casaminoacid, 500 mg/l proline, 50 mg/l
hygromycin and 500 mg/l cefotaxime. Transgenic plants were raised in pots and
seeds were collected. Histochemical and polymerase chain reaction (PCR) analyses
of field established transgenic rice plants and their offsprings confirmed the
presence of GUS gene. Integration of T-DNA into the genome of putative
transgenics was further confirmed by southern analysis. The transformation
efficiency of WP was found to be ranging from 5.6 to 6.2% whereas in the case of
PB1, it was from 7 to 8%. Progeny analysis of these plants showed a pattern of
classical Mendelian inheritance for both hpt and GUS gene.

PMID: 17593368 [PubMed - indexed for MEDLINE]

486: Nat Biotechnol. 2007 Aug;25(8):899-901. Epub 2007 Jun 24.

Enrichment of tomato flavor by diversion of the early plastidial terpenoid
pathway.

Davidovich-Rikanati R, Sitrit Y, Tadmor Y, Iijima Y, Bilenko N, Bar E, Carmona B,
Fallik E, Dudai N, Simon JE, Pichersky E, Lewinsohn E.

Department of Vegetable Crops, Newe Ya'ar Research Center, Agricultural Research
Organization, PO Box 1021, Ramat Yishay, 30095 Israel.

We have modified the flavor and aroma of tomatoes by expressing the Ocimum
basilicum geraniol synthase gene under the control of the tomato
ripening-specific polygalacturonase promoter. A majority of untrained taste
panelists preferred the transgenic fruits over controls. Monoterpene accumulation
was at the expense of reduced lycopene accumulation. Similar approaches may be
applicable for carotenoid-accumulating fruits and flowers of other species.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17592476 [PubMed - indexed for MEDLINE]

487: J Agric Food Chem. 2007 Jul 25;55(15):6060-6. Epub 2007 Jun 23.

Evaluating homogeneity of LL601 rice in commercial lots using quantitative
real-time PCR.

Freese L, Scholdberg TA, Burton DD, Norden TD, Shokere LA, Jenkins GR.

Grain Inspection, Packers and Stockyards Administration, Technical Services
Division, U.S. Department of Agriculture, 10383 North Ambassador Drive, Kansas
City, Missouri 64153, USA.

Homogeneity analysis was performed on four distinctive commercial lots, derived
from the 2006 rice harvest in the United States. Lots that had previously been
tested and suspected to have some level of LL601 were selected to determine lot
homogeneity. LL601 infiltration in the lots was low and estimated to contain
<0.01% (sigma = 0.026), 0.014% (sigma = 0.020), 0.054% (sigma = 0.043), and
0.074% (sigma = 0.031) LL601. Lots were analyzed statistically as a one-way
classification, or one-factor experiment, to assess the presence of strata within
the lot. A p value of 0.05 or lower is needed to declare statistical significance
and would suggest significant differences among the samples. The data revealed p
values ranging between 0.105 and 0.607. The calculated p values for all lots were
greater than the critical value of 0.05. Samples taken from different locations
throughout these four commercial lots did not show statistically significant
stratifications within the lot.

PMID: 17590009 [PubMed - indexed for MEDLINE]

488: Mol Genet Genomics. 2007 Sep;278(3):243-54. Epub 2007 Jun 23.

ROSINA (RSI) is part of a CACTA transposable element, TamRSI, and links flower
development to transposon activity.

Roccaro M, Li Y, Sommer H, Saedler H.

Max-Planck-Institut fö¼r Zö¼chtungsforschung, Carl-von-Linne'-Weg 10, 50829 Koeln,
Germany. roccaro@mpiz-koeln.mpg.de

ROSINA (RSI) was isolated as a DNA binding factor able to bind to the CArG-box
present in the promoter of the MADS-box gene DEFICIENS of Antirrhinum majus. The
mosaic nature of RSI and its multi-copy presence in the A. majus genome indicated
that RSI could be a part of a mobile genetic element. Here we show that RSI is a
part of a CACTA transposable element system of A. majus, named TamRSI, which has
evolved and is still evolving within the terminal inverted repeats (TIRs) of this
CACTA transposon. Interestingly, RSI is always found in opposite orientation with
respect to the transcription of a second gene present within the CACTA
transposon, which encodes a putative TRANSPOSASE (TNP). This structural
configuration has not yet been described for any member of the CACTA transposons
superfamily. Internal deletion derivatives of the TamRSI produce aberrant RSI
transcripts (RSI-ATs) that carry parts of the RSI RNA fused to parts of the TNP
RNA. In addition, an intriguing seed phenotype shown by RNAi transgenic lines
generated to silence RSI, relate TamRSI to epigenetic mechanisms and associate
the control of flower development to transposon activity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17588178 [PubMed - indexed for MEDLINE]

489: Plant J. 2007 Aug;51(4):642-55. Epub 2007 Jun 22.

The cell cycle-associated protein kinase WEE1 regulates cell size in relation to
endoreduplication in developing tomato fruit.

Gonzalez N, Gö©vaudant F, Hernould M, Chevalier C, Mouras A.

Unitö© Mixte de Recherche 619 sur la Biologie du Fruit (Institut National de la
Recherche Agronomique; Universitö© Bordeaux 1; Universitö© Victor Segalen-Bordeaux
2), Institut Fö©dö©ratif de Recherche 103, Institut National de la Recherche
Agronomique, France.

Tomato fruit size results from the combination of cell number and cell size which
are respectively determined by cell division and cell expansion processes. As
fruit growth is mainly sustained by cell expansion, the development of pericarp
and locular tissues is characterized by the concomitant arrest of mitotic
activity, inhibition of cyclin-dependent kinase (CDK) activity, and numerous
rounds of endoreduplication inducing a spectacular increase in DNA ploidy and
mean cell size. To decipher the molecular basis of the
endoreduplication-associated cell growth in fruit, we investigated the putative
involvement of the WEE1 kinase (Solly;WEE1). We here report a functional analysis
of Solly;WEE1 in tomato. Impairing the expression of Solly;WEE1 in transgenic
tomato plants resulted in a reduction of plant size and fruit size. In the most
altered phenotypes, fruits displayed a reduced number of seeds without embryo
development. The reduction of plant-, fruit- and seed size originated from a
reduction in cell size which could be correlated with a decrease of the DNA
ploidy levels. At the molecular level downregulating Solly;WEE1 in planta
resulted in the increase of CDKA activity levels originating from a decrease of
the amount of Y15-phosphorylated CDKA, thus indicating a release of the negative
regulation on CDK activity exerted by WEE1. Our data indicated that Solly;WEE1
participates in the control of cell size and/or the onset of the
endoreduplication process putatively driving cell expansion.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17587306 [PubMed - indexed for MEDLINE]

490: Plant J. 2007 Aug;51(4):617-30. Epub 2007 Jun 22.

Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic
and biotic stress-responsive gene expression in rice.

Nakashima K, Tran LS, Van Nguyen D, Fujita M, Maruyama K, Todaka D, Ito Y,
Hayashi N, Shinozaki K, Yamaguchi-Shinozaki K.

Biological Resources Division, Japan International Research Center for
Agricultural Sciences, Tsukuba, Ibaraki 305-8686, Japan.

The OsNAC6 gene is a member of the NAC transcription factor gene family in rice.
Expression of OsNAC6 is induced by abiotic stresses, including cold, drought and
high salinity. OsNAC6 gene expression is also induced by wounding and blast
disease. A transactivation assay using a yeast system demonstrated that OsNAC6
functions as a transcriptional activator, and transient localization studies with
OsNAC6-sGFP fusion protein revealed its nuclear localization. Transgenic rice
plants over-expressing OsNAC6 constitutively exhibited growth retardation and low
reproductive yields. These transgenic rice plants showed an improved tolerance to
dehydration and high-salt stresses, and also exhibited increased tolerance to
blast disease. By utilizing stress-inducible promoters, such as the OsNAC6
promoter, it is hoped that stress-inducible over-expression of OsNAC6 in rice can
improve stress tolerance by suppressing the negative effects of OsNAC6 on growth
under normal growth conditions. The results of microarray analysis revealed that
many genes that are inducible by abiotic and biotic stresses were upregulated in
rice plants over-expressing OsNAC6. A transient transactivation assay showed that
OsNAC6 activates the expression of at least two genes, including a gene encoding
peroxidase. Collectively, these results indicate that OsNAC6 functions as a
transcriptional activator in response to abiotic and biotic stresses in plants.
We conclude that OsNAC6 may serve as a useful biotechnological tool for the
improvement of stress tolerance in various kinds of plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17587305 [PubMed - indexed for MEDLINE]

491: Plant J. 2007 Aug;51(4):670-80. Epub 2007 Jun 22.

Erratum in:
Plant J. 2008 Jan;53(2):400.

TaVRT2 represses transcription of the wheat vernalization gene TaVRN1.

Kane NA, Agharbaoui Z, Diallo AO, Adam H, Tominaga Y, Ouellet F, Sarhan F.

Dö©partement des Sciences Biologiques, Universitö© du Quö©bec ö  Montrö©al, Case
Postale 8888, Succursale Centre-ville, Montrö©al, QC H3C 3P8, Canada.

In wheat, VRN1/TaVRN1 and VRN2/TaVRN2 determine the growth habit and flowering
time. In addition, the MADS box transcription factor VEGETATIVE TO REPRODUCTIVE
TRANSITION 2 (TaVRT2) is also associated with the vernalization response in a
manner similar to TaVRN2. However, the molecular relationship between these three
genes and their products is unknown. Using transient expression assays in
Nicotiana benthamiana, we show that TaVRT2 acts as a repressor of TaVRN1
transcription. TaVRT2 binds the CArG motif in the TaVRN1 promoter and represses
its activity in vivo. In contrast, TaVRN2 does not bind the TaVRN1 promoter and
has no direct effect on its activity, but it can enhance the repression effect of
TaVRT2. This suggests that a repressor complex regulates the expression of
TaVRN1. In winter wheat, TaVRT2, TaVRN2 and TaVRN1 transcripts accumulate in the
shoot apical meristem and young leaves, and temporal expression is consistent
with TaVRT2 and TaVRN2 being repressors of floral transition, whereas TaVRN1 is
an activator. Non-vernalized spring wheat grown under a short-day photoperiod
accumulates TaVRT2 and shows a delay in flowering, suggesting that TaVRT2 is
regulated independently by photoperiod and low temperature. The data presented
suggest that TaVRT2, in association with TaVRN2, represses the transcription of
TaVRN1.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17587304 [PubMed - indexed for MEDLINE]

492: Plant J. 2007 Sep;51(5):803-18. Epub 2007 Jun 22.

Silencing of the major family of NBS-LRR-encoding genes in lettuce results in the
loss of multiple resistance specificities.

Wroblewski T, Piskurewicz U, Tomczak A, Ochoa O, Michelmore RW.

The Genome Center, University of California in Davis, Davis, CA 95616, USA.

The RGC2 gene cluster in lettuce (Lactuca sativa) is one of the largest known
families of genes encoding nucleotide binding site-leucine-rich repeat (NBS-LRR)
proteins. One of its members, RGC2B, encodes Dm3 which determines resistance to
downy mildew caused by the oomycete Bremia lactucae carrying the cognate
avirulence gene, Avr3. We developed an efficient strategy for analysis of this
large family of low expressed genes using post-transcriptional gene silencing
(PTGS). We transformed lettuce cv. Diana (carrying Dm3) using chimeric gene
constructs designed to simultaneously silence RGC2B and the GUS reporter gene via
the production of interfering hairpin RNA (ihpRNA). Transient assays of GUS
expression in leaves accurately predicted silencing of both genes and were
subsequently used to assay silencing in transgenic T(1) plants and their
offspring. Levels of mRNA were reduced not only for RGC2B but also for all seven
diverse RGC2 family members tested. We then used the same strategy to show that
the resistance specificity encoded by the genetically defined Dm18 locus in
lettuce cv. Mariska is the result of two resistance specificities, only one of
which was silenced by ihpRNA derived from RGC2B. Analysis of progeny from crosses
between transgenic, silenced tester stocks and lettuce accessions carrying other
resistance genes previously mapped to the RGC2 locus indicated that two
additional resistance specificities to B. lactucae, Dm14 and Dm16, as well as
resistance to lettuce root aphid (Pemphigus bursarius L.), Ra, are encoded by
RGC2 family members.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17587302 [PubMed - indexed for MEDLINE]

493: Plant J. 2007 Aug;51(3):468-84. Epub 2007 Jun 21.

Antisense inhibition of the plastidial glucose-6-phosphate/phosphate translocator
in Vicia seeds shifts cellular differentiation and promotes protein storage.

Rolletschek H, Nguyen TH, Hö¤usler RE, Rutten T, Gö¶bel C, Feussner I, Radchuk R,
Tewes A, Claus B, Klukas C, Linemann U, Weber H, Wobus U, Borisjuk L.

Leibniz-Institut fö¼r Pflanzengenetik und Kulturpflanzenforschung (IPK),
Corrensstr. 3, 06466 Gatersleben, Germany.

The glucose-6-phosphate/phosphate translocator (GPT) acts as an importer of
carbon into the plastid. Despite the potential importance of GPT for storage in
crop seeds, its regulatory role in biosynthetic pathways that are active during
seed development is poorly understood. We have isolated GPT1 from Vicia
narbonensis and studied its role in seed development using a transgenic approach
based on the seed-specific legumin promoter LeB4. GPT1 is highly expressed in
vegetative sink tissues, flowers and young seeds. In the embryo, localized
upregulation of GPT1 at the onset of storage coincides with the onset of starch
accumulation. Embryos of transgenic plants expressing antisense GPT1 showed a
significant reduction (up to 55%) in the specific transport rate of
glucose-6-phosphate as determined using proteoliposomes prepared from embryos.
Furthermore, amyloplasts developed later and were smaller in size, while the
expression of genes encoding plastid-specific translocators and proteins involved
in starch biosynthesis was decreased. Metabolite analysis and stable isotope
labelling demonstrated that starch biosynthesis was also reduced, although
storage protein biosynthesis increased. This metabolic shift was characterized by
upregulation of genes related to nitrogen uptake and protein storage,
morphological variation of the protein-storing vacuoles, and a crude protein
content of mature seeds of transgenics that was up to 30% higher than in
wild-type. These findings provide evidence that (1) the prevailing level of GPT1
abundance/activity is rate-limiting for the synthesis of starch in developing
seeds, (2) GPT1 exerts a controlling function on assimilate partitioning into
storage protein, and (3) GPT1 is essential for the differentiation of embryonic
plastids and seed maturation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17587237 [PubMed - indexed for MEDLINE]

494: Plant J. 2007 Aug;51(4):656-69. Epub 2007 Jun 21.

Heterotrimeric G-protein complex and G-protein-coupled receptor from a legume
(Pisum sativum): role in salinity and heat stress and cross-talk with
phospholipase C.

Misra S, Wu Y, Venkataraman G, Sopory SK, Tuteja N.

Plant Molecular Biology Group, International Centre for Genetic Engineering and
Biotechnology, New Delhi - 67, India.

Heterotrimeric G-proteins transduce signals from activated G-protein-coupled
receptors (GPCR) to appropriate downstream effectors and thereby play an
important role in signaling. A role of G-proteins in salinity and heat stress
tolerance has not heretofore been described. We report isolation of cDNAs of two
isoforms of Galpha (Galpha1, 1152 bp; Galpha2, 1152 bp), one Gbeta (1134 bp), two
isoforms of Ggamma (Ggamma1, 345 bp; Ggamma2, 303 bp) and a GPCR (1008 bp) from
Pisum sativum, and purification of all the encoded recombinant proteins (Galpha,
44 kDa; Gbeta, 41 kDa; Ggamma, 14 kDa; GPCR, 35 kDa). The transcript levels of
Galpha and Gbeta were upregulated following NaCl, heat and H(2)O(2) treatments.
Protein-protein interaction studies using an in vitro yeast two-hybrid system and
in planta co-immunoprecipitation showed that the Galpha subunit interacted with
the pea Gbeta subunit and pea phospholipase C (PLCdelta) at the calcium-binding
domain (fn1). The GTPase activity of the Galpha subunit increased after
interaction with PLCdelta. The GPCR protein interacted with all the subunits of
G-proteins and with itself. Transgenic tobacco plants (T(0) and T(1))
constitutively over-expressing Galpha showed tolerance to salinity and heat,
while Gbeta-over-expressing plants showed only heat tolerance, as tested by leaf
disk senescence assay and germination/growth of T(1) seeds/seedlings. These
findings provide direct evidence for a novel role of Galpha and Gbeta subunits in
abiotic stress tolerance and possible cross-talk between PLC- and
G-protein-mediated signaling pathways.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17587233 [PubMed - indexed for MEDLINE]

495: Plant J. 2007 Sep;51(5):792-802. Epub 2007 Jun 21.

Capsicum annuum CCR4-associated factor CaCAF1 is necessary for plant development
and defence response.

Sarowar S, Oh HW, Cho HS, Baek KH, Seong ES, Joung YH, Choi GJ, Lee S, Choi D.

Plant Genome Research Center, KRIBB, Daejeon, Korea.

The CCR4-associated factor 1 (CAF1) protein belongs to the CCR4-NOT complex,
which is an evolutionary conserved protein complex and plays an important role in
the control of transcription and mRNA decay in yeast and mammals. To investigate
the function of CAF1 in plants, we performed gain- and loss-of-function studies
by overexpression of the pepper CAF1 (CaCAF1) in tomato and virus-induced gene
silencing (VIGS) of the gene in pepper plants. Overexpression of CaCAF1 in tomato
resulted in significant growth enhancement, with increasing leaf thickness, and
enlarged cell size by more than twofold when compared with the control plants. A
transmission electron microscopic analysis revealed that the CaCAF1-transgenic
tomato plants had thicker cell walls and cuticle layers than the control plants.
In addition to developmental changes, overexpression of CaCAF1 in tomato plants
resulted in enhanced resistance against the oomycete pathogen Phytophthora
infestans. Additionally, microarray, northern and real-time polymerase chain
reaction analyses of CaCAF1-transgenic tomato plants revealed that multiple genes
were constitutively upregulated, including genes involved in polyamine
biosynthesis, defence reactions and cell-wall organogenesis. In contrast, VIGS of
CaCAF1 in pepper plants caused significant growth retardation and enhanced
susceptibility to the pepper bacterial spot pathogen Xanthomonas axonopodis pv.
vesicatoria. Our results suggest roles for plant CAF1 in normal growth and
development, as well as in defence against pathogens.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17587232 [PubMed - indexed for MEDLINE]

496: Plant Cell. 2007 Jun;19(6):1723-37. Epub 2007 Jun 22.

Genetic and epigenetic alteration among three homoeologous genes of a class E
MADS box gene in hexaploid wheat.

Shitsukawa N, Tahira C, Kassai K, Hirabayashi C, Shimizu T, Takumi S, Mochida K,
Kawaura K, Ogihara Y, Murai K.

Department of Bioscience, Fukui Prefectural University, Fukui, Japan.

Bread wheat (Triticum aestivum) is a hexaploid species with A, B, and D ancestral
genomes. Most bread wheat genes are present in the genome as triplicated
homoeologous genes (homoeologs) derived from the ancestral species. Here, we
report that both genetic and epigenetic alterations have occurred in the
homoeologs of a wheat class E MADS box gene. Two class E genes are identified in
wheat, wheat SEPALLATA (WSEP) and wheat LEAFY HULL STERILE1 (WLHS1), which are
homologs of Os MADS45 and Os MADS1 in rice (Oryza sativa), respectively. The
three wheat homoeologs of WSEP showed similar genomic structures and expression
profiles. By contrast, the three homoeologs of WLHS1 showed genetic and
epigenetic alterations. The A genome WLHS1 homoeolog (WLHS1-A) had a structural
alteration that contained a large novel sequence in place of the K domain
sequence. A yeast two-hybrid analysis and a transgenic experiment indicated that
the WLHS1-A protein had no apparent function. The B and D genome homoeologs,
WLHS1-B and WLHS1-D, respectively, had an intact MADS box gene structure, but
WLHS1-B was predominantly silenced by cytosine methylation. Consequently, of the
three WLHS1 homoeologs, only WLHS1-D functions in hexaploid wheat. This is a
situation where three homoeologs are differentially regulated by genetic and
epigenetic mechanisms.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17586655 [PubMed - indexed for MEDLINE]

497: Plant Cell. 2007 Jun;19(6):1770-81. Epub 2007 Jun 22.

Suppression of RICE TELOMERE BINDING PROTEIN 1 results in severe and gradual
developmental defects accompanied by genome instability in rice.

Hong JP, Byun MY, Koo DH, An K, Bang JW, Chung IK, An G, Kim WT.

Department of Biology, College of Science, Yonsei University, Seoul, Korea.

Although several potential telomere binding proteins have been identified in
higher plants, their in vivo functions are still unknown at the plant level. Both
knockout and antisense mutants of RICE TELOMERE BINDING PROTEIN1 (RTBP1)
exhibited markedly longer telomeres relative to those of the wild type,
indicating that the amount of functional RTBP1 is inversely correlated with
telomere length. rtbp1 plants displayed progressive and severe developmental
abnormalities in both germination and postgermination growth of vegetative organs
over four generations (G1 to G4). Reproductive organ formation, including
panicles, stamens, and spikelets, was also gradually and severely impaired in G1
to G4 mutants. Up to 11.4, 17.2, and 26.7% of anaphases in G2, G3, and G4 mutant
pollen mother cells, respectively, exhibited one or more chromosomal fusions, and
this progressively increasing aberrant morphology was correlated with an
increased frequency of anaphase bridges containing telomeric repeat DNA.
Furthermore, 35S:anti-RTBP1 plants expressing lower levels of RTBP1 mRNA
exhibited developmental phenotypes intermediate between the wild type and mutants
in all aspects examined, including telomere length, vegetative and reproductive
growth, and degree of genomic anomaly. These results suggest that RTBP1 plays
dual roles in rice (Oryza sativa), as both a negative regulator of telomere
length and one of positive and functional components for proper architecture of
telomeres.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17586654 [PubMed - indexed for MEDLINE]

498: Physiol Behav. 2007 Nov 23;92(4):691-701. Epub 2007 May 21.

Lentivirus-mediated downregulation of hypothalamic insulin receptor expression.

Grillo CA, Tamashiro KL, Piroli GG, Melhorn S, Gass JT, Newsom RJ, Reznikov LR,
Smith A, Wilson SP, Sakai RR, Reagan LP.

Department of Pharmacology, Physiology and Neuroscience, University of South
Carolina, Columbia, SC, United States.

Regulation of feeding behavior and energy balance are among the central effects
of insulin. For example, intracerebroventricular administration of insulin
decreases food intake and body weight, whereas antisense oligodeoxynucleotide
downregulation of insulin receptors (IRs) produces hyperphagia. To further
examine the role of IRs in the central actions of insulin, we designed an IR
antisense lentiviral vector (LV-IRAS) and injected this vector into the third
ventricle to selectively decrease IR expression in the rat hypothalamus. Three
weeks after LV-IRAS administration, the expression of IRs in the hypothalamus was
significantly decreased, whereas no changes were observed in hippocampal IR
levels. LV-IRAS administration decreased insulin-stimulated phosphorylation of
hypothalamic IRs and translocation of the insulin-sensitive glucose transporter
GLUT4 in the hypothalamus; no changes in IR signaling were observed in the
hippocampus of LV-IRAS-treated rats. Lentivirus-mediated downregulation of IR
expression and signaling produced significant increases in body weight, as well
as increases in fat mass that were selective for the subcutaneous compartment.
Conversely, lean muscle mass and water mass were not affected in LV-IRAS-treated
rats compared to rats treated with control virus. Changes in peripheral adiposity
were associated with increases in basal hypothalamic leptin signaling in the
absence of changes in leptin receptor expression in LV-IRAS rats. Collectively,
these data illustrate the important functional relationships between hypothalamic
insulin and leptin signaling in the regulation of body composition and provide
insight into the mechanisms through which decreases in IR expression and
signaling dysregulates leptin activity, thereby promoting increases in peripheral
adiposity.

Publication Types:
Evaluation Studies
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17585961 [PubMed - indexed for MEDLINE]

499: Indian J Exp Biol. 2007 Jun;45(6):554-62.

Effect of Bt-cotton on chrysopids, ladybird beetles and their prey: aphids and
whiteflies.

Mellet MA, Schoeman AS.

Department of Zoology & Entomology, University of Pretoria, Pretoria, 0002, South
Africa. magdel.mellet@bayercropscience.com

The effect of Bt-cotton, i.e. genetically modified cotton that contain genes
expressing delta-endotoxin, on aphid, whitefly, chrysopid and coccinellid
populations was determined with a two-year field study at a cotton farm near
Marble Hall, South Africa. Although Bt-cotton is lepidopteran specific,
non-lepidopteran arthropod populations may be indirectly influenced by the
endotoxin. Abundance of aphid, whitefly, chrysopid and coccinellid populations
and predator-prey interactions were used as measures to determine possible
effects on the populations under investigation. The cultivation of Bt-cotton had
no effect on aphid, whitefly, chrysopid or coccinellid abundance. Positive
density dependent interactions occurred between aphids and coccinellids which
were not influenced by Bt-cotton. A significant relationship between whitefly and
coccinellid abundance, i.e. predator-prey reaction, occurred in the control and
sprayed non-Bt cotton fields but was absent from the Bt-cotton fields.

PMID: 17585692 [PubMed - indexed for MEDLINE]

500: Nat Protoc. 2007;2(7):1614-21.

Agrobacterium-mediated transformation of maize.

Ishida Y, Hiei Y, Komari T.

Plant Innovation Center, Japan Tobacco Inc., 700 Higashibara, Iwata, Shizuoka,
Japan. yuji.a.ishida@ims.jti.co.jp

Maize may be transformed very efficiently using Agrobacterium
tumefaciens-mediated methods. The most critical factor in the transformation
protocol is the co-cultivation of healthy immature embryos of the correct
developmental stage with A. tumefaciens; the embryos should be collected only
from vigorous plants grown in well-conditioned glasshouses. With the protocol
described here, approximately 50% of immature embryos from the inbred line A188
and 15% from inbred lines A634, H99 and W117 will produce transformants. About
half of the transformed plants are expected to carry one or two copies of the
transgenes, which are inherited by the progeny in a mendelian fashion. More than
90% of transformants are expected to be normal in morphology. The protocol takes
about 3 months from the start of co-cultivation to the planting of transformants
into pots.

PMID: 17585302 [PubMed - indexed for MEDLINE]

501: J Agric Food Chem. 2007 Jul 25;55(15):6074-81. Epub 2007 Jun 20.

Suppression of C-hordein synthesis in barley by antisense constructs results in a
more balanced amino acid composition.

Lange M, Vincze E, Wieser H, Schjoerring JK, Holm PB.

Faculty of Agricultural Sciences, Department of Genetics and Biotechnology,
Research Centre Flakkebjerg, University of Aarhus, Forsoegsvej 1, DK-4200
Slagelse, Denmark. mette.lange@agrsci.dk

Barley has for feeding purposes a shortage of essential amino acids, especially
lysine, threonine, and methionine, and an excess of proline and glutamine. In the
present study, we have introduced into barley an antisense construct against
C-hordeins, the storage protein with the lowest nutritional quality. SDS-PAGE and
reverse phase HPLC revealed a relative reduction in the amounts of C-hordeins and
relative increases in the content of the other storage proteins. The five
different lines analyzed had lower amounts of proline, glutamic acid/glutamine,
and phenylalanine (up to 12%, 6%, and 9% reductions), while the lysine,
threonine, and methionine content was increased with up to 16%, 13% and 11%. It
is concluded that antisense mediated suppression of C-hordein synthesis may be a
promising approach for improving the nutritional value of barley as a feed crop
while at the same time reducing the environmental nitrogen load.

PMID: 17580876 [PubMed - indexed for MEDLINE]

502: J AOAC Int. 2007 May-Jun;90(3):794-801.

Real-time polymerase chain reaction assay for endogenous reference gene for
specific detection and quantification of common wheat-derived DNA (Triticum
aestivum L.).

Vautrin S, Zhang D.

GEVES, Domaine du Magneraud, Laboratoire BioGEVES, B.P. 52, F-17700 Surgeres,
France.

A species-specific endogenous reference gene system was developed for polymerase
chain reaction (PCR)-based analysis in common wheat (Triticum aestivum L.) by
targeting the ALMT1 gene, an aluminium-activated malate transporter. The primers
and probe were elaborated for real-time PCR-based qualitative and quantitative
assay. The size of amplified product is 95 base pairs. The specificity was
assessed on 17 monocot and dicot plant species. The established real-time PCR
assay amplified only T. aestivum-derived DNA; no amplification occurred on other
phylogenetically related species, including durum wheat (T. durum). The
robustness of the system was tested on the DNA of 15 common wheat cultivars using
20 000 genomic copies per PCR the mean cycle threshold (Ct) values of 24.02 +/-
0.251 were obtained. The absolute limits of detection and quantification of the
real-time PCR assay were estimated to 2 and 20 haploid genome copies of common
wheat, respectively. The linearity was experimentally validated on 2-fold serial
dilutions of DNA from 650 to 20 000 haploid genome copies. All these results show
that the real-time PCR assay developed on the ALMT1 gene is suitable to be used
as an endogenous reference gene for PCR-based specific detection and
quantification of T. aestivum-derived DNA in various applications, in particular
for the detection and quantification of genetically modified materials in common
wheat.

PMID: 17580632 [PubMed - indexed for MEDLINE]

503: Proc Natl Acad Sci U S A. 2007 Jun 26;104(26):10962-7. Epub 2007 Jun 19.

Transposition of the rice miniature inverted repeat transposable element mPing in
Arabidopsis thaliana.

Yang G, Zhang F, Hancock CN, Wessler SR.

Department of Plant Biology, University of Georgia, Athens, GA 30602, USA.

An active miniature inverted repeat transposable element (MITE), mPing, was
discovered by computer-assisted analysis of rice genome sequence. The mPing
element is mobile in rice cell culture and in a few rice strains where it has
been amplified to >1,000 copies during recent domestication. However,
determination of the transposase source and characterization of the mechanism of
transposition have been hampered by the high copy number of mPing and the
presence of several candidate autonomous elements in the rice genome. Here, we
report that mPing is active in Arabidopsis thaliana, where its transposition is
catalyzed by three sources of transposase from rice: the autonomous Ping and Pong
elements and by a cDNA derived from a Ping transcript. In addition to
transposase, the product of a second element-encoded ORF of unknown function is
also required for mPing transposition. Excision of mPing in A. thaliana is
usually precise, and transposed copies usually insert into unlinked sites in the
genome that are preferentially in or near genes. As such, this will be a valuable
assay system for the dissection of MITE transposition and a potentially powerful
tagging system for gene discovery in eukaryotes.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17578919 [PubMed - indexed for MEDLINE]

504: Food Chem Toxicol. 2007 Oct;45(10):1994-2004. Epub 2007 May 6.

Evaluation of the safety and nutritional equivalence of a genetically modified
cottonseed meal in a 90-day dietary toxicity study in rats.

Dryzga MD, Yano BL, Andrus AK, Mattsson JL.

Toxicology and Environmental Research and Consulting, The Dow Chemical Company,
Midland, MI 48674, USA. MDDryzga@dow.com

Meal prepared from Cry1F/Cry1Ac transgenic/genetically modified cottonseed
(WIDESTRIKE Insect Protection, hereafter referred to as WIDESTRIKE) was compared
to cottonseed meal prepared from four conventionally bred lines of cotton (three
commercial non-transgenic line controls (PHY72, PHY78 and 98M-2983), and a near
isoline non-transgenic control (PSC355) in a 90-day dietary study to evaluate
safety and nutritional equivalence. Diets were formulated with 10% WIDESTRIKE
cottonseed meal equivalent to 7,235 mg/kg/day for males and 7,935 mg/kg/day for
females. Animals were evaluated by cage-side and hand-held detailed clinical
observations, body weight, and feed consumption. Functional tests, motor activity
and ophthalmic examinations were conducted pre-exposure and prior to study
termination. Standard hematology, clinical chemistry, prothrombin time and
urinalysis parameters were evaluated. All rats had a complete necropsy and
selected organs were weighed. Histopathologic examinations were performed on all
rats fed the diets containing the near isoline non-transgenic control or
WIDESTRIKE. Following 90 days of feeding, no adverse effects were observed during
the conduct of clinical observations or in any of the parameters measured in this
study. This study demonstrated that rodent diets prepared with 10% cottonseed
meal from WIDESTRIKE cottonseeds do not produce any untoward effects and are
nutritionally equivalent to cottonseed meals prepared from other, non-transgenic
cottonseeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17574718 [PubMed - indexed for MEDLINE]

505: Theriogenology. 2007 Sep 1;68 Suppl 1:S3-8. Epub 2007 Jun 15.

Regulation of animal biotechnology: research needs.

Rexroad CE Jr, Green RD, Wall RJ.

U.S. Department of Agriculture, Agricultural Research Service, 5601 Sunnyside
Avenue, Rm. 4-2150, Beltsville, MD 20705-5134, USA. caird.rexroad@ars.usda.gov

Livestock that result from biotechnology have been a part of agricultural science
for over 30 years but have not entered the market place as food or fiber. Two
biotechnologies are at the forefront as challenges to the world's systems for
regulating the market place: animal clones and transgenic animals. Both
technologies have come before the Food and Drug Administration in the United
States and it appears that action is imminent for clones. The FDA has asserted
principles for evaluation of clones and asserts that "... remaining hazard(s)
from cloning are likely to be subtle in nature." The science-based principles
recognize that in some areas related to developmental biology and gene expression
in clones, additional scientific information would be useful. The role of science
then is to use the genomic tools that we have available to answer questions about
epigenetic regulation of development and reprogramming of genes to the state
found in germ cells. Transgenics pose additional challenges to regulators. If the
transgenics are produced using cloning from modified cells then the additional
scientific information needed will be related to the effects of insertion and
expression of the transgenes. Other approaches such as retrovirally vectored
transgenesis will elicit additional questions. These questions will be
challenging because the science will have to be related to the expression and
function of each gene or class of genes. For the promises of animal biotechnology
to be fulfilled, scientists will have to resolve many questions for regulators
and the public but tools to answer those questions are rapidly becoming
available.

Publication Types:
Review

PMID: 17574657 [PubMed - indexed for MEDLINE]

506: Mol Genet Genomics. 2007 Oct;278(4):411-20. Epub 2007 Jun 16.

Molecular analysis of Agrobacterium T-DNA integration in tomato reveals a role
for left border sequence homology in most integration events.

Thomas CM, Jones JD.

School of Biological Sciences, University of East Anglia, Norwich, NR4 7TJ, UK.
colwyn.thomas@uea.ac.uk

Studies in several plants have shown that Agrobacterium tumefaciens T-DNA can
integrate into plant chromosomal DNA by different mechanisms involving
single-stranded (ss) or double-stranded (ds) forms. One mechanism requires
sequence homology between plant target and ssT-DNA border sequences and another
double-strand-break repair in which preexisting chromosomal DSBs "capture"
dsT-DNAs. To learn more about T-DNA integration in Solanum lycopersicum we
characterised 98 T-DNA/plant DNA junction sequences and show that T-DNA left
border (LB) and right border transfer is much more variable than previously
reported in Arabidopsis thaliana and Populus tremula. The analysis of seven plant
target sequences showed that regions of homology between the T-DNA LB and plant
chromosomal DNA plays an important role in T-DNA integration. One T-DNA insertion
generated a target sequence duplication that resulted from nucleolytic processing
of a LB/plant DNA heteroduplex that generated a DSB in plant chromosomal DNA. One
broken end contained a captured T-DNA that served as a template for DNA repair
synthesis. We propose that most T-DNA integrations in tomato require sequence
homology between the ssT-DNA LB and plant target DNA which results in the
generation of DSBs in plant chromosomal DNA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17574477 [PubMed - indexed for MEDLINE]

507: Nat Biotechnol. 2007 Jul;25(7):759-61. Epub 2007 Jun 17.

Comment in:
Nat Biotechnol. 2007 Jul;25(7):746-8.

Lignin modification improves fermentable sugar yields for biofuel production.

Chen F, Dixon RA.

Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway,
Ardmore, Oklahoma 73401, USA. fchen@noble.org

Recalcitrance to saccharification is a major limitation for conversion of
lignocellulosic biomass to ethanol. In stems of transgenic alfalfa lines
independently downregulated in each of six lignin biosynthetic enzymes,
recalcitrance to both acid pretreatment and enzymatic digestion is directly
proportional to lignin content. Some transgenics yield nearly twice as much sugar
from cell walls as wild-type plants. Lignin modification could bypass the need
for acid pretreatment and thereby facilitate bioprocess consolidation.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17572667 [PubMed - indexed for MEDLINE]

508: Nutr Cancer. 2007;58(1):66-74.

Transgenic alfalfa that accumulates piceid
(trans-resveratrol-3-O-beta-D-glucopyranoside) requires the presence of
beta-glucosidase to inhibit the formation of aberrant crypt foci in the colon of
CF-1 mice.

Kineman BD, Au A, Paiva NL, Kaiser MS, Brummer EC, Birt DF.

Department of Food Science and Human Nutrition, Iowa State University, Ames, IA
50011, USA.

Plants have been genetically enhanced to produce a number of products for
agricultural, industrial and pharmaceutical purposes. This technology could
potentially be applied to providing chemoprevention strategies to the general
population. Resveratrol (3,5,4'-trihydroxystilbene) is a compound that has been
shown to have protective activity against a number of cancers and could be an
ideal candidate for such an application. Alfalfa that was genetically modified to
express resveratrol-synthase was used as a model in applying biotechnological
approaches to cancer prevention. The transgenic alfalfa, which accumulates
resveratrol as a glucoside (piceid =
trans-resveratrol-3-O-Beta-D-glucopyranoside) (152 +/- 17.5 microg piceid/g dry
weight), was incorporated into a standard mouse diet at 20% of the diet by weight
and fed for 5 wk to 6-wk-old, female CF-1 mice (N = 17-30) that were injected
with a single dose of azoxymethane (5 mg/kg body weight). While the addition of
resveratrol-aglycone (20 mg/kg diet) to the basal diet reduced the number of
aberrant crypt foci/mouse, the transgenic alfalfa did not inhibit the number,
size, or multiplicity of aberrant crypt foci in the colon of the CF-1 mice
relative to control alfalfa which does not accumulate resveratrol-glucoside.
However, diets containing transgenic alfalfa with an exogenous Beta-glucosidase
(860 U/kg diet) did significantly inhibit the number of aberrant crypt foci in
the distal 2 cm of the colon of the mice relative to mice fed diets containing
the transgenic alfalfa without the enzyme (P < 0.05; Fisher's Combination of
p-values). The Beta-glucosidase alone appeared to have no effect on the
inhibition of aberrant crypt foci. These results suggest that piceid in
transgenic piceid-accumulating alfalfa was not bioavailable.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17571969 [PubMed - indexed for MEDLINE]

509: Plant Cell Rep. 2007 Oct;26(10):1909-17. Epub 2007 Jun 15.

Two rice cytosolic ascorbate peroxidases differentially improve salt tolerance in
transgenic Arabidopsis.

Lu Z, Liu D, Liu S.

Biochemistry and Molecular Biology Lab, College of Life Sciences, Heilongjiang
University, Harbin 150080, People's Republic China.

In order to determine the different roles of rice (Oryza sativa L.) cytosolic
ascorbate peroxidases (OsAPXa and OsAPXb, GenBank accession nos. D45423 and
AB053297, respectively) under salt stress, transgenic Arabidopsis plants
over-expressing OsAPXa or OsAPXb were generated, and they all exhibited increased
tolerance to salt stress compared to wild-type plants. Moreover, transgenic lines
over-expressing OsAPXb showed higher salt tolerance than OsAPXa transgenic lines
as indicated by root length and total chlorophyll content. In addition to
ascorbate peroxidase (APX) activity, antioxidant enzyme activities of catalase
(CAT), superoxide dismutase (SOD) and glutathione reductase (GR), which are also
involved in the salt tolerance process, and the content of H2O2 were also assayed
in both transgenic and wild-type plants. The results showed that the
overproduction of OsAPXb enhanced and maintained APX activity to a much higher
degree than OsAPXa in transgenic Arabidopsis during treatment with different
concentrations of NaCl, enhanced the active oxygen scavenging system, and
protected plants from salt stress by equilibrating H2O2 metabolism. Our findings
suggest that the rice cytosolic OsAPXb gene has a more functional role than
OsAPXa in the improvement of salt tolerance in transgenic plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17571267 [PubMed - indexed for MEDLINE]

510: Biotechnol J. 2007 Jun;2(6):670.

Opinion: EU biotech regulations stall agricultural improvements in developing
countries.

[No authors listed]

Publication Types:
News

PMID: 17570700 [PubMed - indexed for MEDLINE]

511: Biotechnol J. 2007 Jun;2(6):653.

Engineering plant oils.

[No authors listed]

Publication Types:
News

PMID: 17570698 [PubMed - indexed for MEDLINE]

512: J Plant Physiol. 2008 Feb;165(2):159-71. Epub 2007 Jun 13.

Induction of a polyubiquitin gene promoter by dehydration stresses in transformed
rice cells.

Perales L, Peö±arrubia L, Cornejo MJ.

Departmento de Biologö­a Vegetal, Facultad de Biologö­a, Avda. Dr Moliner 50, 46100
Burjasot, Valencia, Spain.

The expression of the maize polyubiquitin gene promoter UBI1 in rice cells has
been used to study the involvement of ubiquitin in cell protection responses to
dehydration caused by osmotic, saline or freezing stress. The effect of these
stresses on UBI1 activity was investigated by the use of stably transformed rice
calli (UBI1:GUS), as well as by transient expression experiments performed with
cell lines with high or low tolerance to each type of stress. The theoretical
analysis of the UBI1 promoter shows several putative stress-regulated boxes that
could account for the stress-related UBI1 induction pattern described in this
work. We suggest that the study of the differential UBI1 promoter-driven
expression in rice cell lines with different level of tolerance to stress might
be useful to elucidate complex signal transduction pathways in response to
dehydration stresses in monocots.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17570562 [PubMed - indexed for MEDLINE]

513: Plant Cell Rep. 2007 Oct;26(10):1733-44. Epub 2007 Jun 14.

Stable chloroplast transformation in cabbage (Brassica oleracea L. var. capitata
L.) by particle bombardment.

Liu CW, Lin CC, Chen JJ, Tseng MJ.

Department of Post-Modern Agriculture, Ming Dao University, Chang Hua 523,
Taiwan, ROC. chad888@mud.edu.tw

The objectives of this research were first to isolate plastid gene sequences from
cabbage (Brassica oleracea L. var. capitata L.), and to establish the chloroplast
transformation technology of Brassica. A universal transformation vector
(pASCC201) for Brassica chloroplast was constructed with trnV-rrn16S (left) and
trnI-trnA-rrn23S (right) of the IR(_A) region as a recombination site for the
transformed gene. In transforming plasmid pASCC201, a chimeric aadA gene was
cloned between the rrn16S and rrn23S plastid gene borders. Expression of aadA
confers resistance to spectinomycin and streptomycin antibiotics. The uidA gene
was also inserted into the pASCC201 and transferred into the leaf cells of
cabbage via particle gun mediated transformation. Regenerated plantlets were
selected by 200 mg/l spectinomycin and streptomycin. After antibiotic selection,
the regeneration percentage of the two cabbage cultivars was about 2.7-3.3%. The
results of PCR testing and Southern blot analysis confirmed that the uidA and
aadA genes were present in the chloroplast genome via homologously recombined.
Northern blot hybridizations, immunoblotting and GUS histochemical assays
indicated that the uidA gene were stable integrated into the chloroplast genome.
Foreign protein was accumulated at 3.2-5.2% of the total soluble protein in
transgenic mature leaves. These results suggest that the expression of a variety
of foreign genes in the chloroplast genome will be a powerful tool for use in
future studies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17569052 [PubMed - indexed for MEDLINE]

514: Plant Cell Rep. 2007 Oct;26(10):1763-71. Epub 2007 Jun 14.

Expression of a feedback insensitive anthranilate synthase gene from tobacco
increases free tryptophan in soybean plants.

Inaba Y, Brotherton JE, Ulanov A, Widholm JM.

Faculty of Medicine, Dentistry and Health Sciences, Department of Pediatrics,
University of Melbourne, Melbourne, Australia.

Soybean [Glycine max (L.) Merr.] embryogenic cultures were transformed by
particle bombardment with the feedback-insensitive tobacco anthranilate synthase
(AS) gene ASA2 driven by the CaMV 35S promoter and selected using hph as the
selectable marker gene. Only one of eight regenerated lines that set seed and
contained ASA2 expressed the gene highly and contained increased free tryptophan
(Trp) levels in leaves, seeds and embryogenic cultures. Leaf extracts of the ASA2
expressing line contained about twice as much AS enzyme activity as the
untransformed control and this activity was only slightly more
feedback-insensitive. Amino acid analysis showed that both leaves and embryogenic
tissue cultures of the ASA2 expressing line had four to five-times the normal
levels of free Trp and slightly higher free tyrosine and phenylalanine. The seed
total Trp content was only slightly increased. Metabolic profiling-analysis by
GC-MS detected no other consistent differences. These studies show that the ASA2
gene can be expressed in soybean and that modest changes in Trp synthesis occurs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17569051 [PubMed - indexed for MEDLINE]

515: Plant Cell Rep. 2007 Oct;26(10):1773-83. Epub 2007 Jun 14.

Heterologous expression of Vitreoscilla haemoglobin in barley (Hordeum vulgare).

Wilhelmson A, Kallio PT, Oksman-Caldentey KM, Nuutila AM.

VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT, Espoo,
Finland. annika.wilhelmson@vtt.fi

The vhb gene encoding Vitreoscilla haemoglobin (VHb) was transferred to barley
with the aim of studying the role of oxygen availability in germination and
growth. Previous findings indicate that VHb expression improves the efficiency of
energy generation during oxygen-limited growth, and germination is known to be an
energy demanding growth stage during which the embryos also suffer from oxygen
deficiency. When subjected to oxygen deficiency, the roots of vhb-expressing
barley plants showed a smaller increase in alcohol dehydrogenase (ADH) activity
than those of the control plants. This indicates that VHb plants experienced less
severe oxygen deficiency than the control plants, possibly due to the ability of
VHb to substitute ADH for recycling NADH and maintaining glycolysis. In contrast
to previous findings, we found that constitutive vhb expression did not improve
the germination rate of barley kernels in any of the conditions studied. In some
cases, vhb expression even slowed down germination slightly. VHb production also
appeared to restrict root formation in young seedlings. The adverse effects of
VHb on germination and root growth may be related to its ability to scavenge
nitric oxide (NO), an important signal molecule in both seed germination and root
formation. Because NO has both cytotoxic and stimulating properties, the effect
of vhb expression in plants may depend on the level and role of endogenous NO in
the conditions studied. VHb production also affected the levels of endogenous
barley haemoglobin, which may explain the relatively moderate effects of VHb in
this study.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17569049 [PubMed - indexed for MEDLINE]

516: Appl Microbiol Biotechnol. 2007 Sep;76(3):607-13. Epub 2007 Jun 14.

Purification of recombinant aprotinin from transgenic corn germ fraction using
ion exchange and hydrophobic interaction chromatography.

Zhong Q, Xu L, Zhang C, Glatz CE.

Department of Chemical and Biological Engineering, Iowa State University, Ames,
IA 50011, USA. qzhong@utk.edu

Plants have attracted interest as hosts for protein expression because of the
promise of a large production capacity and a low production cost. However,
recovery costs remain a challenge as illustrated for recovery of recombinant
aprotinin, a trypsin inhibitor, with removal of native corn trypsin inhibitor
from transgenic corn (Azzoni et al. in Biotechnol Bioeng 80:268-276, 2002). When
expression is targeted to corn grain fractions, dry milling can separate germ and
endosperm fractions. Hence, only the product-containing fraction needs to be
extracted, reducing the cost of extraction and the impurity level of the extract.
Selective extraction conditions can reduce impurity levels to the point that
low-cost adsorbents can result in relatively high purity levels. In this work, we
attempted to achieve comparable purity with these lower cost methods. We replaced
whole grain extraction and purification of recombinant aprotinin with sequential
trypsin affinity and IMAC steps with an alternative of germ fraction extraction
and purification with ion exchange and hydrophobic interaction chromatography
(HIC). Using germ extraction at acidic pH supplemented with heat precipitation to
remove additional host proteins resulted in a higher specific activity feed to
the chromatographic steps. The cation exchange step provided 7.6x purification
with 76.4% yield and no sodium dodecyl sulfate-polyacrylamide gel electrophoresis
detectable native corn trypsin inhibitor. After the HIC step (2.7x step
purification with 44.0% yield), the final product had a specific activity that
was 75.3% of that of the affinity-purified aprotinin.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17569041 [PubMed - indexed for MEDLINE]

517: Plant Cell Physiol. 2007 Jul;48(7):1050-60. Epub 2007 Jun 13.

Characterization of silencing suppressor 2b of cucumber mosaic virus based on
examination of its small RNA-binding abilities.

Goto K, Kobori T, Kosaka Y, Natsuaki T, Masuta C.

Cell Biology and Manipulation Laboratory, Graduate School of Agriculture,
Hokkaido University, Sapporo, 060-8589 Japan.

Double-stranded (ds) RNAs and imperfect hairpin RNAs of endogenous genes trigger
post-transcriptional gene silencing (PTGS) and are cleaved by a Dicer-like
nuclease into small interfering RNAs (siRNAs) and microRNs (miRNAs),
respectively. Such small RNAs (siRNAs and miRNAs) then guide an RNA-induced
silencing complex (RISC) for sequence-specific RNA degradation. While PTGS serves
as an antiviral defense in plants, many plant viruses encode suppressors as a
counter defense. Here we demonstrate that the PTGS suppressor (2b) of a severe
strain (CM95R) of cucumber mosaic virus (CMV) can bind to in vitro synthesized
siRNAs and even to long dsRNAs to a lesser extent. However, the 2b suppressor
weakly bound to a miRNA (miR171) duplex in contrast to another small RNA-binding
suppressor, p19 of tombusvirus that can effectively bind miRNAs. Because the 2b
suppressor of an attenuated strain of CMV (CM95), which differs in a single amino
acid from the 2b of CM95R, could barely bind siRNAs, we hypothesized that the
weak suppressor activity of the attenuated strain resulted from a loss of the
siRNA-binding property of 2b via a single amino acid change. Here we consider
that 2b interferes with the PTGS pathway by directly binding siRNAs (or long
dsRNA).

PMID: 17567638 [PubMed - indexed for MEDLINE]

518: J Plant Physiol. 2007 Oct;164(10):1384-90. Epub 2007 Jun 12.

Over-expression of OsUGE-1 altered raffinose level and tolerance to abiotic
stress but not morphology in Arabidopsis.

Liu HL, Dai XY, Xu YY, Chong K.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, The Chinese Academy of Sciences, Nanxincun 20, Xiangshan,
Beijing 100093, China.

OsUGE-1 is known to be induced by various abiotic stresses, but its exact
function in plants is unclear. In the present study, OsUGE-1 was over-expressed
in Arabidopsis, transgenic plants conferred tolerance to salt, drought and
freezing stress without altering plant morphology. In addition, transgenic plants
showed a higher level of the soluble sugar raffinose than did wild-type plants.
Our results suggest that elevated level of raffinose with over-expressed OsUGE-1
resulted in enhanced tolerance to abiotic stress. Thus, the gene may be applied
to improve tolerance to abiotic stress in crops.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17566602 [PubMed - indexed for MEDLINE]

519: J Biotechnol. 2007 Jun 30;130(3):236-46. Epub 2007 Apr 24.

Regulation of alcohol oxidase of a recombinant Pichia pastoris Mut+ strain in
transient continuous cultures.

Jungo C, Marison I, von Stockar U.

Ecole Polytechnique Fö©dö©rale de Lausanne, Laboratoire de Gö©nie Chimique et
Biologique, Station 6, CH-1015 Lausanne, Switzerland.

In the methylotrophic yeast Pichia pastoris, alcohol oxidase (AOX) is a key
enzyme involved in the dissimilation of methanol. Heterologous proteins are
usually expressed under the control of the AOX1 promoter, which drives the
expression of alcohol oxidase 1 in the wild-type strain. This study investigates
the regulation of the alcohol oxidase enzyme of a recombinant P. pastoris Mut+
strain in cultures on glycerol and methanol as sole carbon sources and in mixed
substrate cultures on both substrates. The aim was to have a better insight in
the transition from growth on glycerol to growth on methanol, which is a key step
in standard high cell density P. pastoris cultures for the production of foreign
proteins. Nutrient shifts in chemostat cultures showed that after growth on
glycerol use of mixed feeds of glycerol and methanol allowed faster induction of
alcohol oxidase and faster adaptation of cellular metabolism than with a feed
containing methanol as sole carbon source. The results of this study showed also
how critical it is to avoid transient methanol accumulation during P. pastoris
cultures operated at low residual methanol concentrations. Indeed, pulse
experiments during chemostat cultures showed that sudden increase in methanol
concentrations in cultures performed under methanol-limited or dual methanol and
glycerol-limited growth conditions leads to wash-out of the culture because of
too high consumption rate of methanol, which leads to excretion of toxic
intermediates. High rate of methanol consumption was due to high specific AOX
activities observed at low residual methanol concentrations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17566583 [PubMed - indexed for MEDLINE]

520: Biochim Biophys Acta. 2007 Jul-Aug;1769(7-8):455-61. Epub 2007 May 21.

A conserved role of SHORT VEGETATIVE PHASE (SVP) in controlling flowering time of
Brassica plants.

Lee JH, Park SH, Lee JS, Ahn JH.

Plant Signaling Network Research Center, School of Life Sciences and
Biotechnology, Korea University, Seoul 136-701, South Korea.

The control of flowering time in Brassica plants is an important approach for
improving productivity, as vegetative tissues are not produced after the floral
transition in Brassica plants. In order to determine the feasibility of
modulating flowering time in Chinese cabbage plants, genes homologous to
Arabidopsis SHORT VEGETATIVE PHASE (AtSVP) were isolated from spring-type and
fall-type cultivars of Chinese cabbage plants, and their functions were
determined. Their deduced amino acid sequences were 91-93% identical with that of
AtSVP. The expression of BcSVP was ubiquitously detected, and was unaffected by
vernalization. Constitutive BcSVP expression induced late flowering with
additional floral defects. This delayed flowering was attributed to the
repression of FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS
1 (SOC1). BcSVP expression under the control of the AtSVP promoter also resulted
in the complementation of the svp mutation in Arabidopsis. These results indicate
that BcSVP is a functional equivalent of AtSVP and also suggest that BcSVP may
prove useful for the genetic manipulation of flowering time in Brassica plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17566572 [PubMed - indexed for MEDLINE]

521: Br J Soc Psychol. 2007 Jun;46(Pt 2):437-57.

Predicting behaviour towards genetically modified food using implicit and
explicit attitudes.

Spence A, Townsend E.

RASPH, School of Psychology, University of Nottingham, UK. spenceAl@cardiff.ac.uk

The predictive validity of implicit and explicit attitudes is a central question
in social psychological research with important theoretical and empirical
ramifications. Three main patterns of combining implicit and explicit attitudes
to predict behaviour have been postulated. They are, double dissociation (in
which implicit and explicit attitudes predict spontaneous and deliberate
behaviour respectively), additive (in which implicit and explicit attitudes both
predict variance in behaviour) and interactive (in which implicit and explicit
attitudes combine to predict behaviour). These models were tested in this study
using a structural equation modelling approach utilising three different measures
of behaviour (of varying spontaneity) towards genetically modified (GM) food. The
additive pattern, in which implicit and explicit attitudes both predict variance
in behaviour, was found to best fit the data. In addition, all behaviour measures
indicated that the majority of participants were willing to try GM food in some
situations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17565791 [PubMed - indexed for MEDLINE]

522: Plant Biotechnol J. 2007 Sep;5(5):636-45. Epub 2007 Jun 12.

Cloning and functional characterization of the fatty acid elongase 1 (FAE1) gene
from high erucic Crambe abyssinica cv. Prophet.

Mietkiewska E, Brost JM, Giblin EM, Barton DL, Taylor DC.

Department of Plant Sciences, University of Saskatchewan, 51 Campus Drive,
Saskatoon, Saskatchewan, Canada S7N 5A8.

A genomic fatty acid elongation 1 (FAE1) clone was isolated from Crambe
abyssinica. The genomic clone corresponds to a 1521-bp open reading frame, which
encodes a protein of 507 amino acids. In yeast cells expression of CrFAE led to
production of new very long chain monounsaturated fatty acids such as eicosenoic
(20:1(delta11)) and erucic (22:1(delta13)) acids. Seed-specific expression in
Arabidopsis thaliana resulted in up to a 12-fold increase in the proportion of
erucic acid. On the other hand, in transgenic high-erucic Brassica carinata
plants, the proportion of erucic acid was as high as 51.9% in the best transgenic
line, a net increase of 40% compared to wild type. These results indicate that
the CrFAE gene encodes a condensing enzyme involved in the biosynthesis of very
long-chain fatty acids utilizing monounsaturated and saturated acyl substrates,
with a strong capability for improving the erucic acid content.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17565584 [PubMed - indexed for MEDLINE]

523: J Exp Bot. 2007;58(10):2699-707. Epub 2007 Jun 11.

C-terminal extension of rice glutamate decarboxylase (OsGAD2) functions as an
autoinhibitory domain and overexpression of a truncated mutant results in the
accumulation of extremely high levels of GABA in plant cells.

Akama K, Takaiwa F.

Department of Biological Science, Shimane University, Nishikawatsu 1060, Matsue,
Shimane, Japan. akama@life.shimane-u.ac.jp

Glutamate decarboxylase (GAD) converts L-glutamate to gamma-aminobutyric acid
(GABA), which is a non-protein amino acid present in all organisms. Plant GADs
carry a C-terminal extension that binds to Ca(2+)/calmodulin (CaM) to modulate
enzyme activity. However, rice possesses two distinct types of GAD, OsGAD1 and
OsGAD2. Although they both have a C-terminal extension, the former peptide
contains an authentic CaM-binding domain (CaMBD), which is common to
dicotyledonous plants, while the latter does not. Therefore, the role of the
C-terminal extension in functional expression of OsGAD2 was investigated. An in
vitro enzyme assay using recombinant OsGAD2 proteins revealed low activity in the
presence or absence of Ca(2+)/CaM. However, a truncated version of GAD2
(OsGAD2DeltaC) had over 40-fold higher activity than wild-type GAD at
physiological pH. These two DNA constructs were introduced simultaneously into
rice calli via Agrobacterium to establish transgenic cell lines. Free amino acids
were isolated from several lines for each construct to determine GABA content.
Calli overexpressing OsGAD2 and OsGAD2DeltaC had about 6-fold and 100-fold the
GABA content of wild-type calli, respectively. Regenerated OsGAD2DeltaC rice
plants had aberrant phenotypes such as dwarfism, etiolated leaves, and sterility.
These data suggest that the C-terminal extension of OsGAD2 plays a role as a
strong autoinhibitory domain, and that truncation of this domain causes the
enzyme to act constitutively, with higher activity both in vitro and in vivo.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17562689 [PubMed - indexed for MEDLINE]

524: J Exp Bot. 2007;58(10):2573-82. Epub 2007 Jun 11.

Isolation and comparative analysis of the wheat TaPT2 promoter: identification in
silico of new putative regulatory motifs conserved between monocots and dicots.

Tittarelli A, Milla L, Vargas F, Morales A, Neupert C, Meisel LA, Salvo-G H,
Peö±aloza E, Muö±oz G, Corcuera LJ, Silva H.

Millennium Nucleus in Plant Cell Biology and Center of Plant Biotechnology,
Andres Bello University, Av Repöºblica 217, 837-0146, Santiago, Chile.

Phosphorus deficiency is one of the major nutrient stresses affecting plant
growth. Plants respond to phosphate (Pi) deficiency through multiple strategies,
including the synthesis of high-affinity Pi transporters. In this study, the
expression pattern of one putative wheat high-affinity phosphate transporter,
TaPT2, was examined in roots and leaves under Pi-deficient conditions. TaPT2
transcript levels increased in roots of Pi-starved plants. A 579 bp fragment of
the TaPT2 promoter is sufficient to drive the expression of the GUS reporter gene
specifically in roots of Pi-deprived wheat. This TaPT2 promoter fragment was also
able to drive expression of the GUS reporter gene in transgenic Arabidopsis
thaliana, under similar growth conditions. Conserved regions and candidate
regulatory motifs were detected by comparing this promoter with Pi transporter
promoters from barley, rice, and Arabidopsis. Altogether, these results indicate
that there are conserved cis-acting elements and trans-acting factors that enable
the TaPT2 promoter to be regulated in a tissue-specific and Pi-dependent fashion
in both monocots and dicots.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17562688 [PubMed - indexed for MEDLINE]

525: J Environ Sci Health B. 2007 Jun-Jul;42(5):539-49.

Review of potential environmental impacts of transgenic glyphosate-resistant
soybean in Brazil.

Cerdeira AL, Gazziero DL, Duke SO, Matallo MB, Spadotto CA.

Brazilian Department of Agriculture, Agricultural Research Service, Jaguariöºna,
SP, Brazil. cerdeira@cnpma.embrapa.br

Transgenic glyphosate-resistant soybeans (GRS) have been commercialized and grown
extensively in the Western Hemisphere, including Brazil. Worldwide, several
studies have shown that previous and potential effects of glyphosate on
contamination of soil, water, and air are minimal, compared to those caused by
the herbicides that they replace when GRS are adopted. In the USA and Argentina,
the advent of glyphosate-resistant soybeans resulted in a significant shift to
reduced- and no-tillage practices, thereby significantly reducing environmental
degradation by agriculture. Similar shifts in tillage practiced with GRS might be
expected in Brazil. Transgenes encoding glyphosate resistance in soybeans are
highly unlikely to be a risk to wild plant species in Brazil. Soybean is almost
completely self-pollinated and is a non-native species in Brazil, without wild
relatives, making introgression of transgenes from GRS virtually impossible.
Probably the highest agricultural risk in adopting GRS in Brazil is related to
weed resistance. Weed species in GRS fields have shifted in Brazil to those that
can more successfully withstand glyphosate or to those that avoid the time of its
application. These include Chamaesyce hirta (erva-de-Santa-Luzia), Commelina
benghalensis (trapoeraba), Spermacoce latifolia (erva-quente), Richardia
brasiliensis (poaia-branca), and Ipomoea spp. (corda-de-viola). Four weed
species, Conyza bonariensis, Conyza Canadensis (buva), Lolium multiflorum
(azevem), and Euphorbia heterophylla (amendoim bravo), have evolved resistance to
glyphosate in GRS in Brazil and have great potential to become problems.

Publication Types:
Review

PMID: 17562462 [PubMed - indexed for MEDLINE]

526: Plant Biotechnol J. 2007 Sep;5(5):570-8. Epub 2007 Jun 11.

Oral immunization with transgenic rice seeds expressing VP2 protein of infectious
bursal disease virus induces protective immune responses in chickens.

Wu J, Yu L, Li L, Hu J, Zhou J, Zhou X.

State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang
University, Hangzhou 31009, China.

The expression of infectious bursal disease virus (IBDV) host-protective
immunogen VP2 protein in rice seeds, its immunogenicity and protective capability
in chickens were investigated. The VP2 cDNA of IBDV strain ZJ2000 was cloned
downstream of the Gt1 promoter of the rice glutelin GluA-2 gene in the binary
expression vector, pCambia1301-Gt1. Agrobacterium tumefaciens containing the
recombinant vector was used to transform rice embryogenic calli, and 121
transgenic lines were obtained and grown to maturity in a greenhouse. The
expression level of VP2 protein in transgenic rice seeds varied from 0.678% to
4.521% microg/mg of the total soluble seed protein. Specific pathogen-free
chickens orally vaccinated with transgenic rice seeds expressing VP2 protein
produced neutralizing antibodies against IBDV and were protected when challenged
with a highly virulent IBDV strain, BC6/85. These results demonstrate that
transgenic rice seeds expressing IBDV VP2 can be used as an effective, safe and
inexpensive vaccine against IBDV.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17561926 [PubMed - indexed for MEDLINE]

527: J Biotechnol. 2007 Oct 31;132(2):180-6. Epub 2007 Apr 29.

Process Analytical Technology (PAT): batch-to-batch reproducibility of
fermentation processes by robust process operational design and control.

Gnoth S, Jenzsch M, Simutis R, Lö¼bbert A.

Center for Bioprocess Engineering, Martin-Luther-University Halle-Wittenberg,
Germany.

The Process Analytical Technology (PAT) initiative of the FDA is a reaction on
the increasing discrepancy between current possibilities in process supervision
and control of pharmaceutical production processes and its current application in
industrial manufacturing processes. With rigid approval practices based on
standard operational procedures, adaptations of production reactors towards the
state of the art were more or less inhibited for long years. Now PAT paves the
way for continuous process and product improvements through improved process
supervision based on knowledge-based data analysis, "Quality-by-Design"-concepts,
and, finally, through feedback control. Examples of up-to-date implementations of
this concept are presented. They are taken from one key group of processes in
recombinant pharmaceutical protein manufacturing, the cultivations of genetically
modified Escherichia coli bacteria.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17559961 [PubMed - indexed for MEDLINE]

528: Plant Biotechnol J. 2007 Sep;5(5):591-604. Epub 2007 Jun 8.

Erratum in:
Plant Biotechnol J. 2007 Sep;5(5):676.

Use of a stress inducible promoter to drive ectopic AtCBF expression improves
potato freezing tolerance while minimizing negative effects on tuber yield.

Pino MT, Skinner JS, Park EJ, Jekniä‡ Z, Hayes PM, Thomashow MF, Chen TH.

Department of Horticulture, ALS 4017, Oregon State University, Corvallis, OR
97331, USA.

Solanum tuberosum is a frost-sensitive species incapable of cold acclimation. A
brief exposure to frost can significantly reduce its yields, while hard frosts
can completely destroy entire crops. Thus, gains in freezing tolerance of even a
few degrees would be of considerable benefit relative to frost damage. The S.
tuberosum cv. Umatilla was transformed with three Arabidopsis CBF genes
(AtCBF1-3) driven by either a constitutive CaMV35S or a stress-inducible
Arabidopsis rd29A promoter. AtCBF1 and AtCBF3 over-expression via the 35S
promoter increased freezing tolerance about 2 degrees C, whereas AtCBF2
over-expression failed to increase freezing tolerance. Transgenic plants of
AtCBF1 and AtCBF3 driven by the rd29A promoter reached the same level of freezing
tolerance as the 35S versions within a few hours of exposure to low but
non-freezing temperatures. Constitutive expression of AtCBF genes was associated
with negative phenotypes, including smaller leaves, stunted plants, delayed
flowering, and reduction or lack of tuber production. While imparting the same
degree of freezing tolerance, control of AtCBF expression via the
stress-inducible promoter ameliorated these negative phenotypic effects and
restored tuber production to levels similar to wild-type plants. These results
suggest that use of a stress-inducible promoter to direct CBF transgene
expression can yield significant gains in freezing tolerance without negatively
impacting agronomically important traits in potato.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17559519 [PubMed - indexed for MEDLINE]

529: Plant J. 2007 Jul;51(2):198-210. Epub 2007 Jun 8.

A role for the AtMTP11 gene of Arabidopsis in manganese transport and tolerance.

Delhaize E, Gruber BD, Pittman JK, White RG, Leung H, Miao Y, Jiang L, Ryan PR,
Richardson AE.

CSIRO Plant Industry, Canberra, ACT, Australia. manny.delhaize@csiro.au

The Arabidopsis AtMTP family of genes encode proteins of the cation diffusion
facilitator (CDF) family, with several members having roles in metal tolerances.
Four of the 11 proteins in the family form a distinct cluster on a phylogenetic
tree and are closely related to ShMTP8, a CDF identified in the tropical legume
Stylosanthes hamata that is implicated in the transport of Mn(2+) into the
vacuole as a tolerance mechanism. Of these four genes, AtMTP11 was the most
highly expressed member of the Arabidopsis subgroup. When AtMTP11 was expressed
in Saccharomyces cerevisiae, it conferred Mn(2+) tolerance and transported Mn(2+)
by a proton-antiport mechanism. A mutant of Arabidopsis with a disrupted AtMTP11
gene (mtp11) was found to have increased sensitivity to Mn(2+) but not to Cu(2+)
or Zn(2+). At a non-toxic but sufficient Mn(2+) supply (basal), the mutant
accumulated more Mn(2+) than the wild type, but did not show any obvious
deleterious effects on growth. When grown with Mn(2+) supplies that ranged from
basal to toxic, the mutant accumulated Mn(2+) concentrations in shoots similar to
those in wild-type plants, despite showing symptoms of Mn(2+) toxicity. AtMTP11
fused to green fluorescent protein co-localized with a reporter specific for
pre-vacuolar compartments. These findings provide evidence for Mn(2+)-specific
transport activity by AtMTP11, and implicate the pre-vacuolar compartments in
both Mn(2+) tolerance and Mn(2+) homeostasis mechanisms of Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17559518 [PubMed - indexed for MEDLINE]

530: Plant J. 2007 Aug;51(3):366-77. Epub 2007 Jun 8.

The rice bHLH protein OsIRO2 is an essential regulator of the genes involved in
Fe uptake under Fe-deficient conditions.

Ogo Y, Itai RN, Nakanishi H, Kobayashi T, Takahashi M, Mori S, Nishizawa NK.

Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1
Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

Iron (Fe) deficiency is a major abiotic stress in crop production. Although
responses to Fe deficiency in graminaceous plants, such as increased production
and secretion of mugineic acid family phytosiderophores (MAs), have been
described, the gene regulation mechanisms related to these responses are largely
unknown. To elucidate the regulation mechanisms of the genes related to Fe
acquisition in graminaceous plants, we characterized the Fe-deficiency-inducible
basic helix-loop-helix transcription factor OsIRO2 in rice. In yeast cells,
OsIRO2 functioned as a transcriptional activator. In rice, overexpression of
OsIRO2 resulted in increased MAs secretion, whereas repression of OsIRO2 resulted
in lower MAs secretion and hypersensitivity to Fe deficiency. Northern blots
revealed that the expression of the genes involved in the Fe(III)-MAs transport
system was dependent on OsIRO2. The expression of the genes for nicotianamine
synthase, a key enzyme in MAs synthesis, was notably affected by the level of
OsIRO2 expression. Microarray analysis demonstrated that OsIRO2 regulates 59
Fe-deficiency-induced genes in roots. Some of these genes, including two
transcription factors upregulated by Fe deficiency, possessed the OsIRO2 binding
sequence in their upstream regions. OsIRO2 possesses a homologous sequence of the
Fe-deficiency-responsive cis-acting elements (IDEs) in its upstream region. We
propose a novel gene regulation network for Fe-deficiency responses, including
OsIRO2, IDEs and the two transcription factors.

PMID: 17559517 [PubMed - indexed for MEDLINE]

531: Plant J. 2007 Jul;51(1):92-104. Epub 2007 Jun 8.

RING-H2 type ubiquitin ligase EL5 is involved in root development through the
maintenance of cell viability in rice.

Koiwai H, Tagiri A, Katoh S, Katoh E, Ichikawa H, Minami E, Nishizawa Y.

Division of Plant Sciences, National Institute of Agrobiological Sciences,
Kannondai, 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.

Rice EL5 is an ATL family gene characterized by a transmembrane domain at the
N-terminal and a RING-H2 finger domain (RFD), which exhibits ubiquitin ligase
(E3) activity. To elucidate the physiological roles of EL5, we analyzed
transgenic rice plants overexpressing mutant EL5 proteins that are impaired in E3
activity to various degrees. Plants expressing EL5C153A and EL5W165A, which
encode an inactive E3, showed a rootless phenotype accompanied by cell death in
root primordia, and those expressing EL5V162A, with moderately impaired E3
activity, formed short crown roots with necrotic lateral roots. The
dominant-negative phenotype was specifically observed in root meristems where EL5
is expressed, and not recovered by exogenous auxin. When wild-type EL5 was
transcriptionally overexpressed, the EL5 protein was barely detected by Western
blotting. Neither treatment with a proteasome inhibitor nor mutation of the sole
lysine residue, a potential target of ubiquitination, resulted in increased EL5
accumulation, whereas mutations in the RFD led to increased EL5 accumulation. The
stabilized EL5 without the RFD was localized in the plasma membrane. Deletion of
the transmembrane domain prevented the EL5 from localizing in the membrane and
from exerting an inhibitory effect on root formation. Deletion of the C-terminal
region also neutralized the negative effect. We concluded that EL5 plays a major
role as a membrane-anchored E3 for the maintenance of cell viability after the
initiation of root primordial formation. In addition, we propose that EL5 is an
unstable protein, of which degradation is regulated by the RFD in a
proteasome-independent manner.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17559513 [PubMed - indexed for MEDLINE]

532: J Agric Food Chem. 2007 Jul 11;55(14):5575-9. Epub 2007 Jun 9.

Event-specific detection of seven genetically modified soybean and maizes using
multiplex-PCR coupled with oligonucleotide microarray.

Xu J, Zhu S, Miao H, Huang W, Qiu M, Huang Y, Fu X, Li Y.

State Key Laboratory of Genetic Engineering, Institute of Genetics, School of
Life Science, Fudan University, Shanghai 200433, P.R. China.

With the increasing development of genetically modified organism (GMO) detection
techniques, the polymerase chain reaction (PCR) technique has been the mainstay
for GMO detection. An oligonucleotide microarray is a glass chip to the surface
of which an array of oligonucleotides was fixed as spots, each containing
numerous copies of a sequence-specific probe that is complementary to a gene of
interest. So it is used to detect ten or more targets synchronously. In this
research, an event-specific detection strategy based on the unique and specific
integration junction sequences between the host plant genome DNA and the
integrated gene is being developed for its high specificity using multiplex-PCR
together with oligonucleotide microarray. A commercial GM soybean (GTS 40-3-2)
and six GM maize events (MON810, MON863, Bt176, Bt11, GA21, and T25) were
detected by this method. The results indicate that it is a suitable method for
the identification of these GM soybean and maizes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17559227 [PubMed - indexed for MEDLINE]

533: Crit Rev Food Sci Nutr. 2007;47(5):441-98.

Implementation of physicochemical and sensory analysis in conjunction with
multivariate analysis towards assessing olive oil authentication/adulteration.

Arvanitoyannis IS, Vlachos A.

University of Thessaly, School of Agricultural Sciences, Department of
Agriculture Animal Production and Aquatic Environment, Volos, Hellas, Greece.
parmenion@uth.gr

The authenticity of products labeled as olive oils, and in particular as virgin
olive oils, stands for a very important issue both in terms of its health and
commercial aspects. In view of the continuously increasing interest in virgin
olive oil therapeutic properties, the traditional methods of characterization and
physical and sensory analysis were further enriched with more advanced and
sophisticated methods such as HPLC-MS, HPLC-GC/C/IRMS, RPLC-GC, DEPT, and CSIA
among others. The results of both traditional and "novel" methods were treated
both by means of classical multivariate analysis (cluster, principal component,
correspondence, canonical, and discriminant) and artificial intelligence methods
showing that nowadays the adulteration of virgin olive oil with seed oil is
detectable at very low percentages, sometimes even at less than 1%. Furthermore,
the detection of geographical origin of olive oil is equally feasible and much
more accurate in countries like Italy and Spain where databases of
physical/chemical properties exist. However, this geographical origin
classification can also be accomplished in the absence of such databases provided
that an adequate number of oil samples are used and the parameters studied have
"discriminating power."

Publication Types:
Review

PMID: 17558656 [PubMed - indexed for MEDLINE]

534: Nat Biotechnol. 2007 Jun;25(6):624-6.

Comment on:
Nat Biotechnol. 2006 May;24(5):498; author reply 499.

Trends in GM crop, food and feed safety literature.

Vain P.

Publication Types:
Comment
Letter

PMID: 17557092 [PubMed - indexed for MEDLINE]

535: Nat Biotechnol. 2007 Jun;25(6):623; author reply 624.

Comment on:
Nat Biotechnol. 2006 Oct;24(10):1200-1.

What we know and don't know about Golden Rice.

Krawinkel MB.

Publication Types:
Comment
Letter

PMID: 17557091 [PubMed - indexed for MEDLINE]

536: Science. 2007 Jun 8;316(5830):1475-7.

A meta-analysis of effects of Bt cotton and maize on nontarget invertebrates.

Marvier M, McCreedy C, Regetz J, Kareiva P.

Environmental Studies Institute, Santa Clara University, Santa Clara, CA 95053,
USA. mmarvier@scu.edu

Although scores of experiments have examined the ecological consequences of
transgenic Bacillus thuringiensis (Bt) crops, debates continue regarding the
nontarget impacts of this technology. Quantitative reviews of existing studies
are crucial for better gauging risks and improving future risk assessments. To
encourage evidence-based risk analyses, we constructed a searchable database for
nontarget effects of Bt crops. A meta-analysis of 42 field experiments indicates
that nontarget invertebrates are generally more abundant in Bt cotton and Bt
maize fields than in nontransgenic fields managed with insecticides. However, in
comparison with insecticide-free control fields, certain nontarget taxa are less
abundant in Bt fields.

Publication Types:
Meta-Analysis
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17556584 [PubMed - indexed for MEDLINE]

537: Comp Biochem Physiol A Mol Integr Physiol. 2007 Sep;148(1):214-22. Epub 2007
Apr 14.

Comparison of metabolic rates and feed nutrient digestibility in conventional,
genetically improved (GIFT) and genetically male (GMNT) Nile tilapia, Oreochromis
niloticus (L.).

Mamun SM, Focken U, Becker K.

Department of Aquaculture Systems and Animal Nutrition, Institute for Animal
Production in the Tropics and Subtropics, University of Hohenheim (480B), D-70593
Stuttgart, Germany.

Various aspects of energy metabolism and feed digestibility were evaluated in two
reportedly improved strains of Nile tilapia (Oreochromis niloticus) namely GIFT
(genetically improved farmed tilapia) and GMNT (genetically male Nile tilapia)
and compared with those of CNT (conventional Nile tilapia). Fish were stocked
individually in a computer-controlled respirometer system at 27+/-0.1 degrees C
for 10 weeks. Metabolic rates were measured at three different feeding levels:
starved, maintenance (3.0 g kg(-0.8) day(-1)) and growth (7.5 g kg(-0.8) day(-1))
using a fishmeal based feed containing TiO2 marker (41% crude protein, 9% crude
lipid and 19 kJ (g DM)(-1) gross energy). The standard metabolic rate (SMR),
measured at the beginning of the experiment (45.4+/-4.6, 52.4+/-7.7 and
46.8+/-4.6 mg O2 kg(-0.8) h(-1) respectively for GIFT, GMNT and CNT), did not
differ significantly between the groups (p<0.05). Similarly, non-significant
differences were also observed in the routine metabolic rates under starved,
maintenance and growth conditions but the variability was higher in the case of
GMNT and CNT than in GIFT. The latter group showed a significantly lower active
metabolic rate (145 mg O2 kg(-0.8) h(-1)) compared to GMNT and CNT (232 and 253
mg O2 kg(-0.8) h(-1), respectively) at maintenance feeding level. The specific
dynamic action (% offered feed energy) showed no significant differences among
the groups. Digestibility coefficients of feed dry matter, protein, lipid and
energy for the three tilapia groups also did not differ significantly. Therefore,
we concluded that the genetic improvement or modification in the GIFT or GMNT
might not upgrade the inherent physiological potential compared to CNT as far as
energy metabolism and digestion efficiencies are concerned.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17555997 [PubMed - indexed for MEDLINE]

538: Ecol Appl. 2007 Jun;17(4):1234-43.

The effect of wind direction on cross-pollination in wind-pollinated GM crops.

Hoyle M, Cresswell JE.

School of Biosciences, University of Exeter, Exeter EX4 4PS, UK.
m.w.hoyle@exeter.ac.uk

In Europe, regulatory thresholds restrict adventitious GM (genetically modified)
presence in conventional crops. Minimum distances for the spatial separation of
fields are often recommended to reduce field-to-field cross-pollination to an
acceptable level. Field trials are typically the basis for setting separation
distances. However, using records of wind direction and speed from weather
stations across Europe, we predict theoretically that field-to-field windborne
cross-pollination in maize, oilseed rape, sugar beet, and rice varies greatly
according to the relative orientation of the GM and non-GM fields. Furthermore,
at a given site and orientation from a GM field, we predict that the
cross-pollination rate varies substantially from year to year. Consequently, even
replicated field trials may inaccurately estimate typical levels of
cross-pollination and therefore distort our perception of the separation
distances required to achieve sub-threshold adventitious GM presence. We propose
methods to predict the likely range in levels of cross-pollination based on the
limited data typically available from field trials. Additionally, we suggest
suitable time lags between peak flowering in adjacent fields that could be
introduced to reduce cross-pollination to a specified level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17555231 [PubMed - indexed for MEDLINE]

539: Plant Cell Rep. 2007 Oct;26(10):1839-59. Epub 2007 Jun 7.

Transgenic tobacco plants overexpressing the heterologous lea gene Rab16A from
rice during high salt and water deficit display enhanced tolerance to salinity
stress.

RoyChoudhury A, Roy C, Sengupta DN.

Department of Botany, Bose Institute, 93/1, Acharya Prafulla Chandra Road,
Kolkata 700009, West Bengal, India.

The full length Rab16A, from the indica rice Pokkali, was introduced into tobacco
by Agrobacterium-mediated transformation. The transgene was stably integrated
into the genome and they originated from different lines of integration.
Expression of Rab16A transcript driven by its own promoter (stress inducible) in
T2 progenies, only when triggered by salinity/ABA/PEG (Polyethylene
glycol)-mediated dehydration, but not at the constitutive level, led to the
stress-induced accumulation of RAB16A protein in the leaves of transgenic plants.
The selected independent transgenic lines showed normal growth, morphology and
seed production as the WT plants without any yield penalty under stress
conditions. They exhibited significantly increased tolerance to salinity,
sustained growth rates under stress conditions; with concomitant increased
osmolyte production like reducing sugars, proline and higher polyamines. They
also showed delayed development of damage symptoms with better antioxidative
machinery and more favorable mineral balance, as reflected by reduced H2O2 levels
and lipid peroxidation, lesser chlorophyll loss as well as lesser accumulation of
Na+ and greater accumulation of K+ in 200 mM NaCl. These findings establish the
potential role of Rab16A gene in conferring salt tolerance without affecting
growth and yield, as well as pointing to the fact that the upstream region of
Rab16A behaves as an efficient stress-inducible promoter. Our result also
suggests the considerable potential of Group 2 lea genes as molecular tools for
genetic engineering of plants towards stress tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17554543 [PubMed - indexed for MEDLINE]

540: J Gen Virol. 2007 Jul;88(Pt 7):2073-7.

Genomic regions of tomato leaf curl virus DNA satellite required for replication
and for satellite-mediated delivery of heterologous DNAs.

Li D, Behjatnia SA, Dry IB, Randles JW, Eini O, Rezaian MA.

CSIRO Plant Industry, PO Box 350, Glen Osmond, SA 5064, Australia.

Tomato leaf curl virus (TLCV) satellite DNA (sat-DNA) is a 682 nt, circular,
single-stranded molecule that lacks an open reading frame (ORF) or an apparent
promoter. It contains binding motifs for the TLCV replication-associated protein,
but these are dispensable for replication. To identify the regions of the sat-DNA
critical for replication, the entire sequence was scanned by deletion/replacement
mutagenesis. Transient assays using Nicotiana benthamiana revealed that sequences
within nt 296-35 (through nt 682) are essential for replication. Sequence
deletions and replacements between nt 35 and 296 were tolerated but with a
significant loss of infectivity, indicating that genome size strongly influences
replication efficiency. Within the permissible region, inserts of 100-700 nt were
retained in transient assays although with a slight reduction in replication. In
addition, sat-DNA constructs containing short non-viral DNAs replicated and
spread in tobacco plants, indicating their potential as gene-delivery vectors.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17554042 [PubMed - indexed for MEDLINE]

541: Med Hypotheses. 2007;69(6):1257-60. Epub 2007 Jun 5.

Are xenogeneic anti-tissue transglutaminase antibodies the holy grail for celiac
patients?

Ivanovski PI, Ivanovski IP, Sedlarevic R.

University Children's Hospital, 10 Tirshova Str., Belgrade, Serbia.
ivanovsk@eunet.yu

Celiac disease is an immune mediated disorder, the only one with a
well-established origin, resulting from a permanent gluten intolerance. Although
a gluten-free diet is currently the "safe" and appropriate therapy for celiac
disease, this is not always an easy and simple option as "harmful" gluten may
contaminate food during the processing and preparation phases. There are also
further social pressures, which might be more pressing for young celiac patients,
in following a strict gluten-free diet. Therefore, a new therapeutic approaches
are sought which would permit celiacs to "peacefully" coexist with gluten.
Presently, the most promising looks search for genetically modified wheat lacking
toxic gluten peptides and the use of oral endopeptidases in attempt to curb
gluten toxicity. Recently discovered role of anti-tissue transglutaminase
antibodies in celiac pathogenesis has brought a prospect for a new hypothetical
therapeutic approach, an oral immunization of celiacs with xenogeneic anti-tissue
transglutaminase antibodies.

PMID: 17553630 [PubMed - indexed for MEDLINE]

542: Plant Biotechnol J. 2007 Sep;5(5):605-14. Epub 2007 Jun 6.

High-lysine corn generated by endosperm-specific suppression of lysine catabolism
using RNAi.

Houmard NM, Mainville JL, Bonin CP, Huang S, Luethy MH, Malvar TM.

Mystic Research, Monsanto Company, 62 Maritime Drive, Mystic, CT 06355, USA.

Because of the limited lysine content in corn grain, synthetic lysine supplements
are added to corn meal-based rations for animal feed. The development of
biotechnology, combined with the understanding of plant lysine metabolism,
provides an alternative solution for increasing corn lysine content through
genetic engineering. Here, we report that by suppressing lysine catabolism,
transgenic maize kernels accumulated a significant amount of lysine. This was
achieved by RNA interference (RNAi) through the endosperm-specific expression of
an inverted-repeat (IR) sequence targeting the maize bifunctional lysine
degradation enzyme, lysine-ketoglutarate reductase/saccharopine dehydrogenase
(ZLKR/SDH). Although plant-short interfering RNA (siRNA) were reported to lack
tissue specificity due to systemic spreading, we confirmed that the suppression
of ZLKR/SDH in developing transgenic kernels was restricted to endosperm tissue.
Furthermore, results from our cloning and sequencing of siRNA suggested the
absence of transitive RNAi. These results support the practical use of RNAi for
plant genetic engineering to specifically target gene suppression in desired
tissues without eliciting systemic spreading and the transitive nature of plant
RNAi silencing.

PMID: 17553105 [PubMed - indexed for MEDLINE]

543: Planta. 2007 Sep;226(4):1007-16. Epub 2007 Jun 5.

Overexpression of OsCOIN, a putative cold inducible zinc finger protein,
increased tolerance to chilling, salt and drought, and enhanced proline level in
rice.

Liu K, Wang L, Xu Y, Chen N, Ma Q, Li F, Chong K.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, Chinese Academy of Science, Beijing 100093, China.

Rice (Oryza sativa L.) plant is sensitive to chilling, particularly at early
stages of seedling development. Here a novel cold-inducible gene, designated
OsCOIN (Oryza sativa cold-inducible), was isolated and characterized. Results
showed that OsCOIN protein, a RING finger protein, was localized in both nuclear
and cytoplasm membrane. OsCOIN is expressed in all rice organs and strongly
induced by low temperature, ABA, salt and drought. Over-expression of OsCOIN in
transgenic rice lines significantly enhanced their tolerance to cold, salt and
drought, accompanied by an up-regulation of OsP5CS expression and an increase of
cellular proline level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17549515 [PubMed - indexed for MEDLINE]

544: Plant Biotechnol J. 2007 Jul;5(4):537-53.

A potato carboxypeptidase inhibitor gene provides pathogen resistance in
transgenic rice.

Quilis J, Meynard D, Vila L, Avilö©s FX, Guiderdoni E, San Segundo B.

Consorcio CSIC-IRTA Laboratorio de Genö©tica Molecular Vegetal, Departamento de
Genö©tica Molecular, Instituto de Biologö­a Molecular de Barcelona, CSIC, Jordi
Girona 18, Barcelona, Spain.

A defensive role against insect attack has been traditionally attributed to plant
protease inhibitors. Here, evidence is described of the potential of a plant
protease inhibitor, the potato carboxypeptidase inhibitor (PCI), to provide
resistance to fungal pathogens when expressed in rice as a heterologous protein.
It is shown that rice plants constitutively expressing the pci gene exhibit
resistance against the economically important pathogens Magnaporthe oryzae and
Fusarium verticillioides. A M. oryzae carboxypeptidase was purified by affinity
chromatography and further characterized by mass spectrometry. This fungal
carboxypeptidase was found to be a novel carboxypeptidase B which was fully
inhibited by PCI. Overall, the results indicate that PCI exerts its antifungal
activity through the inhibition of this particular fungal carboxypeptidase B.
Although pci confers protection against fungal pathogens in transgenic rice, a
significant cost in insect resistance is observed. Thus, the weight gain of
larvae of the specialist insect Chilo suppressalis (striped stem borer) and the
polyphagous insect Spodoptera littoralis (Egyptian cotton worm) fed on pci rice
is significantly larger than that of insects fed on wild-type plants.
Homology-based modelling revealed structural similarities between the predicted
structure of the M. oryzae carboxypeptidase B and the crystal structure of insect
carboxypeptidases, indicating that PCI may function not only as an inhibitor of
fungal carboxypeptidases, but also as an inhibitor of insect carboxypeptidases.
The potential impact of the pci gene in terms of protection against fungal and
insect diseases is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17547659 [PubMed - indexed for MEDLINE]

545: Food Nutr Bull. 2006 Sep;27(3):265-6.

Quality protein maize.

Scrimshaw NS.

PMID: 17542118 [PubMed - indexed for MEDLINE]

546: Planta. 2007 Oct;226(5):1097-108. Epub 2007 Jun 1.

Overexpression of glycerol-3-phosphate acyltransferase gene improves chilling
tolerance in tomato.

Sui N, Li M, Zhao SJ, Li F, Liang H, Meng QW.

College of Life Sciences, Shandong Agricultural University, Key Lab of Crop
Biology of Shandong Province, Tai'an, 271018, People's Republic of China.

A tomato (Lycopersicon esculentum Mill.) glycerol-3-phosphate acyltransferase
gene (LeGPAT) was isolated. The deduced amino acid sequence revealed that LeGPAT
contained four acyltransferase domains, showing high identities with GPAT in
other plant species. A GFP fusion protein of LeGPAT was targeted to chloroplast
in cowpea mesophyll protoplast. RNA gel blot showed that the mRNA accumulation of
LeGPAT in the wild type (WT) was induced by chilling temperature. Higher
expression levels were observed when tomato leaves were exposed to 4 degrees C
for 4 h. RNA gel and western blot analysis confirmed that the sense gene LeGPAT
was transferred into the tomato genome and overexpressed under the control of
35S-CaMV. Although tomato is classified as a chilling-sensitive plant, LeGPAT
exhibited selectivity to 18:1 over 16:0. Overexpression of LeGPAT increased total
activity of LeGPAT and cis-unsaturated fatty acids in PG in thylakoid membrane.
Chilling treatment induced less ion leakage from the transgenic plants than from
the WT. The photosynthetic rate and the maximal photochemical efficiency of PS II
(Fv/Fm) in transgenic plants decreased more slowly during chilling stress and
recovered faster than in WT under optimal conditions. The oxidizable P700 in both
WT and transgenic plants decreased obviously at chilling temperature under low
irradiance, but the oxidizable P700 recovered faster in transgenic plants than in
the WT. These results indicate that overexpression of LeGPAT increased the levels
of PG cis-unsaturated fatty acids in thylakoid membrane, which was beneficial for
the recovery of chilling-induced PS I photoinhibition in tomato.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17541789 [PubMed - indexed for MEDLINE]

547: Transgenic Res. 2007 Oct;16(5):557-69. Epub 2007 May 31.

Pollen-mediated intraspecific gene flow from herbicide resistant oilseed rape
(Brassica napus L.).

Hö¼sken A, Dietz-Pfeilstetter A.

Institute for Plant Virology, Microbiology and Biosafety, Federal Biological
Research Centre for Agriculture and Forestry, Messeweg 11-12, 38104 Braunschweig,
Germany.

The cultivation of genetically modified (GM) herbicide resistant oilseed rape
(Brassica napus) has increased over the past few years. The transfer of herbicide
resistance genes via pollen (gene flow) from GM crops to non-GM crops is of
relevance for the realisation of co-existence of different agricultural
cultivation forms as well as for weed management. Therefore the likelihood of
pollen-mediated gene flow has been investigated in numerous studies. Despite the
difficulty to compare different experiments with varying levels of outcrossing,
we performed a literature search for world-wide studies on cross-fertilisation in
fully fertile oilseed rape. The occurrence and frequency of pollen-mediated
intraspecific gene flow (outcrossing rate) can vary according to cultivar,
experimental design, local topography and environmental conditions. The
outcrossing rate from one field to another depends also on the size and
arrangement of donor and recipient populations and on the ratio between donor and
recipient plot size. The outcrossing levels specified in the presented studies
are derived mostly from experiments where the recipient field is either
surrounding the donor field (continuous design) or is located as a patch at
different distances from the donor field (discontinuous design). Reports of gene
flow in Brassica napus generally show that the amount of cross-fertilisation
decreases as the distance from the pollen source increases. The evidence given in
various studies reveals that the bulk of GM cross-fertilisation occurs within the
first 10 m of the recipient field. The removal of the first 10 m of a
non-transgenic field facing a GM crop might therefore be more efficient for
reducing the total level of cross-fertilisation in a recipient sink population
than to recommend separation distances. Future experiments should investigate
cross-fertilisation with multiple adjacent donor fields at the landscape level
under different spatial distributions of rapeseed cultivars and different
cropping systems. The level of cross-fertilisation occurring over the whole field
is mainly important for co-existence and has not been investigated in
agricultural scale experiments until now. Potential problems with herbicide
resistant oilseed rape volunteers arising from intraspecific gene flow can be
largely solved by the choice of suitable cultivars and herbicides as well as by
soil management.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17541721 [PubMed - indexed for MEDLINE]

548: Plant Cell. 2007 May;19(5):1433-4. Epub 2007 May 31.

The freedom to innovate: a privilege or a right?

Jorgensen R.

Publication Types:
Editorial

PMID: 17540714 [PubMed - indexed for MEDLINE]

549: Environ Entomol. 2007 Jun;36(3):646-54.

Arthropod abundance and diversity in Bt and non-Bt rice fields.

Li FF, Ye GY, Wu Q, Peng YF, Chen XX.

State Key Laboratory of Rice Biology, Institute of Insect Sciences, Zhejiang
University, Hangzhou 310029, China.

In a field experiment, possible effects of transgenic Bt rice on arthropod
communities under paddy field conditions were assessed for 3 yr in terms of
arthropod guild dominance, family composition, dominance distribution of each
guild, individuals of each guild, and community indices (including Shannon-Weaver
diversity index and dominant concentration index). Our results overall suggested
no significant differences between the Bt and control rice plots in these
arthropod community-specific parameters. The similarity of arthropod communities
in the Bt and control rice plots was apparently high. Based on our findings, we
conclude that Bt rice generally exerts no marked negative effects on the
arthropod community in paddy fields.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17540077 [PubMed - indexed for MEDLINE]

550: J Biosci. 2007 Apr;32(3):621-8.

Functional validation of a novel isoform of Na+/H+ antiporter from Pennisetum
glaucum for enhancing salinity tolerance in rice.

Verma D, Singla-Pareek SL, Rajagopal D, Reddy MK, Sopory SK.

Plant Molecular Biology, International Center for Genetic Engineering and
Biotechnology, Aruna Asaf Ali Marg, New Delhi 10067, India.

Salt stress is an environmental factor that severely impairs plant growth and
productivity. We have cloned a novel isoform of a vacuolar Na+/H+ antiporter from
Pennisetum glaucum (PgNHX1) that contains 5 transmembrane domains in contrast to
AtNHX1 and OsNHX1 which have 9 transmembrane domains. Recently we have shown that
PgNHX1 could confer high level of salinity tolerance when overexpressed in
Brassica juncea. Here,we report the functional validation of this antiporter in
crop plant rice.Overexpression of PgNHX1 conferred high level of salinity
tolerance in rice. Transgenic rice plants overexpressing PgNHX1 developed more
extensive root system and completed their life cycle by setting flowers and seeds
in the presence of 150 mM NaCl. Our data demonstrate the potential of PgNHX1 for
imparting enhanced salt tolerance capabilities to salt-sensitive crop plants for
growing in high saline areas.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17536181 [PubMed - indexed for MEDLINE]

551: Plant Cell Rep. 2007 Sep;26(9):1575-84. Epub 2007 May 30.

Analysis of the limitations of hepatitis B surface antigen expression in soybean
cell suspension cultures.

Ganapathi TR, Sunil Kumar GB, Srinivas L, Revathi CJ, Bapat VA.

Plant Cell culture Technology Section, Nuclear Agriculture and Biotechnology
Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.

Soybean cell suspension cultures were transformed using Agrobacterium tumefaciens
harboring pHBS/pHER constructs to express hepatitis B surface antigen (HBsAg).
The transformed colonies were selected and analyzed for the expression of HBsAg
by PCR, reverse transcription (RT) PCR, Western blot and ELISA analysis. The
maximum expression of 700 ng/g F.W. was noted in pHER transformed cells. The
highest expressing colonies were used to initiate the cell suspension cultures
and the expression of HBsAg was estimated periodically. The expression levels
were reduced drastically in cell suspension cultures compared to the colonies
maintained on semi-solid medium. Various parameters were studied to maximize the
cell growth and to retain the expression levels. The supplementation of culture
medium with a protease inhibitor, leupeptin hemisulfate could restore up to 50%
of HBsAg expression in cell suspension cultures. This is the first report to
investigate the possible cause and solution to the loss of recombinant protein
expression levels in plant cell suspension cultures.

PMID: 17534624 [PubMed - indexed for MEDLINE]

552: Theor Appl Genet. 2007 Aug;115(4):571-90. Epub 2007 May 30.

Erratum in:
Theor Appl Genet. 2007 Aug;115(4):591.

Complete chloroplast genome sequences of Hordeum vulgare, Sorghum bicolor and
Agrostis stolonifera, and comparative analyses with other grass genomes.

Saski C, Lee SB, Fjellheim S, Guda C, Jansen RK, Luo H, Tomkins J, Rognli OA,
Daniell H, Clarke JL.

Clemson University Genomics Institute, Clemson University, Biosystems Research
Complex, 51 New Cherry Street, Clemson, SC 29634, USA.

Comparisons of complete chloroplast genome sequences of Hordeum vulgare, Sorghum
bicolor and Agrostis stolonifera to six published grass chloroplast genomes
reveal that gene content and order are similar but two microstructural changes
have occurred. First, the expansion of the IR at the SSC/IRa boundary that
duplicates a portion of the 5' end of ndhH is restricted to the three genera of
the subfamily Pooideae (Agrostis, Hordeum and Triticum). Second, a 6 bp deletion
in ndhK is shared by Agrostis, Hordeum, Oryza and Triticum, and this event
supports the sister relationship between the subfamilies Erhartoideae and
Pooideae. Repeat analysis identified 19-37 direct and inverted repeats 30 bp or
longer with a sequence identity of at least 90%. Seventeen of the 26 shared
repeats are found in all the grass chloroplast genomes examined and are located
in the same genes or intergenic spacer (IGS) regions. Examination of simple
sequence repeats (SSRs) identified 16-21 potential polymorphic SSRs. Five IGS
regions have 100% sequence identity among Zea mays, Saccharum officinarum and
Sorghum bicolor, whereas no spacer regions were identical among Oryza sativa,
Triticum aestivum, H. vulgare and A. stolonifera despite their close phylogenetic
relationship. Alignment of EST sequences and DNA coding sequences identified six
C-U conversions in both Sorghum bicolor and H. vulgare but only one in A.
stolonifera. Phylogenetic trees based on DNA sequences of 61 protein-coding genes
of 38 taxa using both maximum parsimony and likelihood methods provide moderate
support for a sister relationship between the subfamilies Erhartoideae and
Pooideae.

Publication Types:
Comparative Study
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17534593 [PubMed - indexed for MEDLINE]

553: Pest Manag Sci. 2007 Jul;63(7):658-76.

Mites for the control of pests in protected cultivation.

Gerson U, Weintraub PG.

Department of Entomology, Faculty of Agricultural, Food and Environmental Quality
Sciences, Rehovot 76100, Israel.

The production of crops under protected conditions is increasing worldwide. Owing
to growing consumer demands for healthy and green produce, and intensifying
pesticide resistance, non-chemical solutions--foremost among which is biological
control--are being sought. The authors review recent advances related to the
application of predatory mites for the control of greenhouse pests, and discuss
interactions among acarine biocontrol agents (ABAs) and the effects of crop
plants and new technologies on ABAs, such as artificial lighting, elevated carbon
dioxide levels and genetically modified organisms. This is followed by a
discussion of the problems associated with the search for and use of new ABAs,
including management, the benefits of modelling and avenues of future research.
Copyright (c) 2007 Society of Chemical Industry.

Publication Types:
Review

PMID: 17533640 [PubMed - indexed for MEDLINE]

554: Plant Cell Physiol. 2007 Jul;48(7):958-70. Epub 2007 May 26.

Isolation and functional analysis of a MYB transcription factor gene that is a
key regulator for the development of red coloration in apple skin.

Ban Y, Honda C, Hatsuyama Y, Igarashi M, Bessho H, Moriguchi T.

Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1
Tennoudai, Tsukuba, Ibaraki, 305-8572 Japan.

Red coloration of apple (Malus x domestica) skin is an important determinant of
consumer preference and marketability. Anthocyanins are responsible for this
coloration, and their accumulation is positively correlated with the expression
level of anthocyanin biosynthetic genes. Regulation of expression of these genes
is believed to be controlled by MYB transcription factors, and the MYB
transcription factors involved in the activation of anthocyanin biosynthetic
genes have been isolated in various plants. In the present study, we isolated and
characterized a MYB transcription factor gene (MdMYBA) from apple skin.
Characterization of MdMYBA demonstrated that (i) MdMYBA expression was
specifically regulated depending on the tissue and cultivar/species; (ii) its
expression level was much higher in a deep-red cultivar ('Jonathan') than in a
pale-red cultivar ('Tsugaru'); (iii) when cauliflower mosaic virus 35S::MdMYBA
was introduced into the cotyledons of apple seedlings by means of a transient
assay, reddish-purple spots were induced, and MdMYBA also induced anthocyanin
accumulation in reproductive tissues of transgenic tobacco; (iv) the expression
of MdMYBA was induced by UV-B irradiation and low-temperature treatment, both of
which are known to be important in the promotion of anthocyanin accumulation in
apple skin; (v) MdMYBA bound specifically to an anthocyanidin synthase (MdANS)
promoter region in a gel-shift assay; and (vi) MdMYBA was mapped to the near
region of the BC226-STS (a1) marker for the red skin color locus (R(f)). These
results suggest that MdMYBA is a key regulatory gene in anthocyanin biosynthesis
in apple skin.

PMID: 17526919 [PubMed - indexed for MEDLINE]

555: Plant Cell. 2007 May;19(5):1580-9. Epub 2007 May 25.

Structure and function of a mitochondrial late embryogenesis abundant protein are
revealed by desiccation.

Tolleter D, Jaquinod M, Mangavel C, Passirani C, Saulnier P, Manon S, Teyssier E,
Payet N, Avelange-Macherel MH, Macherel D.

Unitö© Mixte de Recherche 1191, Physiologie Molö©culaire des Semences, Universitö©
d'Angers/Institut National d'Horticulture/Institut National de la Recherche
Agronomique, Angers F-49045, France.

Few organisms are able to withstand desiccation stress; however, desiccation
tolerance is widespread among plant seeds. Survival without water relies on an
array of mechanisms, including the accumulation of stress proteins such as the
late embryogenesis abundant (LEA) proteins. These hydrophilic proteins are
prominent in plant seeds but also found in desiccation-tolerant organisms. In
spite of many theories and observations, LEA protein function remains unclear.
Here, we show that LEAM, a mitochondrial LEA protein expressed in seeds, is a
natively unfolded protein, which reversibly folds into alpha-helices upon
desiccation. Structural modeling revealed an analogy with class A amphipathic
helices of apolipoproteins that coat low-density lipoprotein particles in
mammals. LEAM appears spontaneously modified by deamidation and oxidation of
several residues that contribute to its structural features. LEAM interacts with
membranes in the dry state and protects liposomes subjected to drying. The
overall results provide strong evidence that LEAM protects the inner
mitochondrial membrane during desiccation. According to sequence analyses of
several homologous proteins from various desiccation-tolerant organisms, a
similar protection mechanism likely acts with other types of cellular membranes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17526751 [PubMed - indexed for MEDLINE]

556: Biotechnol J. 2007 Jul;2(7):826-32.

Safety assessment of genetically modified organisms of plant origin in the
Russian Federation.

Tyshko NV, Aksyuk IN, Tutelyan VA.

Institute of Nutrition, Russian Academy of Medical Sciences, Moscow, Russian
Federation.

The beginning of the 21st century is characterized by growing interest in the
problems of biosafety, which are determined, on the one hand, by the wide use of
novel biotechnologies and the necessity to develop the adequate precautionary
measures, and, on the other hand, by the objective threat of bioterrorism.
Therefore, improvement of the estimation system for genetically modified (GM)
sources of food and strengthening the control of their circulation are the urgent
problems of modern biology and medicine. Russia is one of the countries where the
estimation system of food products obtained from the GM sources is rather
efficient. The key features of this system are the complex toxicological and
epidemiological examinations. One of the main parts of GM food safety assessment
is based upon detection of their potentially toxic properties, which could
provoke unintended effects of the genetic modification.

Publication Types:
Review

PMID: 17526054 [PubMed - indexed for MEDLINE]

557: Science. 2007 May 25;316(5828):1185-8.

Comment in:
Science. 2007 May 25;316(5828):1114-7.

Dicamba resistance: enlarging and preserving biotechnology-based weed management
strategies.

Behrens MR, Mutlu N, Chakraborty S, Dumitru R, Jiang WZ, Lavallee BJ, Herman PL,
Clemente TE, Weeks DP.

Department of Biochemistry, University of Nebraska, Lincoln, NE 68588, USA.

The advent of biotechnology-derived, herbicide-resistant crops has revolutionized
farming practices in many countries. Facile, highly effective, environmentally
sound, and profitable weed control methods have been rapidly adopted by crop
producers who value the benefits associated with biotechnology-derived weed
management traits. But a rapid rise in the populations of several troublesome
weeds that are tolerant or resistant to herbicides currently used in conjunction
with herbicide-resistant crops may signify that the useful lifetime of these
economically important weed management traits will be cut short. We describe the
development of soybean and other broadleaf plant species resistant to dicamba, a
widely used, inexpensive, and environmentally safe herbicide. The dicamba
resistance technology will augment current herbicide resistance technologies and
extend their effective lifetime. Attributes of both nuclear- and
chloroplast-encoded dicamba resistance genes that affect the potency and expected
durability of the herbicide resistance trait are examined.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17525337 [PubMed - indexed for MEDLINE]

558: Science. 2007 May 25;316(5828):1116-7.

Agbiotech. Glyphosate--the conservationist's friend?

Service RF.

Publication Types:
News

PMID: 17525313 [PubMed - indexed for MEDLINE]

559: Science. 2007 May 25;316(5828):1114-7.

Comment on:
Science. 2007 May 25;316(5828):1185-8.

Agbiotech. A growing threat down on the farm.

Service RF.

Publication Types:
Comment
News

PMID: 17525312 [PubMed - indexed for MEDLINE]

560: Plant Physiol Biochem. 2007 May;45(5):270-6. Epub 2007 Apr 6.

Interspecies compatibility of NAS1 gene promoters.

Ito S, Inoue H, Kobayashi T, Yoshiba M, Mori S, Nishizawa N, Higuchi K.

Laboratory of Plant Production Chemistry, Department of Applied Biological
Chemistry, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo
156-8502, Japan.

Nicotianamine and nicotianamine synthase (NAS) play key roles in iron nutrition
in all higher plants. However, the mechanism underlying the regulation of NAS
expression differs among plant species. Sequences homologous to iron
deficiency-responsive elements (IDEs), i.e., cis-acting elements, are found on
the promoters of these genes. We aimed to verify the interspecies compatibility
of the Fe-deficiency response of NAS1 genes and understand the universal
mechanisms that regulate their expression patterns in higher plants. Therefore,
we introduced the graminaceous (Hordeum vulgare L. and Oryza sativa L.) NAS1
promoter::GUS into dicots (Nicotiana tabacum L. and Arabidopsis thaliana L.). Fe
deficiency induced HvNAS1 expression in the shoots and roots when introduced into
rice. HvNAS1 promoter::GUS and OsNAS1 promoter::GUS induced strong expression of
GUS under Fe-deficient conditions in transformed tobacco. In contrast, these
promoters only definitely functioned in Arabidopsis transformants. These results
suggest that some Fe nutrition-related trans-factors are not compatible between
graminaceous plants and Arabidopsis. HvNAS1 promoter::GUS induced GUS activity
only in the roots of transformed tobacco under Fe-deficient conditions. On the
other hand, OsNAS1 promoter::GUS induced GUS activity in both the roots and
shoots of transformed tobacco under conditions of Fe deficiency. In tobacco
transformants, the induction of GUS activity was induced earlier in the shoots
than roots. These results suggest that the HvNAS1 and OsNAS1 promoters are
compatible with Fe-acquisition-related trans-factors in the roots of tobacco and
that the OsNAS1 promoter is also compatible with some shoot-specific Fe
deficiency-related trans-factors in tobacco.

PMID: 17524656 [PubMed - indexed for MEDLINE]

561: Phytochemistry. 2007 Jun;68(12):1632-41. Epub 2007 May 23.

The rice (E)-beta-caryophyllene synthase (OsTPS3) accounts for the major
inducible volatile sesquiterpenes.

Cheng AX, Xiang CY, Li JX, Yang CQ, Hu WL, Wang LJ, Lou YG, Chen XY.

National Key Laboratory of Plant Molecular Genetics, Institute of Plant
Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences, Shanghai 200032, China.

Terpenoids serve as both constitutive and inducible defense chemicals in many
plant species, and volatile terpenes participate in plant a indirect defense by
attracting natural enemies of the herbivores. The rice (Oryza sativa L.) genome
contains about 50 genes encoding putative terpene synthases (TPSs). Here we
report that two of the rice sesquiterpene synthase genes, OsTPS3 and OsTPS13,
encode (E)-beta-caryophyllene synthase and (E,E)-farnesol synthase, respectively.
In vitro, the recombinant protein of OsTPS3 catalyzed formation of
(E)-beta-caryophyllene and several other sesquiterpenes, including beta-elemene
and alpha-humulene, all being components of inducible volatiles of rice plants.
The transcript levels of OsTPS3 exhibit a circadian rhythm of fluctuation, and
its expression was also greatly induced by methyl jasmonate (MeJA). In addition,
expression of OsTPS3 in transgenic plants of Arabidopsis thaliana resulted in
emitting high quantities of OsTPS3 products. We also overexpressed OsTPS3 in rice
plants which then produced more (E)-beta-caryophyllene after MeJA treatment.
Finally, we found that the MeJA-treated transgenic rice plants attracted more
parasitoid wasps of Anagrus nilaparvatae than the wild-type. These results
demonstrate that OsTPS3, an enzyme catalyzing the formation of volatile
sesquiterpenes, plays a role in indirect defense of rice plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17524436 [PubMed - indexed for MEDLINE]

562: Bull Entomol Res. 2007 Jun;97(3):265-80.

Statistical models to evaluate invertebrate-plant trophic interactions in arable
systems.

Bohan DA, Hawes C, Haughton AJ, Denholm I, Champion GT, Perry JN, Clark SJ.

Rothamsted Research, Harpenden, Herts, AL5 2JQ, UK. David.Bohan@bbrsc.ac.uk

Over the past 40 years there have been marked shifts in arable farmland
management that are widely believed to have had a considerable impact on
flowering plants and invertebrates and the small mammals and birds that rely upon
them. It is not yet possible to predict the dynamics of plants and invertebrates
either with past or future changes in farmland management. This study
investigates whether a basic invertebrate classification, formed of broad trophic
groups, can be used to describe interactions between invertebrates and their
resource plants and evaluate management impacts for genetically modified,
herbicide-tolerant (GMHT) and conventional herbicide management in both spring-
and winter-sown oilseed rape. It is argued that the analyses validate
trophic-based approaches for describing the dynamics of invertebrates in farmland
and that linear models might be used to describe the changes in invertebrate
trophic group abundance in farmland when driven by primary producer abundance or
biomass and interactions between invertebrates themselves. The analyses indicate
that invertebrate dynamics under GMHT management are not unique, but similar to
conventional management occurring over different resource ranges, and that
dynamics differed considerably between spring- and winter-sown oilseed rape.
Thus, herbicide management was of much lower impact on trophic relationships than
sowing date. Results indicate that invertebrate dynamics in oilseed rape are
regulated by a combination of top-down and bottom-up trophic processes.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 17524158 [PubMed - indexed for MEDLINE]

563: Anal Chem. 2007 Jul 1;79(13):5071-7. Epub 2007 May 25.

Analysis of chiral amino acids in conventional and transgenic maize.

Herrero M, Ibö¡ö±ez E, Martö­n-Alvarez PJ, Cifuentes A.

Institute of Industrial Fermentations (CSIC), Juan de la Cierva 3, 28006 Madrid,
Spain.

In this work, a new chiral micellar electrokinetic chromatography with
laser-induced fluorescence detection (chiral-MEKC-LIF) method is proposed to
identify and quantify D- and L-amino acids in three lines of transgenic maize and
their corresponding nontransgenic parental lines grown under identical
conditions. The optimized procedure includes amino acids extraction,
derivatization with FITC and chiral-MEKC-LIF separation in a background
electrolyte composed of 100 mM sodium tetraborate, 80 mM SDS, and 20 mM beta-CD
at pH 10.0. The D- and L-forms of Arg, Ser, Ala, Glu, and Asp, corresponding to
the majority amino acids usually found in maize, are separated in less than 25
min with efficiencies up to 890,000 plates/m and high sensitivity (i.e., LODs as
low as 160 nM were obtained for D-Arg for a signal-to-noise ratio of three),
allowing the detection of 1% D-Arg in the presence of 99% of its opposite
enantiomer. Using this method, different D-amino acids are detected in all
investigated maize samples providing the reproducible quantification of the
D-enantiomeric excess (% d-aa) for each amino acid calculated as % D-aa =
100D-aa/(D-aa + L-aa). Thus, significant differences were observed among the %
d-aa values for the different conventional varieties (Aristis, Tietar, and
PR33P66 maize) as could be expected from their natural variability. More
interestingly, comparing each conventional maize with its corresponding
transgenic line, very similar % D-aa values were obtained for one of the studied
maize couples (Tietar vs Tietar-Bt) what could be presented as a new proof of
their substantial equivalence. However, significant differences in the % d-aa
values were observed for the other lines of maize studied. It is concluded that
enantioselective procedures can open new perspectives in the study of transgenic
organisms in order to corroborate (or not) the equivalence with their
conventional counterparts.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17523597 [PubMed - indexed for MEDLINE]

564: Planta. 2007 Aug;226(3):773-83. Epub 2007 May 24.

Developmental control of Arabidopsis seed oil biosynthesis.

Wang H, Guo J, Lambert KN, Lin Y.

Department of Crop Sciences, University of Illinois, Urbana, IL 61801, USA.

Arabidopsis transcriptional factors LEAFY COTYLEDON1 (LEC1), LEAFY COTYLEDON2
(LEC2), FUSCA3 (FUS3), ABSCISIC ACID3 (ABI3), and ABSCISIC ACID5 (ABI5) are known
to regulate multiple aspects of seed development. In an attempt to understand the
developmental control of storage product accumulation, we observed the expression
time course of the five transcripts. The sequential expression of these factors
during seed fill suggests differentiation of their normal responsibilities. By
extending the expression periods of the two early genes LEC1 and LEC2 in
transgenic seeds, we demonstrated that the subsequent timing of FUS3, ABI3, and
ABI5 transcripts depends on LEC1 and LEC2. Because a delayed onset or reduced
level of FUS3 mRNA coincided with reduction of seed oil content in the transgenic
seeds, the role of FUS3 in oil deposition was further examined. Analysis of
published seed transcriptome data indicated that FUS3 transcript increased
together with nearly all the plastidial fatty acid biosynthetic transcripts
during development. The ability of FUS3 to rapidly induce fatty acid biosynthetic
gene expression was confirmed using transgenic Arabidopsis seedlings expressing a
dexamethasone (DEX)-inducible FUS3 and Arabidopsis mesophyll protoplasts
transiently expressing the FUS3 gene. By accommodating the current evidence, we
propose a hierarchical architecture of the transcriptional network in Arabidopsis
seeds in which the oil biosynthetic pathway is integrated through the master
transcriptional factor FUS3.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17522888 [PubMed - indexed for MEDLINE]

565: Adv Biochem Eng Biotechnol. 2007;107:235-78.

Ecological impacts of genetically modified crops: ten years of field research and
commercial cultivation.

Sanvido O, Romeis J, Bigler F.

Agroscope Reckenholz-Tö¤nikon Research Station ART, Reckenholzstr. 191, 8046,
Zurich, Switzerland. olivier.sanvido@art.admin.ch

The worldwide commercial cultivation of genetically modified (GM) crops has
raised concerns about potential adverse effects on the environment resulting from
the use of these crops. Consequently, the risks of GM crops for the environment,
and especially for biodiversity, have been extensively assessed before and during
their commercial cultivation. Substantial scientific data on the environmental
effects of the currently commercialized GM crops are available today. We have
reviewed this scientific knowledge derived from the past 10 years of worldwide
experimental field research and commercial cultivation. The review focuses on the
currently commercially available GM crops that could be relevant for agriculture
in Western and Central Europe (i.e., maize, oilseed rape, and soybean), and on
the two main GM traits that are currently commercialized, herbicide tolerance
(HT) and insect resistance (IR). The sources of information included
peer-reviewed scientific journals, scientific books, reports from regions with
extensive GM crop cultivation, as well as reports from international governmental
organizations. The data available so far provide no scientific evidence that the
cultivation of the presently commercialized GM crops has caused environmental
harm. Nevertheless, a number of issues related to the interpretation of
scientific data on effects of GM crops on the environment are debated
controversially. The present review highlights these scientific debates and
discusses the effects of GM crop cultivation on the environment considering the
impacts caused by cultivation practices of modern agricultural systems.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17522828 [PubMed - indexed for MEDLINE]

566: Adv Biochem Eng Biotechnol. 2007;107:207-34.

Assessing effects of transgenic crops on soil microbial communities.

Widmer F.

Molecular Ecology, Agroscope Reckenholz-Tö¤nikon Research Station ART,
Reckenholzstrasse 191, 8046, Zö¼rich, Switzerland. franco.widmer@art.admin.ch

Deleterious effects of transgenic plants on soils represent an often expressed
concern, which has catalyzed numerous studies in the recent past. In this
literature review, studies addressing this question have been compiled. A total
of 60 studies has been found, and their findings as well as their analytical
approaches are summarized. These studies analyzed the effects of seven different
types of genetically engineered traits, i.e., herbicide tolerance, insect
resistance, virus resistance, proteinase inhibitors, antimicrobial activity,
environmental application, and biomolecule production. Sixteen genetically
engineered plant species were investigated in these studies including corn,
canola, soybean, cotton, potato, tobacco, alfalfa, wheat, rice, tomato, papaya,
aubergine, and silver birch. Many of these plants and traits have not been
commercialized and represent experimental model systems. Effects on soil
microbial characteristics have been described in various studies, indicating the
sensitivity and feasibility of the analytical approaches applied. However,
classification of the observed effects into acceptable and unacceptable ones has
not been possible so far. Establishment of validated indicators for adverse
effects represents a scientific challenge for the near future, and will assist
risk assessment and regulation of transgenic plants commercially released to the
field.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17522827 [PubMed - indexed for MEDLINE]

567: Adv Biochem Eng Biotechnol. 2007;107:173-205.

Genetic and ecological consequences of transgene flow to the wild flora.

Felber F, Kozlowski G, Arrigo N, Guadagnuolo R.

Laboratoire de Botanique ö©volutive, Institut de Biologie, Universitö© de
Neuchö¢tel, rue Emile-Argand 11, 2009, Neuchö¢tel, Switzerland.
Francois.Felber@unine.ch

Gene flow from crops to wild relatives by sexual reproduction is one of the major
issues in risk assessment for the cultivation of genetically engineered (GE)
plants. The main factors which influence hybridization and introgression, the two
processes of gene flow, as well as the accompanying containment measures of the
transgene, are reviewed. The comparison of risks between Switzerland and Europe
highlights the importance of regional studies. Differences were assessed for
barley, beet and wheat. Moreover, transgene flow through several wild species
acting as bridge (bridge species) has been up to now poorly investigated. Indeed,
transgene flow may go beyond the closest wild relative, as in nature several wild
species complexes hybridize. Its importance is assessed by several examples in
Poaceae. Finally, the transgene itself has genetic and ecological consequences
that are reviewed. Transgenic hybrids between crops and wild relatives may have
lower fitness than the wild relatives, but in several cases, no cost was
detected. On the other hand, the transgene provides advantages to the hybrids, in
the case of selective value as a Bt transgene in the presence of herbivores.
Genetic and ecological consequences of a transgene in a wild species are complex
and depend on the type of transgene, its insertion site, the density of plants
and ecological factors. More studies are needed for understanding the short and
long term consequences of escape of a transgene in the wild.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17522826 [PubMed - indexed for MEDLINE]

568: Adv Biochem Eng Biotechnol. 2007;107:133-51.

Prospects for biopolymer production in plants.

van Beilen JB, Poirier Y.

Dö©partement de Biologie Molö©culaire Vö©gö©tale, Universitö© de Lausanne, Bö¢timent
Biophore, 1015, Lausanne, Switzerland.

It is likely that during this century polymers based on renewable materials will
gradually replace industrial polymers based on petrochemicals. This chapter gives
an overview of the current status of research on plant biopolymers that are used
as a material in non-food applications. We cover technical and scientific
bottlenecks in the production of novel or improved materials, and the potential
of using transgenic or alternative crops in overcoming these bottlenecks. Four
classes of biopolymers will be discussed: starch, proteins, natural rubber, and
poly-beta-hydroxyalkanoates. Renewable polymers produced by chemical
polymerization of monomers derived from sugars, vegetable oil, or proteins, are
not considered here.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17522824 [PubMed - indexed for MEDLINE]

569: Adv Biochem Eng Biotechnol. 2007;107:97-112.

Exploration and Swiss field-testing of a viral gene for specific quantitative
resistance against smuts and bunts in wheat.

Schlaich T, Urbaniak B, Plissonnier ML, Malgras N, Sautter C.

Institute of Plant Science, Swiss Federal Institute of Technology Zurich,
Universitö¤tsstr. 2, 8092, Zurich, Switzerland. christof.sautter@ipw.biol.ethz.ch

The viral gene for the killer protein 4 (KP4) has been explored for its
antifungal effect in genetically modified wheat to defeat specifically the
seed-transmitted smut and bunt diseases. In vitro both important seed-transmitted
diseases of wheat, loose smut (Ustilago tritici) and stinking smut (Tilletia
caries), are susceptible to KP4, whereas all other organisms tested so far proved
to be not susceptible to KP4. For studies in planta we used stinking smut as a
model fungus. In greenhouse experiments, two KP4-transgenic wheat lines showed up
to 30% lower symptom development as compared to the nontransgenic control. As the
last step in the proof of concept, field-testing has shown for the first time
increased fungal resistance of a transgene in wheat. Due to its specificity
against smuts and bunts, KP4 presents a very low risk to humans and the
environment. Field-testing in Switzerland is regulated by a strong law, which for
research is acceptable if legally and scientifically correctly applied.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17522822 [PubMed - indexed for MEDLINE]

570: Adv Biochem Eng Biotechnol. 2007;107:57-68.

Genetically modified organisms in the United States: implementation, concerns,
and public perception.

Oeschger MP, Silva CE.

Department of Microbiology, Immunology and Parasitology, Louisiana State
University Health Sciences Center, New Orleans, LA 70112, USA. moesch@lsuhsc.edu

We examine the state of biotechnology with respect to genetically modified (GM)
organisms in agriculture. Our focus is on the USA, where there has been
significant progress and implementation but where, to date, the matter has drawn
little attention. GM organisms are the result of lateral gene transfers, the
transfer of genes from one species to another, or sometimes, from one kingdom to
another. The introduction of foreign genes makes some people very uncomfortable,
and a small group of activists have grave concerns about the technology. Attempts
by activists to build concern in the general public have garnered little
attention; however, the producers of GM organisms have responded to their
concerns and established extensive testing programs to be applied to each
candidate organism that is produced. In the meantime, GM varieties of corn,
cotton, soybean and rapeseed have been put into agricultural production and are
now extensively planted. These crops, and the other, newer GM crops, have
produced no problems and have pioneered a silent agricultural revolution in the
USA.

Publication Types:
Review

PMID: 17522820 [PubMed - indexed for MEDLINE]

571: Adv Biochem Eng Biotechnol. 2007;107:1-11.

The gap between science and perception: the case of plant biotechnology in
Europe.

Einsele A.

Internutrition, Postfach, 8035, Zurich, Switzerland.
arthur.einsele@internutrition.ch

Although the global area of biotech crops continues to climb for the tenth
consecutive year at a sustainable double-digit growth rate, the acceptance of
biotech products from agriculture in Europe is still low. There is a gap between
science and perception. It is a strong belief that the public turning against
science and against GM food has been encouraged by the negative activities of NGO
groups. Scientists have to overcome the purely risk-based discussion, and the
benefits of plant biotechnology have to be made literally visible. GM food should
be available, the benefits should be tangible and the consumer should have fun
with such novel food. The gap could be reduced if genetically modified plants and
the products thereof were regulated in the same way as classical products.

Publication Types:
Review

PMID: 17522817 [PubMed - indexed for MEDLINE]

572: Biochem Biophys Res Commun. 2007 Jul 13;358(4):983-9. Epub 2007 May 11.

Overproduction of OsSLRL2 alters the development of transgenic Arabidopsis
plants.

Liu T, Gu JY, Xu CJ, Gao Y, An CC.

The National Laboratory of Protein Engineering and Plant Genetic Engineering,
College of Life Sciences, Peking University, Beijing 100871, PR China.

SLR1 (SLENDER RICE 1) was thought to be the sole DELLA protein in rice
considering the constitutive GA response phenotype of slr1 mutants. There were
two other SLR1 homologous SLRL1 and SLRL2 (SLR1 like 1 and 2) which did not have
DELLA domain but still shared high level similarity to the C-terminal region of
SLR1 found after searching the whole rice genome. SLRL2 specially expressed in
the embryo of immature rice seeds and the expression of SLRL2 was increased when
treated with GA(3). The SLRL2 over-expressed transgenic Arabidopsis plants were
semi-dwarfed, late flowering, and insensitive to GA. Moreover, the expression of
AtGA20ox1 and AtGA3ox1 was increased and the expression of AtGA2ox1 decreased,
indicating SLRL2 was a repressor of GA signaling. We suggested SLRL2 might
function to overcome too strong GA responses and maintained a basic repression.
Furthermore, a different form of DELLA family in monocots against dicots was
discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17521606 [PubMed - indexed for MEDLINE]

573: Clin Exp Allergy. 2007 Jun;37(6):918-28.

Genetically glycosylated ovomucoid third domain can modulate Immunoglobulin E
antibody production and cytokine response in BALB/c mice.

Rupa P, Nakamura S, Mine Y.

Department of Food Science, University of Guelph, Guelph, ON, Canada N1G 2W1.

BACKGROUND: Food allergies are on the rise and it is estimated that in North
America, 8% of the children and 4% of the adults have food allergies. Food
allergies tend to occur more often in children than in adults due to their
immature digestive and immune systems. Hen's egg is among the most common cause
of food-induced allergic reactions in North America. OBJECTIVE: The present study
was undertaken to investigate the role of N-glycans of the third domain of
ovomucoid in IgE binding and modulation of allergen-specific immune response in
BALB/c mice. METHODS: The cDNA encoding the third domain of ovomucoid was
inserted into the yeast genome and expressed in Pichia pastoris X-33 cells, under
the control of the glyceraldehyde-3-phosphate (GAP) dehydrogenase promoter for
constitutive expression to obtain a post-translationally modified and
functionally active ovomucoid third domain. Upon expression, the protein was
secreted into the extracellular medium and was purified by size exclusion
chromatography. The recombinant protein was produced at 10 mg/L of the culture
supernatant. BALB/c mice were sensitized with the recombinant and native forms of
glycosylated ovomucoid third domain antigen. The allergic response of the native
and the recombinant glycosylated forms of ovomucoid third domain antigens were
compared using antibody and cytokine measurements. RESULTS: ELISA tests indicated
a significant decrease in specific IgE antibodies to the recombinant N-linked
glycosylated form (P-Gly), when compared with the native glycosylated form
(DIII+) using mice sera. Immunization with P-Gly induced the production of
IFN-gamma [T-helper type 1 (Th1) response] and lowered the production of IL-4
(Th2 response), and a skewed balance towards the Th1 cytokine demonstrated that
P-Gly has a modulating ability on Th1/Th2 balance to down-regulate Th2 response.
Furthermore, N-linked glycan (N28) in the third domain of ovomucoid was shown to
be associated with suppression of the allergic response. CONCLUSION: Therefore,
we can conclude that P-Gly facilitates and contributes to the discovery of new
molecular target for the development of a safe and specific therapeutic vaccine
for the treatment of egg allergy, and oligosaccharides do seem to play a major
role in the suppression of IgE-binding activity.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17517106 [PubMed - indexed for MEDLINE]

574: Prep Biochem Biotechnol. 2007;37(3):239-46.

Growth and rutin production in hairy root cultures of buckwheat (Fagopyrum
esculentum M.).

Lee SY, Cho SI, Park MH, Kim YK, Choi JE, Park SU.

Research Center for Proteineous Materials, Chosun University, Gwangju, Korea.

Buckwheat (Fagopyrum esculentum Moench.) is a potentially important source of
rutin, a natural flavonoid with antihyperglycemic, antihypertensive, and
antioxidative properties. To examine in vitro production of rutin, we established
a hairy root culture of buckwheat by infecting leaf explants with Agrobacterium
rhizogenes R1000, and tested the growth conditions and rutin production rates of
these cultures. Ten hairy root clones were established; their growth and rutin
production rates ranged from 233 to 312 (mg dry wt per 30 mL flask, and 0.8 to
1.2 (mg/g dry wt), respectively. Clone H8, which had high growth and rutin
production rates (312 mg dry wt per 30 mL flask and 1.2 mg/g dry wt,
respectively), was selected for further experiments. H8 showed maximal growth and
rutin content at 30 days in culture in MS medium. Of four tested culture media,
half-strength MS medium was found to induce the highest levels of growth (378 mg
dry wt per 30 mL flask) and rutin production (1.4 mg/g dry wt) by clone H8. In
contrast, supplementation with auxins (0.1-1 mg/l IAA, IBA and NAA) increased the
growth rate, but had no significant effect on rutin production by H8.
Collectively, these findings indicate that hairy root cultures of buckwheat
culture could be a valuable alternative approach for rutin production.

PMID: 17516253 [PubMed - indexed for MEDLINE]

575: Biotechnol Lett. 2007 Jul;29(7):1129-34. Epub 2007 Mar 20.

A cytosolic NADP-malic enzyme gene from rice (Oryza sativa L.) confers salt
tolerance in transgenic Arabidopsis.

Cheng Y, Long M.

Alkali Soil Natural Environmental Science Center, Stress Molecular Biology
Laboratory, Northeast Forestry University, Harbin, PR China.
chengyuxiang111@sina.com.cn

NADP-malic enzyme (NADP-ME, EC 1.1.1.40) functions in many different pathways in
plant and may be involved in plant defense such as wound and UV-B radiation.
Here, expression of the gene encoding cytosolic NADP-ME (cytoNADP-ME, GenBank
Accession No. AY444338) in rice (Oryza sativa L.) seedlings was induced by salt
stress (NaCl). NADP-ME activities in leaves and roots of rice also increased in
response to NaCl. Transgenic Arabidopsis plants over-expressing rice cytoNADP-ME
had a greater salt tolerance at the seedling stage than wild-type plants in MS
medium-supplemented with different levels of NaCl. Cytosolic NADPH/NADP(+)
concentration ratio of transgenic plants was higher than those of wild-type
plants. These results suggest that rice cytoNADP-ME confers salt tolerance in
transgenic Arabidopsis seedlings.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17516134 [PubMed - indexed for MEDLINE]

576: Plant Cell Rep. 2007 Sep;26(9):1555-65. Epub 2007 May 22.

Isolation of the maize Zpu1 gene promoter and its functional analysis in
transgenic tobacco plants.

Chen X, Wang Z, Gu R, Fu J, Wang J, Zhang Y, Wang M, Zhang J, Jia J, Wang G.

State Key Laboratory of Agrobiotechnology and Department of Seed Sciences and
Technology, China Agricultural University, Beijing 100094, China.

By screening a genomic library of maize, a 2.2 kb 5' flanking fragment of Zpu1
gene, encoding the pullulanase-type starch debranching enzyme, was isolated.
Promoter fragments of various lengths, including the full 5' flanking sequence
(-2267 to -1) (Z1), a 3' deletion (-2267 to -513) (Z5) and three 5' deletions
extending to -1943 (Z2), -1143 (Z3) and -516 (Z4) upstream of the translational
initiation codon (ATG), were fused to the GUS reporter gene and introduced into
tobacco. When these constructs were tested in transgenic tobacco plants,
seed-preferred GUS activity was observed in pZ1-transgenic lines. In
pZ2-transgenic lines, the GUS activity was not only restricted to seeds, but was
also detected in calyxes, petals, stamens and mature leaves. At the same time,
negligible GUS activity was detected in roots, stems, young leaves, stigmas and
ovaries from the transgenic tobacco plants, which had integrated the full
isolated sequence of Zpu1 promoter or its deletions. Deletion analysis indicated
that the promoter contained a putative positive cis-regulatory element and the
proximal region (-516 to -1) was essential for directing the expression of gus
reporter gene. Analysis of GUS activity during the fruit development and seed
germination suggested that Zpu1 promoter is active both in starch anabolism and
in starch catabolism, which is consistent with the function of the endogenous
gene in maize. GUS activity in leaves under light and darkness confirmed that
Zpu1 promoter functions in the starch degradation of photosynthetic tissues in
the dark phase of the diurnal cycle.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17516072 [PubMed - indexed for MEDLINE]

577: Plant Cell Rep. 2007 Sep;26(9):1567-73. Epub 2007 May 22.

Agrobacterium-mediated transformation of a low glutelin mutant of 'Koshihikari'
rice variety using the mutated-acetolactate synthase gene derived from rice
genome as a selectable marker.

Wakasa Y, Ozawa K, Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki 305-8602, Japan.
ywakasa@nias.affrc.go.jp

We have developed an efficient rice transformation system that uses only rice
genome-derived components. The transgenic 'Koshihikari' rice, low-glutelin mutant
a123, is capable of accumulating large amounts of bioactive peptides in the
endosperm. Agrobacterium-mediated transformation using the mutated-acetolactate
synthase (mALS) gene expressed under the control of the callus-specific promoter
(CSP) as a selectable marker was used to introduce GFP and an anti-hypertensive
hexapeptide into 'Koshihikari' a123. The CSP:mALS gene cassette confers
pyrimidinyl carboxy herbicide resistance to transgenic rice callus, but is not
expressed in regenerated plants. Transformation efficiency of transgenic rice
line a123 was improved from about 10% to about 30% by modifying callus induction,
callus selection and regeneration media conventionally used for rice tissue
culture.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17516071 [PubMed - indexed for MEDLINE]

578: Plant Mol Biol. 2007 Jul;64(5):613-8. Epub 2007 May 20.

Increased calcium bioavailability in mice fed genetically engineered plants
lacking calcium oxalate.

Morris J, Nakata PA, McConn M, Brock A, Hirschi KD.

Vegetable and Fruit Improvement Center, Texas A&M University, College Station, TX
77845, USA.

Bioavailable calcium affects bone formation and calcification. Here we
investigate how a single gene mutation altering calcium partitioning in the model
forage crop Medicago truncatula affects calcium bioavailability. Previously, the
cod5 M. truncatula mutant was identified which contains identical calcium
concentrations to wild-type, but contains no oxalate crystals. In this study,
equal number of male and female mice were randomly grouped and then fed one of
four 45Ca-containing diets: M. truncatula extrinsically or intrinsically labeled,
and cod5 extrinsically or intrinsically labeled. Absorption of the tracer was
determined in the legs one day after consumption. The absorption was similar in
the M. truncatula and cod5 extrinsically labeled diets; however, in the
intrinsically labeled diets, calcium absorption was 22.87% (P < 0.001) higher in
mice fed cod5. Our study presents the first genetic evidence demonstrating the
nutritional impact of removing oxalate crystals from foods.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17514431 [PubMed - indexed for MEDLINE]

579: Phytochemistry. 2007 Aug-Sep;68(16-18):2290-301. Epub 2007 May 23.

Metabolic flux analysis in plants using dynamic labeling technique: application
to tryptophan biosynthesis in cultured rice cells.

Matsuda F, Wakasa K, Miyagawa H.

Plant Functions and Their Control, CREST, Japan Science and Technology Agency,
3-4-5 Nihonbashi, Chuo, Tokyo 103-0027, Japan.

The concept and methodology of using dynamic labeling for the MFA of plant
metabolic pathways are described, based on a case study to develop a method for
the MFA of the tryptophan biosynthetic pathway in cultured rice cells. Dynamic
labeling traces the change in the labeling level of a metabolite in a metabolic
pathway after the application of a stable isotope-labeled compound. In this
study, [1-(13)C] l-serine was fed as a labeling precursor and the labeling level
of Trp was determined by using the LC-MS/MS. The value of metabolic flux is
determined by fitting a model describing the labeling dynamics of the pathway to
the observed labeling data. The biosynthetic flux of Trp in rice suspension
cultured cell was determined to be 6.0+/-1.1 nmol (gFWh)(-1). It is also
demonstrated that an approximately sixfold increase in the biosynthetic flux of
Trp in transgenic rice cells expressing the feedback-insensitive version of
anthranilate synthase alpha-subunit gene (OASA1D) resulted in a 45-fold increase
in the level of Trp. In this article, the basic workflow for the experiment is
introduced and the details of the actual experimental procedures are explained.
Future perspectives are also discussed by referring recent advances in the
dynamic labeling approach.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17512026 [PubMed - indexed for MEDLINE]

580: Phytochemistry. 2007 Jun;68(11):1497-509. Epub 2007 May 16.

Introduction of sense constructs of cinnamate 4-hydroxylase (CYP73A24) in
transgenic tomato plants shows opposite effects on flux into stem lignin and
fruit flavonoids.

Millar DJ, Long M, Donovan G, Fraser PD, Boudet AM, Danoun S, Bramley PM, Bolwell
GP.

School of Biological Sciences, Royal Holloway, University of London, Egham,
Surrey TW20 0EX, UK.

Understanding regulation of phenolic metabolism underpins attempts to engineer
plants for diverse properties such as increased levels of antioxidant flavonoids
for dietary improvements or reduction of lignin for improvements to fibre
resources for industrial use. Previous attempts to alter phenolic metabolism at
the level of the second enzyme of the pathway, cinnamate 4-hydroxylase have
employed antisense expression of heterologous sequences in tobacco. The present
study describes the consequences of homologous sense expression of tomato
CYP73A24 on the lignin content of stems and the flavonoid content of fruits. An
extensive number of lines were produced and displayed four developmental variants
besides a normal phenotype. These aberrant phenotypes were classified as dwarf
plants, plants with distorted (curly) leaves, plants with long internodes and
plants with thickened waxy leaves. Nevertheless, some of the lines showed the
desired increase in the level of rutin and naringenin in fruit in a normal
phenotype background. However this could not be correlated directly to increased
levels of PAL and C4H expression as other lines showed less accumulation,
although all lines tested showed increases in leaf chlorogenic acid which is
typical of Solanaceous plants when engineered in the phenylpropanoid pathway.
Almost all transgenic lines analysed showed a considerable reduction in stem
lignin and in the lines that were specifically examined, this was correlated with
partial sense suppression of C4H. Although not the primary purpose of the study,
these reductions in lignin were amongst the greatest seen in plants modified for
lignin by manipulation of structural genes. The lignin showed higher syringyl to
coniferyl monomeric content contrary to that previously seen in tobacco
engineered for downregulation of cinnamate 4-hydroxylase. These outcomes are
consistent with placing CYP73A24 more in the lignin pathway and having a role in
flux control, while more complex regulatory processes are likely to be involved
in flavonoid and chlorogenic acid accumulation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17509629 [PubMed - indexed for MEDLINE]

581: J Agric Food Chem. 2007 Jun 13;55(12):4728-34. Epub 2007 May 18.

Development of a certified reference material for genetically modified potato
with altered starch composition.

Broothaerts W, Corbisier P, Emons H, Emteborg H, Linsinger TP, Trapmann S.

European Commission, Joint Research Centre, Institute for Reference Materials and
Measurements (IRMM), Retieseweg 111, 2440 Geel, Belgium.
Wim.Broothaerts@ec.europa.eu

The presence of genetically modified organisms (GMOs) in food and feed products
is subject to regulation in the European Union (EU) and elsewhere. As part of the
EU authorization procedure for GMOs intended for food and feed use, reference
materials must be produced for the quality control of measurements to quantify
the GMOs. Certified reference materials (CRMs) are available for a range of
herbicide- and insect-resistant genetically modified crops such as corn, soybean,
and cotton. Here the development of the first CRM for a GMO that differs from its
non-GMO counterpart in a major compositional constituent, that is, starch, is
described. It is shown that the modification of the starch composition of potato
(Solanum tuberosum L.) tubers, together with other characteristics of the
delivered materials, have important consequences for the certification strategy.
Moreover, the processing and characterization of the EH92-527-1 potato material
required both new and modified procedures, different from those used routinely
for CRMs produced from genetically modified seeds.

PMID: 17508757 [PubMed - indexed for MEDLINE]

582: Plant Cell Rep. 2007 Sep;26(9):1539-46. Epub 2007 May 17.

Combined expression of chitinase and lipid transfer protein genes in transgenic
carrot plants enhances resistance to foliar fungal pathogens.

Jayaraj J, Punja ZK.

Department of Biological Sciences, Simon Fraser University, 8888 University
Drive, Burnaby, BC, Canada V5A 1S6.

Two pathogenesis-related (PR) protein genes consisting of a barley chitinase
(chi-2) and a wheat lipid-transfer-protein (ltp) were introduced singly and in
combination into carrot plants via Agrobacterium-mediated transformation using
the phosphinothricin acetyl transferase (bar) gene as a selectable marker. Over
75% of regenerated plants were confirmed to be positive for the transgenes by PCR
and RT-PCR and were resistant to the herbicide Liberty (0.2%, v/v). Northern
analysis and immunoblotting confirmed the expression of the transgenes in about
70% of the plants, with variable expression levels among individual lines.
Southern analysis revealed from one to three copies of each transgene. Transgenic
plants were inoculated with two necrotrophic foliar fungal pathogens, Alternaria
radicicola and Botrytis cinerea, and showed significantly higher resistance when
both PR genes were expressed compared to single-gene transformants. The level of
disease reduction in plants expressing both genes was 95% for Botrytis and 90%
for Alternaria infection compared to 40-50% for single-gene transformants. The
chi2 and ltp genes could be deployed in combination in other crop plants to
significantly enhance resistance to necrotrophic fungal pathogens.

PMID: 17508215 [PubMed - indexed for MEDLINE]

583: Genes Genet Syst. 2007 Apr;82(2):167-70.

The einkorn wheat (Triticum monococcum) mutant, maintained vegetative phase, is
caused by a deletion in the VRN1 gene.

Shitsukawa N, Ikari C, Shimada S, Kitagawa S, Sakamoto K, Saito H, Ryuto H,
Fukunishi N, Abe T, Takumi S, Nasuda S, Murai K.

Department of Bioscience, Fukui Prefectural University, Japan.

The einkorn wheat (Triticum monococcum) mutant, maintained vegetative phase
(mvp), was induced by nitrogen ion-beam treatment and was identified by its
inability to transit from the vegetative to reproductive phase. In our previous
study, we showed that WAP1 (wheat APETALA1) is a key gene in the regulatory
pathway that controls phase transition from vegetative to reproductive growth in
common wheat. WAP1 is an ortholog of the VRN1 gene that is responsible for
vernalization insensitivity in einkorn wheat. The mvp mutation resulted from
deletion of the VRN1 coding and promoter regions, demonstrating that WAP1/VRN1 is
an indispensable gene for phase transition in wheat. Expression analysis of
flowering-related genes in mvp plants indicated that wheat GIGANTIA (GI),
CONSTANS (CO) and SUPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) genes either
act upstream of or in a different pathway to WAP1/VRN1.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17507783 [PubMed - indexed for MEDLINE]

584: Genetics. 2007 Aug;176(4):2541-9. Epub 2007 May 16.

The in silico map-based cloning of Pi36, a rice coiled-coil nucleotide-binding
site leucine-rich repeat gene that confers race-specific resistance to the blast
fungus.

Liu X, Lin F, Wang L, Pan Q.

Laboratory of Plant Resistasnce and Genetics, College of Resources and
Environmental Sciences, South China Agricultural University, Guangzhou 510642,
China.

The indica rice variety Kasalath carries Pi36, a gene that determines resistance
to Chinese isolates of rice blast and that has been located to a 17-kb interval
on chromosome 8. The genomic sequence of the reference japonica variety
Nipponbare was used for an in silico prediction of the resistance (R) gene
content of the interval and hence for the identification of candidate gene(s) for
Pi36. Three such sequences, which all had both a nucleotide-binding site and a
leucine-rich repeat motif, were present. The three candidate genes were amplified
from the genomic DNA of a number of varieties by long-range PCR, and the
resulting amplicons were inserted into pCAMBIA1300 and/or pYLTAC27 vectors to
determine sequence polymorphisms correlated to the resistance phenotype and to
perform transgenic complementation tests. Constructs containing each candidate
gene were transformed into the blast-susceptible variety Q1063, which allowed the
identification of Pi36-3 as the functional gene, with the other two candidates
being probable pseudogenes. The Pi36-encoded protein is composed of 1056 amino
acids, with a single substitution event (Asp to Ser) at residue 590 associated
with the resistant phenotype. Pi36 is a single-copy gene in rice and is more
closely related to the barley powdery mildew resistance genes Mla1 and Mla6 than
to the rice blast R genes Pita, Pib, Pi9, and Piz-t. An RT-PCR analysis showed
that Pi36 is constitutively expressed in Kasalath.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17507669 [PubMed - indexed for MEDLINE]

585: Mol Plant Microbe Interact. 2007 May;20(5):492-9.

OsWRKY13 mediates rice disease resistance by regulating defense-related genes in
salicylate- and jasmonate-dependent signaling.

Qiu D, Xiao J, Ding X, Xiong M, Cai M, Cao Y, Li X, Xu C, Wang S.

National Key Laboratory of Crop Genetic Improvement, National Center for Plant
Gene Research, Huazhong Agricultural University, Wuhan 430070, China.

Although 109 WRKY genes have been identified in the rice genome, the functions of
most are unknown. Here, we show that OsWRKY13 plays a pivotal role in rice
disease resistance. Overexpression of OsWRKY13 can enhance rice resistance to
bacterial blight and fungal blast, two of the most devastating diseases of rice
worldwide, at both the seedling and adult stages, and shows no influence on the
fertility. This overexpression was accompanied by the activation of salicylic
acid (SA) synthesis-related genes and SA-responsive genes and the suppression of
jasmonic acid (JA) synthesis-related genes and JA-responsive genes. OsWRKY13
bound to the promoters of its own and at least three other genes in SA- and
JA-dependent signaling pathways. Its DNA-binding activity was influenced by
pathogen infection. These results suggest that OsWRKY13, as an activator of the
SA-dependent pathway and a suppressor of JA-dependent pathways, mediates rice
resistance by directly or indirectly regulating the expression of a subset of
genes acting both upstream and downstream of SA and JA. Furthermore, OsWRKY13
will provide a transgenic tool for engineering wider-spectrum and
whole-growth-stage resistance rice in breeding programs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17506327 [PubMed - indexed for MEDLINE]

586: J Agric Food Chem. 2007 Jun 13;55(12):4645-50. Epub 2007 May 16.

Quantification of chlorophyll content and classification of nontransgenic and
transgenic tomato leaves using visible/near-infrared diffuse reflectance
spectroscopy.

Xie L, Ying Y, Ying T.

College of Biosystems Engineering and Food Science, Zhejiang University, 268
Kaixuan Street, 310029 Hangzhou, People's Republic of China.

Visible/near-infrared (vis/NIR) spectroscopy combined with multivariate analysis
was used to quantify chlorophyll content in tomato leaves and classify tomato
leaves with different genes. In this study, transgenic tomato leaves with
antisense LeETR1 (n = 106) and their parent nontransgenic ones (n = 102) were
measured in vis/NIR diffuse reflectance mode. Quantification of chlorophyll
content was achieved by partial least-squares regression with a cross-validation
prediction error equal to 2.87. Partial least-squares discriminant analysis was
performed to classify leaves. The results show that differences between
transgenic and nontransgenic tomato leaves do exist, and excellent classification
can be obtained after optimizing spectral pretreatment. The classification
accuracy can reach to 100% using the derivative of spectral data in the full and
partial wavenumber range. These results demonstrate that vis/NIR spectroscopy
together with chemometrics techniques could be used to quantify chlorophyll
content and differentiate tomato leaves with different genes, which offers the
benefit of avoiding time-consuming, costly, and laborious chemical and sensory
analysis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17503831 [PubMed - indexed for MEDLINE]

587: Proc Natl Acad Sci U S A. 2007 May 22;104(21):8924-9. Epub 2007 May 14.

Construction and behavior of engineered minichromosomes in maize.

Yu W, Han F, Gao Z, Vega JM, Birchler JA.

Division of Biological Sciences, 117 Tucker Hall, University of Missouri,
Columbia, MO 65211, USA.

Engineered minichromosomes were constructed in maize by modifying natural A and
supernumerary B chromosomes. By using telomere-mediated chromosomal truncation,
it was demonstrated that such an approach is feasible for the generation of
minichromosomes of normal A chromosomes by selection of spontaneous polyploid
events that compensate for the deficiencies produced. B chromosomes are readily
fractionated by biolistic transformation of truncating plasmids. Foreign genes
were faithfully expressed from integrations into normal B chromosomes and from
truncated miniB chromosomes. Site-specific recombination between the terminal
transgene on a miniA chromosome and a terminal site on a normal chromosome was
demonstrated. It was also found that the miniA chromosome did not pair with its
progenitor chromosomes during meiosis, indicating a useful property for such
constructs. The miniB chromosomes are faithfully transmitted from one generation
to the next but can be changed in dosage in the presence of normal B chromosomes.
This approach for construction of engineered chromosomes can be easily extended
to other plant species because it does not rely on cloned centromere sequences,
which are species-specific. These platforms will provide avenues for studies on
plant chromosome structure and function and for future developments in
biotechnology and agriculture.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17502617 [PubMed - indexed for MEDLINE]

588: Ecol Lett. 2007 May;10(5):383-93.

Stress and domestication traits increase the relative fitness of crop-wild
hybrids in sunflower.

Mercer KL, Andow DA, Wyse DL, Shaw RG.

Applied Plant Sciences Program, University of Minnesota, 411 Borlaug Hall, 1991
Upper Buford Circle, St Paul, MN 55108, USA. mercer.97@osu.edu

After a decade of transgenic crop production, the dynamics of gene introgression
into wild relatives remain unclear. Taking an ecological genetics approach to
investigating fitness in crop-wild hybrid zones, we uncovered both conditions and
characteristics that may promote introgression. We compared diverse crop-wild
hybrid genotypes relative to wild Helianthus annuus under one benign and three
stressful agricultural environments. Whereas relative fitness of crop-wild
hybrids averaged 0.25 under benign conditions, with herbicide application or
competition it reached 0.45 and was more variable. In some instances, hybrid
fitness matched wild fitness (approximately 1). Thus, wild populations under
agronomic stress may be more susceptible to introgression. Although
'domestication' traits are typically considered unlikely to persist in wild
populations, we found some (e.g. rapid growth and early flowering) that may
enhance hybrid fitness, especially in stressful environments. Rigorous assessment
of how particular genotypes, phenotypes, and environments affect introgression
will improve risk assessment for transgenic crops.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17498137 [PubMed - indexed for MEDLINE]

589: Plant Cell Rep. 2007 Sep;26(9):1681-8. Epub 2007 May 12.

Analysis of a LEA gene promoter via Agrobacterium-mediated transformation of the
desiccation tolerant plant Lindernia brevidens.

Smith-Espinoza C, Bartels D, Phillips J.

Institute of Molecular Physiology and Biotechnology of Plants, University of
Bonn, Kirschallee 1, 53115 Bonn, Germany.

An Agrobacterium tumefaciens-based transformation procedure was developed for the
desiccation tolerant species Lindernia brevidens. Leaf explants were infected
with A. tumefaciens strain GV3101 harbouring a binary vector that carried the
hygromycin resistance gene and an eGFP reporter gene under the control of a
native dehydration responsive LEA promoter (Lb2745pro). PCR analysis of the
selected hygromycin-resistant plants revealed that the transformation rates were
high (14/14) and seeds were obtained from 13/14 of the transgenic lines. A
combination of RNA gel blot and microscopic analyses demonstrated that eGFP
expression was induced upon dehydration and ABA treatment. Comparison with
existing procedures used to transform the well studied resurrection plant and
close relative, Craterostigma plantagineum, revealed that the transformation
process is both rapid and leads to the production of viable seed thus making L.
brevidens a candidate species for functional genomics approaches to determine the
genetic basis of desiccation tolerance.

PMID: 17497152 [PubMed - indexed for MEDLINE]

590: Int Arch Allergy Immunol. 2007;144(1):29-38. Epub 2007 May 11.

A proteomic study to identify soya allergens--the human response to transgenic
versus non-transgenic soya samples.

Batista R, Martins I, Jeno P, Ricardo CP, Oliveira MM.

Instituto Nacional de Saöºde Dr. Ricardo Jorge, Lisboa, Portugal.
rita.batista@insa.min-saude.pt

BACKGROUND: In spite of being among the main foods responsible for allergic
reactions worldwide, soybean (Glycine max)-derived products continue to be
increasingly widespread in a variety of food products due to their
well-documented health benefits. Soybean also continues to be one of the elected
target crops for genetic modification. The aim of this study was to characterize
the soya proteome and, specifically, IgE-reactive proteins as well as to compare
the IgE response in soya-allergic individuals to genetically modified Roundup
Ready soya versus its non-transgenic control. METHODS: We performed
two-dimensional gel electrophoresis of protein extracts from a 5% genetically
modified Roundup Ready flour sample and its non-transgenic control followed by
Western blotting with plasma from 5 soya-sensitive individuals. We used peptide
tandem mass spectrometry to identify soya proteins (55 protein matches),
specifically IgE-binding ones, and to evaluate differences between transgenic and
non-transgenic samples. RESULTS: We identified 2 new potential soybean
allergens--one is maturation associated and seems to be part of the late
embryogenesis abundant proteins group and the other is a cysteine proteinase
inhibitor. None of the individuals tested reacted differentially to the
transgenic versus non-transgenic samples under study. CONCLUSION: Soybean
endogenous allergen expression does not seem to be altered after genetic
modification. Proteomics should be considered a powerful tool for functional
characterization of plants and for food safety assessment. Copyright (c) 2007 S.
Karger AG, Basel.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17496424 [PubMed - indexed for MEDLINE]

591: Acta Biochim Biophys Sin (Shanghai). 2007 May;39(5):377-83.

Inheritance and expression of copies of transgenes 1Dx5 and 1Ax1 in elite wheat
(Triticum aestivum L.) varieties transferred from transgenic wheat through
conventional crossing.

Li S, Wang N, Wang Y, Fang J, He G.

China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory, Huazhong
University of Science and Technology, Wuhan 430074, China.

To study the inheritance and expression of multiple copies of transgenes from
transgenic wheat lines, three crosses between transgenic wheat lines B72-8-11b
and B102-1-2 and Chinese elite wheat varieties Chuan89-107 and Emai18 were
carried out. Chuan89-107x72-8-11b, Chuan89-107x102-1-2 and Emai18x72-8-11b, and
F(1) plants were selfed or backcrossed to obtain different generation
populations. Protein analysis in grains of F(1) and F(2) and backcross progenies
of BC(1)F(1), BC(1)F(2), BC(1)F(3), BC(2)F(1), BC(2)F(2) and BC(2)F(3) by sodium
dodecyl sulfate-polyacrylamide gel electrophoresis showed that the transgenes
1Dx5 and 1Ax1 were expressed and segregated in the target wheat according to
Mendelian laws. A range of 1Dx5 expression levels were observed in the progenies
of Chuan89-107x72-8-11b and Emai18x72-8-11b, but the expression levels of 1Ax1 in
progenies of Chuan89-107x102-1-2 rarely changed. It suggested that the two
foreign genes had different mechanisms of expression in the cross progeny, even
though they were produced in the same way and the foreign 1Dx5 gene of 5-10
copies had the more complicated expression mechanism than the 1Ax1 gene of 4-5
copies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17492135 [PubMed - indexed for MEDLINE]

592: Xenotransplantation. 2007 May;14(3):217-21.

Some ethical issues regarding xenotransfusion.

Roux FA, Saö¯ P, Deschamps JY.

Department of Cellular and Molecular Immuno-Endocrinology, INRA, Nantes School of
Veterinary Medicine, Atlanpole, La Chantrerie, Nantes, France.

BACKGROUND: The use of porcine red blood cells has recently been proposed as a
possible solution to the shortage of blood for human transfusion. OBJECTIVES: The
purpose of this paper is to compare some ethical issues regarding xenotransfusion
(XTF) with those relating to xenotransplantation (XT) of organs, tissues and
cells. MATERIALS AND METHODS: Various ethical concerns and viewpoints relating to
XTF are discussed. RESULTS: The main ethical obstacles to XT do not apply to XTF.
It is much more ethically acceptable to raise pigs for regular blood collection
as it doesn't damage the health of the animal. Porcine endogenous retrovirus
infection, the major concern associated with XT, does not apply to XTF, since red
blood cells have no DNA and have a very short lifespan. Clinical trials will be
possible in humans once XTF has been demonstrated to be effective and harmless in
non-human primates. Transgenesis is acceptable for pig blood donors because only
a limited number of genes are involved, and these animals will never enter into
the livestock gene pool or the food chain. CONCLUSION: Because the need for blood
is less pressing than that for organs, tissues or cells, the use of animal blood
for human transfusion is not an absolute necessity. However, it represents a real
opportunity. The ability to gain access to an unlimited quantity of blood is a
reasonable justification for XTF. Because its technical and ethical hurdles are
less stringent, XTF could be the first large-scale clinical application of XT.

PMID: 17489861 [PubMed - indexed for MEDLINE]

593: J Agric Food Chem. 2007 Jun 13;55(12):4670-7. Epub 2007 May 10.

Recombinant porcine lactoferrin expressed in the milk of transgenic mice enhances
offspring growth performance.

Wu SC, Chen HL, Yen CC, Kuo MF, Yang TS, Wang SR, Weng CN, Chen CM, Cheng WT.

Department of Animal Science and Technology, National Taiwan University, Taipei
106, Taiwan.

The European Commission has proposed a permanent ban on the use of antibiotics as
an ingredient in animal feed to promote growth. Lactoferrin is a globular
multifunctional protein that has been shown to play a role in iron absorption and
to have antimicrobial and anti-inflammatory activities. Therefore, lactoferrin
may serve as a nontherapeutic alternative to antibiotics in livestock husbandry.
As a pilot study toward this goal, transgenic mice have been generated harboring
a porcine lactoferrin (pLF) gene driven by the mammary gland-specific promoter of
the bovine alpha-lactalbumin (alphaLA) gene. The alphaLA-pLF hybrid gene was
confirmed to have been successfully integrated and transmitted stably through the
germ-line in 9 (5 females and 4 males) of 14 transgenic founders. In the female
progenies of six lines analyzed, the transgene copy numbers ranged from 1 to 20
with 1-4 integration sites. Significant levels of pLF protein in milk ranging
from 40 to 106 microg/mL with physical characteristics similar to those of native
pLF in sow's milk were achieved in three of the transgenic lines obtained.
Tissue- and stage-specific pLF expressions were restricted to the mammary gland
of the transgenic female mice during lactation. It was further demonstrated that
the growth performance of animal pups is enhanced by directly feeding the
genetically engineered milk containing enriched pLF protein in transgenic mice.
Furthermore, this enhanced growth performance in suckling mice was proportional
to the concentration of pLF present in milk.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17489602 [PubMed - indexed for MEDLINE]

594: Ecol Appl. 2007 Mar;17(2):431-40.

Air-mediated pollen flow from genetically modified to conventional crops.

Kuparinen A, Schurr F, Tackenberg O, O'Hara RB.

University of Helsinki, Department of Mathematics and Statistics, Finland.
anna.kuparinen@helsinki.fi

Tools for estimating pollen dispersal and the resulting gene flow are necessary
to assess the risk of gene flow from genetically modified (GM) to conventional
fields, and to quantify the effectiveness of measures that may prevent such gene
flow. A mechanistic simulation model is presented and used to simulate pollen
dispersal by wind in different agricultural scenarios over realistic pollination
periods. The relative importance of landscape-related variables such as isolation
distance, topography, spatial configuration of the fields, GM field size and
barrier, and environmental variation are examined in order to find ways to
minimize gene flow and to detect possible risk factors. The simulations
demonstrated a large variation in pollen dispersal and in the predicted amount of
contamination between different pollination periods. This was largely due to
variation in vertical wind. As this variation in wind conditions is difficult to
control through management measures, it should be carefully considered when
estimating the risk of gene flow from GM crops. On average, the predicted level
of gene flow decreased with increasing isolation distance and with increasing
depth of the conventional field, and increased with increasing GM field size.
Therefore, at a national scale and over the long term these landscape properties
should be accounted for when setting regulations for controlling gene flow.
However, at the level of an individual field the level of gene flow may be
dominated by uncontrollable variation. Due to the sensitivity of pollen dispersal
to the wind, we conclude that gene flow cannot be summarized only by the mean
contamination; information about the frequency of extreme events should also be
considered. The modeling approach described in this paper offers a way to predict
and compare pollen dispersal and gene flow in varying environmental conditions,
and to assess the effectiveness of different management measures.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17489250 [PubMed - indexed for MEDLINE]

595: J Agric Food Chem. 2007 May 30;55(11):4281-8. Epub 2007 May 9.

Intragenic crop improvement: combining the benefits of traditional breeding and
genetic engineering.

Rommens CM.

Simplot Plant Sciences, J. R. Simplot Company, Boise, Idaho 83706, USA.
crommens@simplot.com

New crop varieties are developed by applying traditional breeding methods that
rely on random genome modifications. These varieties combine multiple traits that
support farm efficiency and acceptable yields but also contain genes associated
with the production of toxins, allergens, and/or antinutritional compounds that
were not considered during the selection process. Furthermore, existing cultivars
frequently lack the functional genes required for specific sensory traits and the
formation of health-promoting antioxidants. One new method efficiently addresses
some of these issues by either silencing undesirable genes or enhancing the
expression of genes that are linked to dormant beneficial traits. Rather than
incorporating foreign DNA into the plant's genome, these methods transform crops
with plant-derived transfer (P-) DNAs that consist of only native genetic
elements. The genetic modification can be characterized molecularly so that any
inadvertent transfer of undesirable DNA, as may be the case with traditional
methods, is excluded. A recently developed intragenic potato plant is silenced
for the polyphenol oxidase, dikinase R1, and phosphorylase-L genes in a
tuber-specific manner. French fries derived from these tubers lack
discolorations, display an enhanced potato flavor, and produce greatly reduced
amounts of the suspected carcinogen acrylamide. It is argued that intragenic
modification is unlikely to trigger phenotypic, biochemical, or physiological
variation that is new to the species. Similarly, the targeted traits are similar
to those that breeders select for and often have a history of domestication and
reduced fitness. For these reasons, an updated regulatory system is proposed
whereby intragenic crops are considered as low risk and should be cleared for
commercial release in a timely and cost-effective manner. By using modern
techniques to modify the same genetic material that is used by breeders,
intragenic approaches may be perceived as an acceptable extension of traditional
methods in crop improvement.

Publication Types:
Review

PMID: 17488120 [PubMed - indexed for MEDLINE]

596: J Agric Food Chem. 2007 May 30;55(11):4414-21. Epub 2007 May 8.

Sampling plan and test protocol for the semiquantitative detection of genetically
modified canola (Brassica napus) seed in bulk canola seed.

Emslie KR, Whaites L, Griffiths KR, Murby EJ.

National Measurement Institute, Pymble, New South Wales, 2073 Australia.
kerry.emslie@measurement.gov.au

Using a statistical approach, sampling plans for the semiquantitative detection
of genetically modified (GM) canola within a bulk seed sample can be developed
and tailored to meet different GM thresholds, costs, and confidence limits. This
is achieved by changing the number of subsamples analyzed, the number of seeds
per subsample, and the percentage of positive results allowed. These sampling
plans must be devised carefully, taking into account the detection capability of
the analytical assay. This is particularly important in the case of InVigor (a
registered trademark of Bayer CropScience) canola, for which expression levels of
the introduced protein in seed are very low. Lateral flow assays and
enzyme-linked immunosorbent assays (ELISA) were both investigated for their
suitability as a qualitative assay using a subsampling approach. On the basis of
an ELISA, several sampling plans have been devised and validated to provide at
least 99% confidence that bulk seed samples containing at least 0.9% (w/w)
InVigor canola will be detected. Although the term "seed" is used throughout this
paper to refer to the canola, the term "seed" is to be taken to include both seed
and the canola seed (grain) that is harvested by the farmer/grower.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17488033 [PubMed - indexed for MEDLINE]

597: Cell Res. 2007 Aug;17(8):713-21.

OsRRM, a Spen-like rice gene expressed specifically in the endosperm.

Chen SY, Wang ZY, Cai XL.

National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant
Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences, Shanghai, China.

We used the promoter trap technique to identify a rice plant, named 107#, in
which the beta-glucuronidase (GUS) reporter gene was expressed specifically in
the endosperm. A single copy of the T-DNA was inserted into the plant genome, and
a candidate gene OsRRM was identified by the insertion. The OsRRM promoter
directed GUS expression specifically in rice endosperm, analogous to the GUS
expression pattern observed in 107#. OsRRM is a single-copy gene in rice and
encodes a nuclear protein containing 1005 amino-acid residues with two RNA
recognition motifs and one Spen paralog and ortholog C-terminal domain. Western
blot analysis confirmed that the OsRRM protein was specifically expressed in rice
endosperm. Ectopic expression of OsRRM in transgenic plants led to abnormalities,
such as short stature, retarded growth and low fructification rates. Our data, in
conjunction with the reported function of Spen genes, implicated OsRRM in the
regulation of cell development in rice endosperm.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17486123 [PubMed - indexed for MEDLINE]

598: Nat Genet. 2007 Jun;39(6):787-91. Epub 2007 May 7.

Regulation of LANCEOLATE by miR319 is required for compound-leaf development in
tomato.

Ori N, Cohen AR, Etzioni A, Brand A, Yanai O, Shleizer S, Menda N, Amsellem Z,
Efroni I, Pekker I, Alvarez JP, Blum E, Zamir D, Eshed Y.

The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture,
Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel.
ori@agri.huji.ac.il

Plant leaves show pronounced plasticity of size and form. In the classical,
partially dominant mutation Lanceolate (La), the large compound leaves of tomato
(Solanum lycopersicum) are converted into small simple ones. We show that LA
encodes a transcription factor from the TCP family containing an miR319-binding
site. Five independent La isolates are gain-of-function alleles that result from
point mutations within the miR319-binding site and confer partial resistance of
the La transcripts to microRNA (miRNA)-directed inhibition. The reduced
sensitivity to miRNA regulation leads to elevated LA expression in very young La
leaf primordia and to precocious differentiation of leaf margins. In contrast,
downregulation of several LA-like genes using ectopic expression of miR319
resulted in larger leaflets and continuous growth of leaf margins. Our results
imply that regulation of LA by miR319 defines a flexible window of morphogenetic
competence along the developing leaf margin that is required for leaf
elaboration.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17486095 [PubMed - indexed for MEDLINE]

599: Nat Biotechnol. 2007 May;25(5):525-31.

Functional foods from biotech--an unappetizing prospect?

Powell K.

kendall2@nasw.org

In the early 1990s, functional foods promised to solve global malnutrition and
put palatable options for treating ailments on grocery shelves. Since then, a
meager number of products have ripened while the rest have turned sour.

Publication Types:
Review

PMID: 17483833 [PubMed - indexed for MEDLINE]

600: Nat Biotechnol. 2007 May;25(5):509-11.

Compliance costs for regulatory approval of new biotech crops.

Kalaitzandonakes N, Alston JM, Bradford KJ.

Publication Types:
Letter
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17483830 [PubMed - indexed for MEDLINE]

601: Nat Biotechnol. 2007 May;25(5):507-8.

Acceptance of GM food--an experiment in six countries.

Knight JG, Mather DW, Holdsworth DK, Ermen DF.

Publication Types:
Letter
Research Support, Non-U.S. Gov't

PMID: 17483829 [PubMed - indexed for MEDLINE]

602: Nat Biotechnol. 2007 May;25(5):505-6; author reply 506.

Comment on:
Nat Biotechnol. 2006 Dec;24(12):1472-3.

Why the omega-3 piggy should go to market.

Kang JX, Leaf A.

Publication Types:
Comment
Letter

PMID: 17483827 [PubMed - indexed for MEDLINE]

603: Appl Environ Microbiol. 2007 Jul;73(13):4365-7. Epub 2007 May 4.

Mycorrhizal and rhizobial colonization of genetically modified and conventional
soybeans.

Powell JR, Gulden RH, Hart MM, Campbell RG, Levy-Booth DJ, Dunfield KE, Pauls KP,
Swanton CJ, Trevors JT, Klironomos JN.

Department of Integrative Biology, University of Guelph, Guelph, Ontario, Canada.
jpowell@uoguelph.ca

We grew plants of nine soybean varieties, six of which were genetically modified
to express transgenic cp4-epsps, in the presence of Bradyrhizobium japonicum and
arbuscular mycorrhizal fungi. Mycorrhizal colonization and nodule abundance and
mass differed among soybean varieties; however, in no case was variation
significantly associated with the genetic modification.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17483262 [PubMed - indexed for MEDLINE]

604: J Invertebr Pathol. 2007 Jul;95(3):175-80. Epub 2007 Mar 31.

Microbial control and biotechnology research on Bacillus thuringiensis in China.

Huang DF, Zhang J, Song FP, Lang ZH.

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences,
Beijing 100081, China. dfhuang@mail.caas.net.cn

The current status of production and application of biopesticides for pest
control in China is briefly reviewed, with a focus on research advances in
microbial control with Bacillus thuringiensis (Bt). These have led to
improvements in Bt production, exploitation of Bt gene resources, and development
of engineered Bt insecticides and transgenic Bt crops that have expanded host
ranges and increased efficacy against target pests. Both conventional and
biotechnology approaches need to be employed to achieve further progress in
discovery, production technology, formulation processing, development of quality
standards and recommended use patterns.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17481651 [PubMed - indexed for MEDLINE]

605: J AOAC Int. 2007 Mar-Apr;90(2):582-6.

Calculation of measurement uncertainty in quantitative analysis of genetically
modified organisms using intermediate precision--a practical approach.

Zel J, Gruden K, Cankar K, Stebih D, Blejec A.

National Institute of Biology, Vecna pot 111, 1000 Ljubljana, Slovenia.
jana.zel@nib.si

Quantitative characterization of nucleic acids is becoming a frequently used
method in routine analysis of biological samples, one use being the detection of
genetically modified organisms (GMOs). Measurement uncertainty is an important
factor to be considered in these analyses, especially where precise thresholds
are set in regulations. Intermediate precision, defined as a measure between
repeatability and reproducibility, is a parameter describing the real situation
in laboratories dealing with quantitative aspects of molecular biology methods.
In this paper, we describe the top-down approach to calculating measurement
uncertainty, using intermediate precision, in routine GMO testing of food and
feed samples. We illustrate its practicability in defining compliance of results
with regulations. The method described is also applicable to other molecular
methods for a variety of laboratory diagnostics where quantitative
characterization of nucleic acids is needed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17474528 [PubMed - indexed for MEDLINE]

606: Plant Cell Environ. 2007 Jun;30(6):690-9.

RNAi knockdown of Oryza sativa root meander curling gene led to altered root
development and coiling which were mediated by jasmonic acid signalling in rice.

Jiang J, Li J, Xu Y, Han Y, Bai Y, Zhou G, Lou Y, Xu Z, Chong K.

Research Center for Molecular Developmental Biology, Key Laboratory of
Photosynthesis and Environmental Molecular Physiology, Institute of Botany, and
Graduate School of the Chinese Academy of Sciences, Beijing, China.

Jasmonic acid (JA) is a well-known defence hormone, but its biological function
and mechanism in rice root development are less understood. Here, we describe a
JA-induced putative receptor-like protein (OsRLK, AAL87185) functioning in root
development in rice. RNA in situ hybridization revealed that the gene was
expressed largely in roots, and a fusion protein showed its localization on the
plasma membrane. The primary roots in RNAi transgenic rice plants meandered and
curled more easily than wild-type (WT) roots under JA treatment. Thus, this gene
was renamed Oryza sativa root meander curling (OsRMC). The transgenic primary
roots were shorter, the number of adventitious roots increased and the number of
lateral roots decreased as compared to the WT. As well, the second sheath was
reduced in length. Growth of both primary roots and second sheaths was sensitive
to JA treatment. No significant change of JA level appeared in the roots between
the transgenic rice line and WT. Expression of RSOsPR10, involved in the JA
signalling pathway, was induced in transgenic rice. Western blotting revealed
OsRMC induced by JA. Our results suggest that OsRMC of the DUF26 subfamily
involved in JA signal transduction mediates root development and negatively
regulates root curling in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17470145 [PubMed - indexed for MEDLINE]

607: Plant J. 2007 Jun;50(5):917-25. Epub 2007 Apr 23.

Mutated TATA-box/TATA binding protein complementation system for regulated
transgene expression in tobacco.

Chaturvedi CP, Lodhi N, Ansari SA, Tiwari S, Srivastava R, Sawant SV, Tuli R.

National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India.

A two-component expression system was developed to achieve tightly regulated
expression of transgenes in plants. One component functioned as an expression
module whereas the other functioned as a regulatory module. The expression module
comprised a highly expressing TATA-dependent seed-specific promoter in which the
TATA motif in the core promoter was mutated to TGTA. The regulatory module
expressed a mutated general transcription factor TBPm(3) that recognized TGTA and
initiated transcription. Vectors were designed using component one alone or in
combination with component two, and were transformed into tobacco. The TGTA
mutation in the TATA-box completely inactivated the promoter, making component
one non-functional. This non-functional module became transcriptionally active in
the presence of the component two that expressed TBPm(3). The reporter gene gusA
was expressed from the TGTA-containing chimeric legumin promoter, in a tightly
seed-specific manner, in transgenic tobacco plants in the presence of TBPm(3)
that was expressed from a constitutive promoter. The results show that the TGTA
and TBPm(3) combination can be used to achieve high-level tissue-specific
expression of TATA-dependent promoters.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17470060 [PubMed - indexed for MEDLINE]

608: J Anim Sci. 2007 Aug;85(8):1946-52. Epub 2007 Apr 27.

Corn expressing an Escherichia coli-derived phytase gene: a proof-of-concept
nutritional study in pigs.

Nyannor EK, Williams P, Bedford MR, Adeola O.

Department of Animal Science, Purdue University, West Lafayette, IN 47907-2054,
USA.

Two experiments were conducted to investigate the concept that the addition of
corn expressing an Escherichia coli-derived gene (corn-based phytase; CBP) to a
P-deficient diet would improve growth performance and P utilization in pigs. An
E. coli-derived microbial phytase (expressed in Pichia pastoris) sprayed onto a
wheat carrier (Quantum) was included for comparison. In Exp. 1, forty-eight 10-kg
pigs were blocked by BW into 6 blocks and allotted to 8 dietary treatments such
that the BW among dietary treatments was similar and given free access to feed
for 28 d. The dietary treatments were a negative control (NC) with no inorganic P
supplementation; NC + 2, 4, or 6 g of monosodium phosphate/kg; NC + 16,500,
33,000, or 49,500 phytase units (FTU) of CBP/kg; and NC + 16,500 FTU of
Quantum/kg. In Exp. 2, twenty-four 13-kg barrows were assigned to the NC, NC +
16,500 or 33,000 FTU of CBP/kg, or NC + 16,500 FTU of Quantum/kg, in a nutrient-
and energy-balance study consisting of 5 d of adjustment and 5-d collection
periods. The total collection method was used to determine nutrient and energy
balance. Addition of CBP to the low-P NC diet linearly increased (P < 0.01) ADG,
G:F, and plasma P concentration of pigs during the 28-d study. There was no
difference in ADG, G:F, or plasma P concentration between pigs fed the CBP or
Quantum phytase at 16,500 FTU/kg. Weight gain, G:F, and plasma P concentration of
pigs increased (P < 0.01) with monosodium phosphate supplementation, confirming P
deficiency of the NC diet. Linear improvements (P < 0.05) in DM digestibility and
energy retention were observed with CBP supplementation of the NC diet. Although
there were linear (P < 0.01) and quadratic (P < 0.05) increases in N
digestibility, N retention was unaffected by CBP supplementation of the NC diet
in growing pigs. Phosphorus and Ca digestibilities and retentions improved
linearly and quadratically (P < 0.01) with the addition of CBP to the NC diet.
There was no difference in digestive utilization of P or Ca between pigs fed CBP
and Quantum phytase at 16,500 FTU/kg. The data showed that the addition of a corn
expressing an E. coli-derived gene to a P-deficient diet improved growth
performance and indices of P utilization in pigs, and corn expressing phytase was
as efficacious as Quantum phytase when supplemented in P-deficient diets for
weanling pigs.

Publication Types:
Comparative Study

PMID: 17468432 [PubMed - indexed for MEDLINE]

609: Plant Cell. 2007 Apr;19(4):1145-62. Epub 2007 Apr 27.

Maize histone deacetylase hda101 is involved in plant development, gene
transcription, and sequence-specific modulation of histone modification of genes
and repeats.

Rossi V, Locatelli S, Varotto S, Donn G, Pirona R, Henderson DA, Hartings H,
Motto M.

Consiglio per la Ricerca e Sperimentazione in Agricoltura, Istituto Sperimentale
per la Cerealicoltura, Sezione di Bergamo, I-24126 Bergamo, Italy.
vincenzo.rossi@entecra.it

Enzymes catalyzing histone acetylation and deacetylation contribute to the
modulation of chromatin structure, thus playing an important role in regulating
gene and genome activity. We showed that downregulation and overexpression of the
maize (Zea mays) Rpd3-type hda101 histone deacetylase gene induced morphological
and developmental defects. Total levels of acetylated histones and histone
acetylation of both repetitive and nonrepetitive sequences were affected in
hda101 transgenic mutants. However, only transcript levels of genes but not
repeats were altered. In particular, hda101 transgenic mutants showed
differential expression of genes involved in vegetative-to-reproductive
transition, such as liguleless2 and knotted-like genes and their repressor rough
sheath2, which are required for meristem initiation and maintenance. Perturbation
of hda101 expression also affected histone modifications other than acetylation,
including histone H3 dimethylation at Lys-4 and Lys-9 and phosphorylation at
Ser-10. Our results indicate that hda101 affects gene transcription and provide
evidence of its involvement in setting the histone code, thus mediating
developmental programs. Possible functional differences between maize hda101 and
its Arabidopsis thaliana ortholog HDA19 are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17468264 [PubMed - indexed for MEDLINE]

610: Asia Pac J Clin Nutr. 2007;16(2):375-80.

Attitudes of agricultural scientists in Indonesia towards genetically modified
foods.

Februhartanty J, Widyastuti TN, Iswarawanti DN.

SEAMEO-TROPMED, RCCN, University of Indonesia, Campus of UI Salemba, Salemba Raya
no. 6, Jakarta 10430, Indonesia. jfebruhartanty@seameo-rccn.org

Conflicting arguments and partial truths on genetically modified (GM) foods have
left confusion. Although studies of consumer acceptance of GM foods are numerous,
the study of scientists is limited. Therefore, the main objective of this study
was to assess the attitudes of scientists towards GM foods. The study was a cross
sectional study. A total of 400 scientists (involved in at least one of teaching,
research and consultancy) in the Bogor Agricultural Institute, Indonesia were
selected randomly from its faculties of agriculture, veterinary, fishery, animal
husbandry, forestry, agricultural technology, mathematics and science, and the
post graduate department. Data collection was done by face-to-face interview
using a structured questionnaire and self-administered questionnaire. The result
showed that the majority (72.8%) of the respondents were favorably disposed
towards GM foods, 14.8% were neutral, and only 12.5% were against them. The
majority (78.3%) stated that they would try GM food if offered. Most (71%)
reported that they were aware of the term "GM foods". Only half of the
respondents felt that they had a basic understanding about GM foods. However,
based on a knowledge test, 69.8% had a good knowledge score. Nearly 50% indicated
that they were more exposed to news which supported GM foods. Over 90% said that
there should be some form of labeling to distinguish food containing GM
ingredients from non-GM foods. Attitudes were significantly associated with
willingness to try GM foods if offered, restrictions on GM foods, and exposure to
media reports about the pros and cons of GM foods.

PMID: 17468097 [PubMed - indexed for MEDLINE]

611: J Invertebr Pathol. 2007 Jul;95(3):161-7. Epub 2007 Mar 27.

Microbial control in Asia: a bellwether for the future?

Gelernter WD.

PACE Consulting, 1267 Diamond St., San Diego, CA 92109, USA.
gelernter@paceturf.org

Advances and barriers faced by microbial control efforts in Asia offer
instructive insights for microbial control in general. The papers in this series,
which are based on plenary lectures given at the Society for Invertebrate
Pathology 2006 meeting in Wuhan, China, explore the history and current status of
microbial control in China, Japan, and Southeast Asia, and in doing so, bring to
light the following key assumptions that deserve further examination; (1) the
adoption rate of microbial control is well documented; (2) microbial control
agents can compete directly with conventional insecticides; (3) microbial control
agents are relatively easy and inexpensive to produce and develop; (4) patents
will promote innovation and investor interest in microbial control. Alternative
viewpoints are presented that can hopefully aid in future efforts to develop more
safe and effective microbial control agents.

PMID: 17467731 [PubMed - indexed for MEDLINE]

612: Plant Physiol Biochem. 2007 May;45(5):262-9. Epub 2007 Mar 14.

Promoter analysis of iron-deficiency-inducible barley IDS3 gene in Arabidopsis
and tobacco plants.

Kobayashi T, Yoshihara T, Itai RN, Nakanishi H, Takahashi M, Mori S, Nishizawa
NK.

Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1
Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

Under conditions of iron deficiency, graminaceous plants induce the expression of
genes involved in the biosynthesis of mugineic acid family phytosiderophores. We
previously identified the novel cis-acting elements IDE1 and IDE2
(iron-deficiency-responsive element 1 and 2) through promoter analysis of the
barley (Hordeum vulgare L.) iron-deficiency-inducible IDS2 gene in tobacco
(Nicotiana tabacum L.). To gain further insight into plant gene regulation under
iron deficiency, we analyzed the barley iron-deficiency-inducible IDS3 gene,
which encodes mugineic acid synthase. IDS3 promoter fragments were fused to the
beta-glucuronidase (GUS) gene, and this construct was introduced into Arabidopsis
thaliana L. and tobacco plants. In both Arabidopsis and tobacco, GUS activity
driven by the IDS3 promoter showed strongly iron-deficiency-inducible and
root-specific expression. Expression occurred mainly in the epidermis of
Arabidopsis roots, whereas expression was dominant in the pericycle, endodermis,
and cortex of tobacco roots, resembling the expression pattern conferred by IDE1
and IDE2. Deletion analysis revealed that a sequence within -305 nucleotides from
the translation start site was sufficient for specific expression in both
Arabidopsis and tobacco roots. Gain-of-function analysis revealed functional
regions at -305/-169 and -168/-93, whose coexistence was required for the
induction activity in Arabidopsis roots. Multiple IDE-like sequences were
distributed in the IDS3 promoter and were especially abundant within the
functional region at -305/-169. A sequence moderately homologous to that of IDE1
was also present within the -168/-93 region. These IDE-like sequences would be
the first candidates for the functional iron-deficiency-responsive elements in
the IDS3 promoter.

PMID: 17467282 [PubMed - indexed for MEDLINE]

613: J Invertebr Pathol. 2007 Jul;95(3):224-6. Epub 2007 Mar 24.

A global approach to resistance monitoring.

Sivasupramaniam S, Head GP, English L, Li YJ, Vaughn TT.

Monsanto Company, 700 Chesterfield Parkway West, Saint Louis, MO 63017, USA.
sssiva@monsanto.com

Transgenic crops producing insecticidal toxins from the bacterium Bacillus
thuringiensis (Bt) have been grown in many parts of the world since 1996. In the
United States, the Environmental Protection Agency (EPA) has required that
industry submit insect resistance management (IRM) plans for each Bt corn and
cotton product commercialized. A coalition of stakeholders including the EPA,
USDA, academic scientists, industry, and grower organizations have cooperated in
developing specific IRM strategies. Resistance monitoring (requiring submission
of annual reports to the EPA), and a remedial action plan addressing any
contingency if resistance should occur, are important elements of these
strategies. At a global level, Monsanto conducts baseline susceptibility studies
(prior to commercialization), followed by monitoring studies on target pest
populations, for all of its commercialized Bt crop products. To date, Monsanto
has conducted baseline/monitoring studies in Argentina, Australia, Brazil,
Canada, China, Colombia, India, Mexico, the Philippines, South Africa, Spain, and
the United States. Examples of pests on which resistance monitoring has been
conducted include cotton bollworm, Helicoverpa zea, European corn borer, Ostrinia
nubilalis, pink bollworm, Pectinophora gossypiella, Southwestern corn borer,
Diatraea grandiosella, tobacco budworm, Heliothis virescens, and western corn
rootworm, Diabrotica virgifera virgifera, in the United States, cotton bollworm,
Helicoverpa armigera, in China, India and Australia, and H. virescens and H. zea
in Mexico. No field-selected resistance to Bt crops has been documented.

PMID: 17467005 [PubMed - indexed for MEDLINE]

614: Phytochemistry. 2007 Jun;68(11):1521-9. Epub 2007 Apr 26.

Erratum in:
Phytochemistry. 2007 Jul;68(14):2023.

Down-regulation of hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase
in transgenic alfalfa affects lignification, development and forage quality.

Shadle G, Chen F, Srinivasa Reddy MS, Jackson L, Nakashima J, Dixon RA.

Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway,
Ardmore, OK 73401, USA.

The recently discovered enzyme hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl
transferase (HCT) catalyzes the reactions both immediately preceding and
following the insertion of the 3-hydroxyl group into monolignol precursors. A
number of independent transgenic lines of alfalfa (Medicago sativa L.) were
generated in which the levels of HCT were reduced through antisense HCT
expression under control of the bean PAL2 promoter which is preferentially
expressed in vascular tissue. Reduction of enzyme activity in these lines was
from at least 15-50%. The most severely down-regulated lines exhibited
significant stunting, reduction of biomass and delayed flowering. HCT
down-regulation resulted in strongly reduced lignin content and striking changes
in lignin monomer composition, with predominant deposition of 4-hydroxyphenyl
units in the lignin. Vascular structure was impaired in the most strongly
down-regulated lines. Analysis of forage quality parameters showed strong
reductions of neutral- and acid-detergent fiber in the down-regulated lines, in
parallel with large increases (up to 20%) in dry matter forage digestibility.
Although manipulation of lignin biosynthesis can greatly improve forage
digestibility, accompanying effects on plant development need to be better
understood.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17466347 [PubMed - indexed for MEDLINE]

615: Phytochemistry. 2007 Jun;68(11):1545-56. Epub 2007 Apr 26.

Fibrillin influence on plastid ultrastructure and pigment content in tomato
fruit.

Simkin AJ, Gaffö© J, Alcaraz JP, Carde JP, Bramley PM, Fraser PD, Kuntz M.

Laboratoire Plastes et Diffö©renciation Cellulaire, Universitö© Joseph Fourier and
CNRS (UMR5575), BP 53, F-38041 Grenoble cedex 9, France.

The protein termed fibrillin is involved in the formation of lipoprotein
structures, such as plastoglobules and fibrils in certain chromoplast types,
which have been implicated in the over-production of pigments due to a sink
effect. In order to examine its effect in differentiating chromoplasts of a
non-fibrillar type, the pepper fibrillin gene was expressed in tomato fruit. Both
the transcript and protein were found to accumulate during tomato fruit ripening
from an early mature green stage. However, formation of carotenoid deposition
structures in tomato chromoplasts, such as fibrils, was not observed.
Nevertheless, a two-fold increase in carotenoid content and associated carotenoid
derived flavour volatiles (6-methyl-5-hepten-2-one, geranylacetone, beta-ionone
and beta-cyclocitral) was observed. An unexpected phenotypic observation in the
transgenic fruit was the delayed loss of thylakoids in differentiating
chromoplasts, leading to the transient formation of plastids exhibiting a typical
chromoplastic zone adjacent to a protected chloroplastic zone with preserved
thylakoids. An in vitro assay has been developed to monitor fibrillin activity on
thylakoids: data were obtained suggesting a membrane protection role for
fibrillin, more specifically against moderate uncoupling effects.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17466343 [PubMed - indexed for MEDLINE]

616: Plant J. 2007 Jun;50(6):1093-106. Epub 2007 Apr 25.

Deficiency of mitochondrial fumarase activity in tomato plants impairs
photosynthesis via an effect on stomatal function.

Nunes-Nesi A, Carrari F, Gibon Y, Sulpice R, Lytovchenko A, Fisahn J, Graham J,
Ratcliffe RG, Sweetlove LJ, Fernie AR.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie, Am Mö¼hlenberg 1, 14476
Golm, Germany.

Transgenic tomato (Solanum lycopersicum) plants expressing a fragment of a
fumarate hydratase (fumarase) gene in the antisense orientation and exhibiting
considerable reductions in the mitochondrial activity of this enzyme show
impaired photosynthesis. The rate of the tricarboxylic acid cycle was reduced in
the transformants relative to the other major pathways of carbohydrate oxidation
and the plants were characterized by a restricted rate of dark respiration.
However, biochemical analyses revealed relatively little alteration in leaf
metabolism as a consequence of reducing the fumarase activity. That said, in
comparison to wild-type plants, CO(2) assimilation was reduced by up to 50% under
atmospheric conditions and plants were characterized by a reduced biomass on a
whole plant basis. Analysis of further photosynthetic parameters revealed that
there was little difference in pigment content in the transformants but that the
rate of transpiration and stomatal conductance was markedly reduced. Analysis of
the response of the rate of photosynthesis to variation in the concentration of
CO(2) confirmed that this restriction was due to a deficiency in stomatal
function.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17461782 [PubMed - indexed for MEDLINE]

617: J Econ Entomol. 2007 Apr;100(2):573-9.

Evaluation of natural and engineered resistance mechanisms in potato against
Colorado potato beetle in a no-choice field study.

Cooper SG, Douches DS, Coombs JJ, Grafius EJ.

Department of Crop and Soil Sciences, Michigan State University, East Lansing, MI
48824, USA.

The Colorado potato beetle, Leptinotarsa decemlineata Say, is the major insect
pest of potato, Solanum tuberosum L., in eastern North America and is renowned
for resistance development, currently resistant to >40 insecticides worldwide.
Host plant resistance may assist in delaying in resistance development to
insecticides. We evaluated natural host plant resistance mechanisms (glandular
trichomes and Solanum chacoense Bitter-derived resistance) and engineered
resistance mechanisms (Bacillus thuringiensis [Bt] Berliner cry3A and cry1Ia1) in
a no-choice cage study. Six different potato lines representing four host plant
resistance mechanisms were evaluated over 2 yr. Egg masses were placed in each
cage (one egg mass per plant). Almost no feeding was observed in the Bt-cry3A
lines, and only minor feeding was observed in the Bt-cry1Ia1 lines in either
year. On the S. chacoense-derived line, there was significantly less defoliation
than on either the susceptible line or the glandular trichome line in 2003. In
2004, there was significantly higher defoliation on the S. chacoense-derived line
than on the susceptible line or glandular trichome line. The defoliation of the
Solanum chacoense-derived line was largely due to larvae clipping the petioles,
rather than consumption of the leaves. Defoliation on the glandular trichome line
did not differ significantly from the defoliation of the susceptible line,
suggesting glandular trichomes may not be effective in controlling larvae and
preventing defoliation. This study suggested that Bt can provide high levels of
resistance, but the natural resistance mechanisms tested here are variable for
control of Colorado potato beetle larvae in no-choice situations.

Publication Types:
Comparative Study

PMID: 17461085 [PubMed - indexed for MEDLINE]

618: J Econ Entomol. 2007 Apr;100(2):557-65.

Interactions of alternate hosts, postemergence grass control, and
rootworm-resistant transgenic corn on western corn rootworm (Coleoptera:
Chrysomelidae) damage and adult emergence.

Oyediran IO, Higdon ML, Clark TL, Hibbard BE.

Department of Entomology, University of Missouri, Columbia, MO 65211, USA.

Field studies were conducted in 2003 and 2004 to determine the effects of grassy
weeds, postemergence grass control, transgenic rootworm-resistant corn, Zea mays
L., expressing the Cry3Bb1 endotoxin and glyphosate herbicide tolerance (Bt
corn), and the interactions of these factors on western corn rootworm, Diabrotica
virgifera virgifera LeConte, damage and adult emergence. Three insect management
tactics (Bt corn, its nontransgenic isoline, and isoline plus tefluthrin) were
evaluated with two weed species (giant foxtail, Setaria faberi Herrm, and large
crabgrass, Digitaria sanquinalis L. Scop), and four weed management regimes (weed
free, no weed management, early [V3-4] weed management and late [V5-6] weed
management) in a factorial arrangement of a randomized split split-plot design.
In each case, the isoline was also tolerant to glyphosate. Root damage was
significantly affected by insect management tactics in both years, but weed
species and weed management did not significantly affect damage to Bt corn in
either year. Adult emergence was significantly affected by insect management
tactics in both years and by weed species in 2003, but weed management and the
interaction of all three factors was not significant in either year. The sex
ratio of female beetles produced on Bt corn without weeds was generally greater
than when weeds were present and this difference was significant for several
treatments each year. Average dry weight of male and female beetles emerging from
Bt corn was greater than the weights of beetles emerging from isoline or isoline
plus tefluthrin in 2003, but there was no difference for females in 2004 and
males weighed significantly less than other treatments in 2004. The implications
of these results in insect resistance management are discussed.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17461083 [PubMed - indexed for MEDLINE]

619: J Econ Entomol. 2007 Apr;100(2):327-34.

Effects of bacillus thuringiensis transgenic corn on corn earworm and fall
armyworm (Lepidoptera: Noctuidae) densities.

Chilcutt CF, Odvody GN, Correa JC, Remmers J.

Department of Entomology, Texas A&M University Agricultural Research & Extension
Center, Corpus Christi, TX 78406-1412, USA. c-chilcutt@tamu.edu

We examined 17 pairs of near-isogenic hybrids of Bacillus thuringiensis (Bt)
(176, Mon810, and Bt11) and non-Bt corn, Zea mays L., to examine the effects of
Bt on larval densities of Helicoverpa zea (Boddie) and Spodoptera frugiperda (J.
E. Smith) (Lepidoptera: Noctuidae) during 2 yr. During ear formation, instar
densities of H. zea and S. frugiperda were recorded for each hybrid. We found
that H. zea first, second, and fifth instar densities were each affected by
Mon810 and Bt11 Bt corn but not by 176 corn. Surprisingly, first and second
instars were found in higher numbers on ears of Mon810 and Bt11 corn than on
non-Bt corn. Densities of third and fourth instars were equal on Bt and non-Bt
hybrids, whereas densities of fifth instars were lower on Bt plants. S.
frugiperda larval densities were only affected during 1 yr when second, and
fourth to sixth instars were lower on ears of Mon810 and Bt11 hybrids compared
with their non-Bt counterparts. Two likely explanations for early instar H. zea
densities being higher on Bt corn than non-Bt corn are that (1) Bt toxins delay
development, creating a greater abundance of early instars that eventually die,
and (2) reduced survival of H. zea to later instars on Bt corn decreased the
normal asymmetric cannibalism or H. zea-S. frugiperda intraguild predation of
late instars on early instars. Either explanation could explain why differences
between Bt and non-Bt plants were greater for H. zea than S. frugiperda, because
H. zea is more strongly affected by Bt toxins and more cannibalistic.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17461054 [PubMed - indexed for MEDLINE]

620: Clin Vaccine Immunol. 2007 Jun;14(6):685-92. Epub 2007 Apr 25.

Fruit-specific expression of the human immunodeficiency virus type 1 tat gene in
tomato plants and its immunogenic potential in mice.

Ramö­rez YJ, Tasciotti E, Gutierrez-Ortega A, Donayre Torres AJ, Olivera Flores
MT, Giacca M, Gö¨mez Lim MA.

Departamento de Ingenierö­a Genö©tica, Cinvestav Campus Guanajuato, Irapuato, Km
9.6 Libramiento norte, Apartado Postal 629, Irapuato, Gto., Mö©xico 365002.

The human immunodeficiency virus type 1 (HIV-1) Tat protein is considered a
potential candidate vaccine antigen. In an effort to design a strategy for
noninvasive vaccination against HIV-1, we developed transgenic tomatoes
expressing the Tat protein. Two independent plants testing positive in transgene
detection analysis were selected and grown to maturity. Monoclonal antibodies
against Tat recognized a protein of the expected size. Interestingly, expression
of Tat seemed to be toxic to the plant, as in all cases the fruit exhibited
underdeveloped reproductive structures and no seeds. Nine groups of 10
pathogen-free BALB/c male mice were primed either orally, intraperitoneally, or
intramuscularly with 10 mg of tomato fruit extract derived from transgenic or
wild-type plants and with 10 microg of Tat86 recombinant protein. Mice were
immunized at days 0, 14, and 28, and given boosters after 15 weeks; sera were
drawn 7 days after each booster, and the antibody titer was determined by
enzyme-linked immunosorbent assay. All three immunization approaches induced the
development of a strong anti-Tat immunological response, which increased over
time. Isotype subclass determination showed the presence of mucosal
(immunoglobulin A) immunity soon after the beginning of the oral immunization
protocol, and the data were confirmed by the presence of anti-Tat antibodies in
fecal pellets and in vaginal washes. We also demonstrated that sera from
immunized mice inhibited with high efficiency recombinant Tat-dependent
transactivation of the HIV-1 long terminal repeat promoter. This neutralization
activity might be relevant for the suppression of extracellular Tat activities,
which play an important role in HIV disease development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17460112 [PubMed - indexed for MEDLINE]

621: Plant Cell Rep. 2007 Sep;26(9):1635-46. Epub 2007 Apr 26.

Overexpression of SBPase enhances photosynthesis against high temperature stress
in transgenic rice plants.

Feng L, Wang K, Li Y, Tan Y, Kong J, Li H, Li Y, Zhu Y.

Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences,
Wuhan University, Wuhan 430072, People's Republic of China.

Activity of the Calvin cycle enzyme sedoheptulose-1,7-bisphosphatase (SBPase) was
increased by overexpression of a rice plants 9,311 (Oryza sativa L.) cDNA in rice
plants zhonghua11 (Oryza sativa L.). The genetic engineering enabled the plants
to accumulate SBPase in chloroplasts and resulted in enhanced tolerance to high
temperature stress during growth of young seedlings. Moreover, CO(2) assimilation
of transgenic plants was significantly more tolerant to high temperature than
that of wild-type plants. The analyses of chlorophyll fluorescence and the
content and activation of SBPase indicated that the enhancement of photosynthesis
to high temperature was not related to the function of photosystem II but to the
content and activation of SBPase. Western blotting analyses showed that high
temperature stress led to the association of SBPase with the thylakoid membranes
from the stroma fractions. However, such an association was much more pronounced
in wild-type plants than that in transgenic plants. The results in this study
suggested that under high temperature stress, SBPase maintained the activation of
ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) by preventing the
sequestration of Rubisco activase to the thylakoid membranes from the soluble
stroma fraction and thus enhanced the tolerance of CO(2) assimilation to high
temperature stress. The results suggested that overexpression of SBPase might be
an effective method for enhancing high temperature tolerance of plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17458549 [PubMed - indexed for MEDLINE]

622: Crit Rev Food Sci Nutr. 2007;47(4):363-87.

Application of Failure Mode and Effect Analysis (FMEA), cause and effect
analysis, and Pareto diagram in conjunction with HACCP to a corn curl
manufacturing plant.

Varzakas TH, Arvanitoyannis IS.

Technological Educational Institute of Kalamata, School of Agricultural Sciences,
Department of Processing of Agricultural Products, Hellas, Greece.

The Failure Mode and Effect Analysis (FMEA) model has been applied for the risk
assessment of corn curl manufacturing. A tentative approach of FMEA application
to the snacks industry was attempted in an effort to exclude the presence of GMOs
in the final product. This is of crucial importance both from the ethics and the
legislation (Regulations EC 1829/2003; EC 1830/2003; Directive EC 18/2001) point
of view. The Preliminary Hazard Analysis and the Fault Tree Analysis were used to
analyze and predict the occurring failure modes in a food chain system (corn
curls processing plant), based on the functions, characteristics, and/or
interactions of the ingredients or the processes, upon which the system depends.
Critical Control points have been identified and implemented in the cause and
effect diagram (also known as Ishikawa, tree diagram, and the fishbone diagram).
Finally, Pareto diagrams were employed towards the optimization of GMOs detection
potential of FMEA.

Publication Types:
Case Reports
Review

PMID: 17457722 [PubMed - indexed for MEDLINE]

623: Crit Rev Food Sci Nutr. 2007;47(4):335-61.

The politics and science behind GMO acceptance.

Varzakas TH, Arvanitoyannis IS, Baltas H.

T. H. Varzakas Technological Educational Institute of Kalamata, School of
Agricultural Sciences, Department of Processing of Agricultural Products, Hellas,
Greece.

The question of nutritional quality has arisen in the International Community
over the last few years along with other important issues such as population
aging, multipopulation societies, and political conflicts. The nutritional issue
is questioned both quantitatively and qualitatively. It is well known that the
planet faces enormous problems with food that is available. Nowadays 20% of the
population consumes approximately 80% of the produced energy and natural
resources. During the last 15 years, a series of food scares and crises (BSE,
dioxin, foot and mouth disease, bird flu) have seriously undermined public
confidence in food producers and operators and their capacity to produce safe
food. As a result, food safety has become a top priority of the European
legislative authorities. Genetically Modified Organisms (GMOs) is the new food
safety concern which despite the intense reactions from Non Governmental
Organizations and consumer organizations have entered our lives with inadequate
legislative measures to protect consumers from their consumption. The GMO issue
will be the issue for discussion in the long run not only for the European
Community but also for the international community as far as scientific,
economical, political, ideological, ethical, and human issues are concerned.
These issues are discussed in this paper along with a case of study of GM fish.

Publication Types:
Review

PMID: 17457721 [PubMed - indexed for MEDLINE]

624: Plant Cell Rep. 2007 Aug;26(8):1243-51. Epub 2007 Apr 24.

Agrobacterium-mediated transformation of the dwarf pomegranate (Punica granatum
L. var. nana).

Terakami S, Matsuta N, Yamamoto T, Sugaya S, Gemma H, Soejima J.

University of Tsukuba, Tsukuba, Japan.

The dwarf pomegranate (Punica granatum L. var. nana) is a dwarf ornamental plant
that has the potential to be the model plant of perennial fruit trees because it
bears fruits within 1 year of seedling. We established an Agrobacterium-mediated
transformation system for the dwarf pomegranate. Adventitious shoots regenerated
from leaf segments were inoculated with A. tumefaciens strain EHA105 harboring
the binary vector pBin19-sgfp, which contains neomycin phosphotransferase (npt
II) and green fluorescent protein (gfp) gene as a selectable and visual marker,
respectively. After co-cultivation, the inoculated adventitious shoots were cut
into small pieces to induce regeneration, and then selected on MS medium
supplemented with 0.5 muM alpha-naphthaleneacetic acid (NAA), 5 muM
N(6)-benzyladenine (BA), 0.3% gellan gum, 50 mg/l kanamycin, and 10 mg/l
meropenem. Putative transformed shoots were regenerated after 6-8 months of
selection. PCR and PCR-Southern blot analysis revealed the integration of the
transgene into the plant genome. Transformants bloomed and bore fruits within 3
months of being potted, and the inheritance of the transgene was confirmed in
T(1) generations. The advantage of the transformation of dwarf pomegranate was
shown to be the high transformation rate. The establishment of this
transformation system is invaluable for investigating fruit-tree-specific
phenomena.

PMID: 17453216 [PubMed - indexed for MEDLINE]

625: Plant Cell Rep. 2007 Aug;26(8):1275-82. Epub 2007 Apr 24.

Arabidopsis rd29A::DREB1A enhances freezing tolerance in transgenic potato.

Behnam B, Kikuchi A, Celebi-Toprak F, Kasuga M, Yamaguchi-Shinozaki K, Watanabe
KN.

Gene Research Center, Graduate School of Life and Environmental Sciences,
University of Tsukuba, Ten-nodai 1-1-1, Ibaraki, Tsukuba 305-8752, Japan.

The freezing tolerance of 38 independent transgenic potato lines derived from the
cultivar Desiree was tested in vitro using plantlets. The lines were transgenic
for the DREB1A gene under control of the rd29A promoter, both of which were
derived from Arabidopsis thaliana. The level of damage caused by freezing varied
significantly among the transgenic clones and a non-transgenic control (cv.
Desiree). Phenotypic evaluation indicated that the variable responses to freezing
were attributable to genotypic variation, but freezing tolerance was not
dependent on the number of insertions. Northern blot analysis using a DREB1A cDNA
probe revealed high levels of DREB1A expression among the transgenic clones
during the initial cold exposure at 4 degrees C (after 2 h) and in the early
stages of freezing (-20 degrees C, 1-10 min). Furthermore, a linear correlation
was detected between the level of expression and the phenotypic response for all
lines except D138. Thus, in the case of potato, a significant increase in
freezing tolerance was observed in vitro on a small scale following the
introduction of rd29A::DREB1A. Additional testing will show whether this strategy
can be used for tolerance breeding in potato and to increase the freezing
tolerance of other agriculturally important crops.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17453213 [PubMed - indexed for MEDLINE]

626: J Dairy Res. 2007 May;74(2):247-54. Epub 2007 Apr 24.

Lactation induction as a predictor of post-parturition transgene expression in
bovine milk.

Powell A, Kerr D, Guthrie D, Wall R.

Agricultural Research Service, United States Department of Agriculture,
Beltsville, MD, USA.

The bovine's long generation interval results in a delay of several years when
evaluating mammary specific transgenes in genetically engineered animals. This
experiment was conducted to evaluate the feasibility of reducing that waiting
period. Lactation was induced in prepubertal bull and heifer calves as a means of
predicting transgene behaviour during subsequent post-parturient lactations in
the heifers themselves, and in daughters sired by the bulls. The animals carry a
lactation-specific transgene encoding lysostaphin, an antimicrobial protein that
kills Staphlococcus aureus, a mastitis-causing pathogen. Oestrogen, progesterone
and dexamethasone were administered as previously described (Ball et al. 2000) to
nine heifers (five transgenics) ranging in weight from 80 to 145 kg. Eight bull
calves (seven transgenics) weighing 81-178 kg received additional oestrogen and
progesterone injection prior to dexamethasone treatment. All nine heifers
responded to the milk induction scheme yielding between 19 ml and 4.5 l over 5 d.
Milk volume from the four responding males (30 microl to 2.5 ml) was
significantly less than that harvested from females (P=0.025). Only bull calves
>117 kg had a positive response. Lysostaphin was detected in all transgenic
prepubertal heifers and in two transgenic prepubertal bull calves induced. A
positive relationship was observed between lysostaphin's stapholytic activity in
the two types of lactations (r2=0.907, P<0.001) thus providing a useful means of
predicting subsequent lysostaphin production in post-partum milk.

PMID: 17451624 [PubMed - indexed for MEDLINE]

627: Plant Biotechnol J. 2007 Jul;5(4):455-64. Epub 2007 Apr 19.

Resistance to root-knot nematode in tomato roots expressing a nematicidal
Bacillus thuringiensis crystal protein.

Li XQ, Wei JZ, Tan A, Aroian RV.

Section of Cell and Developmental Biology, University of California, San Diego,
La Jolla, CA 92093-0349, USA.

Our laboratory has demonstrated previously that Bacillus thuringiensis (Bt)
crystal (Cry) proteins present in the Cry5 and Cry6 subclades intoxicate
free-living nematodes. In this study, we tested whether the expression of
nematicidal Cry6A in transgenic plants provided protection against
plant-parasitic nematodes. As bacterial codon usage is incompatible with
expression in plants, two different codon-modified cry6A genes were synthesized
for expression in plants. One was designed by maintaining codon diversity whilst
removing codons not common in plants, and the other was designed by selecting the
optimal codon for each amino acid based on the Arabidopsis genome. Both versions
of the cry6A gene, driven by the constitutive cauliflower mosaic virus 35S
promoter, were introduced into tomato roots via Agrobacterium rhizogenes.
Although both were found to express Cry6A protein, the codon diversity gene
generated superior expression. These Cry6A-expressing roots were then challenged
with root-knot nematode, Meloidogyne incognita. Three different infection
parameters were compared between Cry6A-expressing roots and control roots
transformed with empty vector or green fluorescent protein (GFP). These data
demonstrated that M. incognita was able to ingest the 54-kDa Cry6A, and that
Cry6A intoxicated the parasitic nematode, as indicated by a decrease in progeny
production of up to fourfold. These results indicate, for the first time, that a
Bt Cry protein can confer plant resistance to an endoparasitic nematode, and that
Cry proteins have the potential to control plant-parasitic nematodes in
transgenic plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17451491 [PubMed - indexed for MEDLINE]

628: Ig Sanita Pubbl. 2007 Jan-Feb;63(1):65-94.

[Aquaculture in Italy. An integrated model of product quality control]

[Article in Italian]

De Giusti M, Cocchieri RA, De Vito E, Grasso GM, Ortaggi G, Reali D, Ricciardi G,
Romano-Spica V, Boccia A.

Universitö  degli Studi di Roma La Sapienza, Dipartimento de Medicina
Sperimantale, Sezione de Medicina Clinica e Sanitö  Pubblica.

Aquaculture is becoming increasingly diffuse even in Italy. The increased
production introduces new problems such as product quality control and process
safety. This article presents the results of a research project, funded by the
Ministry of the Environment, whose aim was to evaluate and promote aquaculture
product quality and safety in an environmentally responsible way. Four intensive
land-based and offshore aquaculture sites were monitored to evaluate
microbiological, biological and chemical (i.e. polychlorinated biphenyls and
endocrine disruptors) quality of water, products and fish feed. In total 154
samples were analysed, of which 66 were water samples, 55 product samples and 33
feed samples. Salmonella and other enteric pathogens were absent in products and
the aquatic environment, while other environmental pathogens of the Vibrio
species were detected. Bacterial load and fecal indicators were found to be
higher in off-shore products and in mussels from all aquaculture sites. PCBs were
detected in all products in concentrations below 2 microg/g fresh product (Food
and Drug Administration), but on average, higher concentrations were detected in
off-shore products. No estrogen mimetic activity was detected in fish feed, in
contrast it was detected in offshore products and water. Product quality was
found to be strictly correlated with the quality of the environment. Genetically
modified organisms were detected in fish feed but no integration of genetic
material in products occurred.

Publication Types:
English Abstract

PMID: 17450652 [PubMed - indexed for MEDLINE]

629: Plant Physiol. 2007 Jun;144(2):846-56. Epub 2007 Apr 20.

Gene targeting by homologous recombination as a biotechnological tool for rice
functional genomics.

Terada R, Johzuka-Hisatomi Y, Saitoh M, Asao H, Iida S.

National Institute for Basic Biology, Okazaki 444-8585, Japan.

The modification of an endogenous gene into a designed sequence by homologous
recombination, termed gene targeting (GT), has broad implications for basic and
applied research. Rice (Oryza sativa), with a sequenced genome of 389 Mb, is one
of the most important crops and a model plant for cereals, and the single-copy
gene Waxy on chromosome 6 has been modified with a frequency of 1% per surviving
callus by GT using a strong positive-negative selection. Because the strategy is
independent of gene-specific selection or screening, it is in principle
applicable to any gene. However, a gene in the multigene family or a gene
carrying repetitive sequences may preclude efficient homologous
recombination-promoted GT due to the occurrence of ectopic recombination. Here,
we describe an improved GT procedure whereby we obtained nine independent
transformed calli having the alcohol dehydrogenase2 (Adh2) gene modified with a
frequency of approximately 2% per surviving callus and subsequently isolated
eight fertile transgenic plants without the concomitant occurrence of undesirable
ectopic events, even though the rice genome carries four Adh genes, including a
newly characterized Adh3 gene, and a copy of highly repetitive retroelements is
present adjacent to the Adh2 gene. The results indicate that GT using a strong
positive-negative selection can be widely applicable to functional genomics in
rice and presumably in other higher plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17449652 [PubMed - indexed for MEDLINE]

630: Proc Natl Acad Sci U S A. 2007 May 1;104(18):7373-8. Epub 2007 Apr 20.

Comment in:
Proc Natl Acad Sci U S A. 2007 May 1;104(18):7311-2.

Mutational reconstructed ferric chelate reductase confers enhanced tolerance in
rice to iron deficiency in calcareous soil.

Ishimaru Y, Kim S, Tsukamoto T, Oki H, Kobayashi T, Watanabe S, Matsuhashi S,
Takahashi M, Nakanishi H, Mori S, Nishizawa NK.

Department of Global Agricultural Sciences, Graduate School of Agricultural and
Life Sciences, University of Tokyo, 1-1-1 Yayoi, Tokyo 113-8657, Japan.

Iron (Fe) deficiency is a worldwide agricultural problem on calcareous soils with
low-Fe availability due to high soil pH. Rice plants use a well documented
phytosiderophore-based system (Strategy II) to take up Fe from the soil and also
possess a direct Fe2+ transport system. Rice plants are extremely susceptible to
low-Fe supply, however, because of low phytosiderophore secretion and low Fe3+
reduction activity. A yeast Fe3+ chelate-reductase gene refre1/372, selected for
better performance at high pH, was fused to the promoter of the Fe-regulated
transporter, OsIRT1, and introduced into rice plants. The transgene was expressed
in response to a low-Fe nutritional status in roots of transformants. Transgenic
rice plants expressing the refre1/372 gene showed higher Fe3+ chelate-reductase
activity and a higher Fe-uptake rate than vector controls under Fe-deficient
conditions. Consequently, transgenic rice plants exhibited an enhanced tolerance
to low-Fe availability and 7.9x the grain yield of nontransformed plants in
calcareous soils. This report shows that enhancing the Fe3+ chelate-reductase
activity of rice plants that normally have low endogenous levels confers
resistance to Fe deficiency.

PMID: 17449639 [PubMed - indexed for MEDLINE]

631: J Agric Food Chem. 2007 May 16;55(10):3835-42. Epub 2007 Apr 21.

Characterization of protein fractions from Bt-transgenic and non-transgenic maize
varieties using perfusion and monolithic RP-HPLC. Maize differentiation by
multivariate analysis.

Rodrö­guez-Nogales JM, Cifuentes A, Garcö­a MC, Marina ML.

Area de Tecnologö­a de los Alimentos, Departamento de Ing. Agraria y Forestal, ETS
de Ingenierö­as Agrarias, Universidad de Valladolid, 34004 Palencia, Spain.

Protein fractions from transgenic Bt and non-transgenic maize varieties,
extracted by the Osborne solvent fraction procedure, were characterized for the
first time by perfusion and monolithic RP-HPLC in very short analysis times.
Albumins and globulins from different transgenic Bt maizes as well as from their
non-transgenic isogenic varieties were eluted in four peaks using perfusion
RP-HPLC, whereas prolamins and glutelins were separated in seven peaks.
Monolithic RP-HPLC enabled the separation of maize proteins in a large number of
peaks showing 6 and 10 main peaks for albumins and globulins, respectively.
Prolamins migrated at retention times higher than 5 min as seven peaks, whereas
glutelins were separated in three main peaks appearing at retention times higher
than 6.0 min. Moreover, chromatograms of the whole protein extracts showed 8 and
11 components for perfusion and monolithic RP-HPLC, respectively. A comparison of
the chromatograms of the whole protein extracts relative to transgenic and
non-transgenic varieties evidenced quantitative differences on the percentages of
area, mainly for peaks 2 and 3 by perfusion RP-HPLC and for peaks 3 and 7 by
monolithic RP-HPLC. A discriminant analysis based on these proteic profiles was
carried out to classify and predict transgenic Bt maize lines, achieving 100%
correct classification using perfusion RP-HPLC.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17447787 [PubMed - indexed for MEDLINE]

632: Nat Protoc. 2007;2(4):948-52.

Agrobacterium rhizogenes-mediated transformation of soybean to study root
biology.

Kereszt A, Li D, Indrasumunar A, Nguyen CD, Nontachaiyapoom S, Kinkema M,
Gresshoff PM.

ARC Centre of Excellence for Integrative Legume Research, The University of
Queensland, St Lucia, Queensland 4072, Australia.

This protocol is used to induce transgenic roots on soybean to study the function
of genes required in biological processes of the root. Young seedlings with
unfolded cotyledons are infected at the cotyledonary node and/or hypocotyl with
Agrobacterium rhizogenes carrying the gene construct to be tested and the
infection sites are kept in an environment of high humidity. When the emerged
hairy roots can support the plants, the main roots are removed and the transgenic
roots can be tested. Using this method, almost 100% of the infected plants form
hairy roots within 1 month from the start of the experiments.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17446894 [PubMed - indexed for MEDLINE]

633: Science. 2007 Apr 20;316(5823):421-5.

Erratum in:
Science. 2007 Dec 21;318(5858);1866.

Comment in:
Science. 2007 Apr 20;316(5823):377-8.

An evolutionarily conserved mechanism delimiting SHR movement defines a single
layer of endodermis in plants.

Cui H, Levesque MP, Vernoux T, Jung JW, Paquette AJ, Gallagher KL, Wang JY,
Blilou I, Scheres B, Benfey PN.

Department of Biology and Institute for Genome Sciences and Policy, Duke
University, Durham, NC 27708, USA.

Intercellular protein movement plays a critical role in animal and plant
development. SHORTROOT (SHR) is a moving transcription factor essential for
endodermis specification in the Arabidopsis root. Unlike diffusible animal
morphogens, which form a gradient across multiple cell layers, SHR movement is
limited to essentially one cell layer. However, the molecular mechanism is
unknown. We show that SCARECROW (SCR) blocks SHR movement by sequestering it into
the nucleus through protein-protein interaction and a safeguard mechanism that
relies on a SHR/SCR-dependent positive feedback loop for SCR transcription. Our
studies with SHR and SCR homologs from rice suggest that this mechanism is
evolutionarily conserved, providing a plausible explanation why nearly all plants
have a single layer of endodermis.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17446396 [PubMed - indexed for MEDLINE]

634: Science. 2007 Apr 20;316(5823):350-1.

Plant science. Long-sought plant flowering signal unmasked, again.

Pennisi E.

Publication Types:
News

PMID: 17446357 [PubMed - indexed for MEDLINE]

635: Science. 2007 May 18;316(5827):1033-6. Epub 2007 Apr 19.

Hd3a protein is a mobile flowering signal in rice.

Tamaki S, Matsuo S, Wong HL, Yokoi S, Shimamoto K.

Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology,
8916-5 Takayama, Ikoma 630-0101, Japan.

Florigen, the mobile signal that moves from an induced leaf to the shoot apex and
causes flowering, has eluded identification since it was first proposed 70 years
ago. Understanding the nature of the mobile flowering signal would provide a key
insight into the molecular mechanism of floral induction. Recent studies suggest
that the Arabidopsis FLOWERING LOCUS T (FT) gene is a candidate for encoding
florigen. We show that the protein encoded by Hd3a, a rice ortholog of FT, moves
from the leaf to the shoot apical meristem and induces flowering in rice. These
results suggest that the Hd3a protein may be the rice florigen.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17446351 [PubMed - indexed for MEDLINE]

636: J Invertebr Pathol. 2007 Sep;96(1):71-9. Epub 2007 Mar 12.

Efficacy of transgenic rice expressing Cry1Ac and CpTI against the rice
leaffolder, Cnaphalocrocis medinalis (Guenö©e).

Han L, Wu K, Peng Y, Wang F, Guo Y.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of
Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, PR
China.

Detached leaf bioassays, open field tests and cage tests were conducted to
evaluate the control efficacy of two transgenic rice lines, expressing Cry1Ac and
CpTI, against Cnaphalocrocis medinalis (Guenö©e) during 2005-2006 in Fuzhou,
China. Bioassay results showed that cumulative feeding areas of C. medinalis on
transgenic lines were significantly lower than that on control rice lines at
different developmental stages. The corrected mortalities at 96 h after
infestation on transgenic lines during six rice growth stages were >90% and 100%
during experiments conducted in 2005 and 2006, respectively. In the open field
test, there was no significant difference in egg density between transgenic and
control lines during early days of infestation, but significant differences were
detected in late season, due to serious damage on control lines. Larval densities
on control lines were significantly higher than the low larval populations
observed on transgenic lines during both seasons. The percentages of plants with
folded leaves and percentages of folded leaves on transgenic lines were
significantly lower than that on control lines with and without insecticide
applications, during the entire season. In cage tests the cumulative numbers of
C. medinalis adults derived from transgenic lines were significantly lower than
that from control lines with and without insecticide treatments. The high level
of efficacy of the two transgenic rice lines against C. medinalis may provide an
important basis for reduced insecticide applications, an expansion of alternative
pest-control strategies and insect resistance management of Bt rice in the
future.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17445827 [PubMed - indexed for MEDLINE]

637: Environ Biosafety Res. 2006 Jul-Sep;5(3):169-73. Epub 2007 Mar 28.

Gene flow from GM glyphosate-tolerant to conventional soybeans under field
conditions in Japan.

Yoshimura Y, Matsuo K, Yasuda K.

National Institute for Agro-Environmental Sciences, Tsukuba, Ibaraki 305-8604,
Japan. yyoshi@niaes.affrc.go.jp

Natural out-crossing rates were evaluated for conventional soybeans (Glycine max
(L.) Merr.) cultivated adjacent to genetically modified (GM) glyphosate-tolerant
soybeans under field conditions during a four-year period in Japan. A total of
107 846 progeny of 2772 plants harvested from conventional varieties were
screened for glyphosate herbicide tolerance. The highest out-crossing rates,
0.19% in 2001 and 0.16% in 2002, were observed in adjacent rows 0.7 m from the
pollen source. The highest rate in 2004 was 0.052%, which was observed at 2.1 m.
No out-crossing was observed in the rows 10.5 m from the pollen source over the
four-year period. The farthest distances between receptor and pollen source at
which out-crossing was observed were 7 m in 2001, 2.8 m in 2002, and 3.5 m in
2004. The greatest airborne pollen density during the flowering period,
determined by Durham pollen samplers located between the rows of each variety,
was 0.368 grains.cm(-2).day(-1), with the average value at 0.18
grains.cm(-2).day(-1), indicating that the possibility of out-crossing by wind is
minimal. Thrips species and predatory Hemiptera visited the soybean flowers more
frequently during the four-year period than any other common pollinators, such as
bees.

Publication Types:
Comparative Study

PMID: 17445512 [PubMed - indexed for MEDLINE]

638: Environ Biosafety Res. 2006 Jul-Sep;5(3):151-68. Epub 2007 Mar 17.

Meteorological input data requirements to predict cross-pollination of GMO maize
with Lagrangian approaches.

Lipsius K, Wilhelm R, Richter O, Schmalstieg KJ, Schiemann J.

Institute for Geoecology, Environmental Systems Analysis Group, Technical
University Braunschweig, Germany. K.Lipsius@tu-bs.de

Modeling pollen dispersal to predict cross-pollination is of great importance for
the ongoing discussion of adventitious presence of genetically modified material
in food and feed. Two different modeling approaches for pollen dispersal were
used to simulate two years of data for the rate of cross-pollination of non-GM
maize (Zea mays (L.)) fields by pollen from a central 1 ha transgenic field. The
models combine the processes of wind pollen dispersal (transport) and pollen
competition. Both models used for the simulation of pollen dispersal were
Lagrangian approaches: a stochastic particle Lagrange model and a Lagrangian
transfer function model. Both modeling approaches proved to be appropriate for
the simulation of the cross-pollination rates. However, model performance
differed significantly between years. We considered different complexity in
meteorological input data. Predictions compare well with experimental results for
all simplification steps, except that systematic deviations occurred when only
main wind direction was used. Concluding, it can be pointed out that both models
might be adapted to other pollen dispersal experiments of different crops and
plot sizes, when wind direction statistics are available. However, calibration of
certain model parameters is necessary.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17445511 [PubMed - indexed for MEDLINE]

639: Environ Biosafety Res. 2006 Jul-Sep;5(3):127-49. Epub 2007 Mar 24.

The interplay between societal concerns and the regulatory frame on GM crops in
the European Union.

Devos Y, Reheul D, De Waele D, Van Speybroeck L.

Department of Plant Production, Faculty of Bioscience Engineering, Ghent
University, Coupure Links 653, 9000 Ghent, Belgium. Yann.Devos@UGent.be

Recapitulating how genetic modification technology and its agro-food products
aroused strong societal opposition in the European Union, this paper demonstrates
how this opposition contributed to shape the European regulatory frame on GM
crops. More specifically, it describes how this opposition contributed to a de
facto moratorium on the commercialization of new GM crop events in the end of the
nineties. From this period onwards, the regulatory frame has been continuously
revised in order to slow down further erosion of public and market confidence.
Various scientific and technical reforms were made to meet societal concerns
relating to the safety of GM crops. In this context, the precautionary principle,
environmental post-market monitoring and traceability were adopted as ways to
cope with scientific uncertainties. Labeling, traceability, co-existence and
public information were installed in an attempt to meet the general public
request for more information about GM agro-food products, and the specific demand
to respect the consumers' and farmers' freedom of choice. Despite these efforts,
today, the explicit role of public participation and/or ethical consultation
during authorization procedures is at best minimal. Moreover, no legal room was
created to progress to an integral sustainability evaluation during market
procedures. It remains to be seen whether the recent policy shift towards greater
transparency about value judgments, plural viewpoints and scientific
uncertainties will be one step forward in integrating ethical concerns more
explicitly in risk analysis. As such, the regulatory frame stands open for
further interpretation, reflecting in various degrees a continued interplay with
societal concerns relating to GM agro-food products. In this regard, both
societal concerns and diversely interpreted regulatory criteria can be inferred
as signaling a request - and even a quest - to render more explicit the
broader-than-scientific dimension of the actual risk analysis.

PMID: 17445510 [PubMed - indexed for MEDLINE]

640: Environ Biosafety Res. 2006 Jul-Sep;5(3):119-25. Epub 2007 Mar 17.

Problem formulation and hypothesis testing for environmental risk assessments of
genetically modified crops.

Raybould A.

Syngenta, Jealott's Hill International Research Centre, Bracknell, Berkshire RG42
6EY, UK. alan.raybould@syngenta.com

Environmental risk assessments can provide high confidence of minimal risk by
testing theories, "risk hypotheses", that predict the likelihood of unacceptable
harmful events. The creation of risk hypotheses and a plan to test them is called
problem formulation. Effective problem formulation seeks to maximize the
possibility of detecting effects that indicate potential risk; if such effects
are not detected, minimal risk is indicated with high confidence. Two important
implications are that artificial test conditions can increase confidence, whereas
prescriptive data requirements can reduce confidence (increase uncertainty) if
they constrain problem formulation. Poor problem formulation can increase
environmental risk because it leads to the collection of superfluous data that
may delay or prevent the introduction of environmentally beneficial products.

PMID: 17445509 [PubMed - indexed for MEDLINE]

641: J Sep Sci. 2007 Mar;30(4):579-85.

A simple capillary gel electrophoresis approach for efficient and reproducible
DNA separations. Analysis of genetically modified soy and maize.

Sö¡nchez L, Gonzö¡lez R, Crego AL, Cifuentes A.

Institute of Industrial Fermentations (CSIC), Juan de la Cierva 3, Madrid, Spain.

It is generally assumed that in order to achieve suitable separations of DNA
fragments, capillary gel electrophoresis (CGE)-coated capillaries should be used.
In this work, a new method is presented that allows to obtain reproducible CGE
separations of DNA fragments using bare fused-silica capillaries without any
previous coating step. The proposed method only requires: (i) a capillary washing
with 0.1 M hydrochloric acid between injections and (ii) a running buffer
composed of Tris-phosphate-ethylenediamine tetraacetic acid (EDTA) and 4.5% of
2-hydroxyethyl cellulose (HEC) as sieving polymer. The use of this new CGE
procedure gives highly resolved and reproducible separations of DNA fragments
ranging from 50 to 750 bp. The separation of these DNA fragments is accomplished
in less than 30 min with efficiencies up to 1.7 x 10(6) plates/m. Reproducibility
values of migration times (given as %RSD) for the analyzed DNA fragments are
better than 1.0% (n = 4) for the same day, 2.2% (n = 16) for four different days,
and 2.3% (n = 16) for four different capillaries. The usefulness of this
separation method is demonstrated by detecting genetically modified maize and
genetically modified soy after DNA amplification by PCR. This new CGE procedure
together with LIF as detector provides sensitive analysis of 0.9% of Bt11 maize,
Mon810 maize, and Roundup Ready soy in flours with S/ N up to 542. These results
demonstrate the usefulness of this procedure to fulfill the European regulation
on detection of genetically modified organisms in foods.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17444227 [PubMed - indexed for MEDLINE]

642: Dev Neurobiol. 2007 Feb 1;67(2):189-204.

The serotonin receptor SER-1 (5HT2ce) contributes to the regulation of locomotion
in Caenorhabditis elegans.

Dernovici S, Starc T, Dent JA, Ribeiro P.

Institute of Parasitology, McGill University, Macdonald Campus, Ste. Anne de
Bellevue, Quebec, Canada H9X 3V9.

Serotonin (5-hydroxytryptamine: 5HT) is an important neuroactive substance in the
model roundworm, Caenorhabditis elegans. Aside from having effects in feeding and
egg-laying, 5HT inhibits motility and also modulates several locomotory
behaviors, notably food-induced slowing and foraging. Recent evidence showed that
a serotonergic 5HT2-like receptor named SER-1 (also known as 5HT2ce) was
responsible for the effect of 5HT on egg-laying. Here we confirm this observation
and show that SER-1 also plays an important role in locomotion. A mutant lacking
SER-1 was found to be highly resistant to exogenous 5HT in the absence of food
and this resistant phenotype was rescued by reintroducing the SER-1 gene in a
mutant background. Pharmacological studies showed that the same antagonists that
blocked the activity of recombinant SER-1 in vitro also inhibited the effect of
5HT on motility, suggesting the same receptor was responsible for both effects.
When tested for locomotory behaviors, the SER-1 mutant was found to be moderately
defective in food-induced slowing. In addition, the mutant changed direction more
frequently than the wildtype when searching for food, suggesting that SER-1 may
play a role in navigational control during foraging. Both these effects required
the presence of MOD-1, a 5HT gated chloride channel, and the results indicate
that SER-1 and MOD-1 modulate these behaviors through a common pathway. On the
basis of expression analysis of a ser-1::GFP translational fusion, SER-1 is
prominently located in central, integrating neurons of the head ganglia (RIA and
RIC) but not the body wall musculature. The evidence suggests that SER-1 controls
locomotion through indirect modulation of neuromuscular circuits and has effects
both on speed and direction of movement. (c) 2006 Wiley Periodicals, Inc.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17443782 [PubMed - indexed for MEDLINE]

643: Transgenic Res. 2007 Aug;16(4):491-501. Epub 2007 Apr 19.

Transgene flow to hybrid rice and its male-sterile lines.

Jia S, Wang F, Shi L, Yuan Q, Liu W, Liao Y, Li S, Jin W, Peng H.

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences,
Beijing, China. srjia@126.com

Gene flow from genetically modified (GM) crops to the same species or wild
relatives is a major concern in risk assessment. Transgenic rice with insect
and/or disease resistance, herbicide, salt and/or drought tolerance and improved
quality has been successfully developed. However, data on rice gene flow from
environmental risk assessment studies are currently insufficient for the
large-scale commercialization of GM rice. We have provided data on the gene flow
frequency at 17 distances between a GM japonica line containing the bar gene as a
pollen donor and two indica hybrid rice varieties and four male-sterile (ms)
lines. The GM line was planted in a 640 m2 in an isolated experimental plot (2.4
ha), which simulates actual conditions of rice production with pollen
competition. Results showed that: (1) under parallel plantation at the 0-m zone,
the transgene flow frequency to the ms lines ranged from 3.145 to 36.116% and was
significantly higher than that to hybrid rice cultivars (0.037-0.045%). (2) Gene
flow frequency decreased as the distance increased, with a sharp cutoff point at
about 1-2 m; (3) The maximum distance of transgene flow was 30-40 m to rice
cultivars and 40-150 m to ms lines. We believe that these data will be useful for
the risk assessment and management of transgenic rice lines, especially in Asia
where 90% of world's rice is produced and hybrid rice varieties are extensively
used.

PMID: 17443417 [PubMed - indexed for MEDLINE]

644: J Agric Food Chem. 2007 May 16;55(10):4034-42. Epub 2007 Apr 17.

Comparison of the forage and grain composition from insect-protected and
glyphosate-tolerant MON 88017 corn to conventional corn (Zea mays L.).

McCann MC, Trujillo WA, Riordan SG, Sorbet R, Bogdanova NN, Sidhu RS.

Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167,
Covance Laboratories Inc., 3301 Kinsman Boulevard, Madison, Wisconsin 53704, USA.
melinda.c.mccann@monsanto.com

The next generation of biotechnology-derived products with the combined benefit
of herbicide tolerance and insect protection (MON 88017) was developed to
withstand feeding damage caused by the coleopteran pest corn rootworm and
over-the-top applications of glyphosate, the active ingredient in Roundup
herbicides. As a part of a larger safety and characterization assessment, MON
88017 was grown under field conditions at geographically diverse locations within
the United States and Argentina during the 2002 and 2003-2004 field seasons,
respectively, along with a near-isogenic control and other conventional corn
hybrids for compositional assessment. Field trials were conducted using a
randomized complete block design with three replication blocks at each site. Corn
forage samples were harvested at the late dough/early dent stage, ground, and
analyzed for the concentration of proximate constituents, fibers, and minerals.
Samples of mature grain were harvested, ground, and analyzed for the
concentration of proximate constituents, fiber, minerals, amino acids, fatty
acids, vitamins, antinutrients, and secondary metabolites. The results showed
that the forage and grain from MON 88017 are compositionally equivalent to forage
and grain from control and conventional corn hybrids.

Publication Types:
Comparative Study

PMID: 17439144 [PubMed - indexed for MEDLINE]

645: Int J Biometeorol. 2007 Aug;51(6):493-503. Epub 2007 Apr 17.

Gene flow in maize fields with different local pollen densities.

Goggi AS, Lopez-Sanchez H, Caragea P, Westgate M, Arritt R, Clark CA.

Department of Agronomy, 166 Seed Science Center, Iowa State University, Ames, IA
50011, USA. susana@iastate.edu

The development of maize (Zea mays L.) varieties as factories of pharmaceutical
and industrial compounds has renewed interest in controlling pollen dispersal.
The objective of this study was to compare gene flow into maize fields of
different local pollen densities under the same environmental conditions. Two
fields of approximately 36 ha were planted with a nontransgenic, white hybrid, in
Ankeny, Iowa, USA. In the center of both fields, a 1-ha plot of a yellow-seeded
stacked RR/Bt transgenic hybrid was planted as a pollen source. Before flowering,
the white receiver maize of one field was detasseled in a 4:1 ratio to reduce the
local pollen density (RPD). The percentage of outcross in the field with RPD was
42.2%, 6.3%, and 1.3% at 1, 10, and 35 m from the central plot, respectively. The
percentage of outcross in the white maize with normal pollen density (NPD) was
30.1%, 2.7%, and 0.4%, respectively, at these distances. At distances greater
than 100 m, the outcross frequency decreased below 0.1 and 0.03% in the field
with RPD and NPD, respectively. A statistical model was used to compare pollen
dispersal based on observed outcross percentages. The likelihood ratio test
confirmed that the models of outcrossing in the two fields were significantly
different (P is practically 0). Results indicated that when local pollen is low,
the incoming pollen has a competitive advantage and the level of outcross is
significantly greater than when the local pollen is abundant.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17437135 [PubMed - indexed for MEDLINE]

646: Transgenic Res. 2007 Jun;16(3):261-80. Epub 2007 Apr 14.

Biosafety and risk assessment framework for selectable marker genes in transgenic
crop plants: a case of the science not supporting the politics.

Ramessar K, Peremarti A, Gö¨mez-Galera S, Naqvi S, Moralejo M, Muö±oz P, Capell T,
Christou P.

Departament de Produccio Vegetal i Ciencia Forestal, Universitat de Lleida, Av.
Alcalde Rovira Roure, 191, Lleida 25198, Spain.

Selectable marker gene systems are vital for the development of transgenic crops.
Since the creation of the first transgenic plants in the early 1980s and their
subsequent commercialization worldwide over almost an entire decade, antibiotic
and herbicide resistance selectable marker gene systems have been an integral
feature of plant genetic modification. Without them, creating transgenic crops is
not feasible on purely economic and practical terms. These systems allow the
relatively straightforward identification and selection of plants that have
stably incorporated not only the marker genes but also genes of interest, for
example herbicide tolerance and pest resistance. Bacterial antibiotic resistance
genes are also crucial in molecular biology manipulations in the laboratory. An
unprecedented debate has accompanied the development and commercialization of
transgenic crops. Divergent policies and their implementation in the European
Union on one hand and the rest of the world on the other (industrialized and
developing countries alike), have resulted in disputes with serious consequences
on agricultural policy, world trade and food security. A lot of research effort
has been directed towards the development of marker-free transformation or
systems to remove selectable markers. Such research has been in a large part
motivated by perceived problems with antibiotic resistance selectable markers;
however, it is not justified from a safety point of view. The aim of this review
is to discuss in some detail the currently available scientific evidence that
overwhelmingly argues for the safety of these marker gene systems. Our
conclusion, supported by numerous studies, most of which are commissioned by some
of the very parties that have taken a position against the use of antibiotic
selectable marker gene systems, is that there is no scientific basis to argue
against the use and presence of selectable marker genes as a class in transgenic
plants.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17436060 [PubMed - indexed for MEDLINE]

647: Plant Physiol. 2007 Jun;144(2):890-903. Epub 2007 Apr 13.

Genetic dissection of histidine biosynthesis in Arabidopsis.

Muralla R, Sweeney C, Stepansky A, Leustek T, Meinke D.

Department of Botany, Oklahoma State University, Stillwater, Oklahoma 74078, USA.

The biosynthesis of histidine (His) in microorganisms, long studied through the
isolation and characterization of auxotrophic mutants, has emerged as a paradigm
for the regulation of metabolism and gene expression. Much less is known about
His biosynthesis in flowering plants. One limiting factor has been the absence of
large collections of informative auxotrophs. We describe here the results of a
systematic screen for His auxotrophs of Arabidopsis (Arabidopsis thaliana). Ten
insertion mutants disrupted in four different biosynthetic genes (HISN2, HISN3,
HISN4, HISN6A) were identified through a combination of forward and reverse
genetics and were shown to exhibit an embryo-defective phenotype that could be
rescued by watering heterozygous plants with His. Male transmission of the mutant
allele was in several cases reduced. Knockouts of two redundant genes (HISN1B and
HISN5A) had no visible phenotype. Another mutant blocked in the final step of His
biosynthesis (hisn8) and a double mutant altered in the redundant first step of
the pathway (hisn1a hisn1b) exhibited a combination of gametophytic and embryonic
lethality in heterozygotes. Homozygous mutant seedlings and callus tissue
produced from rescued seeds appeared normal when grown in the presence of His but
typically senesced after continued growth in the absence of His. These knockout
mutants document the importance of His biosynthesis for plant growth and
development, provide valuable insights into amino acid transport and source-sink
relationships during seed development, and represent a significant addition to
the limited collection of well-characterized auxotrophs in flowering plants.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17434988 [PubMed - indexed for MEDLINE]

648: Curr Opin Plant Biol. 2007 Jun;10(3):236-44. Epub 2007 Apr 16.

Engineering oilseeds for sustainable production of industrial and nutritional
feedstocks: solving bottlenecks in fatty acid flux.

Cahoon EB, Shockey JM, Dietrich CR, Gidda SK, Mullen RT, Dyer JM.

US Department of Agriculture-Agricultural Research Service Plant Genetics
Research Unit, Donald Danforth Plant Science Center, 975 North Warson Road, Saint
Louis, Missouri 63132, USA. ecahoon@danforthcenter.org

Oilseeds provide a unique platform for the production of high-value fatty acids
that can replace non-sustainable petroleum and oceanic sources of specialty
chemicals and aquaculture feed. However, recent efforts to engineer the seeds of
crop and model plant species to produce new types of fatty acids, including
hydroxy and conjugated fatty acids for industrial uses and long-chain omega-3
polyunsaturated fatty acids for farmed fish feed, have met with only modest
success. The collective results from these studies point to metabolic
'bottlenecks' in the engineered plant seeds that substantially limit the
efficient or selective flux of unusual fatty acids between different substrate
pools and ultimately into storage triacylglycerol. Evidence is emerging that
diacylglycerol acyltransferase 2, which catalyzes the final step in
triacylglycerol assembly, is an important contributor to the synthesis of unusual
fatty acid-containing oils, and is likely to be a key target for future oilseed
metabolic engineering efforts.

Publication Types:
Review

PMID: 17434788 [PubMed - indexed for MEDLINE]

649: Biochim Biophys Acta. 2007 Apr;1769(4):220-7. Epub 2007 Mar 12.

A novel rice C2H2-type zinc finger protein lacking DLN-box/EAR-motif plays a role
in salt tolerance.

Huang J, Yang X, Wang MM, Tang HJ, Ding LY, Shen Y, Zhang HS.

State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing
Agricultural University, Nanjing 210095, China.

A cDNA for the gene ZFP182, encoding a C2H2-type zinc finger protein, was cloned
from rice by RT-PCR. ZFP182 codes an 18.2 kDa protein with two C2H2-type zinc
finger motifs, one nuclear localization signal and one Leu-rich domain. The
DLN-box/EAR-motif, which exists in most of plant C2H2-type zinc finger proteins,
does not exist in ZFP182. The expression analysis showed that ZFP182 gene was
constitutively expressed in leaves, culms, roots and spikes at the adult rice
plants, and markedly induced in the seedlings by cold (4 degrees C), 150 mM NaCl
and 0.1 mM ABA treatments. The approximate 1.4 kb promoter region of ZFP182 gene
was fused into GUS reporter gene and transformed into tobacco. The histochemical
analysis revealed that GUS expression could not be detected in transformed
tobacco seedlings under normal conditions, but strongly observed in tobacco leaf
discs and the vascular tissue of roots treated with NaCl or KCl. Expression of
ZFP182 in transgenic tobacco and overexpression in rice increased plant tolerance
to salt stress. These results demonstrated that ZFP182 might be involved in plant
responses to salt stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17434609 [PubMed - indexed for MEDLINE]

650: Trends Biotechnol. 2007 Jun;25(6):239-41. Epub 2007 Apr 12.

Reduced terpene levels in cottonseed add food to fiber.

Townsend BJ, Llewellyn DJ.

CSIRO Plant Industry, GPO Box 1600, Canberra, ACT 2601, Australia.
belinda.townsend@bbsrc.ac.uk

Using RNA interference (RNAi) technology, the levels of a toxic phytoprotectant
have recently been reduced specifically in the seeds of cotton to generate a
novel dual-purpose crop. By engineering an endogenous terpene pathway, there is
now the exciting potential for an added-value, genetically modified crop with the
cash value of the fiber supported by the improved nutritional value and expanded
food and feed use for the cottonseed, which is normally a low-value by-product.

Publication Types:
Review

PMID: 17433845 [PubMed - indexed for MEDLINE]

651: Plant Cell Rep. 2007 Aug;26(8):1133-54. Epub 2007 Apr 13.

Molecular genetic improvement of cereals: transgenic wheat (Triticum aestivum
L.).

Vasil IK.

University of Florida, Gainesville, FL 32611-0690, USA. ivasil@ufl.edu

Only modest progress has been made in the molecular genetic improvement of wheat
following the production of the first transgenic plants in 1992, made possible by
the development of efficient, long-term regenerable embryogenic cultures derived
from immature embryos and use of the biolistics method for the direct delivery of
DNA into regenerable cells. Transgenic lines expressing genes that confer
resistance to environmentally friendly non-selective herbicides, and pests and
pathogens have been produced, in addition to lines with improved bread-making and
nutritional qualities; some of these are ready for commercial production.
Reduction of losses caused by weeds, pests and pathogens in such plants not only
indirectly increases available arable land and fresh water supplies, but also
conserves energy and natural resources. Nevertheless, the work carried out thus
far can be considered only the beginning, as many difficult tasks lie ahead and
much remains to be done. The challenge now is to produce higher-yielding
varieties that are more nutritious, and are resistant or tolerant to a wide
variety of biotic as well as abiotic stresses (especially drought, salinity,
heavy metal toxicity) that currently cause substantial losses in productivity.
How well we will meet this challenge for wheat, and indeed for other cereal and
non-cereal crops, will depend largely on establishing collaborative partnerships
between breeders, molecular biologists, biotechnologists and industry, and on how
effectively they make use of the knowledge and insights gained from basic studies
in plant biology and genetics, the sequencing of plant/cereal genomes, the
discovery of synteny in cereals, and the availability of DNA-based markers and
increasingly detailed chromosomal maps.

Publication Types:
Review

PMID: 17431631 [PubMed - indexed for MEDLINE]

652: Environ Health Perspect. 2007 Mar;115(3):354-60. Epub 2006 Dec 19.

Low-dose exposure and immunogenicity of transgenic maize expressing the
Escherichia coli heat-labile toxin B subunit.

Beyer AJ, Wang K, Umble AN, Wolt JD, Cunnick JE.

Interdepartmental Microbiology, Iowa State University, Ames, Iowa 50011, USA.

BACKGROUND: Transgenic maize, which produces the nontoxic B subunit of the
Escherichia coli heat-labile toxin (LT-B) in seed, has proven to be an effective
oral immunogen in mice. Currently, there is considerable concern over accidental
consumption of transgenic maize expressing LT-B by humans and domestic animals.
We have yet to define nonimmunogenic levels of transgenic LT-B when ingested.
OBJECTIVES: Our goal in this study was to determine the highest dose of LT-B
orally administered in mice that does not result in a measurable immune response.
We defined an immune response as specific serum or mucosal IgG or IgA
significantly greater than background after three feedings (0.0002-20 mug) or a
priming response induced by the intermittent feeding. METHODS: We fed transgenic
maize pellets on days 0, 7, 21, and 49 and collected serum and fecal samples
weekly. Serum was analyzed for LT-B-specific IgG and IgA, and feces was analyzed
for LT-B-specific IgA. RESULTS: We observed a dose-dependent anti-LT-B antibody
response with high specific antibody concentrations in groups fed high doses
(0.2, 2, 20 mug) of LT-B maize. Mice fed 0.02 mug LT-B demonstrated immune
priming in 62.5% of the animals. Mice that were fed increase in specific antibody nor did they demonstrate immune priming, indicating
that 0.002 mug LT-B was the highest nonimmunogenic dose tested. CONCLUSION: Our
results demonstrate that LT-B derived from transgenic maize is immunogenic at
nanogram levels when orally administered to mice.

PMID: 17431483 [PubMed - indexed for MEDLINE]

653: J Exp Bot. 2007;58(7):1603-15. Epub 2007 Apr 12.

Calystegines in potatoes with genetically engineered carbohydrate metabolism.

Richter U, Sonnewald U, Drö¤ger B.

Department of Pharmacy, Faculty of Science I Biosciences, Martin Luther
University Halle-Wittenberg, Hoher Weg 8, D-06120 Halle, Germany.

Calystegines are hydroxylated nortropane alkaloids derived from the tropane
alkaloid biosynthetic pathway. They are strong glycosidase inhibitors and occur
in vegetables such as potatoes, tomatoes, and cabbage. Calystegine accumulation
in root cultures was described to increase with carbohydrate availability.
Whether this is indicative for the in planta situation is as yet unknown.
Potatoes are model plants for the study of carbohydrate metabolism. Numerous
transgenic potato lines with altered carbohydrate metabolism are available, but
rarely were examined for alterations in secondary metabolism. In this study,
calystegine accumulation and expression of biosynthetic enzymes were related to
genetic modifications in carbohydrate metabolism in potato tubers. Tubers
contained more soluble sugars due to overexpression of yeast invertase in the
apoplast or in the cytosol, or due to antisense suppression of sucrose synthase.
It is shown that the major part of calystegines in tubers originated from
biosynthesis in plant roots. Yet, tuber calystegine levels responded to genetic
alterations of carbohydrate metabolism in tubers. The strongest increase in
calystegines was found in tubers with suppressed sucrose synthase activity.
Transcripts and enzyme activities involved in calystegine biosynthesis largely
concurred with product accumulation. Whole plant organs were examined similarly
and displayed higher calystegines and corresponding enzyme activities in roots
and stolons of plants with enhanced soluble sugars. Increases in calystegines
appear to be linked to sucrose availability.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17431026 [PubMed - indexed for MEDLINE]

654: Plant Biotechnol J. 2007 May;5(3):431-41.

Increasing seed oil content in oil-seed rape (Brassica napus L.) by
over-expression of a yeast glycerol-3-phosphate dehydrogenase under the control
of a seed-specific promoter.

Vigeolas H, Waldeck P, Zank T, Geigenberger P.

Max-Planck Institute of Molecular Plant Physiology, Am Mö¼hlenberg 1, 14476 Golm,
Germany.

Previous attempts to manipulate oil synthesis in plants have mainly concentrated
on the genes involved in the biosynthesis and use of fatty acids, neglecting the
possible role of glycerol-3-phosphate supply on the rate of triacylglycerol
synthesis. In this study, a yeast gene coding for cytosolic glycerol-3-phosphate
dehydrogenase (gpd1) was expressed in transgenic oil-seed rape under the control
of the seed-specific napin promoter. It was found that a twofold increase in
glycerol-3-phosphate dehydrogenase activity led to a three- to fourfold increase
in the level of glycerol-3-phosphate in developing seeds, resulting in a 40%
increase in the final lipid content of the seed, with the protein content
remaining substantially unchanged. This was accompanied by a decrease in the
glycolytic intermediate dihydroxyacetone phosphate, the direct precursor of
glycerol-3-phosphate dehydrogenase. The levels of sucrose and various metabolites
in the pathway from sucrose to fatty acids remained unaltered. The results show
that glycerol-3-phosphate supply co-limits oil accumulation in developing seeds.
This has important implications for strategies that aim to increase the overall
level of oil in commercial oil-seed crops for use as a renewable alternative to
petrol.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17430545 [PubMed - indexed for MEDLINE]

655: Plant Biotechnol J. 2007 May;5(3):361-80.

Emerging trends in the functional genomics of the abiotic stress response in crop
plants.

Vij S, Tyagi AK.

Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular
Biology, University of Delhi South Campus, New Delhi, India.

Plants are exposed to different abiotic stresses, such as water deficit, high
temperature, salinity, cold, heavy metals and mechanical wounding, under field
conditions. It is estimated that such stress conditions can potentially reduce
the yield of crop plants by more than 50%. Investigations of the physiological,
biochemical and molecular aspects of stress tolerance have been conducted to
unravel the intrinsic mechanisms developed during evolution to mitigate against
stress by plants. Before the advent of the genomics era, researchers primarily
used a gene-by-gene approach to decipher the function of the genes involved in
the abiotic stress response. However, abiotic stress tolerance is a complex trait
and, although large numbers of genes have been identified to be involved in the
abiotic stress response, there remain large gaps in our understanding of the
trait. The availability of the genome sequences of certain important plant
species has enabled the use of strategies, such as genome-wide expression
profiling, to identify the genes associated with the stress response, followed by
the verification of gene function by the analysis of mutants and transgenics.
Certain components of both abscisic acid-dependent and -independent cascades
involved in the stress response have already been identified. Information
originating from the genome-wide analysis of abiotic stress tolerance will help
to provide an insight into the stress-responsive network(s), and may allow the
modification of this network to reduce the loss caused by stress and to increase
agricultural productivity.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17430544 [PubMed - indexed for MEDLINE]

656: Plant Mol Biol. 2007 Jun;64(3):329-47. Epub 2007 Apr 11.

A barley activation tagging system.

Ayliffe MA, Pallotta M, Langridge P, Pryor AJ.

CSIRO Plant Industry, Box 1600, Clunies Ross Drive, Canberra, ACT, 2601,
Australia. michael.ayliffe@csiro.au

Activation tagging, as the result of random genomic insertion of either promoter
or enhancer sequences, can produce novel, dominant mutations by over-expression
of endogenous genes. This powerful genomics tool has been used extensively in
dicot species such as Arabidopsis, while rice is the only cereal for which an
equivalent system exists. In this study we describe an activation tagging system
in barley based upon the maize Ac/Ds transposable element system. A modified Ds
element (UbiDs) containing two maize polyubiquitin promoters, transposed in
families derived from multiple independent UbiDs transformants and generated new
Ds insertion events at frequencies ranging from 0% to 52% per family. The
majority of transposed UbiDs elements activated high levels of adjacent flanking
sequence transcription. Transposon-mediated expression was detected in all barley
cell and tissue types analysed suggesting that this system is applicable to all
aspects of plant development and biogenesis. In addition to transcriptional
activation, this system is also capable of generating insertional knockout
mutants and a UbiDs inactivated allele of the granule bound starch synthase I
gene (waxy) was recovered that lead to reduced amylose accumulation. The recovery
and analysis of dominant over-expression phenotypes generated by this system will
provide a novel approach to understanding gene function in large cereal genomes
where gene redundancy may mask conventional loss-of-function mutations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17429742 [PubMed - indexed for MEDLINE]

657: Proc Natl Acad Sci U S A. 2007 Apr 17;104(16):6864-9. Epub 2007 Apr 11.

Smallpox subunit vaccine produced in Planta confers protection in mice.

Golovkin M, Spitsin S, Andrianov V, Smirnov Y, Xiao Y, Pogrebnyak N, Markley K,
Brodzik R, Gleba Y, Isaacs SN, Koprowski H.

Biotechnology Foundation Laboratories, Thomas Jefferson University, Philadelphia,
PA 19107-6799, USA.

We report here the in planta production of the recombinant vaccinia virus B5
antigenic domain (pB5), an attractive component of a subunit vaccine against
smallpox. The antigenic domain was expressed by using efficient transient and
constitutive plant expression systems and tested by various immunization routes
in two animal models. Whereas oral administration in mice or the minipig with
collard-derived insoluble pB5 did not generate an anti-B5 immune response,
intranasal administration of soluble pB5 led to a rise of B5-specific
immunoglobulins, and parenteral immunization led to a strong anti-B5 immune
response in both mice and the minipig. Mice immunized i.m. with pB5 generated an
antibody response that reduced virus spread in vitro and conferred protection
from challenge with a lethal dose of vaccinia virus. These results indicate the
feasibility of producing safe and inexpensive subunit vaccines by using plant
production systems.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17428917 [PubMed - indexed for MEDLINE]

658: Tsitol Genet. 2007 Jan-Feb;41(1):3-8.

[Phosphinothricin-resistant somatic hybrids Brassica napus + Orychophragmus
violaceus]

[Article in Ukrainian]

Sakhno LO, Komarnyts'kyä­ IK, Cherep MN, Kuchuk MV.

Phosphinothricin (PPT) resistant hybrid plants between Brassica napus L. cv.
Kalinovsky and Orychophragmus violaceus (L.) O.E. Shulz. were obtained as a
result of somatic hybridization experiments. The hybrids inherited PPT resistance
from O. violaceus plants which were previously transformed by the vector
containing Spm/dSpm Zea mays transposon system with bar gene located within the
nonautonomous transposon. The obtained plants had intermediate morphology. Their
hybrid nature has been confirmed by isozyme (esterase and amilase activity) and
PCR (bar, gus, Spm/dSpm integration) analyses. The hybrids combined B. napus
plastom and O. violaceus mithochondrion that was revealed by PCR-RFLP. The hybrid
plants might be included to rapeseed breeding programme after examination of
their oil quality as well as to chloroplast transformation experiments that is
still urgent for B. napus.

Publication Types:
English Abstract

PMID: 17427411 [PubMed - indexed for MEDLINE]

659: Theor Appl Genet. 2007 Jun;115(1):35-46. Epub 2007 Apr 11.

Over-expression of a LEA gene in rice improves drought resistance under the field
conditions.

Xiao B, Huang Y, Tang N, Xiong L.

National Center of Plant Gene Research (Wuhan), National Key Laboratory of Crop
Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China.

Late embryogenesis abundant (LEA) proteins have been implicated in many stress
responses of plants. In this report, a LEA protein gene OsLEA3-1 was identified
and over-expressed in rice to test the drought resistance of transgenic lines
under the field conditions. OsLEA3-1 is induced by drought, salt and abscisic
acid (ABA), but not by cold stress. The promoter of OsLEA3-1 isolated from the
upland rice IRAT109 exhibits strong activity under drought- and salt-stress
conditions. Three expression constructs consisting of the full-length cDNA driven
by the drought-inducible promoter of OsLEA3-1 (OsLEA3-H), the CaMV 35S promoter
(OsLEA3-S), and the rice Actin1 promoter (OsLEA3-A) were transformed into the
drought-sensitive japonica rice Zhonghua 11. Drought resistance pre-screening of
T(1) families at anthesis stage revealed that the over-expressing families with
OsLEA3-S and OsLEA3-H constructs had significantly higher relative yield (yield
under drought stress treatment/yield under normal growth conditions) than the
wild type under drought stress conditions, although a yield penalty existed in
T(1) families under normal growth conditions. Nine homozygous families,
exhibiting over-expression of a single-copy of the transgene and relatively low
yield penalty in the T(1) generation, were tested in the field for drought
resistance in the T(2) and T(3) generations and in the PVC pipes for drought
tolerance in the T(2) generation. Except for two families (transformed with
OsLEA3-A), all the other families (transformed with OsLEA3-S and OsLEA3-H
constructs) had higher grain yield than the wild type under drought stress in
both the field and the PVC pipes conditions. No significant yield penalty was
detected for these T(2 )and T(3) families. These results indicate that transgenic
rice with significantly enhanced drought resistance and without yield penalty can
be generated by over-expressing OsLEA3-1 gene with appropriate promoters and
following a bipartite (stress and non-stress) in-field screening protocol.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17426956 [PubMed - indexed for MEDLINE]

660: Genome. 2006 Dec;49(12):1528-38.

Loci affecting flowering time in oat under short-day conditions.

Locatelli AB, Federizzi LC, Milach SC, Wight CP, Molnar SJ, Chapados JT, Tinker
NA.

Federal University of Rio Grande do Sul, Departamento de Plantas de Lavoura,
Caixa postal 15100, CEP 91.501-970, Porto Alegre, RS, Brazil.

Flowering time (or days to heading) is an important characteristic in crop plants
that affects adaptation to cropping cycles and growing seasons. The objectives of
this study were to identify molecular markers associated with flowering time in 3
oat populations developed from Brazilian oat varieties, and to compare their map
locations with those of other loci that might influence flowering time. Flowering
time was studied in recombinant inbred lines from 3 hexaploid oat populations:
UFRGS 8 x Pc68/5*Starter; UFRGS 881971 x Pc68/5*Starter; and UFRGS 8 x UFRGS
930605. Bulked segregant analysis, using amplified fragment length polymorphism,
was followed by selective mapping in each population and in a reference
population, 'Kanota' x 'Ogle' (KxO). One quantitative trait locus (QTL) with
major effects on flowering time was identified in each cross. Comparative mapping
showed that a major QTL, with earliness alleles originating from UFRGS 8 and
UFRGS 881971, is in a region with close homology to KxO linkage group 17 and to a
locus that reportedly confers day-length insensitivity in oat (Di1). This is the
first report to identify the map location of the Di1 locus, and putatively
confirm the presence of Di1 alleles in new germplasm. Further comparative mapping
and the alignment of mapped oat markers with the sequenced rice genome suggest
that this QTL and (or) Di1 is orthologous to the Hd1 locus in rice and the
CONSTANS gene in Arabidopsis and other species. A different QTL with major
effects segregated in the UFRGS 8 x UFRGS 930605 cross, where the early-flowering
allele for Di1 was probably fixed. Two additional QTLs with smaller effects were
identified in the UFRGS 8 x Pc68/5*Starter population. These results suggest that
the Brazilian oat line UFRGS 8 contains an optimal set of alleles conditioning
earliness under the short-day conditions of the Brazilian winter growing season,
and that molecular selection could be used to introgress these alleles into other
breeding material.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17426767 [PubMed - indexed for MEDLINE]

661: Genome. 2006 Dec;49(12):1510-27.

Seed and agronomic QTL in low linolenic acid, lipoxygenase-free soybean (Glycine
max (L.) Merrill) germplasm.

Reinprecht Y, Poysa VW, Yu K, Rajcan I, Ablett GR, Pauls KP.

Department of Plant Agriculture, University of Guelph, Guelph, ON N1G 2W1,
Canada.

Linolenic acid and seed lipoxygenases are associated with off flavours in soybean
products. F5 recombinant inbred lines (RILs) from a cross between a low linolenic
acid line (RG10) and a seed lipoxygenase-free line (OX948) were genotyped for
simple sequence repeats (SSR), random amplified polymorphic DNA (RAPD),
sequence-tagged sites (STS), and cleaved amplified polymorphic sequence (CAPS)
markers and evaluated for seed and agronomic traits at 3 Ontario locations in 2
years. One hundred twenty markers covering 1247.5 cM were mapped to 18 linkage
groups (LGs) in the soybean composite genetic map. Seed lipoxygenases L-1 and L-2
mapped as single major genes to the same location on LG G13-F. L-3 mapped to LG
G11-E. This is the first report of a map position for L-3. A major quantitative
trait locus (QTL) associated with reduced linolenic acid content was identified
on LG G3-B2. QTLs for 12 additional seed and agronomic traits were detected.
Linolenic acid content, linoleic acid content, yield, seed mass, protein content,
and plant height QTL were present in at least 4 of 6 environments. Three to 8
QTLs per trait were detected that accounted for up to 78% of total variation.
Linolenic acid and lipoxygenase loci did not overlap yield QTL, suggesting that
it should be possible to develop high-yielding lines resistant to oxidative
degradation by marker-assisted selection (MAS).

PMID: 17426766 [PubMed - indexed for MEDLINE]

662: Autophagy. 2007 Jul-Aug;3(4):360-2. Epub 2007 Jul 19.

Comment on:
Plant Cell Physiol. 2006 Dec;47(12):1641-52.

Constitutive autophagy in plant root cells.

Yano K, Suzuki T, Moriyasu Y.

Graduate School of Nutritional and Environmental Sciences, University of
Shizuoka, Shizuoka, Japan.

In previous studies, using a membrane-permeable protease inhibitor, E-64d, we
showed that autophagy occurs constitutively in the root cells of barley and
Arabidopsis. In the present study, a fusion protein composed of the
autophagy-related protein AtAtg8 and green fluorescent protein (GFP) was
expressed in Arabidopsis to visualize autophagosomes. We first confirmed the
presence of autophagosomes with GFP fluorescence in the root cells of seedlings
grown on a nutrient-sufficient medium. The number of autophagosomes changed as
the root cells grew and differentiated. In cells near the apical meristem,
autophagosomes were scarcely found. However, a small but significant number of
autophagosomes existed in the elongation zone. More autophagosomes were found in
the differentiation zone where cell growth ceases but the cells start to form
root hair. In addition, we confirmed that autophagy is activated under starvation
conditions in Arabidopsis root cells. When the root tips were cultured in a
sucrose-free medium, the number of autophagosomes increased in the elongation and
differentiation zones, and a significant number of autophagosomes appeared in
cells near the apical meristem. The results suggest that autophagy in plant root
cells is involved not only in nutrient recycling under nutrient-limiting
conditions but also in cell growth and root hair formation.

Publication Types:
Comment

PMID: 17426438 [PubMed - indexed for MEDLINE]

663: Wei Sheng Yan Jiu. 2007 Jan;36(1):45-8.

[Expression and identification of Cry 1Ie in Bacillus coli]

[Article in Chinese]

Xu H, Zhang X, Li F, Wang G.

Institute of Nutrition and Food Safety, Chinese Center for Disease Control and
Prevention, Beijing.

OBJECTIVE: In this study, Cry1Ie expressed protein in the transgenic Cry1Ie maize
was used as a model protein to establish a technical platform for the safety
assessment of other genetically modified plants. METHODS: Bacillus coli
expression system was used to express Cry1Ie protein and then the expressed
protein was purified by SDS-PAGE. The equivalence of Cry1Ie proteins expressed in
Bacillus coli and in transgenic Cry1Ie maize was analyzed by their
immunorecognition and bioactivity. RESULTS: Cry1Ie protein was highly expressed
in Bacillus coli and could be segregated and purified by SDS-PAGE. Cry1Ie
expressed protein in Bacillus coli is substantially equivalent to that of maize
genetically modified with Cry1Ie in tests of immunorecognition western blot and
bioactivity in target pest. CONCLUSION: The established technical platform of
external Cry1Ie expressed protein can be applied to assess the food safety of
genetically modified plants.

Publication Types:
English Abstract

PMID: 17424848 [PubMed - in process]

664: Rev Med Univ Navarra. 2006 Oct-Dec;50(4):62-70.

[New challenges in basic and applied nutrition]

[Article in Spanish]

Palou A.

Laboratorio de Biologö­a Molecular, Nutriciö¨n y Biotecnologö­a (Nutrigenö¨mica),
Universitat de les Illes Balears, Palma de Mallorca.

The quality standards for nutrition and food have undergone major changes during
recent years in parallel with the increase in scientific knowledge in the area of
food and health. Trends in consumer demands are changing accordingly. While
during the last century our concerns were firstly focused on ensuring the
availability of basic foods and later on ensuring its safety, nowadays our
society is concentrating on improving wellbeing and, particularly, on tackling
and preventing the major chronic diseases such as cardiovascular diseases,
diabetes, obesity, diverse types of cancer, osteoporosis and autoimmune diseases.
These problems account for almost 50% of the diseases and 60% of deaths over the
world, and are directly related with the food we eat, irrespective of the
importance of other factors (age, sex, physical exercise, genetic predisposition,
nicotine poisoning or alcoholism). Europe has decided to go ahead with a major
legislative change in the food sector (the Regulation on nutrition and health
claims made on foods) which opens new perspectives to improve public health and
offer clear economic growth for the health-related food sector. Putting into
practice this new legislation and its consequences constitutes a major challenge.
Nevertheless, new challenges are foreseen: our response to food depends on our
individual genetic characteristics (nutrigenetics); it also depends on the
individual history that is being imprinted (in a permanent or temporary form) on
our chromosomes (epigenetics), as determined by the individual life style (food,
different episodes and facts, including emotions) and, particularly, nutrition
during the most active stages of our development. For the future, the additional
contribution we will ask of food is to promote health and well-being in all
facets. The requirement is that it should develop in a free framework, based on
the best scientific available advice, with transparency as a fundamental
guarantee. In this context, new disciplines (e.g. nutrigenomics) are emerging and
new trends towards a more conscious, better informed, consumer are foreseen that
will allow an appropriate reaction to these new challenges in the medium and long
term.

Publication Types:
English Abstract
Review

PMID: 17424771 [PubMed - indexed for MEDLINE]

665: Plant Mol Biol. 2007 Jul;64(4):453-66. Epub 2007 Apr 10.

The SebHLH transcription factor mediates trans-activation of the SeFAD2 gene
promoter through binding to E- and G-box elements.

Kim MJ, Kim JK, Shin JS, Suh MC.

School of Life Sciences and Biotechnology, Korea University, Seoul, 136-701,
Korea.

Microsomal oleic acid desaturase (FAD2) catalyzes the first extra-plastidial
desaturation in plants, converting oleic acid to linoleic acid, which is a major
constituent in all cellular membranes as well as in seed storage oils.
Seed-specific FAD2 (SeFAD2) produced 40% of linoleic acids in the total fatty
acids of sesame (Sesamum indicum) seeds. The expression of SeFAD2 transcripts was
spatially and temporally controlled during seed development. To investigate the
regulatory mechanism controlling seed-specific SeFAD2 expression, we isolated a
well-matched sequence homologous to the basic region/helix-loop-helix proteins by
yeast one-hybrid screening and named it SebHLH. SebHLH transcripts were expressed
in developing seeds and roots of sesame. SebHLH:GFP fusion protein localized in
the nucleus. Recombinant SebHLH protein bound E-box (CANNTG) and G-box (CACGTG)
elements in the region from -179 to -53 of the SeFAD2 gene promoter, and the
external C and G nucleotides in the E- and G-box motifs were essential for SebHLH
protein binding. The SebHLH gene, under the CaMV35S promoter, and the GUS
reporter gene driven by E- and G-box motifs were co-expressed in developing
sesame seeds and Arabidopsis transgenic leaves. This co-expression demonstrated
that SebHLH protein mediates transactivation of the SeFAD2 gene promoter through
binding to E- and G-box elements. E- or G-box elements frequently occur in the
5'-flanking region of genes that are involved in triacylglycerol biosynthesis and
that exhibit seed-specific expression in Arabidopsis and other plants, suggesting
that bHLH transcription factors play a key role in the transcriptional regulation
of genes related to storage lipid biosynthesis and accumulation during seed
development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17420955 [PubMed - indexed for MEDLINE]

666: Nat Biotechnol. 2007 Apr;25(4):367-8. Epub 2007 Apr 3.

US courts thwart GM alfalfa and turf grass.

Fox JL.

Publication Types:
News

PMID: 17420729 [PubMed - indexed for MEDLINE]

667: Biosci Biotechnol Biochem. 2007 Apr;71(4):1107-10. Epub 2007 Apr 7.

Characterization of the rice circadian clock-associated pseudo-response
regulators in Arabidopsis thaliana.

Murakami M, Tago Y, Yamashino T, Mizuno T.

Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University,
Nagoya, Japan. t2706775@mbox.nagoya-u.ac.jp

Members of the small family of Arabidopsis PSEUDO-RESPONSE REGULATORS (PRR1/TOC1,
PRR3, PRR5, PRR7, and PRR9) play roles close to the circadian clock in
Arabidopsis thaliana. We have reported that the rice (Oryza sativa) genome also
encodes a set of PRR counterparts (designated OsPRR1, OsPRR37, OsPRR59, OsPRR73,
and OsPRR95 respectively). To gain new insight into the molecular functions of
OsPRRs, we carried out genetic complementation analyses by introducing two
representative rice genes, OsPRR1 and OsPRR37, into the corresponding Arabidopsis
loss-of-function mutants (toc1 and prr7 respectively). The results showed that
these OsPRR and AtPRR genes are genetically interchangeable at least in part,
suggesting the conserved clock-associated function of these OsPRRs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17420570 [PubMed - indexed for MEDLINE]

668: Plant J. 2007 Jun;50(5):825-38. Epub 2007 Apr 5.

WRINKLED1 specifies the regulatory action of LEAFY COTYLEDON2 towards fatty acid
metabolism during seed maturation in Arabidopsis.

Baud S, Mendoza MS, To A, Harscoö«t E, Lepiniec L, Dubreucq B.

Laboratoire de Biologie des Semences, IJPB,UMR 204 INRA/AgroParis Tech, F-78026
Versailles, France.

The WRINKLED1 (WRI1) transcription factor has been shown to play a role of the
utmost importance during oil accumulation in maturing seeds of Arabidopsis
thaliana. However, little is known about the regulatory processes involved. In
this paper, comprehensive functional analyses of three new mutants corresponding
to null alleles of wri1 confirm that the induction of WRI1 is a prerequisite for
fatty acid synthesis and is important for normal embryo development. The strong
expression of WRI1 specifically detected at the onset of the maturation phase in
oil-accumulating tissues of A. thaliana seeds is fully consistent with this
function. Complementation experiments carried out with various seed-specific
promoters emphasized the importance of a tight regulation of WRI1 expression for
proper oil accumulation, raising the question of the factors controlling WRI1
transcription. Interestingly, molecular and genetic analyses using an inducible
system demonstrated that WRI1 is a target of LEAFY COTYLEDON2 and is necessary
for the regulatory action of LEC2 towards fatty acid metabolism. In addition to
this, quantitative RT-PCR experiments suggested that several genes encoding
enzymes of late glycolysis, the fatty acid synthesis pathway, and the biotin and
lipoic acid biosynthetic pathways are targets of WRI1. Taken together, these
results indicate new relationships in the regulatory model for the control of oil
synthesis in maturing A. thaliana seeds. In addition, they exemplify how
metabolic and developmental processes affecting the developing embryo can be
coordinated at the molecular level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17419836 [PubMed - indexed for MEDLINE]

669: Anal Chim Acta. 2007 Apr 25;589(2):159-65. Epub 2007 Feb 25.

Solubilisation and binding characteristics of a recombinant beta2-adrenergic
receptor expressed in the membrane of Escherichia coli for the multianalyte
detection of beta-agonists and antagonists residues in food-producing animals.

Danyi S, Degand G, Duez C, Granier B, Maghuin-Rogister G, Scippo ML.

Department of Food Sciences, Laboratory of Analysis, Faculty of Veterinary
Medicine, University of Liö¨ge, Sart-Tilman, Liö¨ge, Belgium. sdanyi@ulg.ac.be


The number of substances with beta-agonistic activity, illegally introduced in
meat production or in sports doping as anabolic or beta-blocking agents is
increasing. Analytical methods suited for their multianalyte detection are thus
necessary. In this perspective, receptor assays were developed. The research
activities undertaken in this study describe the solubilisation of a recombinant
human beta(2)-adrenergic receptor produced in the inner membrane of genetically
modified Escherichia coli, using the detergent n-dodecyl-beta-d-maltoside. Its
potential to detect the presence of beta-agonists or beta-blockers in biological
samples was evaluated. The solubilised beta(2)-adrenergic receptor retained its
binding affinity in a radio-receptor assay based on the competition for the
binding to receptors between a ligand (beta-agonist or antagonist) and the
radioligand [(125)I]iodocyanopindolol. The IC(50) values ranged from 5+/-1 x
10(-8) M (clenbuterol) to 8+/-2 x 10(-6) M (isoxsuprine) for the beta-agonists
tested and from 1.5+/-0.2 x 10(-10) M (carazolol) to 1.2+/-0.2 x 10(-5) M
(metoprolol) for the beta-blockers tested. It was shown to have a lower limit of
detection than a radio-receptor assay using the solubilised beta(2)-adrenoceptor
expressed in a mammalian cell line. The solubilised recombinant human
beta(2)-adrenoreceptor expressed in E. coli would be a useful tool to develop non
radioactive multianalyte screening methods.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17418176 [PubMed - indexed for MEDLINE]

670: J Agric Food Chem. 2007 May 2;55(9):3421-8. Epub 2007 Apr 7.

Oral toxicity of beta-N-acetyl hexosaminidase to insects.

Dowd PF, Johnson ET, Pinkerton TS.

Crop BioProtection Research Unit, Agricultural Research Service, U.S. Department
of Agriculture, National Center for Agricultural Utilization Research, 1815 North
University Street, Peoria, Illinois 61604, USA. partick.dowd@ars.usda.gov

Insect chitin is a potential target for resistance plant proteins, but
plant-derived chitin-degrading enzymes active against insects are virtually
unknown. Commercial beta-N-acetylhexosaminidase (NAHA), a chitin-degrading enzyme
from jack bean Canavalia ensiformis, caused significant mortality of fall
armyworm Spodoptera frugiperda larvae at 75 microg/gm, but no significant
mortality was noted with Aspergillus niger NAHA. Maize Zea mays callus
transformed to express an Arabidopsis thaliana clone that putatively codes for
NAHA caused significantly higher mortality of cigarette beetle Lasioderma
serricorne larvae and significantly reduced growth rates (as reflected by
survivor weights) of S. frugiperda as compared to callus that expressed control
cDNAs. Tassels from model line Hi-II maize (Z. mays) plants transformed with the
NAHA gene fed to S. frugiperda caused significantly higher mortality than tassels
transformed to express glucuronidase; mortality was significantly correlated with
NAHA expression levels detected histochemically. Leaf disks from inbred Oh43
maize plants transformed with the NAHA gene on average had significantly less
feeding by caterpillars than null transformants. Leaf disks of Oh43 transformants
caused significant mortality of both S. frugiperda and corn earworm Helicoverpa
zea larvae, which was associated with higher expression levels of NAHA detected
by isoelectric focusing, histochemically, or with antibody. Overall, these
results suggest that plant NAHA has a role in insect resistance. Introduction of
NAHA genes or enhancement of activity through breeding or genetic engineering has
the potential to significantly reduce insect damage and thereby indirectly reduce
mycotoxins that are harmful to animals and people.

PMID: 17417870 [PubMed - indexed for MEDLINE]

671: Transgenic Res. 2007 Dec;16(6):795-812. Epub 2007 Apr 6.

Bitrophic and tritrophic effects of Bt Cry3A transgenic potato on beneficial,
non-target, beetles.

Ferry N, Mulligan EA, Majerus ME, Gatehouse AM.

School of Biology, Institute for Research on Environment and Sustainability,
University of Newcastle Upon Tyne, Devonshire Building, Newcastle NE1 7RU, UK.

Insect-resistant transgenic plants have been suggested to have unpredictable
effects on the biodiversity of the agro-ecosystem, including potential effects on
insect natural enemies, beneficial in control of crop pests. Whilst carnivorous
as adults, many of these predators may also consume plant tissues, in particular
plant pollen and nectar. Coleoptera are important in terms of agro-ecological
research not only because of the large number of species in this order, but also
because of their role as biological control agents. Thus any detrimental impact
on this group of insects would be highly undesirable. The effects of potato
expressing the coleopteran-specific Bacillus thuringiensis delta-endotoxin Cry3A
(Bt Cry3A) on the ladybird beetle Harmonia axyridis and the carabid beetle Nebria
brevicollis were investigated via the bitrophic interaction of the adult ladybird
with potato flowers and the tritrophic interaction of the carabid consuming a
non-target potato pest. Immunoassays confirmed accumulation of the transgene
product in potato leaves and floral tissues (at levels of up to 0.01% (pollen)
and 0.0285% (anthers) of total soluble protein). Despite H. axyridis and N.
brevicollis belonging to the targeted insect order, no significant effects upon
survival or overall body mass change of either beetle were observed. Furthermore,
Bt Cry3A had no detrimental effects on reproductive fitness of either beetle
species, either in terms of fecundity or subsequent egg viability. Behavioural
analysis revealed no significant impact of Bt Cry3A on beetle activity or
locomoter behaviour. Ligand blots indicate that this is due to either the absence
of Bt-binding sites in brush border membrane vesicles (BBMV) isolated from Nebria
brevicollis, or in the case of Harmonia axyridis, the binding did not
functionally lead to behavioural or physical effects.

Publication Types:
Comparative Study

PMID: 17415673 [PubMed - indexed for MEDLINE]

672: Biotechnol Lett. 2007 May;29(5):829-35. Epub 2007 Feb 16.

A phosphate starvation-induced acid phosphatase from Oryza sativa: phosphate
regulation and transgenic expression.

Hur YJ, Lee HG, Jeon EJ, Lee YY, Nam MH, Yi G, Eun MY, Nam J, Lee JH, Kim DH.

College of Natural Resources and Life Science, Dong-A University, Busan, 604-714,
Korea.

A phosphate starvation-induced acid phosphatase cDNA was cloned from the rice,
Oryza sativa. The cDNA encoding O. sativa acid phosphatase (OsACP1) has 1100 bp
with an open reading frame of 274 amino acid residues. The deduced amino acid
sequence of OsACP1 cDNA showed 53% identity to tomato acid phosphatase and 46-50%
identity to several other plant phosphatases. OsACP1 expression was up-regulated
in the rice plant and in cell culture in the absence of phosphate (Pi). The
induced expression of OsACP1 was a specific response to Pi starvation, and was
not affected by the deprivation of other nutrients. OsACP1 expression was
responsive to the level of Pi supply, with transcripts of OsACP1 being abundant
in Pi-deprived root. The OsACP1 cDNA was expressed as a 30 kDa polypeptide in
baculovirus-infected insect Sf9 cells. In addition, the OsACP1 gene was
introduced into Arabidopsis via Agrobacterium-mediated transformation. Functional
expression of the OsACP1 gene in the transgenic Arabidopsis lines was confirmed
by Northern blot and Western blot analyses, as well as phosphatase activity
assays. These results suggest that the OsACP1 gene can be used to develop new
transgenic dicotyledonous plants able to adapt to Pi-deficient conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17415667 [PubMed - indexed for MEDLINE]

673: Bull Entomol Res. 2007 Apr;97(2):211-5.

Evaluation of Bt-toxin uptake by the non-target herbivore, Myzus persicae
(Hemiptera: Aphididae), feeding on transgenic oilseed rape.

Burgio G, Lanzoni A, Accinelli G, Dinelli G, Bonetti A, Marotti I, Ramilli F.

Dipartimento di Scienze e Tecnologie Agroambientali, Alma Mater
Studiorum-Universitö  di Bologna, Viale Fanin 42, 40127 Bologna, Italy.
gburgio@entom.agrsci.unibo.it

As consequence of the concern about the biosafety of genetically modified plants,
biological and ecological studies are considered crucial for environmental risk
assessment. Laboratory experiments were carried out in order to evaluate the
transfer of the Cry1Ac Bt-toxin from a transgenic Bt-oilseed rape to a non-target
pest, Myzus persicae Sulzer. Cry1Ac protein levels in plants and aphids were
determined using a double sandwich enzyme-linked immunosorbent assay. Phloem sap
from (Bt+) and (Bt-) oilseed rape plants was collected from leaves using a
standard method of extraction in an EDTA buffer. Bt-toxin was present in phloem
sap, with a mean concentration of 2.7 +/- 1.46 ppb, corresponding to a 24-fold
lower level than in oilseed rape leaves. Toxin was also detected in aphid
samples, with a mean concentration in the positive samples of 2.0 +/- 0.8 ppb.
The evidence that Bt-toxin remains in herbivores, in this case an aphid, could be
useful to clarify functional aspects linked to possible consequences of Bt-crops
on food chains involving herbivore-natural enemy trophic systems. Further studies
are needed in order to improve the knowledge on the functional aspects linked to
the transfer of the Cry1Ac Bt-toxin from GM-oilseed rape to aphids and their
possible consequence.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17411484 [PubMed - indexed for MEDLINE]

674: J Agric Food Chem. 2007 May 2;55(9):3304-11. Epub 2007 Apr 6.

Identification and quantification of stilbenes in fruits of transgenic tomato
plants (Lycopersicon esculentum Mill.) by reversed phase HPLC with photodiode
array and mass spectrometry detection.

Nicoletti I, De Rossi A, Giovinazzo G, Corradini D.

Istituto di Metodologie Chimiche - CNR, Area della Ricerca di Roma 1, Via Salaria
Km 29,300 Montelibretti, P.O. Box 10, 00016 Monterotondo Stazione (Rome), Italy.

Reversed-phase high-performance liquid chromatography (RP-HPLC) with photodiode
array (PDA) and mass spectrometry (MS) detection was employed to study the
accumulation of stilbenes and other naturally occurring polyphenol intermediates
of flavonoid pathway in tomato fruits of plants genetically modified to
synthesize resveratrol. The transgenic tomato fruits were obtained by
overexpression of a grapevine gene encoding the enzyme stilbene synthase in
tomato plants (Lycopersicon esculentum Mill.). Stilbenes and flavonoids, either
glycosylated or free, were simultaneosly identified by electrospray interface
(ESI)-MS in negative ionization mode and were quantified by PDA detection at the
wavelength corresponding to their maximum absorbance. The two detectors were
coupled online with an HPLC system utilizing a narrow-bore C18 reversed-phase
column, which was eluted by a multistep gradient of increasing concentration of
acetonitrile in water containing 0.5% (v/v) formic acid. The results of these
analysis revealed that the genetic modification of the tomato plants originated
different levels of accumulation of four stilbenes (i.e., trans- and cis-piceid
and trans- and cis-resveratrol) in their fruit depending on the stages of
ripening. Either at immature or at mature stages of ripening the stilbenes were
preferentially accumulated in the fruit peel as the glycosylated form. The
highest amount of trans-piceid and trans-resveratrol were found in the peel of
fruits harvested at mature stage of ripening. The variations in the levels of
rutin, naringenin, and chlorogenic acid found in the samples extracted from the
fruits of transgenic tomato plants, in comparison to that determined in the
control lines, seemed to be related to the genetic transformation, whose effect
on the flavonoid biosynthetic pathway needs to be elucidated by additional
studies.

PMID: 17411064 [PubMed - indexed for MEDLINE]

675: Environ Sci Technol. 2007 Mar 15;41(6):1863-9.

Selenium volatiles as proxy to the metabolic pathways of selenium in genetically
modified Brassica juncea.

Kubachka KM, Meija J, LeDuc DL, Terry N, Caruso JA.

Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172,
USA.

In this study we demonstrate that the headspace selenium volatiles could be used
as proxy to the metabolic pathways in the Se-accumulator plant Brassica juncea.
The selenium metabolic pathways in wild type plants are compared to those of
several genetically modified cultures. Complementary use of atomic and molecular
mass spectrometric techniques also allowed for identification of yet unreported
minor headspace Se-containing volatiles such as CH3SeSeSeCH3, CH3SeSSeCH3, and
CH3SeCH2CH3. By combining the information resulting from this research with the
previously known information about selenium metabolism in B. juncea, it is
possible that a more efficacious phytoremediation tool can be constructed.

Publication Types:
Comparative Study
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17410776 [PubMed - indexed for MEDLINE]

676: Annu Rev Phytopathol. 2007;45:173-202.

Safety of virus-resistant transgenic plants two decades after their introduction:
lessons from realistic field risk assessment studies.

Fuchs M, Gonsalves D.

Department of Plant Pathology, Cornell University, New York State Agricultural
Experiment Station, Geneva, NY 14456, USA. mf13@cornell.edu

Potential safety issues have been raised with the development and release of
virus-resistant transgenic plants. This review focuses on safety assessment with
a special emphasis on crops that have been commercialized or extensively tested
in the field such as squash, papaya, plum, grape, and sugar beet. We discuss
topics commonly perceived to be of concern to the environment and to human
health--heteroencapsidation, recombination, synergism, gene flow, impact on
nontarget organisms, and food safety in terms of allergenicity. The wealth of
field observations and experimental data is critically evaluated to draw
inferences on the most relevant issues. We also express inside views on the
safety and benefits of virus-resistant transgenic plants, and recommend realistic
risk assessment approaches to assist their timely deregulation and release.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Review

PMID: 17408355 [PubMed - indexed for MEDLINE]

677: J Agric Food Chem. 2007 May 2;55(9):3268-74. Epub 2007 Apr 4.

Toward metrological traceability for DNA fragment ratios in GM quantification. 3.
Suitability of DNA calibrants studied with a MON 810 corn model.

Charels D, Broeders S, Corbisier P, Trapmann S, Schimmel H, Emons H.

European Commission, Joint Research Centre, Institute for Reference Materials and
Measurements, IRMM, Retieseweg 111, 2440 Geel, Belgium.
diana.charels@ec.europa.eu

The quantification of GMOs by real-time PCR relies on an external calibrant. In
this paper the suitability of two DNA calibrants, genomic DNA from plant leaves
and plasmidic DNA, was investigated. The PCR efficiencies, the correlation
coefficients of the calibration curves, and the ratios between PCR efficiencies
of transgenic and endogenous sequences were compared for both calibrants using 59
data sets produced by 43 laboratories. There were no significant differences
between plasmidic and genomic DNA except for the PCR efficiencies of the
calibration curves for the transgene of the construct-specific real-time PCR
method. In the GM system investigated, PCR efficiencies of plasmidic calibrants
were slightly closer to the PCR efficiencies observed for the unknowns than those
of the genomic DNA calibrant. Therefore, plasmidic DNA was the more suitable
calibrant for the PCR measurements on genomic DNA extracted from MON 810 seeds.
It is shown that plasmidic DNA is an appropriate choice for the calibration of
measurements of MON 810 corn with respect to the DNA copy number ratio.

PMID: 17407307 [PubMed - indexed for MEDLINE]

678: J Agric Food Chem. 2007 May 2;55(9):3258-67. Epub 2007 Apr 4.

Toward metrological traceability for DNA fragment ratios in GM quantification. 2.
Systematic study of parameters influencing the quantitative determination of MON
810 corn by real-time PCR.

Charels D, Broeders S, Corbisier P, Trapmann S, Schimmel H, Linsinger T, Emons H.

European Commission, Joint Research Centre, Institute for Reference Materials and
Measurements (IRMM), Retieseweg 111, 2440 Geel, Belgium.
diana.charels@ec.europa.eu

This paper is part of a set of three papers investigating metrological
traceability of the quantification of DNA fragments as, for instance, used for
quantification of genetic modifications. This paper evaluates the possible impact
of several factors on results of real-time Polymerase Chain Reaction (PCR)
measurements. It was found that the particle size of the powder samples does not
have an influence, whereas the nature of the calibrant (plasmidic or genomic DNA)
has a significant effect. Moreover, two real-time PCR detection methods
(construct-specific and event-specific) for MON 810 corn were compared. The
results obtained in a specifically designed interlaboratory study revealed a
significant influence of the DNA extraction method on measurement results when
the MON 810 construct-specific real-time PCR detection method was applied.
Statistical analyses confirmed the importance of validating DNA extraction
methods in conjunction with real-time PCR methods.

PMID: 17407306 [PubMed - indexed for MEDLINE]

679: J Agric Food Chem. 2007 May 2;55(9):3249-57. Epub 2007 Apr 4.

Toward metrological traceability for DNA fragment ratios in GM quantification. 1.
Effect of DNA extraction methods on the quantitative determination of Bt176 corn
by real-time PCR.

Corbisier P, Broothaerts W, Gioria S, Schimmel H, Burns M, Baoutina A, Emslie KR,
Furui S, Kurosawa Y, Holden MJ, Kim HH, Lee YM, Kawaharasaki M, Sin D, Wang J.

European Commission, Joint Research Centre, Institute for Reference Materials and
Measurements (IRMM), Retieseweg 111, 2440 Geel, Belgium.
philippe.corbisier@ec.europa.edu.

An international CCQM-P60 pilot study involving eight national metrological
institutes was organized to investigate if the quantification of genetically
modified (GM) corn powder by real-time PCR was affected by the DNA extraction
method applied. Four commonly used extraction methods were compared for the
extraction of DNA from a GM Bt176 corn powder. The CTAB-based method yielded the
highest DNA template quantity and quality. A difference in the 260 nm/230 nm
absorbance ratio was observed among the different extraction methods. Real-time
amplification of sequences specific for endogenous genes zein and hmg as well as
transgenic sequences within the cryIA(b) gene and a fragment covering the
junction between the transformed DNA and the plant genome were used to determine
the GM percentage. The detection of the transgenic gene was affected by the
quantity and quality of template used for the PCR reaction. The Bt176 percentages
measured on diluted or purified templates were statistically different depending
on the extraction method applied.

PMID: 17407305 [PubMed - indexed for MEDLINE]

680: Plant Cell Rep. 2007 Aug;26(8):1253-62. Epub 2007 Apr 4.

A strong constitutive gene expression system derived from ibAGP1 promoter and its
transit peptide.

Kwak MS, Oh MJ, Lee SW, Shin JS, Paek KH, Bae JM.

School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, South
Korea.

To develop a strong constitutive gene expression system, the activities of ibAGP1
promoter and its transit peptide were investigated using transgenic Arabidopsis
and a GUS reporter gene. The ibAGP1 promoter directed GUS expression in almost
entire tissues including rosette leaf, inflorescence stem, inflorescence, cauline
leaf and root, suggesting that the ibAGP1 promoter is a constitutive promoter.
GUS expression mediated by ibAGP1 promoter was weaker than that by CaMV35S
promoter in all tissue types, but when GUS protein was targeted to plastids with
the aid of the ibAGP1 transit peptide, GUS levels increased to higher levels in
lamina, petiole and cauline leaf compared to those produced by CaMV35S promoter.
The enhancing effect of ibAGP1 transit peptide on the accumulation of foreign
protein was tissue-specific; accumulation was high in lamina and inflorescence,
but low in root and primary inflorescence stem. The transit peptide effect in the
leaves was maintained highly regardless of developmental stages of plants. The
ibAGP1 promoter and its transit peptide also directed strong GUS gene expression
in transiently expressed tobacco leaves. These results suggest that the ibAGP1
promoter and its transit peptide are a strong constitutive foreign gene
expression system for transgenesis of dicot plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17406871 [PubMed - indexed for MEDLINE]

681: Mol Genet Genomics. 2007 Jul;278(1):85-94. Epub 2007 Apr 3.

The rice OsLOL2 gene encodes a zinc finger protein involved in rice growth and
disease resistance.

Xu C, He C.

State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese
Academy of Sciences, Beijing, 100080, People's Republic of China.

Arabidopsis LSD1-related proteins that contain LSD1-like zinc finger domains have
been identified to be involved in disease resistance and programmed cell death.
To investigate the potential role of LSD1-related gene in rice (Oryza sativa L.),
we cloned an LSD1 ortholog, OsLOL2, from the rice cDNA plasmid library. The
OsLOL2 gene is predicted to encode a polypeptide of 163 amino acids with two
LSD1-like zinc finger domains with 74.5% identity to those of LSD1. Southern blot
analysis indicated that OsLOL2 was a single-copy gene in the rice genome.
Transgenic rice lines carrying the antisense strand of OsLOL2 with decreased
expression of OsLOL2 had dwarf phenotypes, and the dwarfism could be restored by
exogenous GA(3) treatment, suggesting that the dwarfism was the result of a
deficiency in bioactive gibberellin (GA). In agreement with this possibility, the
content of endogenous bioactive GA(1) decreased in the antisense transgenic
lines. Expression of OsKS1, one of the genes encoding for GA biosynthetic
enzymes, was suppressed in the antisense transgenic lines. Sense transgenic lines
with increased expression of OsLOL2 were more resistant to rice bacterial blight,
while antisense transgenic lines were less resistant to rice bacterial blight.
The OsLOL2-GFP (green fluorescence protein) fusion protein was localized in the
nucleus of cells of transgenic BY2 tobacco (Nicotiana tabacum L.). These data
suggest that OsLOL2 is involved in rice growth and disease resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17404758 [PubMed - indexed for MEDLINE]

682: J Agric Food Chem. 2007 May 2;55(9):3351-7. Epub 2007 Apr 3.

Detection methods for biotech cotton MON 15985 and MON 88913 by PCR.

Lee SH, Kim JK, Yi BY.

Gene Analysis Laboratory, Experiment Research Institute of National Agricultural
Products Quality Management Service, Seoul 150-043, South Korea.
starlee@naqs.go.kr

Plants derived through agricultural biotechnology, or genetically modified
organisms (GMOs), may affect human health and ecological environment. A living
GMO is also called a living modified organism (LMO). Biotech cotton is a GMO in
food or feed and also an LMO in the environment. Recently, two varieties of
biotech cotton, MON 15985 and MON 88913, were developed by Monsanto Co. The
detection method is an essential element for the GMO labeling system or LMO
management of biotech plants. In this paper, two primer pairs and probes were
designed for specific amplification of 116 and 120 bp PCR products from MON 15985
and MON 88913, respectively, with no amplification from any other biotech cotton.
Limits of detection of the qualitative method were all 0.05% for MON 15985 and
MON 88913. The quantitative method was developed using a TaqMan real-time PCR. A
synthetic plasmid, as a reference molecule, was constructed from a taxon-specific
DNA sequence of cotton and two construct-specific DNA sequences of MON 15985 and
MON 88913. The quantitative method was validated using six samples that contained
levels of biotech cotton mixed with conventional cotton ranging from 0.1 to
10.0%. As a result, the biases from the true value and the relative deviations
were all within the range of +/-20%. Limits of quantitation of the quantitative
method were all 0.1%. Consequently, it is reported that the proposed detection
methods were applicable for qualitative and quantitative analyses for biotech
cotton MON 15985 and MON 88913.

PMID: 17402745 [PubMed - indexed for MEDLINE]

683: EMBO Rep. 2007 Apr;8(4):305-8.

Comment in:
EMBO Rep. 2007 Apr;8(4):309-15. EMBO Rep. 2007 Jul;8(7):612-3.

The precautionary principle should not be used as a basis for decision-making.
Talking point on the precautionary principle.

Peterson M.

Department of History and Philosophy of Science, University of Cambridge, UK.
mbp24@cam.ac.uk

PMID: 17401402 [PubMed - indexed for MEDLINE]

684: Plant Cell Physiol. 2007 May;48(5):736-44. Epub 2007 Mar 31.

Antisense expression of 3-oxoacyl-ACP reductase affects whole plant productivity
and causes collateral changes in activity of fatty acid synthase components.

O'Hara P, Slabas AR, Fawcett T.

School of Biological and Biomedical Sciences, Durham University, South Road,
Durham DH1 3LE, UK.

Brassica napus cv Westar plants were transformed with 3-oxoacyl-ACP reductase
(KR) in antisense orientation, driven by either the cauliflower mosaic virus 35S
promoter or a seed-specific acyl carrier protein promoter to determine the
effects on plant productivity and on the activity of other fatty acid synthase
(FAS) components. In plants with altered KR activity, total seed yield was
reduced in all cases. In less severely affected plant lines, seeds had a normal
appearance and composition but the yield of seeds was reduced by approximately
50%. In more severely affected lines, reductions in both seed fatty acid content
and the number of seeds produced per plant were evident, resulting in a 90%
reduction in fatty acid synthesized per plant. These phenotypes were independent
of the promoter used. In severely affected lines, a large proportion of seeds
showed precocious germination, and these had a reduced oleate content and
increased levels of polyunsaturated 18-carbon fatty acids, compared with normal
seeds of the same line. This reduction in 18:1 fatty acids was mimicked on
imbibition of seeds with a normal appearance, indicating a preferential use of
oleate moieties in precocious germination events. The reduction in activity of KR
was mirrored for a second fatty acid synthase component, enoyl-ACP reductase,
indicating a mechanism to maintain the ratio of fatty acid synthase components
throughout embryogenesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17401135 [PubMed - indexed for MEDLINE]

685: Plant Cell. 2007 Mar;19(3):847-61. Epub 2007 Mar 30.

Rice UDP-glucose pyrophosphorylase1 is essential for pollen callose deposition
and its cosuppression results in a new type of thermosensitive genic male
sterility.

Chen R, Zhao X, Shao Z, Wei Z, Wang Y, Zhu L, Zhao J, Sun M, He R, He G.

Key Laboratory of Ministry of Education for Plant Developmental Biology, College
of Life Sciences, Wuhan University, Wuhan 430072, China.

UDP-glucose pyrophosphorylase (UGPase) catalyzes the reversible production of
glucose-1-phosphate and UTP to UDP-glucose and pyrophosphate. The rice (Oryza
sativa) genome contains two homologous UGPase genes, Ugp1 and Ugp2. We report a
functional characterization of rice Ugp1, which is expressed throughout the
plant, with highest expression in florets, especially in pollen during anther
development. Ugp1 silencing by RNA interference or cosuppression results in male
sterility. Expressing a double-stranded RNA interference construct in Ugp1-RI
plants resulted in complete suppression of both Ugp1 and Ugp2, together with
various pleiotropic developmental abnormalities, suggesting that UGPase plays
critical roles in plant growth and development. More importantly,
Ugp1-cosuppressing plants contained unprocessed intron-containing primary
transcripts derived from transcription of the overexpression construct. These
aberrant transcripts undergo temperature-sensitive splicing in florets, leading
to a novel thermosensitive genic male sterility. Pollen mother cells (PMCs) of
Ugp1-silenced plants appeared normal before meiosis, but during meiosis, normal
callose deposition was disrupted. Consequently, the PMCs began to degenerate at
the early meiosis stage, eventually resulting in complete pollen collapse. In
addition, the degeneration of the tapetum and middle layer was inhibited. These
results demonstrate that rice Ugp1 is required for callose deposition during PMC
meiosis and bridges the apoplastic unloading pathway and pollen development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17400897 [PubMed - indexed for MEDLINE]

686: Plant J. 2007 Apr;50(1):70-9.

Increase in CPD photolyase activity functions effectively to prevent growth
inhibition caused by UVB radiation.

Hidema J, Taguchi T, Ono T, Teranishi M, Yamamoto K, Kumagai T.

Department of Environmental Life Sciences, Graduate School of Life Sciences,
Tohoku University, Sendai 980-8577, Japan.

Rice cultivars vary widely in their sensitivity to ultraviolet B (UVB) and this
has been correlated with cyclobutane pyrimidine dimer (CPD) photolyase mutations
that alter the structure/function of this photorepair enzyme. Here, we tested
whether CPD photolyase function determines the UVB sensitivity of rice (Oryza
sativa) by generating transgenic rice plants bearing the CPD photolyase gene of
the UV-resistant rice cultivar Sasanishiki in the sense orientation (S-B and S-C
lines) or the antisense orientation (AS-D line). The S-B and S-C plants had 5.1-
and 45.7-fold higher CPD photolyase activities than the wild-type, respectively,
were significantly more resistant to UVB-induced growth damage, and maintained
significantly lower CPD levels in their leaves during growth under elevated UVB
radiation. Conversely, the AS-D plant had little photolyase activity, was
severely damaged by elevated UVB radiation, and maintained higher CPD levels in
its leaves during growth under UVB radiation. Notably, the S-C plant was not more
resistant to UVB-induced growth inhibition than the S-B plant, even though it had
much higher CPD photolyase activity. These results strongly indicate that
UVB-induced CPDs are one of principal causes of UVB-induced growth inhibition in
rice plants grown under supplementary UVB radiation, and that increasing CPD
photolyase activity can significantly alleviate UVB-caused growth inhibition in
rice. However, further protection from UVB-induced damage may require the genetic
enhancement of other systems as well.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17397507 [PubMed - indexed for MEDLINE]

687: J Fish Dis. 2007 Apr;30(4):201-12.

Evaluation of stress- and immune-response biomarkers in Atlantic salmon, Salmo
salar L., fed different levels of genetically modified maize (Bt maize), compared
with its near-isogenic parental line and a commercial suprex maize.

Sagstad A, Sanden M, Haugland ö˜, Hansen AC, Olsvik PA, Hemre GI.

National Institute of Nutrition and Seafood Research, NIFES, Bergen, Norway.

The present study was designed to evaluate if genetically modified (GM) maize (Bt
maize, event MON810) compared with the near-isogenic non-modified (nGM) maize
variety, added as a starch source at low or high inclusions, affected fish health
of post-smolt Atlantic salmon, Salmo salar L. To evaluate the health impact,
selected stress- and immune-response biomarkers were quantified at the gene
transcript (mRNA) level, and some also at the protein level. The diets with low
or high inclusions of GM maize, and its near-isogenic nGM parental line, were
compared to a control diet containing GM-free suprex maize (reference diet) as
the only starch source. Total superoxide dismutase (SOD) activity in liver and
distal intestine was significantly higher in fish fed GM maize compared with fish
fed nGM maize and with the reference diet group. Fish fed GM maize showed
significantly lower catalase (CAT) activity in liver compared with fish fed nGM
maize and to the reference diet group. In contrast, CAT activity in distal
intestine was significantly higher for fish fed GM maize compared with fish fed
reference diet. Protein level of heat shock protein 70 (HSP70) in liver was
significantly higher in fish fed GM maize compared with fish fed the reference
diet. No diet-related differences were found in normalized gene expression of
SOD, CAT or HSP70 in liver or distal intestine. Normalized gene expression of
interleukin-1 beta in spleen and head-kidney did not vary significantly between
diet groups. Interestingly, fish fed high GM maize showed a significantly larger
proportion of plasma granulocytes, a significantly larger sum of plasma
granulocyte and monocyte proportions, but a significantly smaller proportion of
plasma lymphocytes, compared with fish fed high nGM maize. In conclusion,
Atlantic salmon fed GM maize showed some small changes in stress protein levels
and activities, but none of these changes were comparable to the normalized gene
expression levels analysed for these stress proteins. GM maize seemed to induce
significant changes in white blood cell populations which are associated with an
immune response.

Publication Types:
Comparative Study

PMID: 17394522 [PubMed - indexed for MEDLINE]

688: Lipids. 2007 Apr;42(3):179-85. Epub 2007 Mar 14.

Engineering oilseed plants for a sustainable, land-based source of long chain
polyunsaturated fatty acids.

Damude HG, Kinney AJ.

Crop Genetics Research, DuPont Experimental Station, Wilmington, DE 19880-0353,
USA.

Numerous clinical studies have demonstrated the cardiovascular and mental health
benefits of including very long chain omega-3 polyunsaturated fatty acids, namely
eicospentaenoic acid (EPA) and docosohexaenoic acid (DHA) in the human diet.
Certain fish oils can be a rich source of omega-3 long chain polyunsaturated
fatty acids although processed marine oils are generally undesirable as food
ingredients because of the associated objectionable flavors and contaminants that
are difficult and cost-prohibitive to remove. Oilseed plants rich in omega-3
fatty acids, such as flax and walnut oils, contain only the 18-carbon omega-3
polyunsaturated fatty acid alpha-linolenic acid, which is poorly converted by the
human body to EPA and DHA. It is now possible to engineer common omega-6 rich
oilseeds such as soybean and canola to produce EPA and DHA and this has been the
focus of a number of academic and industrial research groups. Recent advances and
future prospects in the production of EPA and DHA in oilseed crops are discussed
here.

Publication Types:
Review

PMID: 17393224 [PubMed - indexed for MEDLINE]

689: Sci China C Life Sci. 2007 Feb;50(1):31-9.

Enhancing disease resistances of Super Hybrid Rice with four antifungal genes.

Zhu H, Xu X, Xiao G, Yuan L, Li B.

Biotechnology Research Center, Key Laboratory of Gene Engineering of Ministry of
Education, Sun Yat-sen University, Guangzhou, 510275, China.

A plant expression vector harboring four antifungal genes was delivered into the
embryogenic calli of '9311', an indica restorer line of Super Hybrid Rice, via
modified biolistic particle bombardment. Southern blot analysis indicated that in
the regenerated hygromycin-resistant plants, all the four antifungal genes,
including RCH10, RAC22, beta-Glu and B-RIP, were integrated into the genome of
'9311', co-transmitted altogether with the marker gene hpt in a Mendelian
pattern. Some transgenic R1 and R2 progenies, with all transgenes displaying a
normal expression level in the Northern blot analysis, showed high resistance to
Magnaporthe grisea when tested in the typical blast nurseries located in Yanxi
and Sanya respectively. Furthermore, transgenic F1 plants, resulting from a cross
of R2 homozygous lines with high resistance to rice blast with the non-transgenic
male sterile line Peiai 64S, showed not only high resistance to M. grisea but
also enhanced resistance to rice false smut (a disease caused by Ustilaginoidea
virens) and rice kernel smut (another disease caused by Tilletia barclayana).

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17393080 [PubMed - indexed for MEDLINE]

690: Asia Pac J Clin Nutr. 2007;16 Suppl 1:122-6.

Fruit quality of transgenic tomatoes with suppressed expression of LeETR1 and
LeETR2 genes.

Bao B, Ke L, Jiang J, Ying T.

Department of Food Science and Nutrition, Zhejiang University, 268 Kaixuan Road,
Hangzhou, Zhejiang, China 310029.

Tomato fruit is renowned for its high concentration of phyto-nutrients such as
lycopene and carotenoids, overall contribution to nutrition and human health. The
effect of antisense suppression of ethylene receptor genes LeETR1 and LeETR2 over
the quality of tomato fruit was investigated in this paper. During the different
stages of ripening, the fruit of antisense transgenic tomatoes of ale1 and ale2,
compared to their wild type B1, showed higher total soluble solids, acidity and
electrolytes accumulations and color development; lower fruit firmness, fruit
viscosity and fruit elasticity. However, no significant difference of Vc content,
total sugar, fruit pH value and fruit pigments between transgenic lines and B1
were noticed. ale1 and ale2 showed shortened shelf life. The data suggest that
fruit with suppressed LeETR1 and LeETR2 genes expression have stronger ethylene
response, which accelerate fruit ripening and greatly altered tomato variety
characteristics.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17392089 [PubMed - indexed for MEDLINE]

691: Br J Nutr. 2007 Jun;97(6):1083-9. Epub 2007 Mar 29.

L-arginine plus atorvastatin for prevention of atheroma formation in genetically
hypercholesterolaemic rabbits.

Rasmusen C, Moinard C, Martin C, Tricottet V, Cynober L, Couderc R.

Laboratory of Biological Nutrition EA 2498, Faculty of Pharmacy, Paris Descartes
University, 4 Avenue de l'Observatoire, 75270 Paris Cedex 06, France.

We investigated the combined effect of dietary supplementation with L-arginine,
which is the precursor of NO, and pharmacological treatment with atorvastatin,
which is a 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitor, on the
development of atherosclerosis in homozygous Watanabe heritable hyperlipidaemic
rabbits. Rabbits were fed either standard rabbit chow (group C; n 9) as control,
a 1.5 % L-arginine diet (group A; n 9), standard rabbit chow plus atorvastatin
(2.5 mg/kg per d) in drinking water (group S; n 8), or standard rabbit chow plus
a 1.5 % L-arginine diet with atorvastatin (group SA; n 8). Blood was sampled at
2-week intervals. After 8 weeks (T8), the aorta was harvested for topographic and
histological analysis. Only the SA group showed decreases in total area of
lesions (21 %) and the area of abdominal lesions (44 %) compared with the control
group (P = 0.019). Furthermore, plaques in the SA group were smaller and less
thick than those observed in the S group. Unexpectedly, plasma nitrite + nitrate
levels were not modified under either the L-arginine diet alone or under
L-arginine plus atorvastatin. The present study is the first to demonstrate that
diet supplementation with L-arginine associated with a statin (atorvastatin) is
more efficient in reducing lesion size than treatment with L-arginine or a statin
alone. This is a relatively novel therapeutic approach associating a
macronutrient and a drug.

PMID: 17391569 [PubMed - indexed for MEDLINE]

692: Plant Mol Biol. 2007 Jul;64(4):361-9. Epub 2007 Mar 28.

Transgenic Indian mustard (Brassica juncea) plants expressing an Arabidopsis
phytochelatin synthase (AtPCS1) exhibit enhanced As and Cd tolerance.

Gasic K, Korban SS.

Department of Natural Resources and Environmental Sciences, University of
Illinois at Urbana-Champaign, 310 ERML, 1201 W. Gregory Dr., Urbana, IL 61801,
USA.

Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides
that play important roles in detoxification of heavy metal and metalloids in
plants and other living organisms. The overall goal of this study is to develop
transgenic plants with increased tolerance for and accumulation of heavy metals
and metalloids from soil by expressing an Arabidopsis thaliana AtPCS1 gene,
encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A
FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian
mustard, and transgenic pcs lines have been compared with wild-type plants for
tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild
type plants, transgenic plants exhibit significantly higher tolerance to Cd and
As. Shoots of Cd-treated pcs plants have significantly higher concentrations of
PCs and thiols than those of wild-type plants. Shoots of wild-type plants
accumulated significantly more Cd than those of transgenic plants, while
accumulation of As in transgenic plants was similar to that in wild type plants.
Although phytochelatin synthase improves the ability of Indian mustard to
tolerate higher levels of the heavy metal Cd and the metalloid As, it does not
increase the accumulation potential of these metals in the above ground tissues
of Indian mustard plants.

PMID: 17390107 [PubMed - indexed for MEDLINE]

693: Am J Physiol Endocrinol Metab. 2007 Jul;293(1):E252-8. Epub 2007 Mar 27.

Melanocortin activation of nucleus of the solitary tract avoids anorectic
tachyphylaxis and induces prolonged weight loss.

Li G, Zhang Y, Rodrigues E, Zheng D, Matheny M, Cheng KY, Scarpace PJ.

Department of Pharmacology and Therapeutics, University of Florida College of
Medicine, Gainesville, Florida 32610, USA.

To examine the role of the brain stem melanocortin system in long-term energy
regulation, we assessed the effects of overproduction of proopiomelanocortin
(POMC) in the caudal brain stem of F344xBN rats with adult-onset obesity.
Recombinant adeno-associated viral vector encoding POMC gene was delivered to the
nucleus of solitary tract (NTS) in the hindbrain, and food intake, body weight,
glucose and fat metabolism, brown adipose tissue thermogenesis, and mRNA levels
of neuropeptides and melanocortin receptors were assessed. POMC delivery resulted
in sustained reduction in food intake and body weight over 42 days and improved
insulin sensitivity. At death, in recombinant adeno-associated viral
vector-POMC-treated rats vs. control rats, alpha-melanocyte-stimulating hormone
in NTS increased nearly 21-fold, whereas hypothalamic
alpha-melanocyte-stimulating hormone remained unchanged. Visceral adiposity
decreased by 37%; tissue triglyceride content diminished by 26% and 47% in liver
and muscle, respectively; serum triglyceride and nonesterified fatty acids were
reduced by 35% and 34%, respectively; phosphorylation of acetyl-CoA carboxylase
was elevated by 63% in soleus muscle; brown adipose tissue uncoupling protein 1
increased by 30%; and melanocortin 3 receptor expression declined by 60%, whereas
neuropeptide Y, agouti-related protein, and MC4 receptor mRNA levels were
unchanged in the NTS. In conclusion, POMC overexpression in the NTS produces a
characteristic unabated hypophagia that is uniquely different from the anorexic
tachyphylaxis following POMC overexpression in the hypothalamus. The sustained
anorectic response may result from absence of compensatory elements in the NTS,
such as increased agouti-related protein expression, suggesting melanocortin
activation of the brain stem may be a viable strategy to alleviate obesity.

Publication Types:
Evaluation Studies
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17389713 [PubMed - indexed for MEDLINE]

694: Anal Chim Acta. 2007 Feb 19;584(2):379-84. Epub 2006 Dec 3.

Discrimination of transgenic tomatoes based on visible/near-infrared spectra.

Xie L, Ying Y, Ying T, Yu H, Fu X.

College of Biosystems Engineering and Food Science, Zhejiang University, 268
Kaixuan St., 310029 Hangzhou, PR China.

VIS-NIR spectroscopy combined with multivariate analysis after the appropriate
spectral data pre-treatment has been proved to be a very powerful tool for
judgment of the relative pattern of the objects that have very similar
properties. In this study, seventy transgenic tomatoes with antisense LeETR2 and
94 of their parents, non-transgenic ones were measured in VIS-NIR diffuse
reflectance mode. Principal component analysis (PCA), discriminant analysis (DA)
and partial least-squares discriminant analysis (PLSDA) were applied to classify
tomatoes with different genes into two groups. Calibrations were developed using
PLS regression with the leave-one-out cross-validation technique. The results
show that differences between transgenic and non-transgenic tomatoes do exist and
excellent classification can be obtained after optimizing spectral pre-treatment.
The correct classifications for transgenic and non-transgenic tomatoes were both
100% using PLSDA after derivative spectral pre-treatment. The raw spectra with
PLSDA model after the second derivative pre-treatment had the best satisfactory
calibration and prediction abilities, with r(c)=0.97964, root mean square error
of calibration (RMSEC)=0.099, r(cv)=0.97963, root mean square error of
cross-validation (RMSECV)=0.0993 and a factor. The results in the present study
show VIS-NIR spectroscopy together with chemometrics techniques could be used to
differentiate transgenic tomato, which offers the benefit of avoiding
time-consuming, costly and laborious chemical and sensory analysis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17386628 [PubMed - indexed for MEDLINE]

695: Anal Chim Acta. 2007 Jan 2;581(1):78-82. Epub 2006 Aug 12.

Application of pressurized fluid extraction to determine cadmium and zinc in
plants.

Maurö­-Aucejo AR, Arnandis-Chover T, Marö­n-Sö¡ez R, Llobat-Estellö©s M.

Department of Analytical Chemistry, Universitat de Valö¨ncia, Dr. Moliner, 50,
46100 Burjassot, Valencia, Spain.

A procedure for the determination of Cd and Zn in plants is proposed. The metals
are extracted by pressurized fluid extraction (PFE). Operational conditions are:
pressure 1500 psi, temperature 75 degrees C, static time 5 min, flush volume 35%,
purge time 60s, cycles 1 and 1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid
(CDTA) 0.01M at pH 4.5 as extracting solution. Determination of Zn is carried out
by flame atomic absorption spectroscopy and depending on the concentration level,
Cd content is determined by flame or electrothermal atomic absorption
spectroscopy. Certified samples of Virginia tobacco leaves, tea leaves, spinach
leaves, poplar leaves, a commercial spinach sample (Spinacea oleracea) and
genetically modified Arabidopsis thaliana were analysed by the proposed procedure
and also by microwave acid digestion and extraction with HCl-Triton X-100.
Confidence intervals for Cd and Zn content obtained by the proposed procedure
overlap with the certified values. The other procedures, however, provide
inaccurate results for Cd. Recoveries obtained for a confidence level of 95% are
96+/-6% and 95+/-5% for Zn and Cd, respectively. Reproducibility of Zn by the
proposed procedure is 7% (n=8), similar to the other tests and the detection
limit is 2.6 microg. For Cd reproducibility is 8.5% (n=8), better than with
HCl-Triton X-100 and similar to acid digestion, the detection limit is 3.5 ng of
Cd.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 17386428 [PubMed - indexed for MEDLINE]

696: J Agric Food Chem. 2007 Apr 18;55(8):2998-3003. Epub 2007 Mar 27.

Expression of a maize Myb transcription factor driven by a putative silk-specific
promoter significantly enhances resistance to Helicoverpa zea in transgenic
maize.

Johnson ET, Berhow MA, Dowd PF.

Crop Bioprotection Research Unit, and New Crops and Processing Technology
Research, National Center for Agricultural Utilization Research, Agricultural
Research Service, U.S. Department of Agriculture, Peoria, Illinois 61604, USA.
johnet@ncaur.usda.gov

Hi II maize (Zea mays) plants were engineered to express maize p1 cDNA, a Myb
transcription factor, controlled by a putative silk specific promoter, for
secondary metabolite production and corn earworm resistance. Transgene expression
did not enhance silk color, but about half of the transformed plant silks
displayed browning when cut, which indicated the presence of p1-produced
secondary metabolites. Levels of maysin, a secondary metabolite with insect
toxicity, were highest in newly emerged browning silks. The insect resistance of
transgenic silks was also highest at emergence, regardless of maysin levels,
which suggests that other unidentified p1-induced molecules likely contributed to
larval mortality. Mean survivor weights of corn earworm larvae fed mature
browning transgenic silks were significantly lower than weights of those fed
mature nonbrowning transgenic silks. Some transgenic pericarps browned with
drying and contained similar molecules found in pericarps expressing a dominant
p1 allele, suggesting that the promoter may not be silk-specific.

PMID: 17385885 [PubMed - indexed for MEDLINE]

697: Arch Virol. 2007;152(7):1273-82. Epub 2007 Mar 26.

Suppressor of RNA silencing encoded by the monopartite tomato leaf curl Java
begomovirus.

Kon T, Sharma P, Ikegami M.

Department of Life Science, Graduate School of Agricultural Science, Tohoku
University, Sendai, Japan.

We previously isolated the monopartite begomovirus tomato leaf curl Java virus
(ToLCJAV) and satellite DNAbeta02 from the same naturally infected tomato source
in Indonesia. ToLCJAV induced mild leaf curl symptoms in Nicotiana benthamiana
plants; DNAbeta02 encoded the betaC1 gene and produced severe leaf curl symptoms
when co-inoculated with ToLCJAV in N. benthamiana. However, DNAbeta02mbetaC1,
which contains a frame shift mutation, did not induce severe symptoms in N.
benthamiana when co-inoculated with ToLCJAV. Expression of the betaC1 gene in N.
benthamiana using a potato virus X (PVX) vector induced virus-like symptoms in
the absence of ToLCJAV infection. When betaC1 and green fluorescent protein (GFP)
genes were co-expressed in the GFP-expressing N. benthamiana line 16c from a PVX
vector, betaC1 was able to suppress posttranscriptional gene silencing (PTGS)
induced by GFP and eliminated the short interfering RNA (siRNA) associated with
GFP expression, with a correlated increase in GFP mRNA accumulation. When C2 or
C4 genes of ToLCJAV and the GFP gene were co-expressed in the GFP-expressing N.
benthamiana line 16c, C2 showed a weak suppressor activity and C4 was unable to
suppress PTGS induced by GFP, and siRNA associated with GFP was detected. The
results of the sub-cellular localization of ToLCJAV-betaC1 in the epidermal cells
of N. benthamiana and onion tissues showed that this protein is accumulated
towards the periphery of the cell.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17385070 [PubMed - indexed for MEDLINE]

698: Plant Physiol. 2007 May;144(1):455-67. Epub 2007 Mar 23.

Cloning and characterization of unusual fatty acid desaturases from Anemone
leveillei: identification of an acyl-coenzyme A C20 Delta5-desaturase responsible
for the synthesis of sciadonic acid.

Sayanova O, Haslam R, Venegas Caleron M, Napier JA.

Rothamsted Research, Harpenden, Herts, United Kingdom. olga.sayanova@bbsrc.ac.uk

The seed oil of Anemone leveillei contains significant amounts of sciadonic acid
(20:3Delta(5,11,14); SA), an unusual non-methylene-interrupted fatty acid with
pharmaceutical potential similar to arachidonic acid. Two candidate cDNAs (AL10
and AL21) for the C(20) Delta(5cis)-desaturase from developing seeds of A.
leveillei were functionally characterized in transgenic Arabidopsis (Arabidopsis
thaliana) plants. The open reading frames of both Delta(5)-desaturases showed
some similarity to presumptive acyl-coenzyme A (CoA) desaturases found in animals
and plants. When expressed in transgenic Arabidopsis, AL21 showed a broad range
of substrate specificity, utilizing both saturated (16:0 and 18:0) and
unsaturated (18:2, n-6 and 18:3, n-3) substrates. In contrast, AL10 did not show
any activity in wild-type Arabidopsis. Coexpression of AL10 or AL21 with a C(18)
Delta(9)-elongase in transgenic Arabidopsis plants resulted in the production of
SA and juniperonic fatty acid (20:4Delta(5,11,14,17)). Thus, AL10 acted only on
C(20) polyunsaturated fatty acids in a manner analogous to "front-end"
desaturases. However, neither AL10 nor AL21 contain the cytochrome b(5) domain
normally present in this class of enzymes. Acyl-CoA profiling of transgenic
Arabidopsis plants and developing A. leveillei seeds revealed significant
accumulation of Delta(5)-unsaturated fatty acids as acyl-CoAs compared to the
accumulation of these fatty acids in total lipids. Positional analysis of
triacylglycerols of A. leveillei seeds showed that Delta(5)-desaturated fatty
acids were present in both sn-2 and sn-1 + sn-3 positions, although the majority
of 16:1Delta(5), 18:1Delta(5), and SA was present at the sn-2 position. Our data
provide biochemical evidence for the A. leveillei Delta(5)-desaturases using
acyl-CoA substrates.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17384161 [PubMed - indexed for MEDLINE]

699: Appetite. 2007 Jul;49(1):1-17. Epub 2007 Feb 24.

Consumer acceptance of technology-based food innovations: lessons for the future
of nutrigenomics.

Ronteltap A, van Trijp JC, Renes RJ, Frewer LJ.

Marketing and Consumer Behaviour Group, Wageningen University and Research
Centre, Hollandseweg 1, 6706 KN Wageningen, The Netherlands.
amber.ronteltap@wur.nl

Determinants of consumer adoption of innovations have been studied from different
angles and from the perspectives of various disciplines. In the food area, the
literature is dominated by a focus on consumer concern. This paper reviews
previous research into acceptance of technology-based innovation from both inside
and outside the food domain, extracts key learnings from this literature and
integrates them into a new conceptual framework for consumer acceptance of
technology-based food innovations. The framework distinguishes 'distal' and
'proximal' determinants of acceptance. Distal factors (characteristics of the
innovation, the consumer and the social system) influence consumers' intention to
accept an innovation through proximal factors (perceived cost/benefit
considerations, perceptions of risk and uncertainty, social norm and perceived
behavioural control). The framework's application as a tool to anticipate
consumer reaction to future innovations is illustrated for an actual
technology-based innovation in food science, nutrigenomics (the interaction
between nutrition and human genetics).

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17382433 [PubMed - indexed for MEDLINE]

700: FEMS Microbiol Ecol. 2007 Mar;59(3):600-10.

Bacterial community structures in honeybee intestines and their response to two
insecticidal proteins.

Babendreier D, Joller D, Romeis J, Bigler F, Widmer F.

Agroscope Reckenholz-Tö¤nikon Research Station ART, Zö¼rich, Switzerland.
dirk.babendreir@art.admin.ch

In this study, the effects of the Bt-toxin Cry1Ab and a soybean trypsin inhibitor
(SBTI) on intestinal bacterial communities of adult honeybees (Apis mellifera)
were investigated. It was hypothesized that changes in intestinal bacterial
communities of honeybees may represent a sensitive indicator for altered
intestinal physiology. Honeybees were fed in a laboratory set-up with maize
pollen from the Bt-transgenic cultivar MON810 or from the non-transgenic near
isoline. Purified Cry1Ab (0.0014% w/v) and SBTI (0.1% or 1% w/v) represented
supplementary treatments. For comparison, free-flying honeybees from two
locations in Switzerland were analysed. PCR-amplification of bacterial 16S rRNA
gene fragments and terminal restriction fragment length polymorphism analyses
revealed a total of 17 distinct terminal restriction fragments (T-RFs), which
were highly consistent between laboratory-reared and free-flying honeybees. The
T-RFs were affiliated to Alpha-, Beta-, and Gammaproteobacteria, to Firmicutes,
and to Bacteriodetes. Neither Bt-maize pollen nor high concentrations of Cry1Ab
significantly affected bacterial communities in honeybee intestines. Only the
high concentration of SBTI significantly reduced the number of T-RFs detected in
honeybee midguts, a concentration that also increases bee mortality. Therefore,
total bacterial community structures may not be a sensitive indicator for
providing evidence for the impact of insecticidal proteins on honeybees at
sublethal levels.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17381517 [PubMed - indexed for MEDLINE]

701: Biotechnol Lett. 2007 Jul;29(7):1135-42. Epub 2007 Mar 23.

Expression of alternansucrase in potato plants.

Kok-Jacon GA, Vincken JP, Suurs LC, Wang D, Liu S, Visser RG.

Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding,
Wageningen University, Wageningen, The Netherlands.

Alternan, which consists of alternating alpha-(1-->3)/alpha-(1-->6)-linked
glucosyl residues, was produced in potato tubers by expressing a mature
alternansucrase (Asr) gene from Leuconostoc mesenteroides NRRL B-1355 in potato.
Detection of alternan was performed by enzyme-linked immunosorbent assay in tuber
juices, revealing a concentration between 0.3 and 1.2 mg g(-1) fresh wt. The Asr
transcript levels correlated well with alternan accumulation in tuber juices. It
appeared that the expression of sucrose-regulated starch-synthesizing genes
(ADP-glucose pyrophosphorylase subunit S and granule-bound starch synthase I) was
down-regulated. Despite this, the physico-chemical properties of the transgenic
starches were unaltered. These results are compared to those obtained with other
transgenic potato plants producing mutan [alpha-(1-->3)-linked glucosyl residues]
and dextran [alpha-(1-->6)-linked glucosyl residues].

PMID: 17380272 [PubMed - indexed for MEDLINE]

702: Plant Cell Physiol. 2007 Apr;48(4):626-37. Epub 2007 Mar 22.

Increased Rubisco content in transgenic rice transformed with the 'sense' rbcS
gene.

Suzuki Y, Ohkubo M, Hatakeyama H, Ohashi K, Yoshizawa R, Kojima S, Hayakawa T,
Yamaya T, Mae T, Makino A.

Department of Applied Plant Science, Graduate School of Agricultural Science,
Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Sendai 981-8555, Japan.

Rice (Oryza sativa L.) plants with substantially increased Rubisco content were
obtained by Agrobacterium-mediated transformation with the rice rbcS sense gene
under the control of the rice rbcS promoter. The primary transformants were
screened for the ratio of Rubisco to leaf-N content, and the transformants with
>120% wild-type levels of Rubisco were selected. In the progeny of the selected
lines of the transformants, the mRNA levels of one member of the rbcS gene family
were increased from 3.9- to 6.2-fold, whereas those of other members of the rbcS
gene family were unchanged. The total levels of rbcS mRNA were increased from
2.1- to 2.8-fold. The levels of rbcL mRNA were increased from 1.2- to 1.9-fold.
Rubisco protein content was significantly increased by 30% on a leaf area basis.
The ratio of Rubisco-N to leaf-N was also increased by 10-20%, irrespective of N
treatment. The specific activity of Rubisco per unit of enzyme protein was not
different. However, light-saturated photosynthesis was not enhanced even when the
rate was measured at low [CO2] where Rubisco becomes limiting for photosynthesis.
Some lines showed lower photosynthesis at high [CO2] (>60 Pa). We conclude that
introduction of additional sense rbcS leads to overexpression of rbcS and that
this overexpression slightly up-regulates the gene expression of rbcL at the
transcript level and enhances the amount of Rubisco holoenzyme. However,
overproduction of Rubisco protein does not improve photosynthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17379698 [PubMed - indexed for MEDLINE]

703: Mol Plant Microbe Interact. 2007 Mar;20(3):276-82.

Coil-dependent signaling pathway is not required for Mi-1-mediated potato aphid
resistance.

Bhattarai KK, Xie QG, Pourshalimi D, Younglove T, Kaloshian I.

Department of Nematology, University of California, Riverside, CA 92521, USA.

Tomato (Solanum lycopersicum) has a unique resistance gene, Mi-1, that confers
resistance to animals from distinct taxa, nematodes, and piercing and sucking
insects. Mi-1 encodes a protein with a nucleotide-binding site and leucine-rich
repeat motifs. Early in the potato aphid (Macrosiphum euphorbiae)--tomato
interactions, aphid feeding induces the expression of the jasmonic acid
(JA)-regulated proteinase inhibitor genes, Pin1 and Pin2. The jail-1 (jasmonic
acid insensitive 1) tomato mutant, which is impaired in JA perception, was used
to gain additional insight into the JA signaling pathway and its role in the
Mi-1-mediated aphid resistance. The jail-1 mutant has a deletion in the Coil gene
that encodes a putative F-box protein. In this study, aphid colonization,
survival, and fecundity were compared on wild-type tomato and jail-1 mutant. In
choice assays, the jail-1 mutant showed higher colonization by potato aphids
compared with wild-type tomato. In contrast, no-choice assays showed no
difference in potato aphid survival or fecundity between jail-1 and the wild-type
parent. Plants homozygous for Mi-1 and for the jail mutation were not compromised
in resistance to potato aphids, using either choice or no-choice assays. In
addition, the accumulation of JA-regulated Pin1 transcripts after aphid feeding
was Coil dependent. Taken together, these data indicate that, although potato
aphids activate Coil-dependent defense response in tomato, this response is not
required for Mi-1-mediated resistance to aphids.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17378430 [PubMed - indexed for MEDLINE]

704: Plant Cell Rep. 2007 Aug;26(8):1221-31. Epub 2007 Mar 22.

Variable T-DNA linkage configuration affects inheritance of carotenogenic
transgenes and carotenoid accumulation in transgenic indica rice.

Rai M, Datta K, Parkhi V, Tan J, Oliva N, Chawla HS, Datta SK.

Plant Breeding, Genetics and Biotechnology Division, International Rice Research
Institute, DAPO Box 7777, Metro Manila, Philippines. mynkrai@yahoo.co.in

Transgenics for the expression of beta-carotene biosynthetic pathway in the
endosperm were developed in indica rice background by introducing phytoene
synthase (psy) and phytoene desaturase (crtI) genes through
Agrobacterium-mediated transformation, employing non-antibiotic positive
selectable marker phosphomannose isomerase (pmi). Twenty-seven transgenic lines
were characterized for the structural organization of T-DNA inserts and the
expression of transgenes in terms of total carotenoid and beta-carotene
accumulation in the endosperm. Ten lines were also studied for the inheritance of
transgenic loci to the T(1) progenies. Copy number and sites of integration of
the transgenes ranged from one to four. Almost 50% of the transgenic lines showed
rearrangement of T-DNA inserts. However, most of the rearrangements occurred in
the crtI expression cassette which is adjacent to the right T-DNA border.
Differences in copy numbers of psy and crtI were also observed indicating partial
T-DNA integration. Beyond T-DNA border transfer was also detected in 25% of the
lines. Fifty percent of the lines studied showed single Mendelian locus
inheritance, while two lines showed bi-locus inheritance in the T(1) progenies.
Some of the lines segregating in 3:1 ratio showed two sites of integration on
restriction digestion analysis indicating that the T-DNA insertion sites were
tightly linked. Three transgenic lines showed nonparental types in the
segregating progenies, indicating unstable transgenic locus. Evidences from the
HPLC analysis showed that multiple copies of transgenes had a cumulative effect
on the accumulation of carotenoid in the endosperm. T(1) progenies, in general,
accumulated more carotenoids than their respective parents, the highest being
6.77 mug/g of polished seeds. High variation in the carotenoid accumulation was
observed within the T(1) progenies which could be attributed to the variation in
the structural organization and expression of transgenes, minor variations in the
genetic background within the progeny plants, or differences in the plant
microenvironments. The study identified lines worthy of further multiplication
and breeding based on transgene structural integrity in the segregating progeny
and high expression levels in terms of the beta-carotene accumulation.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17377795 [PubMed - indexed for MEDLINE]

705: Plant J. 2007 Apr;50(2):364-79. Epub 2007 Mar 21.

Further in vivo studies on the role of the molecular chaperone, Hsp93, in plastid
protein import.

Kovacheva S, Bö©dard J, Wardle A, Patel R, Jarvis P.

Department of Biology, University of Leicester, Leicester LE1 7RH, UK.

In Arabidopsis, Hsp93 is encoded by two genes, atHSP93-V and atHSP93-III. We
identified two T-DNA mutants for atHSP93-III: one being a partial 'knockdown'
(hsp93-III-1) and the other a complete 'knockout' (hsp93-III-2). Homozygotes for
both mutants were indistinguishable from wild type. We crossed each mutant to an
atHSP93-V knockout, and identified double mutants with strongly chlorotic
phenotypes. This implied redundancy, which was confirmed by the complementation
of mildly chlorotic hsp93-V plants by atHSP93-III over-expression. While the
hsp93-V hsp93-III-1 mutant was doubly homozygous, the second double mutant was
heterozygous for hsp93-III-2 (genotype: hsp93-V/hsp93-V; +/hsp93-III-2). Attempts
to identify an hsp93-V hsp93-III-2 double homozygote were unsuccessful,
indicating that the Hsp93 pool is essential for viability. Consistently, siliques
of the second double mutant contained aborted seeds (because of a block in the
zygote-embryo transition) and failed ovules (because of a moderate defect in
female gametophytes). Double-mutant plants were chlorophyll-deficient, contained
under-developed chloroplasts, and exhibited stunted growth. In import assays
using a chimeric pre-protein (plastocyanin transit peptide fused to dihydrofolate
reductase; PC-DHFR), a clear defect was observed in hsp93-V hsp93-III-1
chloroplasts. Interestingly, while denaturation or stabilization of the DHFR
moiety had a strong effect on import efficiency in the wild type, no such effects
were observed with double-mutant (or tic40) chloroplasts. This indicated that
pre-protein unfolding is not rate-limiting for import into mutant chloroplasts,
and suggested that (unlike the situation in mitochondria) the inner membrane
import machinery does not contribute to pre-protein unfolding at the organellar
surface.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17376159 [PubMed - indexed for MEDLINE]

706: Trends Biotechnol. 2007 May;25(5):201-3. Epub 2007 Mar 19.

Food from cloned animals is part of our brave old world.

Miller HI.

The Hoover Institution, Stanford University, Stanford, CA 94305-6010, USA.
miller@hoover.stanford.edu

When confronted by pressure from activists and Congress, the US Food and Drug
Administration (FDA) has not always adopted policies and made decisions about
individual products that accord with the scientific evidence. An example was the
unnecessarily and markedly prolonged review of the veterinary drug bovine
somatotropin (bST), or bovine growth hormone, during the 1980s. The FDA now faces
a similar situation surrounding the question of whether meat and milk from cloned
animals and their offspring are safe for human consumption. Having made a
preliminary decision in the affirmative - based on an exhaustive analysis of
scientific articles, health records, blood samples and studies of the composition
of meat and milk - the agency has been beleaguered by criticisms. It remains to
be seen whether, ultimately, science will trump anti-technology, anti-consumer
activism.

Publication Types:
Review

PMID: 17374411 [PubMed - indexed for MEDLINE]

707: Proc Natl Acad Sci U S A. 2007 Mar 27;104(13):5580-3. Epub 2007 Mar 19.

Transgenic malaria-resistant mosquitoes have a fitness advantage when feeding on
Plasmodium-infected blood.

Marrelli MT, Li C, Rasgon JL, Jacobs-Lorena M.

Department of Molecular Microbiology and Immunology, Bloomberg School of Public
Health and Malaria Research Institute, The Johns Hopkins University, Baltimore,
MD 21205, USA.

The introduction of genes that impair Plasmodium development into mosquito
populations is a strategy being considered for malaria control. The effect of the
transgene on mosquito fitness is a crucial parameter influencing the success of
this approach. We have previously shown that anopheline mosquitoes expressing the
SM1 peptide in the midgut lumen are impaired for transmission of Plasmodium
berghei. Moreover, the transgenic mosquitoes had no noticeable fitness load
compared with nontransgenic mosquitoes when fed on noninfected mice. Here we show
that when fed on mice infected with P. berghei, these transgenic mosquitoes are
more fit (higher fecundity and lower mortality) than sibling nontransgenic
mosquitoes. In cage experiments, transgenic mosquitoes gradually replaced
nontransgenics when mosquitoes were maintained on mice infected with
gametocyte-producing parasites (strain ANKA 2.34) but not when maintained on mice
infected with gametocyte-deficient parasites (strain ANKA 2.33). These findings
suggest that when feeding on Plasmodium-infected blood, transgenic
malaria-resistant mosquitoes have a selective advantage over nontransgenic
mosquitoes. This fitness advantage has important implications for devising
malaria control strategies by means of genetic modification of mosquitoes.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 17372227 [PubMed - indexed for MEDLINE]

708: Magn Reson Imaging. 2007 Apr;25(3):425-32. Epub 2006 Nov 20.

Water mobility in the endosperm of high beta-glucan barley mutants as studied by
nuclear magnetic resonance imaging.

Fast Seefeldt H, van den Berg F, Kö¶ckenberger W, Engelsen SB, Wollenweber B.

Department of Genetics and Biotechnology, Danish Institute of Agricultural
Sciences, Research Centre Flakkebjerg, DK-4200 Slagelse, Denmark.
helenef.seefeldt@agrsci.dk

(1)H NMR imaging (MRI) was used as a noninvasive technique to study water
distribution and mobility in hydrated barley (Hordeum vulgare L.) seeds of
accessions with varying content of beta glucan (BG), a highly hygroscopic cell
wall component. High contents of BG in barley are unfavorable in malting where it
leads to clotting of filters and hazing of beer as well as in animal feed where
it hinders the rapid uptake of energy. However, a high content of BG has a
positive nutritional effect, as it lowers the cholesterol and the glycaemic
index. It was studied whether water distribution and mobility were related to
content and location of BG. Water mobility was investigated by following the rate
and mode of desiccation in hydrated single seeds. In order to determine the
different water components, a multispin echo experiment was set up to reveal the
T(2) transverse relaxation rates of water within the seeds. A principal component
analysis (PCA) discriminated control seeds from the high-BG mutant seeds. MRI
proved efficient in tracing the differences in water-holding capacity of
contrasting barley seeds. All accessions showed nonuniform distribution of water
at full hydration as well as during desiccation. The embryo retained water even
after 36 h of drying, whereas the endosperm showed low and heterogeneous mobility
of the water after drying. The relaxation time constants indicated that the BG
mutants had regions of much higher water mobility around the ventral crease
compared to the control. It is concluded that MRI can be applied to investigate
temporal and spatial differences in the location of specific chemical compounds
in single seeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17371735 [PubMed - indexed for MEDLINE]

709: J Agric Food Chem. 2007 Apr 18;55(8):2918-22. Epub 2007 Mar 20.

Qualitative and quantitative polymerase chain reaction assays for an alfalfa
(Medicago sativa)-specific reference gene to use in monitoring transgenic
cultivars.

Alexander TW, Reuter T, McAllister TA.

Agriculture and Agri-Food Canada Research Centre, P.O. Box 3000, Lethbridge,
Alberta, Canada.

Genetically modified (GM) alfalfa (Medicago sativa) was marketed for the first
time in 2005. For countries with established thresholds for GM plants, methods to
detect and quantify their adventitious presence are required. We selected acetyl
CoA carboxylase as a reference gene for the detection and quantification of GM
alfalfa. Two qualitative polymerase chain reaction (PCR) assays (Acc1 and Acc2)
were designed to detect alfalfa. Both were specific to alfalfa, amplifying DNA
from 12 separate cultivars and showing negative results for PCR of 15 nonalfalfa
plants. The limits of detection for Acc1 and Acc2 were 0.2 and 0.01%,
respectively. A quantitative real-time PCR assay was also designed, having high
linearity (r > 0.99) over alfalfa standard concentrations ranging from 100 to 2.0
x 10(5) pg of alfalfa DNA per PCR. The real-time PCR assay was effective in
quantifying alfalfa DNA from forage- and concentrate-based mixed diets containing
different amounts of alfalfa meal.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17371040 [PubMed - indexed for MEDLINE]

710: J Econ Entomol. 2007 Feb;100(1):172-9.

Resistance to Tecia solanivora (Lepidoptera: Gelechiidae) in three transgenic
Andean varieties of potato expressing Bacillus thuringiensis CrylAc protein.

Valderrama AM, Veö¡squez N, Rodrö­guez E, Zapata A, Zaidi MA, Altosaar I, Arango R.

Unidad de Biotecnologö­a Vegetal UNALMED-CIB, Corporaciö¨n para Investigaciones
Biolö¨gicas, (CIB), Medellin, Antioquia, Colombia.

Transgenic potato, Solanum tuberosum L., plants containing a synthetic cry1Ac
gene coding for the Bacillus thuringiensis (Bt) crystalline insecticidal protein
were produced and evaluated for resistance to Tecia solanivora Povolny
(Lepidoptera: Gelechiidae), the larvae of which attack potato tubers. In total,
43 transgenic lines of commercial Andean potato varieties Diacol Capiro, Pardo
Pastusa, and Pandeazöºcar were obtained. These transgenic lines were found to have
one to four copies of cry1Ac per genome and expression levels of Cry1Ac protein
varying from 0.02 to 17 microg/g fresh tuber tissue. Bioassays of T. solanivora
larvae on these transgenic potato tubers showed 83.7-100% mortality, whereas the
mortality levels on nontransgenic lines were 0-2.67%. Our data indicate the
capability of Bt transgenic technology to control the T. solanivora while
reducing the use of chemical insecticides. Further studies under controlled field
conditions will be helpful in exploring the potential of CrylAc potatoes in the
insect pest management strategies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17370825 [PubMed - indexed for MEDLINE]

711: J Econ Entomol. 2007 Feb;100(1):164-71.

Sugarcane borer (Lepidoptera: Crambidae) resistance to transgenic Bacillus
thuringiensis maize.

Huang F, Leonard BR, Andow DA.

Department of Entomology, Louisiana State University AgCenter, Baton Rouge, LA
70803, USA. fhuang@agcenter.lsu.edu

Transgenic maize, Zea mays L., expressing the Bacillus thuringiensis (Bt) CrylAb
toxin has been planted to extensive areas across the United States and several
other countries, but no resistance has been documented in field populations
oflepidopteran target pests. This article describes the first report of
resistance alleles to commercially available Cry1Ab Bt maize in a Louisiana
population of sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera:
Crambidae). Two hundred thirteen two-parent isolines of D. saccharalis were
screened for Cry1Ab resistance on Bt maize leaf tissue using an F2 screening
technique. Larvae representing three isolines survived >15 d on Bt tissue in the
F2 generation. The second generation backcross progeny (B1F2) derived from
isoline 52 completed larval development on Bt maize in the greenhouse.
Segregation and resistance frequency analysis associated with isoline 52
suggested that Bt resistance is probably determined by a nearly completely
recessive allele at a single locus. With this assumption, the estimated
resistance allele frequency in this population is 0.0023, within a 95% confidence
interval of 0.0003-0.0064.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17370824 [PubMed - indexed for MEDLINE]

712: Nat Genet. 2007 Apr;39(4):544-9. Epub 2007 Mar 18.

The heterochronic maize mutant Corngrass1 results from overexpression of a tandem
microRNA.

Chuck G, Cigan AM, Saeteurn K, Hake S.

Plant Gene Expression Center, 800 Buchanan St., Albany, California 94710, USA.
gchuck@nature.berkeley.edu

Retention of juvenile traits in the adult reproductive phase characterizes a
process known as neoteny, and speculation exists over whether it has contributed
to the evolution of new species. The dominant Corngrass1 (Cg1) mutant of maize is
a neotenic mutation that results in phenotypes that may be present in the
grass-like ancestors of maize. We cloned Cg1 and found that it encodes two tandem
miR156 genes that are overexpressed in the meristem and lateral organs.
Furthermore, a target of Cg1 is teosinte glume architecture1 (tga1), a gene known
to have had a role in the domestication of maize from teosinte. Cg1 mutant plants
overexpressing miR156 have lower levels of mir172, a microRNA that targets genes
controlling juvenile development. By altering the relative levels of both
microRNAs, it is possible to either prolong or shorten juvenile development in
maize, thus providing a mechanism for how species-level heterochronic changes can
occur in nature.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17369828 [PubMed - indexed for MEDLINE]

713: Plant Physiol. 2007 May;144(1):121-33. Epub 2007 Mar 16.

The rice YABBY1 gene is involved in the feedback regulation of gibberellin
metabolism.

Dai M, Zhao Y, Ma Q, Hu Y, Hedden P, Zhang Q, Zhou DX.

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural
University, Wuhan 430070, China.

Gibberellin (GA) biosynthesis is regulated by feedback control providing a
mechanism for GA homeostasis in plants. However, regulatory elements involved in
the feedback control are not known. In this report, we show that a rice (Oryza
sativa) YABBY1 (YAB1) gene had a similar expression pattern as key rice GA
biosynthetic genes GA3ox2 and GA20ox2. Overexpression of YAB1 in transgenic rice
resulted in a semidwarf phenotype that could be fully rescued by applied GA.
Quantification of the endogenous GA content revealed increases of GA(20) and
decreases of GA(1) levels in the overexpression plants, in which the transcripts
of the biosynthetic gene GA3ox2 were decreased. Cosuppression of YAB1 in
transgenic plants induced expression of GA3ox2. The repression of GA3ox2 could be
obtained upon treatment by dexamethasone of transgenic plants expressing a
YAB1-glucocorticoid receptor fusion. Importantly, we show that YAB1 bound to a
GA-responsive element within the GA3ox2 promoter. In addition, the expression of
YAB1 was deregulated in GA biosynthesis and signaling mutants and could be either
transiently induced by GA or repressed by a GA inhibitor. Finally, either
overexpression or cosuppression of YAB1 impaired GA-mediated repression of
GA3ox2. These data together suggest that YAB1 is involved in the feedback
regulation of GA biosynthesis in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17369428 [PubMed - indexed for MEDLINE]

714: DNA Seq. 2007 Apr;18(2):131-7.

Molecular cloning and characterization of ETHYLENE OVERPRODUCER 1-LIKE 1 gene,
LeEOL1, from tomato (Lycopersicon esculentum Mill.) fruit.

Zhu HL, Zhu BZ, Shao Y, Lin XJ, Wang XG, Gao HY, Xie YH, Li YC, Luo YB.

Laboratory of Fruit Biology, College of Food Science & Nutritional Engineering,
China Agricultural University, No. 17 Qinghua Donglu, Beijing 100083, People's
Republic of China.

Recently, ETHYLENE OVERPRODUCER 1 (ETO1) had been cloned and identified as a
negative post-transcriptional regulator in the ethylene biosynthesis in
Arabidopsis. However, little was known about the role of ETO1 in other species,
especially in tomato, which was an ideal model for studying the biosynthesis of
ethylene during tomato fruit ripening. In this study, a tomato ETHYLENE
OVERPRODUCER 1-LIKE 1 (LeEOL1) was cloned. The LeEOL1 cDNA was 3,515 bp long and
carried an ORF that putatively encoded a polypeptide of 886 amino acids with a
predicted molecular mass of 95 kDa. It shared 74% identity in amino acid sequence
with Arabidopsis EOL1 and had one BTB (Broad-complex, Tramtrack, Bric-ö -brac)
domain and two TPR (tetratricopeptide repeat) domains, which were also conserved
domains in AtEOL1. RT-PCR analysis of the temporal expression of LeEOL1 showed
that its transcript decreased companied with increase of ethylene production in
tomato ripening. The level of LeEOL1 transcripts in wild type tomato fruit at
mature green stage did not distinctively change when treated with exogenous
ethylene.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17364824 [PubMed - indexed for MEDLINE]

715: Plant Mol Biol. 2007 May;64(1-2):1-15. Epub 2007 Mar 16.

Overexpression of salicylic acid carboxyl methyltransferase reduces salicylic
acid-mediated pathogen resistance in Arabidopsis thaliana.

Koo YJ, Kim MA, Kim EH, Song JT, Jung C, Moon JK, Kim JH, Seo HS, Song SI, Kim
JK, Lee JS, Cheong JJ, Choi YD.

School of Agricultural Biotechnology, Seoul National University, Seoul 151-742,
Korea.

We cloned a salicylic acid/benzoic acid carboxyl methyltransferase gene, OsBSMT1,
from Oryza sativa. A recombinant OsBSMT1 protein obtained by expressing the gene
in Escherichia coli exhibited carboxyl methyltransferase activity in reactions
with salicylic acid (SA), benzoic acid (BA), and de-S-methyl
benzo(1,2,3)thiadiazole-7-carbothioic acid (dSM-BTH), producing methyl salicylate
(MeSA), methyl benzoate (MeBA), and methyl dSM-BTH (MeBTH), respectively.
Compared to wild-type plants, transgenic Arabidopsis overexpressing OsBSMT1
accumulated considerably higher levels of MeSA and MeBA, some of which were
vaporized into the environment. Upon infection with the bacterial pathogen
Pseudomonas syringae or the fungal pathogen Golovinomyces orontii, transgenic
plants failed to accumulate SA and its glucoside (SAG), becoming more susceptible
to disease than wild-type plants. OsBSMT1-overexpressing Arabidopsis showed
little induction of PR-1 when treated with SA or G. orontii. Notably, incubation
with the transgenic plant was sufficient to trigger PR-1 induction in neighboring
wild-type plants. Together, our results indicate that in the absence of SA, MeSA
alone cannot induce a defense response, yet it serves as an airborne signal for
plant-to-plant communication. We also found that jasmonic acid (JA) induced
AtBSMT1, which may contribute to an antagonistic effect on SA signaling pathways
by depleting the SA pool in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17364223 [PubMed - indexed for MEDLINE]

716: Plant Biotechnol J. 2007 May;5(3):422-30. Epub 2007 Mar 15.

Erratum in:
Plant Biotechnol J. 2007 Sep;5(5):675.. Corrected Jeknciä‡ to Jekniä‡..

The codA transgene for glycinebetaine synthesis increases the size of flowers and
fruits in tomato.

Park EJ, Jekniä‡ Z, Chen TH, Murata N.

Department of Horticulture, ALS4017, Oregon State University, Corvallis, OR
97331, USA.

The tolerance of various species of plant to abiotic stress has been enhanced by
genetic engineering with certain genes. However, the use of such transgenes is
often associated with negative effects on growth and productivity under
non-stress conditions. The codA gene from Arthrobacter globiformis is of
particular interest with respect to the engineering of desirable productive
traits in crop plants. The expression of this gene in tomato plants resulted in
significantly enlarged flowers and fruits under non-stress conditions. The
enlargement of flowers and fruits was associated with high levels of
glycinebetaine that accumulated in reproductive organs, such as flower buds and
fruits. The enlargement of flowers was related to an increase in the size and
number of cells, and reflected the pleiotropic effect of the codA transgene on
the expression of genes involved in the regulation of cell division.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17362485 [PubMed - indexed for MEDLINE]

717: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4218-22. Epub 2007 Mar 5.

Comment in:
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3675-6.

Folate biofortification of tomato fruit.

Dö­az de la Garza RI, Gregory JF 3rd, Hanson AD.

Department of Horticultural Sciences, University of Florida, Gainesville, FL
32611, USA.

Folate deficiency leads to neural tube defects and other human diseases, and is a
global health problem. Because plants are major folate sources for humans, we
have sought to enhance plant folate levels (biofortification). Folates are
synthesized from pteridine, p-aminobenzoate (PABA), and glutamate precursors.
Previously, we increased pteridine production in tomato fruit up to 140-fold by
overexpressing GTP cyclohydrolase I, the first enzyme of pteridine synthesis.
This strategy increased folate levels 2-fold, but engineered fruit were
PABA-depleted. We report here the engineering of fruit-specific overexpression of
aminodeoxychorismate synthase, which catalyzes the first step of PABA synthesis.
The resulting fruit contained an average of 19-fold more PABA than controls. When
transgenic PABA- and pteridine-overproduction traits were combined by crossing,
vine-ripened fruit accumulated up to 25-fold more folate than controls. Folate
accumulation was almost as high (up to 15-fold) in fruit harvested green and
ripened by ethylene-gassing, as occurs in commerce. The accumulated folates
showed normal proportions of one-carbon forms, with 5-methyltetrahydrofolate the
most abundant, but were less extensively polyglutamylated than controls. Folate
concentrations in developing fruit did not change in controls, but increased
continuously throughout ripening in transgenic fruit. Pteridine and PABA levels
in transgenic fruit were >20-fold higher than in controls, but the pathway
intermediates dihydropteroate and dihydrofolate did not accumulate, pointing to a
flux constraint at the dihydropteroate synthesis step. The folate levels we
achieved provide the complete adult daily requirement in less than one standard
serving.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17360503 [PubMed - indexed for MEDLINE]

718: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3675-6. Epub 2007 Mar 5.

Comment on:
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4218-22.

Biofortification of plant-based food: enhancing folate levels by metabolic
engineering.

DellaPenna D.

Department of Biochemistry and Molecular Biology, Michigan State University, East
Lansing, MI 48824-1319, USA. dellapen@msu.edu

Publication Types:
Comment

PMID: 17360411 [PubMed - indexed for MEDLINE]

719: Trends Plant Sci. 2007 Apr;12(4):177-83. Epub 2007 Mar 13.

Genetic use restriction technologies (GURTs): strategies to impede transgene
movement.

Hills MJ, Hall L, Arnison PG, Good AG.

Department of Biological Sciences, University of Alberta, Edmonton, AB, T6H 2X6,
Canada.

No clear consensus has emerged in the debate about the risks posed by transgenic
crops and how to assess these risks accurately. In the meantime, interest is
growing in strategies to impede transgene movement. This attention is being
driven, in part, by expanding interest in using transgenic crops to produce
pharmaceutical and industrial products. Potential strategies to impede transgene
movement have been published in the scientific literature, and numerous patents
have been submitted; however, the efficacy of such strategies has still to be
evaluated in a field situation. In this review, we discuss some of the genetic
strategies that could be used to restrict the spread of transgenes, although at
present many of these technologies are still largely at a theoretical stage of
development.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17360223 [PubMed - indexed for MEDLINE]

720: Plant Biotechnol J. 2007 May;5(3):381-8. Epub 2007 Mar 15.

Viability and bar expression are negatively correlated in Oregon Wolfe Barley
Dominant hybrids.

Bregitzer P, Cooper LD, Hayes PM, Lemaux PG, Singh J, Sturbaum AK.

National Small Grains Germplasm Research Facility, USDA-ARS, Aberdeen, ID 83210,
USA. pbregit@uidaho.edu

The expression level of bar, which encodes phosphinothricin acetyltransferase
(PAT), was correlated with the inviability of barley hybrids between 20 Golden
Promise-derived transgenic lines (Ds-bar lines) and a specialized genetic marker
stock, Oregon Wolfe Barley Dominant (OWBD). Each Ds-bar line was homozygous for a
modified maize Ds element that encoded bar and that had been delivered via
transposition to a unique location. All Ds-bar lines were viable and
morphologically similar. Only four of the 20 hybrid populations were viable. The
remaining populations died prior to producing seed. Phenotypic, enzyme-linked
immunosorbent assay and quantitative reverse transcriptase-polymerase chain
reaction analyses of these lines, and of lines from unrelated transformation
events that also expressed bar, showed that viability was negatively correlated
with bar expression. Analysis of crosses of a high-bar-expressing line with the
OWB mapping population showed that the sensitivity of OWBD to PAT segregated as a
single locus on chromosome 6HL. No sensitivity to PAT could be detected in
several other lines and cultivars. OWBD has been shown to be genetically
divergent from other germplasm groups within cultivated barley; therefore, the
observed sensitivity may be peculiar to OWBD and thus would not impact generally
on the utility of bar as a selectable marker or source of herbicide resistance in
barley. Nevertheless, these results demonstrate the extent of allelic variability
present in Hordeum vulgare, and suggest an additional variable for consideration
when devising protocols for the transformation of Hordeum cultivars or landraces
that are not known to be tolerant to PAT.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17359497 [PubMed - indexed for MEDLINE]

721: Arch Environ Contam Toxicol. 2007 May;52(4):596-602. Epub 2007 Mar 13.

New analysis of a rat feeding study with a genetically modified maize reveals
signs of hepatorenal toxicity.

Sö©ralini GE, Cellier D, de Vendomois JS.

Committee for Independent Information and Research on Genetic Engineering
CRIIGEN, Paris, France. criigen@unicaen.fr

Health risk assessment of genetically modified organisms (GMOs) cultivated for
food or feed is under debate throughout the world, and very little data have been
published on mid- or long-term toxicological studies with mammals. One of these
studies performed under the responsibility of Monsanto Company with a transgenic
corn MON863 has been subjected to questions from regulatory reviewers in Europe,
where it was finally approved in 2005. This necessitated a new assessment of
kidney pathological findings, and the results remained controversial. An Appeal
Court action in Germany (Mö¼nster) allowed public access in June 2005 to all the
crude data from this 90-day rat-feeding study. We independently re-analyzed these
data. Appropriate statistics were added, such as a multivariate analysis of the
growth curves, and for biochemical parameters comparisons between GMO-treated
rats and the controls fed with an equivalent normal diet, and separately with six
reference diets with different compositions. We observed that after the
consumption of MON863, rats showed slight but dose-related significant variations
in growth for both sexes, resulting in 3.3% decrease in weight for males and 3.7%
increase for females. Chemistry measurements reveal signs of hepatorenal
toxicity, marked also by differential sensitivities in males and females.
Triglycerides increased by 24-40% in females (either at week 14, dose 11% or at
week 5, dose 33%, respectively); urine phosphorus and sodium excretions
diminished in males by 31-35% (week 14, dose 33%) for the most important results
significantly linked to the treatment in comparison to seven diets tested. Longer
experiments are essential in order to indicate the real nature and extent of the
possible pathology; with the present data it cannot be concluded that GM corn
MON863 is a safe product.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17356802 [PubMed - indexed for MEDLINE]

722: Curr Opin Biotechnol. 2007 Apr;18(2):115-20. Epub 2007 Mar 13.

GM maize from site-specific recombination technology, what next?

Ow DW.

Plant Gene Expression Center, USDA-ARS and University of California at Berkeley,
800 Buchanan Street, Albany, California 94710, USA. david_ow@berkeley.edu

The term plant genetic engineering has long conveyed a highly efficient and
precise process for the manipulation of plant genomes. For nearly two decades,
research on recombinase-based applications has steadily advanced the surgical
capabilities of plant genome rearrangements. Once considered interesting
laboratory exercises, a first crop plant derived from this type of DNA acrobatics
is heading to market. Originally configured for a specific application, to remove
a selectable marker, it could be the first of more to come - and not just
market-free plants.

Publication Types:
Review

PMID: 17353124 [PubMed - indexed for MEDLINE]

723: Peptides. 2007 May;28(5):974-80. Epub 2007 Feb 6.

Is the titer of adipokinetic peptides in Leptinotarsa decemlineata fed on
genetically modified potatoes increased by oxidative stress?

Kodrö­k D, Krishnan N, Habustovö¡ O.

Institute of Entomology, Biology Centre, Czech Academy of Sciences, Branisovskö¡
31, Ceskö© Budejovice, Czech Republic. kodrik@entu.cas.cz

The level of adipokinetic hormones (AKHs) (Peram-CAH-I and II) in the corpora
cardiaca and the hemolymph of Leptinotarsa decemlineata enormously increases in
the adults fed on genetically modified potatoes containing either GNA lectin or
Cry 3Aa toxin concomitant with increased oxidative stress in gut tissues. A
similar enhancement of the AKH titer is achieved when the adults are injected
with paraquat that evokes oxidative stress. On the other hand, an injection of
exogenous AKH reduces oxidative stress biomarkers in the hemolymph by reducing
protein carbonyls and enhancing reduced glutathione levels. These facts indicate
that there is a feedback regulation between an oxidative stressor action and the
level of AKH in the insect body, and that AKHs might be involved in the
activation of an antioxidant protection mechanism. These results are to our
knowledge, the first evidence for the involvement of AKHs in oxidative stress
mitigation, in addition to a plethora of other roles.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17353065 [PubMed - indexed for MEDLINE]

724: Plant Physiol. 2007 May;144(1):18-31. Epub 2007 Mar 9.

Successful gene tagging in lettuce using the Tnt1 retrotransposon from tobacco.

Mazier M, Botton E, Flamain F, Bouchet JP, Courtial B, Chupeau MC, Chupeau Y,
Maisonneuve B, Lucas H.

Unitö© de Gö©nö©tique et d'Amö©lioration des Fruits et Lö©gumes, UR1502, Institut
National de la Recherche Agronomique, F-84143 Montfavet cedex, France.
mazier@avignon.inra.fr

The tobacco (Nicotiana tabacum) element Tnt1 is one of the few identified active
retrotransposons in plants. These elements possess unique properties that make
them ideal genetic tools for gene tagging. Here, we demonstrate the feasibility
of gene tagging using the retrotransposon Tnt1 in lettuce (Lactuca sativa), which
is the largest genome tested for retrotransposon mutagenesis so far. Of 10
different transgenic bushes carrying a complete Tnt1 containing T-DNA, eight
contained multiple transposed copies of Tnt1. The number of transposed copies of
the element per plant was particularly high, the smallest number being 28. Tnt1
transposition in lettuce can be induced by a very simple in vitro culture
protocol. Tnt1 insertions were stable in the progeny of the primary transformants
and could be segregated genetically. Characterization of the sequences flanking
some insertion sites revealed that Tnt1 often inserted into genes. The progeny of
some primary transformants showed phenotypic alterations due to recessive
mutations. One of these mutations was due to Tnt1 insertion in the gibberellin
3beta-hydroxylase gene. Taken together, these results indicate that Tnt1 is a
powerful tool for insertion mutagenesis especially in plants with a large genome.

PMID: 17351058 [PubMed - indexed for MEDLINE]

725: Plant Physiol. 2007 May;144(1):380-90. Epub 2007 Mar 9.

A WUSCHEL-LIKE HOMEOBOX gene represses a YABBY gene expression required for rice
leaf development.

Dai M, Hu Y, Zhao Y, Liu H, Zhou DX.

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural
University, Wuhan 430070, China.

YABBY and WUSCHEL-LIKE HOMEOBOX (WOX) genes have been shown to play important
roles in lateral organ formation and meristem function. Here, we report the
characterization of functional relationship between rice (Oryza sativa) YAB3 and
WOX3 in rice leaf development. Rice YAB3 is closely related to maize (Zea mays)
ZmYAB14 and Arabidopsis (Arabidopsis thaliana) FILAMENTOUS FLOWER (FIL), whereas
rice WOX3 is highly conserved with maize narrow sheath1 (NS1) and NS2 and
Arabidopsis PRESSED FLOWER (PRS). In situ hybridization experiments revealed that
the expression of both genes was excluded from the shoot apical meristem, but the
transcripts were detected in leaf primordia, young leaves, and reproductive
organs without any polar distribution. The function of the two genes was studied
by both overexpression and RNA interference (RNAi) in transgenic rice. YAB3 RNAi
induced twisted and knotted leaves lacking specialized structures such as ligule
and auricles, while no phenotypic change was observed in YAB3 overexpression
plants, suggesting that rice YAB3 may be required for leaf cell growth and
differentiation. Overexpression of WOX3 repressed YAB3 and showed a YAB3 RNAi
phenotype. The expression of class I KNOTTED-LIKE HOMEOBOX (KNOX) genes was
ectopically induced in leaves of YAB3 RNAi or WOX3 overexpression plants. Data
from inducible WOX3 expression and DNA-protein interaction assays suggested that
WOX3 acted as a transcriptional repressor of YAB3. These data reveal a regulatory
network involving YAB3, WOX3, and KNOX genes required for rice leaf development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17351053 [PubMed - indexed for MEDLINE]

726: Environ Entomol. 2007 Feb;36(1):234-44.

Transgenes sustain epigeal insect biodiversity in diversified vegetable farm
systems.

Leslie TW, Hoheisel GA, Biddinger DJ, Rohr JR, Fleischer SJ.

Department of Entomology, Pennsylvania State University, PA 16802, USA.
twl117@psu.edu

Many ecological studies have focused on the effects of transgenes in field crops,
but few have considered multiple transgenes in diversified vegetable systems. We
compared the epigeal, or soil surface-dwelling, communities of Coleoptera and
Formicidae between transgenic and isoline vegetable systems consisting of sweet
corn, potato, and acorn squash, with transgenic cultivars expressing Cry1(A)b,
Cry3, or viral coat proteins. Vegetables were grown in replicated split plots
over 2 yr with integrated pest management (IPM) standards defining insecticide
use patterns. More than 77.6% of 11,925 insects from 1,512 pitfall traps were
identified to species, and activity density was used to compare dominance
distribution, species richness, and community composition. Measures of epigeal
biodiversity were always equal in transgenic vegetables, which required fewer
insecticide applications than their near isolines. There were no differences in
species richness between transgenic and isoline treatments at the farm system and
individual crop level. Dominance distributions were also similar between
transgenic and isoline farming systems. Crop type, and not genotype, had a
significant influence on Carabidae and Staphylinidae community composition in the
first year, but there were no treatment effects in the second year, possibly
because of homogenizing effects of crop rotations. Communities were more
influenced by crop type, and possibly crop rotation, than by genotype. The
heterogeneity of crops and rotations in diversified vegetable farms seems to aid
in preserving epigeal biodiversity, which may be supplemented by reductions in
insecticide use associated with transgenic cultivars.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17349138 [PubMed - indexed for MEDLINE]

727: Environ Entomol. 2007 Feb;36(1):228-33.

Effects of Cry1Ab-expressing corn anthers on the movement of monarch butterfly
larvae.

Prasifka PL, Hellmich RL, Prasifka JR, Lewis LC.

Department of Entomology, 13 Insectary Bldg., Iowa State University, Ames, IA
50011, USA. anderpl@iastate.edu

Decreased larval feeding and weight of the monarch butterfly, Danaus plexippus
L., have been detected after 4 d of exposure in the laboratory to a high density
of Bacillus thuringiensis (Bt)-expressing anthers. One hypothesis is that larvae
exposed to Bt anthers exhibit increased wandering, resulting in less feeding and
lower weight gain. To test this hypothesis, 2-d-old monarch butterfly larvae
exposed to milkweed leaf disks with no anthers, anthers that express Bt (Cry1Ab,
event MON810), or other non-Bt anthers were observed using a video-tracking
system. As had been shown in previous studies, larvae exposed to Bt anthers fed
less and gained less weight than larvae exposed to non-Bt or no anthers, yet
there was no evidence of feeding on anthers. Total distance moved, maximum
displacement from release point, percentage of time spent moving or near anthers,
or mean turn angle did not differ across treatments. However, larvae exposed to
Bt anthers spent more time off milkweed leaf disks than those exposed to no
anthers and were more likely to move off the leaf than larvae exposed to non-Bt
anthers. Results suggest that larvae exposed to Bt anthers behave differently and
that ingestion may not be the only way Bt can affect nontarget insects like the
monarch butterfly.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17349137 [PubMed - indexed for MEDLINE]

728: J Agric Food Chem. 2007 Apr 4;55(7):2582-9. Epub 2007 Mar 10.

Diverted secondary metabolism and improved resistance to European corn borer
(Ostrinia nubilalis) in maize (Zea mays L.) transformed with wheat oxalate
oxidase.

Mao J, Burt AJ, Ramputh AI, Simmonds J, Cass L, Hubbard K, Miller S, Altosaar I,
Arnason JT.

Department of Biology, University of Ottawa, 30 Marie Curie Street, Ottawa,
Ontario, Canada K1N 6N5.

An alteration in the secondary metabolism of maize (Zea mays L.) genetically
modified with the wheat oxalate oxidase (OxO) gene was observed using HPLC and
fluorescence microscopy. Phenolic concentrations in the OxO lines were
significantly increased, but DIMBOA synthesis was reduced due to a diversion in
the shikimate pathway leading to phenolic and hydroxamic acids. Ferulic acid
exhibited the largest increase and accounted for 80.4% of the total soluble
phenolics. Transcription of a 13-lipoxygenase gene, coding for a key enzyme
involved in the regulation of secondary metabolism, was substantially higher in
the OxO line than in the null line. To test whether the high levels of soluble
phenolic acids, in particular ferulic acid, contributed to the insect resistance
in the OxO maize, ferulic acid was administered in meridic diets to European corn
borer (ECB). A significant negative correlation between ferulic acid
concentration and ECB larval growth rate was found. Field testing during 2001
showed that OxO maize was more resistant to ECB, with leaf consumption and
stalk-tunneling damage significantly reduced by 28-34 and 37-39%, respectively,
on all of the OxO lines tested and confirming published 2000 findings.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17348672 [PubMed - indexed for MEDLINE]

729: Sci Am. 2007 Mar;296(3):8.

The beef with cloned meat.

[No authors listed]

PMID: 17348147 [PubMed - indexed for MEDLINE]

730: Science. 2007 Mar 23;315(5819):1712-6. Epub 2007 Mar 8.

Comment in:
Science. 2007 Mar 23;315(5819):1676-7. Science. 2007 Nov 9;318(5852):914; author reply 914.

A G protein-coupled receptor is a plasma membrane receptor for the plant hormone
abscisic acid.

Liu X, Yue Y, Li B, Nie Y, Li W, Wu WH, Ma L.

National Institute of Biological Sciences, 7 Science Park Road, Zhongguancun Life
Science Park, Beijing 102206, China.

The plant hormone abscisic acid (ABA) regulates many physiological and
developmental processes in plants. The mechanism of ABA perception at the cell
surface is not understood. Here, we report that a G protein-coupled receptor
genetically and physically interacts with the G protein alpha subunit GPA1 to
mediate all known ABA responses in Arabidopsis. Overexpressing this receptor
results in an ABA-hypersensitive phenotype. This receptor binds ABA with high
affinity at physiological concentration with expected kinetics and
stereospecificity. The binding of ABA to the receptor leads to the dissociation
of the receptor-GPA1 complex in yeast. Our results demonstrate that this G
protein-coupled receptor is a plasma membrane ABA receptor.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17347412 [PubMed - indexed for MEDLINE]

731: Int J Immunopathol Pharmacol. 2007 Jan-Mar;20(1):111-8.

Longer resistance of some DNA traits from BT176 maize to gastric juice from
gastrointestinal affected patients.

Ferrini AM, Mannoni V, Pontieri E, Pourshaban M.

Istituto Superiore di Sanitö , National Centre for Food Quality and Risk
Assessment, Rome, Italy. ferrini@iss.it

The presence of antibiotic resistance marker genes in genetically engineered
plants is one of the most controversial issues related to Genetically Modified
Organism (GMO)-containing food, raising concern about the possibility that these
markers could increase the pool of antibiotic resistance genes. This study
investigates the in vitro survival of genes bla and cryIA(b) of maize Bt176 in
human gastric juice samples. Five samples of gastric juice were collected from
patients affected by gastro-esophageal reflux or celiac disease and three
additional samples were obtained by pH modification with NaHCO3. DNA was
extracted from maize Bt176 and incubated with samples of gastric juices at
different times. The survival of the target traits (bla gene, whole 1914 bp gene
cry1A(b), and its 211 bp fragment) was determined using PCR. The stability of the
target genes was an inverse function of their lengths in all the samples.
Survival in samples from untreated subjects was below the normal physiological
time of gastric digestion. On the contrary, survival time in samples from
patients under anti-acid drug treatment or in samples whose pH was modified,
resulted strongly increased. Our data indicate the possibility that in particular
cases the survival time could be so delayed that, as a consequence, some traits
of DNA could reach the intestine. In general, this aspect must be considered for
vulnerable consumers (people suffering from gastrointestinal diseases related to
altered digestive functionality, physiological problems or drug side-effects) in
the risk analysis usually referred to healthy subjects.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17346434 [PubMed - indexed for MEDLINE]

732: Plant J. 2007 Apr;50(1):14-28. Epub 2007 Mar 5.

An NB-LRR protein required for HR signalling mediated by both extra- and
intracellular resistance proteins.

Gabriö«ls SH, Vossen JH, Ekengren SK, van Ooijen G, Abd-El-Haliem AM, van den Berg
GC, Rainey DY, Martin GB, Takken FL, de Wit PJ, Joosten MH.

Laboratory of Phytopathology, Wageningen University, Binnenhaven 5, 6709 PD
Wageningen, The Netherlands.

Tomato (Solanum lycopersicum) Cf resistance genes confer hypersensitive response
(HR)-associated resistance to strains of the pathogenic fungus Cladosporium
fulvum that express the matching avirulence (Avr) gene. Previously, we identified
an Avr4-responsive tomato (ART) gene that is required for Cf-4/Avr4-induced HR in
Nicotiana benthamiana as demonstrated by virus-induced gene silencing (VIGS). The
gene encodes a CC-NB-LRR type resistance (R) protein analogue that we have
designated NRC1 (NB-LRR protein required for HR-associated cell death 1). Here we
describe that knock-down of NRC1 in tomato not only affects the Cf-4/Avr4-induced
HR but also compromises Cf-4-mediated resistance to C. fulvum. In addition, VIGS
using NRC1 in N. benthamiana revealed that this protein is also required for the
HR induced by the R proteins Cf-9, LeEix, Pto, Rx and Mi. Transient expression of
NRC1(D481V), which encodes a constitutively active NRC1 mutant protein, triggers
an elicitor-independent HR. Subsequently, we transiently expressed this
auto-activating protein in N. benthamiana silenced for genes known to be involved
in HR signalling, thereby allowing NRC1 to be positioned in an HR signalling
pathway. We found that NRC1 requires RAR1 and SGT1 to be functional, whereas it
does not require NDR1 and EDS1. As the Cf-4 protein requires EDS1 for its
function, we hypothesize that NRC1 functions downstream of EDS1. We also found
that NRC1 acts upstream of a MAP kinase pathway. We conclude that Cf-mediated
resistance signalling requires a downstream NB-LRR protein that also functions in
cell death signalling pathways triggered by other R proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17346268 [PubMed - indexed for MEDLINE]

733: Plant J. 2007 Apr;50(1):54-69. Epub 2007 Mar 5.

Regulation and functional analysis of ZmDREB2A in response to drought and heat
stresses in Zea mays L.

Qin F, Kakimoto M, Sakuma Y, Maruyama K, Osakabe Y, Tran LS, Shinozaki K,
Yamaguchi-Shinozaki K.

Biological Resources Division, Japan International Research Center for
Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 305-8686, Japan.

DREB1/CBFs and DREB2s are transcription factors that specifically interact with a
cis-acting element, DRE/CRT, which is involved in the expression of genes
responsive to cold and drought stress in Arabidopsis thaliana. The function of
DREB1/CBFs has been precisely analyzed and it has been found to activate the
expression of many genes responsive to cold stress containing a DRE/CRT sequence
in their promoters. However, the regulation and function of DREB2-type
transcription factors remained to be elucidated. In this research, we report the
cloning of a DREB2 homolog from maize, ZmDREB2A, whose transcripts were
accumulated by cold, dehydration, salt and heat stresses in maize seedlings.
Unlike Arabidopsis DREB2A, ZmDREB2A produced two forms of transcripts, and
quantitative real-time PCR analyses demonstrated that only the functional
transcription form of ZmDREB2A was significantly induced by stresses. Moreover,
the ZmDREB2A protein exhibited considerably high transactivation activity
compared with DREB2A in Arabidopsis protoplasts, suggesting that protein
modification is not necessary for ZmDREB2A to be active. Constitutive or
stress-inducible expression of ZmDREB2A resulted in an improved drought stress
tolerance in plants. Microarray analyses of transgenic plants overexpressing
ZmDREB2A revealed that in addition to genes encoding late embryogenesis abundant
(LEA) proteins, some genes related to heat shock and detoxification were also
upregulated. Furthermore, overexpression of ZmDREB2A also enhanced
thermotolerance in transgenic plants, implying that ZmDREB2A may play a dual
functional role in mediating the expression of genes responsive to both water
stress and heat stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17346263 [PubMed - indexed for MEDLINE]

734: Nat Biotechnol. 2007 Mar;25(3):282-3.

Comment on:
Nat Biotechnol. 2007 Jan;25(1):1.

Two views of the emperor's new clones.

Schubert D.

Publication Types:
Comment
Letter

PMID: 17344873 [PubMed - indexed for MEDLINE]

735: Nat Biotechnol. 2007 Mar;25(3):281.

Comment on:
Nat Biotechnol. 2007 Jan;25(1):1.

Two views of the emperor's new clones.

Miller HI.

Publication Types:
Comment
Letter

PMID: 17344872 [PubMed - indexed for MEDLINE]

736: Nat Biotechnol. 2007 Mar;25(3):271.

Agbiotech booms in emerging nations.

Lawrence S.

PMID: 17344870 [PubMed - indexed for MEDLINE]

737: Heredity. 2007 Jun;98(6):375-84. Epub 2007 Mar 7.

Recent degeneration of an old duplicated flowering time gene in Brassica nigra.

Sjö¶din P, Hedman H, Shavorskaya O, Finet C, Lascoux M, Lagercrantz U.

Evolutionary Functional Genomics, Evolutionary Biology Centre, Uppsala
University, Uppsala, Sweden.

Gene and genome duplications play a major role in the evolution of plant species.
The Brassica nigra genome is highly replicated as a result of ancient
polyploidization events. Two copies of the flowering time gene CONSTANS (COa and
COb) have been identified in B. nigra, and previous studies showed that COa is
functional. In the present study, the polymorphism of 92 COb alleles sampled in
seven populations was analyzed. Both polymorphism and recombination levels were
elevated and varied strongly among populations and 8% of COb alleles exhibit
apparently disabling mutations. Sequence data, however, do not provide
unambiguous support for the presence of relaxed selective constraint on COb as
compared to known functional CO genes. On the one hand, some of the disabling
mutations reached high-frequency arguing for a loss of function but, on the other
hand, the ratio of nonsynonymous to synonymous nucleotide polymorphism and
diversity is low and similar to that observed in other B. nigra CO and CO-like
genes, supporting the conservation of some function. We also showed that COb is
still transcribed. Finally, the flowering time of Arabidopsis thaliana co mutant
plants transformed with COb alleles with and without apparent disabling mutations
was similar. We propose that COb was retained for a long period after
duplication, but a recent fixation of a detrimental mutation, possibly as an
effect of a bottleneck, resulted in its nonfunctionalization. We also speculate
as to the presence of subsequent selection for rapid degeneration of the gene.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17344804 [PubMed - indexed for MEDLINE]

738: Am J Physiol Renal Physiol. 2007 Jun;292(6):F1858-66. Epub 2007 Mar 6.

Renal vascular and tubulointerstitial inflammation and proliferation in
Cyp1a1-Ren2 transgenic rats with inducible ANG II-dependent malignant
hypertension.

Graciano ML, Mouton CR, Patterson ME, Seth DM, Mullins JJ, Mitchell KD.

Department of Physiology, Hypertension and Renal Center of Excellence, Tulane
University Health Sciences Center, New Orleans, Louisiana 70112, USA.

Transgenic rats with inducible ANG II-dependent malignant hypertension
[TGR(Cyp1a1Ren2)] were generated by inserting the mouse Ren2 renin gene into the
genome of the rat. The present study was performed to assess renal morphological
changes occurring during the development of ANG II-dependent malignant
hypertension in these rats. Male Cyp1a1-Ren2 rats (n = 10) were fed normal rat
food containing indole-3-carbinol (I3C; 0.3%) for 10 days to induce malignant
hypertension. Rats induced with I3C had higher mean arterial pressures (173 +/- 9
vs. 112 +/- 11 mmHg, P < 0.01) than noninduced normotensive rats (n = 9).
Glomerular damage was evaluated by determination of the glomerulosclerosis index
(GSI) in tissue sections stained with periodic acid-Schiff. Kidneys of
hypertensive rats had a higher GSI than normotensive rats (21.3 +/- 5.6 vs. 3.5
+/- 1.31 units). Quantitative analysis of macrophage ED-1-positive cells and
proliferating cell nuclear antigen using immunohistochemistry demonstrated
increased macrophage numbers in the renal interstitium (106.4 +/- 11.4 vs. 58.7
+/- 5.0 cells/mm(2)) and increased proliferating cell number in cortical tubules
(37.8 +/- 5.7 vs. 24.2 +/- 2.1 cells/mm(2)), renal cortical vessels (2.2 +/- 0.5
vs. 0.13 +/- 0.07 cells/vessel), and the cortical interstitium (33.6 +/- 5.7 vs.
4.2 +/- 1.4 cells/mm(2)) of hypertensive rat kidneys. These findings demonstrate
that the renal pathological changes that occur during the development of
malignant hypertension in Cyp1a1-Ren2 rats are characterized by inflammation and
cellular proliferation in cortical vessels and tubulointerstitium.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17344186 [PubMed - indexed for MEDLINE]

739: Pest Manag Sci. 2007 May;63(5):440-6.

Pyramiding of insecticidal compounds for control of the cowpea bruchid
(Callosobruchus maculatus F.).

Tarver MR, Shade RE, Shukle RH, Moar WJ, Muir WM, Murdock LM, Pittendrigh BR.

Department of Entomology, Purdue University, West Lafayette, IN 47901-1158, USA.

The cowpea bruchid (Callosobruchus maculatus F.) (Chrysomelidae: Bruchini) is a
major pest of stored cowpea grain. With limited available technologies for
controlling the bruchid, transgenic cowpeas with bruchid resistance genes
engineered into them could become the next management tools. An investigation was
made of two different sets of potential transgenic insecticidal compounds using
an artificial seed system: (i) CIP-PH-BT-J and recombinant egg white avidin, and
(ii) avidin and wheat alpha-amylase inhibitor. CIP-PH-BT-J (0.1%; 1000 mg kg(-1))
and recombinant egg white avidin (0.006%; 60 mg kg(-1)) incorporated separately
into artificial seeds caused 98.2 and 99% larval mortality rates respectively.
Combining CIP-PH-BT-J and avidin in the same artificial seed provided additional
mortality compared with each factor incorporated singly; no insects survived in
seeds with the combined toxins. Similarly, when avidin and wheat alpha-amylase
inhibitor (alphaAI) (1%; 10 g kg(-1)) were incorporated separately into
artificial seeds, this caused 99.8 and 98% mortality respectively. However, in
combination, avidin and alphaAI did not increase mortality, but they did cause a
significant increase in developmental time of the cowpea bruchids. These results
emphasize that the joint action of potential insecticidal compounds cannot be
predicted from results obtained separately for each compound, and they suggest
potential transgenes for further consideration.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17340671 [PubMed - indexed for MEDLINE]

740: Toxicol Sci. 2007 May;97(1):27-31. Epub 2007 Mar 3.

The utility of an international sera bank for use in evaluating the potential
human allergenicity of novel proteins.

Thomas K, Bannon G, Herouet-Guicheney C, Ladics G, Lee L, Lee SI, Privalle L,
Ballmer-Weber B, Vieths S.

International Life Sciences Institute Health and Environmental Sciences
Institute, Washington, District of Columbia 20005, USA. kthomas@ilsi.org

In the safety assessment of novel foods produced through biotechnology, careful
consideration is given to determining the allergenic potential of newly
introduced proteins. IgE serum screening is one tool for evaluating whether the
protein in question has sequence identity to a known allergen or if the source of
the gene encoding the protein is a known allergenic food. A "specific" serum
screen involves testing a gene product with sera from patients with documented
clinical allergy to a specific allergen to confirm that the gene product of
interest is not the same protein to which the patient produces IgE antibodies. A
"targeted" serum screen involves testing the gene product of interest with sera
from patients sensitive to food or aeroallergens from the same broad group. The
concept of a global sera bank with accessible, well-characterized sera for use in
such assays is an appealing option. This paper summarizes the consensus elements
from a workshop to evaluate the potential utility of an international sera bank
for evaluating the allergenicity of novel proteins. Areas of agreement following
the workshop included the following: (1) specific sera screens are appropriate
for exploring potentially cross-reactive proteins that have been identified
through bioinformatics analyses; however, additional validation is needed,
particularly for targeted sera screens, (2) practical and ethical considerations
may preclude the formation of a global sera bank, and therefore, (3) a regional
network of clinicians who could serve as sources of patient sera or be approached
to conduct sera studies would be the most practical alternative.

Publication Types:
Congresses
Research Support, Non-U.S. Gov't

PMID: 17337755 [PubMed - indexed for MEDLINE]

741: J Exp Biol. 2007 Mar;210(Pt 6):956-63.

The role of larval fat cells in adult Drosophila melanogaster.

Aguila JR, Suszko J, Gibbs AG, Hoshizaki DK.

School of Life Sciences, University of Nevada Las Vegas, 4505 Maryland Parkway,
Las Vegas, NV 89154, USA.

In the life history of holometabolous insects, distinct developmental stages are
tightly linked to feeding and non-feeding periods. The larval stage is
characterized by extensive feeding, which supports the rapid growth of the animal
and allows accumulation of energy stores, primarily in the larval fat body. In
Drosophila melanogaster access to these stores during pupal development is
possible because the larval fat body is preserved in the pupa as individual fat
cells. These larval fat cells are refractive to autophagic cell death that
removes most of the larval cells during metamorphosis. The larval fat cells are
thought to persist into the adult stage and thus might also have a nutritional
role in the young adult. We used cell markers to demonstrate that the fat cells
in the young adult are in fact dissociated larval fat body cells, and we present
evidence that these cells are eventually removed in the adult by a caspase
cascade that leads to cell death. By genetically manipulating the lifespan of the
larval fat cells, we demonstrate that these cells are nutritionally important
during the early, non-feeding stage of adulthood. We experimentally blocked cell
death of larval fat cells using the GAL4/UAS system and found that in newly
eclosed adults starvation resistance increased from 58 h to 72 h. Starvation
survival was highly correlated with the number of remaining larval fat cells. We
discuss the implications of these results in terms of the overall nutritional
status of the larva as an important factor in adult survival in environmental
stresses such as starvation.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17337708 [PubMed - indexed for MEDLINE]

742: Protein Expr Purif. 2007 Jun;53(2):325-30. Epub 2007 Jan 23.

Large scale production of Bacillus thuringiensis PS149B1 insecticidal proteins
Cry34Ab1 and Cry35Ab1 from Pseudomonas fluorescens.

Huang KX, Badger M, Haney K, Evans SL.

Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN 46268, USA.
khhuang@dow.com

The 14kDa (Cry34Ab1) and 44kDa (Cry35Ab1) binary insecticidal proteins are
produced naturally by Bacillus thuringiensis PS149B1 as parasporal inclusion
bodies. Here, we show production of these two insecticidal proteins in
recombinant Pseudomonas fluorescens and their subsequent purification to near
homogeneity to provide large quantities of protein for safety-assessment studies
associated with the registration of transgenic corn plants. The gene sequence
specific for each protein was expressed in P. fluorescens and fermented at the
75-L scale. For Cry34Ab1, the protein accumulated as insoluble inclusion bodies,
and was purified by extraction directly from the cell pastes at pH 3.4 with a
sodium acetate buffer, selective precipitation at pH 7.0, and differential
centrifugation. For Cry35Ab1, the protein was extracted from the purified
inclusion bodies with sodium acetate buffer (pH 3.5) containing 0.5M urea,
followed by diafiltration. No chromatography steps were required to produce over
30g of lyophilized protein powder with purity greater than 98%, while retaining
full insecticidal activity against Western corn rootworm larvae. The proteins
were further characterized to assure identity and suitability for use in
safety-assessment studies.

PMID: 17337206 [PubMed - indexed for MEDLINE]

743: New Phytol. 2007;174(1):63-76.

In vivo visualization of F-actin structures during the development of the moss
Physcomitrella patens.

Finka A, Schaefer DG, Saidi Y, Goloubinoff P, Zrö¿d JP.

SV/IBI, EPFL, Station 15, CH-1015 Lausanne, Switzerland.

* The 'in planta' visualization of F-actin in all cells and in all developmental
stages of a plant is a challenging problem. By using the soybean heat inducible
Gmhsp17.3B promoter instead of a constitutive promoter, we have been able to
label all cells in various developmental stages of the moss Physcomitrella
patens, through a precise temperature tuning of the expression of green
fluorescent protein (GFP)-talin. * A short moderate heat treatment was sufficient
to induce proper labeling of the actin cytoskeleton and to allow the
visualization of time-dependent organization of F-actin structures without
impairment of cell viability. * In growing moss cells, dense converging arrays of
F-actin structures were present at the growing tips of protonema cell, and at the
localization of branching. Protonema and leaf cells contained a network of thick
actin cables; during de-differentiation of leaf cells into new protonema
filaments, the thick bundled actin network disappeared, and a new highly
polarized F-actin network formed. * The controlled expression of GFP-talin
through an inducible promoter improves significantly the 'in planta' imaging of
actin.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17335498 [PubMed - indexed for MEDLINE]

744: Arch Virol. 2007;152(7):1323-39. Epub 2007 Mar 5.

Making a friend from a foe: expressing a GroEL gene from the whitefly Bemisia
tabaci in the phloem of tomato plants confers resistance to tomato yellow leaf
curl virus.

Akad F, Eybishtz A, Edelbaum D, Gorovits R, Dar-Issa O, Iraki N, Czosnek H.

The Otto Warburg Minerva Center for Agricultural Biotechnology & The Robert H
Smith Institute for Plant Science and Genetics in Agriculture, The Hebrew
University of Jerusalem, Rehovot, Israel.

Some (perhaps all) plant viruses transmitted in a circulative manner by their
insect vectors avoid destruction in the haemolymph by interacting with GroEL
homologues, ensuring transmission. We have previously shown that the
phloem-limited begomovirus tomato yellow leaf curl virus (TYLCV) interacts in
vivo and in vitro with GroEL produced by the whitefly vector Bemisia tabaci. In
this study, we have exploited this phenomenon to generate transgenic tomato
plants expressing the whitefly GroEL in their phloem. We postulated that
following inoculation, TYLCV particles will be trapped by GroEL in the plant
phloem, thereby inhibiting virus replication and movement, thereby rendering the
plants resistant. A whitefly GroEL gene was cloned in an Agrobacterium vector
under the control of an Arabidopsis phloem-specific promoter, which was used to
transform two tomato genotypes. During three consecutive generations, plants
expressing GroEL exhibited mild or no disease symptoms upon whitefly-mediated
inoculation of TYLCV. In vitro assays indicated that the sap of resistant plants
contained GroEL-TYLCV complexes. Infected resistant plants served as virus source
for whitefly-mediated transmission as effectively as infected non-transgenic
tomato. Non-transgenic susceptible tomato plants grafted on resistant
GroEL-transgenic scions remained susceptible, although GroEL translocated into
the grafted plant and GroEL-TYLCV complexes were detected in the grafted tissues.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17334947 [PubMed - indexed for MEDLINE]

745: Exp Appl Acarol. 2007;41(3):191-201. Epub 2007 Mar 3.

Toxicological evaluation of genetically modified cotton (Bollgard) and Dipel WP
on the non-target soil mite Scheloribates praeincisus (Acari: Oribatida).

Oliveira AR, Castro TR, Capalbo DM, Delalibera I Jr.

Department of Entomology, Plant Pathology and Agricultural Zoology, University of
Sao Paulo, CP 9, 13418-900 Piracicaba, SP, Brazil.

Insecticides derived from the bacterium Bacillus thuringiensis (Bt) and plants
genetically modified (GM) to express B. thuringiensis toxins are important
alternatives for insect pest control worldwide. Risk assessment of B.
thuringiensis toxins to non-target organisms has been extensively studied but few
toxicological tests have considered soil invertebrates. Oribatid mites are one of
the most diverse and abundant arthropod groups in the upper layers of soil and
litter in natural and agricultural systems. These mites are exposed to the toxic
compounds of GM crops or pesticides mainly when they feed on vegetal products
incorporated in the soil. Although some effects of B. thuringiensis products on
Acari have been reported, effects on oribatid mites are still unknown. This study
investigated the effects of the ingestion of Bt cotton Bollgard and of the B.
thuringiensis commercial product Dipel WP on the pantropical species
Scheloribates praeincisus (Scheloribatidae). Ingestion of Bollgard and Dipel did
not affect adult and immature survivorship and food consumption (estimated by
number of fecal pellets produced daily) or developmental time of immature stages
of S. praeincisus. These results indicate the safety of Bollgard and Dipel to S.
praeincisus under field conditions where exposition is lower and other food
sources besides leaves of Bt plants are available. The method for toxicological
tests described here can be adapted to other species of Oribatida, consisting on
a new option to risk assessment studies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17334814 [PubMed - indexed for MEDLINE]

746: Plant Mol Biol. 2007 May;64(1-2):35-47. Epub 2007 Mar 2.

The expression of iron homeostasis-related genes during rice germination.

Nozoye T, Inoue H, Takahashi M, Ishimaru Y, Nakanishi H, Mori S, Nishizawa NK.

Graduate School of Agricultural and Life Sciences, The University of Tokyo,
Tokyo, Japan.

To characterize Fe homeostasis during the early stages of seed germination, a
microarray analysis was performed. mRNAs extracted from fully mature seeds or
seeds harvested 1-3 days after sowing were hybridized to a rice microarray
containing approximately 22,000 cDNA oligo probes. Many Fe deficiency-inducible
genes were strongly expressed throughout early seed germination. These results
suggest that the demand for Fe is extremely high during germination.Under
Fe-deficient conditions, rice produces and secretes a metal-cation chelator
called deoxymugineic acid (DMA) to acquire Fe from the soil. In addition, DMA and
its intermediate nicotianamine (NA) are thought to be involved in long distance
Fe transport in rice. Using promoter-beta-glucuronidase (GUS) analysis, we
investigated the expression patterns during seed germination of the Fe
deficiency-inducible genes OsNAS1, OsNAS2, OsNAS3, OsNAAT1, and OsDMAS1, which
encode enzymes that participate in the biosynthesis of DMA, and the transporter
genes OsYSL2 and OsIRT1, which are involved in Fe transport. All of these genes
were expressed in germinating seeds prior to protrusion of the radicle. These
results suggest that DMA and NA are produced and involved in Fe transport during
germination.

PMID: 17333504 [PubMed - indexed for MEDLINE]

747: J Chem Ecol. 2007 Apr;33(4):669-81. Epub 2007 Feb 28.

Systemin regulates both systemic and volatile signaling in tomato plants.

Corrado G, Sasso R, Pasquariello M, Iodice L, Carretta A, Cascone P, Ariati L,
Digilio MC, Guerrieri E, Rao R.

Dipartimento di Scienze del Suolo della Pianta e dell'Ambiente, Universitö  degli
Studi di Napoli Federico II, 80055, Portici, Naples, Italy.

The prevailing reaction of plants to pest attack is the activation of various
defense mechanisms. In tomato, several studies indicate that an 18 amino acid
(aa) peptide, called systemin, is a primary signal for the systemic induction of
direct resistance against plant-chewing pests, and that the transgenic expression
of the prosystemin gene (encoding the 200 aa systemin precursor) activates genes
involved in the plant response to herbivores. By using a combination of
behavioral, chemical, and gene expression analyses, we report that systemin
enhances the production of bioactive volatile compounds, increases plant
attractivity towards parasitiod wasps, and activates genes involved in volatile
production. Our data imply that systemin is involved in the systemic activation
of indirect defense in tomato, and we conclude that a single gene controls the
systemic activation of coordinated and associated responses against pests.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17333376 [PubMed - indexed for MEDLINE]

748: Plant Cell Rep. 2007 Jul;26(7):1035-44. Epub 2007 Feb 27.

Non-antibiotic, efficient selection for alfalfa genetic engineering.

Rosellini D, Capomaccio S, Ferradini N, Savo Sardaro ML, Nicolia A, Veronesi F.

Dipartimento di Biologia Vegetale Biotecnologie Agroambientali e Zootecniche,
Universitö  degli Studi di Perugia, Borgo XX giugno 74, 06121 Perugia, Italy.
roselli@unipg.it

A selectable marker gene (SMG), usually conferring resistance to an antibiotic or
herbicide, is generally introduced into the plant cells with the gene(s) for the
trait of interest to allow only the cells that have integrated and express the
foreign sequences to regenerate into a plant. The availability of several SMGs
for each plant species is useful for both basic and applied research to combine
several genes of interest in the same plant. A selection system based on
gabaculine (3-amino-2,3-dihydrobenzoic acid) as the selective substance and the
bacterial hemL gene [encoding a mutant for of the enzyme glutamate 1-semialdehyde
aminotransferase (GSA-AT)] as the SMG was previously used for genetic
transformation of tobacco. The hemL gene is a good candidate for a safe SMG,
because GSA-AT is present in all plants and is likely involved in one metabolic
step only, so that unintended effects of its overexpression in plants are not
probable. In this work, we have compared this new selection system with the
conventional, kanamycin-based system for alfalfa Agrobacterium-mediated
transformation. The hemL and NptII genes were placed together into a T-DNA under
the control of identical promoters and terminators. We show that the
gabaculine-based system is more efficient than the conventional, kanamycin-based
system. The inheritance of hemL was Mendelian, and no obvious phenotypic effect
of its expression was observed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17333020 [PubMed - indexed for MEDLINE]

749: Plant Cell Rep. 2007 Jul;26(7):1011-23. Epub 2007 Mar 1.

Chloroplast targeting of FanC, the major antigenic subunit of Escherichia coli
K99 fimbriae, in transgenic soybean.

Garg R, Tolbert M, Oakes JL, Clemente TE, Bost KL, Piller KJ.

Department of Biology, University of North Carolina-Charlotte, 9201 University
City Boulevard, Charlotte, NC 28223, USA.

Enterotoxigenic Escherichia coli (ETEC) strains are a major cause of enteric
diseases affecting livestock and humans. Edible transgenic plants producing E.
coli fimbrial subunit proteins have the potential to vaccinate against these
diseases, but have not reached their full potential as a renewable source of oral
vaccines due in part to insufficient levels of recombinant protein accumulation.
Previously, we reported that cytosol targeting of the E. coli K99 fimbrial
subunit antigen resulted in FanC accumulation to approximately 0.4% of total
soluble protein in soybean leaves (Piller et al. in Planta 222:6-18, 2005). In
this study, we report on the subcellular targeting of FanC to chloroplasts.
Twenty-two transgenic T1 progeny derived from seven individual T0 transformation
events were characterized, and 17 accumulated transgenic FanC. All of the
characterized events displayed relatively low T-DNA complexity, and all exhibited
proper targeting of FanC to the chloroplast. Accumulation of chloroplast-targeted
FanC was approximately 0.08% of total soluble leaf protein, or approximately
5-fold less than cytosol-targeted FanC. Protein analysis of leaves at various
stages of maturity suggested stability of chloroplast-targeted FanC throughout
leaf maturation. Furthermore, mice immunized intraperitoneally with protein
extract derived from transgenic leaves expressing chloroplast-targeted FanC
developed significant antibody titers against FanC. This is the first report of
subcellular targeting of a vaccine subunit antigen in soybean.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 17333019 [PubMed - indexed for MEDLINE]

750: Plant Cell. 2007 Feb;19(2):433-44. Epub 2007 Feb 28.

Comment in:
Plant Cell. 2007 Feb;19(2):391-3.

The absence of histone H2B monoubiquitination in the Arabidopsis hub1 (rdo4)
mutant reveals a role for chromatin remodeling in seed dormancy.

Liu Y, Koornneef M, Soppe WJ.

Department of Plant Breeding and Genetics, Max Planck Institute for Plant
Breeding Research, 50829 Cologne, Germany.

Seed dormancy is defined as the failure of a viable seed to germinate under
favorable conditions. Besides playing an adaptive role in nature by optimizing
germination to the most suitable time, a tight control of dormancy is important
in crop plants. Extensive genetic and physiological studies have identified the
involvement of several factors, but the molecular mechanisms underlying this
process are still largely unknown. We cloned the HISTONE MONOUBIQUITINATION1
(HUB1) gene, of which the mutant (previously identified as reduced dormancy4) has
reduced seed dormancy and several pleiotropic phenotypes. HUB1 encodes a C3HC4
RING finger protein. The Arabidopsis thaliana genome contains one HUB1 homolog,
which we named HUB2. The hub2 mutant also has reduced seed dormancy and is not
redundant with hub1. Homologs of HUB1 and HUB2 in other species are required for
histone H2B monoubiquitination. In agreement with this, the ubiquitinated form of
histone H2B could not be detected in the hub1 and hub2 mutants. In yeast and
human cells, histone H2B monoubiquitination is associated with actively
transcribed genes. The hub1 mutant showed altered expression levels for several
dormancy-related genes. We propose a role for chromatin remodeling in seed
dormancy by H2B monoubiquitination through HUB1 and HUB2.

PMID: 17329563 [PubMed - indexed for MEDLINE]

751: Food Chem Toxicol. 2007 Jul;45(7):1277-92. Epub 2007 Jan 25.

Subchronic feeding study of DAS-59122-7 maize grain in Sprague-Dawley rats.

Malley LA, Everds NE, Reynolds J, Mann PC, Lamb I, Rood T, Schmidt J, Layton RJ,
Prochaska LM, Hinds M, Locke M, Chui CF, Claussen F, Mattsson JL, Delaney B.

DuPont Haskell Laboratory, Newark, DE, United States.

59122 is a transgenic maize line containing event DAS-59122-7 that expresses the
corn rootworm (CRW) specific pesticidal Cry34Ab1 and Cry35Ab1 proteins from
Bacillus thuringiensis (Bt) Berliner strain PS149B1 and the
phosphinothricin-N-acetyltransferase (PAT) protein from Streptomyces
viridochromogenes for tolerance to the herbicidal ingredient
glufosinate-ammonium. For the current study, 59122 maize grain, non-transgenic
near-isogenic maize grain (091), and a commercially available non-transgenic
reference maize grain (33R77) were grown under conditions simulating commercial
farming practices. Adult Sprague-Dawley rats (12/sex/group) were fed diets
formulated with 35% maize grain from either 59122, 091, or 33R77, or one of two
separate lots of commercially available rodent chow prepared with commercially
available corn (35%) in accordance with the standards of Purina Mills Labdiet
5002 for approximately 90 days. All diets possessed similar nutritional and
contaminant profiles. The transgenic proteins were detected only in diets
prepared with 59122 maize grain and were stable over the course of the study.
Compared to control groups, no adverse diet-related differences were observed in
rats fed diets formulated with 59122 maize grain with respect to body
weight/gain, food consumption/efficiency, clinical signs of toxicity, mortality,
ophthalmology, neurobehavioral (FOB and motor activity) assessments, clinical
pathology (hematology, clinical chemistry, coagulation, and urinalysis), and
pathology (organ weights and gross and microscopic pathology). Results from this
study indicate that 59122 maize grain is nutritionally equivalent to and as safe
as conventional maize grain.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17329002 [PubMed - indexed for MEDLINE]

752: Environ Biosafety Res. 2006 Apr-Jun;5(2):77-87. Epub 2006 Dec 8.

Detection of feral transgenic oilseed rape with multiple-herbicide resistance in
Japan.

Aono M, Wakiyama S, Nagatsu M, Nakajima N, Tamaoki M, Kubo A, Saji H.

Environmental Biology Division, National Institute for Environmental Studies,
16-2 Onogawa, Tsukuba, 305-8506, Japan. maono@nies.go.jp

Repeated monitoring for escaped transgenic crop plants is sometimes necessary,
especially in cases when the crop has not been approved for release into the
environment. Transgenic oilseed rape (Brassica napus) was detected along
roadsides in central Japan in a previous study. The goal of the current study was
to monitor the distribution of transgenic oilseed rape and occurrence of
hybridization of transgenic B. napus with feral populations of its closely
related species (B. rapa and B. juncea) in the west of Japan in 2005. The
progenies of 50 B. napus, 82 B. rapa and 283 B. juncea maternal plants from 95
sampling sites in seven port areas were screened for herbicide-resistance.
Transgenic herbicide-resistant seeds were detected from 12 B. napus maternal
plants growing at seven sampling sites in two port areas. A portion of the
progeny from two transgenic B. napus plants had both glyphosate-resistance and
glufosinate-resistance transgenes. Therefore, two types of transgenic B. napus
plants are likely to have outcrossed with each other, since the
double-herbicide-resistant transgenic strain of oilseed rape has not been
developed intentionally for commercial purposes. As found in the previous study,
no transgenic seeds were detected from B. rapa or B. juncea, and more extensive
sampling is needed to determine whether introgression into these wild species has
occurred.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17328854 [PubMed - indexed for MEDLINE]

753: J Proteome Res. 2007 Apr;6(4):1354-63. Epub 2007 Feb 28.

Seeking transformation markers: an analysis of differential tissue proteomes on
the rice germplasm generated from transformation of Echinochloa crusgalli genomic
DNA.

Zhao C, Zhao B, Ren Y, Tong W, Wang J, Zhao K, Shu S, Xu N, Liu S.

Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300, China.

The transformation of distally related genomic DNAs into plant was proposed as a
novel technique to breed new cultivars. For example, a restorer rice line, RB207,
was successfully developed and stabilized through the transformation of genomic
DNAs of Echinochloa crusgalli (E. crusgalli) into a rice line, R207. Although the
phenotypes of this variant line are apparently different from its receptor, the
molecular bases are not elucidated yet. Herein, we have systematically studied
the differential proteomes from the tissues of E. crusgalli, R207, and RB207 in
an attempt to find an explanation regarding the phenotypic changes of RB207. The
2-DE method was employed to separate the leaf and embryo proteins of these plants
followed by protein identification with mass spectrometry. In the leaf, 953 +/-
15, 1084 +/- 11, and 1091 +/- 11 silver-stained spots were detected, whereas in
the embryo, 986 +/- 3, 884 +/- 10, and 892 +/- 14 spots were found from E.
crusgalli, R207, and RB207, respectively. In comparison to the 2-DE images of the
two rice lines, which showed many similarities, the ones of the E. crusgalli and
rice were found to be so different that they were incomparable. There were some
differentially expressed 2-DE spots between the two rice cultivars, 72 in leaf
and 53 in embryo, respectively. The results of protein identification suggested
that, regardless of leaves or embryos, none of the E. crusgalli genes were
encoded in the new rice cultivar, RB207. The fact that 60% of the differentially
expressed spots between R207 and RB207, however, were verified as the proteins
involved in metabolism and photosynthesis makes a rather convincing argument that
the DNA fragments transferred from E. crusgalli to rice are responsible for
exerting the unknown influence to the expression of rice genes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17326673 [PubMed - indexed for MEDLINE]

754: J Agric Food Chem. 2007 Apr 4;55(7):2509-16. Epub 2007 Feb 28.

Identification of PCR-amplified genetically modified organisms (GMOs) DNA by
peptide nucleic acid (PNA) probes in anion-exchange chromatographic analysis.

Rossi S, Lesignoli F, Germini A, Faccini A, Sforza S, Corradini R, Marchelli R.

Dipartimento di Chimica Organica e Industriale Universitö  di Parma, Viale G. P.
Usberti 17/A, I-43100 Parma, Italy.

PCR products obtained by selective amplification of transgenic DNA derived from
food samples containing Roundup Ready soybean or Bt-176 maize have been analyzed
by anion-exchange HPLC. Peptide nucleic acids (PNAs), oligonucleotide analogues
known to bind to complementary single-stranded DNA with high affinity and
specificity, have been used as specific probes in order to assess the identity of
the peaks observed. Two different protocols were adopted in order to obtain
single-stranded DNA: amplification with an excess of one primer or digestion of
one DNA strand. The single-stranded DNA was mixed with the PNA probe, and the
presence of a specific sequence was revealed through detection of the
corresponding PNA:DNA peak with significantly different retention time.
Advantages and limits of this approach are discussed. The method was tested with
reference materials and subsequently applied to commercial samples.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17326652 [PubMed - indexed for MEDLINE]

755: Plant Cell Environ. 2007 Apr;30(4):508-17.

Effect of hexokinase activity on tomato root metabolism during prolonged hypoxia.

Gharbi I, Ricard B, Rolin D, Maucourt M, Andrieu MH, Bizid E, Smiti S, Brouquisse
R.

Laboratoire de Physiologie Vö©gö©tale, Dö©partement de Biologie, Facultö© des
Sciences de Tunis, 1060 Tunisia.

Hypoxically induced tolerance to anoxia in roots of tomato (Solanum lycopersicum)
was previously shown to depend on sucrose and the induction of sucrose synthase.
In contrast to maize, root hexokinase (HXK) activities did not increase during
hypoxia and glucose was unable to sustain glycolytic flux under anoxia. In this
paper, we asked whether hypoxic metabolism in roots would be altered in
transgenic tomato plants overexpressing either a plant (Arabidopsis) or a yeast
(Saccharomyces cerevisiae) HXK and whether such modifications could be related to
improved energy metabolism and consequently root tolerance under anoxia. Tomato
plants grown hydroponically with shoots always maintained in air were submitted
to a 7 d hypoxic treatment applied by stopping air bubbling. A combination of
techniques including (1)H-nuclear magnetic resonance spectroscopy, RT-PCR and
enzyme analyses was used to obtain a broad picture of hypoxic root metabolism. In
normoxic conditions, HXK overexpression resulted in higher ADP and AMP levels
only in roots of AtHXK1 transgenic plants. During hypoxic treatment, oxygen
levels in the hydroponic tank decreased rapidly to 5 kPa within the first 2 d and
then remained at 5 kPa throughout the 7 d experiment. Oxygen levels were similar
at 5 and 20 cm below the water surface. A decline of the adenylate energy status
was observed after 2 d of hypoxic treatment, with a further decrease by 7 d in
roots of non-transgenic (WT) and ScHXK2, but not in AtHXK1 transgenic plants.
Sucrose synthase activity increased to comparably higher levels at 7 d of hypoxic
treatment in WT and ScHXK2 compared with AtHXK1 roots. Differences between WT and
the transgenic plants are discussed with respect to the metabolic response to low
(hypoxia) but not zero (anoxia) oxygen.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17324236 [PubMed - indexed for MEDLINE]

756: J Agric Food Chem. 2007 Mar 21;55(6):2452-8. Epub 2007 Feb 27.

The influence of diluted seawater and ripening stage on the content of
antioxidants in fruits of different tomato genotypes.

Sgherri C, Navari-Izzo F, Pardossi A, Soressi GP, Izzo R.

Dipartimento di Chimica e Biotecnologie Agrarie, Universitö  di Pisa, Via del
Borghetto 80, Italy.

The aim of this study was to investigate if the combined effect of diluted
seawater and ripening can improve the beneficial nutritional properties of tomato
fruits from an antioxidant point of view. To reach the goal, different tomato
cultivars and breeding lines, genetically modified for ripening, were
investigated, and analysis of NADPH and NADP+ as well as of the main antioxidants
such as ascorbic acid, lipoic acid, and tocopherols was performed at two ripening
stages. The research was conducted on berries of the following genotypes of
tomato: cv. Jama, Gimar wild type, Gimar gf, and Gimar nor. The mutant gf is a
typical "stay green" mutant, characterized by an incomplete loss of chlorophyll;
the nor mutation is characterized by a reduced biosynthesis of ethylene and
carotenoids. Both ripening and salinity induced an oxidative stress, and the
sensitivity to salt treatment was genotype-dependent. The genotypes cv. Jama and
Gimar gf line showed increases in ascorbic acid, lipoic acid, and
alpha-tocopherol during both ripening and salt treatment whereas total ascorbate
and tocopherols decreased in the berries from salt-treated plants of Gimar wild
type. Ripening also determined decreases in ascorbate and tocopherol amounts in
the Gimar nor line where a positive effect of ripening and salinity was observed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17323974 [PubMed - indexed for MEDLINE]

757: Planta. 2007 Jul;226(2):429-42. Epub 2007 Feb 24.

Characterization of vascular-specific RSs1 and rolC promoters for their
utilization in engineering plants to develop resistance against hemipteran insect
pests.

Saha P, Chakraborti D, Sarkar A, Dutta I, Basu D, Das S.

Plant Molecular and Cellular Genetics, Bose Institute, P1/12 C.I.T. Scheme VIIM,
Kolkata 700054, West Bengal, India.

Rice sucrose synthase1, RSs1 (isolated from rice) and rolC (isolated from
Agrobacterium rhizogenes) promoters were evaluated by binding analyses of their
respective cis-elements with host nuclear transcription factors. The expression
profile of an insecticidal protein driven by these promoters in transgenic plants
was monitored. Motif-search analysis with available phloem-specific promoter
sequences revealed the presence of two BoxII elements in RSs1. An octopine
synthase element, a stem-specific, a root-specific and a light-responsive element
were found in the rolC promoter, whereas the ASL box, GATA and 13 bp motifs were
detected in both promoters. Binding analysis of these cis-elements (both in
native and mutant forms) with the trans-factors present in the nuclear extracts
from rice, tobacco and chickpea, followed by electrophoretic mobility shift
assay, documented a highly specific cis-trans interaction. Both promoters were
utilized to express Allium sativum leaf agglutinin (ASAL) gene in the three
aforementioned plant systems. By immunohistochemistry and
immunohistofluorescence, specific patterns of ASAL accumulation were detected in
vascular tissues of single copy transgenic plants. Transgenic plants expressing
ASAL in a phloem-specific manner demonstrated about 60-65% more insecticidal
activity than control plants. The two promoters, which evolved independently from
two distinctly unrelated origins, were found to maintain their functionality in a
conserved manner. They were able to express the insecticidal protein coding ASAL
as transgene both in monocot and dicot hosts. Thus, the two promoters are
valuable as prospective phloem-specific promoters for use in plant
biotechnological programmes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17323077 [PubMed - indexed for MEDLINE]

758: Science. 2007 Feb 23;315(5815):1069.

Environmental regulation. U.S. courts say transgenic crops need tighter scrutiny.

Charles D.

Publication Types:
News

PMID: 17322039 [PubMed - indexed for MEDLINE]

759: Food Chem Toxicol. 2007 Jul;45(7):1179-85. Epub 2007 Jan 11.

Safety assessment of transgenic Bacillus thuringiensis with VIP insecticidal
protein gene by feeding studies.

Peng D, Chen S, Ruan L, Li L, Yu Z, Sun M.

State Key Laboratory of Agricultural Microbiology, College of Life Science and
Technology, Huazhong Agricultural University, Wuhan 430070, People's Republic of
China.

The aim of this study was to evaluate the toxicology safety of a genetically
modified (GM) Bacillus thuringiensis with vegetative insecticidal protein (VIP)
gene. Acute and subacute toxicity studies by using its powder preparation were
conducted in Wistar rats. The result of the acute study showed the
no-observable-adverse-effect level (NOAEL) of this GM B. thuringiensis powder
preparation was greater than 5000 mg/kg body weight (BW). In the subacute study,
the data analysis of body weight gain, food and water consumptions, clinical
observations, haematology, serum biochemistry, organ weight ratios and
histopathological findings did not show significant differences between control
and treated groups. These results proved the NOAEL of this GM B. thuringiensis
powder preparation in subacute test was greater than 5000 mg/kg BW. Since both
the acute and subacute oral toxicity were not detected at the highest dose
recommended by OECD guidelines, this GM B. thuringiensis could be generally
regarded as safe for use in bio-pesticide industry.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17320261 [PubMed - indexed for MEDLINE]

760: Mol Genet Genomics. 2007 Jun;277(6):713-23. Epub 2007 Feb 21.

Over-expression of calcium-dependent protein kinase 13 and calreticulin
interacting protein 1 confers cold tolerance on rice plants.

Komatsu S, Yang G, Khan M, Onodera H, Toki S, Yamaguchi M.

National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba 305-8518, Japan.
skomatsu@affrc.go.jp

Calcium is a ubiquitous signaling molecule and changes in cytosolic calcium
concentration are involved in plant responses to various stimuli. The rice
calcium-dependent protein kinase 13 (CDPK13) and calreticulin interacting protein
1 (CRTintP1) have previously been reported to be involved in cold stress response
in rice. In this study, rice lines transformed with sense CDPK13 or CRTintP1
constructs were produced and used to investigate the function of these proteins.
When the plants were incubated at 5 degrees C for 3 days, leaf blades of both the
sense transgenic and vector control rice plants became wilted and curled. When
the plants were transferred back to non-stress conditions after cold treatment,
the leaf blades died, but the sheaths remained green in the sense transgenic rice
plants. Expression of CDPK13 or CRTintP1 was further examined in several rice
varieties including cold-tolerant rice varieties. Accumulation of these proteins
in the cold-tolerant rice variety was higher than that in rice varieties that are
intermediate in their cold tolerance. To examine whether over-expression of
CDPK13 and CRTintP1 would have any effect on the proteins or not, sense
transgenic rice plants were analyzed using proteomics. The 2D-PAGE profiles of
proteins from the vector control were compared with those of the sense transgenic
rice plants. Two of the proteins that differed between these lines were
calreticulins. The results suggest that CDPK13, calreticulin and CRTintP1 might
be important signaling components for response to cold stress in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17318583 [PubMed - indexed for MEDLINE]

761: Plant J. 2007 Mar;49(5):935-46.

Hormonal control of the inflated calyx syndrome, a morphological novelty, in
Physalis.

He C, Saedler H.

Department of Molecular Plant Genetics, Max-Planck-Institute for Plant Breeding
Research, Carl-von-Linnö©-Weg 10, D-50829 Cologne, Germany.

The 'Chinese lantern' phenotype or inflated calyx syndrome (ICS)--inflated sepals
encapsulating the mature berry of Physalis floridana--is a morphological novelty
within the Solanaceae. ICS is associated with heterotopic expression of MPF2,
which codes for a MADS-box transcription factor otherwise involved in leaf
formation and male fertility. In accordance with this finding, the MPF2 promoter
sequence differs significantly from that of its orthologue STMADS16 in the
related Solanum tuberosum, which does not exhibit ICS. However, heterotopic
expression of MPF2 is not sufficient for ICS formation in P. floridana-
fertilization is also important. Here we report that the hormones cytokinin and
gibberellin are essential for ICS formation. MPF2 controls sepal cell division,
but the resulting cells are small. Calyx size increases substantially only if
gibberellin and cytokinin are available to promote cell elongation and further
cell division. Transient expression of appropriate MPF2-/STMADS16-GFP fusions in
leaf tissues in the presence of hormones revealed that cytokinin, but not
gibberellin, facilitated transport of the transcription factor into the nucleus.
Furthermore, an ICS-like structure can be induced in transgenic S. tuberosum by
ectopic expression of STMADS16 and simultaneous treatment with cytokinin and
gibberellin. Strikingly, transgenic Arabidopsis ectopically expressing
solanaceous MPF2-like proteins display enhanced sepal growth when exposed to
cytokinin only, while orthologous proteins from non-solanaceous plants did not
require cytokinin for this function. These data are incorporated into a detailed
model for ICS formation in P. floridana.

PMID: 17316177 [PubMed - indexed for MEDLINE]

762: Clin Vaccine Immunol. 2007 Apr;14(4):464-9. Epub 2007 Feb 21.

Expression of the human hepatitis B virus large surface antigen gene in
transgenic tomato plants.

Lou XM, Yao QH, Zhang Z, Peng RH, Xiong AS, Wang HK.

Shanghai Key Laboratory of Agricultural Genetics and Breeding, Agro-Biotechnology
Research Center, Shanghai Academy of Agricultural Sciences, 2901 Beidi Rd.,
Shanghai 201106, P. R. China.

The original hepatitis B virus (HBV) large surface antigen gene was synthesized.
In order to optimize the expression of this gene in tomato plants, the tobacco
pathogenesis-related protein S signal peptide was fused to the 5' end of the
modified gene and the sequence encoding amino acids S, E, K, D, E, and L was
placed at the 3' end. The gene encoding the modified HBV large surface antigen
under the control of a fruit-specific promoter was constructed and expressed in
transgenic tomato plants. The expression of the antigen from transgenic plants
was confirmed by PCR and reverse transcriptase PCR. Enzyme-linked immunoassays
using a monoclonal antibody directed against human serum-derived HBsAg revealed
that the maximal level of HBsAg was about 0.02% of the soluble protein in
transgenic tomato fruit. The amount of HBsAg in mature fruits was found to be 65-
to 171-fold larger than in small or medium fruits and leaf tissues. Examination
of transgenic plant samples by transmission electron microscopy proved that HBsAg
had been expressed and had accumulated. The HBsAg protein was capable of
assembling into capsomers and virus-like particles. To our knowledge, this is the
first time the HBV large surface antigen has been expressed in plants. This work
suggests the possibility of producing a new alternative vaccine for human HBV.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17314228 [PubMed - indexed for MEDLINE]

763: Protein Expr Purif. 2007 Jun;53(2):270-4. Epub 2007 Jan 18.

Reduced protease activity in transformed rice cell suspension cultures expressing
a proteinase inhibitor.

Kim TG, Kim HM, Lee HJ, Shin YJ, Kwon TH, Lee NJ, Jang YS, Yang MS.

Division of Biological Sciences and the Research Center for Bioactive Materials,
Chonbuk National University, Jeonju 561-756, Republic of Korea.

In this study, we synthesized a synthetic serine proteinase inhibitor II gene
(sPI-II) that harbored the chymotrypsin and trypsin inhibitor domains of the
PI-II gene from Nicotiana alata. In an effort to reduce protease activity in a
rice cell suspension culture, we first synthesized sPI-II using overlap PCR and
then introduced the gene into a rice calli (Oryza sativa L. cv. Dongin) by
particle bombardment-mediated transformation. The sPI-II gene was under the
control of a rice alpha-amylase 3D promoter induced by sugar starvation. To
verify the integration and expression of the sPI-II gene in the transformed rice
cells, we employed genomic DNA PCR amplification and Northern blot analysis,
respectively. The relative protease activity of the transformed cell suspension
culture was reduced to approximately 23% when compared to the non-transformed
culture. This indicates that a transformed suspension culture system expressing a
proteinase inhibitor, may be a useful tool to protect against recombinant protein
losses resulting from extracellular proteases.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17314052 [PubMed - indexed for MEDLINE]

764: Mol Plant Microbe Interact. 2007 Feb;20(2):107-19.

GmEREBP1 is a transcription factor activating defense genes in soybean and
Arabidopsis.

Mazarei M, Elling AA, Maier TR, Puthoff DP, Baum TJ.

Department of Plant Pathology, Iowa State University, Ames 50011, USA.

Ethylene-responsive element-binding proteins (EREBPs) are plant-specific
transcription factors, many of which have been linked to plant defense responses.
Conserved EREBP domains bind to the GCC box, a promoter element found in
pathogenesis-related (PR) genes. We previously identified an EREBP gene from
soybean (GmEREBP1) whose transcript abundance decreased in soybean
cyst-nematode-infected roots of a susceptible cultivar, whereas it increased in
abundance in infected roots of a resistant cultivar. Here, we report further
characterization of this gene. Transient expression analyses showed that GmEREBP1
is localized to the plant nucleus and functions as a transcriptional activator in
soybean leaves. Transgenic soybean plants expressing GmEREBP1 activated the
expression of the ethylene (ET)-responsive gene PR2 and the ET- and jasmonic acid
(JA)-responsive gene PR3, and the salicylic acid (SA)-responsive gene PR1 but not
the SA-responsive PR5. Similarly, transgenic Arabidopsis plants expressing
GmEREBP1 showed elevated mRNA abundance of the ET-regulated gene PR3 and the ET-
and JA-regulated defense-related gene PDF1.2 but not the ET-regulated GST2, and
the SA-regulated gene PR1 but not the SA-regulated PR2 and PR5. Transgenic
soybean and Arabidopsis plants inoculated with cyst nematodes did not display a
significantly altered susceptibility to nematode infection. These results
collectively show that GmEREBP1 functions as a transacting inducer of defense
gene expression in both soybean and Arabidopsis and mediates the expression of
both ET- and JA- and SA-regulated defense-related genes in these plant species.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17313162 [PubMed - indexed for MEDLINE]

765: J Agric Food Chem. 2007 Mar 21;55(6):2115-20. Epub 2007 Feb 21.

Analysis of bioavailable arsenic in rice with whole cell living bioreporter
bacteria.

Baumann B, van der Meer JR.

Department of Fundamental Microbiology, University of Lausanne, CH 1015 Lausanne,
Switzerland.

A multiwell plate bioassay was developed using genetically modified bacteria
(bioreporter cells) to detect inorganic arsenic extracted from rice. The
bacterial cells expressed luciferase upon exposure to arsenite, the activity of
which was detected by measurement of cellular bioluminescence. The bioreporter
cells detected arsenic in all rice varieties tested, with averages of 0.02-0.15
microg of arsenite equivalent per gram of dry weight and a method detection limit
of 6 ng of arsenite per gram of dry rice. This amounted to between approximately
20 and 90% of the total As content reported by chemical methods for the same
sample and suggested that a major proportion of arsenic in rice is in the
inorganic form. Calibrations of the bioassay with pure inorganic and organic
arsenic forms showed that the bacterial cells react to arsenite with highest
affinity, followed by arsenate (with 25% response relative to an equivalent
arsenite concentration) and trimethylarsine oxide (at 10% relative response). A
method for biocompatible arsenic extraction was elaborated, which most optimally
consisted of (i) grinding rice to powder, (ii) mixing with an aqueous solution
containing pancreatic enzymes, (iii) mechanical shearing, (iv) extraction in mild
acid conditions and moderate heat, and (v) centrifugation and pH neutralization.
Detection of mainly inorganic arsenic by the bacterial cells may have important
advantages for toxicity assessment of rice consumption and would form a good
complement to total chemical arsenic determination.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17311403 [PubMed - indexed for MEDLINE]

766: Lab Anim (NY). 2007 Mar;36(3):8.

Hens as protein bioreactors.

Schub T.

Publication Types:
News

PMID: 17311035 [PubMed - indexed for MEDLINE]

767: Environ Sci Technol. 2007 Jan 15;41(2):599-605.

Transgenic Indian mustard overexpressing selenocysteine lyase or selenocysteine
methyltransferase exhibit enhanced potential for selenium phytoremediation under
field conditions.

Baö±uelos G, LeDuc DL, Pilon-Smits EA, Terry N.

ARS-USDA, 9611 South Riverbend Avenue, Parlier, California 93648, USA.

Two new transgenic Indian mustard [Brassica juncea (L.) Czern.] lines were tested
under field conditions for their ability to accumulate selenium (Se)from Se- and
boron-contaminated saline sediment. The transgenic lines overexpress genes
encoding the enzymes selenocysteine lyase (cpSL) and selenocysteine
methyltransferase (SMT), respectively. In the first Spring planting, cpSL, SMT,
and wildtype plants (WT) were compared, while SMT and WT were compared in a
second, Fall planting. In the Spring planting, shoots of the cpSL transgenic
plants accumulated 2-fold more Se (p < 0.01), had 1.8 times higher leaf Se
concentrations (p < 0.01), and grew better on contaminated soil than WT. The SMT
plants had a 1.7-fold higher leaf Se concentration than WT (p < 0.05). In the
Fall planting, the SMT transgenic plants accumulated 1.6-fold more Se in their
shoots than WT (p < 0.01) with Se concentrations being higher in both leaves and
stems. These results conclusively demonstrate that cpSL and SMT transgenic lines
have significantly greater Se phytoremediation potential than wildtype Indian
mustard. Further, this study confirms the importance of field testing for
evaluating future transgenic lines.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17310728 [PubMed - indexed for MEDLINE]

768: Plant Cell Rep. 2007 Jul;26(7):969-76. Epub 2007 Feb 20.

Expression of Escherichia coli heat-labile enterotoxin b subunit (LTB) in carrot
(Daucus carota L.).

Rosales-Mendoza S, Soria-Guerra RE, de Jesöºs Olivera-Flores MT, Lö¨pez-Revilla R,
Argö¼ello-Astorga GR, Jimö©nez-Bremont JF, Garcö­a-de la Cruz RF, Loyola-Rodrö­guez
JP, Alpuche-Solö­s AG.

Divisiö¨n de Biologö­a Molecular, Instituto Potosino de Investigaciö¨n Cientö­fica y
Tecnolö¨gica, Camino a la Presa San Josö© 2055, 78216 San Luis Potosö­, S.L.P.,
Mexico.

We expressed the B subunit of enterotoxigenic Escherichia coli heat-labile
enterotoxin (LTB) encoded by a synthetic codon-optimized gene in carrot. An
Agrobacterium-mediated transformation method was used. Thirty independent
transgenic lines were regenerated via somatic embryogenesis after 6 months in
culture and were transferred to a greenhouse. GM1-ELISA assay was used to assess
LTB protein content in mature taproots. Some transgenic lines expressed LTB up to
0.3% of the total soluble protein, which is tenfold higher than the expression
levels reported earlier using the native bacterial gene in plants. Immunological
assay confirmed proper assembly of the pentameric complex and in vitro activity
of the recombinant LTB protein, suggesting that it can be functional in
prevention of diarrhea.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17310334 [PubMed - indexed for MEDLINE]

769: Plant Biotechnol J. 2006 Nov;4(6):667-76.

A large-scale field study of transgene flow from cultivated rice (Oryza sativa)
to common wild rice (O. rufipogon) and barnyard grass (Echinochloa crusgalli).

Wang F, Yuan QH, Shi L, Qian Q, Liu WG, Kuang BG, Zeng DL, Liao YL, Cao B, Jia
SR.

Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou
51064, China.

The introgression of transgenes into wild relatives or weeds through
pollen-mediated gene flow is a major concern in environmental risk assessment of
transgenic crops. A large-scale (1.3-1.8 ha) rice gene flow study was conducted
using transgenic rice containing the bar gene as a pollen donor and Oryza
rufipogon as a recipient. There was a high frequency of transgene flow (11%-18%)
at 0-1 m, with a steep decline with increasing distance to a detection limit of
0.01% by 250 m. To our knowledge, this is the highest frequency and longest
distance of gene flow from transgenic rice to O. rufipogon reported so far. On
the basis of these data, an adequate isolation distance from both conventional
and transgenic rice should be taken for in situ conservation of common wild rice.
Meanwhile, there is no evidence of transgene introgression into barnyard grass,
even when it has coexisted with transgenic rice containing the bar gene for five
successive years. Thus, the environmental risk of gene flow to this weedy species
is of little concern.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17309736 [PubMed - indexed for MEDLINE]

770: Plant Biotechnol J. 2006 Nov;4(6):633-45.

Pollen-mediated gene flow in maize in real situations of coexistence.

Messeguer J, Peö±as G, Ballester J, Bas M, Serra J, Salvia J, Palaudelmö s M, Melö©
E.

Consorci Laboratori CSIC-IRTA de Genö¨tica Molecular Vegetal, Departament de
Genö¨tica Vegetal, Centre de Cabrils, Carretera de Cabrils s/n, Cabrils 08348
(Barcelona), Spain. joaquima.messeguer@irta.es

We present the first study on cross-fertilization between Bt and conventional
maize in real situations of coexistence in two regions in which Bt and
conventional maize were cultivated. A map was designed and the different crops
were identified, as were the sowing and flowering dates, in Bt and conventional
maize fields. These data were used to choose the non-transgenic fields for
sampling and analysis by the real-time quantification system-polymerase chain
reaction (RTQ-PCR) technique. In general, the rate of cross-fertilization was
higher in the borders and, in most of the fields, decreased towards the centre of
the field. Nine fields had values of genetically modified organism DNA to total
DNA of much lower than 0.9%, whereas in three the rate was higher. Some
differences were found when comparing our results with those of common field
trials. In real conditions of coexistence and in cropping areas with smaller
fields, the main factors that determined cross-pollination were the synchronicity
of flowering and the distances between the donor and receptor fields. By
establishing an index based on these two variables, the rate of the adventitious
presence of genetically modified maize could be predicted, as well as the
influence of other factors. By applying this index, and in the case of a fully
synchronous flowering time, a security distance between transgenic and
conventional fields of about 20 m should be sufficient to maintain the
adventitious presence of genetically modified organisms as a result of pollen
flow below the 0.9% threshold in the total yield of the field.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17309734 [PubMed - indexed for MEDLINE]

771: Plant Biotechnol J. 2006 Sep;4(5):529-49.

Constitutive expression of the pea ABA-responsive 17 (ABR17) cDNA confers
multiple stress tolerance in Arabidopsis thaliana.

Srivastava S, Rahman MH, Shah S, Kav NN.

Department of Agricultural, Food and Nutritional Science, University of Alberta,
Edmonton, AB, Canada, T6G 2P5.

The constitutive expression of the pea ABR17 (abscisic acid-responsive 17) cDNA,
which is a member of the group 10 family of pathogenesis-related proteins (PR
10), in Arabidopsis thaliana is reported. The presence of ABR17 transcripts and
the protein in the three transgenic lines is demonstrated by reverse
transcriptase-polymerase chain reaction (RT-PCR) and two-dimensional
electrophoresis followed by tandem mass spectrometry, respectively. Three
independently derived transgenic lines containing ABR17 germinated better in the
presence of salt, cold temperature or both. Furthermore, the transgenic plants
also exhibited enhanced tolerance to freezing temperature, suggesting the
potential utility of the ABR17 gene to engineer multiple stress tolerance. In
order to obtain insights into the mechanism underlying ABR17-mediated stress
tolerance, we have compared the proteome of a transgenic line with that of its
wild-type counterpart. Several proteins were observed to be significantly altered
in the transgenic line, including some with a role(s) in photosynthesis, stress
tolerance and the regulation of gene expression. Our findings are discussed
within the context of available genes to engineer multiple stress tolerance as
well as the biological activities of the ABR17 protein.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17309728 [PubMed - indexed for MEDLINE]

772: Plant Biotechnol J. 2006 Sep;4(5):499-510.

High accumulation of bioactive peptide in transgenic rice seeds by expression of
introduced multiple genes.

Wakasa Y, Yasuda H, Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, Kannondai 3-1-3, Tsukuba, Ibaraki 305-8604, Japan.

We have developed a simple binary vector construction system for the simultaneous
expression of multiple genes in plants. Up to three independent gene cassettes
can be easily integrated into one binary vector using the MultiSite Gateway
System. Using this system, we produced transgenic rice plants that accumulated
high levels of the hypocholesterolaemic peptide lactostatin (IIAEK) in endosperm.
Binary vectors were constructed that could accommodate up to three independent
modified glutelin gene cassettes encoding multimer lactostatin in the variable
regions. Eight construct permutations were used for rice transformation. We
measured the accumulation of lactostatin expressed as a glutelin fusion protein
in the mature seeds of 105 independent transgenic rice lines. A general
correlation was observed between accumulation level and gene number in the vector
constructs, indicating that a higher accumulation of lactostatin was obtained
from transgenic rice plants containing the maximum number of gene inserts. These
results indicate that this strategy is applicable for the selection of transgenic
lines containing large amounts of bioactive peptides in rice seeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17309726 [PubMed - indexed for MEDLINE]

773: Plant Biotechnol J. 2007 Mar;5(2):313-24.

Functional analysis of rice NPR1-like genes reveals that OsNPR1/NH1 is the rice
orthologue conferring disease resistance with enhanced herbivore susceptibility.

Yuan Y, Zhong S, Li Q, Zhu Z, Lou Y, Wang L, Wang J, Wang M, Li Q, Yang D, He Z.

National Key Laboratory of Plant Molecular Genetics, Institute of Plant
Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences, Shanghai 200032, China.

The key regulator of salicylic acid (SA)-mediated resistance, NPR1, is
functionally conserved in diverse plant species, including rice (Oryza sativa
L.). Investigation in depth is needed to provide an understanding of
NPR1-mediated resistance and a practical strategy for the improvement of disease
resistance in the model crop rice. The rice genome contains five NPR1-like genes.
In our study, three rice homologous genes, OsNPR1/NH1, OsNPR2/NH2 and OsNPR3,
were found to be induced by rice bacterial blight Xanthomonas oryzae pv. oryzae
and rice blast Magnaporthe grisea, and the defence molecules benzothiadiazole,
methyl jasmonate and ethylene. We confirmed that OsNPR1 is the rice orthologue by
complementing the Arabidopsis npr1 mutant. Over-expression of OsNPR1 conferred
disease resistance to bacterial blight, but also enhanced herbivore
susceptibility in transgenic plants. The OsNPR1-green fluorescent protein (GFP)
fusion protein was localized in the cytoplasm and moved into the nucleus after
redox change. Mutations in its conserved cysteine residues led to the
constitutive localization of OsNPR1(2CA)-GFP in the nucleus and also abolished
herbivore hypersensitivity in transgenic rice. Different subcellular
localizations of OsNPR1 antagonistically regulated SA- and jasmonic acid
(JA)-responsive genes, but not SA and JA levels, indicating that OsNPR1 might
mediate antagonistic cross-talk between the SA- and JA-dependent pathways in
rice. This study demonstrates that rice has evolved an SA-mediated systemic
acquired resistance similar to that in Arabidopsis, and also provides a practical
approach for the improvement of disease resistance without the penalty of
decreased herbivore resistance in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17309686 [PubMed - indexed for MEDLINE]

774: Plant Biotechnol J. 2007 Mar;5(2):263-274.

'GM-gene-deletor': fused loxP-FRT recognition sequences dramatically improve the
efficiency of FLP or CRE recombinase on transgene excision from pollen and seed
of tobacco plants.

Luo K, Duan H, Zhao D, Zheng X, Deng W, Chen Y, Stewart CN Jr, McAvoy R, Jiang X,
Wu Y, He A, Pei Y, Li Y.

Department of Plant Science, University of Connecticut, Storrs, CT 06269, USA.

Pollen- and seed-mediated transgene flow is a concern in plant biotechnology. We
report here a highly efficient 'genetically modified (GM)-gene-deletor' system to
remove all functional transgenes from pollen, seed or both. With the three
pollen- and/or seed-specific gene promoters tested, the phage CRE/loxP or yeast
FLP/FRT system alone was inefficient in excising transgenes from tobacco pollen
and/or seed, with no transgenic event having 100% efficiency. When loxP-FRT
fusion sequences were used as recognition sites, simultaneous expression of both
FLP and CRE reduced the average excision efficiency, but the expression of FLP or
CRE alone increased the average excision efficiency, with many transgenic events
being 100% efficient based on more than 25,000 T(1) progeny examined per event.
The 'GM-gene-deletor' reported here may be used to produce 'non-transgenic'
pollen and/or seed from transgenic plants and to provide a bioconfinement tool
for transgenic crops and perennials, with special applicability towards
vegetatively propagated plants and trees.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17309681 [PubMed - indexed for MEDLINE]

775: J Exp Bot. 2007;58(6):1281-90. Epub 2007 Feb 17.

Ethylene regulation of fruit softening and cell wall disassembly in Charentais
melon.

Nishiyama K, Guis M, Rose JK, Kubo Y, Bennett KA, Wangjin L, Kato K, Ushijima K,
Nakano R, Inaba A, Bouzayen M, Latche A, Pech JC, Bennett AB.

Graduate School of Natural Science and Technology, Okayama University, Okayama,
700-8530 Japan.

Cell wall disassembly in ripening fruit is highly complex, involving the
dismantling of multiple polysaccharide networks by diverse families of
wall-modifying proteins. While it has been reported in several species that
multiple members of each such family are expressed in the same fruit tissue, it
is not clear whether this reflects functional redundancy, with protein isozymes
from a single enzyme class performing similar roles and contributing equally to
wall degradation, or whether they have discrete functions, with some isoforms
playing a predominant role. Experiments reported here sought to distinguish
between cell wall-related processes in ripening melon that were
softening-associated and softening-independent. Cell wall polysaccharide
depolymerization and the expression of wall metabolism-related genes were
examined in transgenic melon (Cucumis melo var. cantalupensis Naud.) fruit with
suppressed expression of the 1-aminocyclopropane-1-carboxylate oxidase (ACO) gene
and fruits treated with ethylene and 1-methylcyclopropene (1-MCP). Softening was
completely inhibited in the transgenic fruit but was restored by treatment with
exogenous ethylene. Moreover, post-harvest application of 1-MCP after the onset
of ripening completely halted subsequent softening, suggesting that melon fruit
softening is ethylene-dependent. Size exclusion chromatography of cell wall
polysaccharides, from the transgenic fruits, with or without exogenous ethylene,
indicated that the depolymerization of both pectins and xyloglucans was also
ethylene dependent. However, northern analyses of a diverse range of cell
wall-related genes, including those for polygalacturonases, xyloglucan
endotransglucosylase/hydrolases, expansin, and beta-galactosidases, identified
specific genes within single families that could be categorized as
ethylene-dependent, ethylene-independent, or partially ethylene-dependent. These
results support the hypothesis that while individual cell wall-modifying proteins
from each family contribute to cell wall disassembly that accompanies fruit
softening, other closely related family members are regulated in an
ethylene-independent manner and apparently do not directly participate in fruit
softening.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17308329 [PubMed - indexed for MEDLINE]

776: Vet Rec. 2007 Feb 17;160(7):215-8.

Comment in:
Vet Rec. 2007 Mar 10;160(10):347; author reply 347.

Bovine spongiform encephalopathy and the safety of milk from Canadian dairy
cattle.

Tyshenko MG.

McLaughlin Centre for Population Health Risk Assessment, Institute of Population
Health, University of Ottawa, 1 Stewart Street, Ottawa, Ontario, Canada K1N 6N5.

The detection of bovine spongiform encephalopathy (BSE) in beef cattle closed
Canadian beef export markets to 30 countries, including the USA, with devastating
financial losses. The detection and confirmation of the fifth and seventh
BSE-infected animals but first infected dairy cows extended the problem of risk
management to Canadian dairy farmers. As the public are concerned about the
safety not only of beef but also of milk and milk products that may contain
disease-causing prions, this review examines the evidence for the safety of milk
from studies on prions in milk or colostrum and their vertical and lateral
transmission in various animal systems. The evidence indicates that the risk of
contracting new variant Creutzfeldt-Jakob disease through the consumption of milk
is negligible.

Publication Types:
Review

PMID: 17308017 [PubMed - indexed for MEDLINE]

777: Plant Cell. 2007 Feb;19(2):485-94. Epub 2007 Feb 16.

Identification of two protein kinases required for abscisic acid regulation of
seed germination, root growth, and gene expression in Arabidopsis.

Fujii H, Verslues PE, Zhu JK.

Center for Plant Cell Biology, Institute for Integrative Genome Biology and
Department of Botany and Plant Sciences, University of California, Riverside,
California 92521, USA.

Abscisic acid (ABA) is an important phytohormone regulating various plant
processes, including seed germination. Although phosphorylation has been
suggested to be important, the protein kinases required for ABA signaling during
seed germination and seedling growth remain elusive. Here, we show that two
protein kinases, SNF1-RELATED PROTEIN KINASE2.2 (SnRK2.2) and SnRK2.3, control
responses to ABA in seed germination, dormancy, and seedling growth in
Arabidopsis thaliana. A snrk2.2 snrk2.3 double mutant, but not snrk2.2 or snrk2.3
single mutants, showed strong ABA-insensitive phenotypes in seed germination and
root growth inhibition. Changes in seed dormancy and ABA-induced Pro accumulation
consistent with ABA insensitivity were also observed. The snrk2.2 snrk2.3 double
mutant had a greatly reduced level of a 42-kD kinase activity capable of
phosphorylating peptides from ABF (for ABA Response Element Binding Factor)
transcription factors. ABA-induced expression of several genes whose promoters
contain an ABA response element (ABRE) was reduced in snrk2.2 snrk2.3, suggesting
that the mechanism of SnRK2.2 and SnRK2.3 action in ABA signaling involves the
activation of ABRE-driven gene expression through the phosphorylation of ABFs.
Together, these results demonstrate that SnRK2.2 and SnRK2.3 are redundant but
key protein kinases that mediate a major part of ABA signaling in Arabidopsis.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17307925 [PubMed - indexed for MEDLINE]

778: J Biotechnol. 2007 May 1;129(3):539-46. Epub 2007 Jan 21.

Single chain antibody fragments for ocular use produced at high levels in a
commercial wheat variety.

Brereton HM, Chamberlain D, Yang R, Tea M, McNeil S, Coster DJ, Williams KA.

Department of Ophthalmology, Flinders University, Adelaide, SA 5042, Australia.
helen.brereton@flinders.edu.au

We are investigating the use of single chain antibody fragments (scFv) in eye
drops for diagnosis and treatment of eye diseases. For ocular use, recombinant
proteins must be free of bacterial endotoxin that causes inflammation in the eye.
We required a means of generating high yields of scFvs with little endotoxin
contamination. Using microprojectile bombardment we produced transgenic lines of
the commercial wheat variety, Westonia, that express two scFvs that bind to CD4
or CD28 on the surface of rat thymocytes. A high level of expression of active
scFv in the range 50-180 microg/g was measured by quantitative flow cytometry in
crude extracts made from mature seeds. The levels of expression were stable over
four generations of transgenic plants and mature seeds were stored for one year
with little loss of scFv activity. Substantial purification of scFv was achieved
by immobilised metal affinity chromatography. Compared to bacterial extracts,
crude transgenic seed extracts contained only a small amount of endotoxin (150
EU/ml) that will be easily removed by purification. The transgenic wheat lines
express functional scFv at levels comparable to production in bacteria and
promise to be superior to bacteria for production of scFv pharmaceuticals for
ocular use.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17306402 [PubMed - indexed for MEDLINE]

779: Plant Cell Physiol. 2007 Mar;48(3):540-9. Epub 2007 Feb 15.

Characterization of OsPID, the rice ortholog of PINOID, and its possible
involvement in the control of polar auxin transport.

Morita Y, Kyozuka J.

Graduate School of Agriculture and Life Sciences, University of Tokyo, Yayoi,
Bunkyo, Tokyo, 113-0032, Japan.

PINOID, a serine threonine protein kinase in Arabidopsis, controls auxin
distribution through a positive control of subcellular localization of PIN auxin
efflux carriers. Compared with the rapid progress in understanding mechanisms of
auxin action in dicot species, little is known about auxin action in monocot
species. Here, we describe the identification and characterization of OsPID, the
PINOID ortholog of rice. Phylogenetic analysis showed that the rice genome
contains a single PID ortholog, OsPID. Constitutive overexpression of OsPID
caused a variety of abnormalities, such as delay of adventitious root
development, curled growth of shoots and agravitropism. Abnormalities observed in
the plants that overexpress OsPID could be phenocopied by treatment with an
inhibitor of active polar transport of auxin, indicating that OsPID could be
involved in the control of polar auxin transport in rice. Analysis of OsPID mRNA
distribution showed a complex pattern in shoot meristems, indicating that it
probably plays a role in the pattern formation and organogenesis in the rice
shoot.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17303594 [PubMed - indexed for MEDLINE]

780: J Am Vet Med Assoc. 2007 Feb 15;230(4):464-6, 468.

Animal clones in the food supply.

Burns K.

Publication Types:
News

PMID: 17302542 [PubMed - indexed for MEDLINE]

781: Minerva Pediatr. 2007 Feb;59(1):35-41.

Rice protein-based infant formula: current status and future development.

Koo WW, Lasekan JB.

The Carman and Ann Adams, Department of Pediatrics, Wayne State University,
Hutzel Women's Hospital, Detroit, MI 48201, USA. wkoo@wayne.edu

Rice is the world's leading staple cereal food and is the major source of protein
for many parts of the world. Rice is among the first solid foods fed to infants
in many cultures, in part because of its hypoallergenicity from lack of gluten.
Nutritional quality of rice protein compares favorably with other cereal proteins
including wheat, oat and barley. It is rich in methionine and cystine, although
as is the case for other cereals, it is an incomplete protein source for human
infants with lysine and threonine being the primary limiting amino acids.
Fortification of rice proteins with these two limiting amino acids improves its
protein quality. Rice protein-based infant formulas (RPF) were initially based on
high protein rice flours, but more recently are based on rice protein
concentrates, isolates or hydrolysates, fortified with lysine and threonine.
Hypoallergenicity efficacy, particularly for hydrolyzed rice protein-based
formulas, has been reported, and limited data indicated that rice protein based
infant formula may provide potentially adequate alternative if standard milk- or
soy protein-based formulas are not tolerated. Unlike the rice-protein based
infant formula, rice beverage formulas made from rice flour are nutritionally
inadequate for infants. Reports have indicated stunted growth in infants/children
fed rice beverage formulas. Future development for the RPF include those based on
genetically improved rice with high lysine and threonine content, supplementation
with appropriate mineral and fat blend, and long-term clinical studies in infants
to confirm its efficacy and safety.

PMID: 17301723 [PubMed - indexed for MEDLINE]

782: J Biosci. 2006 Dec;31(5):617-27.

Combinative effects of a bacterial type-III effector and a biocontrol bacterium
on rice growth and disease resistance.

Ren H, Gu G, Long J, Yin Q, Wu T, Song T, Zhang S, Chen Z, Dong H.

Key Laboratory of Monitoring and Management of Plant Pathogens and Insect Pests,
Ministry of Agriculture of China, and Department of Plant Pathology, Nanjing
Agricultural University, 1 Weigang, Nanjing, 210095, China.

Expression of HpaG(Xoo), a bacterial type-III effector, in transgenic plants
induces disease resistance. Resistance also can be elicited by biocontrol
bacteria. In both cases, plant growth is often promoted. Here we address whether
biocontrol bacteria and HpaG(Xoo) can act together to provide better results in
crop improvement. We studied effects of Pseudomonas cepacia on the rice variety
R109 and the hpaG(Xoo)-expressing rice line HER1. Compared to R109, HER1 showed
increased growth, grain yield, and defense responses toward diseases and salinity
stress. Colonization of roots by P. cepacia caused 20% and 13% increase, in
contrast to controls, in root growth of R109 and HER1. Growth of leaves and stems
also increased in R109 but that of HER1 was inhibited. When P. cepacia
colonization was subsequent to plant inoculation with Rhizoctonia solani, a
pathogen that causes sheath blight, the disease was less severe than controls in
both R109 and HER1; HER1, nevertheless, was more resistant, suggesting that P.
cepacia and HpaG(Xoo) cooperate in inducing disease resistance. Several genes
that critically regulate growth and defense behaved differentially in HER1 and
R109 while responding to P. cepacia. In R109 leaves, the OsARF1 gene, which
regulates plant growth, was expressed in consistence with growth promotion by P.
cepacia. Inversely, OsARF1 expression was coincident with inhibition in growth of
HER1 leaves. In both plants, the expression of OsEXP1, which encodes an expansin
protein involved in plant growth,was concomitant with growth promotion in leaves
instead of roots,in response to P. cepacia . We also studied OsMAPK, a gene that
encodes a mitogen-activated protein kinase and controls defense responses toward
salinity and infection by pathogens in rice. In response to P. cepacia, an early
expression of OsMAPK was coincident with R109 resistance to the disease, while
HER1 expressed the gene similarly whether P. cepacia was present or not.
Evidently, P. cepacia and G(Xoo)-gene mediated resistance may act differently in
rice growth and resistance. Whereas combinative effects of P. cepacia and
HpaG(Xoo) in disease resistance have a great potential in agricultural use, it is
interesting to study mechanisms that underlie interactions involving biocontrol
bacteria, type-III effectors and pathogens.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17301500 [PubMed - indexed for MEDLINE]

783: J Agric Food Chem. 2007 Feb 21;55(4):1264-73.

Development of a real-time PCR method based on duplo target plasmids for
determining an unexpected genetically modified soybean intermix with feed
components.

Dalla Costa L, Martinelli L.

IASMA Research Center, Genetics and Molecular Biology Department, Via E. Mach 1,
38010 San Michele all'Adige (TN), Italy.

The occurrence of intermixing, especially that resulting from genetically
modified (GM) species, is increasingly becoming a problem in the delicate chain
of feed and food quality control. Thus, a strategy is needed for precisely
quantifying the presence of intermixing. An analytical assay based on real-time
PCR has been developed; it can ascertain the extent of unexpected intermixing of
GM soybean with maize meal. Three soybean-maize mix levels, with soybean intermix
percentages of, respectively, 0.1, 0.5, and 1%, were prepared to simulate samples
containing traces of soybean. As calibrator standards, ad hoc multiple-target
pGEM-T plasmids containing soybean and maize reference genes in a 1:1 ratio were
constructed. Four different maize endogenous genes, alcohol dehydrogenase 1
(adh1), high-mobility group protein a (hmga), invertase 1 (ivr1), and zein
(zein), were assessed, each combined with the soybean endogenous lectin 1 (lect1)
gene. Plasmids containing adh1-lect1 and zein-lect1 genes were found to be the
most reliable calibration systems for this analysis, providing precise and
accurate quantification results. Measuring the percentage of GM soybean
intermixing makes it possible to calculate the actual transgenic component of the
total sample.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17300150 [PubMed - indexed for MEDLINE]

784: J Agric Food Chem. 2007 Feb 21;55(4):1071-6.

First application of a microsphere-based immunoassay to the detection of
genetically modified organisms (GMOs): quantification of Cry1Ab protein in
genetically modified maize.

Fantozzi A, Ermolli M, Marini M, Scotti D, Balla B, Querci M, Langrell SR, Van
den Eede G.

Biotechnology and GMOs Unit, Institute for Health and Consumer Protection,
DG-Joint Research Centre, European Commission, I-2120 Ispra (Va), Italy.

An innovative covalent microsphere immunoassay, based on the usage of fluorescent
beads coupled to a specific antibody, was developed for the quantification of the
endotoxin Cry1Ab present in MON810 and Bt11 genetically modified (GM) maize
lines. In particular, a specific protocol was developed to assess the presence of
Cry1Ab in a very broad range of GM maize concentrations, from 0.1 to 100% [weight
of genetically modified organism (GMO)/weight]. Test linearity was achieved in
the range of values from 0.1 to 3%, whereas fluorescence signal increased
following a nonlinear model, reaching a plateau at 25%. The limits of detection
and quantification were equal to 0.018 and 0.054%, respectively. The present
study describes the first application of quantitative high-throughput
immunoassays in GMO analysis.

PMID: 17300145 [PubMed - indexed for MEDLINE]

785: Riv Biol. 2006 Sep-Dec;99(3):381-94.

The impact of GMOs on poor countries: a threat to the achievement of the
Millennium Development Goals?

Francescon S.

United Nations Millennium Campaign.

The first of the Millennium Development Goals - halve the proportion of people
who suffer from hunger by 2015 - is essential for eradicating poverty, as most of
the poor live in rural areas.The role of agriculture is, therefore, key to the
fight against poverty.Nevertheless, over the last years rich countries diminished
their official development assistance for agricultural development and some of
them proposed and pushed for a new model of agriculture based on biotechnology.
Such a new model of agriculture is presented by its supporters as a means to
contribute to the elimination of poverty, as it intends to maximise the crop
production.However, it does not take into consideration that policies fighting
hunger: need a more comprehensive approach; must take into consideration
socio-economic and environmental peculiarities, especially local needs and
traditional knowledge and practices.Genetically modified technology goes against
these basic requirements, as it is designed to suit multinational enterprises in
the North.When drafting development policies, rich and poor countries must bear
in mind that the framework of the Millennium Development Goals, to which 189
Nations committed, requires a coherent approach to empower the poor, especially
women, and promote traditional knowledge of indigenous people and local
communities, as well as ensuring environmental sustainability.The fight to
poverty and hunger will not be won and people will still go hungry if the
fundamental causes of hunger and food insecurity are not tackled, whereas
genetically modified technology is not based on this assumption.

Publication Types:
Review

PMID: 17299696 [PubMed - indexed for MEDLINE]

786: J Fish Dis. 2007 Feb;30(2):65-79.

Histological, digestive, metabolic, hormonal and some immune factor responses in
Atlantic salmon, Salmo salar L., fed genetically modified soybeans.

Bakke-McKellep AM, Koppang EO, Gunnes G, Sanden M, Hemre GI, Landsverk T,
Krogdahl A.

Aquaculture Protein Centre, CoE, Norway. anne.mckellep@veths.no

The paper reports the second and final part of an experiment aiming to study
physiological and health-related effects of genetically modified (GM) soybean
meal (SBM) type Roundup Ready soybean (RRS) in diets for post-smolt Atlantic
salmon. For 3 months salmon were fed diets containing 172 g kg(-1) full-fat SBM
from RRS (GM-soy) or an unmodified, non-isogenic line (nGM-soy), or a reference
diet with fishmeal as the sole protein source (FM). Slight differences in
anti-nutrient levels were observed between the GM and nGM-soy. Histological
changes were observed only in the distal intestine of the soy-fed fish. The
incidence of moderate inflammation was higher in the GM-soy group (9 of 10
sampled fish) compared with the nGM-soy group (7 of 10). However, no differences
in the concomitant decreases in activities of digestive enzymes located in the
brush border (leucine aminopeptidase and maltase) and apical cytoplasm (acid
phosphatase) of enterocytes or in the number of major histocompatibility complex
class II+ cells, lysozyme activity, or total IgM of the distal intestine were
observed. GM compared with nGM-soy fed fish had higher head kidney lysozyme
(11,856 vs. 10,456 units g(-1) tissue) and a tendency towards higher acid
phosphatase (0.45 vs. 0.39 micromol h(-1) kg(-1) body mass in whole tissue)
activities, respectively. Plasma insulin and thyroxin levels, and hepatic
fructose-1,6-bisphosphatase and ethoxyresorufin-O-deethylase activities were not
significantly affected. It is not possible, however, to conclude whether the
differences in responses to GM-soy were due to the genetic modification or to
differences in soy cultivars in the soy-containing diets. Results from studies
using non-modified, parental line soybeans as the control group are necessary to
evaluate whether genetic modification of soybeans in diets poses any risk to
farmed Atlantic salmon.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17298562 [PubMed - indexed for MEDLINE]

787: Z Naturforsch [C]. 2006 Nov-Dec;61(11-12):833-9.

Effect of drought stress at supraoptimal temperature on polyamine concentrations
in transgenic soybean with increased proline levels.

Simon-Sarkadi L, Kocsy G, Vö¡rhegyi A, Galiba G, de Ronde JA.

Department of Biochemistry and Food Technology, Budapest University of Technology
and Economics, H-1521 Budapest, P.O.B. 91, Hungary. sarkadi@mail.bme.hu

The effect of drought stress at supraoptimal temperature on free proline and
polyamine levels was compared in wild type and transgenic soybean (Glycine max
cv. Ibis) plants having increased proline levels. Since glutamate and arginine
are precursors of both proline and polyamines, it was assumed that the genetic
manipulation of proline levels would also affect the polyamine levels. The
proline and spermine concentrations increased, while the putrescine concentration
generally decreased or did not change after the treatments in both genotypes.
Following drought higher proline and lower spermine levels were detected in the
transgenic plants compared to the wild type ones, which could be explained by the
increased use of their common precursors for proline biosynthesis in the
transgenic plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17294695 [PubMed - indexed for MEDLINE]

788: Plant Physiol. 2007 Apr;143(4):1739-51. Epub 2007 Feb 9.

Overexpression of an R1R2R3 MYB gene, OsMYB3R-2, increases tolerance to freezing,
drought, and salt stress in transgenic Arabidopsis.

Dai X, Xu Y, Ma Q, Xu W, Wang T, Xue Y, Chong K.

Research Center for Molecular and Developmental Biology, Key Laboratory of
Photosynthesis and Environmental Molecular Physiology, Institute of Botany,
Chinese Academy of Sciences, Beijing 100093, China.

We used a cDNA microarray approach to monitor the expression profile of rice
(Oryza sativa) under cold stress and identified 328 cold-regulated genes.
Thirteen such genes encoding MYB, homeodomain, and zinc finger proteins with
unknown functions showed a significant change in expression under 72-h cold
stress. Among them, OsMYB3R-2 was selected for further study. Unlike most plant
R2R3 MYB transcription factors, OsMYB3R-2 has three imperfect repeats in the
DNA-binding domain, the same as in animal c-MYB proteins. Expression of OsMYB3R-2
was induced by cold, drought, and salt stress. The Arabidopsis (Arabidopsis
thaliana) transgenic plants overexpressing OsMYB3R-2 showed increased tolerance
to cold, drought, and salt stress, and the seed germination of transgenic plants
was more tolerant to abscisic acid or NaCl than that of wild type. The expression
of some clod-related genes, such as dehydration-responsive element-binding
protein 2A, COR15a, and RCI2A, was increased to a higher level in
OsMYB3R-2-overexpressing plants than in wild type. These results suggest that
OsMYB3R-2 acts as a master switch in stress tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17293435 [PubMed - indexed for MEDLINE]

789: Plant Cell Physiol. 2007 Mar;48(3):523-39. Epub 2007 Feb 9.

Overexpression of a type-A response regulator alters rice morphology and
cytokinin metabolism.

Hirose N, Makita N, Kojima M, Kamada-Nobusada T, Sakakibara H.

RIKEN Plant Science Center, 1-7-22, Suehiro, Tsurumi, Yokohama, 230-0045, Japan.

Genome-wide analyses of rice (Oryza sativa L.) cytokinin (CK)-responsive genes
using the Affymetrix GeneChip(R) rice genome array were conducted to define the
spectrum of genes subject to regulation by CK in monocotyledonous plants.
Application of trans-zeatin modulated the expression of a wide variety of genes
including those involved in hormone signaling and metabolism, transcriptional
regulation, macronutrient transport and protein synthesis. To understand further
the function of CK in rice plants, we examined the effects of in planta
manipulation of a putative CK signaling factor on morphology, CK metabolism and
expression of CK-responsive genes. Overexpression of the CK-inducible type-A
response regulator OsRR6 abolished shoot regeneration, suggesting that OsRR6 acts
as a negative regulator of CK signaling. Transgenic lines overexpressing OsRR6
(OsRR6-ox) had dwarf phenotypes with poorly developed root systems and panicles.
Increased content of trans-zeatin-type CKs in OsRR6-ox lines indicates that
homeostatic control of CK levels is regulated by OsRR6 signaling. Expression of
genes encoding CK oxidase/dehydrogenase decreased in OsRR6-ox plants, possibly
accounting for elevated CK levels in transgenic lines. Expression of a number of
stress response genes was also altered in OsRR6-ox plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17293362 [PubMed - indexed for MEDLINE]

790: Glycobiology. 2007 Jun;17(6):600-19. Epub 2007 Feb 9.

N-glycans of recombinant human acid alpha-glucosidase expressed in the milk of
transgenic rabbits.

Jongen SP, Gerwig GJ, Leeflang BR, Koles K, Mannesse ML, van Berkel PH, Pieper
FR, Kroos MA, Reuser AJ, Zhou Q, Jin X, Zhang K, Edmunds T, Kamerling JP.

Bijvoet Center for Biomolecular Research, Department of Bio-Organic Chemistry,
Utrecht University, Padualaan 8, NL-3584 CH Utrecht, The Netherlands.

Pompe disease is a lysosomal glycogen storage disorder characterized by acid
alpha-glucosidase (GAA) deficiency. More than 110 different pathogenic mutations
in the gene encoding GAA have been observed. Patients with this disease are being
treated by intravenous injection of recombinant forms of the enzyme. Focusing on
recombinant approaches to produce the enzyme means that specific attention has to
be paid to the generated glycosylation patterns. Here, human GAA was expressed in
the mammary gland of transgenic rabbits. The N-linked glycans of recombinant
human GAA (rhAGLU), isolated from the rabbit milk, were released by
peptide-N(4)-(N-acetyl-beta-glucosaminyl)asparagine amidase F. The N-glycan pool
was fractionated and purified into individual components by a combination of
anion-exchange, normal-phase, and Sambucus nigra agglutinin-affinity
chromatography. The structures of the components were analyzed by 500 MHz
one-dimensional and 600 MHz cryo two-dimensional (total correlation spectroscopy
[TOCSY] nuclear Overhauser enhancement spectroscopy) (1)H nuclear magnetic
resonance spectroscopy, combined with two-dimensional (31)P-filtered (1)H-(1)H
TOCSY spectroscopy, matrix-assisted laser desorption ionization time-of-flight
mass spectrometry, and high-performance liquid chromatography (HPLC)-profiling of
2-aminobenzamide-labeled glycans combined with exoglycosidase digestions. The
recombinant rabbit glycoprotein contained a broad array of different N-glycans,
comprising oligomannose-, hybrid-, and complex-type structures. Part of the
oligomannose-type glycans showed the presence of phospho-diester-bridged
N-acetylglucosamine. For the complex-type glycans (partially)
(alpha2-6)-sialylated (nearly only N-acetylneuraminic acid) diantennary
structures were found; part of the structures were (alpha1-6)-core-fucosylated or
(alpha1-3)-fucosylated in the upper antenna (Lewis x). Using HPLC-mass
spectrometry of glycopeptides, information was generated with respect to the
site-specific location of the various glycans.

PMID: 17293352 [PubMed - indexed for MEDLINE]

791: J Allergy Clin Immunol. 2007 Apr;119(4):1013-6. Epub 2007 Feb 9.

Vaccination with genetically modified birch pollen allergens: immune and clinical
effects on oral allergy syndrome.

Niederberger V, Reisinger J, Valent P, Krauth MT, Pauli G, van Hage M, Cromwell
O, Horak F, Valenta R.

Publication Types:
Comparative Study
Letter
Multicenter Study
Randomized Controlled Trial
Research Support, Non-U.S. Gov't

PMID: 17292956 [PubMed - indexed for MEDLINE]

792: FEBS Lett. 2007 Mar 6;581(5):898-904. Epub 2007 Feb 5.

Oral secretions from herbivorous lepidopteran larvae exhibit ion channel-forming
activities.

Maischak H, Grigoriev PA, Vogel H, Boland W, Mithö¶fer A.

Max-Planck-Institut fö¼r Chemische Okologie, Hans-Knö¶ll-Str. 8, D-07745 Jena,
Germany.

Feeding insects introduce oral secretions (OS) into the wounded tissue of the
attacked plant. Various OS-derived molecules must be involved in subsequent
processes including the induction of plant defence reactions. Using the planar
lipid bilayer membrane technique, isolated OS were analyzed with respect to their
membrane activities. Transmembrane ion fluxes were generated by OS of eight
different lepidopteran larvae, all of which form comparable ion channels in
artificial membranes. Currents were characterized by long lasting open times and
conductivities from 250pS up to 1100pS. Channels formed by Spodoptera exigua
secretions showed a preference for cations over anions. OS also induced a
transient increase of the cytosolic calcium concentration in soybean cells,
determined by the aequorin technique. Known compounds of the OS,
fatty-acid-glutamine conjugates, also interfered with the membrane but were
unable to form stable channels. Since ion fluxes and depolarization are early
responses upon insect feeding, OS-derived components may be involved in the
elicitation process by direct interaction with the plant membranes.

Publication Types:
In Vitro
Research Support, Non-U.S. Gov't

PMID: 17292890 [PubMed - indexed for MEDLINE]

793: Plant J. 2007 Mar;49(5):800-9. Epub 2007 Feb 9.

Clp protease controls chlorophyll b synthesis by regulating the level of
chlorophyllide a oxygenase.

Nakagawara E, Sakuraba Y, Yamasato A, Tanaka R, Tanaka A.

Institute of Low Temperature Science, Hokkaido University, N19 W8 Kita-ku,
Sapporo, Japan.

Chlorophyll b is one of the major light-harvesting pigments in green plants and
it is essential for optimal light harvesting. Chlorophyll b is synthesized from
chlorophyll a by chlorophyllide a oxygenase (CAO) which consists of A, B and C
domains. Previously, we demonstrated that the C domain alone has a catalytic
function, while the A and B domains control the level of CAO protein in response
to chlorophyll b accumulation. We hypothesized that the accumulation of
chlorophyll b triggers the proteolytic degradation of CAO. In this study, in
order to gain further insight into this regulatory mechanism we screened for
mutants that have defects in the control of CAO accumulation. Seeds from a
transgenic line of Arabidopsis which overexpressed a CAO-GFP fusion were
mutagenized and their progenies were screened by laser-scanning confocal
microscopy for mutants showing an elevated level of GFP fluorescence. One
particular mutant (dca1) exhibited stronger GFP fluorescence and accumulated a
GFP-CAO fusion protein at a higher level. Concomitantly, the chlorophyll a to b
ratio decreased in this mutant. The mutation in the dca1 mutant was mapped to the
ClpC1 gene, thereby indicating that a chloroplast Clp protease is involved in
regulating chlorophyll b biosynthesis through the destabilization of CAO protein
in response to the accumulation of chlorophyll b.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17291312 [PubMed - indexed for MEDLINE]

794: Ukr Biokhim Zh. 2006 Sep-Oct;78(5):70-80.

[Changed ratios of some pigments of photosynthesis in Nicotiana tabacum L.
induced by exogenic DNA]

[Article in Ukrainian]

Katsan VA, Potopal's'kyä­ AI.

The technology of obtaining the tobacco plants possessing the hereditary changes
in the photosynthesis pigments accumulation during development using exogenous
DNA has been elaborated. The plants possessing changes in proportion of
chlorophylls content in the leave tissues, inherited elevated carotenoid content
and altered proportion of carotene and xanthophylls, luteine and violaxanthine
have been obtained by action of the salt-tolerant nightshade (Solanum nigrum L.)
DNA and the DNAs of pCAMVNEO, pTi8628 plasmids on the tobacco cultivar
Krupnolystny 20 (Large-leaf) germinating seeds. These plants have simultaneously
the useful features--accelerated development, early blooming phenotype and higher
productivity. Possible mechanisms emphasized such inherited biochemical changes
have been discussed.

Publication Types:
English Abstract

PMID: 17290784 [PubMed - indexed for MEDLINE]

795: J Agric Food Chem. 2007 Mar 7;55(5):1900-4. Epub 2007 Feb 9.

Biodegradation of Cry1Ab protein from Bt transgenic rice in aerobic and flooded
paddy soils.

Wang H, Ye Q, Gan J, Wu L.

Institute of Nuclear Agricultural Sciences, Zhejiang University, Hangzhou 310029,
China.

Degradation of Cry1Ab protein from Bt transgenic rice was examined under both
aerobic and flooded conditions in five paddy soils and in aqueous solutions. The
hydrolysis rate of Cry1Ab protein in aqueous solutions was correlated inversely
with the solution pH in the range of 4.0 to 8.0, and positively with the initial
concentration of Cry1Ab protein. Rapid degradation of Cry1Ab protein occurred in
paddy soils under aerobic conditions, with half-lives ranging from 19.6 to 41.3
d. The degradation was mostly biotic and not related to any specific soil
property. Degradation of the Cry1Ab protein was significantly prolonged under
flooded conditions compared with aerobic conditions, with half-lives extended to
45.9 to 141 d. These results suggest that the toxin protein, when introduced into
a paddy field upon harvest, will probably undergo rapid removal after the field
is drained and exposed to aerobic conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17288444 [PubMed - indexed for MEDLINE]

796: Theor Appl Genet. 2007 May;114(7):1141-50. Epub 2007 Feb 8.

Negative effect of chromosome 1A on dough strength shown by modification of 1D
addition in durum wheat (Triticum durum).

Garg M, Dhaliwal HS, Chhuneja P, Kumar D, Dou QW, Tanaka H, Elamein HM, Tsujimoto
H.

Laboratory of Plant Genetics and Breeding Science, Faculty of Agriculture,
Tottori University, 4-101 Minami, Koyama, Tottori, 680-8553, Japan.

A monosomic addition line of Aegilops tauschii chromosome 1D in Triticum durum
cv. PBW114 was produced in 1990. This line was self-pollinated and maintained for
several generations while following the presence of chromosome 1D carrying the
gene for red glume color. Cytological analysis indicated that two of the three
derivative lines had substitution of chromosome 1D for 1A and another had
substitution of chromosome 1D for 1B. One of these lines carried a pair of small
chromosomes in addition to the 1D chromosome. Sodium dodecyl
sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the derived lines showed
the presence of high-molecular-weight (HMW) glutenin encoded by the Glu-D1 locus.
The small chromosome found in one of the lines had nearly regular pairing and
transmission to daughter nuclei. Fluorescent in situ hybridization (FISH) and
analysis of molecular markers indicated that the small chromosome was derived
from the short arm of chromosome 1A and carried the Glu-A3 locus. Microsatellite
mapping based on the deletion bin map revealed that the small chromosome had
terminal deletions on both the terminal and centromeric sides. The line with the
small chromosome showed improvement of the sodium dodecyl sulfate
(SDS)-sedimentation value as compared to parent durum. However, the increase in
SDS-sedimentation value was more significant in the substitution line of
chromosome 1D for 1A without the small chromosome. These facts suggest a negative
effect of the Glu-A3 locus on dough strength. The sequence of the Glu-D1 locus
from these lines showed that the HMW glutenin subunits were Ae. tauschii specific
2(t) + T2, which were previously found to be associated with poor rheological
properties and bread loaf volume in synthetic hexaploid wheat by other workers.
Thus, the significant improvement in the SDS-sedimentation value of the
substitution line of 1D for 1A suggests that the absence of the negative effect
of chromosome 1A on quality is more important than the presence of Glu-D1 of Ae.
tauschii.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17287973 [PubMed - indexed for MEDLINE]

797: Plant Mol Biol. 2007 Apr;63(6):815-32. Epub 2007 Feb 8.

Transgenic cotton over-producing spinach sucrose phosphate synthase showed
enhanced leaf sucrose synthesis and improved fiber quality under controlled
environmental conditions.

Haigler CH, Singh B, Zhang D, Hwang S, Wu C, Cai WX, Hozain M, Kang W, Kiedaisch
B, Strauss RE, Hequet EF, Wyatt BG, Jividen GM, Holaday AS.

Department of Crop Science and Department of Plant Biology, North Carolina State
University, Raleigh, NC 27695-7620, USA. candace_haigler@ncsu.edu

Prior data indicated that enhanced availability of sucrose, a major product of
photosynthesis in source leaves and the carbon source for secondary wall
cellulose synthesis in fiber sinks, might improve fiber quality under abiotic
stress conditions. To test this hypothesis, a family of transgenic cotton plants
(Gossypium hirsutum cv. Coker 312 elite) was produced that over-expressed spinach
sucrose-phosphate synthase (SPS) because of its role in regulation of sucrose
synthesis in photosynthetic and heterotrophic tissues. A family of 12 independent
transgenic lines was characterized in terms of foreign gene insertion, expression
of spinach SPS, production of spinach SPS protein, and development of enhanced
extractable V (max) SPS activity in leaf and fiber. Lines with the highest V
(max) SPS activity were further characterized in terms of carbon partitioning and
fiber quality compared to wild-type and transgenic null controls. Leaves of
transgenic SPS over-expressing lines showed higher sucrose:starch ratio and
partitioning of (14)C to sucrose in preference to starch. In two growth chamber
experiments with cool nights, ambient CO(2) concentration, and limited light
below the canopy, the transgenic line with the highest SPS activity in leaf and
fiber had higher fiber micronaire and maturity ratio associated with greater
thickness of the cellulosic secondary wall.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17287885 [PubMed - indexed for MEDLINE]

798: Nat Biotechnol. 2007 Feb;25(2):169-70.

Comment on:
Nat Biotechnol. 2006 Jan;24(1):23-5.

Model for tuning GMO detection in seed and grain.

Macarthur R, Murray AW, Allnutt TR, Deppe C, Hird HJ, Kerins GM, Blackburn J,
Brown J, Stones R, Hugo S.

Publication Types:
Comment
Letter
Research Support, Non-U.S. Gov't

PMID: 17287745 [PubMed - indexed for MEDLINE]

799: Nat Biotechnol. 2007 Feb;25(2):166-7; discussion 167.

Comment on:
Nat Biotechnol. 2006 Oct;24(10):1191-3.

The fit between organic and pharma crops in North Carolina.

Williams CG.

Publication Types:
Comment
Letter
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17287742 [PubMed - indexed for MEDLINE]

800: Nat Biotechnol. 2007 Feb;25(2):165; discussion 165-6.

Comment on:
Nat Biotechnol. 2006 Oct;24(10):1177.

Blame factory farming, not organic food.

Holdrege C.

Publication Types:
Comment
Letter

PMID: 17287741 [PubMed - indexed for MEDLINE]

801: Biotechnol Prog. 2007 Mar-Apr;23(2):297-301. Epub 2007 Feb 8.

Comparative evaluation of different DNA extraction procedures from food samples.

Di Bernardo G, Del Gaudio S, Galderisi U, Cascino A, Cipollaro M.

Department of Experimental Medicine, Section of Biotechnologies and Molecular
Biology, and CRISCEB, 2nd University of Naples, Via Costantinopoli 16, 80138
Naples, Italy.

Five methodologies for extracting DNA from food samples are described. The food
products analyzed are from either soybean or maize. They were selected on the
basis of the mechanical, thermal, and chemical treatments that they had been
subjected to during industrial processing. DNA preparations were evaluated for
purity, yield, and average fragment size. Two endogenous genes, soybean lectin
gene and alcohol dehydrogenase gene (adh1), were used to assess the degree of DNA
degradation at different stages of the transformation chain. The goal of this
study was to determine the role that extraction methods play in DNA amplification
in order to select the best protocol for a food sample. This comparative
evaluation can be specifically useful for detection of genetically modified
ingredients in a variety of food matrices.

Publication Types:
Comparative Study
Evaluation Studies
Research Support, Non-U.S. Gov't
Review

PMID: 17286386 [PubMed - indexed for MEDLINE]

802: Biotechnol J. 2007 Apr;2(4):486-91.

Bovine fetal microchimerism in normal and embryo transfer pregnancies and its
implications for biotechnology applications in cattle.

Turin L, Invernizzi P, Woodcock M, Grati FR, Riva F, Tribbioli G, Laible G.

Dipartimento di Patologia Animale, Igiene e Sanita' Pubblica Veterinaria,
Universita' degli Studi di Milano, Milano, Italy. lauretta.turin@unimi.it

Fetal cells and DNA have been detected in the maternal circulation during and
after pregnancy in a few mammalian species. The incidence of similar
microchimerism in cattle could have repercussion for the application of modern
biotechnologies such as the transfer of transgenic embryos. To determine if
feto-maternal leakage can occur in pregnant cows, we have analyzed maternal blood
samples for the presence of fetal DNA during gestation and post-partum periods. Y
chromosome-specific DNA was detected in up to 73% of blood samples from naturally
mated heifers carrying conventional bull calves and a transgene-specific sequence
in up to 50% of recipient cows carrying transgenic fetuses. These findings
document for the first time that transplacental leakage of fetal DNA into the
maternal circulation can occur in cattle despite the epitheliochorial placenta of
ruminants, with potential implications for the utilization of recipient cows in
the food chain.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17285678 [PubMed - indexed for MEDLINE]

803: Plant Cell Physiol. 2007 Mar;48(3):471-83. Epub 2007 Feb 6.

GASA4, one of the 14-member Arabidopsis GASA family of small polypeptides,
regulates flowering and seed development.

Roxrud I, Lid SE, Fletcher JC, Schmidt ED, Opsahl-Sorteberg HG.

Genetwister Technologies BV, PO Box 193, NL-6700 AD Wageningen, The Netherlands.

Members of the plant-specific gibberellic acid-stimulated Arabidopsis (GASA) gene
family play roles in hormone response, defense and development. We have
identified six new Arabidopsis GASA genes, bringing the total number of family
members to 14. Here we show that these genes all encode small polypeptides that
share the common structural features of an N-terminal putative signal sequence, a
highly divergent intermediate region and a conserved 60 amino acid C-terminal
domain containing 12 conserved cysteine residues. Analysis of promoter::GUS
(beta-glucuronidase) transgenic plants representing six different GASA loci
reveals that the promoters are activated in a variety of stage- and
tissue-specific patterns during development, indicating that the GASA genes are
involved in diverse processes. Characterization of GASA4 shows that the promoter
is active in the shoot apex region, developing flowers and developing embryos.
Phenotypic analyses of GASA4 loss-of-function and gain-of-function lines indicate
that GASA4 regulates floral meristem identity and also positively affects both
seed size and total seed yield.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17284469 [PubMed - indexed for MEDLINE]

804: Cancer Sci. 2007 Apr;98(4):471-7. Epub 2007 Jan 31.

Protective effects of citrus nobiletin and auraptene in transgenic rats
developing adenocarcinoma of the prostate (TRAP) and human prostate carcinoma
cells.

Tang M, Ogawa K, Asamoto M, Hokaiwado N, Seeni A, Suzuki S, Takahashi S, Tanaka
T, Ichikawa K, Shirai T.

Department of Experimental Pathology and Tumor Biology, Nagoya City University
Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya
467-8601, Japan.

Dietary phytochemicals, including nobiletin and auraptene, have been shown to
exert inhibiting effects in several chemically induced carcinogenesis models. We
here investigated the influence of nobiletin and auraptene on prostate
carcinogenesis using transgenic rats developing adenocarcinoma of the prostate
(TRAP) bearing the SV40 T antigen transgene under control of the probasin
promoter and human prostate cancer cells. Starting at 5 weeks of age, male TRAP
rats received powder diet containing 500 p.p.m. nobiletin or auraptene, or the
basal diet for 15 weeks and then were sacrificed for analysis of serum
testosterone levels and histological changes. The body and relative prostate
weights and serum testosterone levels did not differ among the groups. Since all
animals developed prostate carcinomas, these were semiquantitatively measured and
expressed as relative areas of prostate epithelial cells. Nobiletin caused
significant reduction in the ventral (P<0.01), lateral (P<0.001) and dorsal
(P<0.05) prostate lobes, while decreasing high grade lesions (P<0.05) in the
ventral and lateral lobes. Feeding of auraptene also effectively reduced the
epithelial component (P<0.05) and high grade lesions (P<0.05), in the lateral
prostate. A further experiment demonstrated that growth of androgen sensitive
LNCaP and androgen insensitive DU145 and PC3 human prostate cancer cells, was
suppressed by both nobiletin and to a lesser extent auraptene in a dose-dependent
manner, with significant increase in apoptosis. In conclusion, these compounds,
particularly nobiletin, may be valuable for prostate cancer prevention.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17284254 [PubMed - indexed for MEDLINE]

805: Theor Appl Genet. 2007 May;114(7):1203-9. Epub 2007 Feb 6.

Improved yielding and reduced puffiness under extreme temperatures induced by
fruit-specific expression of rolB in processing tomatoes.

Shabtai S, Salts Y, Kaluzky G, Barg R.

Institute of Plant Sciences, The Volcani Center, ARO, P.O. Box 6, Bet Dagan,
50250, Israel.

Tomato fruit production is severely hampered by both extremely high and low
temperatures, mainly due to impaired microsporogenesis and pollination under
these conditions. Even mild temperature stress, leading to partial damage to
pollen viability can result in the production of under-fertilized puffy fruits of
poor quality, while severe stress can abolish fruit set completely. Genetic or
transgenic parthenocarpy that enables fertilization-independent fruit development
offers a solution for tomato yielding under conditions unfavorable for pollen
production and/or fertilization. A transgenic processing tomato UC82 line,
expressing rolB specifically during early stages of fruit development was
compared to the parental line with respect to yield and fruit quality under
extreme temperatures. Under both high and low temperatures the transgenic line
performed significantly better than the parental line. Its yield was
significantly higher mainly due to a higher number of fruits that did develop,
and also because of increased fruit weight. While the UC82 fruits developed under
high temperatures were very puffy and severely malformed, the transgenic fruits
maintained improved jelly fill and were of smooth and regular shape.
Interestingly, under high temperatures the improved jelly fill in the transgenic
line was accompanied by a higher number of seeds, suggesting that not only the
developing seeds promote development of the placental tissue but also that
proliferation of this tissue supports better seed development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17279365 [PubMed - indexed for MEDLINE]

806: Immunol Allergy Clin North Am. 2007 Feb;27(1):129-39.

A rice-based edible vaccine expressing multiple T-cell epitopes to induce oral
tolerance and inhibit allergy.

Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.
takaiwa@nias.affrc.go.jp

Plant pollens are the most common cause of seasonal allergic disease. The number
of patients undergoing treatment for allergies to the pollen of Japanese cedar
(major antigens, Cry j 1 and Cry j 2) has increased steadily each year. A rice
seed-based edible vaccine has been shown to be effective for treating Japanese
cedar pollinosis. Rice seeds containing the major T-cell epitopes derived from
cedar pollen allergens were orally administrated to mice before systemic
challenge with total pollen protein. Mucosal immune tolerance leading to a
reduction of allergen-specific IgE, T-cell proliferative reactions, and histamine
were induced, resulting in suppression of allergy-specific symptoms such as
sneezing. Oral seed-based peptide immunotherapy offers a safe, simple, and
cost-effective alternative to conventional allergen-specific immunotherapy using
crude allergen extracts for treating allergic disease. A human version of rice
seed-based edible vaccine containing seven T-cell epitopes from the Cry j 1 and
Cry j 2 allergens was recently developed and is undergoing safety assessments.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17276883 [PubMed - indexed for MEDLINE]

807: Immunol Allergy Clin North Am. 2007 Feb;27(1):105-27.

New perspectives for use of native and engineered recombinant food proteins in
treatment of food allergy.

Nowak-Wegrzyn A.

Jaffe Food Allergy Institute, Division of Allergy and Immunology, Department of
Pediatrics, Mount Sinai School of Medicine, Box 1198, One G. Levy Place, NY
10029, USA. anna.nowak-wegrzyn@mssm.edu

Food allergy has emerged as an important target for research on curative
treatment and prevention, with most efforts focusing on peanut, cow's milk, and
egg allergy. This article reviews the recent developments in the potential
treatments for IgE-mediated food allergy using native and engineered recombinant
food proteins.

Publication Types:
Research Support, N.I.H., Extramural
Review

PMID: 17276882 [PubMed - indexed for MEDLINE]

808: Food Chem Toxicol. 2007 Apr;45(4):530-42. Epub 2006 Aug 25.

Approaches in the risk assessment of genetically modified foods by the Hellenic
Food Safety Authority.

Varzakas TH, Chryssochoidis G, Argyropoulos D.

Hellenic Food Safety Authority (EFET), Directorate of Nutritional Policy and
Research, Karystou 5, 115 23 Ampelokipoi, Greece. theovarzakas@yahoo.gr

Risk analysis has become important to assess conditions and take decisions on
control procedures. In this context it is considered a prerequisite in the
evaluation of GM food. Many consumers worldwide worry that food derived from
genetically modified organisms (GMOs) may be unhealthy and hence regulations on
GMO authorisations and labelling have become more stringent. Nowadays there is a
higher demand for non-GM products and these products could be differentiated from
GM products using the identity preservation system (IP) that could apply
throughout the grain processing system. IP is the creation of a transparent
communication system that encompasses HACCP, traceability and related systems in
the supply chain. This process guarantees that certain characteristics of the
lots of food (non-GM origin) are maintained "from farm to fork". This article
examines the steps taken by the Hellenic Food Safety Authority to examine the
presence of GMOs in foods. The whole integrated European legislation framework
currently in place still needs to be implemented in Greece. Penalties should be
enforced to those who import, process GMOs without special licence and do not
label those products. Similar penalties should be enforced to those companies
that issue false certificates beyond the liabilities taken by the food
enterprises for farmers' compensation. We argue that Greece has no serious
reasons to choose the use of GMOs due to the fact that the structural and
pedologic characteristics of the Greek agriculture favour the biological and
integrated cultivation more. Greece is not in favour of the politics behind
coexistence of conventional and GM plants and objects to the use of GMOs in the
food and the environment because the processor has a big burden in terms of
money, time and will suffer a great deal in order to prove that their products
are GMO free or that any contamination is adventitious or technically
unavoidable. Moreover, Greece owns a large variety of genetic material that
should try to protect from patenting and commercialisation. Finally, we should be
aware of the requirements of movement of GMOs within borders, i.e. GMOs grown or
used in other countries but which are not intended to cross into Greece, since
Greece is very close to countries that are non-EU. This is where the development
of a new, integrated, trustworthy and transparent food quality control system
will help to satisfy the societal demands for safe and quality products. On the
other hand, Greece should not be isolated from any recent scientific
technological development and should assess the possible advantages for some
cultivation using a case by case approach. Finally, the safety assessment of GM
foods and feed has been discussed according to the risk assessment methodology
applied by EFSA.

Publication Types:
Review

PMID: 17275157 [PubMed - indexed for MEDLINE]

809: Transgenic Res. 2007 Dec;16(6):771-81. Epub 2007 Feb 2.

pORE: a modular binary vector series suited for both monocot and dicot plant
transformation.

Coutu C, Brandle J, Brown D, Brown K, Miki B, Simmonds J, Hegedus DD.

Agriculture and Agrifood Canada, Saskatoon Research Centre, 107 Science Place,
Saskatoon, SK, Canada S7N 0X2.

We present a series of 14 binary vectors suitable for Agrobacterium-mediated
transformation of dicotyledonous plants and adaptable for biolistic
transformation of monocotyledonous plants. The vector size has been minimized by
eliminating all non-essential elements from the vector backbone and T-DNA regions
while maintaining the ability to replicate independently. The smallest of the
vector series is 6.3 kb and possesses an extensive multiple cloning site with 21
unique restriction endonuclease sites that are compatible with common cloning,
protein expression, yeast two-hybrid and other binary vectors. The T-DNA region
was engineered using a synthetic designer oligonucleotide resulting in an
entirely modular system whereby any vector element can be independently
exchanged. The high copy number ColE1 origin of replication has been included to
enhance plasmid yield in Escherichia coli. FRT recombination sites flank the
selectable marker cassette regions and allow for in planta excision by FLP
recombinase. The pORE series consists of three basic types; an 'open' set for
general plant transformation, a 'reporter' set for promoter analysis and an
'expression' set for constitutive expression of transgenes. The sets comprise
various combinations of promoters (P (HPL), P (ENTCUP2) and P (TAPADH)),
selectable markers (nptII and pat) and reporter genes (gusA and smgfp).

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17273915 [PubMed - indexed for MEDLINE]

810: Plant Mol Biol. 2007 Apr;63(6):847-60. Epub 2007 Feb 2.

Isolation and molecular characterization of a Spotted leaf 18 mutant by modified
activation-tagging in rice.

Mori M, Tomita C, Sugimoto K, Hasegawa M, Hayashi N, Dubouzet JG, Ochiai H,
Sekimoto H, Hirochika H, Kikuchi S.

National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.
morimasa@nias.affrc.go.jp

A lesion mimic mutant that we designated Spotted leaf 18 (Spl18) was isolated
from 13,000 activation-tagging lines of rice produced by our modified
activation-tagging vector and further characterized. Spl18 was dominant and its
phenotype was linked to the T-DNA insertion. An ORF was located about 500 bp
downstream of the inserted T-DNA, and the deduced protein, designated OsAT1,
showed sequence similarity to an acyltransferase whose expression is induced by
hypersensitive reaction in tobacco. The transcriptional level of OsAT1 was very
low in the WT leaf blade but high in Spl18 leaf blade. In wild-type rice, OsAT1
was transcribed mainly in the young panicle, in the panicle just after heading,
and in the leaf sheath. In addition, transcription of the genes for PR protein
was upregulated in Spl18, accumulation of phytoalexins (both momilactone A and
sakuranetin) was increased, and resistance to blast disease was improved. We then
combined OsAT1 genomic DNA downstream of the modified 35S promoter and
re-transformed it into rice. Lesion mimic and blast resistance phenotypes were
detected in the transgenic lines produced, clearly indicating that overexpression
of OsAT1 caused the Spl18 phenotypes. In addition, plants overexpressing OsAT1
showed resistance to bacterial blight.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17273822 [PubMed - indexed for MEDLINE]

811: Plant Mol Biol. 2007 May;64(1-2):89-101. Epub 2007 Feb 2.

Cladosporium fulvum CfHNNI1 induces hypersensitive necrosis, defence gene
expression and disease resistance in both host and nonhost plants.

Cai XZ, Zhou X, Xu YP, Joosten MH, de Wit PJ.

Institute of Biotechnology, and Department of Plant Protection, Zhejiang
University, 268 Kai Xuan Road, Hangzhou 310029, P.R. China. xzhcai@zju.edu.cn

Nonhost resistance as a durable and broad-spectrum defence strategy is of great
potential for agricultural applications. We have previously isolated a cDNA
showing homology with genes encoding bZIP transcription factors from tomato leaf
mould pathogen Cladosporium fulvum. Upon expression, the cDNA results in necrosis
in C. fulvum host tomato and nonhost tobacco plants and is thus named CfHNNI1
(for C . f ulvum host and nonhost plant necrosis inducer 1). In the present study
we report the induction of necrosis in a variety of nonhost plant species
belonging to three families by the transient in planta expression of CfHNNI1
using virus-based vectors. Additionally, transient expression of CfHNNI1 also
induced expression of the HR marker gene LeHSR203 and greatly reduced the
accumulation of recombinant Potato virus X. Stable CfHNNI1 transgenic tobacco
plants were generated in which the expression of CfHNNI1 is under the control of
the pathogen-inducible hsr203J promoter. When infected with the oomycetes
pathogen Phytophthora parasitica var. nicotianae, these transgenic plants
manifested enhanced expression of CfHNNI1 and subsequent accumulation of CfHNNI1
protein, resulting in high expression of the HSR203J and PR genes, and strong
resistance to the pathogen. The CfHNNI1 transgenic plants also exhibited induced
resistance to Pseudomonas syringae pv. tabaci and Tobacco mosaic virus.
Furthermore, CfHNNI1 was highly expressed and the protein was translocated into
plant cells during the incompatible interactions between C. fulvum and host and
nonhost plants. Our results demonstrate that CfHNNI1 is a potential general
elicitor of hypersensitive response and nonhost resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17273821 [PubMed - indexed for MEDLINE]

812: J Appl Genet. 2007;48(1):47-61.

Animal transgenesis: state of the art and applications.

Melo EO, Canavessi AM, Franco MM, Rumpf R.

EMBRAPA Genetic Resources and Biotechnology, Av. W/5, Norte Final, PBI, Sala 7B,
Brasilia, DF, Brazil, CEP 70770-900. eom@cenargen.embrapa.br

There is a constant expectation for fast improvement of livestock production and
human health care products. The advent of DNA recombinant technology and the
possibility of gene transfer between organisms of distinct species, or even
distinct phylogenic kingdoms, has opened a wide range of possibilities. Nowadays
we can produce human insulin in bacteria or human coagulation factors in cattle
milk. The recent advances in gene transfer, animal cloning, and assisted
reproductive techniques have partly fulfilled the expectation in the field of
livestock transgenesis. This paper reviews the recent advances and applications
of transgenesis in livestock and their derivative products. At first, the state
of art and the techniques that enhance the efficiency of livestock transgenesis
are presented. The consequent reduction in the cost and time necessary to reach a
final product has enabled the multiplication of transgenic prototypes around the
world. We also analyze here some emerging applications of livestock transgenesis
in the field of pharmacology, meat and dairy industry, xenotransplantation, and
human disease modeling. Finally, some bioethical and commercial concerns raised
by the transgenesis applications are discussed.

Publication Types:
Historical Article
Review

PMID: 17272861 [PubMed - indexed for MEDLINE]

813: Proc Natl Acad Sci U S A. 2007 Feb 6;104(6):1739-40. Epub 2007 Feb 1.

Comment on:
Proc Natl Acad Sci U S A. 2007 Feb 6;104(6):1771-6.

The incredible, edible, and therapeutic egg.

Petitte JN, Mozdziak PE.

Department of Poultry Science, College of Agriculture and Life Sciences, North
Carolina State University, Raleigh, NC 27695-7608, USA. j_petitte@ncsu.edu

Publication Types:
Comment

PMID: 17272493 [PubMed - indexed for MEDLINE]

814: Ann Bot (Lond). 2007 Mar;99(3):439-50. Epub 2007 Feb 1.

A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene
provides enhanced drought tolerance in tobacco.

Xu C, Jing R, Mao X, Jia X, Chang X.

The National Key Facility for Crop Gene Resources and Genetic Improvement, Key
Laboratory of Crop Germplasm & Biotechnology, Ministry of Agriculture, Institute
of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081,
China.

BACKGROUND AND AIMS: Multiple copies of genes encoding the catalytic subunit (c)
of protein phosphatase 2A (PP2A) are commonly found in plants. For some of these
genes, expression is up-regulated under water stress. The aim of this study was
to investigate expression and characterization of TaPP2Ac-1 from Triticum
aestivum, and to evaluate the effects of TaPP2Ac-1 on Nicotiana benthamiana in
response to water stress. METHODS: TaPP2Ac-1 cDNA was isolated from wheat by in
silico identification and RT-PCR amplification. Transcript levels of TaPP2Ac-1
were examined in wheat responding to water deficit. Copy numbers of TaPP2Ac-1 in
wheat genomes and subcellular localization in onion epidermal cells were studied.
Enzyme properties of the recombinant TaPP2Ac-1 protein were determined. In
addition, studies were carried out in tobacco plants with pCAPE2-TaPP2Ac-1 under
water-deficit conditions. KEY RESULTS: TaPP2Ac-1 cDNA was cloned from wheat.
Transcript levels of TaPP2Ac-1 in wheat seedlings were up-regulated under drought
condition. One copy for this TaPP2Ac-1 was present in each of the three wheat
genomes. TaPP2Ac-1 fused with GFP was located in the nucleus and cytoplasm of
onion epidermis cells. The recombinant TaPP2Ac-1 gene was over-expressed in
Escherichia coli and encoded a functional serine/threonine phosphatase.
Transgenic tobacco plants over-expressing TaPP2Ac-1 exhibited stronger drought
tolerance than non-transgenic tobacco plants. CONCLUSIONS: Tobacco plants with
pCAPE2-TaPP2Ac-1 appeared to be resistant to water deficit, as shown by their
higher capacity to maintain leaf relative water content, leaf cell-membrane
stability index, water-retention ability and water use efficiency under water
stress. The results suggest that the physiological role of TaPP2Ac-1 is related
to drought stress response, possibly through its involvement in
drought-responding signal transduction pathways.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17272305 [PubMed - indexed for MEDLINE]

815: Transgenic Res. 2007 Jun;16(3):353-61. Epub 2007 Jan 31.

Expression of recombinant human factor VIII in milk of several generations of
transgenic rabbits.

Chrenek P, Ryban L, Vetr H, Makarevich AV, Uhrin P, Paleyanda RK, Binder BR.

Slovak Agricultural Research Centre, Nitra, Slovak Republic. chrenekp@yahoo.com

Transgenic founder rabbits carrying a gene construct consisting of a 2.5 kb
murine whey acidic protein promoter (mWAP), 7.2 kb of the human clotting factor
VIII (hFVIII) cDNA and 4.6 kb of 3' flanking sequences of mWAP gene were crossed
for three generations. All transgenic animals showed stable transgene
transmission. Transgenic females showed high level of recombinant hFVIII
(rhFVIII) mRNA expression in biopsed mammary gland tissues, while marginal
expression of rhFVIII mRNA was observed in the spleen, lung and brain. No adverse
effects of ectopic expression on the physiology of the rabbits were observed.
Expression was not detected in the liver, kidney, heart and skeletal muscle. In
transgenic females derived from three generations, rhFVIII protein was secreted
from the mammary gland of lactating females, as shown by Western blotting.
Biological activity of rhFVIII protein, as revealed in clotting assays was ranged
from 0.012 to 0.599 IU/ml corresponding to 1.2% and 59.9% of the hFVIII level in
normal human plasma. No apparent effect of secreted rhFVIII on the milk
performance of rabbits was observed. Our results confirm the possibility of
producing a significant amount of a biologically active rhFVIII in the mammary
gland of established transgenic rabbit lines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17265165 [PubMed - indexed for MEDLINE]

816: Am J Physiol Endocrinol Metab. 2007 May;292(5):E1483-94. Epub 2007 Jan 30.

Adiposity profile in the dwarf rat: an unusually lean model of profound growth
hormone deficiency.

Davies JS, Gevers EF, Stevenson AE, Coschigano KT, El-Kasti MM, Bull MJ, Elford
C, Evans BA, Kopchick JJ, Wells T.

School of Biosciences, Cardiff University, Cardiff, UK.

This study describes the previously uncharacterized ontogeny and regulation of
truncal adipose reserves in the profoundly GH-deficient dwarf (dw/dw) rat. We
show that, despite normal proportionate food intake, dw/dw rats develop abdominal
leanness and hypoleptinemia (circulating leptin halved in dw/dw males, P < 0.05)
during puberty. This contrasts with the hyperleptinemia seen in moderately
GH-deficient Tgr rats (circulating leptin doubled at 6 wk of age, P < 0.05) and
in GH receptor-binding protein (GHR/BP)-null mice (circulating leptin doubled; P
< 0.05). This lean/hypoleptinemic phenotype was not completely normalized by GH
treatment, but dw/dw rats developed abdominal obesity in response to neonatal MSG
treatment or maintenance on a high-fat diet. Unlike Tgr rats, dw/dw rats did not
become obese with age; plasma leptin levels and fat pad weights became similar to
those in wild-type rats. In contrast with truncal leanness, tibial marrow
adiposity was normal in male and doubled in female dwarves (P < 0.01), this
increase being attributable to increased adipocyte number (P < 0.01). Neonatal
MSG treatment and high-fat feeding elevated marrow adiposity in dw/dw rats by
inducing adipocyte enlargement (P < 0.05). These results demonstrate that,
despite lipolytic influence of GH, severe GH deficiency in dw/dw rats is
accompanied by a paradoxical leanness. This lean/hypoleptinemic phenotype is not
solely attributable to reduced GH signaling and does not appear to result from a
reduction in nutrient intake or the ability of dw/dw adipocytes to accumulate
lipid. Disruption of preadipocyte differentiation or adipocyte proliferation in
the dw/dw rat may lead to the development of this unusually lean/hypoleptinemic
phenotype.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17264226 [PubMed - indexed for MEDLINE]

817: Proc Natl Acad Sci U S A. 2007 Feb 6;104(6):1766-70. Epub 2007 Jan 29.

Transgenic induction of mitochondrial rearrangements for cytoplasmic male
sterility in crop plants.

Sandhu AP, Abdelnoor RV, Mackenzie SA.

Plant Science Initiative, University of Nebraska, Lincoln, NE 68505, USA.

Stability of the mitochondrial genome is controlled by nuclear loci. In plants,
nuclear genes suppress mitochondrial DNA rearrangements during development. One
nuclear gene involved in this process is Msh1. Msh1 appears to be involved in the
suppression of illegitimate recombination in plant mitochondria. To test the
hypothesis that Msh1 disruption leads to the type of mitochondrial DNA
rearrangements associated with naturally occurring cytoplasmic male sterility in
plants, a transgenic approach for RNAi was used to modulate expression of Msh1 in
tobacco and tomato. In both species, these experiments resulted in reproducible
mitochondrial DNA rearrangements and a condition of male (pollen) sterility. The
male sterility was, in each case, heritable, associated with normal female
fertility, and apparently maternal in its inheritance. Segregation of the
transgene did not reverse the male sterile phenotype, producing stable,
nontransgenic male sterility. The reproducible transgenic induction of
mitochondrial rearrangements in plants is unprecedented, providing a means to
develop novel cytoplasmic male sterile lines for release as non-GMO or transgenic
materials.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 17261806 [PubMed - indexed for MEDLINE]

818: Proc Natl Acad Sci U S A. 2007 Feb 6;104(6):1771-6. Epub 2007 Jan 26.

Comment in:
Proc Natl Acad Sci U S A. 2007 Feb 6;104(6):1739-40.

Oviduct-specific expression of two therapeutic proteins in transgenic hens.

Lillico SG, Sherman A, McGrew MJ, Robertson CD, Smith J, Haslam C, Barnard P,
Radcliffe PA, Mitrophanous KA, Elliot EA, Sang HM.

Roslin Institute, Roslin Biocentre, Midlothian EH25 9PS, United Kingdom.

Recent advances in avian transgenesis have led to the possibility of utilizing
the laying hen as a production platform for the large-scale synthesis of
pharmaceutical proteins. Ovalbumin constitutes more than half of the protein in
the white of a laid egg, and expression of the ovalbumin gene is restricted to
the tubular gland cells of the oviduct. Here we describe the use of lentiviral
vectors to deliver transgene constructs comprising regulatory sequences from the
ovalbumin gene designed to direct synthesis of associated therapeutic proteins to
the oviduct. We report the generation of transgenic hens that synthesize
functional recombinant pharmaceutical protein in a tightly regulated
tissue-specific manner, without any evidence of transgene silencing after
germ-line transmission.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17259305 [PubMed - indexed for MEDLINE]

819: Phytochemistry. 2007 Mar;68(6):797-801. Epub 2007 Jan 26.

Reduction of divinyl ether-containing polyunsaturated fatty acids in transgenic
potato plants.

Eschen-Lippold L, Rothe G, Stumpe M, Gö¶bel C, Feussner I, Rosahl S.

Department of Stress and Developmental Biology, Leibniz Institute of Plant
Biochemistry, Weinberg 3, D-06120 Halle/Saale, Germany.

Oxygenated polyunsaturated fatty acids synthesized via the lipoxygenase pathway
play a role in plant responses to pathogen attack. In solanaceous plants, the
preferential stimulation of the 9-lipoxygenase pathway in response to pathogen
infection leads to the formation of the divinyl ether-containing polyunsaturated
fatty acids colneleic and colnelenic acid, as well as hydroxy and trihydroxy
polyunsaturated fatty acids. To functionally assess the role of divinyl ethers,
transgenic potato plants were generated which express an RNA interference
construct directed against the pathogen-inducible 9-divinyl ether synthase.
Efficient reduction of 9-divinyl ether synthase transcript accumulation
correlated with reduced levels of colneleic and colnelenic acid. However, in
response to infection with virulent Phytophthora infestans, the causal agent of
late blight disease, no significant differences in pathogen biomass could be
detected suggesting that the levels of antimicrobial divinyl ethers are not
critical for defense against Phytophthora infestans in a compatible interaction.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17258245 [PubMed - indexed for MEDLINE]

820: Plant J. 2007 Mar;49(5):910-23. Epub 2007 Jan 26.

The PEP-carboxylase kinase gene family in Glycine max (GmPpcK1-4): an in-depth
molecular analysis with nodulated, non-transgenic and transgenic plants.

Xu W, Sato SJ, Clemente TE, Chollet R.

Department of Biochemistry, University of Nebraska-Lincoln, George W. Beadle
Center, Lincoln, NE 68588-0664, USA.

Phosphoenolpyruvate carboxylase (PEPC) is a widely distributed metabolic enzyme
among plant and prokaryotic species. In vascular plants, the typical PEPC is
regulated post-translationally by a complex interplay between opposing metabolite
effectors and reversible protein phosphorylation. This phosphorylation event is
controlled primarily by the up-/down-regulation of PEPC-kinase (PpcK), an
approximately 31-kDa Ser/Thr-kinase. As a sequel to earlier investigations
related to PEPC phosphorylation in N(2)-fixing nodules of Glycine max, we now
present a detailed molecular analysis of the PpcK multigene family in nodulated
soybeans. Although the GmPpcK1-4 transcripts are all expressed throughout nodule
development, only the nearly identical GmPpcK2/3 homologs are nodule-enhanced and
up-/down-regulated in vivo by photosynthate supply from the shoots. In contrast,
GmPpcK1 is a 'housekeeping' gene, and GmPpcK4 is a highly divergent member,
distantly removed from the legume PpcK subfamily. Real-time qRT-PCR analysis
indicates that GmPpcK2/3 are overwhelmingly the dominant PpcKs expressed and
up-/down-regulated throughout nodule development, mirroring the expression
properties of nodule-enhanced PEPC (GmPpc7). In situ RT-PCR investigation of the
spatial localization of the GmPpcK1-4 and GmPpc7 transcripts in mature nodules is
entirely consistent with this view. Complementary histochemical and related RNA
gel-blot findings with nodulated, GmPpcK1/3 promoter::GUS-expressing T(2) plants
provide direct experimental evidence that (i) PpcK gene expression is controlled
primarily at the transcriptional level; and (ii) the contrasting expression
properties of GmPpcK1/3 are conferred largely by regulatory element(s) within the
approximately 1.4-kb 5'-upstream region. As a result of our multifaceted analyses
of GmPpcK1-4, GmPpc7 and PEPC-phosphorylation in the soybean nodule, it is
proposed that the GmPpcK2/3 homologs and GmPpc7 together comprise the key
molecular 'downstream players' in this regulatory phosphorylation system within
the mature nodule's central zone.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17257170 [PubMed - indexed for MEDLINE]

821: Mol Ecol. 2007 Feb;16(3):487-99.

Modelling and estimating pollen movement in oilseed rape (Brassica napus) at the
landscape scale using genetic markers.

Devaux C, Lavigne C, Austerlitz F, Klein EK.

Laboratoire Ecologie, Systö©matique et Evolution, UMR CNRS-UPS-ENGREF 8079,
Bö¢timent 360, Universitö© Paris-Sud, F 91405 Orsay cedex, France.
celine.devaux@ese.u-psud.fr

Understanding patterns of pollen movement at the landscape scale is important for
establishing management rules following the release of genetically modified (GM)
crops. We use here a mating model adapted to cultivated species to estimate
dispersal kernels from the genotypes of the progenies of male-sterile plants
positioned at different sampling sites within a 10 x 10-km oilseed rape
production area. Half of the pollen clouds sampled by the male-sterile plants
originated from uncharacterized pollen sources that could consist of both large
volunteer and feral populations, and fields within and outside the study area.
The geometric dispersal kernel was the most appropriate to predict pollen
movement in the study area. It predicted a much larger proportion of
long-distance pollination than previously fitted dispersal kernels. This
best-fitting mating model underestimated the level of differentiation among
pollen clouds but could predict its spatial structure. The estimation method was
validated on simulated genotypic data, and proved to provide good estimates of
both the shape of the dispersal kernel and the rate and composition of pollen
issued from uncharacterized pollen sources. The best dispersal kernel fitted
here, the geometric kernel, should now be integrated into models that aim at
predicting gene flow at the landscape level, in particular between GM and non-GM
crops.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17257108 [PubMed - indexed for MEDLINE]

822: J Agric Food Chem. 2007 Feb 21;55(4):1241-7. Epub 2007 Jan 25.

Chemically induced expression of rice OSB2 under the control of the OsPR1.1
promoter confers increased anthocyanin accumulation in transgenic rice.

Kawahigashi H, Hirose S, Iwai T, Ohashi Y, Sakamoto W, Maekawa M, Ohkawa Y.

National Institute of Agrobiological Sciences (NIAS), 2-1-2 Kannondai, Tsukuba,
Ibaraki 305-8602, Japan. shiwak@affrc.go.jp

Anthocyanin pigmentation provides an excellent system with which to study the
regulation of gene expression in higher plants. In this study, OsPR1.1 promoter
was isolated and the promoter activity was monitored using a reporter gene OSB2,
which encodes a transcription factor for anthocyanin synthesis in rice plants. We
introduced PR::OSB2 plasmid into an isogenic Taichung 65, no. 99-962 T-65 CBA
B9F5 (T65 CBA), rice line (Oryza sativa L.) and found that the transgenic rice
plants exhibited anthocyanin accumulation by the induced expression of OSB2 after
chemical treatments with methyl jasmonate (MeJA) and 2,6-dichloroisonicotinic
acid (DCINA). The shoots of the PR::OSB2 transgenic rice plants changed color to
red after application of the chemicals accompanying with the increased
anthocyanin content to approximately 5-fold by MeJA and 2-fold by DCINA,
respectively. The anthocyanin accumulation was consistent with the increase of
the expression of OSB2 and anthocyanidin synthase (ANS). This color change system
could provide a useful and easy way to produce transgenic plants for monitoring
of chemicals in the environment.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17253710 [PubMed - indexed for MEDLINE]

823: J Microbiol Methods. 2007 Apr;69(1):107-15. Epub 2006 Dec 19.

A double kill gene cassette for the positive selection of transforming
non-selective DNA segments in Acinetobacter baylyi BD413.

Harms K, de Vries J, Wackernagel W.

Genetics, Department of Biology and Environmental Sciences, Carl von Ossietzky
University Oldenburg, D-26111 Oldenburg, Germany.

Natural transformation has been widely used for the monitoring of DNA in
biological and environmental samples. These assays depended on selectable traits
on the tested DNA. We have now developed a transformation assay system in which
recombinational removal of a cassette with two conditional kill genes (hok and
sacB) from the recipient genome provides positive selection for non-selective
DNA. The cassette was integrated into the Acinetobacter baylyi BD413 chromosome
within trpE and was flanked by two segments of non-selective test DNA, which in
this study were from a T-DNA construct previously used to generate a transgenic
potato. Genes for tetracycline and spectinomycin/streptomycin resistance located
at the sides of the cassette allowed to maintain selection pressure against
spontaneous loss of the cassette. Plasmid DNA containing the T-DNA gave
transformation frequencies ranging linearly from 10(-4) per recipient (at 1 mug
DNA ml(-1)) down to 10(-7) (1 ng DNA ml(-1)) by selecting for survivors after
activation of both kill functions. Transformation depended on the two flanking
homologous segments for recombinational exchange. DNA of the transgenic potato
also gave positive scores in spite of the about 10(5)-fold dilution of T-DNA by
potato DNA. False positives having a spontaneous deletion of hok and sacB
occurred at a frequency of 1.8x10(-9) per cell but could be distinguished by PCR
from real transformants. Thus, the system is suitable for detection of
transformation frequencies down to about 10(-9). Hok and sacB as well as the
regulatory system used (LacI-lac operator and T5 promoter) are known to function
in many organisms suggesting wide applicability of the cassette for positive
selection.

PMID: 17250911 [PubMed - indexed for MEDLINE]

824: Mol Plant Microbe Interact. 2007 Jan;20(1):72-81.

The LeATL6-associated ubiquitin/proteasome system may contribute to fungal
elicitor-activated defense response via the jasmonic acid-dependent signaling
pathway in tomato.

Hondo D, Hase S, Kanayama Y, Yoshikawa N, Takenaka S, Takahashi H.

Department of Life Science, Graduate School of Agricultural Science, Tohoku
University, Sendai 981-8555, Japan.

The expression of LeATL6, an ortholog of Arabidopsis ATL6 that encodes a RING-H2
finger protein, was induced in tomato roots treated with a cell wall protein
fraction (CWP) elicitor of the biocontrol agent Pythium oligandrum. The LeATL6
protein was expressed as a fusion protein with a maltose-binding protein (MBP) in
Escherichia coli, and it catalyzed the transfer of ubiquitin to the MBP moiety on
incubation with ubiquitin, the ubiquitin-activating enzyme E1, and the
ubiquitin-conjugating enzyme E2; this indicated that LeATL6 represents ubiquitin
ligase E3. LeATL6 expression also was induced by elicitor treatment of jail-1
mutant tomato cells in which the jasmonic acid (JA)-mediated signaling pathway
was impaired; however, JA-dependent expression of the basic PR-6 and TPI-1 genes
that encode proteinase inhibitor II and I, respectively, was not induced in
elicitor-treated jail-1 mutants. Furthermore, transient overexpression of LeATL6
under the control of the Cauliflower mosaic virus 35S promoter induced the basic
PR6 and TPI-1 expression in wild tomato but not in the jail-1 mutant. In
contrast, LeATL6 overexpression did not activate salicylic acid-responsive acidic
PR-1 and PR-2 promoters in wild tomato. These results indicated that
elicitor-responsive LeATL6 probably regulates JA-dependent basic PR6 and TPI-1
gene expression in tomato. The LeATL6-associated ubiquitin/proteasome system may
contribute to elicitor-activated defense responses via a JA-dependent signaling
pathway in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17249424 [PubMed - indexed for MEDLINE]

825: Plant Mol Biol. 2007 May;64(1-2):49-58. Epub 2007 Jan 24.

Expression of an NADP-malic enzyme gene in rice (Oryza sativa. L) is induced by
environmental stresses; over-expression of the gene in Arabidopsis confers salt
and osmotic stress tolerance.

Liu S, Cheng Y, Zhang X, Guan Q, Nishiuchi S, Hase K, Takano T.

Alkali Soil Natural Environmental Science Center (ASNESC), Stress Molecular
Biology Laboratory, Northeast Forestry University, Harbin 150040, P. R. China.

NADP-malic enzyme (NADP-ME, EC 1.1.1.40) functions in many different pathways in
plants, and has recently been implicated in plant defense such as in responses to
wounding and UV-B radiation. In this study, we isolated a complementary DNA
(cDNA) clone by using the differential display method and screening of a root
cDNA library of rice (Oryza sativa. L) under carbonate (NaHCO3) stress, and
identified it as one of the rice NADP-ME genes (we named it NADP-ME2, GenBank
accession no. AB053295). The 5' end of NADP-ME2 was obtained by the 5'-RACE
method, and the full-length cDNA had a length of 2217 bp encoding 593 amino
acids. Expression of NADP-ME2 mRNA in roots was induced by stress from carbonates
(NaHCO3 and Na2CO3, NaCl, and environmental pH changes. NADP-ME2 transcripts
increased during 72-h exposures to NaHCO3, NaCl, and PEG stresses. Furthermore,
NADP-ME activities in leaves and roots of rice seedlings increased by more than
50% in the presence of carbonates (NaHCO3 and Na2CO3), NaCl, and PEG. These
results indicate that rice NADP-ME2 responds to salts and osmotic stresses.
Transgenic Arabidopsis plants over-expressing NADP-ME2 were obtained through
transformation, screening, Northern analysis and in situ NADP-ME activity assay.
Transgenic Arabidopsis plants over-expressing NADP-ME2 grew well in 1/2 x MS
medium with 100 mM NaCl or 4% mannitol, whereas growth of wild-type (WT)
Arabidopsis seedlings was strongly inhibited. In addition, under 125 mM NaCl
stress, the root lengths of transgenic lines were about twice as long as those of
the WT. These results suggest that NADP-ME2 has a role in enhancing tolerance of
plants to salt and osmotic stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17245561 [PubMed - indexed for MEDLINE]

826: J Agric Food Chem. 2007 Feb 21;55(4):1274-9. Epub 2007 Jan 23.

Development and evaluation of event-specific qualitative PCR methods for
genetically modified Bt10 maize.

Watanabe T, Tokishita S, Spiegelhalter F, Furui S, Kitta K, Hino A, Matsuda R,
Akiyama H, Maitani T.

National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo
158-8501, Japan. tawata@nihs.go.jp

In 2005 it was reported that the genetically modified (GM) maize strain or
"event" called Bt10 had been distributed inadvertently in the United States over
the previous 4 years. In order to ensure that grain for food and feed production
did not contain trace amounts of Bt10 maize and complied with the applicable
regulation, highly sensitive and specific detection of Bt10 maize was required.
Accordingly, we developed a novel qualitative PCR system for specific detection
of Bt10 maize. Moreover, we amply evaluated the performance characteristics of
two PCR systems, our own and the one provided by the developer of Bt10, Syngenta
Co. Ltd. It was confirmed that both of the qualitative PCR systems can
specifically detect Bt10 maize, and the results of a single-laboratory
examination suggested that the limit of detection was approximately less than
0.05% for both methods. To evaluate the reproducibility of the methods, we
organized an interlaboratory study with the participation of 6 laboratories and
analysis of 240 blind test samples. In this paper, we report, for the first time,
the statistical analysis of the qualitative PCR data obtained from the
interlaboratory study. The results of this analysis also revealed that there was
no significant difference in the sensitivity between the two aforementioned
methods and that the limit of detection of both the methods was less than 0.05%.
Thus, we conclude that both of the methods are equally suitable for correct
identification and sensitive detection of the unapproved GM maize Bt10 event in
test samples.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17243705 [PubMed - indexed for MEDLINE]

827: Plant J. 2007 Jan;49(2):276-88.

Contained metabolic engineering in tomatoes by expression of carotenoid
biosynthesis genes from the plastid genome.

Wurbs D, Ruf S, Bock R.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie (MPI-MP), Am Mö¼hlenberg 1,
D-14476 Potsdam-Golm, Germany.

Applications of chloroplast engineering in agriculture and biotechnology will
depend critically on success in extending the crop range of chloroplast
transformation, and on the feasibility of expressing transgenes in edible organs
(such as tubers and fruits), which often are not green and thus are much less
active in chloroplast gene expression. We have improved a recently developed
chloroplast-transformation system for tomato plants and applied it to engineering
one of the central metabolic pathways in fruits: carotenoid biosynthesis. We
report that plastid expression of a bacterial lycopene beta-cyclase gene results
in herbicide resistance and triggers conversion of lycopene, the main storage
carotenoid of tomatoes, to beta-carotene, resulting in fourfold enhanced
pro-vitamin A content of the fruits. Our results demonstrate the feasibility of
engineering nutritionally important biochemical pathways in non-green plastids by
transformation of the chloroplast genome.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17241450 [PubMed - indexed for MEDLINE]

828: Virol J. 2007 Jan 19;4:10.

Silencing of the AV2 gene by antisense RNA protects transgenic plants against a
bipartite begomovirus.

Mubin M, Mansoor S, Hussain M, Zafar Y.

Plant Biotechnology Division, National Institute for Biotechnology and Genetic
Engineering (NIBGE), P O Box 577, Jhang Road, Faisalabad, Pakistan.
mubin447@yahoo.com

Whitefly-transmitted geminiviruses (genus Begomovirus) are phytopathogens that
cause heavy losses to crops worldwide. Efforts to engineer resistance against
these viruses are focused mainly on silencing of complementary-sense virus genes
involved in virus replication. Here we have targeted a virion-sense gene (AV2) to
develop resistance against Tomato leaf curl New Delhi virus, a bipartite
begomovirus prevalent throughout the Indian subcontinent. We show that tobacco
plants transformed with an antisense construct targeting this gene are resistant
to the virus. Following challenged with the virus, transgenic plants remained
symptomless, although viral DNA could be detected in some plants by PCR. This is
the first report of transgenic resistance against a bipartite begomovirus
obtained by targeting a virion-sense gene. The relatively conserved nature of the
gene suggests that the technology may be useful to develop broad-spectrum
resistance which is required because of the fact that plants are often infected
with multiple begomoviruses in the field.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17239233 [PubMed - indexed for MEDLINE]

829: Transgenic Res. 2007 Dec;16(6):739-49. Epub 2007 Jan 20.

Enhanced conversion of plant biomass into glucose using transgenic rice-produced
endoglucanase for cellulosic ethanol.

Oraby H, Venkatesh B, Dale B, Ahmad R, Ransom C, Oehmke J, Sticklen M.

Department of Crop and Soil Sciences, Michigan State University, East Lansing, MI
48824, USA.

The catalytic domain of Acidothermus cellulolyticus thermostable endoglucanase
gene (encoding for endo-1,4-beta-glucanase enzyme or E1) was constitutively
expressed in rice. Molecular analyses of T1 plants confirmed presence and
expression of the transgene. The amount of E1 enzyme accounted for up to 4.9% of
the plant total soluble proteins, and its accumulation had no apparent
deleterious effects on plant growth and development. Approximately 22 and 30% of
the cellulose of the Ammonia Fiber Explosion (AFEX)-pretreated rice and maize
biomass respectively was converted into glucose using rice E1 heterologous
enzyme. As rice is the major food crop of the world with minimal use for its
straw, our results suggest a successful strategy for producing biologically
active hydrolysis enzymes in rice to help generate alcohol fuel, by substituting
the wasteful and polluting practice of rice straw burning with an environmentally
friendly technology.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17237981 [PubMed - indexed for MEDLINE]

830: J Exp Bot. 2007;58(5):947-56. Epub 2007 Jan 19.

Isolation and functional characterization of PgTIP1, a hormone-autotrophic
cells-specific tonoplast aquaporin in ginseng.

Lin W, Peng Y, Li G, Arora R, Tang Z, Su W, Cai W.

Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological
Sciences, Chinese Academy of Sciences, and Graduate School of the Chinese Academy
of Sciences, 300 Fenglin Road, Shanghai 200032, China.

The suppression subtractive hybridization technique was used to identify
differentially expressed genes between hormone-autotrophic and hormone-dependent
Panax ginseng callus lines. A tonoplast intrinsic protein cDNA (PgTIP1) was found
to be highly and specifically expressed in hormone-autotrophic ginseng cells,
which was slightly up-regulated by cytokinin while significantly down-regulated
when treated with auxin. PgTIP1 encodes a polypeptide of 250 amino acids which
shows sequence and structure similarity with tonoplast aquaporins in plants. The
water channel activity of PgTIP1 was demonstrated by its expression in Xenopus
laevis oocytes. When over-expressed in Arabidopsis thaliana, PgTIP1 substantially
altered the plant's vegetative and reproductive growth and development.
Arabidopsis plants over-expressing PgTIP1 showed significantly enhanced seed size
and seed mass plus greatly increased growth rate compared with those of the wild
type. Moreover, the seeds from PgTIP1 over-expressing Arabidopsis had 1.85-fold
higher fatty acid content than the wild-type control. These results demonstrate a
significant function of PgTIP1 in the growth and development of plant cells.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17237160 [PubMed - indexed for MEDLINE]

831: Plant Biol (Stuttg). 2007 May;9(3):447-52. Epub 2007 Jan 19.

A mutation in the AtPRP4 splicing factor gene suppresses seed development in
Arabidopsis.

Raab S, Hoth S.

Molekulare Pflanzenphysiologie, Friedrich-Alexander-Universitö¤t
Erlangen-Nö¼rnberg, Staudtstrasse 5, 91058 Erlangen, Germany.

The spliceosome catalyzes alternative splicing of many genes in eucaryotic cells.
This leads to the expression of distinct proteins. Components of the spliceosome
are conserved in mammals and plants. Because splicing can be affected by
environmental stress, we analyzed the regulation of splicing-related genes that
encode small nuclear ribonucleoprotein particle (snRNP) proteins by the stress
hormone abscisic acid (ABA). The transcript abundance of about 25 % of those
genes was changed by at least 1.5-fold after addition of ABA. The U4/U6-specific
snRNP gene AtPRP4 was strongly repressed by ABA. The homozygous knock-out of
AtPRP4 resulted in the suppression of seed development suggesting that the gene
product of this stress hormone-regulated gene is crucial for normal seed
development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17236097 [PubMed - indexed for MEDLINE]

832: Science. 2007 Jan 19;315(5810):381-4.

Comment in:
Science. 2007 Jan 19;315(5810):341-2.

Farmland biodiversity and the footprint of agriculture.

Butler SJ, Vickery JA, Norris K.

Centre for Agri-Environment Research, School of Agriculture, Policy, and
Development, University of Reading, Reading RG6 6AR, UK. s.j.butler@reading.ac.uk

Sustainable development requires the reconciliation of demands for biodiversity
conservation and increased agricultural production. Assessing the impact of novel
farming practices on biodiversity and ecosystem services is fundamental to this
process. Using farmland birds as a model system, we present a generic risk
assessment framework that accurately predicts each species' current conservation
status and population growth rate associated with past changes in agriculture. We
demonstrate its value by assessing the potential impact on biodiversity of two
controversial land uses, genetically modified herbicide-tolerant crops and
agri-environment schemes. This framework can be used to guide policy and land
management decisions and to assess progress toward sustainability targets.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17234947 [PubMed - indexed for MEDLINE]

833: Science. 2007 Jan 19;315(5810):341-2.

Comment on:
Science. 2007 Jan 19;315(5810):381-4.

Ecology. Managing farming's footprint on biodiversity.

Benton TG.

Institute of Integrative and Comparative Biology, University of Leeds, Leeds LS2
9JT, UK. t.g.benton@leeds.ac.uk

Publication Types:
Comment

PMID: 17234937 [PubMed - indexed for MEDLINE]

834: Biochem Biophys Res Commun. 2007 Mar 9;354(2):440-6. Epub 2007 Jan 9.

Abiotic and biotic stress tolerance in Arabidopsis overexpressing the
multiprotein bridging factor 1a (MBF1a) transcriptional coactivator gene.

Kim MJ, Lim GH, Kim ES, Ko CB, Yang KY, Jeong JA, Lee MC, Kim CS.

Department of Plant Biotechnology and Agricultural Plant Stress Research Center,
Chonnam National University, Kwangju 500-757, Republic of Korea.

We conducted a genetic yeast screen to identify salt tolerance (SAT) genes in a
maize kernel cDNA library. During the screening, we identified a maize clone
(SAT41) that seemed to confer elevated salt tolerance in comparison to control
cells. SAT41 cDNA encodes a 16-kDa protein which is 82.4% identical to the
Arabidopsis Multiprotein bridging factor 1a (MBF1a) transcriptional coactivator
gene. To further examine salinity tolerance in Arabidopsis, we functionally
characterized the MBF1a gene and found that dehydration as well as heightened
glucose (Glc) induced MBF1a expression. Constitutive expression of MBF1a in
Arabidopsis led to elevated salt tolerance in transgenic lines. Interestingly,
plants overexpressing MBF1a exhibited insensitivity to Glc and resistance to
fungal disease. Our results suggest that MBF1a is involved in stress tolerance as
well as in ethylene and Glc signaling in Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17234157 [PubMed - indexed for MEDLINE]

835: Lab Anim. 2007 Jan;41(1):30-45.

Should laboratory mice be anaesthetized for tail biopsy?

Arras M, Rettich A, Seifert B, Kö¤sermann HP, Rö¼licke T.

Institute of Laboratory Animal Science, University of Zurich, Sternwartstrasse 6,
CH-8091 Zurich, Switzerland. marras@bzl.unizh.ch

Tail biopsies are routinely taken to genotype genetically modified mice. However,
the effect of this procedure on the wellbeing of the animals has rarely been
investigated. Thus, it has not yet been clearly demonstrated to what extent the
mice suffer from tail biopsy (TB) and for how long. The aim of our study was to
assess the impact of a single TB on the physiological and behavioural parameters
of adult mice and to investigate whether or not anaesthesia can be beneficial.
Body weight (BW) curves, daily food/water consumption and telemetric measurements
of heart rate, body core temperature, and locomotor activity were recorded for
three days following TB, both with and without anaesthesia with methoxyflurane
(MOF) or diethylether (ether). Additionally, the impact of anaesthesia alone was
characterized. TB without anaesthesia induced an increase in heart rate and
locomotor activity for 1 h. Body core temperature was elevated for 2 h. In
contrast, heart rate was increased for up to 4 h after anaesthesia. Body core
temperature remained altered for up to 20 h after exposure to ether and for 44 h
after exposure to MOF. BW was slightly reduced after MOF. Cases of death occurred
exclusively under ether at a rate of 7%. Our results indicate a short-lived
impact of a TB, whereas anaesthesia with either MOF or ether induced remarkable
alterations in the parameters analysed. In conclusion, these types of anaesthesia
did not improve mouse wellbeing following tail biopsy.

PMID: 17234048 [PubMed - indexed for MEDLINE]

836: J Exp Bot. 2007;58(2):301-8. Epub 2007 Jan 17.

Overexpression of wheat Na+/H+ antiporter TNHX1 and H+-pyrophosphatase TVP1
improve salt- and drought-stress tolerance in Arabidopsis thaliana plants.

Brini F, Hanin M, Mezghani I, Berkowitz GA, Masmoudi K.

Plant Molecular Genetics Unit, Centre of Biotechnology of Sfax (CBS), B.P'K',
3038 Sfax, Tunisia.

Transgenic Arabidopsis plants overexpressing the wheat vacuolar Na(+)/H(+)
antiporter TNHX1 and H(+)-PPase TVP1 are much more resistant to high
concentrations of NaCl and to water deprivation than the wild-type strains. These
transgenic plants grow well in the presence of 200 mM NaCl and also under a
water-deprivation regime, while wild-type plants exhibit chlorosis and growth
inhibition. Leaf area decreased much more in wild-type than in transgenic plants
subjected to salt or drought stress. The leaf water potential was less negative
for wild-type than for transgenic plants. This could be due to an enhanced
osmotic adjustment in the transgenic plants. Moreover, these transgenic plants
accumulate more Na(+) and K(+) in their leaf tissue than the wild-type plants.
The toxic effect of Na(+) accumulation in the cytosol is reduced by its
sequestration into the vacuole. The rate of water loss under drought or salt
stress was higher in wild-type than transgenic plants. Increased vacuolar solute
accumulation and water retention could confer the phenotype of salt and drought
tolerance of the transgenic plants. Overexpression of the isolated genes from
wheat in Arabidopsis thaliana plants is worthwhile to elucidate the contribution
of these proteins to the tolerance mechanism to salt and drought. Adopting a
similar strategy could be one way of developing transgenic staple crops with
improved tolerance to these important abiotic stresses.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17229760 [PubMed - indexed for MEDLINE]

837: Plant J. 2006 Dec;48(6):920-32.

A mutation in Arabidopsis cytochrome b5 reductase identified by high-throughput
screening differentially affects hydroxylation and desaturation.

Kumar R, Wallis JG, Skidmore C, Browse J.

Institute of Biological Chemistry, Washington State University, Pullman, WA
99164-6340, USA.

As a model for analyzing the production of novel fatty acids in oilseeds, we used
the genetic and molecular techniques available for Arabidopsis to characterize
modifying mutations affecting the accumulation of hydroxy fatty acids in the
seeds of Arabidopsis plants that express a transgene for the castor bean fatty
acid hydroxylase, FAH12. We developed a high-throughput analytical system and
used it to identify three complementation classes of mutations with reduced
hydroxy fatty acid accumulation from among Arabidopsis M3 seed samples derived
from chemical mutagenesis. We identified one of the mutations by positional
cloning as a single base pair change in a gene encoding NADH:cytochrome b5
reductase (CBR1, At5g17770). When expressed in yeast, the mutant form of the
enzyme was less active and less stable than the wild-type enzyme.
Characterization of homozygous mutant lines with and without the FAH12 transgene
(FAH12 cbr1-1 and cbr1-1, respectively) indicated that the only detectable
consequence of the cbr1-1 mutation was on desaturase and hydroxylase reactions in
the developing seed. The leaf and root fatty compositions, as well as the growth,
development and seed production of mutant plants were indistinguishable from wild
type. Interestingly, while the cbr1-1 mutation reduced the accumulation of
hydroxy fatty acids in seeds by 85%, the effects on 18:1 and 18:2 desaturation
reactions were much less (<25% and <60%, respectively). These results suggest
that there is competition in developing seeds among the several reactions that
utilize reduced cytochrome b5.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17227547 [PubMed - indexed for MEDLINE]

838: J AOAC Int. 2006 Nov-Dec;89(6):1700-1.

Committee on microbiology and extraneous materials.

Agin JR, Ziemer WA, Newman MC, Guilfoyle DE, Vought K, Ledenbach L, Brodsky MH,
Hill W, Rice D, Ferreira JL, Werner BG, Martin BM, Shively R, Marrow T, Phillips
RW, Wehling P, Labudde RA; AOAC. Committee on microbiology and extraneous
materials.

Ohio Department of Agriculture, 8995 E. Main St, Reynoldsburg, OH 43068, USA.

Publication Types:
Guideline

PMID: 17225618 [PubMed - indexed for MEDLINE]

839: J Comp Physiol [B]. 2007 May;177(4):413-22. Epub 2007 Jan 16.

The glutathione antioxidant system is enhanced in growth hormone transgenic coho
salmon (Oncorhynchus kisutch).

Leggatt RA, Brauner CJ, Iwama GK, Devlin RH.

Faculty of Land and Food Systems, University of British Columbia, 2357 Main Mall,
Vancouver, BC, V6T 1Z4 Canada.

Insertion of a growth hormone (GH) transgene in coho salmon results in
accelerated growth, and increased feeding and metabolic rates. Whether other
physiological systems within the fish are adjusted to this accelerated growth has
not been well explored. We examined the effects of a GH transgene and feeding
level on the antioxidant glutathione and its associated enzymes in various
tissues of coho salmon. When transgenic and control salmon were fed to satiation,
transgenic fish had increased tissue glutathione, increased hepatic glutathione
reductase activity, decreased hepatic activity of the glutathione synthesis
enzyme gamma-glutamylcysteine synthetase, and increased intestinal activity of
the glutathione catabolic enzyme gamma-glutamyltranspeptidase. However, these
differences were mostly abolished by ration restriction and fasting, indicating
that upregulation of the glutathione antioxidant system was due to accelerated
growth, and not to intrinsic effects of the transgene. Increased food intake and
ability to digest potential dietary glutathione, and not increased activity of
glutathione synthesis enzymes, likely contributed to the higher levels of
glutathione in transgenic fish. Components of the glutathione antioxidant system
are likely upregulated to combat potentially higher reactive oxygen species
production from increased metabolic rates in GH transgenic salmon.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17225138 [PubMed - indexed for MEDLINE]

840: Appl Microbiol Biotechnol. 2007 May;75(1):55-60. Epub 2007 Jan 16.

Ergosterol production from molasses by genetically modified Saccharomyces
cerevisiae.

He X, Guo X, Liu N, Zhang B.

Laboratory of Yeast Molecular Genetics and Breeding, Institute of Microbiology,
Chinese Academy of Sciences, Beijing, PR China.

Ergosterol is an economically important metabolite produced by fungi. Recombinant
Saccharomyces cerevisiae YEH56(pHXA42) with increased capacity of ergosterol
formation was constructed by combined overexpression of sterol C-24(28) reductase
and sterol acyltransferase in the yeast strain YEH56. The production of
ergosterol by this recombinant strain using cane molasses (CM) as an inexpensive
carbon source was investigated. An ergosterol content of 52.6 mg/g was obtained
with 6.1 g/l of biomass from CM medium containing 60 g/l of total sugar in 30 h
in shake flask. The ergosterol yield was enhanced through the increasing cell
biomass by supplementation of urea to a concentration of 6 g/l in molasses
medium. Fermentation was performed in 5-l bioreactor using the optimized molasses
medium. In batch fermentation, the effect of agitation velocity on ergosterol
production was examined. The highest ergosterol yield was obtained at 400 rpm
that increased 60.4 mg/l in comparison with the shake flask culture. In fed-batch
fermentation, yeast cells were cultivated, firstly, in the starting medium
containing molasses with 20 g/l of total sugar, 1.68 g/l of phosphate acid, and 6
g/l of urea (pH 5.4) for 5 h, then molasses containing 350 g/l of total sugar was
fed exponentially into the bioreactor to keep the ethanol level in the broth
below 0.5%. After 40 h of cultivation, the ergosterol yield reached 1,707 mg/l,
which was 3.1-fold of that in the batch fermentation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17225097 [PubMed - indexed for MEDLINE]

841: Transgenic Res. 2007 Apr;16(2):169-75. Epub 2007 Jan 16.

Cholera toxin B protein in transgenic tomato fruit induces systemic immune
response in mice.

Jiang XL, He ZM, Peng ZQ, Qi Y, Chen Q, Yu SY.

School of Public Health and Tropical Medicine, Southern Medical University,
Guangzhou, 510515, P.R. China.

Cholera toxin B (CTB) subunit is a well-characterized antigen against cholera.
Transgenic plants can offer an inexpensive and safe source of edible CTB vaccine
and may be one of the best candidates for the production of plant vaccines. The
present study aimed to develop transgenic tomato expressing CTB protein,
especially in the ripening tomato fruit under the control of the tomato
fruit-specific E8 promoter by using Agrobacterium-mediated transformation.
Transgenic plants were selected using PCR and Southern blot analysis. Exogenous
protein extracted from leaf, stem, and fruit tissues of transgenic plants was
detected by ELISA and Western blot analysis, showing specific expression in the
ripening fruit, with the highest amount of CTB protein being 0.081% of total
soluble protein. Gavage of mice with ripe transgenic tomato fruits induced both
serum and mucosal CTB specific antibodies. These results demonstrate the
immunogenicity of the CTB protein in transgenic tomato and provide a considerable
basis for exploring the utilization of CTB in the development of tomato-based
edible vaccine against cholera. The rCTB antigen resulted in much lower antibody
titers than an equal amount of exogenous CTB in transgenic fruits, suggesting the
protective effect of the fibrous tissue of the fruit to the exogenous CTB protein
against the degradation of protease in the digestive tracts of mice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17225072 [PubMed - indexed for MEDLINE]

842: J Plant Physiol. 2007 Jul;164(7):824-34. Epub 2007 Jan 16.

Specificity of expression of the GUS reporter gene (uidA) driven by the tobacco
ASA2 promoter in soybean plants and tissue cultures.

Inaba Y, Zhong WQ, Zhang XH, Widholm JM.

Department of Crop Sciences, Edward R. Madigan Laboratory, University of
Illinois, 1201 W. Gregory Drive, Urbana, IL 61801, USA.

Twelve independent lines were transformed by particle bombardment of soybean
embryogenic suspension cultures with the tobacco anthranilate synthase (ASA2)
promoter driving the uidA (beta-glucuronidase, GUS) reporter gene. ASA2 appears
to be expressed in a tissue culture specific manner in tobacco (Song H-S,
Brotherton JE, Gonzales RA, Widholm JM. Tissue culture specific expression of a
naturally occurring tobacco feedback-insensitive anthranilate synthase. Plant
Physiol 1998;117:533-43). The transgenic lines also contained the hygromycin
phosphotransferase (hpt) gene and were selected using hygromycin. All the
selected cultures or the embryos that were induced from these cultures expressed
GUS measured histochemically. However, no histochemical GUS expression could be
found in leaves, stems, roots, pods and root nodules of the plants formed from
the embryos and their progeny. Pollen from some of the plants and immature and
mature seeds and embryogenic cultures initiated from immature cotyledons did show
GUS activity. Quantitative 4-methylumbelliferyl-glucuronide (MUG) assays of the
GUS activity in various tissues showed that all with observable histochemical GUS
activity contained easily measurable activities and leaves and stems that showed
no observable histochemical GUS staining did contain very low but measurable MUG
activity above that of the untransformed control but orders of magnitude lower
than the constitutive 35S-uidA controls used. Low but clearly above background
levels of boiling sensitive GUS activity could be observed in the untransformed
control immature seeds and embryogenic cultures using the MUG assay. Thus in
soybean the ASA2 promoter drives readily observable GUS expression in tissue
cultures, pollen and seeds, with only extremely low levels seen in vegetative
tissues of the plants. The ASA2 driven expression seen in mature seed was,
however, much lower than that seen with the constitutive 35S promoter; less than
2% in seed coats and less than 0.13% in cotyledons and embryo axes. The
predominate tissue culture specific expression pattern of the ASA2 promoter may
be useful for genetic transformation of crops.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17223226 [PubMed - indexed for MEDLINE]

843: J Chromatogr A. 2007 Feb 23;1142(2):137-47. Epub 2006 Dec 17.

The influence of major components on the direct chromatographic recovery of a
protein from transgenic milk.

Pampel L, Boushaba R, Udell M, Turner M, Titchener-Hooker N.

The Advanced Centre for Biochemical Engineering, University College London,
Torrington Place, London WC1E 7JE, UK.

This work presents a systematic evaluation of the influence of lipids and casein
on the performance of a chromatographic capture step for the recovery of a target
protein from transgenic milk. Lactoperoxidase (LPO) was spiked at concentrations
typical of those to be expected for transgenic proteins in commercial bovine milk
and the dynamic adsorption of LPO to fixed beds of SP Sepharose FF studied in
frontal analysis experiments. By removing successively selected components from
whole milk, their individual influence on the dynamic adsorption behaviour of LPO
could be studied. A mathematical model, fitted to the breakthrough curves of LPO,
provided a quantitative measure of parameters describing mass transfer and
adsorption in the column. A significant reduction in column capacity for LPO in
the presence of milk or whey was recorded, which could be attributed to competing
adsorption of alkaline earth metal ions to the cation exchange resin. While the
high concentrations of lipids present in whole milk did strongly reduce the
column permeability, no significant influence of either casein or low
concentrations of lipids on the hydraulic properties of columns or on the
adsorption of LPO could be detected. The results indicate that chromatography,
which forms an essential part of all current large-scale processes for the
recovery of proteins from transgenic milk, could potentially be moved further
upstream. Alternatively, existing operations for the removal of lipid and casein
could be re-designed so as to maximise product yields. This suggests that
significant product losses during current pre-chromatography milk purification
could be reduced or potentially even avoided.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17222855 [PubMed - indexed for MEDLINE]

844: Planta. 2007 Jun;226(1):73-85. Epub 2007 Jan 13.

Characterization of a stress responsive proteinase inhibitor gene with positive
effect in improving drought resistance in rice.

Huang Y, Xiao B, Xiong L.

National Center of Plant Gene Research (Wuhan), National Key Laboratory of Crop
Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China.

A full-length cDNA gene, designated Oryza sativa chymotrypsin inhibitor-like 1
(OCPI1), was characterized in rice. The predicted protein of OCPI1 shows very
high sequence identity to reported chymotrypsin inhibitors from various plant
species. Northern-blot analysis showed that the expression of OCPI1 was strongly
induced by dehydration stresses and abscisic acid (ABA). The expression of
beta-glucuronidase (GUS) reporter gene under the control of OCPI1 promoter
transformed into rice was strongly induced by drought and salt stresses.
Interestingly, strong dehydration stress-induced GUS activity was also detected
in the transgenic rice containing the reverse sequence of OCPI1 promoter fused to
GUS gene, suggesting of a bidirectional transcriptional activity in the OCPI1
promoter. OCPI1 gene was over-expressed in japonica cv. Zhonghua 11 and
transgenic plants containing single copy of transgene were tested for drought
resistance at reproductive stage. The positive transgenic plants (OCPI1 was
over-expressed) had significantly higher grain yield and seed setting rate than
the wild type and the negative transgenic control (no over-expression of the
transgene) under the severe drought stress conditions, whereas the potential
yield of transgenic plants under normal growth conditions was not affected.
Chymotrypsin-inhibitor activity assay showed that the crude protein of the
positive transgenic plants had stronger inhibitory activity than the negative
control. Transgenic plants had less decrease of total proteins than the wild type
under drought stress. Taken together, these data indicate that OCPI1 might
potentially be useful in the genetic improvement of drought resistance in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17221232 [PubMed - indexed for MEDLINE]

845: Plant Cell Rep. 2007 Jun;26(6):783-9. Epub 2007 Jan 13.

Generation of transgenic Lolium temulentum plants by Agrobacterium
tumefaciens-mediated transformation.

Ge Y, Cheng X, Hopkins A, Wang ZY.

Forage Improvement Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble
Parkway, Ardmore, OK 73401, USA.

Lolium temulentum L. (Darnel ryegrass) has been proposed to be used as a model
species for functional genomics studies in forage and turf grasses, because it is
a self-fertile, diploid species with a short life cycle and is closely related to
other grasses. Embryogenic calluses were induced from mature embryos of a double
haploid line developed through anther culture. The calluses were broken up into
small pieces and used for Agrobacterium tumefaciens-mediated transformation. A.
tumefaciens strain EHA105 harboring pCAMBIA1301 and pCAMBIA1305.2 vectors were
used to infect embryogenic callus pieces. Hygromycin was used as a selection
agent in stable transformation experiments. Hygromycin resistant calluses were
obtained after 4-6 weeks of selection and transgenic plants were produced in
10-13 weeks after Agrobacterium-mediated transformation. Fertile plants were
readily obtained after transferring the transgenics to the greenhouse. Transgenic
nature of the regenerated plants was demonstrated by Polymerase chain reaction
(PCR), Southern hybridization analysis, and GUS staining. Progeny analysis showed
Mendelian inheritance of the transgenes. The transformation system provides a
valuable tool for functionality tests of candidate genes in forage and turf
grasses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17221228 [PubMed - indexed for MEDLINE]

846: Plant Cell Rep. 2007 Jun;26(6):791-804. Epub 2007 Jan 13.

Pyramiding transgenic resistance in elite indica rice cultivars against the
sheath blight and bacterial blight.

Maruthasalam S, Kalpana K, Kumar KK, Loganathan M, Poovannan K, Raja JA,
Kokiladevi E, Samiyappan R, Sudhakar D, Balasubramanian P.

Rice Transformation Laboratory, Center for Plant Molecular Biology, Department of
Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University,
Coimbatore 641003, Tamil Nadu, India.

Elite indica rice cultivars were cotransformed with genes expressing a rice
chitinase (chi11) and a thaumatin-like protein (tlp) conferring resistance to
fungal pathogens and a serine-threonine kinase (Xa21) conferring bacterial blight
resistance, through particle bombardment, with a view to pyramiding sheath blight
and bacterial blight resistance. Molecular analyses of putative transgenic lines
by polymerase chain reaction, Southern Blot hybridization, and Western Blotting
revealed stable integration and expression of the transgenes in a few independent
transgenic lines. Progeny analyses showed the stable inheritance of transgenes to
their progeny. Coexpression of chitinase and thaumatin-like protein in the
progenies of a transgenic Pusa Basmati1 line revealed an enhanced resistance to
the sheath blight pathogen, Rhizoctonia solani, as compared to that in the lines
expressing the individual genes. A transgenic Pusa Basmati1 line pyramided with
chi11, tlp, and Xa21 showed an enhanced resistance to both sheath blight and
bacterial blight.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17221225 [PubMed - indexed for MEDLINE]

847: ScientificWorldJournal. 2007 Jan 10;7:31-46.

Mechanisms of action of probiotics in intestinal diseases.

O'Hara AM, Shanahan F.

Alimentary Pharmabiotic Centre, University College Cork, National University of
Ireland, Ireland. a.ohara@ucc.ie

Intestinal microbiota is a positive health asset that exerts a conditioning
effect on intestinal homeostasis. Resident bacteria deliver regulatory signals to
the epithelium and instruct mucosal immune responses. Recent research has
revealed a potential therapeutic role for the manipulation of the microbiota and
exploitation of host-microbial signalling pathways in the maintenance of human
health and treatment of various mucosal disorders. A variety of pharmabiotic
strategies, such as the use of specific members of the microbiota, their surface
components, or metabolites, as well as genetically modified commensal bacteria,
are being investigated for their ability to enhance the beneficial components of
the microbiota. It is clear that engagement with host cells is central to
pharmabiotic action, and several strain-specific mechanisms of action have been
elucidated. However, the molecular details underpinning these mechanisms remain
almost entirely unknown. Understanding how pharmabiotics exert their beneficial
effects is critical for the establishment of definitive selection criteria for
certain pharmabiotic strategies for specific clinical conditions. Scientifically
accredited evidence of efficacy and studies to elucidate the molecular mechanisms
of host-microbiota interactions are needed to lend credence to the use of
pharmabiotic strategies in clinical medicine.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17221140 [PubMed - indexed for MEDLINE]

848: Plant Physiol. 2007 Mar;143(3):1362-71. Epub 2007 Jan 12.

Auxin biosynthesis by the YUCCA genes in rice.

Yamamoto Y, Kamiya N, Morinaka Y, Matsuoka M, Sazuka T.

Bioscience and Biotechnology Center, Nagoya University Chikusa, Nagoya Aichi,
464-8601, Japan.

Although indole-3-acetic acid (IAA), the predominant auxin in plants, plays a
critical role in various plant growth and developmental processes, its
biosynthesis and regulation have not been clearly elucidated. To investigate the
molecular mechanisms of IAA synthesis in rice (Oryza sativa), we identified seven
YUCCA-like genes (named OsYUCCA1-7) in the rice genome. Plants overexpressing
OsYUCCA1 exhibited increased IAA levels and characteristic auxin overproduction
phenotypes, whereas plants expressing antisense OsYUCCA1 cDNA displayed defects
that are similar to those of rice auxin-insensitive mutants. OsYUCCA1 was
expressed in almost all of the organs tested, but its expression was restricted
to discrete areas, including the tips of leaves, roots, and vascular tissues,
where it overlapped with expression of a beta-glucuronidase reporter gene
controlled by the auxin-responsive DR5 promoter. These observations are
consistent with an important role for the rice enzyme OsYUCCA1 in IAA
biosynthesis via the tryptophan-dependent pathway.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17220367 [PubMed - indexed for MEDLINE]

849: Transgenic Res. 2007 Dec;16(6):703-12. Epub 2007 Jan 12.

A passage through in vitro culture leads to efficient production of marker-free
transgenic plants in Brassica juncea using the Cre-loxP system.

Arumugam N, Gupta V, Jagannath A, Mukhopadhyay A, Pradhan AK, Burma PK, Pental D.

Centre for Genetic Manipulation of Crop Plants, University of Delhi South Campus,
Benito Juarez Road, New Delhi 110 021, India.

The Cre-loxP site-specific recombination system was deployed for removal of
marker genes from Brassica juncea (Indian mustard). Excision frequencies,
monitored by removal of nptII or gfp genes in F(1) plants of crosses between LOX
and CRE lines, were high in quiescent, differentiated somatic tissues but
extremely poor in the meristematic regions (and consequently the germinal cells)
thus preventing identification and selection of marker-free transgenic events
which are devoid of both the marker gene as well as the cre gene, in F(2)
progeny. We show that a passage through in vitro culture of F(1 )leaf explants
allows efficient development of marker-free transgenics in the F(2) generation
addressing current limitations associated with efficient use of the Cre/loxP
technology for marker gene removal.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17219247 [PubMed - indexed for MEDLINE]

850: Plant Cell Rep. 2007 Jul;26(7):1097-110. Epub 2007 Jan 12.

Isolation of cold stress-responsive genes in the reproductive organs, and
characterization of the OsLti6b gene from rice (Oryza sativa L.).

Kim SH, Kim JY, Kim SJ, An KS, An G, Kim SR.

Department of Life Science, Sogang University, Seoul 121-742, Korea.

During their reproductive stage, rice crops often are exposed to cold stress,
which leads to sterility and reduced yields. To understand the cold response
mechanism at that stage, we used an mRNA differential display method to isolate
cold-responsive genes from pre-anthesis flowers. Approximately 5,000 transcripts
were identified here, of which 123 were found to be displayed differentially
between the control (30 degrees C) and cold-treated (12 degrees C) flowers. Among
them, 26 were analyzed by northern analysis; 8 of those clones were confirmed as
cold-responsive. OsLti6b, encoding a hydrophobic protein homologous to
Arabidopsis RCI2, was analyzed in detail. RNA blot analysis revealed that its
transcript is increased by cold, salt, drought, or ABA treatments. In situ
hybridization indicated that this transcript is highly accumulated in the ovaries
and stamens of cold-treated flowers, particularly in the anther walls and
vascular tissues of the filaments. Over-expression of OsLti6b increased cold
tolerance as revealed by seedling wilting rates and ion leakages of mature
leaves, demonstrating that the extent of the tolerance correlates well with its
expression level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17219102 [PubMed - indexed for MEDLINE]

851: Science. 2007 Jan 12;315(5809):182-3.

AGBIOTECH. GM technology develops in the developing world.

Sinha G.

Publication Types:
News

PMID: 17218505 [PubMed - indexed for MEDLINE]

852: Plant J. 2007 Feb;49(3):442-51. Epub 2007 Jan 1.

Single-stranded DNA binding factor AtWHY1 modulates telomere length homeostasis
in Arabidopsis.

Yoo HH, Kwon C, Lee MM, Chung IK.

Department of Biology and Molecular Aging Research Center, Yonsei University,
Seoul 120-749, Korea.

Telomere homeostasis, a process that is essential for the maintenance of
chromosome integrity, is regulated by telomerase and a collection of associated
proteins. By mass spectrometry we have identified a new telomeric protein encoded
by the AtWHY1 (Arabidopsis thaliana Whirly 1) gene in Arabidopsis. AtWHY1
specifically binds the single-stranded plant telomeric DNA sequences, but not
double-stranded telomeric DNA. To gain insights into the function of AtWHY1 in
telomere biogenesis, we have identified two Arabidopsis lines harboring T-DNA
insertions in AtWHY1. These lines exhibit neither growth nor developmental
defects. However, AtWHY1-deficient plants show a steady increase in the length of
telomere tracts over generations. This telomere elongation is correlated with a
significant increase in telomerase activity. On the contrary, transgenic plants
expressing AtWHY1 show a decreased telomerase activity and shortened telomeres.
The evidence presented here indicates that AtWHY1 is a new family of telomere
end-binding proteins that plays a role in regulating telomere-length homeostasis
in Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17217467 [PubMed - indexed for MEDLINE]

853: Plant J. 2007 Feb;49(4):592-606. Epub 2007 Jan 8.

ABA induction of miR159 controls transcript levels of two MYB factors during
Arabidopsis seed germination.

Reyes JL, Chua NH.

Laboratory of Plant Molecular Biology, The Rockefeller University, 1230 York
Avenue, New York, NY 10021, USA.

Upon seed imbibition, abscisic acid (ABA) levels decrease to allow embryos to
germinate and develop into seedlings. However, under abiotic stress conditions,
ABA levels remain high, and growth and development are arrested. Several
transcription factors, including abscisic acid-insensitive (ABI)3 and ABI5, are
known to control this developmental checkpoint. Here, we show that, in
germinating Arabidopsis thaliana seeds, ABA induces the accumulation of microRNA
159 (miR159) in an ABI3-dependent fashion, and miRNA159 mediates cleavage of
MYB101 and MYB33 transcripts in vitro and in vivo. The two MYB transcription
factors function as positive regulators of ABA responses, as null mutants of
myb33 and myb101 show hyposensitivity to the hormone. Consistent with this,
miR159 over-expression suppresses MYB33 and MYB101 transcript levels and renders
plants hyposensitive to ABA, whereas transgenic plants over-expressing
cleavage-resistant forms of MYB33 and MYB101 are hypersensitive, as are plants
over-expressing the Turnip mosaic virus (TuMV) P1/HC-Pro viral protein that is
known to inhibit miRNA function. Our results suggest that ABA-induced
accumulation of miR159 is a homeostatic mechanism to direct MYB33 and MYB101
transcript degradation to desensitize hormone signaling during seedling stress
responses.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 17217461 [PubMed - indexed for MEDLINE]

854: Plant J. 2007 Feb;49(4):607-18. Epub 2007 Jan 8.

Early events in the pathogenicity of Pseudomonas syringae on Nicotiana
benthamiana.

Hann DR, Rathjen JP.

The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.

Conserved microbial molecules known as PAMPs (pathogen-associated molecular
patterns) elicit defence responses in plants through extracellular receptor
proteins. One important PAMP is the flagellin protein derived from motile
bacteria. We show here that the solanaceous species Nicotiana benthamiana
perceives the flagellin proteins of both pathogenic and non-host species of
Pseudomonas syringae. The response to flagellin required a gene closely related
to that encoding the Arabidopsis thaliana flagellin receptor that we designated
NbFls2. In addition, silencing of NbFls2 led to increased growth of compatible,
non-host and non-pathogenic strains of P. syringae. Thus, flagellin perception
restricts growth of P. syringae strains on N. benthamiana. Pathogenic bacteria
secrete effector proteins into the plant cell to enhance virulence. We tested the
ability of several unrelated effectors to suppress PAMP-mediated defences. The
effector proteins AvrPto and AvrPtoB, but not AvrRps4, suppressed all responses
tested including the hypersensitive response induced by non-host flagellins and
the oomycete elicitor INF1. Strikingly, transient expression of avrPto or avrPtoB
stimulated the growth of non-pathogenic Agrobacterium tumefaciensin planta,
suggesting that multiplication of this species is also restricted by PAMP
perception. Unexpectedly, AvrPtoB but not AvrPto required the defence-associated
genes Rar1, Sgt1 and Eds1 for suppression. This observation separates the
respective mechanisms of the two effectors, and suggests that AvrPtoB may target
the defence machinery directly for its suppressive effect.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17217460 [PubMed - indexed for MEDLINE]

855: Transgenic Res. 2007 Oct;16(5):629-43. Epub 2007 Jan 11.

A wheat genomic DNA fragment reduces pollen transmission of maize transgenes by
reducing pollen viability.

Scott MP, Peterson JM, Moran DL, Sangtong V, Smith L.

USDA-ARS, Ames, IA 50011, USA. pscott@iastate.edu

A genomic DNA fragment from wheat carrying the Glu-1Dx5 gene has been shown to
exhibit reduced pollen transmission in transgenic maize. To localize the region
of the DNA fragment responsible for this reduced pollen transmission, we produced
transgenic maize plants in which the wheat genomic DNA proximal to the 1Dx5
coding sequence was replaced with the maize 27 kDa gamma-zein promoter. Like the
wheat promoter-driven Glu-1Dx5 transgene, this zein promoter-driven transgene
functioned to produce 1Dx5 in maize endosperm. However, with the zein
promoter-driven transgene, pollen transmission of the transgene loci was normal
in most self- and cross-pollinations. We concluded that the wheat genomic DNA
proximal to the wheat 1Dx5 coding sequence was required for reduced pollen
transmission of the transgene in maize. In two of four transformation events of
the wheat promoter-driven construct examined, pollen exhibited two morphological
classes. In one class, pollen was normal in morphology and displayed average
viability, and in the second, pollen was reduced in size and did not germinate on
artificial media. DNA from the transgene was detectable in mature pollen from
plants with reduced pollen transmission of transgene loci. To explain these
observations, we hypothesize that elements within the transgene construct
interfere with pollen development. We demonstrated that the wheat genomic DNA
fragment can be used to control pollen transmission of an herbicide resistance
transgene genetically linked to it. The wheat genomic DNA fragment may contain
elements that are useful for controlling pollen transmission of transgene loci in
commercial maize grain and seed production.

PMID: 17216545 [PubMed - indexed for MEDLINE]

856: Planta. 2007 Jun;226(1):99-108. Epub 2007 Jan 11.

Over-expression of rice OsAGO7 gene induces upward curling of the leaf blade that
enhanced erect-leaf habit.

Shi Z, Wang J, Wan X, Shen G, Wang X, Zhang J.

National Key Laboratory of Plant Molecular Genetics, Institute of Plant
Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese
Academy of Sciences, Shanghai , 200032, China.

High-yield cultivars are characterized by erect leaf canopies that optimize
photosynthesis and thus favor increased biomass. Upward curling of the leaf blade
(called rolled leaf) can result in enhanced erect-leaf habit, increase erect
duration and promote an overall erect leaf canopy. The rice mutant R05, induced
through transferred DNA (T-DNA) insertion, had the rolled-leaf trait. The leaves
in the wild type demonstrated natural drooping tendencies, resulting in
decreasing leaf erection indices (LEIs) during senescence at the 20th day after
flowering. Conversely, LEIs of the leaves in R05 remained high, even 20-day
post-flowering. We applied T-DNA tagging and isolated a rolled-leaf gene from
rice which, when over-expressed, could induce upward curling of the leaf blade.
This gene encodes for a protein of 1,048 amino acids including the PAZ and PIWI
conserved domains, belonging to the Argonaute (AGO) family. There are at least 18
members of the AGO family in rice. According to high-sequence conservation, the
rolled-leaf gene in rice could be orthologous to the Arabidopsis ZIP/Ago7 gene,
so we called it OsAGO7. These results provide a possible opportunity for
implementing OsAGO7 gene in crop improvement.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17216479 [PubMed - indexed for MEDLINE]

857: Nature. 2007 Jan 11;445(7124):132-3.

Out of bounds.

[No authors listed]

Publication Types:
News

PMID: 17215811 [PubMed - indexed for MEDLINE]

858: J Mol Med. 2007 Apr;85(4):405-13. Epub 2007 Jan 9.

Erratum in:
J Mol Med. 2007 Apr;85(4):421.

Copper and clioquinol treatment in young APP transgenic and wild-type mice:
effects on life expectancy, body weight, and metal-ion levels.

Schö¤fer S, Pajonk FG, Multhaup G, Bayer TA.

Department of Psychiatry, Division of Neurobiology, Saarland University, Homburg,
Germany.

There is mounting evidence that the amyloid precursor protein (APP), the key
protein in Alzheimer's disease (AD) is involved in the copper (Cu) homeostasis in
the brain. Conflicting results about the potential use of dietary Cu and
clioquinol (CQ), a known Cu chelator, have been reported using APP transgenic
mice. Previously, in vitro studies have demonstrated that CQ can act as a Cu
transporter. To analyze the potential function of CQ as a Cu transporter in vivo,
the nutritional effect of Cu and CQ was analyzed in young APP transgenic mice and
nontransgenics with food pellets containing either Cu, CQ, Cu plus CQ (Cu + CQ),
or without addition of supplements (control). The offspring were fed with
corresponding food pellets until the age of 14 weeks. We observed an increased
lethality of APP transgenics upon CQ treatment, which could be rescued by a
co-treatment with Cu. The exposure of Cu + CQ led to a modest but significant
increase in cerebral Cu levels, most likely due to an enhanced transport of CQ-Cu
complexes. In CQ or Cu + CQ treatment groups, the plasma levels of Cu, zinc, and
iron were reduced in all animals; moreover, Cu treatment alone reduced only
plasma iron levels. We conclude not only that CQ has certain toxicity but also
that the chelating effect, perhaps, plays a secondary role with respect to its
properties as an intracellular Cu transporter, thus, counteracting the supposed
therapeutic effects of CQ as an agent for chelating therapy in AD.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17211610 [PubMed - indexed for MEDLINE]

859: Plant Mol Biol. 2007 Mar;63(5):703-18. Epub 2007 Jan 9.

Interaction network of proteins associated with abiotic stress response and
development in wheat.

Tardif G, Kane NA, Adam H, Labrie L, Major G, Gulick P, Sarhan F, Lalibertö© JF.

Institut Armand-Frappier, Institut national de la recherche scientifique, 531
boulevard des Prairies, Laval, Quö©bec, Canada, H7V 1B7.

Wheat is the most widely adapted crop to abiotic stresses and considered an
excellent system to study stress tolerance in spite of its genetic complexity.
Recent studies indicated that several hundred genes are either up- or
down-regulated in response to stress treatment. To elucidate the function of some
of these genes, an interactome of proteins associated with abiotic stress
response and development in wheat was generated using the yeast two-hybrid GAL4
system and specific protein interaction assays. The interactome is comprised of
73 proteins, generating 97 interactions pairs. Twenty-one interactions were
confirmed by bimolecular fluorescent complementation in Nicotiana benthamiana. A
confidence-scoring system was elaborated to evaluate the significance of the
interactions. The main feature of this interactome is that almost all bait
proteins along with their interactors were interconnected, creating a spider
web-like structure. The interactome revealed also the presence of a "cluster of
proteins involved in flowering control" in three- and four-protein interaction
loops.This network provides a novel insight into the complex relationships among
transcription factors known to play central roles in vernalization, flower
initiation and abscisic acid signaling, as well as associations that tie abiotic
stress with other regulatory and signaling proteins. This analysis provides
useful information in elucidating the molecular mechanism associated with abiotic
stress response in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17211514 [PubMed - indexed for MEDLINE]

860: Plant Mol Biol. 2007 Mar;63(5):719-30. Epub 2007 Jan 9.

ci21A/Asr1 expression influences glucose accumulation in potato tubers.

Frankel N, Nunes-Nesi A, Balbo I, Mazuch J, Centeno D, Iusem ND, Fernie AR,
Carrari F.

Laboratorio de Fisiologö­a y Biologö­a Molecular, IFIBYNE-Conicet, Facultad de
Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires,
Argentina.

Asr genes are exclusively found in the genomes of higher plants. In many species,
this gene family is expressed under abiotic stress conditions and during fruit
ripening. The encoded proteins have nuclear localisation and consequently a
transcription factor function has been suggested. Interestingly, yeast-one-hybrid
experiments revealed that a grape ASR binds to the promoter of a hexose
transporter gene (VvHT1). However, the role of these proteins in planta is still
elusive. By using a reverse genetics approach in potato we found that
modification of Asr1 expression has no incidence on the aerial phenotype of the
plant but exerts a dramatic effect in tuber. Asr1 antisense potatoes displayed
decreased tuber fresh weight whereas Asr1 overexpressors had a diminished number
of tubers. Moreover, overexpression lines showed lower transcript levels of a
plasma membrane hexose transporter and a concomitant decrease in glucose content
in parenchyma cells of potato tubers. On the same hand glucose uptake rate was
also reduced in one of the overexpressing lines. It thus seems likely that Asr1
is involved in the control of hexose uptake in heterotrophic organs. In addition,
the transgenic plants were characterized by several other changes in steady state
metabolite levels. Results presented here support a role for ci21A/Asr1 in
glucose metabolism of potato tuber.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17211513 [PubMed - indexed for MEDLINE]

861: Nat Biotechnol. 2007 Jan;25(1):77-83.

Risk assessment of meat and milk from cloned animals.

Yang X, Tian XC, Kubota C, Page R, Xu J, Cibelli J, Seidel G Jr.

Center for Regenerative Biology and Department of Animal Science, University of
Connecticut, Storrs, Connecticut 06269-4243, USA. xiangzhong.yang@uconn.edu

Research on, and commercialization of, cloned cattle has been conducted for more
than 20 years. Early techniques relied on the physical splitting of embryos or
using embryo cells for nuclear transfer to generate cloned animals. Milk and meat
from these animals entered into the human food market with no evidence of
problems. With the advent of nuclear transfer, which enables the direct
transference and preservation of high-value meat- and milk-producing genotypes to
offspring, concerns have been raised about whether the products from somatic cell
nuclear transfer-produced animals are safe for human consumption. Studies on the
biochemical properties of food products from cloned and noncloned animals have
thus far not detected any differences. All data to date indicate no significant
differences in the measured parameters between animals created by nuclear
transfer and normally bred animals. Public acceptance of cloned animal products
depends upon forthcoming US Food and Drug Administration approval along with
convincing safety data.

Publication Types:
Review

PMID: 17211406 [PubMed - indexed for MEDLINE]

862: Nat Biotechnol. 2007 Jan;25(1):47-53.

Dolly for dinner? Assessing commercial and regulatory trends in cloned livestock.

Suk J, Bruce A, Gertz R, Warkup C, Whitelaw CB, Braun A, Oram C, Rodrö­guez-Cerezo
E, Papatryfon I.

ESRC Genomics Policy & Research Forum, University of Edinburgh, St. John's Land,
Edinburgh, Scotland.

As cloning technologies become more widely established, will products enter the
food chain sooner than regulatory agencies and the public might be prepared for?

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17211395 [PubMed - indexed for MEDLINE]

863: Nat Biotechnol. 2007 Jan;25(1):39-43.

Animal cloning and the FDA--the risk assessment paradigm under public scrutiny.

Rudenko L, Matheson JC, Sundlof SF.

Center for Veterinary Medicine, US Food and Drug Administration, Department of
Health and Human Services, 7500 Standish Place, Rockville, Maryland 20855, USA.
larisa.rudenko@fda.hhs.gov

The evidence gathered thus far--ultimately to be published in the Draft Risk
Assessment on Animal Cloning--indicates that there are no unique risks associated
with animal cloning.

Publication Types:
Review

PMID: 17211392 [PubMed - indexed for MEDLINE]

864: Nat Biotechnol. 2007 Jan;25(1):35-6; author reply 36-7.

Comment on:
Nat Biotechnol. 2005 Dec;23(12):1475-6. Nat Biotechnol. 2006 Jan;24(1):63-71.

Early-tier tests insufficient for GMO risk assessment.

Lang A, Lauber E, Darvas B.

Publication Types:
Comment
Letter

PMID: 17211390 [PubMed - indexed for MEDLINE]

865: Nat Biotechnol. 2007 Jan;25(1):33-4.

Comment on:
Nat Biotechnol. 2006 Oct;24(10):1178.

Parallel biopolitical universes.

Morris SH.

Publication Types:
Comment
Letter

PMID: 17211389 [PubMed - indexed for MEDLINE]

866: Nat Biotechnol. 2007 Jan;25(1):7-8. Epub 2007 Jan 5.

FDA's cloning report bypasses ethics, exposes European dilemma.

Vermij P.

Publication Types:
News

PMID: 17211377 [PubMed - indexed for MEDLINE]

867: Nat Biotechnol. 2007 Jan;25(1):1.

Comment in:
Nat Biotechnol. 2007 Mar;25(3):281. Nat Biotechnol. 2007 Mar;25(3):282-3.

The emperor's new clones.

[No authors listed]

If regulators conclude that food from clones poses no more risk than food from
other animals, the US and Europe could be on course for another biotech trade
war.

Publication Types:
Editorial

PMID: 17211372 [PubMed - indexed for MEDLINE]

868: Plant Cell. 2007 Jan;19(1):296-319. Epub 2007 Jan 5.

Rice SCAMP1 defines clathrin-coated, trans-golgi-located tubular-vesicular
structures as an early endosome in tobacco BY-2 cells.

Lam SK, Siu CL, Hillmer S, Jang S, An G, Robinson DG, Jiang L.

Department of Biology and Molecular Biotechnology Program, Chinese University of
Hong Kong, Shatin, New Territories, Hong Kong, China.

We recently identified multivesicular bodies (MVBs) as prevacuolar compartments
(PVCs) in the secretory and endocytic pathways to the lytic vacuole in tobacco
(Nicotiana tabacum) BY-2 cells. Secretory carrier membrane proteins (SCAMPs) are
post-Golgi, integral membrane proteins mediating endocytosis in animal cells. To
define the endocytic pathway in plants, we cloned the rice (Oryza sativa) homolog
of animal SCAMP1 and generated transgenic tobacco BY-2 cells expressing yellow
fluorescent protein (YFP)-SCAMP1 or SCAMP1-YFP fusions. Confocal
immunofluorescence and immunogold electron microscopy studies demonstrated that
YFP-SCAMP1 fusions and native SCAMP1 localize to the plasma membrane and mobile
structures in the cytoplasm of transgenic BY-2 cells. Drug treatments and
confocal immunofluorescence studies demonstrated that the punctate cytosolic
organelles labeled by YFP-SCAMP1 or SCAMP1 were distinct from the Golgi apparatus
and PVCs. SCAMP1-labeled organelles may represent an early endosome because the
internalized endocytic markers FM4-64 and AM4-64 reached these organelles before
PVCs. In addition, wortmannin caused the redistribution of SCAMP1 from the early
endosomes to PVCs, probably as a result of fusions between the two compartments.
Immunogold electron microscopy with high-pressure frozen/freeze-substituted
samples identified the SCAMP1-positive organelles as tubular-vesicular structures
at the trans-Golgi with clathrin coats. These early endosomal compartments
resemble the previously described partially coated reticulum and trans-Golgi
network in plant cells.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17209124 [PubMed - indexed for MEDLINE]

869: Appl Environ Microbiol. 2007 Mar;73(5):1622-9. Epub 2007 Jan 5.

Reduced contamination by the Fusarium mycotoxin zearalenone in maize kernels
through genetic modification with a detoxification gene.

Igawa T, Takahashi-Ando N, Ochiai N, Ohsato S, Shimizu T, Kudo T, Yamaguchi I,
Kimura M.

Plant & Microbial Metabolic Engineering Research Unit and Laboratory for
Remediation Research, Discovery Research Institute (DRI), RIKEN, 2-1 Hirosawa,
Wako, Saitama 351-0198, Japan.

Maize is subject to ear rot caused by toxigenic Aspergillus and Fusarium species,
resulting in contamination with aflatoxins, fumonisins, trichothecenes, and
zearalenone (ZEN). The trichothecene group and ZEN mycotoxins are produced by the
cereal pathogen Fusarium graminearum. A transgenic detoxification system for the
elimination of ZEN was previously developed using an egfp::zhd101 gene
(gfzhd101), encoding an enhanced green fluorescent protein fused to a
ZEN-degrading enzyme. In this study, we produced a transgenic maize line
expressing an intact copy of gfzhd101 and examined the feasibility of
transgene-mediated detoxification in the kernels. ZEN-degrading activity has been
detected in transgenic kernels during seed maturation (for a period of 6 weeks
after pollination). The level of detoxification activity was unaltered after an
additional storage period of 16 weeks at 6 degrees C. When the seeds were
artificially contaminated by immersion in a ZEN solution for 48 h at 28 degrees
C, the total amount of the mycotoxin in the transgenic seeds was uniformly
reduced to less than 1/10 of that in the wild type. The ZEN in the transgenic
maize kernels was also efficiently decontaminated under conditions of lower water
activity (aw) and temperature; e.g., 16.9 microg of ZEN was removed per gram of
seed within 48 h at an aw of 0.90 at 20 degrees C. F. graminearum infection
assays demonstrated an absence of ZEN in the transgenic maize seeds, while the
mycotoxin accumulated in wild-type kernels under the same conditions.
Transgene-mediated detoxification may offer simple solutions to the problems of
mycotoxin contamination in maize.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17209063 [PubMed - indexed for MEDLINE]

870: Plant Physiol. 2007 Mar;143(3):1347-61. Epub 2007 Jan 5.

The grapevine transcription factor VvMYBPA1 regulates proanthocyanidin synthesis
during fruit development.

Bogs J, Jaffö© FW, Takos AM, Walker AR, Robinson SP.

Commonwealth Scientific and Industrial Research Organization Plant Industry, Glen
Osmond, 5064, Australia.

Proanthocyanidins (PAs; or condensed tannins) can protect plants against
herbivores, contribute to the taste of many fruits, and act as dietary
antioxidants beneficial for human health. We have previously shown that in
grapevine (Vitis vinifera) PA synthesis involves both leucoanthocyanidin
reductase (LAR) and anthocyanidin reductase (ANR). Here we report the
characterization of a grapevine MYB transcription factor VvMYBPA1, which controls
expression of PA pathway genes including both LAR and ANR. Expression of VvMYBPA1
in grape berries correlated with PA accumulation during early berry development
and in seeds. In a transient assay, VvMYBPA1 activated the promoters of LAR and
ANR, as well as the promoters of several of the general flavonoid pathway genes.
VvMYBPA1 did not activate the promoter of VvUFGT, which encodes the
anthocyanin-specific enzyme UDP-glucose:flavonoid-3-O-glucosyltransferase,
suggesting VvMYBPA1 is specific to regulation of PA biosynthesis in grapes. The
Arabidopsis (Arabidopsis thaliana) MYB transcription factor TRANSPARENT TESTA2
(TT2) regulates PA synthesis in the seed coat of Arabidopsis. By complementing
the PA-deficient seed phenotype of the Arabidopsis tt2 mutant with VvMYBPA1, we
confirmed the function of VvMYBPA1 as a transcriptional regulator of PA
synthesis. In contrast to ectopic expression of TT2 in Arabidopsis, constitutive
expression of VvMYBPA1 resulted in accumulation of PAs in cotyledons, vegetative
meristems, leaf hairs, and roots in some of the transgenic seedlings. To our
knowledge, this is the first report of a MYB factor that controls genes of the PA
pathway in fruit, including both LAR and ANR, and this single MYB factor can
induce ectopic PA accumulation in Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17208963 [PubMed - indexed for MEDLINE]

871: Plant Physiol. 2007 Feb;143(2):720-31. Epub 2007 Jan 5.

Abscisic acid and stress signals induce Viviparous1 expression in seed and
vegetative tissues of maize.

Cao X, Costa LM, Biderre-Petit C, Kbhaya B, Dey N, Perez P, McCarty DR,
Gutierrez-Marcos JF, Becraft PW.

Department of Genetics, Development and Cell Biology, Iowa State University,
Ames, Iowa 50011, USA.

Viviparous1 (Vp1) encodes a B3 domain-containing transcription factor that is a
key regulator of seed maturation in maize (Zea mays). However, the mechanisms of
Vp1 regulation are not well understood. To examine physiological factors that may
regulate Vp1 expression, transcript levels were monitored in maturing embryos
placed in culture under different conditions. Expression of Vp1 decreased after
culture in hormone-free medium, but was induced by salinity or osmotic stress.
Application of exogenous abscisic acid (ABA) also induced transcript levels
within 1 h in a dose-dependent manner. The Vp1 promoter fused to
beta-glucuronidase or green fluorescent protein reproduced the endogenous Vp1
expression patterns in transgenic maize plants and also revealed previously
unknown expression domains of Vp1. The Vp1 promoter is active in the embryo and
aleurone cells of developing seeds and, upon drought stress, was also found in
phloem cells of vegetative tissues, including cobs, leaves, and stems. Sequence
analysis of the Vp1 promoter identified a potential ABA-responsive complex,
consisting of an ACGT-containing ABA response element (ABRE) and a coupling
element 1-like motif. Electrophoretic mobility shift assay confirmed that the
ABRE and putative coupling element 1 components specifically bound proteins in
embryo nuclear protein extracts. Treatment of embryos in hormone-free Murashige
and Skoog medium blocked the ABRE-protein interaction, whereas exogenous ABA or
mannitol treatment restored this interaction. Our data support a model for a
VP1-dependent positive feedback mechanism regulating Vp1 expression during seed
maturation.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17208960 [PubMed - indexed for MEDLINE]

872: Colloids Surf B Biointerfaces. 2007 Apr 1;55(2):159-63. Epub 2006 Dec 9.

Stable expression of foot-and-mouth disease virus protein VP1 fused with cholera
toxin B subunit in the potato (Solanum tuberosum).

He DM, Qian KX, Shen GF, Li YN, Zhang ZF, Su ZL, Shao HB.

Department of Biotechnology, College of Life Science, Zhejiang University,
Hangzhou 310027, China.

The expression vector, pBI121CTBVP1, containing the fusion of the foot and mouth
disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was
constructed by fused PCR and transferred into potato (Solanum tuberosum L.) by
Agrobacterium-mediated transformation. Transformed plants were obtained by
selecting on kanamycin-resistant medium strictly and regenerated. The transgenic
plantlets were identified by PCR, Southern-blot and the production of fused
protein was confirmed and quantified by Western-blot and ELISA assays. The
results showed that the fused genes were expressed stablely under the control of
specific-tuber patatin promoter. The expressed fused proteins have a certain
degree of immunogenicity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17208421 [PubMed - indexed for MEDLINE]

873: J Plant Physiol. 2007 Apr;164(4):514-20. Epub 2007 Jan 4.

Heterologous expression of the mutated melon ethylene receptor gene Cm-ERS1/H70A
produces stable sterility in transgenic lettuce (Lactuca sativa).

Takada K, Watanabe S, Sano T, Ma B, Kamada H, Ezura H.

Gene Research Center, Graduate School of Life and Environmental Sciences,
University of Tsukuba, Tsukuba, Ibaraki, Japan.

The mutated melon ethylene receptor gene Cm-ERS1/H70A was introduced into tobacco
and induced stable sterility in transgenic lines. This gene contains a missense
mutation that converts the His(70) residue to Ala in the melon ethylene receptor
gene Cm-ERS1. To test the applicability of this inducible sterility system to
other plants, lettuce (Lactuca sativa) was transformed with the gene using
Agrobacterium, and putative transformants containing Cm-ERS1/H70A were obtained.
Thirteen randomly selected putative transformants were grown in a growth room
under constant conditions, and seven of the lines showed sterility or
significantly reduced fertility. DNA gel blot analysis confirmed the integration
of the Cm-ERS1/H70A gene into the genomes of the putative transformants, and
RT-PCR and protein gel blot analysis confirmed the expression of Cm-ERS1/H70A
mRNA and protein in all of the transformants. Five transformants showing
sterility or reduced fertility when grown in a growth room under constant
conditions were randomly selected to be grown in an open-air greenhouse under
various environmental conditions. All five showed stable sterility under the
various conditions. These results suggest that Cm-ERS1/H70A can induce sterility
in heterologous transgenic plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207555 [PubMed - indexed for MEDLINE]

874: Biochem Biophys Res Commun. 2007 Feb 23;353(4):863-8. Epub 2006 Dec 22.

Ethylene responsive element binding protein 1 (StEREBP1) from Solanum tuberosum
increases tolerance to abiotic stress in transgenic potato plants.

Lee HE, Shin D, Park SR, Han SE, Jeong MJ, Kwon TR, Lee SK, Park SC, Yi BY, Kwon
HB, Byun MO.

Postharvest Technology Division, National Horticulture Research Institute, RDA,
Suwon 440-706, Republic of Korea.

To identify components of the plant stress signal transduction cascade and
response mechanisms, we screened plant genes using reverse Northern blot
analysis, and chose the ethylene responsive element binding protein 1 (StEREBP1)
for further characterization. To investigate its biological function in the
potato, we performed Northern blot analysis and observed enhanced levels of
transcription in response to several environmental stresses including low
temperature. In vivo targeting experiments using a green fluorescent protein
(GFP) reporter indicated that StEREBP1 localized to the nucleus of onion
epidermal cells. StEREBP1 was found to bind to GCC and DRE/CRT cis-elements and
both microarray and RT-PCR analyses indicated that overexpression of StEREBP1
induced expression of several GCC box-containing stress response genes. In
addition, overexpression of StEREBP1 enhanced tolerance to cold and salt stress
in transgenic potato plants. The results of this study suggest that StEREBP1 is a
functional transcription factor that may be involved in abiotic stress responses
in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207469 [PubMed - indexed for MEDLINE]

875: Plant Biotechnol J. 2007 Jan;5(1):146-61.

Transcriptome analysis reveals season-specific rbcS gene expression profiles in
diploid perennial ryegrass (Lolium perenne L.).

Sathish P, Withana N, Biswas M, Bryant C, Templeton K, Al-Wahb M, Smith-Espinoza
C, Roche JR, Elborough KM, Phillips JR.

Pastoral Genomics, c/o ViaLactia Biosciences (NZ) Ltd, PO Box 109185, Newmarket,
Auckland 1149, New Zealand. sathish.puthigae@vialactia.com

Perennial ryegrass (Lolium perenne L.) is a major grass species used for forage
and turf throughout the world, and gains by conventional breeding have reached a
plateau. Perennial ryegrass is an outcrossing, self-incompatible diploid (2n = 2x
= 14) with a relatively large genome (4067 Mbp/diploid genome; Evans, G.M., Rees,
H., Snell, C.L. and Sun, S. (1972) The relation between nuclear DNA amount and
the duration of the mitotic cycle. Chrom. Today, 3, 24-31). Using tissues sourced
from active pastures during the peak of the autumn, winter, spring and summer
seasons, we analysed the ryegrass transcriptome employing a Serial Analysis of
Gene Expression (SAGE) protocol, with the dual goals of understanding the
seasonal changes in perennial ryegrass gene expression and enhancing our ability
to select genes for genetic manipulation. A total of 159,002 14-mer SAGE tags was
sequenced and mapped to the perennial ryegrass DNA database, comprising
methyl-filtered (GeneThresher) and expressed sequence tag (EST) sequences. The
analysis of 14,559 unique SAGE tags, which were present more than once in our
SAGE library, revealed 964, 1331, 346 and 131 exclusive transcripts to autumn,
winter, spring and summer, respectively. Intriguingly, our analysis of the SAGE
tags revealed season-specific expression profiles for the small subunit of
ribulose-1,5-bisphosphate carboxylase (Rubisco), LprbcS. The transcript level for
LprbcS was highest in spring, and then decreased gradually between summer and
winter. Five different copies of LprbcS were revealed in ryegrass, with one
possibly producing splice variant transcripts. Two highly expressed LprbcS genes
were reported, one of which was not active in autumn. Another LprbcS gene showed
an inverse expression profile to the autumn inactive LprbcS in a manner to
compensate the expression level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207264 [PubMed - indexed for MEDLINE]

876: Plant Biotechnol J. 2007 Jan;5(1):134-45.

Expression of an engineered granule-bound Escherichia coli maltose
acetyltransferase in wild-type and amf potato plants.

Nazarian Firouzabadi F, Vincken JP, Ji Q, Suurs LC, Visser RG.

Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding,
Wageningen University, PO Box 386, 6700 AJ Wageningen, the Netherlands.

Starch is used in many industrial applications, but often requires chemical
derivatization to enhance its properties before use. In particular, the stability
of starch polymers in solution is improved by acetylation. A drawback of this
treatment is the use of pollutant chemicals. A biological alternative to chemical
derivatization was investigated by the expression of an amyloplast-targeted
Escherichia coli maltose acetyltransferase (MAT) gene in tubers of wild-type
(Kardal) and mutant amylose-free (amf) potato plants. MAT was expressed as such,
or fused to the N- or C-terminus of a non-catalytic starch-binding domain (SBD)
to target the starch granule. Starch granules derived from transgenic plants were
found to contain acetyl groups, although their content was low, opening up an
avenue to move away from the post-harvest chemical derivatization of starch. MAT
inside starch granules was found to be active post-harvest when supplied with
acetyl-coenzyme A and glucose or maltose, but it did not acetylate starch
polymers in vitro. Starch granules from transformants in which MAT alone was
expressed also showed MAT activity, indicating that MAT is accumulated in starch
granules, and has affinity for starch by itself. Furthermore, starch granule
morphology was altered, and fusion proteins containing MAT and SBD seemed to have
a higher affinity for starch granules than two appended SBDs. These results are
discussed against the background of the quaternary structure of MAT.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207263 [PubMed - indexed for MEDLINE]

877: Plant Biotechnol J. 2007 Jan;5(1):109-17.

Doubled sugar content in sugarcane plants modified to produce a sucrose isomer.

Wu L, Birch RG.

Botany Department - SIB, The University of Queensland, Brisbane, Qld 4072,
Australia.

Sucrose is the feedstock for more than half of the world's fuel ethanol
production and a major human food. It is harvested primarily from sugarcane and
beet. Despite attempts through conventional and molecular breeding, the stored
sugar concentration in elite sugarcane cultivars has not been increased for
several decades. Recently, genes have been cloned for bacterial isomerase enzymes
that convert sucrose into sugars which are not metabolized by plants, but which
are digested by humans, with health benefits over sucrose. We hypothesized that
an appropriate sucrose isomerase (SI) expression pattern might simultaneously
provide a valuable source of beneficial sugars and overcome the sugar yield
ceiling in plants. The introduction of an SI gene tailored for vacuolar
compartmentation resulted in sugarcane lines with remarkable increases in total
stored sugar levels. The high-value sugar isomaltulose was accumulated in storage
tissues without any decrease in stored sucrose concentration, resulting in up to
doubled total sugar concentrations in harvested juice. The lines with enhanced
sugar accumulation also showed increased photosynthesis, sucrose transport and
sink strength. This remarkable step above the former ceiling in stored sugar
concentration provides a new perspective into plant source-sink relationships,
and has substantial potential for enhanced food and biofuel production.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207261 [PubMed - indexed for MEDLINE]

878: Plant Biotechnol J. 2007 Jan;5(1):84-92.

Endosperm tissue is good production platform for artificial recombinant proteins
in transgenic rice.

Takaiwa F, Takagi H, Hirose S, Wakasa Y.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, Kannondai 2-1-2, Tsukuba Ibaraki 305-8602, Japan.
takaiwa@nias.affrc.go.jp

Transgenic rice plants expressing 7Crp peptide were generated by
Agrobacterium-mediated transformation. The 7Crp peptide is the hybrid peptide of
seven major human T-cell epitopes derived from Japanese cedar pollen allergens
Cry j 1 and Cry j 2. When the 7Crp gene was expressed under the control of the
rice AGPase large subunit or maize ubiquitin-1 promoters, it could only be
detected in the endosperm of rice seed, although high levels of RNA transcript
were observed in the leaf, stem, and seed embryo. It was demonstrated by confocal
and electron microscopy analysis that the 7Crp peptide was mainly localized in
the endoplasmic reticulum-derived protein bodies, designated protein body I
(PB-I). Our results indicate that rice endosperm tissue has advantage over other
tissues as a production platform for foreign recombinant proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207259 [PubMed - indexed for MEDLINE]

879: Plant Biotechnol J. 2007 Jan;5(1):60-8.

Varietal effects of eight paired lines of transgenic Bt maize and near-isogenic
non-Bt maize on soil microbial and nematode community structure.

Griffiths BS, Heckmann LH, Caul S, Thompson J, Scrimgeour C, Krogh PH.

Environment Plant Interactions Programme, Scottish Crop Research Institute,
Invergowrie, Dundee DD2 5DA, UK. bryan.griffiths@scri.ac.uk

A glasshouse experiment was undertaken to provide baseline data on the variation
between conventional maize (Zea mays L.) varieties and genetically modified maize
plants expressing the insecticidal Bacillus thuringiensis protein (Bt, Cry1Ab).
The objective was to determine whether the variation in soil parameters under a
range of conventional maize cultivars exceeded the differences between Bt and
non-Bt maize cultivars. Variations in plant growth parameters (shoot and root
biomass, percentage carbon, percentage nitrogen), Bt protein concentration in
shoots, roots and soil, soil nematode abundance and soil microbial community
structure were determined. Eight paired varieties (i.e. varieties genetically
modified to express Bt protein and their near-isogenic control varieties) were
investigated, together with a Bt variety for which no near-isogenic control was
available (NX3622, a combined transformant expressing both Bt and herbicide
tolerance) and a conventional barley (Hordeum vulgare L.) variety which was
included as a positive control. The only plant parameter which showed a
difference between Bt varieties and near-isogenic counterparts was the shoot
carbon to nitrogen ratio; this was observed for only two of the eight varieties,
and so was not attributable to the Bt trait. There were no detectable differences
in the concentration of Bt protein in plant or soil with any of the Bt-expressing
varieties. There were significant differences in the abundance of soil nematodes,
but this was not related to the Bt trait. Differences in previously published
soil nematode studies under Bt maize were smaller than these varietal effects.
Soil microbial community structure, as determined by phospholipid fatty acid
(PLFA) analysis, was strongly affected by plant growth stage but not by the Bt
trait. The experimental addition of purified Cry1Ab protein to soil confirmed
that, at ecologically relevant concentrations, there were no measurable effects
on microbial community structure.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207257 [PubMed - indexed for MEDLINE]

880: Plant Biotechnol J. 2007 Jan;5(1):50-9.

Therapeutic effectiveness of orally administered transgenic low-alkaloid tobacco
expressing human interleukin-10 in a mouse model of colitis.

Menassa R, Du C, Yin ZQ, Ma S, Poussier P, Brandle J, Jevnikar AM.

Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food
Canada, 1391 Sandford St., London, ON, Canada, N5V 4T3.

Inflammatory bowel disease (IBD) represents a spectrum of diseases in which
inflammation leads to acute and chronic gut injury. It is a growing health issue
for which no cure exists. The pathogenesis is multifactorial with links to
infectious and environmental events that trigger disease in genetically
predisposed individuals. Treatment of the two major forms of IBD, Crohn's disease
and ulcerative colitis, involves the reduction of inflammation with toxic
immunosuppressive drugs or blocking of the pro-inflammatory effects of tumour
necrosis factor-alpha (TNF-alpha) with antibodies. Here, we show that the oral
administration of transgenic low-alkaloid tobacco expressing the
contra-inflammatory cytokine human interleukin-10 (hIL-10) reduces the severity
of colitis by down-regulating TNF-alpha expression directly at the sites of
inflammation in IBD-susceptible IL-10(-/-) mice. hIL-10 expressed in plants is
biologically active and displays resistance to gastrointestinal degradation.
Dietary supplementation with plant tissue delivering up to 9 microg of hIL-10
daily for 4 weeks was well tolerated by treated mice. Gut histology was
significantly improved relative to controls (P = 0.002), and was correlated with
a decrease in small bowel TNF-alpha mRNA levels and an increase in IL-2 and
IL-1beta mRNA levels. Transgenic plants expressing IL-10 to directly attenuate
TNF-alpha expression at sites of inflammation in the gut may become a useful new
approach in the luminal therapy of IBD.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17207256 [PubMed - indexed for MEDLINE]

881: Biotechnol Lett. 2007 Apr;29(4):641-5. Epub 2007 Jan 6.

Transient co-expression of post-transcriptional gene silencing suppressors and
beta-glucuronidase in harvested lettuce leaf tissue does not improve recombinant
protein accumulation in planta.

Simmons CW, VanderGheynst JS.

Department of Biological and Agricultural Engineering, University of California,
Davis, One Shields Avenue, Davis, CA 95616, USA.

Agrobacterium-mediated gene transfer was used to co-express three virus-derived
post-transcriptional gene silencing (PTGS) suppressors, P19 from tomato bushy
stunt virus and two species of helper component proteinase (HcPro) from tobacco
etch virus (TEV) and turnip mosaic virus, with beta-glucuronidase (GUS) in
harvested lettuce leaf tissue to investigate whether GUS accumulation increases
in the presence of PTGS suppressors. Co-expression incubations were 3-5 days at 4
and 22 degrees C. GUS activity and leaf viability were measured after incubation.
Co-expression of PTGS suppressors did not elevate GUS expression levels. Under
certain incubation conditions, co-expression of TEV HcPro significantly lowered
transient GUS expression and was detrimental to leaf viability, suggesting that
expression of PTGS silencers may have a negative effect on transient expression
levels that outweighs any effects of PTGS suppression in harvested leaf tissues.

PMID: 17206371 [PubMed - indexed for MEDLINE]

882: Biotechnol Lett. 2007 Apr;29(4):659-67. Epub 2007 Jan 6.

Expression of xylanase with high specific activity from Streptomyces
olivaceoviridis A1 in transgenic potato plants (Solanum tuberosum L.).

Yang P, Wang Y, Bai Y, Meng K, Luo H, Yuan T, Fan Y, Yao B.

Department of Microbial Engineering, Feed Research Institute, Chinese Academy of
Agricultural Sciences, Beijing, China.

The gene, xynB, from Streptomyces olivaceoviridis A1 encoding xylanase, XYNB,
with a high specific activity for xylan, was transformed into potato (Solanum
tuberosum L.) by Agrobacterium tumefaciens. The integration of xynB into genomic
DNA was confirmed by PCR and reverse transcriptase-PCR. The gene was expressed
under the control of a constitutive double cauliflower mosaic virus (CaMV) 35S
promoter. Both SDS-PAGE and western blot analysis showed high levels of
expression of the 21 kDa and 31 kDa XYNB proteins in transgenic potato plants
transformed by the binary vectors pBinXy and signal peptide contained pBinSPXy,
respectively. The recombinant XYNB protein was present at up to 5% of total
soluble leaf protein in the cytoplasm. In transgenic leaf and tuber extracts,
xylanase activity was up to 87 micromol min(-1) g(-1) fresh leaf (9.7 micromol
min(-1) mg(-1) total soluble protein). The xylanase was stable at 60 degrees C
and 70 degrees C in buffers (pH 5.2) for 5 min. Furthermore, the xylanase
enzymatic activity remained virtually unchanged over several generations of
potato. These results demonstrate that the transgenic potato can be used to
produce recombinant xylanase with high specific enzyme activity and can
potentially be an alternative to present-day xylanase additives to animal feed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17206370 [PubMed - indexed for MEDLINE]

883: Plant Cell Rep. 2007 Jun;26(6):765-72. Epub 2007 Jan 5.

The use of the phosphomannose isomerase gene as alternative selectable marker for
Agrobacterium-mediated transformation of flax (Linum usitatissimum).

Lamblin F, Aimö© A, Hano C, Roussy I, Domon JM, Van Droogenbroeck B, Lainö© E.

Laboratoire de Biologie des Ligneux et des Grandes Cultures, UPRES EA 1207,
Antenne Scientifique Universitaire de Chartres, Chartres, France.
frederic.lamblin@univ-orleans.fr

In order to meet the future requirement of using non-antibiotic resistance genes
for the production of transgenic plants, we have adapted the selectable marker
system PMI/mannose to be used in Agrobacterium-mediated transformation of flax
(Linum usitatissimum L.) cv. Barbara. The Escherichia coli pmi gene encodes a
phosphomannose isomerase (E.C. 5.1.3.8) that converts mannose-6-phosphate, an
inhibitor of glycolysis, into fructose-6-phosphate (glycolysis intermediate). Its
expression in transformed cells allows them to grow on mannose-selective medium.
The Agrobacterium tumefaciens strain GV3101 (pGV2260) harbouring the binary
vector pNOV2819 that carries the pmi gene under the control of the Cestrum yellow
leaf curling virus constitutive promoter was used for transformation experiments.
Transgenic flax plants able to root on mannose-containing medium were obtained
from hypocotyl-derived calli that had been selected on a combination of 20 g
L(-1) sucrose and 10 g L(-1) mannose. Their transgenic state was confirmed by PCR
and Southern blotting. Transgene expression was detected by RT-PCR in leaves,
stems and roots of in vitro grown primary transformants. The mean transformation
efficiency of 3.6%, that reached 6.4% in one experiment was comparable to that
obtained when using the nptII selectable marker on the same cultivar. The ability
of T1 seeds to germinate on mannose-containing medium confirmed the Mendelian
inheritance of the pmi gene in the progeny of primary transformants. These
results indicate that the PMI/mannose selection system can be successfully used
for the recovery of flax transgenic plants under safe conditions for human health
and the environment.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17205337 [PubMed - indexed for MEDLINE]

884: New Phytol. 2007;173(2):346-53.

Dramatic reduction of crop-to-crop gene flow within a short distance from
transgenic rice fields.

Rong J, Lu BR, Song Z, Su J, Snow AA, Zhang X, Sun S, Chen R, Wang F.

Ministry of Education Key Laboratory for Biodiversity and Ecological Engineering,
Institute of Biodiversity Science, Fudan University, Handan Road 220, Shanghai
200433, China.

Genetically modified (GM) rice with enhanced agronomic traits and pharmaceutical
uses are ready for widespread adoption. Little is known about isolation
requirements for achieving stringent transgene confinement in rice. To
investigate the extent of pollen-mediated crop-to-crop transgene flow, we
conducted a field experiment with four plot-size treatments of adjacent GM and
nonGM rice (Oryza sativa) in China. Three insect-resistant GM rice (Bt/CpTI) and
nonGM isogenic lines were used in the study. The hygromycin-resistance transgene
(hpt) marker was used to screen seeds from the nonGM rice rows at different
distance intervals from GM rice plots. Based on the examination of > 2.1 million
germinated seeds, we found a dramatic reduction in transgene frequencies with
increasing distance from the GM crop, ranging from c. 0.28% at 0.2 m to < 0.01%
at 6.2 m. In addition, different plot size did not significantly affect the
frequencies of gene flow. In conclusion, pollen-mediated crop-to-crop transgene
flow in rice can be maintained at negligible levels with short spatial isolation.
The model can also be applied to other crops with self- and wind-pollination.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17204081 [PubMed - indexed for MEDLINE]

885: EMBO J. 2007 Jan 24;26(2):448-58. Epub 2007 Jan 4.

Nutrient starvation promotes condensin loading to maintain rDNA stability.

Tsang CK, Li H, Zheng XS.

Department of Pharmacology, Robert Wood Johnson Medical School, Piscataway, NJ
08854, USA.

Nutrient starvation or rapamycin treatment, through inhibition of target of
rapamycin, causes condensation of ribosomal DNA (rDNA) array and nucleolar
contraction in budding yeast. Here we report that under such conditions,
condensin is rapidly relocated into the nucleolus and loaded to rDNA tandem
repeats, which is required for rDNA condensation. Rpd3-dependent histone
deacetylation is necessary and sufficient for condensin's relocalization and
loading to rDNA array, suggesting that histone modification plays a regulatory
role for condensin targeting. Rapamycin independently, yet coordinately, inhibits
rDNA transcription and promotes condensin loading to rDNA array. Unexpectedly, we
found that inhibition of rDNA transcription in the absence of condensin loading
leads to rDNA instability. Our data suggest that enrichment of condensin prevents
rDNA instability during nutrient starvation. Together, these observations unravel
a novel role for condensin in the maintenance of regional genomic stability.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 17203076 [PubMed - indexed for MEDLINE]

886: J Virol. 2007 Mar;81(6):2980-94. Epub 2007 Jan 3.

A structured viroid RNA serves as a substrate for dicer-like cleavage to produce
biologically active small RNAs but is resistant to RNA-induced silencing
complex-mediated degradation.

Itaya A, Zhong X, Bundschuh R, Qi Y, Wang Y, Takeda R, Harris AR, Molina C,
Nelson RS, Ding B.

Department of Plant Cellular and Molecular Biology and Plant Biotechnology
Center, Ohio State University, 207 Rightmire Hall, 1060 Carmack Road, Columbus,
OH 43210, USA.

RNA silencing is a potent means of antiviral defense in plants and animals. A
hallmark of this defense response is the production of 21- to 24-nucleotide viral
small RNAs via mechanisms that remain to be fully understood. Many viruses encode
suppressors of RNA silencing, and some viral RNAs function directly as silencing
suppressors as counterdefense. The occurrence of viroid-specific small RNAs in
infected plants suggests that viroids can trigger RNA silencing in a host,
raising the question of how these noncoding and unencapsidated RNAs survive
cellular RNA-silencing systems. We address this question by characterizing the
production of small RNAs of Potato spindle tuber viroid (srPSTVds) and
investigating how PSTVd responds to RNA silencing. Our molecular and biochemical
studies provide evidence that srPSTVds were derived mostly from the secondary
structure of viroid RNAs. Replication of PSTVd was resistant to RNA silencing,
although the srPSTVds were biologically active in guiding RNA-induced silencing
complex (RISC)-mediated cleavage, as shown with a sensor system. Further analyses
showed that without possessing or triggering silencing suppressor activities, the
PSTVd secondary structure played a critical role in resistance to RISC-mediated
cleavage. These findings support the hypothesis that some infectious RNAs may
have evolved specific secondary structures as an effective means to evade RNA
silencing in addition to encoding silencing suppressor activities. Our results
should have important implications in further studies on RNA-based mechanisms of
host-pathogen interactions and the biological constraints that shape the
evolution of infectious RNA structures.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17202210 [PubMed - indexed for MEDLINE]

887: Rapid Commun Mass Spectrom. 2007;21(3):319-28.

Mass spectrometric detection of CP4 EPSPS in genetically modified soya and maize.

Ocaö±a MF, Fraser PD, Patel RK, Halket JM, Bramley PM.

Centre for Chemical and Bioanalytical Sciences, Royal Holloway, University of
London, Egham TW20 0EX, UK.

The potential of protein fractionation hyphenated to mass spectrometry (MS) to
detect and characterize the transgenic protein present in Roundup Ready soya and
maize has been investigated. Genetically modified (GM) soya and maize contain the
5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene from Agrobacterium
tumefaciens CP4, which confers resistance to the herbicide glyphosate. The GM
soya and maize proteomes were fractionated by gel filtration, anion-exchange
chromatography and sodium dodecyl sulfate/polyacrylamide gel electrophoresis
(SDS-PAGE) prior to MS. This facilitated detection of a tryptic peptide map of
CP4 EPSPS by matrix-assisted laser desorption/ionization time-of-flight
(MALDI-TOF) MS and nanoelectrospray ionization quadrupole time-of-flight
(nanoESI-QTOF) MS. Subsequently, sequence information from the CP4 EPSPS tryptic
peptides was obtained by nanoESI-QTOF MS/MS. The identification was accomplished
in 0.9% GM soya seeds, which is the current EU threshold for food-labeling
requirements. Copyright 2007 John Wiley & Sons, Ltd.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17200978 [PubMed - indexed for MEDLINE]

888: J Agric Food Chem. 2007 Jan 10;55(1):15-24.

Event-specific quantitative detection of nine genetically modified maizes using
one novel standard reference molecule.

Yang L, Guo J, Pan A, Zhang H, Zhang K, Wang Z, Zhang D.

SJTU-SIBS-PSU Joint Center for Life Sciences, School of Life Science and
Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai
200240, People's Republic of China.

With the development of genetically modified organism (GMO) detection techniques,
the Polymerase Chain Reaction (PCR) technique has been the mainstay for GMO
detection, and real-time PCR is the most effective and important method for GMO
quantification. An event-specific detection strategy based on the unique and
specific integration junction sequences between the host plant genome DNA and the
integrated gene is being developed for its high specificity. This study
establishes the event-specific detection methods for TC1507 and CBH351 maizes. In
addition, the event-specific TaqMan real-time PCR detection methods for another
seven GM maize events (Bt11, Bt176, GA21, MON810, MON863, NK603, and T25) were
systematically optimized and developed. In these PCR assays, the fluorescent
quencher, TAMRA, was dyed on the T-base of the probe at the internal position to
improve the intensity of the fluorescent signal. To overcome the difficulties in
obtaining the certified reference materials of these GM maizes, one novel
standard reference molecule containing all nine specific integration junction
sequences of these GM maizes and the maize endogenous reference gene, zSSIIb, was
constructed and used for quantitative analysis. The limits of detection of these
methods were 20 copies for these different GM maizes, the limits of quantitation
were about 20 copies, and the dynamic ranges for quantification were from 0.05 to
100% in 100 ng of DNA template. Furthermore, nine groups of the mixed maize
samples of these nine GM maize events were quantitatively analyzed to evaluate
the accuracy and precision. The accuracy expressed as bias varied from 0.67 to
28.00% for the nine tested groups of GM maize samples, and the precision
expressed as relative standard deviations was from 0.83 to 26.20%. All of these
indicated that the established event-specific real-time PCR detection systems and
the reference molecule in this study are suitable for the identification and
quantification of these GM maizes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17199308 [PubMed - indexed for MEDLINE]

889: Mol Biol Rep. 2007 Mar;34(1):61-7. Epub 2006 Dec 30.

Optimization of wheat co-transformation procedure with gene cassettes resulted in
an improvement in transformation frequency.

Yao Q, Cong L, He G, Chang J, Li K, Yang G.

China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory, College of
Life Science and Technology, Huazhong University of Science and Technology,
430074, Luoyu Road 1037, Wuhan, Hubei, People's Republic of China.

Genetic manipulation using gene cassettes was applied to the elite wheat variety
EM12 via particle bombardment, which allows an improvement in transformation
frequency. We simultaneously transferred to wheat immature embryos with two
non-linked genes, gus and bar, on either separate gene cassettes or one plasmid.
The linear gene cassettes were excised and purified by restriction digestion of
the plasmid, and consisted of promoters, open reading frames and terminators. No
difference was observed in GUS transient expression of between gene cassettes and
single whole plasmid. However, the stable transformation frequency was
significantly increased to 1.1% using gene cassettes, compared with 0.4% when
using single plasmid. Procedures of the efficient co-transformation with gene
cassettes were developed. Factors influencing on the transformation frequency
were also studied in order to optimize the procedure. These were acceleration
pressure, target distance, gold particle size, the quantity ratio of gene
cassettes and the age of target explants. Based on the transient and stable
expression of the gus gene cassettes, optimization of transformation parameters
improved the reproducibility of transformation in the elite wheat variety.

Publication Types:
Evaluation Studies

PMID: 17195929 [PubMed - indexed for MEDLINE]

890: J Econ Entomol. 2006 Dec;99(6):2164-70.

Decreased whorl and ear damage in nine mon810 Bacillus thuringiensis
(Bt)-transgenic corn hybrids compared with their non-Bt counterparts.

Chilcutt CF, Odvody GN, Correa JC, Remmers J, Parker RD.

Department of Entomology, Agricultural Research & Extension Center, Texas A&M
University, 10345 Agnes Street, Corpus Christi, TX 78406, USA.
c-chilcutt@tamu.edu

We examined nine pairs of near-isogenic hybrids of Bacillus thuringiensis (Bt)
and non-Bt corn, Zea mays L., at two locations in 1999 and three locations in
2000 to compare the effects of Bt toxins on damage caused by Helicoverpa zea
(Boddie) to whorl stage field corn, and ear damage at harvest, as well as yield.
We found that whorl damage was less in all Bt hybrids compared with their non-Bt
counterparts each year and at each location. Differences in ear damage between Bt
and non-Bt hybrids, however, differed in 1999 and 2000. In 1999, only one Bt
hybrid, NC+5788Bt, had less ear damage than its non-Bt counterpart at the dryland
site, whereas four Bt hybrids, C8120Bt, P31B13Bt, P33VO8Bt, and NC+5788Bt, had
less damage at the irrigated site. In 2000, most Bt hybrids had less ear damage
than their non-Bt counterparts at each location. Differences in whorl damage did
not translate into yield differences. However, variations in ear damage were
partially reflected in yield differences. In 1999, P31B13Bt and P33V08Bt had
higher yields than their non-Bt counterparts at both sites, whereas in 2000 all
Bt hybrids had higher yields. Also, although whorl damage was not correlated with
yield, ear damage was negatively correlated with yield; increasing ear damage by
H. zea decreased yield for Bt and non-Bt hybrids alike. Overall, depending on
location and year, each centimeter of H. zea ear damage reduced yield by between
2 and 13%.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17195689 [PubMed - indexed for MEDLINE]

891: J Econ Entomol. 2006 Dec;99(6):2100-9.

Evaluation of oviposition deterrence in management of resistance to transgenic
corn by European corn borer (Lepidoptera: Crambidae).

Onstad DW, Buschman LL.

Department of Natural Resources and Environmental Science, University of
Illinois, Urbana, IL 61801, USA. onstad@uiuc.edu

We modified an existing model for European corn borer, Ostrinia nubilalis
(Hö¼bner) (Lepidoptera: Crambidae), population dynamics and genetics to evaluate
the effectiveness of oviposition deterrence in transgenic fields for resistance
management. We simulated two types of deterrence: one type has females reducing
their oviposition because of lost opportunities to lay eggs (eggs lost), and the
other type has the deterred females moving to the refuge to lay eggs. Oviposition
deterrence was clearly effective in extending the time to resistance to
transgenic corn (R allele) in the European corn borer, particularly when 80% or
more of the eggs were deterred from being oviposited on the transgenic plants.
With 90% of eggs deterred, the time required to reach 50% R-allele frequency
increases 3.7- to 5.5-fold compared with the no-deterrence scenario. The time to
50% R-allele frequency was similar for the two types of simulated deterrence, but
the densities of the European corn borer were 100-fold higher when the deterred
females oviposited in the refuge. The Y allele for insensitivity or resistance to
deterrence never reached 50% within the 50-yr time line for these simulations
except when the R allele was dominant and the Y allele was not recessive. The
time to 50% Y-allele frequency was 33 and 26 yr when the Y allele was additive or
dominant, respectively, when 50% of the eggs were deterred, but the time
decreased to 18 and 16 yr when 90% of the eggs were deterred. The effectiveness
of oviposition deterrence on time to resistance to transgenic insecticidal plants
was not changed much when we altered our assumptions about behavior in a
sensitivity analysis.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17195679 [PubMed - indexed for MEDLINE]

892: Plant Cell. 2006 Dec;18(12):3370-85. Epub 2006 Dec 22.

Defects in CTP:PHOSPHORYLETHANOLAMINE CYTIDYLYLTRANSFERASE affect embryonic and
postembryonic development in Arabidopsis.

Mizoi J, Nakamura M, Nishida I.

Department of Biological Sciences, Graduate School of Science, University of
Tokyo, Tokyo 113-0033, Japan.

A TILLING strategy (for targeting-induced local-scale lesions in genomes) was
used in Arabidopsis thaliana to isolate mutants of a gene encoding
CTP:PHOSPHORYLETHANOLAMINE CYTIDYLYLTRANSFERASE (PECT; EC 2.7.7.14), a
rate-limiting enzyme in phosphatidylethanolamine biosynthesis. A null mutation,
pect1-6, caused embryo abortion before the octant stage. However, reciprocal
crosses revealed that pect1-6 caused no significant gametophytic defect. In
pect1-4, PECT activity was decreased by 74%. Growth was generally normal in these
mutants, despite delays in embryo maturation and reduced fertility. At low
temperatures, however, homozygotic pect1-4 plants displayed dwarfism. PECT
activity was decreased by 47% in heterozygotic pect1-6 plants and by 80% in
pect1-4/pect1-6 F1 plants, which also displayed a small but significant decrease
of phosphatidylethanolamine and a reciprocal increase in phosphatidylcholine.
These lipid changes were fully reversed by wild-type PECT1 expression.
pect1-4/pect1-6 F1 plants displayed severe dwarfism, tissue abnormalities, and
low fertility, which was attributable in part to inhibition of anther, embryo,
and ovule development, as was the reduced fertility of pect1-4 seedlings. PECT1
cDNA expression under the control of an inducible promoter partially rectified
the mutant phenotypes observed in pect1-4/pect1-6 F1 seedlings, indicating that
malfunctions in different tissues have a synergistic effect on the mutant
phenotypes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17189343 [PubMed - indexed for MEDLINE]

893: Plant Cell. 2006 Dec;18(12):3443-57. Epub 2006 Dec 22.

Dynamics of a mobile RNA of potato involved in a long-distance signaling pathway.

Banerjee AK, Chatterjee M, Yu Y, Suh SG, Miller WA, Hannapel DJ.

Department of Horticulture, Iowa State University, Ames, Iowa 50011-1100, USA.

BEL1-like transcription factors interact with Knotted1 types to regulate numerous
developmental processes. In potato (Solanum tuberosum), the BEL1 transcription
factor St BEL5 and its protein partner POTH1 regulate tuber formation by
mediating hormone levels in the stolon tip. The accumulation of St BEL5 RNA
increases in response to short-day photoperiods, inductive for tuber formation.
RNA detection methods and heterografting experiments demonstrate that BEL5
transcripts are present in phloem cells and move across a graft union to localize
in stolon tips, the site of tuber induction. This movement of RNA to stolon tips
is correlated with enhanced tuber production. Overexpression of BEL5 transcripts
that include the untranslated sequences of the BEL5 transcript endows transgenic
lines with the capacity to overcome the inhibitory effects of long days on tuber
formation. Addition of the untranslated regions leads to preferential
accumulation of the BEL5 RNA in stolon tips under short-day conditions. Using a
leaf-specific promoter, the movement of BEL5 RNA to stolon tips was facilitated
by a short-day photoperiod, and this movement was correlated with enhanced tuber
production. These results implicate the transcripts of St BEL5 in a long-distance
signaling pathway that are delivered to the target organ via the phloem stream.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17189340 [PubMed - indexed for MEDLINE]

894: Plant Physiol. 2007 Feb;143(2):745-58. Epub 2006 Dec 22.

Ectopic expression of ABSCISIC ACID 2/GLUCOSE INSENSITIVE 1 in Arabidopsis
promotes seed dormancy and stress tolerance.

Lin PC, Hwang SG, Endo A, Okamoto M, Koshiba T, Cheng WH.

Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan,
Republic of China.

Abscisic acid (ABA) is an important phytohormone that plays a critical role in
seed development, dormancy, and stress tolerance. 9-cis-Epoxycarotenoid
dioxygenase is the key enzyme controlling ABA biosynthesis and stress tolerance.
In this study, we investigated the effect of ectopic expression of another ABA
biosynthesis gene, ABA2 (or GLUCOSE INSENSITIVE 1 [GIN1]) encoding a short-chain
dehydrogenase/reductase in Arabidopsis (Arabidopsis thaliana). We show that
ABA2-overexpressing transgenic plants with elevated ABA levels exhibited seed
germination delay and more tolerance to salinity than wild type when grown on
agar plates and/or in soil. However, the germination delay was abolished in
transgenic plants showing ABA levels over 2-fold higher than that of wild type
grown on 250 mm NaCl. The data suggest that there are distinct mechanisms
underlying ABA-mediated inhibition of seed germination under diverse stress. The
ABA-deficient mutant aba2, with a shorter primary root, can be restored to normal
root growth by exogenous application of ABA, whereas transgenic plants
overexpressing ABA2 showed normal root growth. The data reflect that the basal
levels of ABA are essential for maintaining normal primary root elongation.
Furthermore, analysis of ABA2 promoter activity with ABA2::beta-glucuronidase
transgenic plants revealed that the promoter activity was enhanced by multiple
prolonged stresses, such as drought, salinity, cold, and flooding, but not by
short-term stress treatments. Coincidently, prolonged drought stress treatment
led to the up-regulation of ABA biosynthetic and sugar-related genes. Thus, the
data support ABA2 as a late expression gene that might have a fine-tuning
function in mediating ABA biosynthesis through primary metabolic changes in
response to stress.

PMID: 17189333 [PubMed - indexed for MEDLINE]

895: Plant Physiol. 2007 Feb;143(2):639-49. Epub 2006 Dec 22.

Transgenic tobacco plants overexpressing chloroplastic ferredoxin-NADP(H)
reductase display normal rates of photosynthesis and increased tolerance to
oxidative stress.

Rodriguez RE, Lodeyro A, Poli HO, Zurbriggen M, Peisker M, Palatnik JF, Tognetti
VB, Tschiersch H, Hajirezaei MR, Valle EM, Carrillo N.

Instituto de Biologö­a Molecular y Celular de Rosario, Divisiö¨n Biologö­a
Molecular, Facultad de Ciencias Bioquö­micas y Farmacö©uticas, Universidad Nacional
de Rosario, S2002LRK Rosario, Argentina.

Ferredoxin-NADP(H) reductase (FNR) catalyzes the last step of photosynthetic
electron transport in chloroplasts, driving electrons from reduced ferredoxin to
NADP+. This reaction is rate limiting for photosynthesis under a wide range of
illumination conditions, as revealed by analysis of plants transformed with an
antisense version of the FNR gene. To investigate whether accumulation of this
flavoprotein over wild-type levels could improve photosynthetic efficiency and
growth, we generated transgenic tobacco (Nicotiana tabacum) plants expressing a
pea (Pisum sativum) FNR targeted to chloroplasts. The alien product distributed
between the thylakoid membranes and the chloroplast stroma. Transformants grown
at 150 or 700 micromol quanta m(-2) s(-1) displayed wild-type phenotypes
regardless of FNR content. Thylakoids isolated from plants with a 5-fold FNR
increase over the wild type displayed only moderate stimulation (approximately
20%) in the rates of electron transport from water to NADP+. In contrast, when
donors of photosystem I were used to drive NADP+ photoreduction, the activity was
3- to 4-fold higher than the wild-type controls. Plants expressing various levels
of FNR (from 1- to 3.6-fold over the wild type) failed to show significant
differences in CO2 assimilation rates when assayed over a range of light
intensities and CO2 concentrations. Transgenic lines exhibited enhanced tolerance
to photooxidative damage and redox-cycling herbicides that propagate reactive
oxygen species. The results suggest that photosynthetic electron transport has
several rate-limiting steps, with FNR catalyzing just one of them.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17189326 [PubMed - indexed for MEDLINE]

896: J Biosci Bioeng. 2006 Nov;102(5):375-89.

Developments in biotechnological production of sweet proteins.

Masuda T, Kitabatake N.

Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto
University, Uji, Kyoto 611-0011, Japan.

Most proteins are tasteless and flavorless, while some proteins elicit a
sweet-taste response on the human palate. Six proteins, thaumatin, monellin,
mabinlin, brazzein, egg lysozyme, and neoculin (previously considered as
curculin) have been identified as sweet-tasting proteins. However, no common
features among them have been observed. Herein, recent advances in the research
of sweet-tasting proteins and the production of such proteins by biotechnological
approaches are reviewed. Information on the structure-sweetness relationship for
these proteins would help not only in the clarification of the mechanism of
interaction of sweet-tasting proteins with their receptors, but also in the
design of more effective low-calorie sweeteners.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17189164 [PubMed - indexed for MEDLINE]

897: Vaccine. 2007 Feb 19;25(9):1647-57. Epub 2006 Nov 17.

Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed
and analysis of its immunogenicity as an oral vaccine.

Moravec T, Schmidt MA, Herman EM, Woodford-Thomas T.

Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, MO 63132,
United States.

The B subunit of the heat labile toxin of enterotoxigenic Escherichia coli (LTB)
was used as a model immunogen for production in soybean seed. LTB expression was
directed to the endoplasmic reticulum (ER) of seed storage parenchyma cells for
sequestration in de novo synthesized inert protein accretions derived from the
ER. Pentameric LTB accumulated to 2.4% of the total seed protein at maturity and
was stable in desiccated seed. LTB-soybean extracts administered orally to mice
induced both systemic IgG and IgA, and mucosal IgA antibody responses, and was
particularly efficacious when used in a parenteral prime-oral gavage boost
immunization strategy. Sera from immunized mice blocked ligand binding in vitro
and immunized mice exhibited partial protection against LT challenge. Moreover,
soybean-expressed LTB stimulated the antibody response against a co-administered
antigen by 500-fold. These results demonstrate the utility of soybean as an
efficient production platform for vaccines that can be used for oral delivery.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17188785 [PubMed - indexed for MEDLINE]

898: Int J Biol Macromol. 2007 Apr 10;40(5):449-60. Epub 2006 Nov 15.

Structural and thermodynamic properties of starches extracted from GBSS and GWD
suppressed potato lines.

Kozlov SS, Blennow A, Krivandin AV, Yuryev VP.

Institute of Biochemical Physics, Russian Academy of Sciences, Kosygina str. 4,
119334 Moscow, Russia.

A combined DSC-SAXS approach was employed to study the effects of amylose and
phosphate esters on the assembly structures of amylopectin in B-type polymorphic
potato tuber starches. Amylose and phosphate levels in the starches were
specifically engineered by antisense suppression of the granule bound starch
synthase (GBSS) and the glucan water dikinase (GWD), respectively. Joint analysis
of the SAXS and DSC data for the engineered starches revealed that the sizes of
amylopectin clusters, thickness of crystalline lamellae and the polymorphous
structure type remained unchanged. However, differences were found in the
structural organization of amylopectin clusters reflected in localization of
amylose within these supramolecular structures. Additionally, data for annealed
starches shows that investigated potato starches possess different types of
amylopectin defects. The relationship between structure of investigated potato
starches and their thermodynamic properties was recognized.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17188347 [PubMed - indexed for MEDLINE]

899: Science. 2007 Feb 23;315(5815):1098-103. Epub 2006 Dec 21.

Comment in:
Science. 2007 Feb 23;315(5815):1088-9.

Nuclear activity of MLA immune receptors links isolate-specific and basal
disease-resistance responses.

Shen QH, Saijo Y, Mauch S, Biskup C, Bieri S, Keller B, Seki H, Ulker B, Somssich
IE, Schulze-Lefert P.

Department of Plant Microbe Interactions, Max-Planck-Institut fö¼r
Zö¼chtungsforschung, Carl-von-Linnö©-Weg 10, D-50829 Kö¶ln, Germany.

Plant immune responses are triggered by pattern recognition receptors that detect
conserved pathogen-associated molecular patterns (PAMPs) or by resistance (R)
proteins recognizing isolate-specific pathogen effectors. We show that in barley,
intracellular mildew A (MLA) R proteins function in the nucleus to confer
resistance against the powdery mildew fungus. Recognition of the fungal
avirulence A10 effector by MLA10 induces nuclear associations between receptor
and WRKY transcription factors. The identified WRKY proteins act as repressors of
PAMP-triggered basal defense. MLA appears to interfere with the WRKY repressor
function, thereby de-repressing PAMP-triggered basal defense. Our findings reveal
a mechanism by which these polymorphic immune receptors integrate distinct
pathogen signals.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17185563 [PubMed - indexed for MEDLINE]

900: Risk Anal. 2006 Dec;26(6):1707-19.

Exploring the structure of attitudes toward genetically modified food.

Poortinga W, Pidgeon NF.

Cardiff University, Welsh School of Architecture, Cardiff, Wales, UK.
PoortingaW@Cardiff.ac.uk

Although it is often thought that the British public is opposed to genetically
modified (GM) food, recent qualitative work suggests that most people are
ambivalent about GM food and crops. In this article we explore the structure of
attitudes in order to examine whether attitudinal ambivalence can be captured by
more quantitative methods. Based on the finding that the perceived risks and
benefits of GM food can be treated as independent dimensions, we propose a
four-way typology of attitudes, consisting of a positive, negative, indifferent,
and ambivalent group. This study showed that the differences between the four
groups could best be described by three main dimensions: (1) a general evaluative
dimension, (2) an involvement dimension, and (3) an attitudinal certainty
dimension. While these different attitudinal dimensions have generally been
studied in isolation, we argue that they should be studied collectively.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17184407 [PubMed - indexed for MEDLINE]

901: Expert Opin Drug Deliv. 2007 Jan;4(1):1-3.

Transgenic probiotica as drug delivery systems: the golden bullet?

Yuvaraj S, Peppelenbosch MP, Bos NA.

Functional human proteins are constitutively produced in genetically modified
bacteria that survive on human mucosal surfaces, to the benefit of the host. The
successful Phase I clinical trial with IL-10-producing Lactococcus lactis for
Crohn's disease has opened new avenues for the use of transgenic bacteria as
delivery vehicles. The major advantage of this novel strategy is the avoidance of
systemic side effects associated with conventional therapies. This methodology
opens up an alternative method for local delivery of therapeutic proteins to
various mucosal tissues.

Publication Types:
Editorial

PMID: 17184157 [PubMed - indexed for MEDLINE]

902: Genet Mol Res. 2006 Nov 30;5(4):688-95.

Cassava in South America, Brazil's contribution and the lesson to be learned from
India.

Nassar NM.

Departamento de Genö©tica e Morfologia, Universidade de Brasö­lia, Brasö­lia, DF,
Brasil. nagnassa@rudah.com.br

South America is responsible for about half of the cassava world production. In
the 1970's productivity of the crop on the continent was about 15 ton/ha, and
dropped continuously until reaching 12 ton/ha in 2004. India's productivity of
cassava increased from 10 ton/ha in the 1970's to 28 ton/ha in 2004. Brazil
contributed significantly to improving cassava crops through the Instituto
Agronö´mico de Campinas in the 1960's and 1970's. The Universidade de Brasö­lia
released high-protein content hybrids, apomictic clones and explored the
potential of indigenous landraces.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17183479 [PubMed - indexed for MEDLINE]

903: Br J Nutr. 2006 Dec;96(6):997-1005.

Conventional and real-time polymerase chain reaction assessment of the fate of
transgenic DNA in sheep fed Roundup Ready rapeseed meal.

Alexander TW, Reuter T, Okine E, Sharma R, McAllister TA.

Agriculture and Agri-Food Canada Research Centre, Lethbridge, Alberta, Canada.

Conventional and real-time PCR were used to detect transgenic DNA in digesta,
faeces and blood collected from six ruminally and duodenally cannulated sheep fed
forage-based (F) or concentrate-based (C) diets containing 15% Roundup Ready (RR)
rapeseed meal (n 3). The sheep were adapted for 14 d to F or C diets containing
non-GM rapeseed, then fed the RR diets for 11 d. On day 12, they were switched
back to non-GM diets for a further 11 d. Ruminal and duodenal fluids (RF, DF) and
faecal samples were collected at 3 or 4 h intervals over the 4 d immediately
following the last feeding of GM diets. DNA was isolated from whole RF and DF,
from the cell-free supernatant fraction, and from culture fermentation liquid.
Blood was collected on days 1, 5 and 9 of feeding the RR rapeseed meal. The 1363
bp 5-enolpyruvylshikimate-3-phosphate synthase transgene (epsps) was quantifiable
in whole RF and DF for up to 13 h, and a 108 bp epsps fragment for up to 29 h.
Transgenic DNA was not detectable in faeces or blood, or in microbial DNA. Diet
type (F v. C) did not affect (P>0.05) the quantity of transgenic DNA in digesta.
More (P<0.05) transgenic DNA was detected in RF than in DF, but there was an
interaction (P<0.05) between sample type and collection time. In supernatant
fractions from RF and DF, three different fragments of transgenic DNA ranging in
size from 62 to 420 bp were not amplifiable.

PMID: 17181873 [PubMed - indexed for MEDLINE]

904: Plant J. 2007 Feb;49(3):414-27. Epub 2006 Dec 20.

Red colouration in apple fruit is due to the activity of the MYB transcription
factor, MdMYB10.

Espley RV, Hellens RP, Putterill J, Stevenson DE, Kutty-Amma S, Allan AC.

HortResearch, Mt Albert Research Centre, Private Bag 92169, Auckland, New
Zealand.

Anthocyanin concentration is an important determinant of the colour of many
fruits. In apple (Malus x domestica), centuries of breeding have produced
numerous varieties in which levels of anthocyanin pigment vary widely and change
in response to environmental and developmental stimuli. The apple fruit cortex is
usually colourless, although germplasm does exist where the cortex is highly
pigmented due to the accumulation of either anthocyanins or carotenoids. From
studies in a diverse array of plant species, it is apparent that anthocyanin
biosynthesis is controlled at the level of transcription. Here we report the
transcript levels of the anthocyanin biosynthetic genes in a red-fleshed apple
compared with a white-fleshed cultivar. We also describe an apple MYB
transcription factor, MdMYB10, that is similar in sequence to known anthocyanin
regulators in other species. We further show that this transcription factor can
induce anthocyanin accumulation in both heterologous and homologous systems,
generating pigmented patches in transient assays in tobacco leaves and highly
pigmented apple plants following stable transformation with constitutively
expressed MdMYB10. Efficient induction of anthocyanin biosynthesis in transient
assays by MdMYB10 was dependent on the co-expression of two distinct bHLH
proteins from apple, MdbHLH3 and MdbHLH33. The strong correlation between the
expression of MdMYB10 and apple anthocyanin levels during fruit development
suggests that this transcription factor is responsible for controlling
anthocyanin biosynthesis in apple fruit; in the red-fleshed cultivar and in the
skin of other varieties, there is an induction of MdMYB10 expression concurrent
with colour formation during development. Characterization of MdMYB10 has
implications for the development of new varieties through classical breeding or a
biotechnological approach.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17181777 [PubMed - indexed for MEDLINE]

905: Transgenic Res. 2007 Aug;16(4):531-8. Epub 2006 Dec 19.

Use of a PTGS-MAR expression system for efficient in planta production of
bioactive Arabidopsis thaliana plant defensins.

Sels J, Delaurö© SL, Aerts AM, Proost P, Cammue BP, De Bolle MF.

Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven,
Kasteelpark Arenberg 20, Heverlee, Belgium.

Plant defensins, exhibiting various levels of inhibitory activity against fungal
pathogens, are potent candidates for pharmaceutical or agricultural antimycotics.
Study of the plant defensins from the model plant Arabidopsis thaliana requires
the purification of these peptides. However, heterologous production of defensins
for large-scale in vitro bioactivity assays is often experienced as a major
problem. In this study we describe the transgenic expression of a previously
identified seed-specific and a so far uncharacterized plant defensin gene in
their host A. thaliana using a formerly developed plant expression system.
Therefore, both genes were cloned in a matrix attachment region (MAR) based plant
transformation vector and expressed in post-transcriptional gene silencing (PTGS)
impaired A. thaliana plants. The peptides were purified to homogeneity and were
correctly processed, as confirmed by mass spectrometry analysis. Finally, they
were assessed for their in vitro antifungal activity and mode of antifungal
action. Our results indicate that the PTGS-MAR expression system can be applied
to obtain significant amounts of bioactive, rightly processed plant peptides from
leaves of first generation transgenic plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17180735 [PubMed - indexed for MEDLINE]

906: Biochem Biophys Res Commun. 2007 Feb 9;353(2):299-305. Epub 2006 Dec 13.

GmDREB2, a soybean DRE-binding transcription factor, conferred drought and
high-salt tolerance in transgenic plants.

Chen M, Wang QY, Cheng XG, Xu ZS, Li LC, Ye XG, Xia LQ, Ma YZ.

National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI),
Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, Institute
of Crop Science, Chinese Academy of Agricultural Sciences (CAAS), Beijing 100081,
China.

A novel DREB (dehydration responsive element binding protein) homologous gene,
GmDREB2, was isolated from soybean. Based on its similarity with AP2 domains,
GmDREB2 was classified into A-5 subgroup in DREB subfamily in AP2/EREBP family.
Expression of GmDREB2 gene was induced by drought, high salt, and low temperature
stresses and abscisic acid treatment. The GmDREB2 bound specifically to DRE
element in vitro. Furthermore, the overexpression of GmDREB2 activated expression
of downstream genes in transgenic Arabidopsis, resulting in enhanced tolerance to
drought and high-salt stresses and did not cause growth retardation. Analysis of
free proline contents in transgenic tobacco indicated that the overexpression of
GmDREB2 accumulated higher level of free proline compared to the wild type plants
under drought condition. The results from this study indicate that this novel
soybean GmDREB2 gene functions as an important transcriptional activator and may
be useful in improving of plant tolerance to abiotic stresses in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17178106 [PubMed - indexed for MEDLINE]

907: Plant Biotechnol J. 2006 Jul;4(4):467-75.

Marker-free transgenic (MFT) near-isogenic introgression lines (NIILs) of
'golden' indica rice (cv. IR64) with accumulation of provitamin A in the
endosperm tissue.

Baisakh N, Rehana S, Rai M, Oliva N, Tan J, Mackill DJ, Khush GS, Datta K, Datta
SK.

Plant Breeding, Genetics, and Biotechnology Division, International Rice Research
Institute, DAPO Box 7777, Metro Manila, Philippines. nbaisakh@agctr.lsu.edu

We have developed near-isogenic introgression lines (NIILs) of an elite indica
rice cultivar (IR64) with the genes for beta-carotene biosynthesis from dihaploid
(DH) derivatives of golden japonica rice (cv. T309). A careful analysis of the DH
lines indicated the integration of the genes of interest [phytoene synthase (psy)
and phytoene desaturase (crtI)] and the selectable marker gene (hygromycin
phosphotransferase, hph) in two unlinked loci. During subsequent crossing,
progenies could be obtained carrying only the locus with psy and crtI, which was
segregated independently from the locus containing the hph gene during meiotic
segregation. The NIILs (BC(2)F(2)) showed maximum similarity with the recurrent
parent cultivar IR64. Further, progenies of two NIILs were devoid of any
fragments beyond the left or right border, including the chloramphenicol
acetyltransferase (cat) antibiotic resistance gene of the transformation vector.
Spectrophotometric readings showed the accumulation of up to 1.06 microg total
carotenoids, including beta-carotene, in 1 g of the endosperm. The accumulation
of beta-carotene was also evident from the clearly visible yellow colour of the
polished seeds.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17177811 [PubMed - indexed for MEDLINE]

908: Plant Biotechnol J. 2006 Jul;4(4):433-44.

Pathway engineering for healthy phytochemicals leading to the production of novel
flavonoids in tomato fruit.

Schijlen E, Ric de Vos CH, Jonker H, van den Broeck H, Molthoff J, van Tunen A,
Martens S, Bovy A.

Plant Research International, Business Unit Bioscience, PO Box 16, 6700 AA
Wageningen, The Netherlands. elio.schijlen@wur.nl

Flavonoids are a large family of plant polyphenolic secondary metabolites.
Although they are widespread throughout the plant kingdom, some flavonoid classes
are specific for only a few plant species. Due to their presumed health benefits
there is growing interest in the development of food crops with tailor-made
levels and composition of flavonoids, designed to exert an optimal biological
effect. In order to explore the possibilities of flavonoid engineering in tomato
fruits, we have targeted this pathway towards classes of potentially healthy
flavonoids which are novel for tomato. Using structural flavonoid genes (encoding
stilbene synthase, chalcone synthase, chalcone reductase, chalcone isomerase and
flavone synthase) from different plant sources, we were able to produce
transgenic tomatoes accumulating new phytochemicals. Biochemical analysis showed
that the fruit peel contained high levels of stilbenes (resveratrol and piceid),
deoxychalcones (butein and isoliquiritigenin), flavones (luteolin-7-glucoside and
luteolin aglycon) and flavonols (quercetin glycosides and kaempferol glycosides).
Using an online high-performance liquid chromatography (HPLC) antioxidant
detection system, we demonstrated that, due to the presence of the novel
flavonoids, the transgenic tomato fruits displayed altered antioxidant profiles.
In addition, total antioxidant capacity of tomato fruit peel with high levels of
flavones and flavonols increased more than threefold. These results on genetic
engineering of flavonoids in tomato fruit demonstrate the possibilities to change
the levels and composition of health-related polyphenols in a crop plant and
provide more insight in the genetic and biochemical regulation of the flavonoid
pathway within this worldwide important vegetable.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17177808 [PubMed - indexed for MEDLINE]

909: Plant Biotechnol J. 2006 Jul;4(4):419-32.

Tomato is a highly effective vehicle for expression and oral immunization with
Norwalk virus capsid protein.

Zhang X, Buehner NA, Hutson AM, Estes MK, Mason HS.

Department of Plant Biology, Cornell University, Ithaca, NY 18853-1801, USA.
zhangxi@rockefeller.edu

Norwalk virus (NV) is an important agent of epidemic gastroenteritis, and an oral
subunit vaccine shows potential for protection. Recombinant Norwalk virus (rNV)
capsid protein expressed in plants assembles virus-like particles (VLPs) that are
orally immunogenic in mice and humans. In this article we examine rNV expression
in tomato and potato using a plant-optimized gene, and test the immunogenicity of
dried tomato fruit and potato tuber fed to mice. The synthetic gene increased rNV
expression fourfold in tomato and potato plants, which assembled VLP. Four doses
of 0.4 g freeze-dried tomato fruit containing 64 microg rNV (40 microg VLPs)
induced NV-specific serum IgG and mucosal IgA in > or = 80% of mice, while doses
of 0.8 g elicited systemic and mucosal antibody responses in all mice. Feedings
of 1 g freeze-dried potato tuber containing 120 microg rNV (90 microg VLPs) were
required to produce 100% responsiveness. Oxidation of phenolic compounds upon
rehydration of dried tuber caused significant VLP instability, thus decreasing
immunogenicity. Air-dried tomato fruit stimulated stronger immune responses than
freeze-dried fruit of the same mass, perhaps by limiting the destruction of plant
cell matrix and membrane systems that occurs with freeze-drying. Thus, rNV in
dried transgenic tomato fruit was a more potent immunogen than that in dried
potato tubers, based on the total VLPs ingested. These findings support the use
of stabilized, dried tomato fruit for oral delivery of subunit vaccines.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17177807 [PubMed - indexed for MEDLINE]

910: Plant Biotechnol J. 2006 Jul;4(4):409-18.

Production of high-starch, low-glucose potatoes through over-expression of the
metabolic regulator SnRK1.

McKibbin RS, Muttucumaru N, Paul MJ, Powers SJ, Burrell MM, Coates S, Purcell PC,
Tiessen A, Geigenberger P, Halford NG.

Long Ashton Research Station, Bristol BS41 9AF, UK.

Transgenic potato (Solanum tuberosum cv. Prairie) lines were produced
over-expressing a sucrose non-fermenting-1-related protein kinase-1 gene (SnRK1)
under the control of a patatin (tuber-specific) promoter. SnRK1 activity in the
tubers of three independent transgenic lines was increased by 55%-167% compared
with that in the wild-type. Glucose levels were decreased, at 17%-56% of the
levels of the wild-type, and the starch content showed an increase of 23%-30%.
Sucrose and fructose levels in the tubers of the transgenic plants did not show a
significant change. Northern analyses of genes encoding sucrose synthase and
ADP-glucose pyrophosphorylase, two key enzymes involved in the biosynthetic
pathway from sucrose to starch, showed that the expression of both was increased
in tubers of the transgenic lines compared with the wild-type. In contrast, the
expression of genes encoding two other enzymes of carbohydrate metabolism,
alpha-amylase and sucrose phosphate synthase, showed no change. The activity of
sucrose synthase and ADP-glucose pyrophosphorylase was also increased, by
approximately 20%-60% and three- to five-fold, respectively, whereas the activity
of hexokinase was unchanged. The results are consistent with a role for SnRK1 in
regulating carbon flux through the storage pathway to starch biosynthesis. They
emphasize the importance of SnRK1 in the regulation of carbohydrate metabolism
and resource partitioning, and indicate a specific role for SnRK1 in the control
of starch accumulation in potato tubers.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17177806 [PubMed - indexed for MEDLINE]

911: Plant Biotechnol J. 2006 Jul;4(4):381-92.

A metabolomic study of substantial equivalence of field-grown genetically
modified wheat.

Baker JM, Hawkins ND, Ward JL, Lovegrove A, Napier JA, Shewry PR, Beale MH.

National Centre for Plant and Microbial Metabolomics, Crop Performance and
Improvement Division, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK.

The 'substantial equivalence' of three transgenic wheats expressing additional
high-molecular-weight subunit genes and the corresponding parental lines (two
lines plus a null transformant) was examined using metabolite profiling of
samples grown in replicate field trials on two UK sites (Rothamsted,
Hertfordshire and Long Ashton, near Bristol) for 3 years. Multivariate comparison
of the proton nuclear magnetic resonance spectra of polar metabolites extracted
with deuterated methanol-water showed a stronger influence of site and year than
of genotype. Nevertheless, some separation between the transgenic and parental
lines was observed, notably between the transgenic line B73-6-1 (which had the
highest level of transgene expression) and its parental line L88-6. Comparison of
the spectra showed that this separation resulted from increased levels of maltose
and/or sucrose in this transgenic line, and that differences in free amino acids
were also apparent. More detailed studies of the amino acid composition of
material grown in 2000 were carried out using gas chromatography-mass
spectrometry. The most noticeable difference was that the samples grown at
Rothamsted consistently contained larger amounts of acidic amino acids (glutamic,
aspartic) and their amides (glutamine, asparagine). In addition, the related
lines, L88-6 and B73-6-1, both contained larger amounts of proline and
gamma-aminobutyric acid when grown at Long Ashton than at Rothamsted. The results
clearly demonstrate that the environment affects the metabolome and that any
differences between the control and transgenic lines are generally within the
same range as the differences observed between the control lines grown on
different sites and in different years.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17177804 [PubMed - indexed for MEDLINE]

912: Plant Biotechnol J. 2006 Jul;4(4):369-80.

Transgenesis has less impact on the transcriptome of wheat grain than
conventional breeding.

Baudo MM, Lyons R, Powers S, Pastori GM, Edwards KJ, Holdsworth MJ, Shewry PR.

Rothamsted Research, Harpenden AL5 2JQ, UK.

Detailed global gene expression profiles have been obtained for a series of
transgenic and conventionally bred wheat lines expressing additional genes
encoding HMW (high molecular weight) subunits of glutenin, a group of
endosperm-specific seed storage proteins known to determine dough strength and
therefore bread-making quality. Differences in endosperm and leaf transcriptome
profiles between untransformed and derived transgenic lines were consistently
extremely small, when analysing plants containing either transgenes only, or also
marker genes. Differences observed in gene expression in the endosperm between
conventionally bred material were much larger in comparison to differences
between transgenic and untransformed lines exhibiting the same complements of
gluten subunits. These results suggest that the presence of the transgenes did
not significantly alter gene expression and that, at this level of investigation,
transgenic plants could be considered substantially equivalent to untransformed
parental lines.

Publication Types:
Comparative Study
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17177803 [PubMed - indexed for MEDLINE]

913: J Agric Food Chem. 2006 Dec 27;54(26):9901-5.

Biochemical safety evaluation of transgenic rice seeds expressing T cell epitopes
of Japanese cedar pollen allergens.

Takagi H, Hirose S, Yasuda H, Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan.

Transgenic rice seeds, which express a hybrid peptide comprising seven
predominant human T cell epitopes (7Crp) derived from Japanese cedar pollen
allergens, have been shown to function as an effective edible vaccine for the
control of pollen allergen-induced responses. In this study, we characterized
biochemical properties of transgenic seeds expressing the 7Crp peptide. The
levels of chemical compositions, such as carbohydrate, protein, lipid, amino
acid, fatty acid, mineral, and vitamin, were substantially equivalent between
transgenic 7Crp and its nontransgenic counterpart seeds. The contents of three
major allergenic proteins in transgenic seeds were not enhanced by expression of
the 7Crp peptide when compared with those of nontransgenic seeds. The 7Crp
peptide expressed in seeds was susceptible to simulated gastric/intestinal
fluids. N-Glycosylation was not observed in the 7Crp peptide sequence. These
results indicate that transgenic 7Crp seeds are substantially equivalent to
nontransgenic parental seeds except for the presence of the 7Crp peptide.
Keywords: Food safety assessment; transgenic rice seed; edible vaccine;
peptide-based immunotherapy; Japanese cedar pollinosis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17177518 [PubMed - indexed for MEDLINE]

914: J Agric Food Chem. 2006 Dec 27;54(26):9882-7.

Improving potato storage and processing characteristics through all-native DNA
transformation.

Rommens CM, Ye J, Richael C, Swords K.

J. R. Simplot Company, Simplot Plant Sciences, Boise, Idaho 83706, USA.

The dominant potato (Solanum tuberosum) variety for French fry production in the
United States is the 131-year-old Russet Burbank. Market penetration of the
higher yielding and more uniform Ranger Russet variety is limited to about
one-fifth of that of the Russet Burbank because of two storage deficits: black
spot bruise sensitivity and high levels of cold-induced sweetening. Here, these
trait weaknesses are turned into strengths by simultaneously lowering the
expression of Ranger Russet's tuber-expressed polyphenol oxidase (Ppo),
starch-associated R1, and phosphorylase-L (PhL) genes. This genetic modification
was accomplished without inserting any foreign DNA into the plant genome. French
fries from the intragenic potatoes also contained reduced amounts of the
antinutritional compound acrylamide while, unexpectedly, displaying enhanced
sensory characteristics.

PMID: 17177515 [PubMed - indexed for MEDLINE]

915: J Agric Food Chem. 2006 Dec 27;54(26):9658-63.

Equal performance of TaqMan, MGB, molecular beacon, and SYBR green-based
detection assays in detection and quantification of roundup ready soybean.

Andersen CB, Holst-Jensen A, Berdal KG, Thorstensen T, Tengs T.

Section of Food and Feed Microbiology, National Veterinary Institute,
Ullevaalsveien 68, 0454 Oslo, Norway.

We have tested and compared the performance of 12 different assays representing
four different real-time polymerase chain reaction (PCR) chemistries in the
context of genetically modified organism detection. Several different molecular
beacon, SYBR Green, TaqMan, and MGB assays were designed for the event specific
detection and quantification of the 3' integration junction of GTS 40-3-2
(Roundup Ready) soybean. Sensitivity as well as robustness in the presence of
background DNA were tested. None of the PCR-based approaches appeared to be
significantly better than any of the other, but the molecular beacon assays had
the lowest efficiency and also seemed more sensitive to changes in experimental
setup.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17177484 [PubMed - indexed for MEDLINE]

916: Biochemistry. 2006 Dec 26;45(51):15179-87. Epub 2006 Dec 1.

Microsomal triglyceride transfer protein is required for yolk lipid utilization
and absorption of dietary lipids in zebrafish larvae.

Schlegel A, Stainier DY.

Department of Biochemistry and Biophysics, University of California, MC-2711,
1550 Fourth Street, Room 381, San Francisco, California 94143, USA.
amnon.schlegel@ucsf.edu

Although the absorption, transport, and catabolism of dietary lipids have been
studied extensively in great detail in mammals and other vertebrates, a tractable
genetic system for identifying novel genes involved in these physiologic
processes is not available. To establish such a model, we monitored neutral lipid
by staining fixed zebrafish larvae with oil red o (ORO). The head structures,
heart, vasculature, and swim bladder stained with ORO until the yolk was consumed
6 days after fertilization (6 dpf). Thereafter, the heart and vasculature no
longer had stainable neutral lipids. Following a high-fat meal, ORO stained the
intestine and vasculature of 6 dpf larvae, and whole-larval triacylglycerol (TAG)
and apolipoprotein B levels increased. Levels of microsomal triglyceride transfer
protein (Mtp), the protein responsible for packaging TAG and betalipoproteins
into lipoprotein particles, were unchanged by feeding. Since the developing
zebrafish embryo expresses mtp in the yolk cell layer, liver, and intestine, we
determined the effect of targeted knockdown of Mtp expression using an antisense
morpholino oligonucleotide approach (Mtp MO) on the transport of yolk and dietary
lipids. Mtp MO injection led to loss of Mtp expression and of lipid staining in
the vasculature, heart, and head structures. Mtp MO-injected larvae were smaller
than age-matched, uninjected larvae, consumed very little yolk, and did not
absorb dietary neutral lipids; however, they absorbed a short chain fatty acid
that does not require Mtp for transport. Importantly, the vasculature appeared
unaffected in Mtp MO-injected larvae. These studies indicate that zebrafish
larvae are suitable for genetic studies of lipid transport and metabolism.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17176039 [PubMed - indexed for MEDLINE]

917: Biofizika. 2006 Nov-Dec;51(6):1033-43.

[Population dynamics: limits of predictability]

[Article in Russian]

Medvinskiä­ AB.

Problems pertaining to the complex character of ecological system dynamics are
discussed. Examples of the complex dynamics of plankton populations in a
heterogeneous environment and agricultural ecosystems under invasion of pests
resistant to Bt toxins produced by genetically modified insecticidal crops are
given.

Publication Types:
English Abstract
Review

PMID: 17175915 [PubMed - indexed for MEDLINE]

918: J Exp Bot. 2007;58(3):637-49. Epub 2006 Dec 14.

Altered cytokinin metabolism affects cytokinin, auxin, and abscisic acid contents
in leaves and chloroplasts, and chloroplast ultrastructure in transgenic tobacco.

Polanskö¡ L, Vicö¡nkovö¡ A, Novö¡kovö¡ M, Malbeck J, Dobrev PI, Brzobohaty B, Vankovö¡
R, Machö¡ckovö¡ I.

Institute of Experimental Botany AS CR, Rozvojovö¡ 135, 165 02 Prague 6-Lysolaje,
Czech Republic. polanska@ueb.cas.cz

Cytokinins (CKs) are involved in the regulation of plant development including
plastid differentiation and function. Partial location of CK biosynthetic
pathways in plastids suggests the importance of CKs for chloroplast development.
The impact of genetically modified CK metabolism on endogenous CK,
indole-3-acetic acid, and abscisic acid contents in leaves and isolated intact
chloroplasts of Nicotiana tabacum was determined by liquid chromatography/mass
spectrometry and two-dimensional high-performance liquid chromatography, and
alterations in chloroplast ultrastructure by electron microscopy. Ectopic
expression of Sho, a gene encoding a Petunia hybrida isopentenyltransferase, was
employed to raise CK levels. The increase in CK levels was lower in chloroplasts
than in leaves. CK levels were reduced in leaves of tobacco harbouring a CK
oxidase/dehydrogenase gene, AtCKX3. The total CK content also decreased in
chloroplasts, but CK phosphate levels were higher than in the wild type. In a
transformant overexpressing a maize beta-glucosidase gene, Zm-p60.1, naturally
targeted to plastids, a decrease of CK-O-glucosides in chloroplasts was found. In
leaves, the changes were not significant. CK-O-glucosides accumulated to very
high levels in leaves, but not in chloroplasts, of plants overexpressing a ZOG1
gene, encoding trans-zeatin-O-glucosyltransferase from Phaseolus lunatus.
Manipulation of the CK content affected levels of indole-3-acetic and abscisic
acid. Chloroplasts of plants constitutively overexpressing Sho displayed
ultrastructural alterations including the occasional occurrence of crystalloids
and an increased number of plastoglobuli. The other transformants did not exhibit
any major differences in chloroplast ultrastructure. The results suggest that
plant hormone compartmentation plays an important role in hormone homeostasis and
that chloroplasts are rather independent organelles with respect to regulation of
CK metabolism.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17175552 [PubMed - indexed for MEDLINE]

919: Insect Biochem Mol Biol. 2007 Jan;37(1):10-8. Epub 2006 Oct 4.

The cyanogenic glucoside composition of Zygaena filipendulae (Lepidoptera:
Zygaenidae) as effected by feeding on wild-type and transgenic lotus populations
with variable cyanogenic glucoside profiles.

Zagrobelny M, Bak S, Ekströ¸m CT, Olsen CE, Mö¸ller BL.

Department of Plant Biology and Center for Molecular Plant Physiology (PlaCe), 40
Thorvaldsensvej, DK-1871 Frederiksberg C, Copenhagen, Denmark. miz@kvl.dk

Zygaena larvae sequester the cyanogenic glucosides linamarin and lotaustralin
from their food plants (Fabaceae) as well as carry out de novo biosynthesis of
these compounds. In this study, Zygaena filipendulae were reared on wild-type
Lotus corniculatus and wild-type and transgenic L. japonicus plants with
differing content and ratios of the cyanogenic glucosides linamarin and
lotaustralin and of the cyanoalkenyl glucosides rhodiocyanoside A and D. LC-MS
analyses, free choice feeding experiments and developmental studies were used to
examine the effect of varying content and ratios of these secondary metabolites
on the feeding preferences, growth and development of Z. filipendulae. Larvae
reared on cyanogenic L. corniculatus developed faster compared to larvae reared
on L. japonicus although free choice feeding trials demonstrated that the latter
plant source was the preferred food plant. Larvae reared on acyanogenic L.
corniculatus showed decelerated development. Analysis of different life stages
and tissues demonstrate that Z. filipendulae strive to maintain certain threshold
content and ratios of cyanogenic glucosides regardless of the composition of the
food plants. Despite this, the ratios of cyanogenic glucosides in Z. filipendulae
remain partly affected by the ratio of the food plant due to the high proportion
of sequestering that takes place.

PMID: 17175442 [PubMed - indexed for MEDLINE]

920: Methods Mol Biol. 2007;354:161-71.

Use of RNA interference to dissect defense-signaling pathways in rice.

Mei C, Zhou X, Yang Y.

Department of Plant Pathology, University of Arkansas, Fayetteville, USA.

The RNA interference (RNAi) technique is a powerful tool to suppress gene
expression and has been widely used for functional discovery of eukaryotic genes.
To dissect defense-signaling pathways in rice, it is important to generate a
series of rice mutant lines deficient in or insensitive to major signal molecules
such as jasmonic acid and ethylene. Here we describe an RNAi protocol for
generating and characterizing transgenic gene-silencing lines defective in rice
jasmonic acid signaling. The RNAi technique should be useful for effective
suppression of host genes encoding signaling components and facilitating the
dissection of defense signal pathways in rice.

PMID: 17172753 [PubMed - indexed for MEDLINE]

921: Sci China C Life Sci. 2006 Oct;49(5):414-28.

Adapting rice anther culture to gene transformation and RNA interference.

Chen C, Xiao H, Zhang W, Wang A, Xia Z, Li X, Zhai W, Cheng Z, Zhu L.

State Key Laboratory of Plant Genomics and National Center for Plant Gene
Research, Institute of Genetics and Developmental Biology, Chinese Academy of
Sciences, Beijing 100101, China.

Anther culture offers a rapid method of generating homozygous lines for breeding
program and genetic analysis. To produce homozygous transgenic lines of rice
(Oryza sativa L.) in one step, we developed an efficient protocol of
anther-callus-based transformation mediated by Agrobacterium after optimizing
several factors influencing efficient transformation, including callus induction
and Agrobacterium density for co-cultivation. Using this protocol, we obtained
145 independent green transformants from five cultivars of japonica rice by
transformation with a binary vector pCXK1301 bearing the rice gene, Xa21 for
resistance to bacterial blight, of which 140 were further confirmed by PCR and
Southern hybridization analysis, including haploids (32.1%), diploids (62.1%) and
mixoploids (7.5%). Fifteen diploids were found to be doubled haploids, which
accounted for 10.7% of the total positive lines. Finally, by including 28 from
colchicine induced or spontaneous diploidization of haploids later after
transformation, a total of 43 doubled haploids (30.7%) of Xa21 transgenic lines
were obtained. We also generated two RNAi transgenic haploids of the rice OsMADS2
gene, a putative redundant gene of OsMADS4 based on their sequence similarity, to
investigate its possible roles in rice flower development by this method. Flowers
from the two OsMADS2 RNAi transgenic haploids displayed obvious homeotic
alternations, in which lodicules were transformed into palea/lemma-like tissues,
whereas identities of other floral organs were maintained. The phenotypic
alternations were proved to result from specific transcriptional suppression of
OsMADS2 gene by the introduced RNAi transgene. The results confirmed that OsMADS2
is involved in lodicule development of rice flower and functionally redundant
with OsMADS4 gene. Our results demonstrated that rice anther culture could be
adapted to gene transformation and RNAi analysis in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17172048 [PubMed - indexed for MEDLINE]

922: Ontogenez. 2006 Nov-Dec;37(6):449-56.

[Fasciation in pea: basic principles of morphogenesis]

[Article in Russian]

Siniushin AA, Gostimskiä­ SA.

A study of fasciated pea Pisum sativum L. (Fabaceae) mutant Shtambovy in
comparison with the wild type (Nemchinovsky cultivar) has shown that fasciation
is a result of abnormal cohesion of axial or other structures which arise in a
superfluous amount due to uncontrolled meristic processes. In some cases, the
organs with the same number and position as in the wild type can be fascinated.
Subsequent defasciation and some features of tissue differentiation suggest that
the meristem of a fasciated shoot retains a certain degree of discreteness which
reflects its complex structure. The number and position of leaves in a node is a
function of the diameter of the leaf primordium inhibitory zone, size of the
shoot apical meristem, and number of bundles in a shoot. In the absence of the
apex proliferative activity combined with the reduction of phyllomes in the upper
nodes, abnormal cohesion of the second order axes, racemes, can take place. As a
result, inflorescences of special type develop.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 17168381 [PubMed - indexed for MEDLINE]

923: Plant Cell Rep. 2007 May;26(5):661-71. Epub 2006 Dec 13.

The usefulness of the gfp reporter gene for monitoring Agrobacterium-mediated
transformation of potato dihaploid and tetraploid genotypes.

Rakosy-Tican E, Aurori CM, Dijkstra C, Thieme R, Aurori A, Davey MR.

Babes-Bolyai University, Plant Genetic Engineering Group, 400006 Cluj-Napoca,
Romania. lrakosy@hasdeu.ubbcluj.ro

Potato is one of the main targets for genetic improvement by gene transfer. The
aim of the present study was to establish a robust protocol for the genetic
transformation of three dihaploid and four economically important cultivars of
potato using Agrobacterium tumefaciens carrying the in vivo screenable reporter
gene for green fluorescent protein (gfp) and the marker gene for neomycin
phosphotransferase (nptII). Stem and leaf explants were used for transformation
by Agrobacterium tumefaciens strain LBA4404 carrying the binary vector pHB2892.
Kanamycin selection, visual screening of GFP by epifluorescent microscopy, PCR
amplification of nptII and gfp genes, as well as RT-PCR and Southern blotting of
gfp and Northern blotting of nptII, were used for transgenic plant selection,
identification and analysis. Genetic transformation was optimized for the best
performing genotypes with a mean number of shoots expressing gfp per explant of
13 and 2 (dihaploid line 178/10 and cv. 'Baltica', respectively). The nptII
marker and gfp reporter genes permitted selection and excellent visual screening
of transgenic tissues and plants. They also revealed the effects of antibiotic
selection on organogenesis and transformation frequency, and the identification
of escapes and chimeras in all potato genotypes. Silencing of the gfp transgene
that may represent site-specific inactivation during cell differentiation,
occurred in some transgenic shoots of tetraploid cultivars and in specific
chimeric clones of the dihaploid line 178/10. The regeneration of escapes could
be attributed to either the protection of non-transformed cells by neighbouring
transgenic cells, or the persistence of Agrobacterium cells in plant tissues
after co-cultivation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17165042 [PubMed - indexed for MEDLINE]

924: Plant J. 2007 Jan;49(1):91-102. Epub 2006 Dec 6.

The Rc and Rd genes are involved in proanthocyanidin synthesis in rice pericarp.

Furukawa T, Maekawa M, Oki T, Suda I, Iida S, Shimada H, Takamure I, Kadowaki K.

Genetic Diversity Department, National Institute of Agrobiological Sciences,
Tsukuba, Ibaraki, Japan.

Different colors, such as purple, brown, red and white, occur in the pericarp of
rice. Here, two genes affecting proanthocyanidin synthesis in red- and
brown-colored rice were elucidated. Genetic segregation analysis suggested that
the Rd and A loci are identical, and both encode dihydroflavonol-4-reductase
(DFR). The introduction of the DFR gene into an Rcrd mutant resulted in
red-colored rice, which was brown in the original mutant, demonstrating that the
Rd locus encodes the DFR protein. Accumulation of proanthocyanidins was observed
in the transformants by the introduction of the Rd gene into the rice Rcrd line.
Protein blot analysis showed that the DFR gene was translated in seeds with
alternative translation initiation. A search for the Rc gene, which encodes a
transacting regulatory factor, was conducted using available DNA markers and the
Rice Genome Automated Annotation System program. Three candidate genes were
identified and cloned from a rice RcRd line and subsequently introduced into a
rice rcrd line. Brown-colored seeds were obtained from transgenic plants by the
introduction of a gene containing the basic helix-loop-helix (bHLH) motif,
demonstrating that the Rc gene encodes a bHLH protein. Comparison of the Rc locus
among rice accessions showed that a 14-bp deletion occurred only in the rc locus.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17163879 [PubMed - indexed for MEDLINE]

925: Cell Mol Biol Lett. 2007;12(2):206-19. Epub 2006 Dec 12.

Diploid potato (Solanum tuberosum L.) as a model crop to study transgene
expression.

Nadolska-Orczyk A, Pietrusinska A, Binka-Wyrwa A, Kuc D, Orczyk W.

Plant Transformation and Cell Engineering Department, Plant Breeding and
Acclimatization Institute, Radzikö¨w, Bå‚onie, Poland. a.orczyk@ihar.edu.pl

This paper presents a method of Agrobacterium-mediated transformation for two
diploid breeding lines of potato, and gives a detailed analysis of reporter gene
expression. In our lab, these lines were also used to obtain tetraploid somatic
hybrids. We tested four newly prepared constructs based on the pGreen vector
system containing the selection gene nptII or bar under the 35S or nos promoter.
All these vectors carried gus under 35S. We also tested the pDM805 vector, with
the bar and gus genes respectively under the Ubi1 and Act1 promoters, which are
strong for monocots. The selection efficiency (about 17%) was highest in the stem
and leaf explants after transformation with pGreen where nptII was under 35S.
About half of the selected plants were confirmed via PCR and Southern blot
analysis to be transgenic and, depending on the combination, 0 to 100% showed GUS
expression. GUS expression was strongest in multi-copy transgenic plants where
gus was under Act1. The same potato lines carrying multi-copy bar under Ubi1 were
also highly resistant to the herbicide Basta. The suggestion of using
Agrobacterium-mediated transformation of diploid lines of potato as a model crop
is discussed herein.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17160584 [PubMed - indexed for MEDLINE]

926: Transgenic Res. 2007 Oct;16(5):645-56. Epub 2006 Dec 8.

Fusion proteins comprising the catalytic domain of mutansucrase and a
starch-binding domain can alter the morphology of amylose-free potato starch
granules during biosynthesis.

Nazarian Firouzabadi F, Kok-Jacon GA, Vincken JP, Ji Q, Suurs LC, Visser RG.

Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding,
Wageningen University, 386, 6700 AJ Wageningen, The Netherlands.

It has been shown previously that mutan can be co-synthesized with starch when a
truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The
mutan seemed to adhere to the isolated starch granules, but it was not
incorporated in the starch granules. In this study, GtfICAT was fused to the N-
or C-terminus of a starch-binding domain (SBD). These constructs were introduced
into two genetically different potato backgrounds (cv. Kardal and amf), in order
to bring GtfICAT in more intimate contact with growing starch granules, and to
facilitate the incorporation of mutan polymers in starch. Fusion proteins of the
appropriate size were evidenced in starch granules, particularly in the amf
background. The starches from the various GtfICAT/SBD transformants seemed to
contain less mutan than those from transformants with GtfICAT alone, suggesting
that the appended SBD might inhibit the activity of GtfICAT in the engineered
fusion proteins. Scanning electron microscopy showed that expression of
SBD-GtfICAT resulted in alterations of granule morphology in both genetic
backgrounds. Surprisingly, the amf starches containing SBD-GtfICAT had a
spongeous appearance, i.e., the granule surface contained many small holes and
grooves, suggesting that this fusion protein can interfere with the lateral
interactions of amylopectin sidechains. No differences in physico-chemical
properties of the transgenic starches were observed. Our results show that
expression of granule-bound and "soluble" GtfICAT can affect starch biosynthesis
differently.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17160452 [PubMed - indexed for MEDLINE]

927: Nat Biotechnol. 2006 Dec;24(12):1472-3.

Comment in:
Nat Biotechnol. 2007 May;25(5):505-6; author reply 506.

Comment on:
Nat Biotechnol. 2006 Apr;24(4):435-6.

Why the omega-3 piggy should not go to market.

Fiester A.

Publication Types:
Comment
Letter

PMID: 17160035 [PubMed - indexed for MEDLINE]

928: Genes Genet Syst. 2006 Oct;81(5):349-54.

Overexpression of wheat alternative oxidase gene Waox1a alters respiration
capacity and response to reactive oxygen species under low temperature in
transgenic Arabidopsis.

Sugie A, Naydenov N, Mizuno N, Nakamura C, Takumi S.

Laboratory of Plant Genetics, Faculty of Agriculture, and Graduate School of
Science and Technology, Kobe University, Kobe, Japan.

Under low temperature conditions, the cytochrome pathway of respiration is
repressed and reactive oxygen species (ROS) are produced in plants. Mitochondrial
alternative oxidase (AOX) is the terminal oxidase responsible for the
cyanide-insensitive and salicylhydroxamic acid-sensitive respiration. To study
functions of wheat AOX genes under low temperature, we produced transgenic
Arabidopsis by introducing Waox1a expressed under control of the cauliflower
mosaic virus (CaMV) 35S promoter in Arabidopsis thaliana. The enhancement of
endogenous AOX1a expression via low temperature stress was delayed in the
transgenic Arabidopsis. Recovery of the total respiration activity under low
temperature occurred more rapidly in the transgenic plants than in the wild-type
plants due to a constitutively increased alternative pathway capacity. Levels of
ROS decreased in the transgenic plants under low temperature stress. These
results support the hypothesis that AOX alleviates oxidative stress when the
cytochrome pathway of respiration is inhibited under abiotic stress conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17159296 [PubMed - indexed for MEDLINE]

929: Appl Environ Microbiol. 2007 Feb;73(3):939-46. Epub 2006 Dec 8.

Expression of a novel small antimicrobial protein from the seeds of motherwort
(Leonurus japonicus) confers disease resistance in tobacco.

Yang X, Xiao Y, Wang X, Pei Y.

Biotechnology Research Center, Southwest Agricultural University, 400716
Chongqing, China. yangxy94@swu.edu.cn

Medicinal plants are valuable resources of natural antimicrobial materials. A
novel small protein with antimicrobial activities, designated LJAMP1, was
purified from the seeds of a medicinal herb, motherwort (Leonurus japonicus
Houtt). LJAMP1 is a heat-stable protein with a molecular mass of 7.8 kDa and a
determined isoelectric point of 8.2. In vitro assays showed that LJAMP1 inhibits
the growth of an array of fungi and bacteria. The hyphal growth inhibition by
LJAMP1 was more evident against hyphomycete fungi, such as Alternaria alternata,
Cercospora personata, and Aspergillus niger. The N-terminal amino acid sequence
of LJAMP1 was determined, and its coding gene was consequently cloned by the
rapid amplification of cDNA ends. The gene LJAMP1 has no intron and encodes a
polypeptide of 95 amino acids, in which the first 27 residues was deduced as a
signal peptide. The mature LJAMP1 shows relatively low identity to plant
napin-like storage proteins. Northern blot assays revealed that LJAMP1 is
expressed preferentially in seeds. Bioassays in transgenic tobacco demonstrated
that that overexpression of LJAMP1 significantly enhanced the resistance of
tobacco against not only the fungal pathogen A. alternata but also the bacterial
pathogen Ralstonia solanacearum, while no visible alteration in plant growth and
development was observed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17158620 [PubMed - indexed for MEDLINE]

930: J Biotechnol. 2007 Feb 1;128(2):383-92. Epub 2006 Nov 3.

Isolation and some effects of functional, low-phenylalanine kappa-casein
expressed in the milk of transgenic rabbits.

Baranyi M, Hiripi L, Szabö¨ L, Catunda AP, Harsö¡nyi I, Komö¡romy P, Bosze Z.

Agricultural Biotechnology Center, H-2100 Gö¶dö¶llo, Szent-Gyö¶rgyi A. u. 4,
Hungary.

Patients suffering certain metabolic diseases (e.g. phenylketonuria) need a
low-phenylalanine diet throughout their lives. Transgenic rabbits were created to
express low-phenylalanine kappa-casein in their milk. The aim was to demonstrate
for the first time the feasibility of producing a modified milk protein in
addition to normal milk proteins. A gene construct containing the coding region
of the rabbit kappa-casein gene was modified by site-specific oligonucleotide
directed mutagenesis. Four of the five phenylalanine amino acids present in the
mature protein were mutated and the gene construct was used to create two
transgenic rabbit lines. The transgenic rabbits produced the recombinant
kappa-casein at a high level in their milk causing a reduction in the average
size of the casein micelles. The low-phenylalanine kappa-casein was digestible
with chymosin and it was separated from its native counterpart and from the other
milk proteins by a one-step HPLC method on a reversed-phase column. In the
future, low-phenylalanine casein produced in transgenic animals could be used as
dietary replacements to meet the special requirements of certain consumer groups.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17157946 [PubMed - indexed for MEDLINE]

931: Behav Brain Res. 2007 Feb 12;177(1):22-9. Epub 2006 Dec 8.

Spatial learning in Long-Evans Hooded rats and C57BL/6J mice: different
strategies for different performance.

Cressant A, Besson M, Suarez S, Cormier A, Granon S.

Laboratoire de Physiologie de la Perception et de l'Action, UMR CNRS 7124,
Collö¨ge de France, Paris, France.

Spatial learning abilities of rodents have been extensively used to explore the
management of a wide range of cognitive and emotional processes such as learning,
memory, attention and anxiety. Knowledge about the organization and processing of
spatial learning has mainly been obtained in rats. Due to increasing generation
of genetically modified mice, cognitive abilities of mice are now extensively
tested. The present paper aimed at comparing spatial representation, learning and
strategies in C57BL/6J mice and Long-Evans Hooded rats when subjected to the same
spatial learning paradigm, i.e. learning a food location in a crossmaze. We also
analyzed the influence of environmental richness on learning modalities in both
species. Our results showed that rats and mice could exhibit similar spatial
learning abilities in some circumstances. However, Long-Evans rats and C57BL/6J
mice may set up different strategies depending on the availability of visual
information within the environment. Rats' learning strategies mainly relied on
distant visual cues and seemed more efficient than those used by mice as they
needed less time than mice to solve the task. We emphasize that the strategies of
mice are less robust and flexible than the ones set up by rats. Finally, the
richness of the environment was shown to affect speed and quality of spatial
learning in both species.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17157932 [PubMed - indexed for MEDLINE]

932: Metab Eng. 2007 Jan;9(1):95-111. Epub 2006 Oct 4.

Novel transgenic rice overexpressing anthocyanidin synthase accumulates a mixture
of flavonoids leading to an increased antioxidant potential.

Reddy AM, Reddy VS, Scheffler BE, Wienand U, Reddy AR.

Plant Molecular Genetics and Functional Genomics Laboratory, Department of Plant
Sciences, School of Life Sciences, University of Hyderabad, AP 500 046, India.
a_mmreddy@yahoo.co.in

In addition to their plant-associated functions, flavonoids act as antioxidants
against harmful free radicals in animals. Genetic engineering of food crops for a
mix of antioxidant flavonoids is highly beneficial in promoting human health.
Anthocyanidin synthase (ANS) is one of the four dioxygenases (DOX) of the
flavonoid biosynthetic pathway that catalyzes the formation of anthocyanidins
from leucoanthocyanidins. To investigate whether ANS mediates different DOX
reactions of the pathway and produces a mix of flavonoids, the rice ANS cDNA was
cloned and overexpressed in a rice mutant Nootripathu (NP). This mutant
accumulates proanthocyanidins exclusively in pericarp and absolutely no
anthocyanins in any tissue. In silico sequence analysis revealed that ANS
contains a double-stranded beta helix and shows high sequence similarity with
other DOXs of the pathway including flavonol synthase, flavonone
3beta-hydroxylase and flavone synthase I. Bacterially expressed ANS protein
converted dihydroquercetin to quercetin and Pro(35S):ANS complemented the maize
a2 mutant in producing anthocyanins in aleurone, suggesting that ANS functions as
a DOX with different flavonoid substrates. Similarly, transgenic NP plants
overexpressing Pro(MAS):ANS channeled the proanthocaynidin precursors to the
production of anthocyanins in pericarp. Transgenics showed approximately ten and
four-fold increase in the ANS transcripts and enzyme activity, respectively. As a
result, these plants showed an increased accumulation of a mixture of flavonoids
and anthocyanins, with a concomitant decrease in proanthocyanidins, suggesting
that ANS may act directly on different flavonoid substrates of DOX reactions.
Thus, overexpression of ANS in a rice mutant resulted in novel transgenic rice
with a mixture of flavonoids and an enhanced antioxidant potential.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17157544 [PubMed - indexed for MEDLINE]

933: Biotechnol Adv. 2007 Jan-Feb;25(1):75-84. Epub 2006 Oct 24.

Herbicide resistance of transgenic rice plants expressing human CYP1A1.

Kawahigashi H, Hirose S, Ohkawa H, Ohkawa Y.

Plant Biotechnology Department, National Institute of Agrobiological Sciences,
2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan. hiwak@affrc.go.jp

Cytochrome P450 monooxygenases (P450s) metabolize herbicides to produce mainly
non-phytotoxic metabolites. Although rice plants endogenously express multiple
P450 enzymes, transgenic plants expressing other P450 isoforms might show
improved herbicide resistance or reduce herbicide residues. Mammalian P450s
metabolizing xenobiotics are reported to show a broad and overlapping substrate
specificity towards lipophilic foreign chemicals, including herbicides. These
P450s are ideal for enhancing xenobiotic metabolism in plants. A human P450,
CYP1A1, metabolizes various herbicides with different structures and modes of
herbicide action. We introduced human CYP1A1 into rice plants, and the transgenic
rice plants showed broad cross-resistance towards various herbicides and
metabolized them. The introduced CYP1A1 enhanced the metabolism of chlorotoluron
and norflurazon. The herbicides were metabolized more rapidly in the transgenic
rice plants than in non-transgenic controls. Transgenic rice plants expressing
P450 might be useful for reducing concentrations of various chemicals in the
environment.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17156966 [PubMed - indexed for MEDLINE]

934: Anat Histol Embryol. 2006 Dec;35(6):351-6.

Ultrastructural morphometry of mammary gland in transgenic and non-transgenic
rabbits.

Dragin S, Pivko J, Massanyi P, Lukac N, Makarevich AV, Paleyanda RK, Chrenek P.

Slovak Agricultural Research Authority, Nitra, Slovak Republic.

The mammary gland of transgenic animals has been used for the production of
recombinant proteins of therapeutic and nutraceutical use. The objective of this
study was to compare the ultrastructure of transgenic and non-transgenic rabbit
mammary gland tissue. New Zealand White transgenic rabbits were obtained by
breeding non-transgenic rabbits with transgenic founder rabbits containing a whey
acidic protein-human factor VIII (WAP-hFVIII) transgene integrated into their
genome. Samples of mammary gland tissue from lactating rabbit females were
isolated by surgical procedures. These samples were examined by optical and
electron microscopy and photographs were taken. Measurements of ultrastructural
organelles were made from digital images of the mammary cells. No differences
were found in the cellular structure of mammary tissue, but significant
differences t((0.001)) in the relative volume of mitochondria and vacuoles
between transgenic and non-transgenic mammary gland epithelium were observed.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17156086 [PubMed - indexed for MEDLINE]

935: Biosci Biotechnol Biochem. 2006 Dec;70(12):2965-73. Epub 2006 Dec 7.

Rapid quantification methods for genetically modified maize contents using
genomic DNAs pretreated by sonication and restriction endonuclease digestion for
a capillary-type real-time PCR system with a plasmid reference standard.

Toyota A, Akiyama H, Sugimura M, Watanabe T, Sakata K, Shiramasa Y, Kitta K, Hino
A, Esaka M, Maitani T.

Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima.

For rough quantitative analysis of genetically modified maize contents, rapid
methods for measurement of the copy numbers of the cauliflower mosaic virus 35S
promoter region (P35S) and MON810 construct-specific gene (MON810) using a
combination of a capillary-type real-time PCR system with a plasmid DNA were
established. To reduce the characteristic differences between the plasmid DNA and
genomic DNA, we showed that pretreatment of the extracted genomic DNA by a
combination of sonication and restriction endonuclease digestion before
measurement is effective. The accuracy and reproducibility of this method for
MON810 content (%) at a level of 5.0% MON810 mixed samples were within a range
from 4.26 to 5.11% in the P35S copy number quantification. These methods should
prove to be a useful tool to roughly quantify GM maize content.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17151472 [PubMed - indexed for MEDLINE]

936: FEBS Lett. 2006 Dec 22;580(30):6891-7. Epub 2006 Nov 29.

Transport of antimony salts by Arabidopsis thaliana protoplasts over-expressing
the human multidrug resistance-associated protein 1 (MRP1/ABCC1).

Gayet L, Picault N, Cazalö© AC, Beyly A, Lucas P, Jacquet H, Suso HP, Vavasseur A,
Peltier G, Forestier C.

CEA Cadarache, DSV-DEVM--LEMS, UMR 6191 CNRS-CEA-Universitö© Aix-Marseille II,
13108 St Paul lez Durance, France.

ABC transporters from the multidrug resistance-associated protein (MRP) subfamily
are glutathione S-conjugate pumps exhibiting a broad substrate specificity
illustrated by numerous xenobiotics, such as anticancer drugs, herbicides,
pesticides and heavy metals. The engineering of MRP transporters into plants
might be interesting either to reduce the quantity of xenobiotics taken up by the
plant in the context of "safe-food" strategies or, conversely, in the development
of phytoremediation strategies in which xenobiotics are sequestered in the
vacuolar compartment. In this report, we obtained Arabidopsis transgenic plants
overexpressing human MRP1. In these plants, expression of MRP1 did not increase
plant resistance to antimony salts (Sb(III)), a classical glutathione-conjugate
substrate of MRP1. However, the transporter was fully translated in roots and
shoots, and targeted to the plasma membrane. In order to investigate the
functionality of MRP1 in Arabidopsis, mesophyll cell protoplasts (MCPs) were
isolated from transgenic plants and transport activities were measured by using
calcein or Sb(III) as substrates. Expression of MRP1 at the plasma membrane was
correlated with an increase in the MCPs resistance to Sb(III) and a limitation of
the metalloid content in the protoplasts due to an improvement in Sb(III) efflux.
Moreover, Sb(III) transport was sensitive to classical inhibitors of the human
MRP1, such as MK571 or glibenclamide. These results demonstrate that a human ABC
transporter can be functionally introduced in Arabidopsis, which might be useful,
with the help of stronger promoters, to reduce the accumulation of xenobiotics in
plants, such as heavy metals from multi-contaminated soils.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17150215 [PubMed - indexed for MEDLINE]

937: Plant Mol Biol. 2007 Mar;63(4):571-88.

Role of a novel pathogen-induced pepper C3-H-C4 type RING-finger protein gene,
CaRFPI, in disease susceptibility and osmotic stress tolerance.

Hong JK, Choi HW, Hwang IS, Hwang BK.

Laboratory of Molecular Plant Pathology, College of Life Sciences and
Biotechnology, Korea University, Anam-dong, Sungbuk-ku, Seoul 136-713, Republic
of Korea.

Limited information is available about the roles of RING-finger proteins in plant
defense. A pepper CaRFP1 encoding the C3-H-C4 type RING-finger protein that
physically interacted with the basic PR-1 protein CABPR1 was isolated from pepper
leaves infected by Xanthomonas campestris pv. vesicatoria. The CaRFP1 protein has
VWFA domain, and N-terminal serine-rich and C-terminal cysteine-rich regions. The
CaRFP1 transcripts accumulated earlier than did those of the basic PR-1 gene
CABPR1 during the incompatible interaction of pepper leaves with X. campestris
pv. vesicatoria, as well as in the systemic, uninoculated pepper leaf tissues.
The CaRFP1 gene also was induced in pepper leaf tissues infected by
Colletotrichum coccodes. The CaRFP1 gene was strongly induced much earlier by
salicylic acid, ethylene and methyl jasmonate treatments, as well as
environmental stresses including methyl viologen, mannitol and NaCl treatments.
Overexpression of the CaRFP1 gene in the transgenic Arabidopsis plants conferred
disease susceptibility to Pseudomonas syringae pv. tomato infection, accompanied
by reduced PR-2 and PR-5 gene expression, suggesting that the CaRFP1 acts as an
E3 ligase for polyubiquitination of target PR proteins. Exogenous salicylic acid
treatment also abolished PR-2 and PR-5 gene expression in the transgenic plants.
Differential osmotic stress tolerance was induced by high salt and drought in the
CaRFPI-overexpressing plants during germination and seedling development, which
was closely correlated with abscisic acid sensitivity of Arabidopsis plants.
These results suggest that the CaRFP1 gene functions as an early defense
regulator controlling bacterial disease susceptibility and osmotic stress
tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17149652 [PubMed - indexed for MEDLINE]

938: Plant Cell Rep. 2007 May;26(5):611-5. Epub 2006 Dec 6.

Delay of leaf senescence in Medicago sativa transformed with the ipt gene
controlled by the senescence-specific promoter SAG12.

Calderini O, Bovone T, Scotti C, Pupilli F, Piano E, Arcioni S.

CNR Istituto di Genetica Vegetale, via Madonna Alta 130, Perugia, Italy.

We report the successfull delay of leaf senescence in Medicago sativa. A highly
regenerable clone of alfalfa was transformed with the construct SAG12-IPT, an
approach that has already proved efficient in other crops. Several independent
transformants were obtained as determined by Southern analysis and all the
transformants expressed the transgene as measured by RT-PCR. In vitro and in vivo
analyses showed that SAG12-IPT plants exhibited a stay-green phenotype that has
the potential to greatly improve the quantity and quality of alfalfa forage.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17149639 [PubMed - indexed for MEDLINE]

939: Biol Lett. 2006 Jun 22;2(2):198-202.

Fees or refuges: which is better for the sustainable management of insect
resistance to transgenic Bt corn?

Vacher C, Bourguet D, Desquilbet M, Lemariö© S, Ambec S, Hochberg ME.

Universitö© Montpellier II, Institut des Sciences de l'Evolution de Montpellier
(UMR 5554), 34095 Montpellier Cedex 05, France. vacher@antibes.inra.fr

The evolution of resistance in insect pests will imperil the efficiency of
transgenic insect-resistant crops. The currently advised strategy to delay
resistance evolution is to plant non-toxic crops (refuges) in close proximity to
plants engineered to express the toxic protein of the bacterium Bacillus
thuringiensis (Bt). We seek answers to the question of how to induce growers to
plant non-toxic crops. A first strategy, applied in the United States, is to
require Bt growers to plant non-Bt refuges and control their compliance with
requirements. We suggest that an alternative strategy is to make Bt seed more
expensive by instituting a user fee, and we compare both strategies by
integrating economic processes into a spatially explicit, population genetics
model. Our results indicate that although both strategies may allow the
sustainable management of the common pool of Bt-susceptibility alleles in pest
populations, for the European corn borer (Ostrinia nubilalis) one of the most
serious pests in the US corn belt, the fee strategy is less efficient than refuge
requirements.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17148361 [PubMed - indexed for MEDLINE]

940: Transgenic Res. 2007 Apr;16(2):177-91. Epub 2006 Dec 5.

The GUS reporter-aided analysis of the promoter activities of a rice
metallothionein gene reveals different regulatory regions responsible for
tissue-specific and inducible expression in transgenic Arabidopsis.

Lö¼ S, Gu H, Yuan X, Wang X, Wu AM, Qu L, Liu JY.

Laboratory of Molecular Biology and Protein Science Laboratory of the Ministry of
Education, Department of Biological Sciences and Biotechnology, Tsinghua
University, Beijing, 100084, China.

To gain a better understanding of the regulatory mechanism of plant
metallothionein (MT) genes, a chimeric expression unit consisting of the
beta-glucuronidase (gusA) reporter gene under the control of a 1,324 bp fragment
of the rice MT (ricMT) promoter was introduced into Arabidopsis via Agrobacterium
tumefaciens. The strongest histochemical staining for GUS activity was observed
in the cotyledons and hypocotyls of the transgenic seedlings and in the stigma,
filaments and anthers of young and mature flowers, and especially in the wounded
tissues of transgenic plants. In contrast, a relatively low level of reporter
gene expression was seen in the young roots of transgenic seedlings and no GUS
activity was detected in the stems, seeds and leaves, but GUS activity was
observed in cotyledons and the first two true leaves. Promoter analysis of 5'
deletions further identified several important regions responsible for
organ-specific expression including roots, flowers and wound induction, light and
ABA, Cu and Zn responses. These findings demonstrate that a 1,324 bp fragment of
the rice MT promoter performs a complicated transcriptional regulation with
clearly functional regions in a model plant, and provide an important insight
into the transcriptional regulation mechanisms that operate the temporal- and
spatial-specific expression and stress responses of the rice MT gene. These
results suggest that the ricMT promoter and its functional regions are
potentially useful in genetic engineering of plants to express the desired genes
whose products are preferentially needed in roots, flowers and wound induction.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17146614 [PubMed - indexed for MEDLINE]

941: An Acad Bras Cienc. 2006 Dec;78(4):667-86.

GMOs: building the future on the basis of past experience.

Reis LF, Van Sluys MA, Garratt RC, Pereira HM, Teixeira MM.

Ludwig Institute for Cancer Research, Sö¸o Paulo, SP, Brazil. lreis@ludwig.org.br

Biosafety of genetically modified organisms (GMOs) and their derivatives is still
a major topic in the agenda of government and societies worldwide. The aim of
this review is to bring into light that data that supported the decision taken
back in 1998 as an exercise to stimulate criticism from the scientific community
for upcoming discussions and to avoid emotional and senseless arguments that
could jeopardize future development in the field. It must be emphasized that
Roundup Ready soybean is just one example of how biotechnology can bring in
significant advances for society, not only through increased productivity, but
also with beneficial environmental impact, thereby allowing more rational use of
agricultural pesticides for improvement of the soil conditions. The adoption of
agricultural practices with higher yield will also allow better distribution of
income among small farmers. New species of genetically modified plants will soon
be available and society should be capable of making decisions in an objective
and well-informed manner, through collegiate bodies that are qualified in all
aspects of biosafety and environmental impact.

Publication Types:
Review

PMID: 17143405 [PubMed - indexed for MEDLINE]

942: Plant Physiol. 2007 Feb;143(2):570-8. Epub 2006 Dec 1.

Transposition-based plant transformation.

Yan H, Rommens CM.

Simplot Plant Sciences, J.R. Simplot Company, Boise, Idaho 83706, USA.

Agrobacterium T-DNAs were used to deliver transposable Dissociation (Ds) elements
into the nuclei of potato (Solanum tuberosum) cells. A double-selection system
was applied to enrich for plants that only contained a transposed Ds element.
This system consisted of a positive selection for the neomycin phosphotransferase
(nptII) gene positioned within Ds followed by a negative selection against stable
integration of the cytosine deaminase (codA) gene-containing T-DNA. Sixteen of 29
transgenic plants were found to contain a transposed element while lacking any
superfluous T-DNA sequences. The occurrence of this genotype indicates that Ds
elements can transpose from relatively short extrachromosomal DNA molecules into
the plant genome. The frequency of single-copy Ds transformation was determined
at 0.3%, which is only about 2.5-fold lower than the potato transformation
frequency for backbone-free and single-copy T-DNAs. Because of the generally high
expression levels of genes positioned within transposed elements, the new
transformation method may find broad applicability to crops that are accessible
to Agrobacterium T-DNA transfer.

PMID: 17142486 [PubMed - indexed for MEDLINE]

943: Plant Physiol. 2007 Feb;143(2):650-60. Epub 2006 Dec 1.

Cell death suppressor Arabidopsis bax inhibitor-1 is associated with calmodulin
binding and ion homeostasis.

Ihara-Ohori Y, Nagano M, Muto S, Uchimiya H, Kawai-Yamada M.

Institute of Molecular and Cellular Biosciences, The University of Tokyo,
Bunkyo-ku, Tokyo 113-0032, Japan.

Cell death suppressor Bax inhibitor-1 (BI-1), an endoplasmic reticulum membrane
protein, exists in a wide range of organisms. The split-ubiquitin system, overlay
assay, and bimolecular fluorescence complementation analysis demonstrated that
Arabidopsis (Arabidopsis thaliana) BI-1 (AtBI-1) interacted with calmodulin in
yeast (Saccharomyces cerevisiae) and in plant cells. Furthermore, AtBI-1 failed
to rescue yeast mutants lacking Ca2+ ATPase (Pmr1 or Spf1) from Bax-induced cell
death. Pmr1 and Spf1, p-type ATPases localized at the inner membrane, are
believed to be involved in transmembrane movement of calcium ions in yeast. Thus,
the presence of intact Ca2+ ATPases was essential for AtBI-1-mediated cell death
suppression in yeast. To investigate the effect of AtBI-1 on calcium homeostasis,
we evaluated sensitivity against cyclopiazonic acid (CPA), an inhibitor of
sarcoplasmic/endoplasmic reticulum Ca2+ ATPase in AtBI-1-overexpressing or
knock-down transgenic Arabidopsis plants. These plants demonstrated altered CPA
or ion stress sensitivity. Furthermore, AtBI-1-overexpressing cells demonstrated
an attenuated rise in cytosolic calcium following CPA or H2O2 treatment,
suggesting that AtBI-1 affects ion homeostasis in plant cell death regulation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17142482 [PubMed - indexed for MEDLINE]

944: Environ Pollut. 2007 Jun;147(3):540-5. Epub 2006 Dec 4.

Tolerance of transgenic canola plants (Brassica napus) amended with plant
growth-promoting bacteria to flooding stress at a metal-contaminated field site.

Farwell AJ, Vesely S, Nero V, Rodriguez H, McCormack K, Shah S, Dixon DG, Glick
BR.

Department of Biology, University of Waterloo, Waterloo, Ontario, Canada.
afarwell@sciborg.uwaterloo.ca

The growth of transgenic canola (Brassica napus) expressing a gene for the enzyme
1-aminocyclopropane-1-carboxylate (ACC) deaminase was compared to non-transformed
canola exposed to flooding and elevated soil Ni concentration, in situ. In
addition, the ability of the plant growth-promoting bacterium Pseudomonas putida
UW4, which also expresses ACC deaminase, to facilitate the growth of
non-transformed and transgenic canola under the above mentioned conditions was
examined. Transgenic canola and/or canola treated with P. putida UW4 had greater
shoot biomass compared to non-transformed canola under low flood-stress
conditions. Under high flood-stress conditions, shoot biomass was reduced and Ni
accumulation was increased in all instances relative to low flood-stress
conditions. This is the first field study to document the increase in plant
tolerance utilizing transgenic plants and plant growth-promoting bacteria exposed
to multiple stressors.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17141927 [PubMed - indexed for MEDLINE]

945: Mycologia. 2006 Jul-Aug;98(4):593-7.

An optimized method for mycelial compatibility testing in Sclerotinia
sclerotiorum.

Schafer MR, Kohn LM.

Biology Department, University of Toronto at Mississauga, 3359 Mississauga Road
North, Mississauga, ON, L5L 1C6, Canada.

Classification of isolates into mycelial compatibility groups (MCGs) is used
routinely in many laboratories as a quick marker for genotyping Sclerotinia
sclerotiorum within populations. Scoring each new sample requires optimization of
standardized conditions to support adequate growth of all paired isolates.
Appropriate conditions for growth are especially important because diverse
compatibility reactions are difficult to categorize and score (e.g., in samples
from populations with high genetic diversity, such as those that receive
immigration from genetically diverse sources or those that deviate from strict
clonality). The current standard medium for MCG testing can be inhibitory to
isolates from some samples, confounding scoring of compatibility. We identified
two foci for optimization: (i) choice of medium, in this experiment, Patterson's
medium amended with red food coloring (termed modified Patterson's medium, MPM,
the current standard medium) versus potato dextrose agar (PDA) and (ii) amount of
McCormick's red food coloring amended to the growth medium. The red food coloring
often yields a red reaction line in incompatible interactions; alternative
incompatible reactions are a line of thick or thin hyphae. Based on results to
date, self-self pairings of S. sclerotiorum are compatible and are a reliable
standard for scoring compatible self-nonself mycelial interactions. PDA amended
with 75 microl/L of McCormick's red food coloring was identified as optimal for
isolates inhibited by MPM from a highly diverse, recombining population sample.
This precisely amended PDA was also suitable for isolates from highly clonal
populations that were not inhibited by MPM or by higher concentrations of red
food coloring. Under the optimized, standardized conditions all paired isolates
grew together and produced interactions that could be scored in repeatedly
identifiable categories, compatible or incompatible. Workers are advised to
optimize conditions before screening a new population sample.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17139852 [PubMed - indexed for MEDLINE]

946: J Plant Res. 2007 Mar;120(2):237-45. Epub 2006 Dec 1.

Development of genotype-independent regeneration system for transformation of
rice (Oryza sativa ssp. indica).

Yookongkaew N, Srivatanakul M, Narangajavana J.

Department of Biotechnology, Faculty of Science, Mahidol University, Rama VI
Road, Bangkok, 10400, Thailand.

Rice (Oryza sativa ssp. indica) is an important economic crop in many countries.
Although a variety of conventional methods have been developed to improve this
plant, manipulation by genetic engineering is still complicated. We have
established a system of multiple shoot regeneration from rice shoot apical
meristem. By use of MS medium containing 4 mg L(-1) thidiazuron (TDZ) multiple
shoots were successfully developed directly from the meristem without an
intervening callus stage. All rice cultivars tested responded well on the medium
and regenerated to plantlets that were readily transferred to soil within 5-8
weeks. The tissue culture system was suitable for Agrobacterium-mediated
transformation and different factors affecting transformation efficiency were
investigated. Agrobacterium strain EHA105 containing the plasmid pCAMBIA1301 was
used. The lowest concentration of hygromycin B in combined with either 250 mg
L(-1) carbenicillin or 250 mg L(-1) cefotaxime to kill the rice shoot apical
meristem was 50 mg L(-1) and carbenicillin was more effective than cefotaxime.
Two-hundred micromolar acetosyringone had no effect on the efficiency of
transient expression. Sonication of rice shoot apical meristem for 10 s during
bacterial immersion increased transient GUS expression in three-day co-cultivated
seedlings. The gus gene was found to be integrated into the genome of the T(0)
transformant plantlets.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17139419 [PubMed - indexed for MEDLINE]

947: J Exp Bot. 2007;58(3):483-95. Epub 2006 Nov 30.

The role of the sucrose transporter, OsSUT1, in germination and early seedling
growth and development of rice plants.

Scofield GN, Aoki N, Hirose T, Takano M, Jenkins CL, Furbank RT.

CSIRO Plant Industry, Canberra, ACT 2601, Australia.

Using expression analysis, the role of the sucrose transporter OsSUT1 during
germination and early growth of rice seedlings has been examined in detail, over
a time-course ranging from 1 d to 7 d post-imbibition. Unlike the wheat
orthologue, TaSUT1, which is thought to be directly involved in sugar transfer
across the scutellar epithelium, OsSUT1 is not expressed in the scutellar
epithelial cell layer of germinating rice and is, therefore, not involved in
transport of sugars across the symplastic discontinuity between the endosperm and
the embryo. OsSUT1 expression was also absent from the aleurone cells, indicating
it is not involved in the transport of sucrose in this cell layer during
germination. However, by 3 d post-imbibition, OsSUT1 was present in the companion
cells and sieve elements of the scutellar vascular bundle, where it may play a
role in phloem loading of sucrose for transport to the developing shoot and
roots. This sucrose is most likely sourced from hexoses imported from the
endosperm. In addition, sucrose may be remobilized from starch granules which are
present at a high density in the scutellar ground tissues surrounding the
vasculature and at the base of the shoot. OsSUT1 was also present in the
coleoptile and the first and second leaf blades, where it was localized to the
phloem along the entire length of these tissues, and was also present within the
phloem of the primary roots. OsSUT1 may be involved in retrieval of sugars from
the apoplasm in these tissues.

PMID: 17138625 [PubMed - indexed for MEDLINE]

948: Plant Mol Biol. 2007 Mar;63(4):533-43.

The influence of matrix attachment regions on transgene expression in Arabidopsis
thaliana wild type and gene silencing mutants.

De Bolle MF, Butaye KM, Goderis IJ, Wouters PF, Jacobs A, Delaurö© SL, Depicker A,
Cammue BP.

Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven,
Kasteelpark 20, B-3001 Leuven, Belgium. miguel.debolle@biw.kuleuven.be

Many studies in both animal and plant systems have shown that matrix attachment
regions (MARs) can increase the expression of flanking transgenes. However, our
previous studies revealed no effect of the chicken lysozyme MARs (chiMARs) on
transgene expression in the first generation transgenic Arabidopsis thaliana
plants transformed with a beta-glucuronidase gene (uidA) unless gene silencing
mutants were used as genetic background for transformation. In the present study,
we investigated why chiMARs do not influence transgene expression in transgenic
wild-type Arabidopsis plants. We first studied the effect of chiMARs on transgene
expression in the progeny of primary transformants harboring chiMAR-flanked
T-DNAs. Our data indicate that chiMARs do not affect transgene expression in
consecutive generations of wild-type A. thaliana plants. Next, we examined
whether these observed results in A. thaliana transformants are influenced by the
applied transformation method. The results from in vitro transformed A. thaliana
plants are in accordance with those from in planta transformed A. thaliana plants
and again reveal no influence of chiMARs on transgene expression in A. thaliana
wild-type transformants. The effect of chi-MARs on transgene expression is also
examined in in vitro transformed Nicotiana tabacum plants, but as for A.
thaliana, the transgene expression in tobacco transformants is not altered by the
presence of chi-MARs. Taken together, our results show that the applied method or
the plant species used for transformation does not influence whether and how
chiMARs have an effect on transgene expression. Finally, we studied the effect of
MARs (tabMARs) of plant origin (tobacco) on the transgene expression in A.
thaliana wild-type plants and suppressed gene silencing (sgs2) mutants. Our
results clearly show that similar to chiMARs, the tobacco-derived MARs do not
enhance transgene expression in a wild-type background but can be used to enhance
transgene expression in a mutant impaired in gene silencing.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17136580 [PubMed - indexed for MEDLINE]

949: Mar Biotechnol (NY). 2007 Jan-Feb;9(1):92-100. Epub 2006 Nov 30.

Expression of masu salmon delta5-desaturase-like gene elevated EPA and DHA
biosynthesis in zebrafish.

Alimuddin , Yoshizaki G, Kiron V, Satoh S, Takeuchi T.

Department of Marine Biosciences, Tokyo University of Marine Science and
Technology, Minato, Tokyo 108-8477, Japan.

Farmed fish could substitute for marine capture fish as a source of fatty acids
such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) beneficial for
human health; however, they require these compounds in their diets. In the
present study on a model fish species, we modified the EPA/DHA biosynthesis
pathway by overexpression of masu salmon Delta5-desaturase-like gene in zebrafish
to increase its ability to synthesize EPA and DHA. Expression of this gene in
transgenic fish fed a commercial diet and Artemia helped to improve their EPA
content by 1.21-fold and DHA by 1.24-fold. In similar fish that were fed only
Artemia the increments were 1.14-fold for EPA and 1.13-fold for DHA, compared
with nontransgenic fish. In contrast, eicosatetraenoic acid content decreased, as
it is a substrate of Delta5-desaturase, while the total lipid remained constant.
The results demonstrated that masu salmon Delta5-desaturase is functional in
zebrafish and can modify its fatty acid metabolic pathway. The technique could be
applied to farmed fish to generate a nutritionally richer product for human
consumption.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17136489 [PubMed - indexed for MEDLINE]

950: Ann N Y Acad Sci. 2006 Oct;1081:1-16.

Biodiversity and emerging diseases.

Maillard JC, Gonzalez JP.

Cirad-Emvt/PRISE Hanoi, Vietnam. maillard@fpt.vn

First we remind general considerations concerning biodiversity on earth and
particularly the loss of genetic biodiversity that seems irreversible whether its
origin is directly or indirectly linked to human activities. Urgent and
considerable efforts must be made from now on to cataloge, understand, preserve,
and enhance the value of biodiversity while ensuring food safety and human and
animal health. Ambitious integrated and multifield research programs must be
implemented in order to understand the causes and anticipate the consequences of
loss of biodiversity. Such losses are a serious threat to sustainable development
and to the quality of life of future generations. They have an influence on the
natural balance of global biodiversity in particularly in reducing the capability
of species to adapt rapidly by genetic mutations to survive in modified
ecosystems. Usually, the natural immune systems of mammals (both human and
animal), are highly polymorphic and able to adapt rapidly to new situations. We
more specifically discuss the fact that if the genetic diversity of the affected
populations is low the invading microorganisms, will suddenly expand and create
epidemic outbreaks with risks of pandemic. So biodiversity appears to function as
an important barrier (buffer), especially against disease-causing organisms,
which can function in different ways. Finally, we discuss the importance of
preserving biodiversity mainly in the wildlife ecosystems as an integrated and
sustainable approach among others in order to prevent and control the emergence
or reemergence of diseases in animals and humans (zoonosis). Although plants are
also part of this paradigm, they fall outside our field of study.

Publication Types:
Review

PMID: 17135490 [PubMed - indexed for MEDLINE]

951: J Virol. 2007 Feb;81(4):1563-73. Epub 2006 Nov 29.

Tomato chlorotic mottle virus is a target of RNA silencing but the presence of
specific short interfering RNAs does not guarantee resistance in transgenic
plants.

Ribeiro SG, Lohuis H, Goldbach R, Prins M.

Laboratory of Virology, Binnenhaven 11, 6709 PD Wageningen, The Netherlands.

Tomato chlorotic mottle virus (ToCMoV) is a begomovirus found widespread in
tomato fields in Brazil. ToCMoV isolate BA-Se1 (ToCMoV-[BA-Se1]) was shown to
trigger the plant RNA silencing surveillance in different host plants and,
coinciding with a decrease in viral DNA levels, small interfering RNAs (siRNAs)
specific to ToCMoV-[BA-Se1] accumulated in infected plants. Although not
homogeneously distributed, the siRNA population in both infected Nicotiana
benthamiana and tomato plants represented the entire DNA-A and DNA-B genomes. We
determined that in N. benthamiana, the primary targets corresponded to the 5' end
of AC1 and the embedded AC4, the intergenic region and 5' end of AV1 and
overlapping central part of AC5. Subsequently, transgenic N. benthamiana plants
were generated that were preprogrammed to express double-stranded RNA
corresponding to this most targeted portion of the virus genome by using an
intron-hairpin construct. These plants were shown to indeed produce
ToCMoV-specific siRNAs. When challenge inoculated, most transgenic lines showed
significant delays in symptom development, and two lines had immune plants.
Interestingly, the levels of transgene-produced siRNAs were similar in resistant
and susceptible siblings of the same line. This indicates that, in contrast to
RNA viruses, the mere presence of transgene siRNAs corresponding to DNA virus
sequences does not guarantee virus resistance and that other factors may play a
role in determining RNA-mediated resistance to DNA viruses.

PMID: 17135316 [PubMed - indexed for MEDLINE]

952: Gene. 2007 Feb 15;388(1-2):1-13. Epub 2006 Oct 24.

Deciphering the regulatory mechanisms of abiotic stress tolerance in plants by
genomic approaches.

Sreenivasulu N, Sopory SK, Kavi Kishor PB.

Leibniz-Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse
3, 06466, Gatersleben, Germany. srinivas@ipk-gatersleben.de

Environmental constraints that include abiotic stress factors such as salt,
drought, cold and extreme temperatures severely limit crop productivity.
Improvement of crop plants with traits that confer tolerance to these stresses
was practiced using traditional and modern breeding methods. Molecular breeding
and genetic engineering contributed substantially to our understanding of the
complexity of stress response. Mechanisms that operate signal perception,
transduction and downstream regulatory factors are now being examined and an
understanding of cellular pathways involved in abiotic stress responses provide
valuable information on such responses. This review presents genomic-assisted
methods which have helped to reveal complex regulatory networks controlling
abiotic stress tolerance mechanisms by high-throughput expression profiling and
gene inactivation techniques. Further, an account of stress-inducible regulatory
genes which have been transferred into crop plants to enhance stress tolerance is
discussed as possible modes of integrating information gained from functional
genomics into knowledge-based breeding programs. In addition, we envision an
integrative genomic and breeding approach to reveal developmental programs that
enhance yield stability and improve grain quality under unfavorable environmental
conditions of abiotic stresses.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17134853 [PubMed - indexed for MEDLINE]

953: Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18842-7. Epub 2006 Nov 28.

Silencing leaf sorbitol synthesis alters long-distance partitioning and apple
fruit quality.

Teo G, Suzuki Y, Uratsu SL, Lampinen B, Ormonde N, Hu WK, DeJong TM, Dandekar AM.

Department of Plant Sciences, University of California-Davis, 1 Shields Avenue,
Davis, CA 95616, USA.

Sorbitol and sucrose are major products of photosynthesis distributed in apple
trees (Malus domestica Borkh. cv. "Greensleeves") that affect quality in fruit.
Transgenic apple plants were silenced or up-regulated for sorbitol-6-phosphate
dehydrogenase by using the CaMV35S promoter to define the role of sorbitol
distribution in fruit development. Transgenic plants with suppressed
sorbitol-6-phosphate dehydrogenase compensated by accumulating sucrose and starch
in leaves, and morning and midday net carbon assimilation rates were
significantly lower. The sorbitol to sucrose ratio in leaves was reduced by
approximately 90% and in phloem exudates by approximately 75%. The fruit
accumulated more glucose and less fructose, starch, and malic acid, with no
overall differences in weight and firmness. Sorbitol dehydrogenase activity was
reduced in silenced fruit, but activities of neutral invertase, vacuolar
invertase, cell wall-bound invertase, fructose kinase, and hexokinase were
unaffected. Analyses of transcript levels and activity of enzymes involved in
carbohydrate metabolism throughout fruit development revealed significant
differences in pathways related to sorbitol transport and breakdown. Together,
these results suggest that sorbitol distribution plays a key role in fruit carbon
metabolism and affects quality attributes such as sugar-acid balance and starch
accumulation.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 17132742 [PubMed - indexed for MEDLINE]

954: Carcinogenesis. 2006 Dec;27(12):2555-64.

Corrected and republished from:
Carcinogenesis. 2006 Oct;27(10):1970-9.

Dietary effects of soy isoflavones daidzein and genistein on
7,12-dimethylbenz[a]anthracene-induced mammary mutagenesis and carcinogenesis in
ovariectomized Big Blue transgenic rats.

Manjanatha MG, Shelton S, Bishop ME, Lyn-Cook LE, Aidoo A.

Division of Genetic and Reproductive Toxicology, National Center for
Toxicological Research, U.S. FDA Jefferson Laboratories, Jefferson, AR 72079,
USA. mmanjanatha@nctr.fda.gov

The major constituents of isoflavones, daidzein (DZ) and genistein (GE) are known
to interact with the alpha and beta estrogen receptors (ERalpha/beta) in several
tissues including mammary. In this study, we used ovariectomy (OVX) to model
menopause and determined the effects of DZ, GE or 17beta-estradiol (E2) exposures
on chemically induced mutagenesis and carcinogenesis in the mammary glands of
female Big Blue (BB) transgenic rats. The rats were fed control diet containing
the isoflavones and E2 and treated with a single oral dose of
7,12-dimethylbenz[a]anthracene (DMBA) at PND 50. Animals were sacrificed at 16 or
20 weeks post-carcinogen treatment to assess mutant frequencies (MFs) and
histopathological parameters, respectively. The isoflavones or E2 supplementation
alone resulted in modest increases in the lacI MF that were not significantly
different from the MFs measured in rats fed the control diet alone. DMBA
exposure, however, induced significant increases in the lacI MFs in the mammary
of both OVX and ovary intact (INT) rats and Hprt MFs in spleen lymphocytes
(P any significant changes in DMBA-induced mutagenicity in the mammary. However,
feeding the isoflavone mixture (DZG) resulted in a significant reduction in the
DMBA-induced lacI MFs (P immunohistochemistry was increased in both OVX and INT rats exposed to DMBA as
compared with rats fed control diet (P hyperplasia was induced in most of the treatment groups including control.
Although DMBA treatment did not induce mammary tumors in the OVX rats, adenoma
and adenocarcinoma were detected in the mammary glands of INT rats.

Publication Types:
Corrected and Republished Article
Research Support, U.S. Gov't, P.H.S.

PMID: 17127718 [PubMed - indexed for MEDLINE]

955: Genomics Proteomics Bioinformatics. 2006 Aug;4(3):137-44.

Functional characterization of gibberellin-regulated genes in rice using
microarray system.

Jan A, Komatsu S.

National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan.

Gibberellin (GA) is collectively referred to a group of diterpenoid acids, some
of which act as plant hormones and are essential for normal plant growth and
development. DNA microarray technology has become the standard tool for the
parallel quantification of large numbers of messenger RNA transcripts. The power
of this approach has been demonstrated in dissecting plant physiology and
development, and in unraveling the underlying cellular signaling pathways. To
understand the molecular mechanism by which GA regulates the growth and
development of plants, with reference to the monocot model plant-rice, it is
essential to identify and analyze more genes and their products at the
transcription and translation levels that are regulated by GA. With the
availability of draft sequences of two major rice types, indica and japonica
rice, it has become possible to analyze global expression profiles of genes on a
genome scale. In this review, the progress made in finding new genes in rice leaf
sheath using microarray system and their characterization is discussed. It is
believed that the findings made in this regard have important implications for
understanding the mechanism by which GA regulates the growth and development of
rice.

Publication Types:
Review

PMID: 17127211 [PubMed - indexed for MEDLINE]

956: Biotechnol J. 2006 Dec;1(12):1433-4.

Consumer acceptance of ingenic foods.

Lusk JL, Rozan A.

Department of Agricultural Economics, Oklahoma State University, Stillwater, OK,
USA. jayson.lusk@okstate.edu

Recent advances in plant molecular biology offer a means of reaping the benefits
of biotechnology, while potentially assuaging consumer concerns by re-inserting
native DNA back into plants. Results are presented from nationwide surveys in the
U.S. and France, indicating that more consumers would accept ingenic plants than
transgenic plants, with twice as many U.S. than French consumers considering food
produced through biotechnology eatable.

PMID: 17124706 [PubMed - indexed for MEDLINE]

957: Science. 2006 Nov 24;314(5803):1298-301.

Comment in:
Science. 2006 Nov 24;314(5803):1252-3.

A NAC Gene regulating senescence improves grain protein, zinc, and iron content
in wheat.

Uauy C, Distelfeld A, Fahima T, Blechl A, Dubcovsky J.

Department of Plant Sciences, University of California, One Shields Avenue,
Davis, CA 95616, USA.

Enhancing the nutritional value of food crops is a means of improving human
nutrition and health. We report here the positional cloning of Gpc-B1, a wheat
quantitative trait locus associated with increased grain protein, zinc, and iron
content. The ancestral wild wheat allele encodes a NAC transcription factor
(NAM-B1) that accelerates senescence and increases nutrient remobilization from
leaves to developing grains, whereas modern wheat varieties carry a nonfunctional
NAM-B1 allele. Reduction in RNA levels of the multiple NAM homologs by RNA
interference delayed senescence by more than 3 weeks and reduced wheat grain
protein, zinc, and iron content by more than 30%.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17124321 [PubMed - indexed for MEDLINE]

958: J Exp Bot. 2006;57(15):4155-69. Epub 2006 Nov 22.

Cloning and characterization of Arabidopsis and Brassica juncea flavin-containing
amine oxidases.

Lim TS, Chitra TR, Han P, Pua EC, Yu H.

Department of Biological Sciences and Temasek Life Sciences Laboratory, National
University of Singapore, Singapore 117543.

Polyamines (PAs) are low molecular weight metabolites involved in various
physiological and developmental processes in eukaryotic and prokaryotic cells.
The cellular PA level is regulated in part by the action of amine oxidases (AOs)
including copper diamine oxidases (DAOs) and flavoprotein polyamine oxidases
(PAOs). In this study, the isolation and characterization of flavin amine
oxidases (FAOs) from Brassica juncea (BJFAO) and Arabidopsis (ATFAO1) are
reported that were clustered in the same group as polyamine oxidases from maize
(MPAO) and barley (BPAO1) and monoamine oxidases from mammalian species. ATFAO1
was temporally and spatially regulated in Arabidopsis and showed distinct
expression patterns in response to different stress treatments. To investigate
the in vivo function of FAO, transgenic Arabidopsis plants expressing sense,
antisense, and double-stranded BJFAO RNAs were generated and those with altered
activity of FAOs were selected for further characterization. It was found that
the shoot regeneration response in transgenic plants was significantly affected
by the modulated PA levels corresponding to FAO activities. Tissues that
originated from transgenic plants with down-regulated FAO activity were highly
regenerative, while those from transgenic plants with upregulated FAO activity
were poorly regenerative. The shoot regeneration capacity in these transgenic
plants was related to the levels of individual PAs, suggesting that FAO affects
shoot regeneration by regulating cellular PAs. Furthermore, it was found that the
effect of FAO activity on shoot regeneration was exerted downstream of the
Enhancer of Shoot Regeneration (ESR1) gene, which may function in a branch of the
cytokinin signalling pathway.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17122409 [PubMed - indexed for MEDLINE]

959: Plant Mol Biol. 2007 Feb;63(3):351-64. Epub 2006 Nov 21.

A rice gene activation/knockout mutant resource for high throughput functional
genomics.

Hsing YI, Chern CG, Fan MJ, Lu PC, Chen KT, Lo SF, Sun PK, Ho SL, Lee KW, Wang
YC, Huang WL, Ko SS, Chen S, Chen JL, Chung CI, Lin YC, Hour AL, Wang YW, Chang
YC, Tsai MW, Lin YS, Chen YC, Yen HM, Li CP, Wey CK, Tseng CS, Lai MH, Huang SC,
Chen LJ, Yu SM.

Institute of Plant and Microbial Biology, Academia Sinica, Nankang, Taipei 115,
Taiwan, ROC.

Using transfer DNA (T-DNA) with functions of gene trap and gene knockout and
activation tagging, a mutant population containing 55,000 lines was generated.
Approximately 81% of this population carries 1-2 T-DNA copies per line, and the
retrotransposon Tos17 was mostly inactive in this population during tissue
culture. A total of 11,992 flanking sequence tags (FSTs) have been obtained and
assigned to the rice genome. T-DNA was preferentially ( approximately 80%)
integrated into genic regions. A total of 19,000 FSTs pooled from this and
another T-DNA tagged population were analyzed and compared with 18,000 FSTs from
a Tos17 tagged population. There was difference in preference for integrations
into genic, coding, and flanking regions, as well as repetitive sequences and
centromeric regions, between T-DNA and Tos17; however, T-DNA integration was more
evenly distributed in the rice genome than Tos17. Our T-DNA contains an enhancer
octamer next to the left border, expression of genes within genetics distances of
12.5 kb was enhanced. For example, the normal height of a severe dwarf mutant,
with its gibberellin 2-oxidase (GA2ox) gene being activated by T-DNA, was
restored upon GA treatment, indicating GA2ox was one of the key enzymes
regulating the endogenous level of GA. Our T-DNA also contains a promoterless GUS
gene next to the right border. GUS activity screening facilitated identification
of genes responsive to various stresses and those regulated temporally and
spatially in large scale with high frequency. Our mutant population offers a
highly valuable resource for high throughput rice functional analyses using both
forward and reverse genetic approaches.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17120135 [PubMed - indexed for MEDLINE]

960: FEMS Microbiol Ecol. 2006 Dec;58(3):404-13.

Fungal endophytes in potato roots studied by traditional isolation and
cultivation-independent DNA-based methods.

Gö¶tz M, Nirenberg H, Krause S, Wolters H, Draeger S, Buchner A, Lottmann J, Berg
G, Smalla K.

Institute for Plant Virology, Microbiology and Biosafety, Federal Biological
Research Centre for Agriculture and Forestry (BBA), Braunschweig, Germany.

The composition and relative abundance of endophytic fungi in roots of
field-grown transgenic T4-lysozyme producing potatoes and the parental line were
assessed by classical isolation from root segments and cultivation-independent
techniques to test the hypothesis that endophytic fungi are affected by
T4-lysozyme. Fungi were isolated from the majority of root segments of both lines
and at least 63 morphological groups were obtained with Verticillium dahliae,
Cylindrocarpon destructans, Colletotrichum coccodes and Plectosporium tabacinum
as the most frequently isolated species. Dominant bands in the fungal
fingerprints obtained by denaturing gradient gel electrophoresis analysis of 18S
rRNA gene fragments amplified from total community DNA corresponded to the
electrophoretic mobility of the 18S rRNA gene fragments of the three most
abundant fungal isolates, V. dahliae, C. destructans and Col. coccodes, but not
to P. tabacinum. The assignment of the bands to these isolates was confirmed for
V. dahliae and Col. coccodes by sequencing of clones. Verticillium dahliae was
the most abundant endophytic fungus in the roots of healthy potato plants.
Differences in the relative abundance of endophytic fungi colonizing the roots of
T4-lysozyme producing potatoes and the parental line could be detected by both
methods.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17117985 [PubMed - indexed for MEDLINE]

961: J Agric Food Chem. 2006 Nov 29;54(24):8984-94.

Applications of metabolomics in agriculture.

Dixon RA, Gang DR, Charlton AJ, Fiehn O, Kuiper HA, Reynolds TL, Tjeerdema RS,
Jeffery EH, German JB, Ridley WP, Seiber JN.

Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway,
Ardmore, OK 73401, USA.

Biological systems are exceedingly complex. The unraveling of the genome in
plants and humans revealed fewer than the anticipated number of genes. Therefore,
other processes such as the regulation of gene expression, the action of gene
products, and the metabolic networks resulting from catalytic proteins must make
fundamental contributions to the remarkable diversity inherent in living systems.
Metabolomics is a relatively new approach aimed at improved understanding of
these metabolic networks and the subsequent biochemical composition of plants and
other biological organisms. Analytical tools within metabolomics including mass
spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy can profile
the impact of time, stress, nutritional status, and environmental perturbation on
hundreds of metabolites simultaneously resulting in massive, complex data sets.
This information, in combination with transcriptomics and proteomics, has the
potential to generate a more complete picture of the composition of food and feed
products, to optimize crop trait development, and to enhance diet and health.
Selected presentations from an American Chemical Society symposium held in March
2005 have been assembled to highlight the emerging application of metabolomics in
agriculture.

Publication Types:
Review

PMID: 17117782 [PubMed - indexed for MEDLINE]

962: Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18450-7. Epub 2006 Nov 20.

The root endophytic fungus Piriformospora indica requires host cell death for
proliferation during mutualistic symbiosis with barley.

Deshmukh S, Hö¼ckelhoven R, Schö¤fer P, Imani J, Sharma M, Weiss M, Waller F, Kogel
KH.

Research Centre for Biosystems, Land Use, and Nutrition, Institute of
Phytopathology and Applied Zoology, Justus Liebig Universitö¤t, D-35392 Giessen,
Germany.

Fungi of the recently defined order Sebacinales (Basidiomycota) are involved in a
wide spectrum of mutualistic symbioses (including mycorrhizae) with various
plants, thereby exhibiting a unique potential for biocontrol strategies. The
axenically cultivable root endophyte Piriformospora indica is a model organism of
this fungal order. It is able to increase biomass and grain yield of crop plants.
In barley, the endophyte induces local and systemic resistance to fungal diseases
and to abiotic stress. To elucidate the lifestyle of P. indica, we analyzed its
symbiotic interaction and endophytic development in barley roots. We found that
fungal colonization increases with root tissue maturation. The root tip meristem
showed no colonization, and the elongation zone showed mainly intercellular
colonization. In contrast, the differentiation zone was heavily infested by
inter- and intracellular hyphae and intracellular chlamydospores. The majority of
hyphae were present in dead rhizodermal and cortical cells that became completely
filled with chlamydospores. In some cases, hyphae penetrated cells and built a
meshwork around plasmolyzed protoplasts, suggesting that the fungus either
actively kills cells or senses cells undergoing endogenous programmed cell death.
Seven days after inoculation, expression of barley BAX inhibitor-1 (HvBI-1), a
gene capable of inhibiting plant cell death, was attenuated. Consistently, fungal
proliferation was strongly inhibited in transgenic barley overexpressing
GFP-tagged HvBI-1, which shows that P. indica requires host cell death for
proliferation in differentiated barley roots. We suggest that the endophyte
interferes with the host cell death program to form a mutualistic interaction
with plants.

PMID: 17116870 [PubMed - indexed for MEDLINE]

963: J Biotechnol. 2007 Feb 1;128(2):335-43. Epub 2006 Oct 27.

Improving the nutritive value of tubers: elevation of cysteine and glutathione
contents in the potato cultivar White Lady by marker-free transformation.

Stiller I, Dancs G, Hesse H, Hoefgen R, Bö¡nfalvi Z.

Agricultural Biotechnology Center, H-2101 Gö¶dö¶llo, Hungary.

The amino acids that limit the nutritive value of potato are the sulfur
containing amino acids methionine and cysteine. Manipulation of the targeted
amino acid biosynthesis is a way to circumvent this problem. Cysteine is
synthesised from O-acetyl-l-serine formed by serine acetyltransferase (SAT). To
increase the cysteine content of the commercial potato cultivar White Lady the
chimeric SAT-coding cysE gene from Escherichia coli under the control of the
constitutive CaMV 35S promoter and fused to the chloroplast targeting rbcS
5'-transit peptide sequence was introduced into the White Lady genome. Novelty of
the approach was the application of marker-free transformation. Two transgenic
lines were obtained that accumulated the cysE mRNA in high amounts. Crude leaf
extracts of these plants exhibited up to 80- and 20-fold higher SAT activity in
leaves and tubers, respectively, than those prepared from non-transformed plants.
Levels of cysteine and glutathione both in leaves and tubers were 1.5-fold higher
in average than in control plants. The alterations observed had no effect on
tuber yield and sprouting behaviour. Gas chromatography coupled to mass
spectrometry showed that all other amino acids than cysteine were unaffected.
Here we demonstrate for the first time that the cysteine content of tubers can be
enhanced by metabolic engineering.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17116342 [PubMed - indexed for MEDLINE]

964: FEBS J. 2006 Dec;273(23):5245-60.

Overexpression of putative topoisomerase 6 genes from rice confers stress
tolerance in transgenic Arabidopsis plants.

Jain M, Tyagi AK, Khurana JP.

Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular
Biology, University of Delhi South Campus, New Delhi, India.

DNA topoisomerase 6 (TOP6) belongs to a novel family of type II DNA
topoisomerases present, other than in archaebacteria, only in plants. Here we
report the isolation of full-length cDNAs encoding putative TOP6 subunits A and B
from rice (Oryza sativa ssp. indica), preserving all the structural domains
conserved among archaebacterial TOP6 homologs and eukaryotic meiotic
recombination factor SPO11. OsTOP6A1 was predominantly expressed in prepollinated
flowers. The transcript abundance of OsTOP6A2, OsTOP6A3 and OsTOP6B was also
higher in prepollinated flowers and callus. The expression of OsTOP6A2, OsTOP6A3
and OsTOP6B was differentially regulated by the plant hormones, auxin, cytokinin,
and abscisic acid. Yeast two-hybrid analysis revealed that the full-length
OsTOP6B protein interacts with both OsTOP6A2 and OsTOP6A3, but not with OsTOP6A1.
The nuclear localization of OsTOP6A3 and OsTOP6B was established by the transient
expression of their beta-glucuronidase fusion proteins in onion epidermal cells.
Overexpression of OsTOP6A3 and OsTOP6B in transgenic Arabidopsis plants conferred
reduced sensitivity to the stress hormone, abscisic acid, and tolerance to high
salinity and dehydration. Moreover, the stress tolerance coincided with enhanced
induction of many stress-responsive genes in transgenic Arabidopsis plants. In
addition, microarray analysis revealed that a large number of genes are expressed
differentially in transgenic plants. Taken together, our results demonstrate that
TOP6 genes play a crucial role in stress adaptation of plants by altering gene
expression.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17116242 [PubMed - indexed for MEDLINE]

965: Plant Physiol. 2007 Jan;143(1):225-35. Epub 2006 Nov 17.

Short vegetative phase-like MADS-box genes inhibit floral meristem identity in
barley.

Trevaskis B, Tadege M, Hemming MN, Peacock WJ, Dennis ES, Sheldon C.

Commonwealth Scientific and Industrial Research Organization, Division of Plant
Industry, Canberra, Australian Capitol Territory 2601, Australia.

Analysis of the functions of Short Vegetative Phase (SVP)-like MADS-box genes in
barley (Hordeum vulgare) indicated a role in determining meristem identity. Three
SVP-like genes are expressed in vegetative tissues of barley: Barley MADS1 (BM1),
BM10, and Vegetative to Reproductive Transition gene 2. These genes are induced
by cold but are repressed during floral development. Ectopic expression of BM1
inhibited spike development and caused floral reversion in barley, with florets
at the base of the spike replaced by tillers. Head emergence was delayed in
plants that ectopically express BM1, primarily by delayed development after the
floral transition, but expression levels of the barley VRN1 gene (HvVRN1) were
not affected. Ectopic expression of BM10 inhibited spike development and caused
partial floral reversion, where florets at the base of the spike were replaced by
inflorescence-like structures, but did not affect heading date. Floral reversion
occurred more frequently when BM1 and BM10 ectopic expression lines were grown in
short-day conditions. BM1 and BM10 also inhibited floral development and caused
floral reversion when expressed in Arabidopsis (Arabidopsis thaliana). We
conclude that SVP-like genes function to suppress floral meristem identity in
winter cereals.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17114273 [PubMed - indexed for MEDLINE]

966: Trends Biotechnol. 2007 Jan;25(1):2-6. Epub 2006 Nov 20.

EU biotech crop regulations and environmental risk: a case of the emperor's new
clothes?

Morris SH.

shane.morris@rogers.com

European Union Commissioner for the Environment Stavros Dimas recently hailed
'upgraded' non-genetically modified (GM) crops as an alternative to GM crops. A
comparative analysis of the environmental risks associated with such non-GM
herbicide-resistant crops and GM herbicide-resistant crops is presented here. The
analysis highlights serious weaknesses in the European Union (EU) regulatory
framework, and the contradictory policy of the EU Commission on the precautionary
principle is also shown. The continued political stance of ignoring these
regulatory and policy inconsistencies is examined and found to be flawed. It is
postulated that, even in the face of these flaws and coupled with recent
statements from the UK drawing attention to inconsistencies in the EU regulatory
framework, the EU will continue to ignore the real and present environmental
risks associated with upgraded non-GM crops for biopolitical reasons.

PMID: 17113665 [PubMed - indexed for MEDLINE]

967: FEBS Lett. 2006 Dec 11;580(28-29):6579-86. Epub 2006 Nov 13.

The mechanism of graft transmission of sense and antisense gene silencing in
tomato plants.

Shaharuddin NA, Han Y, Li H, Grierson D.

Plant Sciences Division, School of Biosciences, University of Nottingham, Sutton
Bonington Campus, Loughborough LE12 5RD, UK.

We investigated the effect of target mRNA level on grafting-transmitted gene
silencing in tomato plants by using a strong ACC oxidase 1 (ACO1) silencer as the
stock and transgenic ACO1 overexpressers as scions. Manifestation of graft
transmission of sense gene silencing required a high initial level of target mRNA
in the scion. A relatively high level of siRNA, similar to that in the strong
ACO1 silencer, was also detected in the silencing-susceptible strong ACO1
overexpressers prior to grafting. After grafting the silencing signal from the
stock enhanced the level of the siRNAs in the scion and the ACO1 mRNA level was
reduced dramatically. Using stock and scions producing different siRNAs we
provided evidence that the transmissible silencing signal does not correspond to
the bulk siRNAs in the stock. We also showed, contrary to a previous report, that
antisense silencing was graft-transmissible but it took longer to manifest
itself. The delay in graft transmission from antisense-silenced plants could be
attributed to the difference in the nature or strength of the signal or the
mechanism of its amplification, but is further evidence of mechanistic
similarities between sense and antisense silencing.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17113082 [PubMed - indexed for MEDLINE]

968: Plant Cell Rep. 2007 Apr;26(4):421-7. Epub 2006 Nov 17.

Compact shoot and leafy head 1, a mutation affects leaf initiation and
developmental transition in rice (Oryza sativa L).

Zhu QH, Dennis ES, Upadhyaya NM.

CSIRO Plant Industry, Australian Capital Territory, 2601, Canberra, Australia.
qianhao.zhu@csiro.au

The shoot apical meristem (SAM) produces lateral organs in a regular spacing
(phyllotaxy) and at a regular interval (phyllochron) during the vegetative phase.
In a Dissociation (Ds) insertion rice population, we identified a mutant, compact
shoot and leafy head 1 (csl1), which produced massive number of leaves (~70)
during the vegetative phase. In csl1, the transition from the vegetative to the
reproductive phase was delayed by about 2 months under long-day conditions. With
a reduced leaf size and severe dwarfism, csl1 failed to produce a normal panicle
after the transition to reproductive growth. Instead, it produced a leafy
panicle, in which all primary rachis-branches were converted to vegetative
shoots. Phenotypically csl1 resembled pla mutants in short plastochron but was
more severe in the conversion of the reproductive organs to vegetative organs. In
addition, neither the expression nor the coding region of PLA1 or PLA2 was
affected in csl1. csl1 is most likely a dominant mutation because no mutant
segregant was observed in progeny of 67 siblings of the csl1 mutant. CSL1 may
represent a novel gene, which functions downstream of PLA1 and/or PLA2, or
alternatively functions in a separate pathway, involved in the regulation of leaf
initiation and developmental transition via plant hormones or other mobile
signals.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17111113 [PubMed - indexed for MEDLINE]

969: Proc Natl Acad Sci U S A. 2006 Nov 28;103(48):18054-9. Epub 2006 Nov 16.

Engineering cottonseed for use in human nutrition by tissue-specific reduction of
toxic gossypol.

Sunilkumar G, Campbell LM, Puckhaber L, Stipanovic RD, Rathore KS.

Institute for Plant Genomics and Biotechnology and Department of Soil and Crop
Sciences, Texas A&M University, College Station, TX 77843, USA.

Global cottonseed production can potentially provide the protein requirements for
half a billion people per year; however, it is woefully underutilized because of
the presence of toxic gossypol within seed glands. Therefore, elimination of
gossypol from cottonseed has been a long-standing goal of geneticists. Attempts
were made to meet this objective by developing so-called "glandless cotton" in
the 1950s by conventional breeding techniques; however, the glandless varieties
were commercially unviable because of the increased susceptibility of the plant
to insect pests due to the systemic absence of glands that contain gossypol and
other protective terpenoids. Thus, the promise of cottonseed in contributing to
the food requirements of the burgeoning world population remained unfulfilled. We
have successfully used RNAi to disrupt gossypol biosynthesis in cottonseed tissue
by interfering with the expression of the delta-cadinene synthase gene during
seed development. We demonstrate that it is possible to significantly reduce
cottonseed-gossypol levels in a stable and heritable manner. Results from enzyme
activity and molecular analyses on developing transgenic embryos were consistent
with the observed phenotype in the mature seeds. Most relevant, the levels of
gossypol and related terpenoids in the foliage and floral parts were not
diminished, and thus their potential function in plant defense against insects
and diseases remained untouched. These results illustrate that a targeted genetic
modification, applied to an underutilized agricultural byproduct, provides a
mechanism to open up a new source of nutrition for hundreds of millions of
people.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17110445 [PubMed - indexed for MEDLINE]

970: Biotechnol J. 2006 Nov;1(11):1203-4.

BIOCENTURY, outrunner of China's biological technology in agriculture Biocentury
Transgene (China) Co., Ltd.

Cui H, Huang T.

cuihongzhi@biocentury.com.cn

PMID: 17109490 [PubMed - indexed for MEDLINE]

971: Reprod Nutr Dev. 2006 Sep-Oct;46(5):579-88. Epub 2006 Sep 23.

Preparation of recombinant proteins in milk to improve human and animal health.

Soler E, Thö©pot D, Rival-Gervier S, Jolivet G, Houdebine LM.

BioProtein Technologies 63, Domaine de Vilvert, 78350, Jouy-en-Josas, France.

Milk is a very abundant source of proteins for animal and human consumption. Milk
composition can be modified using transgenesis, including exogenous gene addition
and endogenous gene inactivation. The study of milk protein genes has provided
researchers with regulatory regions capable of efficiently and specifically
driving the expression of foreign genes in milk. The projects underway are aimed
at modifying milk composition, improving its nutritional value, reducing mammary
infections, providing consumers with antipathogen proteins and preparing purified
recombinant proteins for pharmaceutical use. The present paper summarises the
current progress in this field.

Publication Types:
Review

PMID: 17107647 [PubMed - indexed for MEDLINE]

972: Transgenic Res. 2007 Feb;16(1):51-63. Epub 2006 Nov 15.

Sources of uncertainty in the quantification of genetically modified oilseed rape
contamination in seed lots.

Begg GS, Cullen DW, Iannetta PP, Squire GR.

Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK.
gbegg@scri.ac.uk

Testing of seed and grain lots is essential in the enforcement of GM labelling
legislation and needs reliable procedures for which associated errors have been
identified and minimised. In this paper we consider the testing of oilseed rape
seed lots obtained from the harvest of a non-GM crop known to be contaminated by
volunteer plants from a GM herbicide tolerant variety. The objective was to
identify and quantify the error associated with the testing of these lots from
the initial sampling to completion of the real-time PCR assay with which the
level of GM contamination was quantified. The results showed that, under the
controlled conditions of a single laboratory, the error associated with the
real-time PCR assay to be negligible in comparison with sampling error, which was
exacerbated by heterogeneity in the distribution of GM seeds, most notably at a
small scale, i.e. 25 cm3. Sampling error was reduced by one to two thirds on the
application of appropriate homogenisation procedures.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17106767 [PubMed - indexed for MEDLINE]

973: Transgenic Res. 2007 Feb;16(1):121-6. Epub 2006 Nov 15.

Redirection of carotenoid metabolism for the efficient production of taxadiene
[taxa-4(5),11(12)-diene] in transgenic tomato fruit.

Kovacs K, Zhang L, Linforth RS, Whittaker B, Hayes CJ, Fray RG.

Plant Sciences Division, School of Biosciences, University of Nottingham, Sutton
Bonington Campus, Loughborough, LE12 5RD, UK.

The taxanes are a group of polycyclic diterpenes produced by various species of
yew. The potent anticancer drug paclitaxel (marketed as Taxol) is the
commercially most important taxane with annual sales in 2000 exceeding 1.6
billion dollars. Paclitaxel is currently obtained either by direct extraction
from yew trees or by the extraction of the precursor 10-deactilbaccatin III,
which is then converted to paclitaxel by semi-synthesis. Apart from cost, one of
the main draw backs to taxol in cancer treatment is the development of resistance
by tumours, commonly due to the expression of ABC transporter efflux pumps which
remove the drug from the target cell. A number of natural taxanes and
semisynthetic derivates, have recently been shown to act as potent inhibitors of
ABC transport proteins. These compounds have no effect upon microtubule
polymerization (the normal target of paclitaxel), but have the ability to restore
drug sensitivity when given in combination with paclitaxel to resistant cell
lines. In work to be described elsewhere, we sort to carry out a structure
function analysis of the ability of novel oxidised taxanes to act as ABC
transporter inhibitors. For this study 100 mg or more of taxadiene
[taxa-4(5),11(12)-diene], the first taxane in the paclitaxel pathway, was
required as starting material from which to synthesize these compounds. Taxadiene
is synthesised directly from geranylgeranyl diphosphate (GGPP), which is found in
most plant tissues where it serves as a common precursor for many metabolites.
The synthesis and use of GGDP are tightly regulated in most vegetative organs,
however, in tomato fruit it is used almost exclusively for the production of
coloured carotenoids which accumulate to high levels in the plastid as lycopene
crystals. Expressing taxadiene synthase in a yellow-fruited tomato line that
lacks the ability to utilise GGPP for carotenoid synthesis allowed GGPP normally
utilised for making carotenoids to be re-routed for the production of taxadiene,
allowing the facile extraction of 160 mg of highly pure taxadiene from 1 kg of
freeze dried fruit.

Publication Types:
In Vitro
Research Support, Non-U.S. Gov't

PMID: 17106766 [PubMed - indexed for MEDLINE]

974: Plant Cell Rep. 2007 Apr;26(4):467-77. Epub 2006 Nov 15.

Overexpression of barley hva1 gene in creeping bentgrass for improving drought
tolerance.

Fu D, Huang B, Xiao Y, Muthukrishnan S, Liang GH.

Department of Agronomy, Kansas State University, Manhattan, KS 66506, USA.

The objectives of this study were to test the feasibility of introducing barley
hva1 gene, a LEA3 member, into perennial grass species using the
Agrobacterium-mediated transformation technique and to determine whether
heterologous expression of hva1 would alleviate water-deficit injury in grass
species. Creeping bentgrass (Agrostis stolonifera var. palustris), a
drought-intolerant grass species, was transformed transiently or stably using
three different promoters in conjunction with the downstream report/target genes.
Two abscisic acid (ABA)-inducible promoters, ABA1 and ABA2 derived from
ABA-response complex (ABRC3) were used to examine stress-responsive expression of
the green fluorescent protein (GFP). Transient expression of GFP demonstrated the
inducibility of ABA1 and ABA2 promoters in response to exogenous ABA application.
The ABA2 promoter was further studied for stress-responsive expression of hva1
and a maize Ubi-1 promoter was tested for constitutive expression of the gene. In
the T(0) generation, the Ubi-1::hva1 transformants displayed variable expression
levels of HVA1 protein under normal growth conditions. The hva1 gene in the
ABA2::hva1 transformants maintained low expression under well-watered conditions,
but was upregulated under water-deficit conditions. The tolerance to water
deficit of T(0) transgenic lines was assessed by measuring leaf relative water
content and visually rating the severity of leaf wilting during to water stress.
Under water-stressed conditions, some transgenic lines maintained high water
content in leaves and showed significantly less extent of leaf wilting compared
with non-transgenic control plants. These results indicated that the introduction
of barley hva1 gene using constitutive or stress-inducible promoters lessened
water-deficit injury in creeping bentgrass, suggesting that heterologous
expression of LEA3 protein genes may enhance the survival ability of creeping
bentgrass in water limiting environments.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17106681 [PubMed - indexed for MEDLINE]

975: Transgenic Res. 2007 Jun;16(3):341-51. Epub 2006 Nov 14.

Transformation of rice with long DNA-segments consisting of random genomic DNA or
centromere-specific DNA.

Phan BH, Jin W, Topp CN, Zhong CX, Jiang J, Dawe RK, Parrott WA.

Department of Crop and Soil Sciences, University of Georgia, 111 Riverbend Road,
Athens, GA 30602, USA.

Rice was transformed with either long DNA-segments of random genomic DNA from
rice, or centromere-specific DNA sequences from either maize or rice. Despite the
repetitive nature of the transgenic DNA sequences, the centromere-specific
sequences were inserted largely intact and behave as simple Mendelian units.
Between 4 and 5% of bombarded callus clusters were transformed when bombarded
with just pCAMBIA 1305.2. Frequency of recovery dropped to 2-3% when BACs with
random genomic inserts were co-bombarded with pCAMBIA, and fell to less than 1%
when BACs with centromeric DNA inserts and pCAMBIA were co-bombarded. A similar
effect was noted on regeneration frequency. Differences in transformation
ability, regeneration and behavior of plants transgenic for BACs with random
genomic DNA inserts, as compared to those with centromeric DNA inserts, suggests
functional differences between these two types of DNA.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17103243 [PubMed - indexed for MEDLINE]

976: Transgenic Res. 2007 Jun;16(3):385-98. Epub 2006 Nov 14.

Production of siRNA targeted against TYLCV coat protein transcripts leads to
silencing of its expression and resistance to the virus.

Zrachya A, Kumar PP, Ramakrishnan U, Levy Y, Loyter A, Arazi T, Lapidot M, Gafni
Y.

Department of Genetics, Agricultural Research Organization, The Volcani Center,
P.O. Box 6, Bet Dagan 50250, Israel.

The coat protein (CP) of Tomato yellow leaf curl virus (TYLCV), encoded by the v1
gene, is the only known component of the viral capsid. In addition, the CP plays
a role in the virus transport into the host cell nucleus where viral genes are
replicated and transcribed. In this study, we analyzed the effect of small
interfering double-stranded RNAs (siRNAs), derived from an intron-hairpin RNA
(ihpRNA) construct and targeting the v1 gene product, on CP accumulation.
Transient assays involving agroinfiltration of the CP-silencing construct
followed by infiltration of a fused GFP-CP (green fluorescent protein-coat
protein) gene showed down-regulation of GFP expression in Nicotiana benthamiana.
Some of the transgenic tomato plants (cv. Micro-Tom), expressing the siRNA
targeted against the TYLCV CP gene, did not show disease symptoms 7 weeks
post-inoculation with the virus, while non-transgenic control plants were
infected within 2 weeks post inoculation. The present study demonstrates, for the
first time, that siRNA targeted against the CP of TYLCV can confer resistance to
the virus in transgenic tomato plants, thereby enabling flowering and fruit
production.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17103242 [PubMed - indexed for MEDLINE]

977: Transgenic Res. 2007 Aug;16(4):437-48. Epub 2006 Nov 14.

A leaf-based regeneration and transformation system for maize (Zea mays L.).

Ahmadabadi M, Ruf S, Bock R.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie, Am Mö¼hlenberg 1, D-14476,
Potsdam-Golm, Germany.

Efficient methods for in vitro propagation, regeneration, and transformation of
plants are of pivotal importance to both basic and applied research. While being
the world's major food crops, cereals are among the most difficult-to-handle
plants in tissue culture which severely limits genetic engineering approaches. In
maize, immature zygotic embryos provide the predominantly used material for
establishing regeneration-competent cell or callus cultures for genetic
transformation experiments. The procedures involved are demanding, laborious and
time consuming and depend on greenhouse facilities. We have developed a novel
tissue culture and plant regeneration system that uses maize leaf tissue and thus
is independent of zygotic embryos and greenhouse facilities. We report here: (i)
a protocol for the efficient induction of regeneration-competent callus from
maize leaves in the dark, (ii) a protocol for inducing highly regenerable callus
in the light, and (iii) the use of leaf-derived callus for the generation of
stably transformed maize plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17103238 [PubMed - indexed for MEDLINE]

978: Plant Cell Rep. 2007 Apr;26(4):439-47. Epub 2006 Nov 14.

Genetic improvement of the somatic embryogenesis and regeneration in soybean and
transformation of the improved breeding lines.

Kita Y, Nishizawa K, Takahashi M, Kitayama M, Ishimoto M.

Ehime Women's College, 421 Ibuki, Uwajima, Ehime, Japan.

Somatic embryos of soybean [Glycine max (L.) Merrill] have been used to generate
transgenic plants by particle bombardment. The induction and proliferation of
somatic embryos from immature cotyledons are dependent on the genotype of the
cultivar. Whereas somatic embryogenesis and plant regeneration are inefficient in
most cultivars, they are efficient in the cultivar Jack. We previously
established a breeding line, QF2, by the integration of null mutations of each
subunit of the major seed storage proteins glycinin and beta-conglycinin, but the
embryogenic response of this line is insufficient to allow efficient
transformation. We have now backcrossed QF2 to cultivar Jack in order to combine
the null traits with competence for somatic embryogenesis. The backcrossed
breeding lines selected on the basis of the absence of the major storage proteins
exhibited an improved capacity for the induction and proliferation of somatic
embryos compared with that of QF2. The induced somatic embryogenic tissue of
these breeding lines was successfully used for the production of transgenic
plants by particle bombardment. These results also indicate that somatic
embryogenesis in soybean is genetically controlled and inherited in a manner
independent of the null traits of the major seed storage proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17103216 [PubMed - indexed for MEDLINE]

979: Plant Cell Rep. 2007 Apr;26(4):459-65. Epub 2006 Nov 14.

Engineering resistance to PVY in different potato cultivars in a marker-free
transformation system using a 'shooter mutant' A. tumefaciens.

Bukovinszki A, Divö©ki Z, Csö¡nyi M, Palkovics L, Balö¡zs E.

Agricultural Biotechnology Center, Szent-Gyö¶rgyi A u 4, 2100, Gö¶dö¶llo, Hungary.
bagnes@abc.hu

In this work, Potato virus Y (PVY) resistant potatoes were generated using an
environmentally safe construct. For this purpose, a 'shooter' mutant
Agrobacterium-based transformation system was used. The isopentenyl transferase
gene (ipt) present on the Ti plasmid of 'shooter' strains enhances shoot
regeneration and can be used as a phenotypic selection marker. The introduced
marker-free binary vector carried a hairpin construct derived from the coat
protein gene of PVY-NTN strain in order to induce gene silencing. Transformation
resulted in high regeneration rates (1.4-5.7 shoots per explant). With
pre-selection for the ipt (+) phenotype the transformation frequency was 24-53%,
while without selection 12-28% of the shoots were PCR positive. The presence of
the transgene was verified by Southern hybridization. In 16 of 31 challenged
transformant lines PVY could be detected neither by RT-PCR nor by back
inoculation. A 62.5% of these resistant lines proved to be also ipt-free. This
transformation system was reproducible in four potato cultivars, suggesting that
it could easily be adapted for other species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17103215 [PubMed - indexed for MEDLINE]

980: Transgenic Res. 2007 Feb;16(1):109-13. Epub 2006 Nov 11.

Antioxidant value addition in human diets: genetic transformation of Brassica
juncea with gamma-TMT gene for increased alpha-tocopherol content.

Yusuf MA, Sarin NB.

School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India.

Alpha-tocopherol, the most biologically active form of vitamin E, is implicated
in decreasing the risk of several types of cancers, coronary heart disease and a
number of degenerative human conditions, when taken in excess of the recommended
daily allowance. Natural alpha-tocopherol has twice the bioavailability of the
synthetic isomer. This study describes a successful attempt at fortifying human
diets with natural alpha-tocopherol by taking recourse to genetic engineering of
an important oilseed crop, Brassica juncea. Gamma-tocopherol methyl transferase
cDNA from Arabidopsis thaliana, coding for the enzyme catalysing the conversion
of the large gamma-tocopherol pool to alpha-tocopherol, was overexpressed in B.
juncea plants. The successful integration of the transgene was confirmed by PCR
and Southern blot analysis, while the enhanced transcript level was evident in
the northern blot analysis. HPLC analysis of the seeds of the T1 transgenic lines
showed a shift in tocopherol profile with the highest over-expressors having
alpha-tocopherol levels as high as sixfold over the non-transgenic controls. This
study discusses the production of a transgenic oilseed crop with high
alpha-tocopherol levels, which can provide a feasible, innocuous, and inexpensive
way of taking the beneficial effects of high alpha-tocopherol intake to the
masses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17103027 [PubMed - indexed for MEDLINE]

981: Transgenic Res. 2007 Feb;16(1):85-97. Epub 2006 Nov 11.

Transgene-dependent incompatibility induced by introduction of the SK2:ZPT2-10
chimeric gene in petunia.

Kubo K, Takatsuji H.

Division of Plant Sciences, Plant Disease Resistance Research Unit, National
Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki,
305-8602, Japan.

In an attempt to functionally characterize a petunia zinc-finger gene ZPT2-10,
which is specifically expressed in style transmitting tissue, we fused its cDNA
downstream of the potato SK2 promoter (SK2:ZPT2-10) and then introduced it into
Petunia hybrida. We found that some transformants had acquired altered traits in
compatibility in mating; these were termed 'transgene-dependent incompatibility
(TDI)'. These transgenic lines were fertile when self-pollinated or crossed with
other TDI lines. In contrast, they failed to mate when crossed with untransformed
wild-type petunia or non-TDI lines of SK2:ZPT2-10 transformants. The TDI
phenomenon was observed irrespective of whether the TDI lines were used as the
pollen or pistillar parent. The TDI phenotype cosegregated with the SK2:ZPT2-10
transgene in the T1 generation and loss of this transgene resulted in the
recovery of normal fertility. In the case of infertile pollination with the TDI
line as one parent, pollen tubes grew normally through pistillar tissues, where
endogenous ZPT2-10 is expressed, and eventually reached the ovules. However, the
resultant embryos were arrested at the globular-heart stage. We found no
correlation between the occurrence of the TDI phenotype and the expression of
ZPT2-10 transcripts. On the basis of these observations, we discuss the possible
molecular mechanisms underlying this phenomena and its utility.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17103023 [PubMed - indexed for MEDLINE]

982: Plant Cell Rep. 2007 May;26(5):631-9. Epub 2006 Nov 11.

Co-bombardment, integration and expression of rice chitinase and thaumatin-like
protein genes in barley (Hordeum vulgare cv. Conlon).

Tobias DJ, Manoharan M, Pritsch C, Dahleen LS.

Department of Plant Sciences, North Dakota State University, Fargo, ND 58105,
USA.

Pathogenesis-related (PR) proteins associated with degradation of structural
components of pathogenic filamentous fungi were overexpressed in the two-rowed
malting barley (Hordeum vulgare L.) cultivar Conlon. Transgenes were introduced
by co-bombardment with two plasmids, one carrying a rice (Oryza sativa L.)
chitinase gene (chi11) and another carrying a rice thaumatin-like protein gene
(tlp). Each gene was under the control of the maize ubiquitin (Ubi1) promoter.
Fifty-eight primary transformants from three independent transformation events
were regenerated. T(1) plants with high rice chi11 and tlp protein expression
levels were advanced to identify T(2) homozygotes by herbicide spray and
subjected to further molecular analyses. T(3) progeny from one event (E2) had
stable integration and expression of the rice chi11 and tlp while those from the
other events (E1 and E3) showed stable integration only of tlp. The successful
production of these lines overexpressing the antifungal chi and tlp proteins
provides materials to test the effects of these genes on a variety of fungal
diseases that attack barley and to serve as potential additional sources of
disease resistance.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17103003 [PubMed - indexed for MEDLINE]

983: Plant Cell Rep. 2007 Apr;26(4):479-88. Epub 2006 Nov 11.

Overexpression of defense response genes in transgenic wheat enhances resistance
to Fusarium head blight.

Mackintosh CA, Lewis J, Radmer LE, Shin S, Heinen SJ, Smith LA, Wyckoff MN,
Dill-Macky R, Evans CK, Kravchenko S, Baldridge GD, Zeyen RJ, Muehlbauer GJ.

Department of Agronomy and Plant Genetics, University of Minnesota, 411 Borlaug
Hall, 1991 Upper Buford Circle, St Paul, MN 55108, USA.

Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other
Fusarium species, is a major disease problem for wheat production worldwide. To
combat this problem, large-scale breeding efforts have been established. Although
progress has been made through standard breeding approaches, the level of
resistance attained is insufficient to withstand epidemic conditions. Genetic
engineering provides an alternative approach to enhance the level of resistance.
Many defense response genes are induced in wheat during F. graminearum infection
and may play a role in reducing FHB. The objectives of this study were (1) to
develop transgenic wheat overexpressing the defense response genes
alpha-1-purothionin, thaumatin-like protein 1 (tlp-1), and beta-1,3-glucanase;
and (2) to test the resultant transgenic wheat lines against F. graminearum
infection under greenhouse and field conditions. Using the wheat cultivar
Bobwhite, we developed one, two, and four lines carrying the alpha-1-purothionin,
tlp-1, and beta-1,3-glucanase transgenes, respectively, that had statistically
significant reductions in FHB severity in greenhouse evaluations. We tested these
seven transgenic lines under field conditions for percent FHB disease severity,
deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels
(VSK). Six of the seven lines differed from the nontransgenic parental Bobwhite
line for at least one of the disease traits. A beta-1,3-glucanase transgenic line
had enhanced resistance, showing lower FHB severity, DON concentration, and
percent VSK compared to Bobwhite. Taken together, the results showed that
overexpression of defense response genes in wheat could enhance the FHB
resistance in both greenhouse and field conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17103001 [PubMed - indexed for MEDLINE]

984: Plant Physiol Biochem. 2006 Nov-Dec;44(11-12):766-75. Epub 2006 Nov 3.

Isolation and characterization of a novel potato Auxin/Indole-3-Acetic Acid
family member (StIAA2) that is involved in petiole hyponasty and shoot
morphogenesis.

Kloosterman B, Visser RG, Bachem CW.

Laboratory of Plant Breeding, Department of Plant Sciences, Graduate School of
Experimental Plant Sciences, Wageningen University and Research Center, P.O. Box
386, 6700 AJ Wageningen, The Netherlands. bjorn.kloosterman@wur.nl

Auxin/indole-3-acetid acid (Aux/IAA) proteins are short-lived transcriptional
regulators that mediate their response through interaction with auxin response
factors (ARF). Although 29 Aux/IAA proteins have been identified in Arabidopsis
thaliana, their individual functions are still poorly understood and are largely
defined by observed growth defects in gain-of-function mutant alleles. Here we
present the isolation and characterization of a novel Aux/IAA protein in potato
(Solanum tuberosum) that is named StIAA2. Down regulation of StIAA2 results in
distinctive phenotypes that include, increased plant height, petiole hyponasty
and extreme curvature of growing leaf primordia in the shoot apex. Gene
expression analysis of transgenic plants with reduced StIAA2 transcript levels
resulted in the identification of a number of genes with altered expression
profiles including another member of the Aux/IAA gene family (StIAA). The
phenotypes that were observed in the StIAA2 suppression clones can be associated
with both common as well as unique functional roles among Aux/IAA family members
indicating the importance of analyzing Aux/IAA expression in different plant
species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17098436 [PubMed - indexed for MEDLINE]

985: Theriogenology. 2007 Jan 1;67(1):188-97. Epub 2006 Nov 13.

Current status of regulating biotechnology-derived animals in Canada: animal
health and food safety considerations.

Kochhar HP, Evans BR.

Canadian Food Inspection Agency, Ottawa, Ont, Canada. hkochhar@inspection.gc.ca

Development of an effective regulatory system for genetically engineered animals
and their products has been the subject of increasing discussion among
researchers, industry and policy developers, as well as the public. Since
transgenesis and cloning are relatively new scientific techniques, transgenic
animals are 'novel' organisms for which there is limited information. The issues
associated with the regulation of transgenic animals pertain to environmental
impact, human food safety, animal health and welfare, trade and ethics. It is a
challenge for the developers to prove the safety of the products of
biotechnology-derived animals and also for regulators to regulate this
increasingly powerful technology with limited background information. In
principle, an effective regulatory sieve should permit safe products while
forming a formidable barrier for those posing an unacceptable risk. Regulatory
initiatives for biotechnology-derived animals and their products should be able
to ensure high standards for human and animal health, a sound scientific basis
for evaluation; transparency and public involvement, and maintenance of genetic
diversity. This review proposes a regulatory regime that is based on scientific
risk based assessment and approval of products or by-products of
biotechnology-derived animals and its application in context to Canadian
regulations.

Publication Types:
Review

PMID: 17097725 [PubMed - indexed for MEDLINE]

986: Food Chem Toxicol. 2007 Apr;45(4):551-62. Epub 2006 Oct 4.

Thirteen week feeding study with transgenic maize grain containing event
DAS-ö˜15ö˜7-1 in Sprague-Dawley rats.

MacKenzie SA, Lamb I, Schmidt J, Deege L, Morrisey MJ, Harper M, Layton RJ,
Prochaska LM, Sanders C, Locke M, Mattsson JL, Fuentes A, Delaney B.

DuPont Haskell Laboratory, Newark, DE, USA.

Maize line 1507, containing event DAS-ö˜15ö˜7-1 (1507), is a genetically modified
(GM) maize plant that expresses the cry1F gene from Bacillus thuringiensis (Bt)
sbsp. aizawai and the phosphinothricin-N-acetyltransferase (pat) gene from
Streptomyces viridochromogenes throughout the plant including in the grain
expression of the Cry1F protein confers in planta resistance to the European corn
borer (ECB; Ostrinia nubilalis Hö¼bner: Crambidae) and other lepidopteran pests.
Expression of the PAT protein confers tolerance to the herbicidal active
ingredient glufosinate-ammonium. The current study evaluated the nutritional
performance of rats fed diets containing 1507 maize grain in a subchronic rodent
feeding study. The grains in this study, 1507, its near-isogenic control (33P66),
and a non-GM commercial hybrid (33J56) contained similar amounts of proximates,
amino acids, minerals, anti-nutrients, and secondary metabolites. The subchronic
feeding study compared standard toxicology response variables in rats fed diets
containing 1507 maize grain with those in rats fed diets containing non-GM maize
grains. All diets were prepared according to the specifications of PMI Nutrition
International, LLC Certified Rodent LabDiet 5002 (PMI) 5002). Diets were fed ad
libitum to Sprague-Dawley rats for approximately 90 days. In-life response
variables included indicators of dietary performance and weekly evaluations for
clinical signs of toxicity. No toxicologically significant differences were
observed in the nutritional performance variables, clinical and neurobehavioral
signs, ophthalmology, clinical pathology (hematology, clinical chemistry,
coagulation, and urinalysis), organ weights, and gross and microscopic pathology
between any pair of treatment groups. These results demonstrate that 1507 maize
grain is as safe and as nutritious as non-GM maize grain.

PMID: 17097206 [PubMed - indexed for MEDLINE]

987: Transgenic Res. 2007 Feb;16(1):15-28. Epub 2006 Nov 10.

Agronomic performance and transcriptional analysis of carotenoid biosynthesis in
fruits of transgenic HighCaro and control tomato lines under field conditions.

Giorio G, Stigliani AL, D'Ambrosio C.

Metapontum Agrobios, S.S. Jonica 106, Km 448.2, Metaponto, MT , 75010, Italy.
ggiorio@agrobios.it

Genetic manipulation of carotenoid biosynthesis in higher plants has been the
objective of a number of biotechnology programs, e.g. the Golden Rice Program.
However, tomato (Solanum lycopersicum L.), which naturally accumulates lycopene
in fruits, has attracted the attention of many groups who have manipulated it to
increase or diversify carotenoid accumulation. One of the most significant
achievements was "HighCaro (HC)," a transgenic tomato plant constitutively
expressing the tomato lycopene beta-cyclase (tLcy-b), that produces orange fruits
due to the complete conversion of lycopene to beta-carotene. In this article we
report the results of a field trial conducted in Metaponto (Italy) on HC and on
two control genotypes to evaluate the stability of the transgenic trait and their
yield performances. Transcriptional regulation of eight genes involved in
carotenogenesis was assayed by quantitative real-time PCR (qRT-PCR) analysis on
fruits collected at four distinct development stages. Statistical analysis
results demonstrated that in field conditions the transgene maintained its
ability to induce the conversion of lycopene to beta-carotene. Moreover,
agronomic performances and fruit quality in the transgenic line were not impaired
by this metabolic disturbance. Results of qRT-PCR analysis suggested that
transcription of PSY-1, PDS and ZDS genes were developmentally regulated in both
genotypes. Unexpectedly, Lcy-b expression in transgenic fruits was also
developmentally regulated, despite the fact that the gene was driven by a
constitutive promoter. Our data provide evidence that in photosynthetic cells a
strict and aspecific mechanism controls the level of transcripts until the onset
of chromoplasts differentiation, at which point a gene-specific control on
transcription takes place.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17096211 [PubMed - indexed for MEDLINE]

988: Nat Biotechnol. 2006 Nov;24(11):1329; author reply 1331-3.

Comment on:
Nat Biotechnol. 2006 Jul;24(7):753.

'Cisgenic' as a product designation.

Giddings LV.

Publication Types:
Comment
Letter

PMID: 17093471 [PubMed - indexed for MEDLINE]

989: Nat Biotechnol. 2006 Nov;24(11):1329-31; author reply 1331-3.

Comment on:
Nat Biotechnol. 2006 Jul;24(7):753.

'Cisgenic' as a product designation.

de Cock Buning T, Lammerts van Bueren ET, Haring MA, de Vriend HC, Struik PC.

Publication Types:
Comment
Letter

PMID: 17093470 [PubMed - indexed for MEDLINE]

990: Nat Biotechnol. 2006 Nov;24(11):1327-9; author reply 1331-3.

Comment on:
Nat Biotechnol. 2006 Jul;24(7):753.

'Cisgenic' as a product designation.

Schubert D, Williams D.

Publication Types:
Comment
Letter

PMID: 17093469 [PubMed - indexed for MEDLINE]

991: Nat Biotechnol. 2006 Nov;24(11):1301-2.

Liberty Link rice raises specter of tightened regulations.

Vermij P.

Publication Types:
News

PMID: 17093456 [PubMed - indexed for MEDLINE]

992: Nature. 2006 Nov 9;444(7116):137.

A breed apart.

[No authors listed]

PMID: 17093424 [PubMed - indexed for MEDLINE]

993: Br J Nutr. 2006 Nov;96(5):820-9.

Transgenic maize in the presence of ampicillin modifies the metabolic profile and
microbial population structure of bovine rumen fluid in vitro.

Koch M, Strobel E, Tebbe CC, Heritage J, Breves G, Huber K.

Department of Physiology, School of Veterinary Medicine, Bischofsholer Damm
15/102, 30273 Hannover, Germany.

Recently, transgenic crops have been considered as possible donors of transgenes
that could be taken up by micro-organisms under appropriate conditions. In an in
vitro rumen simulation system, effects of ampicillin on microbial communities
growing either on rumen contents with transgenic maize carrying a gene that
confers resistance to ampicillin or its isogenic counterpart as substrates were
examined continuously over 13 d. Rate of production of SCFA was measured to
determine functional changes in the rumen model and single-strand conformational
polymorphism was used to detect alterations in structure of the microbial
community. Rumen contents treated with ampicillin displayed a marked decrease in
the rate of production of SCFA and diversity of the microbial community was
reduced severely. In the presence of transgenic maize, however, the patterns of
change of rumen micro-organisms and their metabolic profiles were different from
that of rumen fluid incorporating maize bred conventionally. Recovery of
propionate production was observed both in the rumen fluid fed transgenic and
conventional maize after a delay of several days but recovery occurred earlier in
fermenters fed transgenic maize. Alterations in the microbial population
structures resulting from the ampicillin challenge were not reversed during the
experimental run although there was evidence of adaptation of the microbial
communities over time in the presence of the antibiotic, showing that populations
with different microbial structures could resume a pre-challenge metabolic
profile following the introduction of ampicillin, irrespective of the source of
the plant material in the growth medium.

PMID: 17092369 [PubMed - indexed for MEDLINE]

994: Acta Biochim Biophys Sin (Shanghai). 2006 Nov;38(11):795-802.

Functional characterization of a putative nitrate transporter gene promoter from
rice.

Hu TZ, Cao KM, Xia M, Wang XP.

Department of Biochemistry, School of Life Science, Fudan University, Shanghai
200433, China.

Drought is one of the most significant abiotic stresses that influence plant
growth and development. Expression analysis revealed that OsNRT1.3, a putative
nitrate transporter gene in rice, was induced by drought. To confirm if the
OsNRT1.3 promoter can respond to drought stress, a 2019 bp upstream sequence of
OsNRT1.3 was cloned. Three OsNRT1.3 promoter fragments were generated by
5'-deletion, and fused to the beta-glucuronidase (GUS) gene. The chimeric genes
were introduced into rice plants. NRT2019::GUS, NRT1196::GUS and NRT719::GUS
showed similar expression patterns in seeds, roots, leaves and flowers in all
transgenic rice, and GUS activity conferred by different OsNRT1.3 promoter
fragments was significantly upregulated by drought stress, indicating that
OsNRT1.3 promoter responds to drought stress and the 719 bp upstream sequence of
OsNRT1.3 contains the drought response elements.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17091197 [PubMed - indexed for MEDLINE]

995: Zhong Xi Yi Jie He Xue Bao. 2006 Nov;4(6):639-43.

[Construction of a plant effective expression vector containing the gene of
hepatitis B virus surface antigen]

[Article in Chinese]

Lin BY, Jin ZQ, Li M.

Xiamen Overseas Chinese Subtropical Plant Introduction Garden, National Plant
Introduction Quarantine Base, Xiamen, Fujian Province 361002, China.

OBJECTIVE: To construct a plant effective expression vector driven by a fruit
specific promoter for the expression of hepatitis B virus surface antigen
(HBsAg), to further improve the expression of exogenous gene in plant, and to
prepare for the development of an effective anti-hepatitis vaccine. METHODS:
Tomato fruit-specific promoters' gene 2A12 and E8 were respectively introduced to
pBPFOmega7 to form pB2A12 and pBE8. The DNA fragment containing HBsAg-s gene from
plasmid YEP-HBs was inserted respectively into pB2A12 and pBE8 to form pB2A12-HBs
and pBE8-HBs. The fragment containing "p35S+2A12+Omega+HBsAg-s+Tnos" of the
pB2A12-HBs was sub-cloned into plasmid pCAMBIA1301 to yield the reconstructed
plant binary expression plasmid pCAM2A12-HBs, and the fragment containing
"p35S+E8+Omega+HBsAg-s+Tnos" of the pBE8-HBs was sub-cloned into plasmid
pCAMBIA1301 to yield the plasmid pCAME8-HBs. The inserted gene HBsAg and
fruit-specific promoters in the reconstructed plant binary expression vectors
were confirmed by sequencing. Then, pCAM2A12-HBs and pCAME8-HBs were directly
introduced into Agrobacterium tumefaciens strain EHA105. RESULTS: Digestion with
restriction enzymes proved that all recombinant vectors had the inserts with
expected length of the target fragments, and the sequencing results were
confirmed correct. CONCLUSION: In this study, plant expression vector containing
HBsAg gene driven by fruit specific promoter and CaMV35s promoter was
successfully constructed.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 17090384 [PubMed - indexed for MEDLINE]

996: J Allergy Clin Immunol. 2006 Nov;118(5):1176-83. Epub 2006 Sep 8.

Reduced allergenicity of tomato fruits harvested from Lyc e 1-silenced transgenic
tomato plants.

Le LQ, Mahler V, Lorenz Y, Scheurer S, Biemelt S, Vieths S, Sonnewald U.

Department of Biochemistry, Friedrich-Alexander University Erlangen-Nuremberg,
Erlangen, Germany.

BACKGROUND: Profilin is a small actin-binding protein that contributes to the
allergenic potency of many fruits and vegetables, including tomato. Two highly
similar genes encoding tomato profilin have been isolated and designated as
allergen Lyc e 1.01 and Lyc e 1.02. OBJECTIVE: The aim of the study was to
generate profilin-reduced hypoallergenic tomato fruits by silencing of both genes
in transgenic tomato plants by means of RNA interference (RNAi). METHODS: The
efficiency of gene silencing was documented by means of Northern blotting,
immunoblotting, and skin prick testing. RESULTS: Quantification of the remaining
protein revealed that profilin accumulation in transgenic fruits was decreased
10-fold compared with that seen in untransformed controls. This decrease was
sufficient to cause a reduced allergenic reactivity in patients with tomato
allergy, as determined with skin prick tests. Because most patients with tomato
allergy are not monosensitized to profilin, the IgE reactivity to the
profilin-silenced tomato fruits in vivo varied widely between individuals tested.
CONCLUSION: We could demonstrate the efficient silencing of both profilin genes
in transgenic tomato plants using RNAi. This resulted in Lyc e 1-diminished
tomato fruits, providing proof of concept and demonstrating that RNAi can be used
to design allergen-reduced food. However, simultaneous silencing of multiple
allergens will be required to design hypoallergenic tomatoes. CLINICAL
IMPLICATIONS: Our findings demonstrate the feasibility of creating low-allergenic
food by using RNAi. This concept constitutes a novel approach to allergen
avoidance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17088146 [PubMed - indexed for MEDLINE]

997: Planta. 2007 Apr;225(5):1277-85. Epub 2006 Nov 4.

Expression of Arabidopsis phytochelatin synthase in Indian mustard (Brassica
juncea) plants enhances tolerance for Cd and Zn.

Gasic K, Korban SS.

Department of Natural Resources and Environmental Sciences, 310 ERML, University
of Illinois, 1201 W. Gregory, Urbana, IL 61801, USA.

Phytochelatins (PCs) are heavy metal binding peptides that play an important role
in sequestration and detoxification of heavy metals in plants. In this study, our
goal was to develop transgenic plants with increased tolerance for and
accumulation of heavy metals from soil by expressing an Arabidopsis thaliana
AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica
juncea L.). A 35S promoter fused to a FLAG-tagged AtPCS1 cDNA was expressed in
Indian mustard, and transgenic lines, designated pc lines, were evaluated for
tolerance to and accumulation of Cd and Zn. Transgenic plants with moderate
AtPCS1 expression levels showed significantly higher tolerance to Cd and Zn
stress, but accumulated significantly less Cd and Zn than wild type plants in
both shoot and root tissues. However, transgenic plants with highest expression
of the transgene did not exhibit enhanced Cd and Zn tolerance. Shoots of
Cd-treated pc plants had significantly higher levels of phytochelatins and thiols
than wild-type plants. Significantly lower concentrations of gluthatione in
Cd-treated shoot and root tissues of transgenic plants were observed. Moderate
expression levels of phytochelatin synthase improved the ability of Indian
mustard to tolerate certain levels of heavy metals, but at the same time did not
increase the accumulation potential for Cd and Zn.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17086401 [PubMed - indexed for MEDLINE]

998: J Vet Med Sci. 2006 Oct;68(10):1113-5.

Effects of feeding calves genetically modified corn bt11: a clinico-biochemical
study.

Shimada N, Murata H, Mikami O, Yoshioka M, Guruge KS, Yamanaka N, Nakajima Y,
Miyazaki S.

Safety Research Team, National Institute of Animal Health, Ibaraki, Japan.

Genetically modified corn Bt11 is insect-resistant and expresses Cry1Ab toxin, an
insecticidal protein, in kernels. Although Bt11 corn is considered safe based on
animal performance, there are no reports available on the clinico-biochemical
effects of feeding it to cattle. In this study, we evaluated the effects of
feeding Bt11 to calves, using blood and ruminal clinico-biochemical parameters.
Our three-month-long feeding experiment demonstrated that calves (n=6), fed with
a ration containing 43.3% of Bt11 corn kernels as dry matter, did not develop any
discernible clinical, hematological, biochemical, or ruminal abnormalities as
compared with control calves (n=6) fed non-Bt11 corn. The results suggest that
the transgenic Bt11 has no negative clinico-biochemical effects on calves.

Publication Types:
Randomized Controlled Trial
Research Support, Non-U.S. Gov't

PMID: 17085894 [PubMed - indexed for MEDLINE]

999: Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17331-6. Epub 2006 Nov 3.

Telomere-mediated chromosomal truncation in maize.

Yu W, Lamb JC, Han F, Birchler JA.

Division of Biological Sciences, University of Missouri, Columbia, MO 65211, USA.

Direct repeats of Arabidopsis telomeric sequence were constructed to test
telomere-mediated chromosomal truncation in maize. Two constructs with 2.6 kb of
telomeric sequence were used to transform maize immature embryos by
Agrobacterium-mediated transformation. One hundred seventy-six transgenic lines
were recovered in which 231 transgene loci were revealed by a FISH analysis. To
analyze chromosomal truncations that result in transgenes located near
chromosomal termini, Southern hybridization analyses were performed. A pattern of
smear in truncated lines was seen as compared with discrete bands for internal
integrations, because telomeres in different cells are elongated differently by
telomerase. When multiple restriction enzymes were used to map the transgene
positions, the size of the smears shifted in accordance with the locations of
restriction sites on the construct. This result demonstrated that the transgene
was present at the end of the chromosome immediately before the integrated
telomere sequence. Direct evidence for chromosomal truncation came from the
results of FISH karyotyping, which revealed broken chromosomes with transgene
signals at the ends. These results demonstrate that telomere-mediated chromosomal
truncation operates in plant species. This technology will be useful for
chromosomal engineering in maize as well as other plant species.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17085598 [PubMed - indexed for MEDLINE]

1000: Science. 2006 Nov 24;314(5803):1295-8. Epub 2006 Nov 2.

Comment in:
Science. 2006 Nov 24;314(5803):1252-3.

Localization of iron in Arabidopsis seed requires the vacuolar membrane
transporter VIT1.

Kim SA, Punshon T, Lanzirotti A, Li L, Alonso JM, Ecker JR, Kaplan J, Guerinot
ML.

Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.

Iron deficiency is a major human nutritional problem wherever plant-based diets
are common. Using synchrotron x-ray fluorescence microtomography to directly
visualize iron in Arabidopsis seeds, we show that iron is localized primarily to
the provascular strands of the embryo. This localization is completely abolished
when the vacuolar iron uptake transporter VIT1 is disrupted. Vacuolar iron
storage is also critical for seedling development because vit1-1 seedlings grow
poorly when iron is limiting. We have uncovered a fundamental aspect of seed
biology that will ultimately aid the development of nutrient-rich seed,
benefiting both human health and agricultural productivity.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17082420 [PubMed - indexed for MEDLINE]

1001: Plant Cell Environ. 2006 Jun;29(6):1122-37.

Tonoplast-located GmCLC1 and GmNHX1 from soybean enhance NaCl tolerance in
transgenic bright yellow (BY)-2 cells.

Li WY, Wong FL, Tsai SN, Phang TH, Shao G, Lam HM.

Department of Biology, The Chinese University of Hong Kong, Hong Kong Special
Administrative Region (SAR), China.

Genes encoding ion transporters that regulate ion homeostasis in soybean have not
been carefully investigated. Using degenerate primers, we cloned a putative
chloride channel gene (GmCLC1) and a putative Na+/H+ antiporter gene (GmNHX1)
from soybean. Confocal microscopic studies using yellow fluorescent fusion
proteins revealed that GmCLC1 and GmNHX1 were both localized on tonoplast. The
expressions of GmCLC1 and GmNHX1 were both induced by NaCl or dehydration stress
imposed by polyethylene glycol (PEG). Using mitochondrial integrity and cell
death as the damage indicators, a clear alleviation under NaCl stress (but not
PEG stress) was observed in both GmCLC1 and GmNHX1 transgenic cells. Using
fluorescent dye staining and quenching, respectively, a higher concentration of
chloride ion (Cl-) or sodium ion (Na+) was observed in isolated vacuoles in the
cells of GmCLC1 and of GmNHX1 transgenic lines. Our result suggested that these
vacuolar-located ion transporters function to sequester ions from cytoplasm into
vacuole to reduce its toxic effects.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17080938 [PubMed - indexed for MEDLINE]

1002: Biotechnol Lett. 2006 Dec;28(24):1983-91. Epub 2006 Nov 2.

Review: Genetically modified plants for the promotion of human health.

Yonekura-Sakakibara K, Saito K.

RIKEN Plant Science Center, 1-7-22, Suehiro, Tsurumi-ku, Yokohama, 230-0045,
Japan.

Plants are attractive biological resources because of their ability to produce a
huge variety of chemical compounds, and the familiarity of production in even the
most rural settings. Genetic engineering gives plants additional characteristics
and value for cultivation and post-harvest. Genetically modified (GM) plants of
the "first generation" were conferred with traits beneficial to producers,
whereas GM plants in subsequent "generations" are intended to provide beneficial
traits for consumers. Golden Rice is a promising example of a GM plant in the
second generation, and has overcome a number of obstacles for practical use.
Furthermore, consumer-acceptable plants with health-promoting properties that are
genetically modified using native genes are being developed. The emerging
technology of metabolomics will also support the commercial realization of GM
plants by providing comprehensive analyzes of plant biochemical components.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17080241 [PubMed - indexed for MEDLINE]

1003: Protein Expr Purif. 2007 Feb;51(2):293-302. Epub 2006 Sep 16.

Production and characterization of human CTLA4Ig expressed in transgenic rice
cell suspension cultures.

Lee SJ, Park CI, Park MY, Jung HS, Ryu WS, Lim SM, Tan HK, Kwon TH, Yang MS, Kim
DI.

Boryung Pharmaceutical Co. Ltd., Ansan 425-120, Republic of Korea.

Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4I g) fusion protein,
a novel immunosuppressive agent, was expressed in transgenic rice cell suspension
culture and its characteristics and in vitro activities were investigated. The
expression vector pMYN409 was constructed to express hCTLA4I g under the control
of rice alpha-amylase 3D (RAmy3D) promoter. Transgenic calli were prepared by
particle bombardment mediated transformation and were screened for hCTLA4I g
expression using ELISA. Under the induction condition by sugar starvation,
suspension-cultured rice cells secreted hCTLA4I g into the media up to 31.4 mg/L
in flask culture. The rice-derived hCTLA4Ig (hCTLA4IgP) was purified from the
culture media with affinity chromatography using protein A and compared with
CHO-derived hCTLA4Ig (hCTLA4IgM). Recombinant hCTLA4IgP has molecular weight of
approximately 50 kDa on SDS-PAGE under reducing condition, which is a little
different from that of hCTLA4IgM probably due to the difference of carbohydrate
chain structures. Purified hCTLA4IgP was biologically active and was confirmed to
suppress T-cell proliferation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17079164 [PubMed - indexed for MEDLINE]

1004: Med Law. 2006 Sep;25(3):491-502.

Biotechnology entrepreneurship and ethics: principles, paradigms, and products.

Kuszler PC.

University of Washington School of Law, William H Gates Hall, Seattle, USA.

Biotechnology, whether in the context of new drugs derived from DNA and genetic
technology, genetically modified food, or biologics making use of living cells,
raises ethical concerns at a variety of different levels. At the research level,
there is concern that the very nature of research is being subverted, rather than
enhanced, by entrepreneurship. This area of ethical concern has intensified in
the United States as a result of the conflicts of interests resulting from the
growing alliance between University academia and private industry in the research
enterprise. As we travel down the research path into development of a drug or
technology, ethical questions arise with respect to protecting human subjects and
society from danger and exploitation by researchers. As development gives way to
marketing and dissemination of a new product, government regulators are pressed
to get drugs and biologics through the regulatory pipeline into the market
faster, walking an ethical tightrope between speed and safety. As new
biotechnology products enter the market place, doctors and patients traverse yet
another tightrope, that between unknown risk and the promise of benefit. And
finally, patent protection is increasingly viewed as a unethical culprit in
keeping prices high and depriving the global poor from lifesaving drugs and
biologics. Bioethics has, to date, been largely a creation of Western research
and medicine. As such it is wholly inadequate to respond to the cascade of
ethical issues that flow from a vibrant biotechnology industry. And if
biotechnology is in its infancy, as most believe, it is crucial that scientists,
entrepreneurs and governments engage in dialogue about the ethical and societal
questions raised on the road of scientific progress.

PMID: 17078522 [PubMed - indexed for MEDLINE]

1005: Gig Sanit. 2006 Jul-Aug;(4):7-11.

[Hygienic characteristics of foodstuffs containing genetically modified
components]

[Article in Russian]

Beliaev EN, Ivanov AA, Fokin MV.

The paper analyzes the results of the investigations of raw foods, foodstuffs for
genetically modified components, conducted by the state sanitary and
epidemiological service of the Russian Federation during its current sanitary
inspection. The presented materials cover 2003-2004. The findings suggest that
there is a great deal of foods containing genetically modified sources on the
market and show the priority groups of foodstuffs and the distribution of these
foods on the territory of the Russian Federation.

Publication Types:
English Abstract

PMID: 17078283 [PubMed - indexed for MEDLINE]

1006: Proc Natl Acad Sci U S A. 2006 Nov 7;103(45):16672-6. Epub 2006 Oct 30.

Changing green leaf volatile biosynthesis in plants: an approach for improving
plant resistance against both herbivores and pathogens.

Shiojiri K, Kishimoto K, Ozawa R, Kugimiya S, Urashimo S, Arimura G, Horiuchi J,
Nishioka T, Matsui K, Takabayashi J.

Center for Ecological Research, Kyoto University, Otsu 520-2113, Japan.

Green leaf volatiles (GLVs) are commonly emitted by green plants, and their
production is drastically enhanced when they are under biotic stress. To clarify
the ecological function of naturally emitted GLVs, we studied the response of
Arabidopsis, whose GLV biosynthesis had been modified, when subjected to
herbivory or a pathogenic infection. There was a significant increase in GLV
production after herbivory by cabbage white butterfly larvae and pathogen (gray
mold) infection in hydroperoxide lyase (HPL) sense Arabidopsis compared with WT
controls. The HPL sense modification resulted in the plant being more attractive
to the parasitic wasp Cotesia glomerata, leading to higher mortality of the
herbivores. The HPL sense modification also resulted in greater inhibition of
growth of the fungus. By contrast, HPL antisense Arabidopsis produced fewer GLVs,
attracted fewer parasitoids, and was more susceptible to the pathogens than the
WT control. These data show that (i) one of the ecological functions of GLV
biosynthesis related to resistance against both herbivores and pathogens, and
(ii) the genetic modification of GLV biosynthesis could be a unique approach for
improving plant resistance against such biotic stresses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17075049 [PubMed - indexed for MEDLINE]

1007: Mol Plant Microbe Interact. 2006 Nov;19(11):1216-28.

The eight amino-acid differences within three leucine-rich repeats between Pi2
and Piz-t resistance proteins determine the resistance specificity to Magnaporthe
grisea.

Zhou B, Qu S, Liu G, Dolan M, Sakai H, Lu G, Bellizzi M, Wang GL.

Department of Plant Pathology, Ohio State University, Columbus, OH 43210, USA.

The rice blast resistance (R) genes Pi2 and Piz-t confer broad-spectrum
resistance against different sets of Magnaporthe grisea isolates. We first
identified the Pi2 gene using a map-based cloning strategy. The Pi2 gene is a
member of a gene cluster comprising nine gene members (named Nbs1-Pi2 to
Nbs9-Pi2) and encodes a protein with a nucleotide-binding site and leucine-rich
repeat (LRR) domain. Fine genetic mapping, molecular characterization of the Pi2
susceptible mutants, and complementation tests indicated that Nbs4-Pi2 is the Pi2
gene. The Piz-t gene, a Pi2 allele in the rice cultivar Toride 1, was isolated
based on the Pi2 sequence information. Complementation tests confirmed that the
family member Nbs4-Piz-t is Piz-t. Sequence comparison revealed that only eight
amino-acid changes, which are confined within three consecutive LRR,
differentiate Piz-t from Pi2. Of the eight variants, only one locates within the
xxLxLxx motif. A reciprocal exchange of the single amino acid between Pi2 and
Piz-t did not convert the resistance specificity to each other but, rather,
abolished the function of both resistance proteins. These results indicate that
the single amino acid in the xxLxLxx motif may be critical for maintaining the
recognition surface of Pi2 and Piz-t to their respective avirulence proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17073304 [PubMed - indexed for MEDLINE]

1008: Transgenic Res. 2006 Dec;15(6):729-37. Epub 2006 Oct 27.

Generation of marker- and backbone-free transgenic potatoes by site-specific
recombination and a bi-functional marker gene in a non-regular one-border
agrobacterium transformation vector.

Kondrö¡k M, van der Meer IM, Bö¡nfalvi Z.

Agricultural Biotechnology Center, P.O. Box 411, H-2101, Gö¶dö¶llo, Hungary.

A binary vector, designated PROGMO, was constructed to assess the potential of
the Zygosaccharomyces rouxii R/Rs recombination system for generating marker- and
backbone-free transgenic potato (Solanum tuberosum) plants with high transgene
expression and low copy number insertion. The PROGMO vector utilises a
constitutively expressed plant-adapted R recombinase and a codA-nptII
bi-functional, positive/negative selectable marker gene. It carries only the
right border (RB) of T-DNA and consequently the whole plasmid will be inserted as
one long T-DNA into the plant genome. The recognition sites (Rs) are located at
such positions that recombinase enzyme activity will recombine and delete both
the bi-functional marker genes as well as the backbone of the binary vector,
leaving only the gene of interest flanked by a copy of Rs and RB. Efficiency of
PROGMO transformation was tested by introduction of the GUS reporter gene into
potato. It was shown that after 21 days of positive selection and using 300
mgl(-1 )5-fluorocytosine for negative selection, 29% of regenerated shoots
carried only the GUS gene flanked by a copy of Rs and RB. The PROGMO vector
approach is simple and might be widely applicable for the production of marker-
and backbone-free transgenic plants of many crop species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17072563 [PubMed - indexed for MEDLINE]

1009: Transgenic Res. 2007 Feb;16(1):1-13. Epub 2006 Oct 27.

Effects of potato plants expressing a barley cystatin on the predatory bug
Podisus maculiventris via herbivorous prey feeding on the plant.

Alvarez-Alfageme F, Martö­nez M, Pascual-Ruiz S, Castaö±era P, Diaz I, Ortego F.

Departamento de Biologö­a de Plantas, Centro de Investigaciones Biolö¨gicas, CSIC,
Ramiro de Maeztu 9, 28040, Madrid, Spain.

The aim of this study was to assess the effects of potato plants expressing a
barley cystatin on a potentially cystatin-susceptible natural enemy by predation
on susceptible and non-susceptible preys feeding on the plant. We have focussed
on the impact of the variant HvCPI-1 C68 --> G, in which the only cysteine
residue was changed by a glycine, on the growth and digestive physiology of the
Colorado potato beetle (CPB), Leptinotarsa decemlineata, and the Egyptian cotton
leafworm (ECW), Spodoptera littoralis. Moreover, we have studied the
prey-mediated effects of the barley cystatin at the third trophic level, using
the predatory spined soldier bug (SSB), Podisus maculiventris, as a model.
Feeding trials conducted with CPB larvae reared on transgenic potato plants
expressing the C68 --> G variant resulted in significantly lower weight gains
compared to those fed on non-transformed (NT) plants. On the contrary, larger
weight gains were obtained when ECW larvae, that lack digestive cysteine
proteases, were reared on transgenic potato expressing the cystatin, as compared
to larvae fed on NT plants. No negative effects on survival and growth were
observed when SSB nymphs were exposed to HvCPI-1 C68 --> G by predation on either
CPB or ECW larvae reared on transgenic potato plants expressing the barley
cystatin, despite the fact that the inhibitor suppressed in vitro gut proteolysis
of the predatory bug. To investigate the physiological background, biochemical
analysis were carried out on guts of insects dissected at the end of the feeding
assays.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17072562 [PubMed - indexed for MEDLINE]

1010: Biotechnol Lett. 2006 Dec;28(24):2039-48. Epub 2006 Oct 28.

Characterization of human cytotoxic T lymphocyte-associated antigen
4-immunoglobulin (hCTLA4Ig) expressed in transgenic rice cell suspension
cultures.

Jung HS, Koo JK, Lee SJ, Park CI, Shin JY, Kim MH, Tan HK, Lim SM, Kim DI.

Boryung Central Research Institute, Boryung Pharmaceutical Co. Ltd., Ansan,
Kyungki-do, 425-120, Korea.

The avidity for CD80Ig/CD86Ig and the in vitro immunosuppressive effect of
recombinant human cytotoxic T lymphocyte-associated antigen 4-immunoglobulin,
produced by transgenic rice cell suspension cultures (hCTLA4Ig(P)) with
CHO-derived recombinant hCTLA4Ig (hCTLA4Ig(M)), were measured. Surface plasmon
resonance (SPR) was used for kinetic binding analysis: hCTLA4Ig(P) and
hCTLA4Ig(M) had higher avidity for CD80Ig/CD86Ig than for CD28Ig, and the avidity
for CD80Ig/CD86Ig was similar. hCTLA4Ig(P) and hCTLA4Ig(M) had similar in vitro
immunosuppressive activity against the expression of T cell-derived cytokines,
such as IL-2, IL-4, and IFN-gamma, but did not suppress the expression of
macrophage-derived cytokines, including TNF-alpha and IL-1beta, as well as NO.
Thus the immunosuppressive mechanism of hCTLA4Ig(P) is also T cell-specific and
it could therefore be used as an immunosuppressive agent with an equivalent
potency to that of hCTLA4Ig(M).

Publication Types:
Comparative Study
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17072529 [PubMed - indexed for MEDLINE]

1011: J Genet. 2006 Aug;85(2):157-60.

Tissue-specific histochemical localization of iron and ferritin gene expression
in transgenic indica rice Pusa Basmati (Oryza sativa L.).

Sivaprakash KR, Krishnan S, Datta SK, Parida AK.

Plant Molecular Biology Lab, M. S. Swaminathan Research Foundation, Taramani,
Chennai 600 113, India.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17072086 [PubMed - indexed for MEDLINE]

1012: Theriogenology. 2007 Jan 1;67(1):1-206. Epub 2006 Oct 27.

Proceedings of the IETS (International Embryo Transfer Society) Pre-Conference
Symposia, Kyoto, Japan, 6 January 2007.

[No authors listed]

Publication Types:
Congresses
Overall

PMID: 17069880 [PubMed - indexed for MEDLINE]

1013: Ecol Appl. 2006 Oct;16(5):1967-74.

Relative fitness of transgenic vs. non-transgenic maize x teosinte hybrids: a
field evaluation.

Guadagnuolo R, Clegg J, Ellstrand NC.

Laboratoire de Botanique Evolutive, Institut de Botanique, Universitö© de
Neuchö¢tel, Switzerland. roberto.guadagnuolo@unine.ch

Concern has been often expressed regarding the impact and persistence of
transgenes that enter wild populations via gene flow. The impact of a transgene
and its persistence are largely determined by the relative fitness of transgenic
hybrids and hybrid derivatives compared to non-transgenic plants. Nevertheless,
few studies have addressed this question experimentally in the field. Despite the
economic importance of maize, and the fact that it naturally hybridizes with the
teosinte taxon Zea mays ssp. mexicana, sometimes known as "chalco teosinte," the
question has received little experimental attention in this system. Using a
glyphosate-tolerant maize cultivar and chalco teosinte as parental lines, we
carried out a field experiment testing (1) the relative fitness of maize x
teosinte hybrids, compared to their parental taxa, as well as (2) the relative
fitness of transgenic hybrids compared to non-transgenic hybrids created from the
same parental stock. In order to evaluate the influence of the transgenic
construct in different genetic backgrounds, our study included transgenic and
non-transgenic pure maize progeny from the cultivar as well. We measured both
vegetative and reproductive parameters. Our results demonstrated that hybrids
have greater vigor and produced more seeds than the wild parent. However, in the
absence of selective pressure from glyphosate herbicide, we did not observe any
direct positive or negative impact of the transgene on the fitness or vigor of
either the hybrids or pure maize progeny. We discuss our results in terms of the
potential for spontaneous transgene flow and introgression from transgenic maize
into sympatric teosinte.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17069387 [PubMed - indexed for MEDLINE]

1014: Plant Physiol Biochem. 2006 Oct;44(10):543-50. Epub 2006 Sep 29.

Effects of the overexpression of a soybean cytosolic glutamine synthetase gene
(GS15) linked to organ-specific promoters on growth and nitrogen accumulation of
pea plants supplied with ammonium.

Fei H, Chaillou S, Hirel B, Polowick P, Mahon JD, Vessey JK.

Department of Plant Science, University of Manitoba, Winnipeg, MB, Canada R3T
2N2.

A soybean cytosolic glutamine synthetase gene (GS15) fused to a constitutive
promoter (CaMV 35S), a putative nodule-specific promoter (LBC(3)), or a putative
root-specific promoter (rolD) was transformed into Pisum sativum L. cv.
Greenfeast. Four lines with single copies (Lines 1, 7, 8 and 9) and four lines
with two copies each of GS15 (Lines 2, 4, 6 and 11) were compared to the
wild-type (WT) parental line for levels of cytosolic glutamine synthetase (GS1),
glutamine synthetase (GS) activity, N accumulation, N derived form the atmosphere
(NDFA), and biomass of plants grown on 0.0, 0.1, 1.0 or 10.0 mM NH(4)(+).
Enhanced levels of GS1 were detected in leaves of one of the two lines
transformed with the 35S-GS15 construct, and all three lines containing the
rolD-GS15 construct. All three lines containing the LBC(3)-GS15 construct had
increased levels of GS1 in nodules. Despite the increased levels of GS1 in many
transformants, only the roots of lines containing the rolD-GS15 construct
consistently demonstrated enhanced levels of GS activity (up to 12-fold).
Positive responses in plant N content, NDFA, and biomass were rare, but increases
in plant biomass and N content of up to 17% and 54%, respectively, occurred in
some of the rolD-GS15 lines at certain levels of ammonium. In general, GS15 copy
number did not seem to differentially affect phenotype of the transformants, and
transformants respond to ammonium concentrations in similar patterns to that
previously observed with nitrate. Despite the fact that the rolD-GS15
transformants consistently resulted in increased GS activity in roots and
resulted in some occurrences of increases in biomass and plant N content, the
lack of consistent positive growth effect across all transformants indicates that
the generalized overexpression of GS1 in tissues holds little potential for
positive growth responses in pea.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17067806 [PubMed - indexed for MEDLINE]

1015: Annu Rev Plant Biol. 2007;58:1-19.

From analysis of mutants to genetic engineering.

von Wettstein D.

Department of Crop and Soil Sciences, School of Molecular Biosciences and Center
for Integrated Biotechnology, Washington State University, Pullman, WA
99164-6420, USA. diter@wsu.edu

This chapter describes the research of developing transgenic barley for synthesis
of recombinant proteins with practical significance and of metabolic engineering
of proanthocyanidin-free barley. The results were obtained by graduate students,
postdoctoral researchers, and visiting scientists at the Carlsberg Laboratory
from 1972-1996 and during the past ten years at Washington State University. It
is written in appreciation of their enthusiasm, skill, and perseverance.

Publication Types:
Review

PMID: 17067283 [PubMed - indexed for MEDLINE]

1016: Prikl Biokhim Mikrobiol. 2006 Sep-Oct;42(5):593-8.

[Optimization of the protocol for constructing transgenic plants of the white
cabbage brassica oleracea var. capitata]

[Article in Russian]

Gribova TN, Kamionskaia AM, Skriabin KG.

The strain Agrobacterium tumefaciens GV3101, which contains the pBar vector
carrying the phosphinothricin acetyltransferase gene (bar) under the control of
the 35SCMoV promoter and NOS 3' terminator, was used for genetic transformation
of four white cabbage lines, Ges-3, Drv-2, Zmu 7, and Meg 2. The effect of
different concentrations and combinations of phytohormones was studied, which
allowed for choosing the cultivation conditions that provided a 63-78%
regeneration efficiency. It was demonstrated that concerted action by natural and
synthetic cytokinins is necessary for the lines studied. Overall, 26 transgenic
plants were obtained using the optimized protocol for agrobacterial
transformation. The transgenic nature of these plants was confirmed by PCR and
dot-blot hybridization.

Publication Types:
English Abstract

PMID: 17066961 [PubMed - indexed for MEDLINE]

1017: J Econ Entomol. 2006 Oct;99(5):1641-50.

Dispersal of newly eclosed European corn borer adults (Lepidoptera: Crambidae)
from corn into small-grain aggregation plots.

Reardon BJ, Sumerford DV, Sappington TW.

Corn Insects and Crop Genetics Research Unit, USDA-ARS, Genetics Laboratory, Iowa
State University, Ames, IA 50011, USA.

Genetically modified, insecticidal Bacillus thuringiensis (Bt) corn, Zea mays L.,
hybrids are used throughout the Corn Belt for European corn borer, Ostrinia
nubilalis (Hö¼bner) (Lepidoptera: Crambidae), control. To slow development of Bt
corn resistance, the Environmental Protection Agency requires growers to plant a
refuge. Determining the appropriate distance between a refuge and Bt corn, and
development of mitigation-remediation strategies such as mass releases of
susceptible moths, requires an understanding of adult dispersal and mating
behavior. However, much remains unknown about these behaviors. Because mating
often occurs in grass near cornfields where adult O. nubilalis aggregate, we
planted small-grain plots as aggregation sites in an attempt to retain
mass-released adults. The objectives of this study were to examine influences of
pheromone lure, plant density, and plant species on distributions of feral and
newly emerged, laboratory-reared O. nubilalis among small-grain aggregation
plots. Feral adults were collected in aggregation plots in relative abundance,
indicating that small-grain plots were acceptable aggregation sites. In contrast,
newly emerged adults that were released weekly as dye-marked pupae were rarely
found in aggregation plots, with approximately 150-1,500-fold fewer adults
captured than expected if all released adults had occupied the plots for > or = 1
d. The majority of newly emerged adults did not colonize the aggregation plots,
suggesting that recently eclosed adults leave their natal field and do not
colonize the first aggregation sites encountered. Plant species significantly
influenced adult distributions among aggregation plots. Mass releases of
laboratory-reared pupae in the field may not be a viable remediation tactic
because almost all of the newly emerged adults dispersed beyond 300 m of the
release point.

PMID: 17066794 [PubMed - indexed for MEDLINE]

1018: J Biotechnol. 2007 Jan 30;128(1):194-203. Epub 2006 Sep 23.

Effect of storage and processing on plasmid, yeast and plant genomic DNA
stability in juice from genetically modified oranges.

Weiss J, Ros-Chumillas M, Peö±a L, Egea-Cortines M.

Agricultural Science and Technology Department, Genetics, Universidad Politö©cnica
de Cartagena, 30203 Cartagena, Spain. julia.weiss@upct.es

Recombinant DNA technology is an important tool in the development of plant
varieties with new favourable features. There is strong opposition towards this
technology due to the potential risk of horizontal gene transfer between
genetically modified plant material and food-associated bacteria, especially if
genes for antibiotic resistance are involved. Since horizontal transfer
efficiency depends on size and length of homologous sequences, we investigated
the effect of conditions required for orange juice processing on the stability of
DNA from three different origins: plasmid DNA, yeast genomic DNA and endogenous
genomic DNA from transgenic sweet orange (C. sinensis L. Osb.). Acidic orange
juice matrix had a strong degrading effect on plasmid DNA which becomes apparent
in a conformation change from supercoiled structure to nicked, linear structure
within 5h of storage at 4 degrees C. Genomic yeast DNA was degraded during
exposure to acidic orange juice matrix within 4 days, and also the genomic DNA of
C. sinensis suffered degradation within 2 days of storage as indicated by
amplification results from transgene markers. Standard pasteurization procedures
affected DNA integrity depending on the method and time used. Our data show that
the current standard industrial procedures to pasteurize orange juice as well as
its acidic nature causes a strong degradation of both yeast and endogenous
genomic DNA below sizes reported to be suitable for horizontal gene transfer.

PMID: 17064805 [PubMed - indexed for MEDLINE]

1019: J Biol Chem. 2007 Jan 5;282(1):625-36. Epub 2006 Oct 24.

Maize pollen coat xylanase facilitates pollen tube penetration into silk during
sexual reproduction.

Suen DF, Huang AH.

Center for Plant Cell Biology, Department of Botany and Plant Sciences,
University of California, Riverside, California 92521, USA.

Cell wall hydrolases are well documented to be present on pollen, but their roles
on the stigma during sexual reproduction have not been previously demonstrated.
We explored the function of the tapetum-synthesized xylanase, ZmXYN1, on maize
(Zea mays L.) pollen. Transgenic lines (xyl-less) containing little or no
xylanase in the pollen coat were generated with use of an antisense construct of
the xylanase gene-coding region driven by the XYN1 gene promoter. Xyl-less and
wild-type plants had similar vegetative growth. Electron microscopy revealed no
appreciable morphological difference in anther cells and pollen between xyl-less
lines and the wild type, whereas immunofluorescence microscopy and biochemical
analyses indicated an absence of xylanase on xyl-less pollen. Xyl-less pollen
germinated as efficiently as wild-type pollen in vitro in a liquid medium but
less so on gel media of increasing solidity or on silk, which is indicative of
partial impaired water uptake. Once germinated in vitro or on silk, the xyl-less
and wild-type pollen tubes elongated at comparable rates. Tubes of germinated
xyl-less pollen on silk did not penetrate into the silk as efficiently as tubes
of wild-type pollen, and this lower efficiency could be overcome by the addition
of xylanase to the silk. For wild-type pollen, coat xylanase activity on oat
spelled xylan in vitro and tube penetration into silk were inhibited by xylose
but not glucose. The overall findings indicate that maize pollen coat xylanase
facilitates pollen tube penetration into silk via enzymatic xylan hydrolysis.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17062571 [PubMed - indexed for MEDLINE]

1020: J Agric Food Chem. 2006 Nov 1;54(22):8640-7.

Safety assessment of cre recombinase.

Hileman RE, Bonner HK, Kaempfe TA, Hammond BG, Glenn KC.

Monsanto Company, St. Louis, Missouri 63167, USA. ronald.e.hileman@monsanto.com

Cre recombinase, when used as a tool in agricultural biotechnology, can precisely
excise DNA sequences that may be useful in the introduction of a new trait but
are not needed in the commercial product. Although the cre genetic material would
not be present in the final product, the present studies were performed to assess
the safety of Cre recombinase to provide confirmatory evidence of the safe use of
Cre-lox technology in agricultural biotechnology. Cre recombinase shares no
relevant sequence similarity to known allergens or toxins. When Cre recombinase
was exposed to a pH 1.2 solution of simulated gastric fluid lacking pepsin, CD
spectroscopy showed that there was a loss of secondary structure and that the
protein was no longer active in a functional assay. Cre recombinase was degraded
rapidly when exposed to pepsin in a standardized gastric digestion model;
therefore, Cre recombinase would not survive the harsh gastric environment. When
orally administered to mice as an acute dosage of 53 mg/kg of body weight, no
treatment-related adverse findings were observed. These data support the
conclusion that human and animal dietary exposure to Cre recombinase pose no
known safety concerns; consistent with the fact that bacteriophage P1, the source
of the cre gene and expressed protein, is commonly encountered in the environment
and in normal enteric bacteria without reports of adverse consequences.

PMID: 17061845 [PubMed - indexed for MEDLINE]

1021: Plant J. 2006 Nov;48(4):581-91. Epub 2006 Oct 19.

Over-expression of OsAGAP, an ARF-GAP, interferes with auxin influx, vesicle
trafficking and root development.

Zhuang X, Jiang J, Li J, Ma Q, Xu Y, Xue Y, Xu Z, Chong K.

Key Laboratory of Photosynthesis and Molecular Environmental Physiology,
Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.

Development and organogenesis in both dicot and monocot plants are highly
dependent on polar auxin transport (PAT), which requires the proper asymmetric
localization of both auxin influx and efflux carriers. In the model dicot plant
Arabidopsis thaliana, the trafficking and localization of auxin efflux
facilitators such as PIN-FORMED1 (PIN1) are mediated by GNOM, a
guanine-nucleotide exchange factor (GEF) for the ADP-ribosylation factor (ARF)
family of small GTPases, but molecular regulators of the auxin influx
facilitators remain unknown. Here, we show that over-expression of OsAGAP, an
ARF-GTPase-activating protein (ARF-GAP) in rice, impaired PAT and interfered with
both primary and lateral root development. The lateral root phenotype could be
rescued by the membrane-permeable auxin 1-naphthyl acetic acid, but not by indole
3-acetic acid (IAA) or by 2,4-dichloro-phenoxyacetic acid, which require influx
facilitators to enter the cells. OsAGAP-over-expressing plants had alterations in
vesicle trafficking and localization of the presumptive A. thaliana auxin-influx
carrier AUX1, but not in the localization of the auxin efflux facilitators.
Together, our data suggest that OsAGAP has a specific role in regulating vesicle
trafficking pathways such as the auxin influx pathway, which in turn controls
auxin-dependent root growth in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17059407 [PubMed - indexed for MEDLINE]

1022: Ann N Y Acad Sci. 2006 Aug;1072:176-86.

Therapeutic drug delivery by genetically modified Lactococcus lactis.

Steidler L, Rottiers P.

Alimentary Pharmabiotic Centre, Transgenic Bacteriology, University College Cork,
Western Road, Cork, Ireland. l.steidler@ucc.ie

Food-grade bacteria have been consumed throughout history without associated
pathologies and are, therefore, absolutely safe to ingest. Unexpectedly,
Lactococcus lactis (L. lactis), known from cheese production, can be genetically
engineered to constantly secrete satisfactory amounts of bioactive cytokines.
Both of these features enabled the development of a new kind of topical delivery
system: topical and active delivery of therapeutic proteins by genetically
modified micro-organisms. The host organism's record inspired the development of
applications that target intestinal diseases. In a variety of mouse models,
chronic colon inflammation can be successfully treated with (interleukin)
IL-10-secreting L. lactis. Trefoil factor (TFF) producer strains have also been
shown to be very effective in the treatment of acute colitis. Such novel
therapeutic strains are textbook examples of genetically modified (GM) organisms.
There are legitimate concerns with regard to the deliberate release of GM
micro-organisms. On development of these applications, therefore, we have
engineered these bacteria in such a way that biological containment is
guaranteed. The essential gene thyA, encoding thymidylate synthase, has been
exchanged for IL-10. This makes the GM strain critically dependent on thymidine.
Lack of thymidine, for example, resulting from thymidine consumption by
thyA-deficient strains-will irreversibly lead to induced "thymidine-less death."
This accomplishment has created the possibility of using this strategy for
application in human medicine.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17057198 [PubMed - indexed for MEDLINE]

1023: Plant Physiol. 2006 Dec;142(4):1559-73. Epub 2006 Oct 20.

EARLY RESPONSIVE TO DEHYDRATION 15, a negative regulator of abscisic acid
responses in Arabidopsis.

Kariola T, Brader G, Helenius E, Li J, Heino P, Palva ET.

Viikki Biocenter, Department of Biological and Environmental Sciences, Division
of Genetics, University of Helsinki, FIN-00014, Helsinki, Finland.

EARLY RESPONSIVE TO DEHYDRATION 15 (ERD15) is rapidly induced in response to
various abiotic and biotic stress stimuli in Arabidopsis (Arabidopsis thaliana).
Modulation of ERD15 levels by overexpression or RNAi silencing altered the
responsiveness of the transgenic plants to the phytohormone abscisic acid (ABA).
Overexpression of ERD15 reduced the ABA sensitivity of Arabidopsis manifested in
decreased drought tolerance and in impaired ability of the plants to increase
their freezing tolerance in response to this hormone. In contrast, RNAi silencing
of ERD15 resulted in plants that were hypersensitive to ABA and showed improved
tolerance to both drought and freezing, as well as impaired seed germination in
the presence of ABA. The modulation of ERD15 levels not only affected abiotic
stress tolerance but also disease resistance: ERD15 overexpression plants showed
improved resistance to the bacterial necrotroph Erwinia carotovora subsp.
carotovora accompanied with enhanced induction of marker genes for systemic
acquired resistance. We propose that ERD15 is a novel mediator of stress-related
ABA signaling in Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17056758 [PubMed - indexed for MEDLINE]

1024: Plant Physiol. 2006 Dec;142(4):1469-79. Epub 2006 Oct 20.

Molecular interactions of arabinogalactan proteins with cortical microtubules and
F-actin in Bright Yellow-2 tobacco cultured cells.

Sardar HS, Yang J, Showalter AM.

Department of Environmental and Plant Biology, Ohio University, Athens, Ohio
45701-2979, USA.

Arabinogalactan proteins (AGPs), a superfamily of plant hydroxyproline-rich
glycoproteins, are present at cell surfaces. Although precise functions of AGPs
remain elusive, they are widely implicated in plant growth and development. A
well-characterized classical tomato (Lycopersicon esculentum) AGP containing a
glycosylphosphatidylinositol plasma membrane anchor sequence was used here to
elucidate functional roles of AGPs. Transgenic tobacco (Nicotiana tabacum) Bright
Yellow-2 (BY-2) cells stably expressing green fluorescent protein (GFP)-LeAGP-1
were plasmolysed and used to localize LeAGP-1 on the plasma membrane and in
Hechtian strands. Cytoskeleton disruptors and beta-Yariv reagent (which binds and
perturbs AGPs) were used to examine the role of LeAGP-1 as a candidate linker
protein between the plasma membrane and cytoskeleton. This study used a
two-pronged approach. First, BY-2 cells, either wild type or expressing
GFP-microtubule (MT)-binding domain, were treated with beta-Yariv reagent, and
effects on MTs and F-actin were observed. Second, BY-2 cells expressing
GFP-LeAGP-1 were treated with amiprophosmethyl and cytochalasin-D to disrupt MTs
and F-actin, and effects on LeAGP-1 localization were observed. beta-Yariv
treatment resulted in terminal cell bulging, puncta formation, and
depolymerization/disorganization of MTs, indicating a likely role for AGPs in
cortical MT organization. beta-Yariv treatment also resulted in the formation of
thicker actin filaments, indicating a role for AGPs in actin polymerization.
Similarly, amiprophosmethyl and cytochalasin-D treatments resulted in
relocalization of LeAGP-1 on Hechtian strands and indicate roles for MTs and
F-actin in AGP organization at the cell surface and in Hechtian strands.
Collectively, these studies indicate that glycosylphosphatidylinositol-anchored
AGPs function to link the plasma membrane to the cytoskeleton.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17056757 [PubMed - indexed for MEDLINE]

1025: Plant Cell Physiol. 2006 Dec;47(12):1591-602. Epub 2006 Oct 20.

Conservation and diversification of meristem maintenance mechanism in Oryza
sativa: Function of the FLORAL ORGAN NUMBER2 gene.

Suzaki T, Toriba T, Fujimoto M, Tsutsumi N, Kitano H, Hirano HY.

Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, 113-0033
Japan.

To elucidate the genetic mechanism that regulates meristem maintenance in
monocots, here we have examined the function of the gene FLORAL ORGAN NUMBER2
(FON2) in Oryza sativa (rice). Mutations in FON2 cause enlargement of the floral
meristem, resulting in an increase in the number of floral organs, although the
vegetative and inflorescence meristems are largely normal. Molecular cloning
reveals that FON2 encodes a small secreted protein, containing a CLE domain, that
is closely related to CLAVATA3 in Arabidopsis thaliana. FON2 transcripts are
localized at the apical region in all meristems in the aerial parts of rice
plants, showing an expression pattern similar to that of Arabidopsis CLV3.
Constitutive expression of FON2 causes a reduction in the number of floral organs
and flowers, suggesting that both the flower and inflorescence meristems are
reduced in size. This action of FON2 requires the function of FON1, an ortholog
of CLV1. Constitutive expression of FON2 also causes premature termination of the
shoot apical meristem in Arabidopsis, a phenotype similar to that caused by
constitutive expression of CLV3. Together with our previous study of FON1, these
results clearly indicate that the FON1-FON2 system in rice corresponds to the CLV
signaling system in Arabidopsis and suggest that the negative regulation of stem
cell identity by these systems may be principally conserved in a wide range of
plants within the Angiosperms. In addition, we propose a model of the genetic
regulation of meristem maintenance in rice that includes an alternative pathway
independent of FON2-FON1.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17056620 [PubMed - indexed for MEDLINE]

1026: Theriogenology. 2007 Jan 1;67(1):185-7. Epub 2006 Oct 20.

Regulatory considerations on transgenic livestock in Japan in relation to the
Cartagena protocol.

Yamanouchi K.

Nippon Institute for Biological Science, 9-2221-1 Shin-Machi, Ome, Tokyo, Japan.
yamanokazu@aol.com

In Japan, the development and application of living modified organisms (LMOs) are
regulated by law (conservation and sustainable use of biological diversity law).
Procedures are classed as type 1 for the use of LMOs where no preventive measures
against their dispersal into the environment are required and type 2 for the use
of LMOs where preventive measures are stipulated. Development and research on
transgenic livestock falls under the responsibility of the Ministry of Education,
Culture, Science, Sports and Technology. Field use of transgenic livestock is
controlled by the Ministry of Agriculture, Forestry and Fisheries. The author
describes risk assessment and management of transgenic livestock by both
ministries.

Publication Types:
Review

PMID: 17055568 [PubMed - indexed for MEDLINE]

1027: Food Chem Toxicol. 2007 Mar;45(3):364-77. Epub 2006 Sep 14.

Safety testing of GM-rice expressing PHA-E lectin using a new animal test design.

Poulsen M, Schrö¸der M, Wilcks A, Kroghsbo S, Lindecrona RH, Miller A, Frenzel T,
Danier J, Rychlik M, Shu Q, Emami K, Taylor M, Gatehouse A, Engel KH, Knudsen I.

Department of Toxicology and Risk Assessment, Danish Institute for Food and
Veterinary Research, Mö¸rkhö¸j Bygade 19, DK-2860 Sö¸borg, Denmark. mop@dfvf.dk

The 90-day animal study is the core study for the safety assessment of
genetically modified foods in the SAFOTEST project. The model compound tested in
the 90-day study was a rice variety expressing the kidney bean Phaseolus vulgaris
lectin agglutinin E-form (PHA-E lectin). Female Wistar rats were given a
nutritionally balanced purified diet with 60% parental rice, 60% PHA-E rice or
60% PHA-E rice spiked with 0.1% recombinant PHA-E lectin for 90 days. This
corresponded to a mean daily PHA-E lectin intake of approximately 0, 30 and
100mg/kg body weight for each group, respectively. The spiking was used to
increase the specificity and to demonstrate the sensitivity of the study. A range
of biological, biochemical, microbiological and pathological parameters were
examined and significant differences in weight of small intestine, stomach and
pancreas and plasma biochemistry were seen between groups. Included in this paper
are also data from the molecular characterisation and chemical analysis of the
PHA-E rice, from the construction and production of the PHA-E lectin, and from
the preceding 28-day in vivo study where the toxicity of the pure PHA-E lectin
was determined. In conclusion, the combined use of information from the
compositional analysis, the 28-day study and the characterisation of the PHA-E
rice and the PHA-E lectin has improved the design of the 90-day study. The
spiking procedure has facilitated the interpretation of the results of the study
and transferred it into a valuable tool for the future safety testing of
genetically modified foods.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17052831 [PubMed - indexed for MEDLINE]

1028: Food Chem Toxicol. 2007 Mar;45(3):350-63. Epub 2006 Sep 14.

A 90-day safety study in Wistar rats fed genetically modified rice expressing
snowdrop lectin Galanthus nivalis (GNA).

Poulsen M, Kroghsbo S, Schrö¸der M, Wilcks A, Jacobsen H, Miller A, Frenzel T,
Danier J, Rychlik M, Shu Q, Emami K, Sudhakar D, Gatehouse A, Engel KH, Knudsen
I.

Department of Toxicology and Risk Assessment, Danish Institute for Food and
Veterinary Research, Mö¸rkhö¸j Bygade 19, DK-2860 Sö¸borg, Denmark. mop@dfvf.dk

Genetically modified plants expressing insecticidal traits offer a new strategy
for crop protection, but at the same time present a challenge in terms of food
safety assessment. The present 90-day feeding study was designed to assess the
safety of a rice variety expressing the snowdrop Galanthus nivalis lectin (GNA
lectin), and forms part of a EU-funded project where the objective has been to
develop and validate sensitive and specific methods to assess the safety of
genetically modified foods. Male and female Wistar rats were given a purified
diet containing either 60% genetically modified or parental rice for 90 days.
This corresponds to a mean daily GNA lectin intake of approximately 58 and
67mg/kg body weight for males and females, respectively. Prior to the animal
study comprehensive analytical characterization of both rice materials was
performed. The chemical analyses showed a number of statistically significant
differences, with the majority being within the ranges reported in the
literature. In the animal study a range of clinical, biological, immunological,
microbiological and pathological parameters were examined. A number of
significant differences were seen between groups fed the two diets, but none of
them were considered to be adverse. In conclusion, the design of the present
animal study did not enable us to conclude on the safety of the GM food.
Additional group(s) where the expressed gene products have been spiked to the
diet should be included in order to be able to distinguish whether the observed
effects were due to the GNA lectin per se or to secondary changes in the GM rice.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17052828 [PubMed - indexed for MEDLINE]

1029: Theriogenology. 2007 Jan 1;67(1):166-77. Epub 2006 Oct 18.

Compositional analysis of dairy products derived from clones and cloned
transgenic cattle.

Laible G, Brophy B, Knighton D, Wells DN.

AgResearch, Ruakura Research Centre, Hamilton, New Zealand.
goetz.laible@agresearch.co.nz

Cloning technology is an emerging biotechnological tool that could provide
commercial opportunities for livestock agriculture. However, the process is very
inefficient and the molecular events underlying the technology are poorly
understood. The resulting uncertainties are causing concerns regarding the safety
of food products derived from cloned livestock. There are similar concerns for
livestock produced by biotechnologies which enable the purposeful introduction of
genetic modifications. To increase the knowledge about food products from animals
generated by these modern biotechnologies, we assessed compositional differences
associated with milk and cheese derived from cloned and transgenic cows. Based on
gross composition, fatty acid and amino acid profiles and mineral and vitamin
contents, milk produced by clones and conventional cattle were essentially
similar and consistent with reference values from dairy cows farmed in the same
region under similar conditions. Whereas colostrum produced by transgenic cows
with additional casein genes had similar IgG secretion levels and kinetics to
control cows, milk from the transgenic cows had a distinct yellow appearance, in
contrast to the white color of milk from control cows. Processing of milk into
cheese resulted in differences in the gross composition and amino acid profiles;
'transgenic' cheese had lower fat and higher salt contents and small but
characteristic differences in the amino acid profile compared to control cheese.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17052749 [PubMed - indexed for MEDLINE]

1030: J Exp Bot. 2006;57(14):3801-11. Epub 2006 Oct 18.

Transport and metabolism of raffinose family oligosaccharides in transgenic
potato.

Hannah MA, Zuther E, Buchel K, Heyer AG.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie, D-14424 Potsdam, Germany.

Raffinose family oligosaccharides (RFOs) are involved in the storage and
transport of carbon and serve as compatible solutes for protection against
abiotic stresses like drought or cold. RFOs are usually transported in plant
species that load sugars symplastically into the phloem. Loading probably occurs
by a polymer trapping mechanism which establishes a concentration gradient of
assimilates between the mesophyll and the vasculature. Transgenic approaches have
demonstrated phloem transport of small molecules produced in the companion cells
of apoplastic loading species, but these molecules have been non-native transport
substances to plants. In this study, transgenic potato plants with constitutive
or companion cell specific overexpression of galactinol synthase (GS) or GS plus
raffinose synthase (RS) are characterized, which together provide new insights
into the metabolism and transport of RFOs in plants. It is demonstrated that
raffinose and galactinol are both transported in the phloem and that, whilst the
effect of GS overexpression is promoter-independent, that of RS is dependent on
the promoter used. The presence of significant amounts of galactinol in the
phloem is shown and also that transgenic potato is unable to transport large
amounts of raffinose despite high RS expression and substrate concentrations.
These data indicate that there may be additional features of intermediary cells,
the specialized companion cells of RFO transporting plants, required for
significant RFO synthesis and transport that are currently not well-understood.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17050641 [PubMed - indexed for MEDLINE]

1031: Food Chem Toxicol. 2007 Mar;45(3):339-49. Epub 2006 Sep 8.

A 90-day safety study of genetically modified rice expressing Cry1Ab protein
(Bacillus thuringiensis toxin) in Wistar rats.

Schrö¸der M, Poulsen M, Wilcks A, Kroghsbo S, Miller A, Frenzel T, Danier J,
Rychlik M, Emami K, Gatehouse A, Shu Q, Engel KH, Altosaar I, Knudsen I.

Department of Toxicology and Risk Assessment, Danish Institute for Food and
Veterinary Research, Mö¸rkhö¸j Bygade 19, DK-2860 Sö¸borg, Denmark.

An animal model for safety assessment of genetically modified foods was tested as
part of the SAFOTEST project. In a 90-day feeding study on Wistar rats, the
transgenic KMD1 rice expressing Cry1Ab protein was compared to its non-transgenic
parental wild type, Xiushui 11. The KMD1 rice contained 15mg Bt toxin/kg and
based on the average feed consumption the daily intake was 0.54mg Bt toxin/kg
body weight. No adverse effects on animal behaviour or weight gain were observed
during the study. Blood samples collected one week prior to sacrifice were
analyzed and compared for standard haematological and biochemical parameters. A
few parameters were significantly different, but all within the normal reference
intervals for rats of this breed and age and not in relation to any other
findings, thus not considered treatment related. Upon sacrifice a large number of
organs were weighed, macroscopic and histopathological examinations were
performed with only minor changes to report. The aim of the study was to use a
known animal model in performance of safety assessment of a GM crop, in this case
KMD1 rice. The results show no adverse or toxic effects of KMD1 rice when tested
in the design used in this 90-day study. Nevertheless the experiences from this
study lead to the overall conclusion that safety assessment for unintended
effects of a GM crop cannot be done without additional test group(s).

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 17050059 [PubMed - indexed for MEDLINE]

1032: Vaccine. 2007 Jan 8;25(4):591-8. Epub 2006 Aug 28.

Expression of the fusion glycoprotein of Newcastle disease virus in transgenic
rice and its immunogenicity in mice.

Yang ZQ, Liu QQ, Pan ZM, Yu HX, Jiao XA.

College of Bioscience and Biotechnology and Jiangsu Key Laboratory of Crop
Genetics and Physiology, Yangzhou University, 12 East Wenhui Road, Yangzhou,
Jiangsu 225009, PR China.

Transgenic plant has become an attractive bioreactor to produce high-value
medical peptides and proteins in biomedical research. In present study, two
expression cassettes, pUNDVF and pGNDVF containing the fusion protein gene of
Newcastle disease virus (NDV F) under the control of maize ubiquitin (Ubi)
promoter or rice glutelin (Gt1) promoter, respectively, were constructed, and
introduced into rice (Oryzy sativa L.) by Agrobacterium-mediated transformation.
A total of 12 independent transgenic rice lines were regenerated, and the result
from PCR analysis indicated that the T-DNA region containing the NDV F chimeric
gene had been integrated into the genome of transgenic rice plants. ELISA and
Western-blot analyses revealed that the NDV F protein could be expressed and
accumulated in both leaf and seed tissue of several transgenic rice plants.
Moreover, the immunogenicity of expressed proteins was tested in a mouse model
and the results showed that specific antibodies were elicited in mice immunized
intraperitoneally with crude protein extracts from transgenic rice plants. It
implied the potential of using transgenic rice-based expression systems as
supplementary bioreactor for NDV engineering subunit vaccine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17049688 [PubMed - indexed for MEDLINE]

1033: Neuroscience. 2007 Jan 5;144(1):17-25. Epub 2006 Oct 13.

The contribution of endogenous opioids to food reward is dependent on sex and
background strain.

Hayward MD, Low MJ.

Center for the Study of Weight Regulation, Oregon Health and Science University,
3181 Southwest Sam Jackson Park Road, Portland, OR 97239, USA.
Michael.Hayward@Xenogen.com

Complex behaviors such as those associated with reward to unconditioned positive
reinforcers are polygenic processes. In studies using genetically modified mice
specific for the endogenous opioid systems an observed phenotype in a complex
behavior is likely to be dependent on interacting genes which, in inbred mouse
lines, influence that phenotype. To address this issue we examined operant
responding for palatable food reinforcers in mice lacking the expression of
beta-endorphin, enkephalin or both peptides congenic to two different genetic
backgrounds; C57BL/6J and DBA/2J. These two inbred strains were chosen because
their endogenous opioid states differ and they respond differently to exogenous
opioids in many behavioral assays. We found that wildtype and mutant C57BL/6J
mice acquired operant responding for food reinforcers faster than DBA/2J mice,
regardless of their opioid genotype. Although wildtype DBA/2J mice had a
significant deficit in acquisition of bar-pressing behavior to reach a
pre-established performance criterion, no subsequent deficit was observed under
two different schedules of reinforcement. Additionally, we found that mice
lacking enkephalin had decreased motivation to bar press for palatable food
reinforcers under a progressive ratio regardless of sex or background strain. In
contrast, the only subset of beta-endorphin-deficient mice that had decreased
motivation to bar press under a progressive ratio was males on the C57BL/6J
background. Of the two classical endogenous opioid peptides with preferential
activation of the mu opioid receptor, the knockout models would suggest that
enkephalins play a more consistent role than beta-endorphin in mediating the
motivation for food reward when tested under a progressive ratio.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 17049174 [PubMed - indexed for MEDLINE]

1034: J Biosci Bioeng. 2006 Sep;102(3):162-70.

Development of simple and efficient in Planta transformation method for wheat
(Triticum aestivum L.) using Agrobacterium tumefaciens.

Supartana P, Shimizu T, Nogawa M, Shioiri H, Nakajima T, Haramoto N, Nozue M,
Kojima M.

Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu
University, 3-15-1 Tokida, Ueda, Nagano, Japan.

Wheat (Triticum aestivum L. var. Shiranekomugi) seeds were soaked in water at 22
degrees C for 1 d. Thereafter, the embryo of the soaked seeds was inoculated with
Agrobacterium tumefaciens by piercing a region of the embryonic apical meristem
with a needle that had been dipped in an A. tumefaciens inoculum. The inoculated
seeds were incubated at 22 degrees C for 2 d and sterilized by cefotaxime
(Claforan) (1000 ppm water solution) treatment and then vernalized at 5 degrees C
for 25 d. Finally, the seedlings were grown to maturation (T(0) plants) and
allowed to pollinate naturally for seed setting (T(1) plants) in pots under
nonsterile condition. To examine the transformation by various means, four
different strains of A. tumefaciens were used for transformation. The following
five lines of evidence proved the transformation: altered phenotype and its
transmittance to the next generation, resistance of T(1) seed germination to
geneticin or hygromycin B, the detection of a transgene in T(1) plants by PCR
analysis and Southern hybridization and the rescue of the plasmid consisting of
the integrated T-DNA and flanking wheat genome DNA from T(1) plants. The
transformation efficiency of T(1) plants, which were transformed using different
A. tumefaciens strains, was estimated to be 33% by PCR analysis, 75% by Southern
hybridization and 40% by plasmid rescue.

PMID: 17046528 [PubMed - indexed for MEDLINE]

1035: Nutr Metab Cardiovasc Dis. 2007 Feb;17(2):74-81. Epub 2006 Oct 13.

Assessment of usual dietary intake in population studies of gene-diet
interaction.

Tucker KL.

Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, 711
Washington Street, Boston, MA 02111, USA. katherine.tucker@tufts.edu


AIMS: Dietary intake is a critical environmental exposure when considering the
effect of many genetic factors on disease risk. However, dietary intake is a
complex and changing measure that requires particular care in assessment. DATA
SYNTHESIS: Although weighed diet records can theoretically provide the most
accurate assessment of intake, they are usually not realistic in large population
studies due to heavy respondent burden, likelihood of poor compliance, and the
cost of data entry. Multiple 24-h dietary recalls can provide excellent detail,
allowing for diverse dietary practices, but they are costly and require multiple
contacts with participants. Food frequency questionnaires are the most
cost-effective tool for assessing usual intake, particularly for micronutrients
with high day-to-day variability. However, they have limitations for diverse
populations and recent studies have questioned their ability to measure
macronutrient intakes for assessing diet and disease relationships. CONCLUSION:
At the present time, food frequencies remain the most cost-effective tool for
large population studies. However, their limitations must be fully appreciated
and demonstration of validity for nutrients of concern in the populations under
study is essential. When macronutrients are of key interest, consideration should
be given to the use of multiple recalls. Records may be used only in educated and
compliant populations. Continued efforts to improve dietary assessment
methodology must be investigated.

Publication Types:
Review

PMID: 17046222 [PubMed - indexed for MEDLINE]

1036: Adv Chronic Kidney Dis. 2006 Oct;13(4):374-85.

Animal models of obesity-associated chronic kidney disease.

Mak RH, Kuo HJ, Cheung WW.

Division of Pediatric Nephrology, Oregon Health & Science University, Portland,
OR 97239, USA. makr@ohsu.edu

Dramatic advances in basic science have been made in the past 50 years on the
basis of animal models of obesity and type II diabetes. Positional-cloning
strategies in rodents with spontaneous obesity have enabled landmark scientific
breakthroughs and defined the molecular scaffolding for the regulation of energy
homeostasis. Recently, studies in the general population suggest that obesity is
an independent risk factor for chronic kidney disease. To date, most of the
animal studies that investigated chronic kidney disease associated with obesity
and type II diabetes have largely been descriptive. We aim to provide an overview
of animal models used to investigate the mechanisms of obesity-associated chronic
kidney disease. Our overview is not meant to be an exhaustive list of all animal
models in the literature on this subject, but rather to illustrate the
experimental approaches. Because of space limitations, we have chosen to
concentrate on rodent models. These animal models will provide excellent tools
for in vivo testing of molecular mechanisms. Further hypothesis-driven research
into the mechanism of chronic kidney disease and their progression by use of
these models will provide important insights necessary to develop therapeutic
strategies for this significant complication of the worldwide epidemic of obesity
and type II diabetes.

Publication Types:
Research Support, N.I.H., Extramural
Review

PMID: 17045223 [PubMed - indexed for MEDLINE]

1037: J AOAC Int. 2006 Sep-Oct;89(5):1347-52.

Interlaboratory transfer of a real-time polymerase chain reaction assay for
quantitative detection of genetically modified maize event TC-1507.

La Paz JL, Garcö­a-Muniz N, Nadal A, Esteve T, Puigdomö¨nech P, Pla M.

Institut de Biologia Molecular de Barcelona, Departament de Genö©tica Molecular,
Consorci CSIC-IRTA, Jordi Girona 18-26,Barcelona, Spain.

A real-time polymerase chain reaction (QPCR) assay was developed for quantitative
detection of a genetically modified (GM) maize event TC-1507 and modification to
conventional PCR for qualitative purposes. Sequences 5'-flanking TC-1507
full-length insert were characterized and showed multiple rearrangements
involving insert and maize chloroplast fragments. The event specificity of the
TC-1507 assays was based on the detection of transgene and plant rearranged
sequences found to 5' flank the insertion site. They were fully specific and
exhibited a limit of detection below 10 target copies, allowing consistent
detection of 0.1% GM levels. The QPCR was highly linear and efficient and proved
adequate for quantification of GM contents, aiming at the fulfillment of legal
requirements established in the European Union (i.e., compulsory labeling of
TC-1507 levels >0.9%). It satisfactorily determined TC-1507 contents on different
matrixes and was successfully transferred a different laboratory.

Publication Types:
Evaluation Studies
Multicenter Study
Validation Studies

PMID: 17042186 [PubMed - indexed for MEDLINE]

1038: Plant Physiol. 2006 Dec;142(4):1759-70. Epub 2006 Oct 13.

Nuclear magnetic resonance spectroscopy-based metabolite profiling of transgenic
tomato fruit engineered to accumulate spermidine and spermine reveals enhanced
anabolic and nitrogen-carbon interactions.

Mattoo AK, Sobolev AP, Neelam A, Goyal RK, Handa AK, Segre AL.

Henry A. Wallace Beltsville Agricultural Research Center, United States
Department of Agriculture, Agricultural Research Service, Sustainable
Agricultural Systems Laboratory, Beltsville, Maryland 20705-2350, USA.
mattooa@ba.ars.usda.gov

Polyamines are ubiquitous aliphatic amines that have been implicated in myriad
processes, but their precise biochemical roles are not fully understood. We have
carried out metabolite profiling analyses of transgenic tomato (Solanum
lycopersicum) fruit engineered to accumulate the higher polyamines spermidine
(Spd) and spermine (Spm) to bring an insight into the metabolic processes that
Spd/Spm regulate in plants. NMR spectroscopic analysis revealed distinct
metabolite trends in the transgenic and wild-type/azygous fruits ripened off the
vine. Distinct metabolites (glutamine, asparagine, choline, citrate, fumarate,
malate, and an unidentified compound A) accumulated in the red transgenic fruit,
while the levels of valine, aspartic acid, sucrose, and glucose were
significantly lower as compared to the control (wild-type and azygous) red fruit.
The levels of isoleucine, glucose, gamma-aminobutyrate, phenylalanine, and
fructose remained similar in the nontransgenic and transgenic fruits. Statistical
treatment of the metabolite variables distinguished the control fruits from the
transgenic fruit and provided credence to the pronounced, differential metabolite
profiles seen during ripening of the transgenic fruits. The pathways involved in
the nitrogen sensing/signaling and carbon metabolism seem preferentially
activated in the high Spd/Spm transgenics. The metabolite profiling analysis
suggests that Spd and Spm are perceived as nitrogenous metabolites by the fruit
cells, which in turn results in the stimulation of carbon sequestration. This is
seen manifested in higher respiratory activity and up-regulation of
phosphoenolpyruvate carboxylase and NADP-dependent isocitrate dehydrogenase
transcripts in the transgenic fruit compared to controls, indicating high
metabolic status of the transgenics even late in fruit ripening.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17041034 [PubMed - indexed for MEDLINE]

1039: Plant Physiol. 2006 Dec;142(4):1664-82. Epub 2006 Oct 13.

Heterologous expression and molecular and cellular characterization of CaPUB1
encoding a hot pepper U-Box E3 ubiquitin ligase homolog.

Cho SK, Chung HS, Ryu MY, Park MJ, Lee MM, Bahk YY, Kim J, Pai HS, Kim WT.

Department of Biology, College of Science, Yonsei University, Seoul 120-749,
Korea.

The U-box motif is a conserved domain found in the diverse isoforms of E3
ubiquitin ligase in eukaryotes. From water-stressed hot pepper (Capsicum annuum
L. cv Pukang) plants, we isolated C. annuum putative U-box protein 1 (CaPUB1),
which encodes a protein containing a single U-box motif in its N-terminal region.
In vitro ubiquitination and site-directed mutagenesis assays revealed that CaPUB1
possessed E3 ubiquitin ligase activity and that the U-box motif was indeed
essential for its enzyme activity. RNA gel-blot analysis showed that CaPUB1 mRNA
was induced rapidly by a broad spectrum of abiotic stresses, including drought,
high salinity, cold temperature, and mechanical wounding, but not in response to
ethylene, abscisic acid, or a bacterial pathogen, suggesting its role in the
early events in the abiotic-related defense response. Because transgenic work was
extremely difficult in hot pepper, in this study we overexpressed CaPUB1 in
Arabidopsis (Arabidopsis thaliana) to provide cellular information on the
function of this gene in the development and plant responses to abiotic stresses.
Transgenic Arabidopsis plants that constitutively expressed the CaPUB1 gene under
the control of the cauliflower mosaic virus 35S promoter had markedly longer
hypocotyls and roots and grew more rapidly than the wild type, leading to an
early bolting phenotype. Microscopic analysis showed that 35S::CaPUB1 roots had
increased numbers of small-sized cells, resulting in disordered, highly populated
cell layers in the cortex, endodermis, and stele. In addition,
CaPUB1-overexpressing plants displayed increased sensitivity to water stress and
mild salinity. These results indicate that CaPUB1 is functional in Arabidopsis
cells, thereby effectively altering cell and tissue growth and also the response
to abiotic stresses. Comparative proteomic analysis showed that the level of RPN6
protein, a non-ATPase subunit of the 26S proteasome complex, was significantly
reduced in 35SCaPUB1 seedlings as compared to the wild type. Pull-down and
ubiquitination assays demonstrated that RPN6 interacted physically with CaPUB1
and was ubiquitinated in a CaPUB1-dependent manner in vitro. Although the
physiological function of CaPUB1 is not yet clear, there are several
possibilities for its involvement in a subset of physiological responses to
counteract dehydration and high-salinity stresses in transgenic Arabidopsis
seedlings.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17041029 [PubMed - indexed for MEDLINE]

1040: Genes Genet Syst. 2006 Aug;81(4):255-63.

Erratum in:
Genes Genet Syst. 2007 Jun;82(3):271.

Complete assignment of structural genes involved in flavonoid biosynthesis
influencing bulb color to individual chromosomes of the shallot (Allium cepa L.).

Masuzaki S, Shigyo M, Yamauchi N.

The United Graduate School of Agricultural Sciences, Tottori University, Tottori,
Japan.

We analyzed Japanese bunching onion (Allium fistulosum L.) - shallot (Allium cepa
L. Aggregatum group) alien chromosome addition lines in order to assign the genes
involved in the flavonoid biosynthesis pathway to chromosomes of the shallot. Two
complete sets of alien monosomic additions (2n = 2x + 1 = 17) were used for
determining the chromosomal locations of several partial sequences of candidate
genes, CHS, CHI, F3H, DFR, and ANS via analyses of PCR-based markers. The results
of DNA marker analyses showed that the CHS-A, CHS-B, CHI, F3H, DFR, and ANS genes
should be assigned to chromosomes 2A, 4A, 3A, 3A, 7A, and 4A, respectively. HPLC
analyses of 14 A. fistulosum - shallot multiple alien additions (2n = 2x + 2 - 2x
+ 7 = 18 - 23) were conducted to identify the anthocyanin compounds produced in
the scaly leaves. A direct comparison between the genomic constitution and the
anthocyanin compositions of the multiple additions revealed that a 3GT gene for
glucosylation of anthocyanidin was located on 4A. Thus, we were able to assign
all structural genes involved in flavonoid biosynthesis influencing bulb color to
individual chromosomes of A. cepa.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17038797 [PubMed - indexed for MEDLINE]

1041: Curr Genet. 2006 Dec;50(6):405-16. Epub 2006 Oct 11.

RNA editing site recognition in heterologous plant mitochondria.

Choury D, Araya A.

Laboratoire de Rö©plication et Expression des Gö©nomes Eucaryotes et Rö©troviraux,
UMR 5097, Centre National de la Recherche Scientifique and Universitö© Victor,
Segalen Bordeaux II, 146, Bordeaux Cedex, France.

RNA editing is a process that modifies the information content of mitochondrial
messenger RNAs in flowering plants changing specific cytosine residues into
uridine. To gain insight into editing site recognition, we used electroporation
to introduce engineered wheat (Triticum aestivum) or potato (Solanum tuberosum)
mitochondrial cox2 genes, and an atp9-containing chimeric gene, into non-cognate
mitochondria, and observed the efficiency of editing in these contexts. Both
wheat and potato mitochondria were able to express "foreign" constructs, and
their products were properly spliced. Seventeen and twelve editing sites are
present in the coding regions of wheat and potato cox2 transcripts, respectively.
Eight are common to both plants, whereas nine are specific to wheat, and four to
potato. An analogous situation is found for the atp9 mRNA coding regions from
these species. We found that both mitochondria were able to recognize sites that
are already present as T at the genomic level, making RNA editing unnecessary for
that specific residue in the cognate organelle. Our results demonstrate that
non-cognate mitochondria are able to edit residues that are not edited in their
own transcripts, and support the hypothesis that the same trans-acting factor may
recognize several editing sites.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17033819 [PubMed - indexed for MEDLINE]

1042: Nat Biotechnol. 2006 Oct;24(10):1200-1.

Comment in:
Nat Biotechnol. 2007 Jun;25(6):623; author reply 624.

Comment on:
Nat Biotechnol. 2005 Apr;23(4):429-30.

Potential impact and cost-effectiveness of Golden Rice.

Stein AJ, Sachdev HP, Qaim M.

Publication Types:
Comment
Letter
Research Support, Non-U.S. Gov't

PMID: 17033649 [PubMed - indexed for MEDLINE]

1043: Nat Biotechnol. 2006 Oct;24(10):1191-3. Epub 2006 Oct 5.

Comment in:
Nat Biotechnol. 2007 Feb;25(2):166-7; discussion 167.

Comment on:
Nat Biotechnol. 2003 Jan;21(1):3-4. Nat Biotechnol. 2004 Feb;22(2):133.

Turning plants into protein factories.

Fox JL.

Publication Types:
Comment
News

PMID: 17033647 [PubMed - indexed for MEDLINE]

1044: Nat Biotechnol. 2006 Oct;24(10):1178.

Comment in:
Nat Biotechnol. 2007 Jan;25(1):33-4.

Parallel universes?

[No authors listed]

An EU Commissioner has a meeting of minds with an antibiotech agitator.

Publication Types:
Editorial

PMID: 17033639 [PubMed - indexed for MEDLINE]

1045: Nat Biotechnol. 2006 Oct;24(10):1177.

Comment in:
Nat Biotechnol. 2007 Feb;25(2):165; discussion 165-6.

Why silence is not an option.

[No authors listed]

GM products will continue to be marginalized in Europe as long as industry
remains silent.

Publication Types:
Editorial

PMID: 17033637 [PubMed - indexed for MEDLINE]

1046: J Exp Bot. 2006;57(14):3825-36. Epub 2006 Oct 10.

Differential expression within the LOX gene family in ripening kiwifruit.

Zhang B, Chen K, Bowen J, Allan A, Espley R, Karunairetnam S, Ferguson I.

Laboratory of Fruit Molecular Physiology and Biotechnology/The State Agriculture
Ministry Laboratory of Horticultural Plant Growth, Development and Biotechnology,
Zhejiang University, Huajiachi Campus, Hangzhou 310029, China.

Real-time quantitative PCR was used to study lipoxygenase (LOX) gene expression
patterns in kiwifruit (Actinidia deliciosa [A. Chev.] C.F. Liang et A.R. Ferguson
var. deliciosa cv. Hayward) during fruit ripening, and in response to ethylene
and low temperature during post-harvest storage. Six LOX genes were identified
and cloned from a kiwifruit EST database. All were expressed in vegetative
tissues and in the fruit. Expression of AdLox1 and AdLox5 increased markedly as
fruit developed to the climacteric stage and were up-regulated by ethylene
treatment, following a similar pattern to LOX enzyme activity. By contrast,
AdLox2, AdLox3, and AdLox4 transcripts were negatively associated with ethylene
accumulation, and ethylene application enhanced the decline in transcript levels.
Transcripts of AdLox6 declined with fruit ripening. The fruit showed no ripening
changes at low temperature, where transcripts of AdLox1 and AdLox6 were slightly
induced about 72 h after harvest, suggesting an adaptive response to low
temperature. Transient expression of the ethylene-responsive AdLox1 gene in
tobacco leaves led to significant degradation of chlorophyll and promoted tissue
senescence, whereas AdLox2 had no such effect. The results showed that the six
LOX genes were differentially regulated during kiwifruit ripening and senescence,
forming two groups, one active in ripening and responsive to ethylene and the
other more constitutively expressed. The possible roles of individual LOX
isoforms in kiwifruit are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17032731 [PubMed - indexed for MEDLINE]

1047: J Exp Bot. 2006;57(14):3737-46. Epub 2006 Oct 10.

Low copy number gene transfer and stable expression in a commercial wheat
cultivar via particle bombardment.

Yao Q, Cong L, Chang JL, Li KX, Yang GX, He GY.

China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory, Huazhong
University of Science and Technology (HUST), 430074 Luoyu Road 1037, Wuhan,
Hubei, People's Republic of China.

Two groups of linear gene constructs (gus and bar, and 1Ax1 and bar) lacking
vector backbone sequences were independently transferred into the elite wheat
(Triticum aestivum L.) variety EM12, and genetically stable transgenic plants
with low copy number transgene integration were recovered. Co-transformation
experiments were carried out in parallel using either circular whole plasmid(s)
or linear gene cassettes which were purified from the same plasmid by restrictive
digestion, each cassette consisting of a promoter, an open reading frame, and a
terminator. Six transgenic wheat lines transformed with 1Ax1 plus bar gene
cassettes, five lines with gus plus bar gene cassettes, three lines with p1Ax1
plus pAHC20, and two lines with pAHC25 were regenerated with transformation
frequencies of 0.6, 0.5, 0.3, and 0.2%, respectively. Southern blotting analysis
showed that there were 1-4 hybridizing bands in transgenic lines carrying gene
cassettes, of which most lines displayed single-copy transgene insertion.
Expression analyses showed that 50.5% of the T1 lines carrying gus plus bar gene
cassettes have the expression signals of two genes. SDS-PAGE analysis of the T1
generation revealed that 71% of herbicide-resistant plants carrying 1Ax1 plus bar
gene cassettes expressed the high molecular weight subunit 1Ax1 in the endosperm.
Gene cassettes were transmitted and segregated in the subsequent generations, in
simple Mendelian ratios. In addition, reverse transcription-polymerase chain
reaction (RT-PCR) results confirmed that 1Ax1 gene cassettes were expressed
specifically in the endosperm of the transgenic wheat plant. It is proposed that
gene transfer using multiple gene cassettes offers an efficient and rapid method
to obtain the single-copy transgenic wheat.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17032730 [PubMed - indexed for MEDLINE]

1048: J Plant Physiol. 2006 Nov;163(11):1179-84. Epub 2005 Nov 18.

Transgenic Arabidopsis plants expressing the rice dehydroascorbate reductase gene
are resistant to salt stress.

Ushimaru T, Nakagawa T, Fujioka Y, Daicho K, Naito M, Yamauchi Y, Nonaka H, Amako
K, Yamawaki K, Murata N.

Department of Biology and Geoscience, Faculty of Science, Shizuoka University,
836 Ohya, Shizuoka 422-8529, Japan. sbtushi@ipc.shizuoka.ac.jp

Vitamin C (l-ascorbate) is important for antioxidative and metabolic functions in
both plants and humans. Ascorbate itself is oxidized to dehydroascorbate during
the process of antioxidation, and dehydroascorbate reductase (DHAR, EC 1.8.5.1)
re-reduces the oxidized ascorbate. Therefore, this enzyme is assumed to be
critical for ascorbate recycling. Here we show that the expression of rice DHAR
in transgenic Arabidopsis thaliana enhanced resistance to salt stress. Salt
tolerance was remarkably improved despite slight increases in DHAR activity and
total ascorbate. This study provides direct evidence for the importance of DHAR
in salt tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17032619 [PubMed - indexed for MEDLINE]

1049: J Agric Food Chem. 2006 Oct 18;54(21):8086-92.

Extraction of recombinant dog gastric lipase from transgenic corn seed.

Zhong Q, Gu Z, Glatz CE.

Department of Chemical and Biological Engineering, Iowa State University, Ames,
IA 50011, USA. qzhong@utk.edu

Several approaches were examined for extracting the relatively hydrophobic
protein recombinant dog gastric lipase (rDGL) expressed in the endosperm of
transgenic corn seed. The first approach used minimal processing of the seed
before extraction (i.e. simple grinding of whole seed) followed by selective
extraction to eliminate 72% of contaminant proteins without compromising rDGL
recovery from the meal of whole grain. The second approach added defatting of the
whole grain meal to reduce the amount of detergent in the subsequent step for
extracting rDGL. The third approach incorporated dry-milling of the corn to
recover an endosperm rich fraction, followed by extraction of this fraction. The
dry milling strategy was most effective, resulting in recovery of 35 U rDGL/g of
corn seed (50 U/g of endosperm) with a specific activity of 9 U/mg compared to 22
U and 3 U/mg for the first strategy and 36 U and 3.7 U/mg for the second. The
reductions in host protein contamination and lower detergent levels of the
endosperm route should simplify downstream purification steps.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17032014 [PubMed - indexed for MEDLINE]

1050: J Agric Food Chem. 2006 Oct 18;54(21):8082-5.

High content of resveratrol in lettuce transformed with a stilbene synthase gene
of Parthenocissus henryana.

Liu S, Hu Y, Wang X, Zhong J, Lin Z.

College of Life Science, National Key Laboratory of Protein Engineering, and
Plant Genetic Engineering, Peking University, Beijing 100871, China.

Resveratrol (trans-3,5,4'-trihydroxystilbene) is a plant phytoalexin which has
positive effects on human health. Stilbene synthase (STS) is a key enzyme
involved in resveratrol biosynthesis. To construct a vector for STS expression in
lettuce plant, a cDNA-encoding STS of Parthenocissus henryana was fused to the
Cauliflower mosaic virus (CaMV) 35S promoter, and the bar gene was used as a
selective marker gene. To increase the expression of STS, the expression cassette
was flanked by MARs. In transgenic lettuce plants, an additional compound was
identified as resveratrol by HPLC and ESI-MS. Quantitative analysis showed that
the average content of resveratrol reached 56.40 +/- 5.52 microg/g leaf fresh
weight, which was comparable to the amount in grape skin. Anticancer assay in
HeLa cells revealed that apoptosis was induced by 200 microM of resveratrol
extracted from transgenic lettuce.

PMID: 17032013 [PubMed - indexed for MEDLINE]

1051: Plant Cell Rep. 2007 Apr;26(4):531-8. Epub 2006 Oct 10.

Transgenic rice plants expressing trichothecene 3-O-acetyltransferase show
resistance to the Fusarium phytotoxin deoxynivalenol.

Ohsato S, Ochiai-Fukuda T, Nishiuchi T, Takahashi-Ando N, Koizumi S, Hamamoto H,
Kudo T, Yamaguchi I, Kimura M.

Plant & Microbial Metabolic Engineering Research Unit and Laboratory for
Remediation Research, Discovery Research Institute and Plant Science Center,
RIKEN, 2-1 Hirosawa, Wako, Saitama, Japan.

Fusarium head blight (FHB) is a devastating disease of small grain cereal crops
caused by the necrotrophic pathogen Fusarium graminearum and Fusarium culmorum.
These fungi produce the trichothecene mycotoxin deoxynivalenol (DON) and its
derivatives, which enhance the disease development during their interactions with
host plants. For the self-protection, the trichothecene producer Fusarium species
have Tri101 encoding trichothecene 3-O-acetyltransferase. Although transgenic
expression of Tri101 significantly reduced inhibitory action of DON on tobacco
plants, there are several conflicting observations regarding the phytotoxicity of
3-acetyldeoxynivalenol (3-ADON) to cereal plants; 3-ADON was reported to be
highly phytotoxic to wheat at low concentrations. To examine whether cereal
plants show sufficient resistance to 3-ADON, we generated transgenic rice plants
with stable expression and inheritance of Tri101. While root growth of wild-type
rice plants was severely inhibited by DON in the medium, this fungal toxin was
not phytotoxic to the transgenic lines that showed trichothecene 3-O-acetylation
activity. This is the first report demonstrating the DON acetylase activity and
DON-resistant phenotype of cereal plants expressing the fungal gene.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17031651 [PubMed - indexed for MEDLINE]

1052: Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18444-9. Epub 2006 Oct 9.

Combinatorially selected defense peptides protect plant roots from pathogen
infection.

Fang ZD, Laskey JG, Huang S, Bilyeu KD, Morris RO, Schmidt FJ, English JT.

Division of Plant Sciences, University of Missouri, Columbia, MO 65211, USA.

Agricultural productivity and sustainability are continually challenged by
emerging and indigenous pathogens. Currently, many pathogens can be combatted
only with biocides or environmentally dangerous fumigants. Here, we report a
rapid and pathogen-specific strategy to reduce infection by organisms that target
plant roots. Combinatorially selected defense peptides, previously shown to
effect premature encystment of Phytophthora capsici zoospores, were fused to
maize cytokinin oxidase/dehydrogenase as a display scaffold. When expressed in
tomato roots, the peptide-scaffold constructs were secreted and accumulated to
sufficient concentrations in the rhizosphere to induce zoospore encystment and
thereby deter taxis to the root surface. Pathogen infection was significantly
inhibited in roots expressing bioactive peptides fused to the maize cytokinin
oxidase/dehydrogenase scaffold. This peptide-delivery technology is broadly
applicable for rapid development of plant defense attributes against plant
pathogens.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17030803 [PubMed - indexed for MEDLINE]

1053: Biotechnol Lett. 2006 Dec;28(23):1877-88. Epub 2006 Sep 22.

IgE binding to proteins from sesame and assessment of allergenicity: implications
for biotechnology?

Orruö±o E, Morgan MR.

Procter Department of Food Science, University of Leeds, Leeds, UK,
prceo@leeds.ac.uk

Successful prediction of the potential allergenicity of a protein may be a key
factor in the development of novel, genetically modified foods. The use of the
decision tree approach for the prediction of allergenicity is discussed. The
methods currently used for identifying allergenic proteins (including use of IgE
from patient sera for recognition of proteins) are reviewed. Finally, a specific
review of the literature concerning identification of allergens from sesame leads
to the conclusion that in the absence of validated animal models, identification
of allergenicity (and, consequently, prediction of allergenicity) may be
problematic.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 17028779 [PubMed - indexed for MEDLINE]

1054: Genetics. 2006 Dec;174(4):2061-70. Epub 2006 Oct 8.

Molecular analysis, cytogenetics and fertility of introgression lines from
transgenic wheat to Aegilops cylindrica host.

Schoenenberger N, Guadagnuolo R, Savova-Bianchi D, Kö¼pfer P, Felber F.

Laboratoire de Botanique Evolutive, Institut de Botanique, Universitö© de
Neuchö¢tel, 2007 Neuchö¢tel, Switzerland. nicola.schoenenberger@unine.ch

Natural hybridization and backcrossing between Aegilops cylindrica and Triticum
aestivum can lead to introgression of wheat DNA into the wild species. Hybrids
between Ae. cylindrica and wheat lines bearing herbicide resistance (bar),
reporter (gus), fungal disease resistance (kp4), and increased insect tolerance
(gna) transgenes were produced by pollination of emasculated Ae. cylindrica
plants. F1 hybrids were backcrossed to Ae. cylindrica under open-pollination
conditions, and first backcrosses were selfed using pollen bags. Female fertility
of F1 ranged from 0.03 to 0.6%. Eighteen percent of the sown BC1s germinated and
flowered. Chromosome numbers ranged from 30 to 84 and several of the plants bore
wheat-specific sequence-characterized amplified regions (SCARs) and the bar gene.
Self fertility in two BC1 plants was 0.16 and 5.21%, and the others were
completely self-sterile. Among 19 BC1S1 individuals one plant was transgenic, had
43 chromosomes, contained the bar gene, and survived glufosinate treatments. The
other BC1S1 plants had between 28 and 31 chromosomes, and several of them carried
SCARs specific to wheat A and D genomes. Fertility of these plants was higher
under open-pollination conditions than by selfing and did not necessarily
correlate with even or euploid chromosome number. Some individuals having
supernumerary wheat chromosomes recovered full fertility.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17028347 [PubMed - indexed for MEDLINE]

1055: Regul Toxicol Pharmacol. 2007 Feb;47(1):37-47. Epub 2006 Oct 5.

Compositional assessment of event DAS-59122-7 maize using substantial
equivalence.

Herman RA, Storer NP, Phillips AM, Prochaska LM, Windels P.

Dow AgroSciences LLC, Indianapolis, IN 46268, USA. raherman@dow.com

Event DAS-59122-7 (Herculex RW) maize (Zea mays L.) plants were transformed to
express the Cry34Ab1 and Cry35Ab1 binary insecticidal crystal proteins originally
isolated from Bacillus thuringiensis Berliner (Bt) strain PS149B1. These proteins
protect maize roots from attack by corn rootworms, Diabrotica spp. DAS-59122-7
maize also contains the pat gene, originally isolated from Streptomyces
viridochromogenes, which confers tolerance to glufosinate-ammonium herbicides
(e.g. Liberty). We assessed the composition of these transgenic plants (with and
without Liberty herbicide treatment), grown at a total of eight fields sites over
2 years, by applying the principle of substantial equivalence. Forage and grain
samples were analyzed for proximates, fiber and minerals, and grain was further
analyzed for amino acids, fatty acids, vitamins, secondary metabolites and
anti-nutrients. Data plots were prepared that allow for efficient investigation
of equivalency between event DAS-59122-7 maize and a non-transgenic near-isogenic
maize line grown contemporaneously. Results demonstrated that DAS-59122-7 maize
is equivalent to non-transgenic maize with respect to these important
constituents.

Publication Types:
Comparative Study

PMID: 17027131 [PubMed - indexed for MEDLINE]

1056: J Plant Physiol. 2007 May;164(5):655-64. Epub 2006 Oct 4.

Transgenic rice plants ectopically expressing AtBAK1 are semi-dwarfed and
hypersensitive to 24-epibrassinolide.

Wang L, Xu YY, Li J, Powell RA, Xu ZH, Chong K.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, The Chinese Academy of Sciences, Beijing 100093, People's
Republic of China.

Brassinosteroids (BRs) are endogenous plant hormones essential for plant growth
and development. Brassinosteroid insensitive1 (BRI1)-assocaiated receptor kinase
(BAK1) is one of the key components in the BR signal transduction pathway due to
its direct association with the BR receptor, BRI1. Although BRI1 and its
orthologs have been identified from both dicotyledonous and monocotyledonous
plants, less is known about BAK1 and its orthologs in higher plants other than
Arabidopsis. This article provides the first piece of evidence that AtBAK1 can
greatly affect growth and development of rice plants when ectopically expressed,
suggesting that rice may share similar BR perception mechanism via BRI1/BAK1
complex. Interestingly, transgenic rice plants displayed semi-dwarfism and
shortened primary roots. Physiological analysis and cell morphology assay
demonstrated that the observed phenotypes in transgenic plants were presumably
caused by hypersensitivity to endogenous levels of BRs, different from BR
insensitive and deficient rice mutants. Consistently, several known BR inducible
genes were also upregulated in transgenic rice plants, further suggesting that
BAK1 was able to affect BR signaling in rice. On the other hand, the transgenic
plants generated by overproducing AtBAK1 may potentially have agricultural
applications because the dwarfed phenotype is generally resistant to lodging,
while the fertility remains unaffected.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17027118 [PubMed - indexed for MEDLINE]

1057: Genetika. 2006 Aug;42(8):1083-8.

[In planta transformation of maize through inoculation of Agrobacterium into the
silks]

[Article in Russian]

Chumakov MI, Rozhok NA, Velikov VA, Tyrnov VS, Volokhina IV.

Integration of T-DNA into the maize genome as a result of treatment of silks with
Agrobacterium cells, containing activated vir genes, was demonstrated. In planta
treatment of maize (Zea mays L) was performed during flowering in field. Cell
suspension of Agrobacterium tumefaiciens strain GV3101(pTd33), carrying activated
vir genes, was applied onto the previously isolated silks, which were afterwards
pollinated with the pollen of the same cultivar. Integration of T-DNA into maize
genome was confirmed by PCR (the nptII and gus reporter genes) and hystochemical
staining of the seedling tissues, obtained from the transformed seeds.
Amplification of the nptII gene showed the presence of about 60.3% of
PCR-positive plants out of the total number of kanamycin-resistant seedlings
examined, or 6.8% of the total of number of seedlings.

Publication Types:
English Abstract

PMID: 17025158 [PubMed - indexed for MEDLINE]

1058: Plant Cell Rep. 2007 Mar;26(3):327-36. Epub 2006 Oct 6.

Delayed flowering time in Arabidopsis and Brassica rapa by the overexpression of
FLOWERING LOCUS C (FLC) homologs isolated from Chinese cabbage (Brassica rapa L.:
ssp. pekinensis).

Kim SY, Park BS, Kwon SJ, Kim J, Lim MH, Park YD, Kim DY, Suh SC, Jin YM, Ahn JH,
Lee YH.

National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea.

Chinese cabbage plants remain in the vegetative growth phase until they have
experienced prolonged exposure to cold temperature, known as vernalization. This
inhibition of flowering is caused by the high levels of FLOWERING LOCUS C (FLC)
expression. To increase the product value of Chinese cabbage by inhibiting the
floral transition, three genes (BrFLC1, BrFLC2, and BrFLC3) homologous to the
AtFLC gene, which encodes a floral repressor, were isolated from the Chinese
cabbage 'Chiifu'. These genes showed high similarity to AtFLC, although the
putative BrFLC1 protein contained ten more residues than AtFLC. The BrFLC genes
were expressed ubiquitously, except that BrFLC3 was not expressed in roots.
BrFLC1 and BrFLC2 showed stronger expression than BrFLC3 in unvernalized and
vernalized Chinese cabbage. The expression levels of the three BrFLC genes were
lower in an early-flowering Chinese cabbage, suggesting that the BrFLC transcript
level was associated with flowering time. Constitutive expression of the BrFLC
genes in Arabidopsis significantly delayed flowering, which was also observed in
transgenic Chinese cabbage overexpressing BrFLC3. These results suggest that the
BrFLC genes act similarly to AtFLC. Our results provide a technique for
controlling flowering time in Chinese cabbage and other crops to produce high
yields of vegetative tissues.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17024448 [PubMed - indexed for MEDLINE]

1059: EMBO J. 2006 Oct 18;25(20):4909-20. Epub 2006 Oct 5.

EBP1 regulates organ size through cell growth and proliferation in plants.

Horvö¡th BM, Magyar Z, Zhang Y, Hamburger AW, Bakö¨ L, Visser RG, Bachem CW, Bö¶gre
L.

Laboratory of Plant Breeding, Department of Plant Sciences, Graduate School of
Experimental Plant Sciences, Wageningen University and Research Centre,
Wageningen, The Netherlands. Beatrix.Horvath@WUR.nl

Plant organ size shows remarkable uniformity within species indicating strong
endogenous control. We have identified a plant growth regulatory gene,
functionally and structurally homologous to human EBP1. Plant EBP1 levels are
tightly regulated; gene expression is highest in developing organs and correlates
with genes involved in ribosome biogenesis and function. EBP1 protein is
stabilised by auxin. Elevating or decreasing EBP1 levels in transgenic plants
results in a dose-dependent increase or reduction in organ growth, respectively.
During early stages of organ development, EBP1 promotes cell proliferation,
influences cell-size threshold for division and shortens the period of
meristematic activity. In postmitotic cells, it enhances cell expansion. EBP1 is
required for expression of cell cycle genes; CyclinD3;1, ribonucleotide reductase
2 and the cyclin-dependent kinase B1;1. The regulation of these genes by EBP1 is
dose and auxin dependent and might rely on the effect of EBP1 to reduce RBR1
protein level. We argue that EBP1 is a conserved, dose-dependent regulator of
cell growth that is connected to meristematic competence and cell proliferation
via regulation of RBR1 level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17024182 [PubMed - indexed for MEDLINE]

1060: Plant Physiol Biochem. 2006 Jul-Sep;44(7-9):483-93. Epub 2006 Sep 5.

Structural and functional analysis of a salt stress inducible gene encoding
voltage dependent anion channel (VDAC) from pearl millet (Pennisetum glaucum).

Desai MK, Mishra RN, Verma D, Nair S, Sopory SK, Reddy MK.

International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali
Marg, New Delhi 110 067, India.

We have cloned and characterized a gene encoding voltage-dependent anion channel
from Pennisetum glaucum (PgVDAC). PgVDAC was identified while isolating genes
that were differentially up-regulated following salt stress. The genomic
organization of PgVDAC clone was well conserved compared to other plant VDAC
genes in terms of number of introns, their position and phasing, however, the
primary amino acid sequence of voltage dependent anion channel (VDAC) proteins
did not show much conservation with other plant VDACs but their secondary and
tertiary structures are well conserved as predicted by in silico structural and
CD spectra analyses and results show it to be a typical membrane-spanning
beta-barrel leading to the formation of pore in the membrane. The heterologous
expression of PgVDAC protein in yeast strain lacking the endogenous mitochondrial
VDAC gene could not functionally complement it as was also previously observed
for the potato VDAC. Using real-time quantitative PCR analysis it was found that
transcript expression profile of PgVDAC was quantitatively and kinetically
up-regulated in response to salinity, desiccation, cold and exogenous application
of salicylic acid (SA); however, there was no effect of exogenous application of
abscisic acid (ABA) on its expression. Constitutive over-expression of PgVDAC
appears to be deleterious in transgenic rice plant; however, low level of
up-regulation imparted salinity stress adaptive response. A search for a more
suitable inducible transgene system is currently under way to understand PgVDAC
expression levels in plant development and its role in stress adaptation.

PMID: 17023166 [PubMed - indexed for MEDLINE]

1061: Prikl Biokhim Mikrobiol. 2006 Jul-Aug;42(4):485-8.

[Accuracy of a real-time polymerase-chain-reaction assay for a quantitative
estimation of genetically modified sources in food products]

[Article in Russian]

Abramov DD, Trofimov DIu, Rebrikov DV.

The accuracy of a real-time polymerase-chain-reaction assay for genetically
modified sources in food products was determined using two official test systems
(kits) of primers and samples. These kits were recommended by the Federal Center
of State Sanitary and Epidemiological Surveillance (Russian Ministry of Health)
and the European Commission. We used the following three models of thermocyclers:
iCycler iQ (BioRad, United States), Rotor-Gene 3000 (Corbett Research,
Australia), and DT-322 (DNA-Technology, Russia). Studies of samples that
contained 1% genetically modified sources showed that the error of a quantitative
assay for genetically modified sources in food products corresponds to 20-30% and
does not depend on the kit type and the thermocycler model used.

Publication Types:
Comparative Study
English Abstract

PMID: 17022461 [PubMed - indexed for MEDLINE]

1062: Prikl Biokhim Mikrobiol. 2006 Jul-Aug;42(4):462-7.

[The effect of thaumatin gene overexpression on the properties of H(+)-ATPase
from the plasmalemma of potato tuber cells]

[Article in Russian]

Ladyzhenskaia EP, Korableva NP.

The introduction of the thaumatin gene into potato plants was accompanied by a
decrease in the activity of H(+)-ATPase in the plasmalemma (PL) of tuber cells.
When tubers were released from dormancy, the enzyme was activated in the tuber
cells of both the original and transgenic plants. Experiments performed in vitro
demonstrated that sensitivities to ambiol (AM) and jasmonic acid (JA) of
H(+)-ATPase in the PL of tubers from the original plants were lower after the
release from a period of deep dormancy. In preparations from the tubers of
transgenic plants, the situation was reversed. The differences between the
activities of H(+)-ATPase in the PL preparations produced from the original and
transgenic tubers that sprouted under the action of AM and JA were detected.
Thus, the overexpression of the thaumatin gene in potato plants changed the
properties of H(+)-ATPase from PL.

Publication Types:
English Abstract

PMID: 17022457 [PubMed - indexed for MEDLINE]

1063: Environ Toxicol Chem. 2006 Oct;25(10):2653-61.

Subacute effects of transgenic crylab bacillus thuringiensis corn litter on the
isopods Trachelipus rathkii and armadillidium nasatum.

Clark BW, Prihoda KR, Coats JR.

Department of Entomology and Interdepartmental Toxicology Program, Iowa State
University, 115 Insectary, Ames, Iowa 50011, USA.

Laboratory studies were conducted to investigate the subacute effects of
transgenic Cry1Ab corn leaf material containing Bacillus thuringiensis (Bt)
protein on the terrestrial isopods Trachelipus rathkii and Armadillidium nasatum.
Survival and growth were measured for eight weeks in isopods fed leaf material of
two Bt11 corn varieties, two Monsanto 810 (Mon810) corn varieties, and the
isolines of each. Total lipid and protein content of the organisms was measured
to examine effects on energetic reserves. Armadillidium nasatum individuals in
all treatments responded similarly. For T. rathkii, no statistically significant
effect of Bt was observed, but statistical differences were observed in growth
between hybrids. Protein and sugar content of the food were found to be
correlated with the differences in growth for T. rathkii. Total protein content
was higher in T. rathkii and A. nasatum fed material with higher protein and
sugar content. A trend toward less growth in T. rathkii on Bt corn varieties
versus their isolines triggered a concentration-response assay with purified
Cry1Ab protein. No adverse effects of purified Bt protein were observed. These
results indicate that little hazard to T. rathkii and A. nasatum from Bt corn
leaf material from these hybrids exists. However, nutritional differences in corn
hybrids contributed to differences in isopod growth.

PMID: 17022406 [PubMed - indexed for MEDLINE]

1064: Mol Plant Microbe Interact. 2006 Oct;19(10):1127-37.

Inducible overexpression of a rice allene oxide synthase gene increases the
endogenous jasmonic acid level, PR gene expression, and host resistance to fungal
infection.

Mei C, Qi M, Sheng G, Yang Y.

Department of Plant Pathology and Program in Cell and Molecular Biology,
University of Arkansas, Fayetteville 72071, USA.

Many studies in dicotyledonous plants have shown that jasmonates, including
jasmonic acid (JA) and methyl jasmonate, are important signal molecules involved
in induced resistance to pathogen infection and insect herbivory. However, very
little genetic and molecular evidence is available to demonstrate their role in
host defense response of rice and other economically important monocot plants. In
this study, we have shown that exogenous application of JA was able to activate
defense gene expression and local induced resistance in rice seedlings against
the rice blast fungus (Magnaporthe grisea). Furthermore, we have characterized a
pathogen-inducible rice OsAOS2 gene (which encodes allene oxide synthase, a key
enzyme in the JA biosynthetic pathway) and examined the role of endogenous JA in
rice defense response through transgenic manipulation of the JA biosynthesis.
Sequence analysis indicated that OsAOS2 contains four common domains of the
cytochrome P450 enzyme, but does not have the signal peptide for chloroplast
targeting. The basal level of OsAOS2 expression is very low in leaves but
relatively high in the sheath, culm, and flower of rice plants. Interestingly,
the expression of OsAOS2 in rice leaves can be induced significantly upon M.
grisea infection. Transgenic rice lines carrying the OsAOS2 transgene under the
control of a strong, pathogen-inducible PBZ1 promoter accumulated abundant OsAOS2
transcripts and higher levels of JA, especially after the pathogen infection.
These transgenic lines also exhibited enhanced activation of pathogenesis-related
(PR) genes such as PR1a, PR3, and PR5 and increased resistance to M. grisea
infection. Our results suggest that JA plays a significant role in PR gene
induction and blast resistance in rice plants.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17022177 [PubMed - indexed for MEDLINE]

1065: Plant Mol Biol. 2007 Jan;63(1):73-84. Epub 2006 Oct 5.

Wheat Dof transcription factor WPBF interacts with TaQM and activates
transcription of an alpha-gliadin gene during wheat seed development.

Dong G, Ni Z, Yao Y, Nie X, Sun Q.

Department of Plant Genetics & Breeding and State Key Laboratory for
Agrobiotechnology, China Agricultural University, Haidian district, Beijing,
100094, China.

Wheat prolamin-box binding factor (WPBF), a DOF transcription factor previously
was isolated from wheat endosperm and suggested to function as an activator of
prolamin gene expression during seed development. In this study, we showed that
WPBF is expressed in all wheat tissues analyzed, and a protein, TaQM, was
identified from a wheat root cDNA library, to interact with the Dof domain of
WPBF. The specific interaction between WPBF and TaQM was confirmed by pull-down
assay and bimolecular fluorescence complementation (BiFC) experiment. The
expression patterns of TaQM gene are similar with that of WPBF. The GST-WPBF
expressed in bacteria binds the Prolamin box (PB) 5'-TGTAAAG-3', derived from the
promoter region of a native alpha-gliadin gene encoding a storage protein.
Transient expression experiments in co-transfected BY-2 protoplast cells
demonstrated that WPBF trans-activated transcription from native alpha-gliadin
promoter through binding to the intact PB. When WPBF and TaQM are co-transfected
together the transcription activity of alpha-gliadin gene was six-fold higher
than when WPBF was transfected alone. Furthermore, the promoter activities of
WPBF gene were observed in the seeds and the vascular system of transgenic
Arabidopsis, which was identical to the expression profiles of WPBF in wheat.
Hence, we proposed that WPBF functions not only during wheat seed development but
also during other growth and development processes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17021941 [PubMed - indexed for MEDLINE]

1066: Appl Microbiol Biotechnol. 2007 Jan;73(6):1470-6. Epub 2006 Oct 5.

Cryopreservation of transgenic rice suspension cells producing recombinant
hCTLA4Ig.

Cho JS, Hong SM, Joo SY, Yoo JS, Kim DI.

Department of Biological Engineering, Inha University, Incheon, 402-751, Republic
of Korea.

Transgenic suspension cells of Oryza sativa L. cv. Dongjin utilized as a host for
producing recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin
(hCTLA4Ig) were preserved in liquid nitrogen (-196 degrees C) after slow
prefreezing in a deep freezer (-70 degrees C). The development of an optimal
procedure for long-term storage was investigated by the addition of various
concentrations of cryoprotectant mixture and osmoticum in preculture media before
cooling. A pre-deep-freezing time of 120 min was the most effective for
maintaining cell viability. Compared with mannitol, sorbitol, trehalose, and NaCl
under the same osmotic conditions, 0.5 M sucrose was found to be the best
osmoticum for preculture media. The cryoprotectant comprising sucrose, glycerol,
and dimethylsulfoxide (DMSO) was applied to the precultured cells, and a
combination of 1 M sucrose, 1 M glycerol, and 1 M DMSO provided the best result.
The viability with this optimized condition was 88% after cryocell-banking for 1
day. The expression of hCTLA4Ig in recovered callus from cryopreservation was
also kept stable, and the production level was similar to that observed in
noncryopreserved cultures.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17021872 [PubMed - indexed for MEDLINE]

1067: Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18828-33. Epub 2006 Oct 4.

Artificial evolution extends the spectrum of viruses that are targeted by a
disease-resistance gene from potato.

Farnham G, Baulcombe DC.

The Sainsbury Laboratory, John Innes Centre, Norwich NR4 7UH, United Kingdom.

A major class of disease-resistance (R) genes in plants encode nucleotide-binding
site/leucine-rich repeat (LRR) proteins. The LRR domains mediate recognition of
pathogen-derived elicitors. Here we describe a random in vitro mutation analysis
illustrating how mutations in an R protein (Rx) LRR domain generate
disease-resistance specificity. The original Rx protein confers resistance only
against a subset of potato virus X (PVX) strains, whereas selected mutants were
effective against an additional strain of PVX and against the distantly related
poplar mosaic virus. These effects of LRR mutations indicate that in vitro
evolution of R genes could be exploited for enhancement of disease resistance in
crop plants. Our results also illustrate how short-term evolution of disease
resistance in wild populations might be toward broader spectrum resistance
against multiple strains of the pathogen. The breadth of the disease-resistance
phenotype from a natural R gene may be influenced by the tradeoff between the
costs and benefits of broad-spectrum disease resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17021014 [PubMed - indexed for MEDLINE]

1068: J Vet Med Sci. 2006 Sep;68(9):959-65.

Involvement of neuropeptide Y in hyperphagia in human growth hormone transgenic
rats.

Hozumi H, Yamanouchi K, Nishihara M.

Department of Veterinary Physiology, Veterinary Medical Science, The University
of Tokyo, Tokyo, Japan.

We have previously produced human growth hormone (hGH) transgenic (TG) rats that
show low circulating levels of both hGH and endogenous rat GH. Although body
length of the TG rats is normal, they develop hyperphagia and severe obesity. The
present study was undertaken to elucidate the causes of hyperphagia in the TG
rats by focusing on temporal changes in plasma ghrelin levels and hypothalamic
neuropeptide Y (NPY) contents. In both wild-type (WT) and TG rats, the highest
value of plasma ghrelin levels was observed just before the dark phase, and
thereafter plasma ghrelin levels were maintained higher in the TG than WT rats.
Although NPY contents also showed the peak level just before the dark phase in
both the arcuate (ARC) and paraventricular nuclei (PVN) of the hypothalamus, the
values in the ARC, but not the PVN, of the TG rats was always lower than those of
the WT rats, suggesting increased transport of NPY from the ARC to PVN in the TG
rats. In addition, treatment with antagonists for Y1 and Y5 receptors for NPY
reduced food intake much more effectively in the TG than WT rats. Intermittent
treatment with recombinant hGH for a week significantly decreased food
consumption, adipose tissue weight and plasma triglyceride concentrations in the
TG rats. These results suggest that, in the TG rats, insufficiency in circulating
GH stimulates the ghrelin-NPY system with a resultant increase in food intake.

PMID: 17019066 [PubMed - indexed for MEDLINE]

1069: Plant Cell Rep. 2007 Mar;26(3):313-25. Epub 2006 Oct 3.

The PsEND1 promoter: a novel tool to produce genetically engineered male-sterile
plants by early anther ablation.

Roque E, Gö¨mez MD, Ellul P, Wallbraun M, Madueö±o F, Beltrö¡n JP, Caö±as LA.

Departamento de Biologö­a del Desarrollo, Instituto de Biologö­a Molecular y
Celular de Plantas (C.S.I.C.-U.P.V.), Campus de la Universidad Politö©cnica de
Valencia, Avda. de los Naranjos s/n., 46022 Valencia, Spain.

PsEND1 is a pea anther-specific gene that displays very early expression in the
anther primordium cells. Later on, PsEND1 expression becomes restricted to the
epidermis, connective, endothecium and middle layer, but it is never observed in
tapetal cells or microsporocytes. We fused the PsEND1 promoter region to the
cytotoxic barnase gene to induce specific ablation of the cell layers where the
PsEND1 is expressed and consequently to produce male-sterile plants. Expression
of the chimaeric PsEND1::barnase gene in two Solanaceae (Nicotiana tabacum and
Solanum lycopersicon) and two Brassicaceae (Arabidopsis thaliana and Brassica
napus) species, impairs anther development from very early stages and produces
complete male-sterile plants. The PsEND1::barnase gene is quite different to
other chimaeric genes previously used in similar approaches to obtain
male-sterile plants. The novelty resides in the use of the PsEND1 promoter,
instead of a tapetum-specific promoter, to produce the ablation of specific cell
lines during the first steps of the anther development. This chimaeric construct
arrests the microsporogenesis before differentiation of the microspore mother
cells and no viable pollen grains are produced. This strategy represents an
excellent alternative to generate genetically engineered male-sterile plants,
which have proved useful in breeding programmes for the production of hybrid
seeds. The PsEND1 promoter also has high potential to prevent undesirable
horizontal gene flow in many plant species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17016735 [PubMed - indexed for MEDLINE]

1070: Plant Cell Rep. 2007 Feb;26(2):247-52. Epub 2006 Oct 3.

Transgenic Indian mustard (Brassica juncea) expressing tomato glucanase leads to
arrested growth of Alternaria brassicae.

Mondal KK, Bhattacharya RC, Koundal KR, Chatterjee SC.

Department of Plant Pathology, Chaudhary Swaran Kumar Himachal Pradesh
Agricultural University, Palampur, 176062, Himachal Pradesh, India.
kalyanmondal@yahoo.com

Brassica juncea is an important oilseed crop of the Indian sub-continent. Yield
loss due to fungal disease alternaria leaf spot caused by Alternaria brassicae is
a serious problem in cultivation of this crop. Nonavailability of resistance
genes within crossable germplasms of Brassica necessitates use of genetic
engineering strategies to develop genetic resistance against this pathogen. The
pathogenesis related (PR) proteins are group of plant proteins that are toxic to
invading fungal pathogens, but are present in plant in trace amount. Thus,
overexpression of PR proteins leads to increased resistance to pathogenic fungi
in several crops. The PR protein glucanase hydrolyzes a major cell-wall
component, glucan, of pathogenic fungi and acts as a plant defense barrier. We
report the expression of a class I basic glucanase gene, under the control of
CaMV 35S promoter, in Indian mustard and its genetic resistance against
alternaria leaf spot. Southern and Northern hybridization confirmed stable
integration and expression of the glucanase gene in mustard transgenics. Several
independent transgenics were screened in vitro and under poly house conditions
for their resistance against Alternaria brassicae. In an in vitro antifungal
assay, transgenics arrested hyphal growth of Alternaria brassicae by 15-54%.
Under pathogen-challenged conditions in poly house, the transgenics showed
restricted number, size and spread of lesions caused by Alternaria brassicae.
Also, the onset of disease was delayed in transgenics compared to untransformed
parent plants. The results demonstrate potentiality of a PR protein from a
heterologous source in developing alternaria leaf spot resistance in Indian
mustard.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17016733 [PubMed - indexed for MEDLINE]

1071: Proc Biol Sci. 2006 Dec 22;273(1605):3111-5.

Spontaneous gene flow from rapeseed (Brassica napus) to wild Brassica oleracea.

Ford CS, Allainguillaume J, Grilli-Chantler P, Cuccato G, Allender CJ, Wilkinson
MJ.

School of Biological Sciences, Lyle Building, The University of Reading,
Whiteknights, Reading RG6 6AS, UK.

Research on the environmental risks of gene flow from genetically modified (GM)
crops to wild relatives has traditionally emphasized recipients yielding most
hybrids. For GM rapeseed (Brassica napus), interest has centred on the
'frequently hybridizing' Brassica rapa over relatives such as Brassica oleracea,
where spontaneous hybrids are unreported in the wild. In two sites, where
rapeseed and wild B. oleracea grow together, we used flow cytometry and
crop-specific microsatellite markers to identify one triploid F1 hybrid, together
with nine diploid and two near triploid introgressants. Given the newly
discovered capacity for spontaneous introgression into B. oleracea, we then
surveyed associated flora and fauna to evaluate the capacity of both recipients
to harm cohabitant species with acknowledged conservational importance. Only B.
oleracea occupies rich communities containing species afforded legislative
protection; these include one rare micromoth species that feeds on B. oleracea
and warrants further assessment. We conclude that increased attention should now
focus on B. oleracea and similar species that yield few crop-hybrids, but possess
scope to affect rare or endangered associates.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17015343 [PubMed - indexed for MEDLINE]

1072: Med Hypotheses. 2007;68(1):22-30. Epub 2006 Oct 2.

Comment in:
Med Hypotheses. 2007;68(5):1180-1.

An edible vaccine for malaria using transgenic tomatoes of varying sizes, shapes
and colors to carry different antigens.

Chowdhury K, Bagasra O.

Department of Biology, South Carolina Center for Biotechnology, Claflin
University, 400 Magnolia Street, Orangeburg, SC 29115, USA.

Malaria, a disease caused by protozoan parasites of genus Plasmodium, is one of
the world's biggest scourges. Over two billion individuals reside in the malaria
endemic areas and the disease affects 300-500 million people annually. As a
result of malarial-infection, an estimated three million lives are lost annually,
among them over one million children (majority under 5 years of age). The
mortality due to malaria has increased because of the spread of drug-resistant
strains of the parasite, the breakdown of health services in many affected areas,
the interaction of the disease with human immunodeficiency virus (HIV) infection,
and possibly the effects of climate change. Infants and young children with
malaria often die from severe anemia, cerebral involvement,or prostration caused
by overwhelming infection; many new borns die from complications of low birth
weight caused by maternal malaria during pregnancy.The scarce economic resources
and lack of communication, infrastructure and adequate means of travel in the
endemic areas make it extremely difficult to implement traditional infection
control measures (i.e., mosquito control, preventive anti-malarial drugs and
nets). To make the matter worse, both malarial parasites and its insect vectors
are increasingly becoming resistant to anti-malarial agents (chloroquine) and
insecticides (both DDT and melathione and related chemicals), respectively. By
conventional wisdom, the immune mechanisms responsible for protection against
malaria will require a multiple of 10-15 antigen targets for proper protection
against various stages of malarial infection. By standard vaccination protocols,
such a large number of targets would not be appropriate to be used for
vaccination as a single dose due to antigenic competition. It would be almost
impossible to immunize over two billion individuals who live in malaria
susceptible areas with several carefully crafted immunization schedules delivered
4-6 weeks apart in the form of two different antigens as a single dose. Besides,
if immunization schedules could be arranged, the stability of vaccines carrying
different malarial antigens, their transport, and the logistics of vaccination
would be an almost impossible task to achieve under the current fiscal
constraints. We are proposing a unique way to circumvent these logistical
difficulties to deliver the malaria vaccines to every susceptible home at a small
fraction of a cost. We hypothesize that the anti-malaria edible vaccines in
transgenic tomato plants where different transgenic plants expressing different
antigenic type(s). Immunizing individuals against 2-3 antigens and against each
stage of the life cycle of the multistage parasites would be an efficient,
inexpensive and safe way of vaccination. Tomatoes with varying sizes, shapes and
colors carrying different antigens would make the vaccines easily identifiable by
lay individuals.

PMID: 17014967 [PubMed - indexed for MEDLINE]

1073: Plant Physiol. 2006 Nov;142(3):1053-64. Epub 2006 Sep 29.

The cyclin-dependent kinase inhibitor Orysa;KRP1 plays an important role in seed
development of rice.

Barrö´co RM, Peres A, Droual AM, De Veylder L, Nguyen le SL, De Wolf J, Mironov V,
Peerbolte R, Beemster GT, Inzö© D, Broekaert WF, Frankard V.

Department of Plant Systems Biology, Flanders Interuniversity Institute for
Biotechnology, Ghent University, B-9052 Ghent, Belgium.

Kip-related proteins (KRPs) play a major role in the regulation of the plant cell
cycle. We report the identification of five putative rice (Oryza sativa) proteins
that share characteristic motifs with previously described plant KRPs. To
investigate the function of KRPs in rice development, we generated transgenic
plants overexpressing the Orysa;KRP1 gene. Phenotypic analysis revealed that
overexpressed KRP1 reduced cell production during leaf development. The reduced
cell production in the leaf meristem was partly compensated by an increased cell
size, demonstrating the existence of a compensatory mechanism in monocot species
by which growth rate is less reduced than cell production, through cell
expansion. Furthermore, Orysa;KRP1 overexpression dramatically reduced seed
filling. Sectioning through the overexpressed KRP1 seeds showed that KRP
overproduction disturbed the production of endosperm cells. The decrease in the
number of fully formed seeds was accompanied by a drop in the endoreduplication
of endosperm cells, pointing toward a role of KRP1 in connecting endocycle with
endosperm development. Also, spatial and temporal transcript detection in
developing seeds suggests that Orysa;KRP1 plays an important role in the exit
from the mitotic cell cycle during rice grain formation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17012406 [PubMed - indexed for MEDLINE]

1074: Plant Physiol. 2006 Nov;142(3):1160-8. Epub 2006 Sep 29.

Regulation of seed size by hypomethylation of maternal and paternal genomes.

Xiao W, Brown RC, Lemmon BE, Harada JJ, Goldberg RB, Fischer RL.

Department of Plant and Microbial Biology, University of California, Berkeley,
California 94720, USA.

DNA methylation is an epigenetic modification of cytosine that is important for
silencing gene transcription and transposons, gene imprinting, development, and
seed viability. DNA METHYLTRANSFERASE1 (MET1) is the primary maintenance DNA
methyltransferase in Arabidopsis (Arabidopsis thaliana). Reciprocal crosses
between antisense MET1 transgenic and wild-type plants show that DNA
hypomethylation has a parent-of-origin effect on seed size. However, due to the
dominant nature of the antisense MET1 transgene, the parent with a hypomethylated
genome, its gametophyte, and both the maternal and paternal genomes of the F(1)
seed become hypomethylated. Thus, the distinct role played by hypomethylation at
each generation is not known. To address this issue, we examined F(1) seed from
reciprocal crosses using a loss-of-function recessive null allele, met1-6.
Crosses between wild-type and homozygous met1-6 parents show that hypomethylated
maternal and paternal genomes result in significantly larger and smaller F(1)
seeds, respectively. Our analysis of crosses between wild-type and heterozygous
MET1/met1-6 parents revealed that hypomethylation in the female or male
gametophytic generation was sufficient to influence F(1) seed size. A recessive
mutation in another gene that dramatically reduces DNA methylation, DECREASE IN
DNA METHYLATION1, also causes parent-of-origin effects on F(1) seed size. By
contrast, recessive mutations in genes that regulate a smaller subset of DNA
methylation (CHROMOMETHYLASE3 and DOMAINS REARRANGED METHYLTRANSFERASES1 and 2)
had little effect on seed size. Collectively, these results show that maternal
and paternal genomes play distinct roles in the regulation of seed size in
Arabidopsis.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 17012404 [PubMed - indexed for MEDLINE]

1075: PLoS Comput Biol. 2006 Sep 29;2(9):e128. Epub 2006 Aug 21.

Can transgenic maize affect soil microbial communities?

Mulder C, Wouterse M, Raubuch M, Roelofs W, Rutgers M.

Laboratory for Ecological Risk Assessment, National Institute for Public Health
and the Environment, Bilthoven, Netherlands. christian.mulder@rivm.nl

The aim of the experiment was to determine if temporal variations of belowground
activity reflect the influence of the Cry1Ab protein from transgenic maize on
soil bacteria and, hence, on a regulatory change of the microbial community
(ability to metabolize sources belonging to different chemical guilds) and/or a
change in numerical abundance of their cells. Litter placement is known for its
strong influence on the soil decomposer communities. The effects of the addition
of crop residues on respiration and catabolic activities of the bacterial
community were examined in microcosm experiments. Four cultivars of Zea mays L.
of two different isolines (each one including the conventional crop and its
Bacillus thuringiensis cultivar) and one control of bulk soil were included in
the experimental design. The growth models suggest a dichotomy between soils
amended with either conventional or transgenic maize residues. The Cry1Ab protein
appeared to influence the composition of the microbial community. The highly
enhanced soil respiration observed during the first 72 h after the addition of
Bt-maize residues can be interpreted as being related to the presence of the
transgenic crop residues. This result was confirmed by agar plate counting, as
the averages of the colony-forming units of soils in conventional treatments were
about one-third of those treated with transgenic straw. Furthermore, the addition
of Bt-maize appeared to induce increased microbial consumption of carbohydrates
in BIOLOG EcoPlates. Three weeks after the addition of maize residues to the
soils, no differences between the consumption rate of specific chemical guilds by
bacteria in soils amended with transgenic maize and bacteria in soils amended
with conventional maize were detectable. Reaped crop residues, comparable to
post-harvest maize straw (a common practice in current agriculture), rapidly
influence the soil bacterial cells at a functional level. Overall, these data
support the existence of short Bt-induced ecological shifts in the microbial
communities of croplands' soils.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17009863 [PubMed - indexed for MEDLINE]

1076: Plant Mol Biol. 2006 Dec;62(6):897-912. Epub 2006 Sep 28.

Characterization of the plant Notchless homolog, a WD repeat protein involved in
seed development.

Chantha SC, Emerald BS, Matton DP.

Institut de Recherche en Biologie Vö©gö©tale (IRBV), Dö©partement de sciences
biologiques, Universitö© de Montrö©al, 4101 rue Sherbrooke est, H1X 2B2, Montrö©al,
QC, Canada.

We have isolated a plant NOTCHLESS (NLE) homolog from the wild potato species
Solanum chacoense Bitt., encoding a WD-repeat containing protein initially
characterized as a negative regulator of the Notch receptor in animals. Although
no Notch signaling pathway exists in plants, the NLE gene is conserved in
animals, plants, and yeast. Overexpression of the plant ScNLE gene in Drosophila
similarly affected bristle formation when compared to the overexpression of the
endogenous Drosophila NLE gene, suggesting functional conservation. Expression
analyses showed that the ScNLE gene was fertilization-induced and primarily
expressed in ovules after fertilization, mainly in the integumentary tapetum
(endothelium). Significant expression was also detected in the shoot apex.
Promoter deletion analysis revealed that the ScNLE promoter had a complex
modulatory architecture with both positive, negative, and tissue specific
regulatory elements. Transgenic plants with reduced levels of ScNLE transcripts
displayed pleitotropic phenotypes including a severe reduction in seed set,
consistent with ScNLE gene expression pattern.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17006595 [PubMed - indexed for MEDLINE]

1077: Plant Mol Biol. 2007 Jan;63(1):21-34. Epub 2006 Sep 28.

Comparative expression profiling in meristems of inbred-hybrid triplets of maize
based on morphological investigations of heterosis for plant height.

Uzarowska A, Keller B, Piepho HP, Schwarz G, Ingvardsen C, Wenzel G, Lö¼bberstedt
T.

Department of Plant Breeding, Technical University of Munich, Freising, Germany.

Heterosis, the superior performance of hybrids as compared to their parental mean
is an agronomically important phenomenon well-described morphologically. However,
little is known about its molecular basis. We investigated four genetically
unrelated maize (Zea mays L.) inbred lines and their F(1) crosses both at the
phenotype and transcriptome level, focusing on plant height (PHT) component
traits. Substantial mid-parent heterosis (MPH) was found for all parent-hybrid
triplets for PHT in the range of 37.9-56.4% in the field and 11.1-39.5% under
controlled greenhouse conditions. Analyses of heterosis for number and length of
internodes showed two to three times higher MPH in the field as compared to the
greenhouse. All three traits exhibited high heritabilities, highest for PHT
95-98%. Two methods for gene expression quantification were applied. High-density
cDNA uni-gene microarrays containing 11,827 ESTs were utilized for the selection
of differentially expressed genes related to heterosis for PHT. For the four
triplets with eight possible parent-hybrid comparisons we identified 434
consistently differentially expressed genes with a p < or = 0.05. Microarray
results were used to verify the dominance/overdominance hypothesis. In our study,
more than 50% genes showed overdominance, 26% partial dominance, 12.6% complete
dominance and 10.2% additive gene action. Moreover, more consistently
differentially expressed genes were detected in related triplets, sharing one
parent, than in unrelated triplets. Quantitative RT-PCR was applied in order to
validate microarray results. The role of the differentially expressed genes in
relation to heterosis for PHT is discussed.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17006594 [PubMed - indexed for MEDLINE]

1078: Heredity. 2006 Dec;97(6):379-80. Epub 2006 Sep 27.

Polluting gene flow from crops: radishes gone wild.

Chapman MA, Burke JM.

Publication Types:
News

PMID: 17006535 [PubMed - indexed for MEDLINE]

1079: J Biosci. 2006 Sep;31(3):339-45.

Agrobacterium-mediated transformation of chickpea with alpha-amylase inhibitor
gene for insect resistance.

Ignacimuthu S, Prakash S.

Entomology Research Institute, Loyola College, Chennai 600 034, India.
eri_lc@hotmail.com

Chickpea is the world's third most important pulse crop and India produces 75% of
the world's supply. Chickpea seeds are attacked by Callosobruchus maculatus and
C. chinensis which cause extensive damage. The alpha-amylase inhibitor gene
isolated from Phaseolus vulgaris seeds was introduced into chickpea cultivar K850
through Agrobacterium-mediated transformation. A total of 288 kanamycin resistant
plants were regenerated. Only 0.3% of these were true transformants. Polymerase
chain reaction (PCR) analysis and Southern hybridization confirmed the presence
of 4.9 kb alpha-amylase inhibitor gene in the transformed plants. Western blot
confirmed the presence of alpha-amylase inhibitor protein. The results of
bioassay study revealed a significant reduction in the survival rate of bruchid
weevil C. maculatus reared on transgenic chickpea seeds. All the transgenic
plants exhibited a segregation ratio of 3:1.

PMID: 17006016 [PubMed - indexed for MEDLINE]

1080: Acta Trop. 2006 Oct;99(2-3):173-83. Epub 2006 Sep 26.

Re-introducing bacteria in mosquitoes--a method for determination of mosquito
feeding preferences based on coloured sugar solutions.

Lindh JM, Terenius O, Eriksson-Gonzales K, Knols BG, Faye I.

Department of Genetics, Microbiology and Toxicology, Stockholm University, 106 91
Stockholm, Sweden.

In this study, sugar-feeding was investigated as a possible means of
re-introducing bacteria into mosquito midguts with the aim of identifying
bacteria that are suitable for creating paratransgenic mosquitoes. In a
paratransgenic approach, bacteria are utilised to deliver effector molecules
capable of inhibiting pathogen development in the midgut of the vector. To
determine if mosquitoes discriminate between sterile sugar solutions and sugar
solutions with bacteria, a method for screening mosquito feeding preferences was
developed. This method was tested for Aedes aegypti, Anopheles arabiensis and An.
gambiae s.s. mosquitoes and is based on a dual-choice test of solutions labelled
with food dyes. Three different tests (dye/colour detection, sugar detection and
sugar-concentration detection) were performed to evaluate the method, after which
bacteria previously isolated from mosquitoes were used in the experiments. It was
shown that mosquitoes do not discriminate between sugar solutions with or without
these bacteria indicating that sugar-feeding is a possible means to introduce
bacteria into mosquitoes. Furthermore, two different setups of the method were
used, enabling us to differentiate between tactile/taste and olfactory responses.
The method described in this paper is easy to use, cost-effective and allows
broad screening of mosquito sugar-feeding preferences.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16999928 [PubMed - indexed for MEDLINE]

1081: Nat Cell Biol. 2006 Oct;8(10):1143-8. Epub 2006 Sep 24.

Oocyte signals derived from polyunsaturated fatty acids control sperm recruitment
in vivo.

Kubagawa HM, Watts JL, Corrigan C, Edmonds JW, Sztul E, Browse J, Miller MA.

Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL
35294, USA.

A fundamental question in animal development is how motile cells find their
correct target destinations. During mating in the nematode Caenorhabditis
elegans, males inject sperm through the hermaphrodite vulva into the uterus.
Amoeboid sperm crawl around fertilized eggs to the spermatheca--a convoluted tube
where fertilization occurs. Here, we show that polyunsaturated fatty acids
(PUFAs), the precursors of eicosanoid signalling molecules, function in oocytes
to control directional sperm motility within the uterus. PUFAs are transported
from the intestine, the site of fat metabolism, to the oocytes yolk, which is a
lipoprotein complex. Loss of the RME-2 low-density lipoprotein (LDL) receptor,
which mediates yolk endocytosis and fatty acid transport into oocytes, causes
severe defects in sperm targeting. We used an RNAi screen to identify lipid
regulators required for directional sperm motility. Our results support the
hypothesis that PUFAs function in oocytes as precursors of signals that control
sperm recruitment to the spermatheca. A common property of PUFAs in mammals and
C. elegans is that these fats control local recruitment of motile cells to their
target tissues.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 16998478 [PubMed - indexed for MEDLINE]

1082: Plant Physiol. 2006 Nov;142(3):1102-12. Epub 2006 Sep 22.

Improved resistance to controlled deterioration in transgenic seeds.

Prieto-Dapena P, Castaö±o R, Almoguera C, Jordano J.

Instituto de Recursos Naturales y Agrobiologö­a, Consejo Superior de
Investigaciones Cientö­ficas, 41080 Seville, Spain.

We show that seed-specific overexpression of the sunflower (Helianthus annuus)
HaHSFA9 heat stress transcription factor (HSF) in tobacco (Nicotiana tabacum)
enhances the accumulation of heat shock proteins (HSPs). Among these proteins
were HSP101 and a subset of the small HSPs, including proteins that accumulate
only during embryogenesis in the absence of thermal stress. Levels of late
embryogenesis abundant proteins or seed oligosaccharides, however, were not
affected. In the transgenic seeds, a high basal thermotolerance persisted during
the early hours of imbibition. Transgenic seeds also showed significantly
improved resistance to controlled deterioration in a stable and
transgene-dependent manner. Furthermore, overexpression of HaHSFA9 did not have
detrimental effects on plant growth or development, including seed morphology and
total seed yield. Our results agree with previous work tentatively associating
HSP gene expression with phenotypes important for seed longevity. These findings
might have implications for improving seed longevity in economically important
crops.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16998084 [PubMed - indexed for MEDLINE]

1083: J Exp Bot. 2006;57(14):3933-43. Epub 2006 Sep 22.

Effect of ascorbate oxidase over-expression on ascorbate recycling gene
expression in response to agents imposing oxidative stress.

Fotopoulos V, Sanmartin M, Kanellis AK.

Group of Biotechnology of Pharmaceutical Plants, Laboratory of Pharmacognosy,
Department of Pharmaceutical Sciences, Aristotle University of Thessaloniki, 541
24 Thessaloniki, Greece.

Ascorbate oxidase (AO) is a cell wall-localized enzyme that uses oxygen to
catalyse the oxidation of ascorbate (AA) to the unstable radical
monodehydroascorbate (MDHA) which rapidly disproportionates to yield
dehydroascorbate (DHA) and AA, and thus contributes to the regulation of the AA
redox state. Here, it is reported that in vivo lowering of the apoplast AA redox
state, through increased AO expression in transgenic tobacco (Nicotiana tabacum
L. cv. Xanthi), exerts no effects on the expression levels of genes involved in
AA recycling under normal growth conditions, but plants display enhanced
sensitivity to various oxidative stress-promoting agents. RNA blot analyses
suggest that this response correlates with a general suppression of the plant's
antioxidative metabolism as demonstrated by lower expression levels of AA
recycling genes. Furthermore, studies using Botrytis cinerea reveal that
transgenic plants exhibit increased sensitivity to fungal infection, although the
response is not accompanied by a similar suppression of AA recycling gene
expression. Our current findings, combined with previous studies which showed the
contribution of AO in the regulation of AA redox state, suggest that the
reduction in the AA redox state in the leaf apoplast of these transgenic plants
results in shifts in their capacity to withstand oxidative stress imposed by
agents imposing oxidative stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16997902 [PubMed - indexed for MEDLINE]

1084: Science. 2006 Sep 22;313(5794):1714.

Genetically modified crops. Tracing the transatlantic spread of GM rice.

Vogel G.

Publication Types:
News

PMID: 16990520 [PubMed - indexed for MEDLINE]

1085: J Exp Bot. 2006;57(14):3717-26. Epub 2006 Sep 21.

Tissue-specific expression of tomato Ribonuclease LX during phosphate
starvation-induced root growth.

Kö¶ck M, Stenzel I, Zimmer A.

Martin Luther University Halle-Wittenberg, Biocenter, D-06120 Halle, Germany.
margret.koeck@biozentrum.uni-halle.de

Ribonuclease LX (RNaseLX) from tomato (Solanum lycopersicum L.) belongs to the
RNase T2/S-RNase superfamily of plant endoribonucleases and this is a report on
the characterization of the RNaseLX gene and its encoded protein as a member of
the phosphate starvation response in tomato. RNaseLX gene sequences were cloned
by a PCR-assisted approach. RNaseLX promoter sequences contained the conserved
binding motif of the transcription factor PHR1 known to mediate phosphate
starvation-dependent gene expression. The increase of RNaseLX transcript levels
in roots during phosphate starvation correlated with high promoter activity in
transgenic plants carrying a PromLX::uidA gene construct and pointed to
transcriptional control of RNaseLX expression. Histochemical staining for
beta-glucuronidase activity and immunodetection of RNaseLX protein revealed
striking RNaseLX expression in main and lateral root tips of phosphate-starved
transgenic plants, specifically in epidermal cells, as well as in lateral and
adventitious root primordia. Induced RNaseLX expression in roots correlated with
stimulated growth and elongation of primary and lateral roots during phosphate
deprivation. Phosphate-starvation-induced RNaseLX transcript levels in roots were
not modulated by auxin or ethylene. These data indicate that the role of
intracellular RNaseLX in the phosphate starvation response is connected with
specific RNA turnover processes at the root tip.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16990375 [PubMed - indexed for MEDLINE]

1086: Plant Cell Physiol. 2006 Nov;47(11):1457-72. Epub 2006 Sep 20.

Erratum in:
Plant Cell Physiol. 2007 Jan;48(1):204.

Rice Immature Pollen 1 (RIP1) is a regulator of late pollen development.

Han MJ, Jung KH, Yi G, Lee DY, An G.

National Research Laboratory of Plant Functional Genomics, Division of Molecular
and Life Sciences, Republic of Korea.

We isolated a pollen-preferential gene, RICE IMMATURE POLLEN 1 (RIP1), from a
T-DNA insertional population of japonica rice that was trapped by a promoterless
beta-glucuronidase (GUS) gene. Semi-quantitative reverse transcription-PCR
(RT-PCR) analyses confirmed that the RIP1 transcript was abundant at the late
stages of pollen development. Transgenic plants carrying a T-DNA insertion in the
RIP1 gene displayed the phenotype of segregation distortion of the mutated rip1
gene. Moreover, rip1/rip1 homozygous progeny were not present. Reciprocal crosses
between Rip1/rip1 heterozygous plants and the wild type showed that the rip1
allele could not be transmitted through the male. Microscopic analysis
demonstrated that development in the rip1 pollen was delayed, starting at the
early vacuolated stage. Close examination of that pollen by transmission electron
microscopy also showed delayed formation of starch granules and the intine layer.
In addition, development of the mitochondria, Golgi apparatus, lipid bodies,
plastids and endoplasmic reticulum was deferred in the mutant pollen. Under in
vitro conditions, germination of this mutant pollen did not occur, whereas the
rate for wild-type pollen was >90%. These results indicate that RIP1 is necessary
for pollen maturation and germination. This gene encodes a protein that shares
significant homology with a group of proteins containing five WD40 repeat
sequences. The green fluorescent protein (GFP)-RIP1 fusion protein is localized
to the nucleus. Therefore, RIP1 is probably a nuclear protein that may form a
functional complex with other proteins and carry out essential cellular and
developmental roles during the late stage of pollen formation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16990291 [PubMed - indexed for MEDLINE]

1087: Sci China C Life Sci. 2006 Aug;49(4):305-10.

Hairy roots induced by Agrobacterium rhizogenes and production of regenerative
plants in hairy root cultures in maize.

Xu H, Zhou X, Lu J, Wang J, Wang X.

School of Life Sciences, Northeast Normal University, Changchun 130024, China.

Hairy roots of maize were induced by infecting 15-d calli with Agrobacterium
rhizogenes. The hairy roots cultured in hormone-free media showed the vigorous
growth and typical hairy root features. The regenerated plants were produced from
hairy roots in MS media supplemented with 1.6 mg/L ZT and 0.4 mg/L NAA. The
PCR-Southern hybridization demonstrated that T-DNA had been integrated into the
chromosome of regenerated plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16989275 [PubMed - indexed for MEDLINE]

1088: Theor Appl Genet. 2006 Nov;113(8):1563-70. Epub 2006 Sep 20.

Complementation of the pina (null) allele with the wild type Pina sequence
restores a soft phenotype in transgenic wheat.

Martin JM, Meyer FD, Smidansky ED, Wanjugi H, Blechl AE, Giroux MJ.

Department of Plant Sciences and Plant Pathology, Montana State University,
Bozeman, MT 59717-3140, USA. jmmartin@montana.edu

The tightly linked puroindoline genes, Pina and Pinb, control grain texture in
wheat, with wild type forms of both giving soft, and a sequence alteration
affecting protein expression or function in either giving rise to hard wheat.
Previous experiments have shown that addition of wild type Pina in the presence
of mutated Pinb gave intermediate grain texture but addition of wild type Pinb
gave soft grain. This raises questions as to whether Pina may be less functional
than Pinb. Our goal here was to develop and characterize wheat lines expressing
the wild type Pina-D1a sequence in hard wheat with the null mutation (Pina-D1b)
for Pina. Three transgenic lines plus Bobwhite were evaluated in two
environments. Grain texture, grain protein, and kernel weight were determined for
the transgenic lines and Bobwhite. The three transgenic lines had soft phenotype,
and none of the transgenic lines differed from Bobwhite for grain protein or
kernel weight. The soft phenotype was accompanied by increases in Pina transcript
accumulation. Total Triton X-114 extractable PINA and PINB increased from 2.5 to
5.5 times those from a soft wheat reference sample, and friabilin, PINA and PINB
bound to starch, increased from 3.8 to 7.8 times those of the soft wheat
reference. Bobwhite showed no starch bound PINA, but transgenic lines had levels
from 5.3 to 13.7 times those of the soft wheat reference sample. Starch bound
PINB in transgenic lines also increased from 0.9 to 2.5 times that for the soft
wheat reference sample. The transgenic expression of wild type Pina sequence in
the Pina null genotype gave soft grain with the characteristics of soft wheat
including increased starch bound friabilin. The results support the hypothesis
that both wild type Pin genes need to be present for friabilin formation and soft
grain.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16988815 [PubMed - indexed for MEDLINE]

1089: Wei Sheng Yan Jiu. 2006 Jul;35(4):431-4.

[Stability of hpt marker gene in transgenic rice in different food matrices and
under varying food-processing conditions]

[Article in Chinese]

Shen LM, Wu YN, Zhang JZ, Zhou PP.

Institute of Nutrition and Food Safety, Chinese Center for Disease Control and
Prevention, Beijing 100050, China.

OBJECTIVE: To estimate the likelihood of horizontal gene transfer from transgenic
rice to bacteria of the food chain and human gut, the stability of s86 transgenic
rice hpt gene in different food matrices and under varying food-processing
conditions was studied. METHODS: Degradation of DNA was monitored by fragment
-multiplex polymerase chain reaction. Integrity of hpt gene in various food
samples was tested. RESULTS: A PCR system for the hpt gene of genetically
modified rice has been established to detect fragments ranging between 236bp and
910bp. Detection of hpt and rbcl gene fragments was carried out in various
food-processed samples by this PCR system. The data showed that the fragments up
to 500 bp were detected in rice and congee, while the fragment length more than
236bp was not detected in crispy rice and popcorn-like rice. CONCLUSION: These
results suggested that there are significant differences in DNA degradation by
different food-processing methods. The likelihood of the large hpt gene fragments
transfer from transgenic rice processed food to bacteria is reduced by food
process.

Publication Types:
English Abstract

PMID: 16986517 [PubMed - in process]

1090: Shokuhin Eiseigaku Zasshi. 2006 Aug;47(4):146-50.

Detection method for genetically modified papaya using duplex PCR.

Yamaguchi A, Shimizu K, Mishima T, Aoki N, Hattori H, Sato H, Ueda N, Watanabe T,
Hino A, Akiyama H, Maitani T.

Japan Food Research Laboratories, Chitose: 2-3, Bunkyo, Chitose-shi, Hokkaido
066-0052, Japan.

A simple and rapid method for the identification of genetically modified (GM)
papaya, derived from Line 55-1, was developed by modifying the Japanese official
PCR method. Genomic DNA was directly extracted from the fresh fruit without the
lyophilization step, using a commercial silica-based kit. To develop a duplex PCR
method which simultaneously detects the GM papaya-specific gene and the intrinsic
papain gene, the papain 2-5'/3' (amplicon size; 184 bp) primer pair for the
detection of the papain gene was newly designed within the region of the products
(211 bp) amplified using the papain 1-5'/-3' primer pair adopted in the Japanese
official PCR method. To detect the GM papaya-specific gene, the primer pair Nos
C-5'/CaM N-3' described in the Japanese official method was used. The DNA
sequences of the GM papaya gene and the intrinsic papain gene were co-amplified
using the PCR method in a single tube. The developed duplex PCR method allows the
simultaneous detection of the products by means of agarose gel electrophoresis or
microchip electrophoresis. The proposed method for GM papaya identification is
simple and rapid.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16984033 [PubMed - indexed for MEDLINE]

1091: Electrophoresis. 2006 Oct;27(20):4029-38.

Quantitative-competitive polymerase chain reaction coupled with slab gel and
capillary electrophoresis for the detection of roundup ready soybean and maize.

Dinelli G, Bonetti A, Marotti I, Minelli M, Navarrete-Casas M, Segura-Carretero
A, Fernö¡ndez-Gutiö©rrez A.

Department of Agro-Environmental Science and Technology, University of Bologna,
Bologna, Italy. gdinelli@agrsci.unibo.it

The aim of the present study was to develop a quantitative-competitive PCR
(QC-PCR) method to detect DNA from transgenic herbicide-resistant (roundup ready,
RR) soybean and maize. Since no QC-PCR system for the quantification of RR maize
had been published at the time of writing, a specific competitor DNA for
transgenic event was developed. For the QC-PCR of RR-soybean, a commercially
available competitor was employed. These internal standards were calibrated by
coamplifying with mixtures containing RR-soybean and maize DNAs. The calibrated
QC-PCR systems were applied to certified RR-soybean and maize flour mixtures in
order to demonstrate their suitability not only for the quantification of the
glyphosate resistance traits in DNA matrices, but also in practically relevant
samples. In addition, a special focus of the present work was to compare the
detection of QC-PCR products by slab gel and CGE with UV detection. CGE permitted
the precise detection of transgenic events also below the equivalence points;
while in slab gel electrophoresis, due to the low sensitivity the quantification
of genetically modified DNA was allowed only at the equivalence point.

PMID: 16983630 [PubMed - indexed for MEDLINE]

1092: Planta. 2007 Mar;225(4):843-62.

The Triticum aestivum non-specific lipid transfer protein (TaLtp) gene family:
comparative promoter activity of six TaLtp genes in transgenic rice.

Boutrot F, Meynard D, Guiderdoni E, Joudrier P, Gautier MF.

INRA, UMR 1096 PIA, 2 place Viala, 34060 Montpellier, France.

Plant non-specific lipid transfer proteins (nsLTPs) are encoded by a multigene
family and support physiological functions, which remain unclear. We adapted an
efficient ligation-mediated polymerase chain reaction (LM-PCR) procedure that
enabled isolation of 22 novel Triticum aestivum nsLtp (TaLtp) genes encoding
types 1 and 2 nsLTPs. A phylogenetic tree clustered the wheat nsLTPs into ten
subfamilies comprising 1-7 members. We also studied the activity of four type 1
and two type 2 TaLtp gene promoters in transgenic rice using the 1-Glucuronidase
reporter gene. The activities of the six promoters displayed both overlapping and
distinct features in rice. In vegetative organs, these promoters were active in
leaves and root vascular tissues while no beta-Glucuronidase (GUS) activity was
detected in stems. In flowers, the GUS activity driven by the TaLtp7.2a,
TaLtp9.1a, TaLtp9.2d, and TaLtp9.3e gene promoters was associated with vascular
tissues in glumes and in the extremities of anther filaments whereas only the
TaLtp9.4a gene promoter was active in anther epidermal cells. In developing
grains, GUS activity and GUS immunolocalization data evidenced complex patterns
of activity of the TaLtp7.1a, TaLtp9.2d, and TaLtp9.4a gene promoters in embryo
scutellum and in the grain epicarp cell layer. In contrast, GUS activity driven
by TaLtp7.2a, TaLtp9.1a, and TaLtp9.3e promoters was restricted to the vascular
bundle of the embryo scutellum. This diversity of TaLtp gene promoter activity
supports the hypothesis that the encoded TaLTPs possess distinct functions in
planta.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16983534 [PubMed - indexed for MEDLINE]

1093: Plant Cell Rep. 2007 Feb;26(2):187-98. Epub 2006 Sep 16.

Agrobacterium tumefaciens mediated transfer of Phaseolus vulgaris alpha-amylase
inhibitor-1 gene into mungbean Vigna radiata (L.) Wilczek using bar as selectable
marker.

Sonia , Saini R, Singh RP, Jaiwal PK.

Advanced Centre for Biotechnology, M D University, Rohtak, India.

Morphologically normal and fertile transgenic plants of mungbean with two
transgenes, bar and alpha-amylase inhibitor, have been developed for the first
time. Cotyledonary node explants were transformed by cocultivation with
Agrobacterium tumefaciens strain EHA105 harboring a binary vector pKSB that
carried bialaphos resistance (bar) gene and Phaseolus vulgaris alpha-amylase
inhibitor-1 (alphaAI-1) gene. Green transformed shoots were regenerated and
rooted on medium containing phosphinothricin (PPT). Preculture and wounding of
the explants, presence of acetosyringone and PPT-based selection of transformants
played significant role in enhancing transformation frequency. Presence and
expression of the bar gene in primary transformants was evidenced by PCR-Southern
analysis and PPT leaf paint assay, respectively. Integration of the Phaseolus
vulgaris alpha-amylase inhibitor gene was confirmed by Southern blot analysis.
PCR analysis revealed inheritance of both the transgenes in most of the T(1)
lines. Tolerance to herbicide was evidenced from seed germination test and
chlorophenol red assay in T(1) plants. Transgenic plants could be recovered after
8-10 weeks of cocultivation with Agrobacterium. An overall transformation
frequency of 1.51% was achieved.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16983450 [PubMed - indexed for MEDLINE]

1094: Regul Toxicol Pharmacol. 2007 Feb;47(1):90-5. Epub 2006 Sep 18.

ELISA method for monitoring human serum IgE specific for Cry1Ab introduced into
genetically modified corn.

Nakajima O, Teshima R, Takagi K, Okunuki H, Sawada J.

National Institute of Health Sciences, Division of Biochemistry and
Immunochemistry, 1-18-1 Kamiyoga, Setagaya, Tokyo, Japan. onakajim@nihs.go.jp

Enzyme-linked immunosorbent assay (ELISA) is the most convenient method of
monitoring the occurrence of IgE antibodies specific for novel proteins in
genetically modified (GM) foods. The levels of IgE specific for a recombinant
protein, Cry1Ab, were determined using an ELISA method. A soluble form of the
Cry1Ab protein purified from pCold1 vector-transformed Escherichia coli
pTf16/BL21 was used as the ELISA coating antigen, and 1M NaCl was used as the
washing buffer to remove IgE non-specifically bound to the coated antigen. Sera
from 44 patients allergic to major food allergens were obtained, diluted 20-fold,
tested, and found no identifiable IgE above background levels. We also tested
sera from patients with corn allergy against whole extracts of non-GM and GM-corn
(MON 810) using immunoblotting. The staining patterns were similar for the two
types of corn. These results indicate that significant levels of IgE antibodies
specific to Cry1Ab were not found in the sera of Japanese patients with food
allergies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16982119 [PubMed - indexed for MEDLINE]

1095: Environ Biosafety Res. 2006 Jan-Mar;5(1):37-46. Epub 2006 Sep 19.

Pyrolysis-field ionization mass spectrometry of rhizodeposits - a new approach to
identify potential effects of genetically modified plants on soil organisms.

Melnitchouck A, Leinweber P, Broer I, Eckhardt KU.

Environmental Solutions, Remediation Services, Calgary, AB T2E 7M8, Canada.

The objectives of the present study were (1) to investigate the qualitative
composition of rhizodeposits leached from soils cropped with non-transgenic and
genetically modified (GM) potatoes, and disclose if there were GM-specific
modifications in potato rhizodeposition, and (2) to compare these results with
conventional bulk parameters of microbial activity in soil. We have raised
potatoes from a non-transgenic line (Solanum tuberosum L. cv. Dö©sirö©e) and three
GM lines, which expressed a gene for the resistance to kanamycin (DLH 9000) and a
gene for T4 lysozyme (DL10 and DL12). A sandy soil placed in 340 cm3-"CombiSart"
containers was used, from which the rhizodeposit was leached after a six-week
growth period. The freeze-dried leachates were analyzed by pyrolysis-field
ionization mass spectrometry (Py-FIMS). The Py-FI mass spectra gave detailed
molecular-chemical information about the composition of leachates, indicating
that the potato growth generally altered the composition of the soil solution.
Moreover, a principal component analysis of the mass spectra showed differences
between the leachates from the non-transgenic parent line and the GM potatoes as
well as among the latter group. However, these differences in molecular
composition could not be assigned to the release of T4-lysozyme into soil.
Dehydrogenase activity and substrate-induced soil respiration as more common bulk
parameters of soil microbial activity failed to disclose any significant effects
of the various potatoes grown. The limitations of the described rhizodeposit
leaching and analysis for risk assessment of GM potato cropping under field
conditions are discussed critically. However, it could be concluded that the
Py-FI mass spectrometric "fingerprint" can be developed as a fast, comprehensive,
highly sensitive and reproducible analytical approach to discern any effects
GM-crops may exert on soil ecological parameters.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 16978573 [PubMed - indexed for MEDLINE]

1096: Environ Biosafety Res. 2006 Jan-Mar;5(1):3-13. Epub 2006 Sep 19.

Hybridization between oilseed rape (Brassica napus) and different populations and
species of Raphanus.

Ammitzbö¸ll H, Bagger Jö¸rgensen R.

Biosystems Department, Risö¸ National Laboratory, 4000 Roskilde, Denmark.

When cultivating genetically modified varieties, the spontaneous gene flow
between crop and wild relatives could be of concern. We analyzed spontaneous
hybridization between a transgenic male-sterile line of oilseed rape (Brassica
napus, 2n = 38, AACC) and, as pollen donors, three European populations of wild
radish (Raphanus raphanistrum, 2n = 18, Rr,Rr) and a variety of cultivated radish
(Raphanus sativus, 2n = 18, RR). Seeds showed size and shape dimorphism that
correlated to the frequency of hybrids. The offspring were scored morphologically
and analyzed using DNA markers (inter-simple sequence repeats) to quantify hybrid
frequencies. Seed set ranged from 0.4-1.2 seeds per pod, and 0.02-0.6 seeds per
pod were confirmed as hybrids. The frequency of confirmed hybrids differed
significantly among populations of R. raphanistrum. In the cross with a French
population, all offspring were hybrids; in the cross with a Swiss population, 53%
of the offspring were hybrids; and in the cross with a Danish population, only 2%
of the offspring were found to be hybrids. The remaining offspring apparently
belonged to two groups: the majority was B. napus-like plants, possibly of
matromorphic origin, and a minority from the Danish cross seemed to carry
fragments of the Raphanus genome. In the cross with a cultivated R. sativus, all
offspring were found to be hybrids. This is the first report on spontaneous
hybridization between B. napus and R. sativus. Hybrids from all
cross-combinations had low pollen fertility (0-15%). If R. raphanistrum occurs
where male-sterile B. napus is cultivated, large regional differences in
hybridization frequencies between the species could complicate environmental risk
assessment of transgenic oilseed rape.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16978570 [PubMed - indexed for MEDLINE]

1097: EMBO Rep. 2006 Oct;7(10):1052-8. Epub 2006 Sep 15.

SERRATE: a new player on the plant microRNA scene.

Lobbes D, Rallapalli G, Schmidt DD, Martin C, Clarke J.

Department of Cell and Developmental Biology, John Innes Centre, Norwich Research
Park, Colney, Norwich NR4 7UH, UK.

MicroRNAs (miRNAs) function as sequence-specific guides that control gene
expression by post-transcriptional gene silencing. Many miRNAs influence plant
development by regulating the accumulation of transcripts that encode
transcription factors. Mutants defective in miRNA accumulation, such as dcl1,
hen1, hyl1 and ago1, have pleiotropic developmental phenotypes. The serrate-1
(se-1) mutant of Arabidopsis also shows a highly pleiotropic phenotype, which
overlaps with the phenotypes of mutants defective in miRNA accumulation. Although
it has been proposed that SERRATE (SE) functions specifically in miRNA-mediated
repression of the leaf polarity genes PHABULOSA and PHAVOLUTA, microarray
analysis shows upregulation of many genes known to be the targets of miRNAs in
se-1. We show that SE is a general regulator of miRNA levels affecting the
processing of primary miRNA to miRNA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16977334 [PubMed - indexed for MEDLINE]

1098: Plant J. 2006 Oct;48(1):125-37.

Cross-talk between ethylene and drought signalling pathways is mediated by the
sunflower Hahb-4 transcription factor.

Manavella PA, Arce AL, Dezar CA, Bitton F, Renou JP, Crespi M, Chan RL.

Cö¡tedra de Biologö­a Celular y Molecular, Facultad de Bioquö­mica y Ciencias
Biolö¨gicas, Universidad Nacional del Litoral, CONICET, CC 242 Ciudad
Universitaria 3000, Santa Fe, Argentina.

Hahb-4 is a member of the Helianthusannuus (sunflower) subfamily I of HD-Zip
proteins that is transcriptionally regulated by water availability and abscisic
acid. Transgenic Arabidopsis thaliana plants overexpressing this transcription
factor (TF) exhibit a characteristic phenotype that includes a strong tolerance
to water stress. Here we show that this TF is a new component of ethylene
signalling pathways, and that it induces a marked delay in senescence. Plants
overexpressing Hahb-4 are less sensitive to external ethylene, enter the
senescence pathway later and do not show the typical triple response.
Furthermore, transgenic plants expressing this gene under the control of its own
inducible promoter showed an inverse correlation between ethylene sensitivity and
Hahb-4 levels. Potential targets of Hahb-4 were identified by comparing the
transcriptome of Hahb-4-transformed and wild-type plants using microarrays and
quantitative RT-PCR. Expression of this TF has a major repressive effect on genes
related to ethylene synthesis, such as ACO and SAM, and on genes related to
ethylene signalling, such as ERF2 and ERF5. Expression studies in sunflower
indicate that Hahb-4 transcript levels are elevated in mature/senescent leaves.
Expression of Hahb-4 is induced by ethylene, concomitantly with several genes
homologous to the targets identified in the transcriptome analysis (HA-ACOa and
HA-ACOb). Transient transformation of sunflower leaves demonstrated the action of
Hahb-4 in the regulation of ethylene-related genes. We propose that Hahb-4 is
involved in a novel conserved mechanism related to ethylene-mediated senescence
that functions to improve desiccation tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16972869 [PubMed - indexed for MEDLINE]

1099: Plant J. 2006 Oct;48(1):113-24.

Multi-site genetic modulation of monolignol biosynthesis suggests new routes for
formation of syringyl lignin and wall-bound ferulic acid in alfalfa (Medicago
sativa L.).

Chen F, Srinivasa Reddy MS, Temple S, Jackson L, Shadle G, Dixon RA.

Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway,
Ardmore, Oklahoma 73401, USA.

Genes encoding seven enzymes of the monolignol pathway were independently
downregulated in alfalfa (Medicago sativa) using antisense and/or RNA
interference. In each case, total flux into lignin was reduced, with the largest
effects arising from the downregulation of earlier enzymes in the pathway. The
downregulation of l-phenylalanine ammonia-lyase, 4-coumarate 3-hydroxylase,
hydroxycinnamoyl CoA quinate/shikimate hydroxycinnamoyl transferase, ferulate
5-hydroxylase or caffeic acid 3-O-methyltransferase resulted in compositional
changes in lignin and wall-bound hydroxycinnamic acids consistent with the
current models of the monolignol pathway. However, downregulating caffeoyl CoA
3-O-methyltransferase neither reduced syringyl (S) lignin units nor wall-bound
ferulate, inconsistent with a role for this enzyme in 3-O-methylation ofS
monolignol precursors and hydroxycinnamic acids. Paradoxically, lignin
composition differed in plants downregulated in either cinnamate 4-hydroxylase or
phenylalanine ammonia-lyase. No changes in the levels of acylated flavonoids were
observed in the various transgenic lines. The current model for monolignol and
ferulate biosynthesis appears to be an over-simplification, at least in alfalfa,
and additional enzymes may be needed for the 3-O-methylation reactions of S
lignin and ferulate biosynthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16972868 [PubMed - indexed for MEDLINE]

1100: Electrophoresis. 2006 Oct;27(19):3879-88.

A new PCR-CGE (size and color) method for simultaneous detection of genetically
modified maize events.

Nadal A, Coll A, La Paz JL, Esteve T, Pla M.

Institut de Tecnologia Agroalimentö ria, Universitat de Girona, EPS, Girona,
Spain.

We present a novel multiplex PCR assay for simultaneous detection of multiple
transgenic events in maize. Initially, five PCR primers pairs specific to events
Bt11, GA21, MON810, and NK603, and Zea mays L. (alcohol dehydrogenase) were
included. The event specificity was based on amplification of transgene/plant
genome flanking regions, i.e., the same targets as for validated real-time PCR
assays. These short and similarly sized amplicons were selected to achieve high
and similar amplification efficiency for all targets; however, its unambiguous
identification was a technical challenge. We achieved a clear distinction by a
novel CGE approach that combined the identification by size and color (CGE-SC).
In one single step, all five targets were amplified and specifically labeled with
three different fluorescent dyes. The assay was specific and displayed an LOD of
0.1% of each genetically modified organism (GMO). Therefore, it was adequate to
fulfill legal thresholds established, e.g., in the European Union. Our CGE-SC
based strategy in combination with an adequate labeling design has the potential
to simultaneously detect higher numbers of targets. As an example, we present the
detection of up to eight targets in a single run. Multiplex PCR-CGE-SC only
requires a conventional sequencer device and enables automation and high
throughput. In addition, it proved to be transferable to a different laboratory.
The number of authorized GMO events is rapidly growing; and the acreage of
genetically modified (GM) varieties cultivated and commercialized worldwide is
rapidly increasing. In this context, our multiplex PCR-CGE-SC can be suitable for
screening GM contents in food.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16972302 [PubMed - indexed for MEDLINE]

1101: J Agric Food Chem. 2006 Sep 20;54(19):7187-92.

Glyphosate-tolerant alfalfa is compositionally equivalent to conventional alfalfa
(Medicago sativa L.).

McCann MC, Rogan GJ, Fitzpatrick S, Trujillo WA, Sorbet R, Hartnell GF, Riodan
SG, Nemeth MA.

Monsanto Company, 800 North Lindbergh Boulevard, St. Louis, Missouri 63167, USA.
melinda.c.mccann@monsanto.com

Glyphosate-tolerant alfalfa (GTA) was developed to withstand over-the-top
applications of glyphosate, the active ingredient in Roundup agricultural
herbicides. As a part of the safety assessment, GTA (designated J101 x J163) was
grown under controlled field conditions at geographically diverse locations
within the United States during the 2001 and 2003 field seasons along with
control and other conventional alfalfa varieties for compositional assessment.
Field trials were conducted using a randomized complete block design with four
replication blocks at each site. Alfalfa forage was harvested at the late bud to
early bloom stage from each plot at five field sites in 2001 (establishment year)
and from four field sites in 2003 (third year of stand). The concentration of
proximate constituents, fibers, amino acids, coumestrol, and minerals in the
forage was measured. The results showed that the forage from GTA J101 x J163 is
compositionally equivalent to forage from the control and conventional alfalfa
varieties.

PMID: 16968081 [PubMed - indexed for MEDLINE]

1102: J Biol Chem. 2006 Nov 10;281(45):34421-9. Epub 2006 Sep 11.

Functional and biochemical analysis of the N-terminal domain of phytochrome A.

Mateos JL, Luppi JP, Ogorodnikova OB, Sineshchekov VA, Yanovsky MJ, Braslavsky
SE, Gö¤rtner W, Casal JJ.

Max-Planck-Institut fö¼r Bioanorganische Chemie, Postfach 101356, D-45413 Mö¼lheim
an der Ruhr, Germany.

Phytochrome A (phyA) is a versatile plant photoreceptor that mediates responses
to brief light exposures (very low fluence responses, VLFR) as well as to
prolonged irradiation (high irradiance responses, HIR). We identified the
phyA-303 mutant allele of Arabidopsis thaliana bearing an R384K substitution in
the GAF subdomain of the N-terminal half of phyA. phyA-303 showed reduced phyA
spectral activity, almost normal VLFR, and severely impaired HIR. Recombinant
N-terminal half oat of PHYA bearing the phyA-303 mutation showed poor
incorporation of chromophore in vitro, despite the predicted relatively long
distance (>13 A) between the mutation and the closest ring of the chromophore.
Fusion proteins bearing the N-terminal domain of oat phyA, beta-glucuronidase,
green fluorescent protein, and a nuclear localization signal showed physiological
activity in darkness and mediated VLFR but not HIR. At equal protein levels, the
phyA-303 mutation caused slightly less activity than the fusions containing the
wild-type sequence. Taken together, these studies highlight the role of the
N-terminal domain of phyA in signaling and of distant residues of the GAF
subdomain in the regulation of phytochrome bilin-lyase activity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16966335 [PubMed - indexed for MEDLINE]

1103: J Neuroendocrinol. 2006 Oct;18(10):776-85.

Exaggerated response of arginine vasopressin-enhanced green fluorescent protein
fusion gene to salt loading without disturbance of body fluid homeostasis in
rats.

Fujio T, Fujihara H, Shibata M, Yamada S, Onaka T, Tanaka K, Morita H, Dayanithi
G, Kawata M, Murphy D, Ueta Y.

Department of Occupational Health, Matsushita Science Center of Industrial
Hygiene, Kadoma, Japan.

We examined the effects of chronic salt loading on the hypothalamic expressions
of the enhanced green fluorescent protein (eGFP), arginine vasopressin (AVP) and
oxytocin (OXT) genes in AVP-eGFP transgenic rats that expressed eGFP in the
hypothalamic AVP-containing neurones. In these rats, salt loading for 5 days
caused a marked increase of the eGFP fluorescence in the magnocellular divisions
of the paraventricular nucleus (PVN), the supraoptic nucleus (SON) and the
internal layer of the median eminence. Expression of the eGFP gene was increased
seven- to eight-fold in the PVN and SON of salt-loaded rats in comparison with
euhydrated rats. By contrast, none of these changes were observed in the
suprachiasmatic nucleus. The expression of the AVP and OXT genes was increased
1.5- to two-fold in the PVN and SON of salt-loaded nontransgenic (control) and
transgenic rats. There were no differences in the expression levels of the AVP
and OXT genes in the PVN and SON between nontransgenic (control) and transgenic
animals under normal conditions and after salt loading. In the posterior
pituitary gland, the intensity of the eGFP fluorescence did not change after salt
loading for 5 days, but increased after 10 days of salt loading. Upon salt
loading, significant increases in the plasma AVP concentrations, plasma
osmolality and plasma Na+ were observed. Furthermore, there were no significant
differences in changes of water intake, food intake, urine volume, urine
osmolality, urine Na+ concentrations, and the body weights in both models under
normal or salt-loaded conditions. Our results show that the response of the
AVP-eGFP fusion gene to chronic salt loading is exaggerated, and humoral
responses such as AVP and OXT and the body fluid homeostasis are maintained in
AVP-eGFP transgenic rats. The AVP-eGFP transgenic rat gives us a new opportunity
to study the dynamics of the AVP system in vivo.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16965296 [PubMed - indexed for MEDLINE]

1104: J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2006
Dec;192(12):1335-48. Epub 2006 Sep 9.

Olfactory conditioning of proboscis activity in Drosophila melanogaster.

Chabaud MA, Devaud JM, Pham-Delö¨gue MH, Preat T, Kaiser L.

Dö©veloppement, Evolution et Plasticitö© du Systö¨me Nerveux, CNRS, Bö¢t. 32/33,
Avenue de la Terrasse, 91198, Gif-sur-Yvette cedex, France.

Olfactory learning and memory processes in Drosophila have been well investigated
with aversive conditioning, but appetitive conditioning has rarely been
documented. Here, we report for the first time individual olfactory conditioning
of proboscis activity in restrained Drosophila melanogaster. The protocol was
adapted from those developed for proboscis extension conditioning in the honeybee
Apis mellifera. After establishing a scale of small proboscis movements necessary
to characterize responses to olfactory stimulation, we applied Pavlovian
conditioning, with five trials consisting of paired presentation of a banana
odour and a sucrose reward. Drosophila showed conditioned proboscis activity to
the odour, with a twofold increase of percentage of responses after the first
trial. No change occurred in flies experiencing unpaired presentations of the
stimuli, confirming an associative basis for this form of olfactory learning. The
adenylyl cyclase mutant rutabaga did not exhibit learning in this paradigm. This
protocol generated at least a short-term memory of 15 min, but no significant
associative memory was detected at 1 h. We also showed that learning performance
was dependent on food motivation, by comparing flies subjected to different
starvation regimes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16964495 [PubMed - indexed for MEDLINE]

1105: Nat Biotechnol. 2006 Sep;24(9):1075-7.

Why spurning food biotech has become a liability.

Miller HI, Conko G, Kershen DL.

Hoover Institution, Stanford University, Stanford, California 94305-6010, USA.
miller@hoover.stanford.edu

PMID: 16964211 [PubMed - indexed for MEDLINE]

1106: Plant J. 2006 Sep;47(6):969-76.

Early infection of scutellum tissue with Agrobacterium allows high-speed
transformation of rice.

Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H.

Plant Genetic Engineering Research Unit, National Institute of Agrobiological
Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan. stoki@affrc.go.jp

Several approaches have recently been adopted to improve Agrobacterium-mediated
transformation of rice, both to generate the large number of T-DNA insertion
plants needed for functional analysis of the rice genome, and for production of
rice with additional agronomical value. However, about 3 months of in vitro
culture is still required for isolation of transgenic rice plants. Here, we
report the competency of scutellum tissue from 1-day pre-cultured seeds for
Agrobacterium-mediated transformation. Furthermore, early infection of rice seeds
with Agrobacterium enhanced efficient selection of transformed calli. Using our
system, we successfully regenerated transgenic rice plantlets within a month of
the start of the aseptic culture of mature seeds. Our new system should reduce
the somaclonal variation accompanying prolonged culture of rice cells in the
dedifferentiated state and facilitate the molecular breeding of rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16961734 [PubMed - indexed for MEDLINE]

1107: Am J Clin Nutr. 2006 Sep;84(3):497-504.

Novel soybean oils with different fatty acid profiles alter cardiovascular
disease risk factors in moderately hyperlipidemic subjects.

Lichtenstein AH, Matthan NR, Jalbert SM, Resteghini NA, Schaefer EJ, Ausman LM.

Cardiovascular Nutrition Laboratory and the Lipid Metabolism Laboratory, Jean
Mayer US Department of Agriculture Human Nutrition Research Center on Aging,
Tufts University, Boston, MA 02111, USA. alice.lichtenstein@tufts.edu

BACKGROUND: A variety of soybean oils were developed with improved oxidative
stability and functional characteristics for use as alternatives to partially
hydrogenated fat. OBJECTIVE: The objective was to assess the effect of
selectively bred and genetically modified soybean oils with altered fatty acid
profiles, relative to common soybean and partially hydrogenated soybean oils, on
cardiovascular disease risk factors. DESIGN: Thirty subjects (16 women and 14
men) aged >50 y with LDL-cholesterol concentrations >130 mg/dL at screening
consumed 5 experimental diets in random order for 35 d each. Diets contained the
same foods and provided 30% of energy as fat, of which two-thirds was either
soybean oil (SO), low-saturated fatty acid soybean oil (LoSFA-SO), high-oleic
acid soybean oil (HiOleic-SO), low-alpha-linolenic acid soybean oil (LoALA-SO),
or partially hydrogenated soybean oil (Hydrog-SO). RESULTS: Plasma phospholipid
patterns reflected the predominant fat in the diet. LDL-cholesterol
concentrations were 3.66 +/- 0.67(b), 3.53 +/- 0.77(b), 3.70 +/- 0.66(b), 3.71
+/- 0.64(a,b), and 3.92 +/- 0.70(a) mol/L; HDL-cholesterol concentrations were
1.32 +/- 0.32(a,b), 1.32 +/- 0.35(b), 1.36 +/- 0.33(a), 1.32 +/- 0.33(b), and
1.32 +/- 0.32(a,b) mol/L for the SO, LoSFA-SO, HiOleic-SO, LoALA-SO, and
Hydrog-SO diets, respectively (values with different superscript letters are
significantly different, P < 0.05). No significant effects were observed on
VLDL-cholesterol, triacylglycerol, lipoprotein(a), and C-reactive protein
concentrations or on ratios of LDL cholesterol to apolipoprotein B (apo B) and
HDL cholesterol to apo A-I. Total cholesterol:HDL cholesterol was lower after
subjects consumed the unhydrogenated soybean oils than after they consumed the
Hydrog-SO diet. CONCLUSIONS: All varieties of soybean oils resulted in more
favorable lipoprotein profiles than did the partially hydrogenated form. These
soybean oils may provide a viable option for reformulation of products to reduce
the content of trans fatty acids.

Publication Types:
Randomized Controlled Trial
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16960162 [PubMed - indexed for MEDLINE]

1108: J Exp Bot. 2006;57(14):3639-45. Epub 2006 Sep 6.

Ketocarotenoid formation in transgenic potato.

Gerjets T, Sandmann G.

Molecular Biosciences 213, J.W. Goethe Universitö¤t, PO Box 111932, D-60054
Frankfurt/M., Germany.

Potato has been genetically engineered for the production of commercially
important ketocarotenoids including astaxanthin (3,3'-dihydroxy
4,4'-diketo-beta-carotene). To support the formation of 3-hydroxylated and
4-ketolated beta-carotene, a transgenic potato line accumulating zeaxanthin due
to inactivated zeaxanthin epoxidase was co-transformed with the crtO
beta-carotene ketolase gene from the cyanobacterium Synechocystis under a
constitutive promoter. Plants were generated which exhibited expression of this
gene, resulting in an accumulation of echinenone, 3'-hydroxyechinenone, and
4-ketozeaxanthin in leaves, as well as 3'-hydroxyechinenone, 4-ketozeaxanthin
together with astaxanthin in the tuber. The amount of ketocarotenoids formed
represent approximately 10-12% of total carotenoids in leaves and tubers.
Negative effects on photosynthesis due to the presence of the ketocarotenoids in
leaves could be excluded by the determination of variable fluorescence.

PMID: 16957020 [PubMed - indexed for MEDLINE]

1109: J Exp Bot. 2006;57(14):3575-82. Epub 2006 Sep 6.

Expression of BjMT2, a metallothionein 2 from Brassica juncea, increases copper
and cadmium tolerance in Escherichia coli and Arabidopsis thaliana, but inhibits
root elongation in Arabidopsis thaliana seedlings.

Zhigang A, Cuijie L, Yuangang Z, Yejie D, Wachter A, Gromes R, Rausch T.

Northeast Forestry, University Key Laboratory of Forest Plant Ecology, Ministry
of Education, Hexing Road 26, Harbin 150040, PR China. zan@hip.uni-hd.de

The protective function of a plant type-2 metallothionein was analysed after
expression in Escherichia coli and in Arabidopsis thaliana seedlings. BjMT2 from
Brassica juncea was expressed in E. coli as a TrxABjMT2 fusion protein. After
affinity chromatography and cleavage from the TrxA domain, pure BjMT2 protein was
obtained which strongly reacted with the thiol reagent monobromobimane.
Escherichia coli cells expressing the TrxABjMT2 fusion were more tolerant to Cu2+
and Cd2+ exposure than control strains. Likewise, when BjMT2 cDNA was expressed
in A. thaliana under the regulation of the 35S promoter, seedlings exhibited an
increased tolerance against Cu2+ and Cd2+ based on shoot growth and chlorophyll
content. Analysis of transiently transformed cells of A. thaliana and tobacco
leaves by confocal laser scanning microscopy (CLSM) revealed exclusive cytosolic
localization of a BjMT2::EGFP (enhanced green fluorescent protein) fusion protein
in control and heavy metal-exposed plant cells. Remarkably, ectopic expression of
BjMT2 reduced root growth in the absence of heavy metal exposure, whereas in the
presence of 50 or 100 microM Cu2+ root growth in control and transgenic lines was
identical. The results indicate that in A. thaliana, root and shoot development
are differentially affected by ectopic expression of BjMT2.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16957018 [PubMed - indexed for MEDLINE]

1110: Virology. 2006 Dec 5-20;356(1-2):171-8. Epub 2006 Sep 7.

Immunogenicity of a plant-derived edible rotavirus subunit vaccine transformed
over fifty generations.

Li JT, Fei L, Mou ZR, Wei J, Tang Y, He HY, Wang L, Wu YZ.

Institute of Immunology, PLA, The Third Military Medical University, Shapingba
District, Chongqing 400038, China.

Major efforts have been put forth for the development of effective rotavirus
vaccines including transgenic plant vaccines. Previous studies have reported that
rotavirus VP7 maintains its neutralizing immunity when it is transformed into the
potato genome. The present study was aimed at investigating the hereditary
stability of VP7-transformed potatoes over fifty generations. The VP7 gene was
stably transcribed and expressed in potato cells as detected by RT-PCR and
Western blotting. Humeral and mucosal responses were successfully induced in
BALB/c mice fed with the fiftieth generation transformed potato tubers. There
were no significant differences in serum IgG and fecal IgA between the mice fed
with the first and fiftieth generation potatoes (P>0.05). Profiles of cytokines
such as IFN-gamma, IL-2, IL-4, IL-5 and TGF-beta in immunized mice showed a naive
T-cells bias to Th1 and Th3 polarization. Moreover, specific CTL responses were
also detected in C57BL/6 mice fed with transformed potatoes. This research
represents a significant step towards the development of rotavirus vaccines
derived from a transgenic plant that can be obtained by long-term and large-scale
vegetative reproduction. To our knowledge, this is the first finding regarding
vaccines derived from plants that can be propagated for many generations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16956640 [PubMed - indexed for MEDLINE]

1111: Curr Biol. 2006 Aug 8;16(15):R563-4.

GMOs still rankle in Europe.

Williamson N.

Publication Types:
News

PMID: 16953534 [PubMed - indexed for MEDLINE]

1112: Planta. 2006 Dec;225(1):75-87. Epub 2006 Sep 5.

Structural and enzymatic characterization of the isoamylase1 homo-oligomer and
the isoamylase1-isoamylase2 hetero-oligomer from rice endosperm.

Utsumi Y, Nakamura Y.

Department of Biological Production, Akita Prefectural University, 241-7
Kaidobata-Nishi, Shimoshinjyo-Nakano, Akita-city, 010-0195, Japan.

The present study established that there are two distinct polymeric forms of
isoamylase1 (ISA1) in rice endosperm: presumably a homo-pentamer of ISA1 and a
hetero-hexamer composed of five ISA1 and one ISA2. The molecular sizes of the
homo- and hetero-oligomers, which could be fractionated by hydrophobic
chromatography, were approximately 420-480 and 510-550 kDa, respectively. The
hetero-oligomer exhibited higher affinities for various branched polyglucans,
especially for phytoglycogen, which had a K(m) value that was approximately 12
times lower relative to that with the homo-oligomer, although no marked
differences were found in chain preferences for debranching of amylopectin and
phytoglycogen between these forms. The hetero-oligomer was active even when
incubated at 50 degrees C for 10 min, while the homo-multimer was completely
inactivated at 40 degrees C in 10 min. When the ISA1 homo-oligomer was incubated
with the ISA2 protein expressed in Escherichia coli and applied onto a
nondenature polyacrylamide gel, additional debranching activity bands which were
specific for the purified ISA1-ISA2 preparation were also detected, indicating
that ISA1 and ISA2 combine to form a hetero-oligomer. These results suggest that
the hetero-oligomer plays a predominant role in the amylopectin biosynthesis in
rice endosperm although the homo-oligomer can complement the function of the
hetero-oligomer at least to some extent.

PMID: 16953433 [PubMed - indexed for MEDLINE]

1113: Theor Appl Genet. 2006 Nov;113(8):1497-504. Epub 2006 Sep 5.

Pm34: a new powdery mildew resistance gene transferred from Aegilops tauschii
Coss. to common wheat (Triticum aestivum L.).

Miranda LM, Murphy JP, Marshall D, Leath S.

Department of Crop Science, North Carolina State University, Box 7629, Raleigh,
NC 27695, USA.

Powdery mildew is a major fungal disease in wheat growing areas worldwide. A
novel source of resistance to wheat powdery mildew present in the germplasm line
NC97BGTD7 was genetically characterized as a monogenic trait in greenhouse and
field trials using F(2) derived lines from a NC97BGTD7 X Saluda cross.
Microsatellite markers were used to map and tag this resistance gene, now
designated Pm34. Three co-dominant microsatellite markers linked to Pm34 were
identified and their most likely order was established as: Xbarc177-5D, 5.4cM,
Pm34, 2.6cM, Xbarc144-5D, 14cM, Xgwm272-5D. These microsatellite markers were
previously mapped to the long arm of the 5D chromosome and their positions were
confirmed using Chinese Spring nullitetrasomic Nulli5D-tetra5A and ditelosomic
Dt5DL lines. Pm2, the only other known Pm gene on chromosome 5D, has been mapped
to the short arm and its specificity is different from that of Pm34.

PMID: 16953419 [PubMed - indexed for MEDLINE]

1114: Mol Cells. 2006 Aug 31;22(1):89-96.

Molecular cloning of a pepper gene that is homologous to SELF-PRUNING.

Kim DH, Han MS, Cho HW, Jo YD, Cho MC, Kim BD.

Center for Plant Molecular Genetics and Breeding Research, Seoul National
University, Seoul 151-921, Korea.

"Determinate" and "indeterminate" inflorescences in plants are controlled by a
single recessive gene, for example, SELF-PRUNING (SP) in Solanum lycopersicum,
TERMINAL FLOWER1 in Arabidopsis, CENTRORADI-ALIS in Antirrhinum, and
CENTRORADIALIS-like gene in tobacco. Pepper (Capsicum annuum L.) is an
indeterminate species in which shoots grow indefinitely. In this study, we cloned
and characterized the pepper SP-like gene (CaSP). RT-PCR revealed that the CaSP
transcript accumulates to higher levels in floral buds than in other organs.
Comparison of genomic DNA and cDNA sequences from indeterminate and determinate
pepper plants revealed the insertion of a single base in the first exon of CaSP
in the determinate pepper plants. CaSP is annotated in linkage group 8
(chromosome 6) of the SNU2 pepper genetic map and showed similar synteny to SP in
tomato. Transgenic tobacco plants overexpressing CaSP displayed late-flowering
phenotypes similar to the phenotypes caused by overexpression of CaSP orthologs
in other plants. Collectively, these results suggest that pepper CaSP is an
ortholog of SP in tomato.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16951555 [PubMed - indexed for MEDLINE]

1115: Genetics. 2006 Nov;174(3):1095-104. Epub 2006 Sep 1.

Trans-kingdom transposition of the maize dissociation element.

Emelyanov A, Gao Y, Naqvi NI, Parinov S.

Temasek Life Sciences Laboratory, The National University of Singapore, 117604
Singapore, Singapore.

Transposons are very valuable tools for genetic manipulation. However, the number
of transposable elements that have been suitably adapted for experimental use is
insufficient and the spectrum of heterologous hosts in which they have been
deployed is restricted. To date, only transposons from animal hosts have been
utilized in heterologous animal species and transposons of plant origin have been
used in plant genetics. There has been no experimental evidence that any of the
known elements could transpose in hosts belonging to both kingdoms. Here we
demonstrate that the maize Dissociation (Ds) element is capable of effective
Activator (Ac) transposase-mediated transposition in the zebrafish Danio rerio,
yielding remarkable germline transmission rates. In addition, mammalian cells
were also found to be conducive to Ds transposition. Furthermore, we demonstrate
that nuclear localization of Ac transposase is essential for genomic Ds
transposition. Our results support the hypothesis that Ac/Ds elements do not rely
on host-specific factors for transposition and that host factors involved in
their mobility mechanism are widely conserved. Finally, even in vertebrate cells,
the Ac/Ds system displays accurate transposition, large-fragment carrying
capacity, high transposition frequencies, efficient germline transmission, and
reporter gene expression, all of which are advantageous for various genetic
applications and animal biotechnology.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16951067 [PubMed - indexed for MEDLINE]

1116: Carcinogenesis. 2007 Feb;28(2):471-8. Epub 2006 Aug 31.

4-monochlorobiphenyl (PCB3) induces mutations in the livers of transgenic Fisher
344 rats.

Lehmann L, L Esch H, A Kirby P, W Robertson L, Ludewig G.

Institute of Applied Biosciences, Section of Food Chemistry and Toxicology,
University of Karlsruhe (TH) Kaiserstrasse 12, D-76131 Karlsruhe, Germany.

4-monochlorobiphenyl (PCB3) is found in small amounts in commercial PCB mixtures,
indoor and outdoor air, and in food. In contrast to highly chlorinated congeners
that are more resistant to metabolic attack, PCB3 is more readily converted by
xenobiotic-metabolizing enzymes to monohydroxy-PCBs and further to
dihydroxy-metabolites, which can be oxidized to quinones. Our recent studies
demonstrated the initiating action of PCB3 in the livers of male rats. Therefore
we hypothesized that PCB3 and/or its metabolite(s) are mutagenic in rat livers in
vivo. To investigate the mutagenicity and the types of mutations generated by
PCB3, male Fischer 344 BigBlue rats, transgenic for the lacI gene, were injected
intraperitoneally with PCB3 (600 micromol/kg), 4-hydroxy-PCB3 (4-HO-PCB3, 400
micromol/kg), 3-methylcholanthrene (3-MC, 300 micromol/kg, positive control) and
corn oil (negative control) once per week, for 4 weeks. Animals were killed 17
days after the last injection and the mutant frequency of the liver lacI gene
determined. 3-MC induced a 4-fold increase of the mutant frequency of the lacI
gene in the liver. The mutant frequency in PCB3-treated animals was also
significantly elevated. In contrast, 4-HO-PCB3 induced a non-significant doubling
of the mutant frequency. The mutation spectrum of solvent control mutants was
characterized by transitions, whereas in 3-MC-animals, transversion and
frameshift mutations predominated. The PCB3-induced mutation spectrum was similar
to that of the 3-MC-induced mutants. In contrast, the mutation spectrum of the
4-HO-PCB3 group hardly differed from that of the control animals. This study
demonstrates for the first time the mutagenicity of a PCB in vivo.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16950798 [PubMed - indexed for MEDLINE]

1117: J Allergy Clin Immunol. 2006 Sep;118(3):711-8. Epub 2006 Jul 12.

Skin prick tests reveal stable and heritable reduction of allergenic potency of
gene-silenced tomato fruits.

Lorenz Y, Enrique E, Lequynh L, Fö¶tisch K, Retzek M, Biemelt S, Sonnewald U,
Vieths S, Scheurer S.

Paul-Ehrlich-Institut, Langen, Germany.

BACKGROUND: Today, for patients with food allergy, the only possibility to
prevent allergic reactions is avoidance of the allergenic food. Genetic
engineering of hypoallergenic plants by means of RNA interference (RNAi) could be
an approach to improve the quality of life of subjects with food allergy.
OBJECTIVES: We sought to achieve stable inhibition of expression of the
allergenic nonspecific lipid transfer protein Lyc e 3 in tomato and to analyze
the reduction of allergenicity in vitro by using histamine release assays and in
vivo by using skin prick tests with transgenic tomato fruits. METHODS: Gene
silencing was performed by means of RNAi and monitored by using Western blotting
with nonspecific lipid transfer protein-specific antibodies and sera from
patients with tomato allergy. Dose-dependent basophil histamine release assays,
prick-to-prick skin testing, and determination of endogenous histamine content
were performed with fruits harvested from plants of the first and second
generation to assess the allergenic potency compared with that of wild-type
fruits. RESULTS: We demonstrated that silencing of Lyc e 3 by means of RNAi
contributes to reduced skin reactivity and is passed on to the next generation of
fruits. A significant reduction of allergenic potency was determined in vitro and
confirmed by using skin prick tests. CONCLUSION: Taken together, these results
indicate that RNAi technology is an effective tool to generate foods with reduced
allergenicity. CLINICAL IMPLICATIONS: Allergen-reduced plant foods might allow
reduction of dietary restrictions for patients allergic to panallergen families.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16950292 [PubMed - indexed for MEDLINE]

1118: Exp Biol Med (Maywood). 2006 Sep;231(8):1346-52.

Accumulation of recombinant SARS-CoV spike protein in plant cytosol and
chloroplasts indicate potential for development of plant-derived oral vaccines.

Li HY, Ramalingam S, Chye ML.

Department of Botany, The University of Hong Kong, Pokfulam Road, Hong Kong,
China.

Plants are promising candidates as bioreactors for the production of oral
recombinant proteins in the biopharmaceutical industry. As an initial step toward
provision of an oral vaccine against the severe acute respiratory syndrome
coronavirus (SARS-CoV), we have expressed a partial spike (S) protein of SARS-CoV
in the cytosol of nuclear-transformed plants and in the chloroplasts of
plastid-transformed plants. In the construction of both nuclear and plastid
transformation vectors, a 2-kilobase nucleotide sequence encoding amino acids
1-658 of the SARS-CoV spike protein (S1) was modified with nucleotide changes,
but not amino acid changes, to optimize codon usage for expression in plants. To
investigate the subcellular localization of S1 during transient expression in
tobacco leaves, a translational fusion consisting of S1 and the green fluorescent
protein (GFP) was generated. Following agroinfiltration of tobacco leaves,
analysis by laser confocal scanning microscopy revealed that the S1:GFP fusion
protein was localized to the cytosol. In stable transgenic tobacco plants and
lettuce plants generated by Agrobacterium-mediated transformation, tobacco and
lettuce leaves were observed to express the S1 at high levels from the
Cauliflower Mosaic Virus 35S promoter with Northern blot analysis. When the S1
was expressed in transplastomic tobacco, S1 messenger RNA and its corresponding
protein were detected on Northern and Western blot analyses, respectively. Our
results demonstrate the feasibility of producing S1 in nuclear- and
chloroplast-transformed plants, indicating its potential in subsequent
development of a plant-derived and safe oral recombinant subunit vaccine against
the SARS-CoV in edible plants.

PMID: 16946403 [PubMed - indexed for MEDLINE]

1119: Vaccine. 2007 Jan 5;25(3):577-84. Epub 2006 Aug 10.

Oral immunogenicity of potato-derived HBsAg middle protein in BALB/c mice.

Youm JW, Won YS, Jeon JH, Ryu CJ, Choi YK, Kim HC, Kim BD, Joung H, Kim HS.

Plant Genomics Research Center, KRIBB, 52 Oun-Dong, Yusung-Gu, Daejon 305-333,
Republic of Korea.

The antibodies to preS2 synthetic peptides have been probed to neutralize
hepatitis B virus (HBV), and also the addition of preS2 sequence could enhance
the antibody response compared with a conventional vaccine in the non- and low
responders. Previously, we generated transgenic potatoes expressing middle
protein, which contains additional 55 amino acid preS2 region at the N-terminus
of the S protein, of HBV to determine the feasibility of developing a
plant-delivered HBV vaccine. In this study, we monitored the immune response
after induction of immunoglobulin by boosting and assessed the efficacy of the
mucosal immune response with regard to generate IgA antibodies. The HBsAg middle
protein expressed in our transgenic potatoes was well immunized at low antigenic
quantities in mice and the induced anti-S or anti-preS2 antibodies were sustained
for the whole period without decrease. Orally delivery of plant-derived HBsAg
middle protein to mice resulted in fecal anti-S or anti-preS2 as well as serum
IgG. In addition, we used antibodies induced from the immunized mice with the
potato-derived rHBsAg in competition assay as competitors to confirm the binding
ability of preS2 antibodies to surface antigen of hepatitis virus. Anti-preS2
antibodies induced from immunized mice with transgenic potatoes effectively
competed with anti-preS2 murine antibody H8 as expected. From these results, the
inclusion of preS2 antigen to HBV plant vaccine may provide additional protective
immunity in the HBV prevention.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16945456 [PubMed - indexed for MEDLINE]

1120: Water Res. 2006 Oct;40(17):3231-8. Epub 2006 Sep 1.

Degradation of plasmid and plant DNA in water microcosms monitored by natural
transformation and real-time polymerase chain reaction (PCR).

Zhu B.

National Water Research Institute, Environment Canada, 11 Innovation Blvd,
Saskatoon, SK, Canada S7N 3H5. bin.zhu@ec.gc.ca

Extracellular DNA exists in the environment and can be taken up by competent
bacterial cells, leading to horizontal gene transfer. The persistence of
extracellular plasmid and plant DNA in water microcosms was monitored in this
study. Water samples were two groundwater (GW1 and GW2) and one river water (RW)
samples. Three treatments included: (1) intact, (2) 0.22 microm
filter-sterilized, and (3) autoclaved water. DNA from a plasmid (pNS1) and a
transgenic Bt (Bacillus thuringiensis) corn line, both carrying a neomycin
phosphotransferase gene (nptII gene) conferring kanamycin and neomycin
resistance, was inoculated into the microcosms at 0.4 and 0.8 microg/ml,
respectively. By monitoring its ability to transform a competent Pseudomonas
stutzeri strain harboring plasmid pMR7 (P. stutzeri pMR7), plasmid DNA was
degraded to undetectable levels in the intact and/or filter-sterilized water
treatments within 48-96 h in GW1, GW2, and RW. Meanwhile, plasmid DNA persisted
in the autoclaved treatment throughout the entire incubation period. For plant
DNA, a highly sensitive real-time PCR method using SYBR Green I was developed to
monitor the degradation dynamics of the nptII gene carried by the transgenic corn
line in the microcosms. The results showed that the concentration of plant DNA
was reduced by two orders of magnitude (from 0.8-0.008 microg/ml) within 96 h in
the intact and filter-sterilized treatments of GW1, GW2, and RW, in contrast to
its persistence in the autoclaved treatment. In addition, no kanamycin resistant
(Km R) transformants were detected from in situ transformation of P. stutzeri
pMR7 with plasmid pNS1 DNA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16945402 [PubMed - indexed for MEDLINE]

1121: BMC Genomics. 2006 Aug 31;7:222.

Complete plastid genome sequence of Daucus carota: implications for biotechnology
and phylogeny of angiosperms.

Ruhlman T, Lee SB, Jansen RK, Hostetler JB, Tallon LJ, Town CD, Daniell H.

Department of Molecular Biology & Microbiology, University of Central Florida,
Biomolecular Science, Building #20, Room 336, Orlando, FL 32816-2364, USA.
daniell@mail.ucf.edu

BACKGROUND: Carrot (Daucus carota) is a major food crop in the US and worldwide.
Its capacity for storage and its lifecycle as a biennial make it an attractive
species for the introduction of foreign genes, especially for oral delivery of
vaccines and other therapeutic proteins. Until recently efforts to express
recombinant proteins in carrot have had limited success in terms of protein
accumulation in the edible tap roots. Plastid genetic engineering offers the
potential to overcome this limitation, as demonstrated by the accumulation of
BADH in chromoplasts of carrot taproots to confer exceedingly high levels of salt
resistance. The complete plastid genome of carrot provides essential information
required for genetic engineering. Additionally, the sequence data add to the
rapidly growing database of plastid genomes for assessing phylogenetic
relationships among angiosperms. RESULTS: The complete carrot plastid genome is
155,911 bp in length, with 115 unique genes and 21 duplicated genes within the
IR. There are four ribosomal RNAs, 30 distinct tRNA genes and 18
intron-containing genes. Repeat analysis reveals 12 direct and 2 inverted repeats
> or = 30 bp with a sequence identity > or = 90%. Phylogenetic analysis of
nucleotide sequences for 61 protein-coding genes using both maximum parsimony
(MP) and maximum likelihood (ML) were performed for 29 angiosperms. Phylogenies
from both methods provide strong support for the monophyly of several major
angiosperm clades, including monocots, eudicots, rosids, asterids, eurosids II,
euasterids I, and euasterids II. CONCLUSION: The carrot plastid genome contains a
number of dispersed direct and inverted repeats scattered throughout coding and
non-coding regions. This is the first sequenced plastid genome of the family
Apiaceae and only the second published genome sequence of the species-rich
euasterid II clade. Both MP and ML trees provide very strong support (100%
bootstrap) for the sister relationship of Daucus with Panax in the euasterid II
clade. These results provide the best taxon sampling of complete chloroplast
genomes and the strongest support yet for the sister relationship of
Caryophyllales to the asterids. The availability of the complete plastid genome
sequence should facilitate improved transformation efficiency and foreign gene
expression in carrot through utilization of endogenous flanking sequences and
regulatory elements.

Publication Types:
Comparative Study
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16945140 [PubMed - indexed for MEDLINE]

1122: Mol Plant Microbe Interact. 2006 Sep;19(9):939-47.

Stage-specific suppression of basal defense discriminates barley plants
containing fast- and delayed-acting Mla powdery mildew resistance alleles.

Caldo RA, Nettleton D, Peng J, Wise RP.

Department of Plant Pathology, Center for Plant Responses to Environmental
Stresses, USDA-ARS, Iowa State University, Ames, IA 50011, USA.

Nonspecific recognition of pathogen-derived general elicitors triggers the first
line of plant basal defense, which in turn, preconditions the host towards
resistance or susceptibility. To elucidate how basal defense responses influence
the onset of Mla (mildew resistance locus a)-specified resistance, we performed a
meta-analysis of GeneChip mRNA expression for 155 basal defense-related genes of
barley (Hordeum vulgare) challenged with Blumeria graminis f. sp. hordei, the
causal agent of powdery mildew disease. In plants containing the fast-acting
Mla1, Mla6, or Mla13 alleles, transcripts hyper-accumulated from 0 to 16 h after
inoculation (hai) in both compatible and incompatible interactions. Suppression
of basal defense-related transcripts was observed after 16 hai only in compatible
interactions, whereas these transcripts were sustained or increased in
incompatible interactions. By contrast, in plants containing wild-type and
mutants of the delayed-acting Mla12 allele, an early hyper-induction of
transcripts from 0 to 8 hai was observed, but the expression of many of these
genes is markedly suppressed from 8 to 16 hai. These results suggest that the
inhibition of basal defense facilitates the development of haustoria by the
pathogen, consequently delaying the onset of host resistance responses. Thus, we
hypothesize that the regulation of basal defense influences host-cell
accessibility to the fungal pathogen and drives allelic diversification of
gene-specific resistance phenotypes.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16941898 [PubMed - indexed for MEDLINE]

1123: Yonsei Med J. 2006 Aug 31;47(4):505-12.

Evaluating the allergic risk of genetically modified soybean.

Kim SH, Kim HM, Ye YM, Kim SH, Nahm DH, Park HS, Ryu SR, Lee BO.

Department of Allergy, Ajou University School of Medicnie, Suwon, Korea.
hspark@ajou.ac.kr

Genetically modified (GM) soybean (carrying the EPSPS transgene) is the most
common GM food in Korea. In order to assess whether genetic modification
increases the allergenic risk of soybeans, the allergenicity and IgE-reactive
components of wild-type and GM soybean extracts were compared in allergic adults
who had been sensitized to soybeans. We enrolled 1,716 adult allergy patients and
40 healthy, non-atopic controls. Skin prick tests and IgE enzyme linked
immunosorbent assays (ELISAs) were performed using wild-type and GM soybean
extracts, along with other common inhaled allergens. The specificities of serum
IgE antibodies from allergic patients and the identities of the IgE-reactive
components of the soybean extracts were compared using ELISA inhibition testing,
2-dimensional gel electrophoresis, and IgE immunoblotting. To evaluate the
effects of digestive enzymes and heat treatment, the soybean extracts were heated
or pre- incubated with or without simulated gastric and intestinal fluids. The
IgE sensitization rates to wild-type and GM soybeans were identical (3.8% of
allergic adults), and circulating IgE antibodies specific for the two extracts
were comparable. The results of the ELISA inhibition test, SDS-PAGE, and IgE
immunoblotting showed a similar composition of IgE-binding components within the
wild-type and GM extracts, which was confirmed using two-dimensional gel
electrophoresis, IgE immunoblotting, and amino acid sequencing. None of the
subjects had a positive response to purified EPSPS protein in the skin prick
test, ELISA, or IgE immunoblot analysis. These findings suggest that the IgE
sensitization rate to GM soybean extracts is identical to that of wild-type
soybean extracts in adult allergy patients. In addition, based on both in vivo
and in vitro methods, the allergenicity of wild type and GM soybean extracts was
identical.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16941740 [PubMed - indexed for MEDLINE]

1124: Horm Metab Res. 2006 Aug;38(8):491-6.

Catalytically inactive lipoprotein lipase overexpression increases insulin
sensitivity in mice.

Shibasaki M, Bujo H, Takahashi K, Murakami K, Unoki H, Saito Y.

Department of Clinical Cell Biology (F5), Graduate School of Medicine, Chiba
University, Chiba, Japan.

Abnormalities in lipoprotein lipase (LPL) function contribute to the development
of hypertriglyceridemia, one of the characteristic disorders observed in the
metabolic syndrome. In addition to the hydrolyzing activity of triglycerides, LPL
modulates various cellular functions via its binding ability to the cell surface.
Here we show the effects of catalytically inactive LPL overexpression on high-fat
diet (HFD)-induced decreased systemic insulin sensitivity in mice. The binding
capacity of catalytically inactive G188E-LPL to C2C12 skeletal muscle cells was
not significantly different from that of wild type LPL. Insulin-stimulated IRS-1
phosphorylation and glucose uptake were increased by addition of wild type or
mutant LPL in C2C12 cells. After 10 weeks' of HFD feeding, mice had significantly
higher blood glucose levels than chow-fed mice in insulin tolerance tests. The
blood glucose levels after insulin injection was significantly decreased in
mutated LPL-overexpressing mice (G188E mice), as well as in wild type
LPL-overexpressing mice (WT mice). Overexpression of catalytically inactive LPL,
as well as wild type LPL, improved impaired insulin sensitivity in mice. These
results show that decreased expression of LPL possibly causes the insulin
resistance, in addition to hypertriglyceridemia, in metabolic syndrome.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16941273 [PubMed - indexed for MEDLINE]

1125: Plant Mol Biol. 2006 Nov;62(4-5):735-52. Epub 2006 Aug 29.

A novel approach for developing resistance in rice against phloem limited viruses
by antagonizing the phloem feeding hemipteran vectors.

Saha P, Dasgupta I, Das S.

Plant Molecular and Cellular Genetics, Bose Institute, P1/12 CIT Scheme VIIM,
Kolkata 700054, India.

Rice production is known to be severely affected by virus transmitting rice
pests, brown planthopper (BPH) and green leafhopper (GLH) of the order hemiptera,
feeding by phloem abstraction. ASAL, a novel lectin from leaves of garlic (Allium
sativum) was previously demonstrated to be toxic towards hemipteran pests when
administered in artificial diet as well as in ASAL expressing transgenic plants.
In this report ASAL was targeted under the control of phloem-specific
Agrobacterium rolC and rice sucrose synthase-1 (RSs1) promoters at the insect
feeding site into popular rice cultivar, susceptible to hemipteran pests. PCR,
Southern blot and C-PRINS analyses of transgenic plants have confirmed stable
T-DNA integration and the transgenes were co-segregated among self-fertilized
progenies. The T(0) and T(1) plants, harbouring single copy of intact T-DNA
expression cassette, exhibit stable expression of ASAL in northern and western
blot analyses. ELISA showed that the level of expressed ASAL was as high as 1.01%
of total soluble protein. Immunohistofluorescence localization of ASAL depicted
the expected expression patterns regulated by each promoter type. In-planta
bioassay studies revealed that transgenic ASAL adversely affect survival, growth
and population of BPH and GLH. GLH resistant T(1) plants were further evaluated
for the incidence of tungro disease, caused by co-infection of GLH vectored Rice
tungro bacilliform virus (RTBV) and Rice tungro spherical virus (RTSV), which
appeared to be dramatically reduced. The result presented here is the first
report of such GLH mediated resistance to infection by RTBV/RTSV in ASAL
expressing transgenic rice plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16941213 [PubMed - indexed for MEDLINE]

1126: Plant Mol Biol. 2006 Dec;62(6):809-23. Epub 2006 Aug 29.

Down-regulation of the maize and Arabidopsis thaliana caffeic acid
O-methyl-transferase genes by two new maize R2R3-MYB transcription factors.

Fornalö© S, Sonbol FM, Maes T, Capellades M, Puigdomö¨nech P, Rigau J,
Caparrö¨s-Ruiz D.

Departament de Genö©tica Molecular, Laboratori de Genö¨tica Molecular Vegetal,
CSIC-IRTA, Jordi Girona 18-26, 08034, Barcelona, Spain.

The maize (Zea mays L.) caffeic acid O-methyl-transferase (COMT) is a key enzyme
in the biosynthesis of lignin. In this work we have characterized the involvement
of COMT in the lignification process through the study of the molecular
mechanisms involved in its regulation. The examination of the maize COMT gene
promoter revealed a putative ACIII box, typically recognized by R2R3-MYB
transcription factors. We used the sequence of known R2R3-MYB factors to isolate
five maize R2R3-MYB factors (ZmMYB2, ZmMYB8, ZmMYB31, ZmMYB39, and ZmMYB42) and
study their possible roles as regulators of the maize COMT gene. The factors
ZmMYB8, ZmMY31, and ZmMYB42 belong to the subgroup 4 of the R2R3-MYB family along
with other factors associated with lignin biosynthesis repression. In addition,
the induction pattern of ZmMYB31 and ZmMYB42 gene expression on wounding is that
expected for repressors of the maize COMT gene. Arabidopsis thaliana plants
over-expressing ZmMYB31 and ZmMYB42 down-regulate both the A. thaliana and the
maize COMT genes. Furthermore, the over-expression of ZmMYB31 and ZmMYB42 also
affect the expression of other genes of the lignin pathway and produces a
decrease in lignin content of the transgenic plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16941210 [PubMed - indexed for MEDLINE]

1127: J Agric Food Chem. 2006 Sep 6;54(18):6527-34.

A real-time quantitative PCR detection method specific to widestrike transgenic
cotton (event 281-24-236/3006-210-23).

Baeumler S, Wulff D, Tagliani L, Song P.

GeneScan Analytics GmbH, Eurofins, Engesserstrasse 4, D-79108 Freiburg, Germany.

In compliance with global regulations on transgenic crops, a real-time
quantitative PCR method specific to Widestrike transgenic cotton (event
281-24-236/3006-210-23, OECD Unique Identifier DAS-24236-5/DAS-21023-5) was
established on the basis of the DNA sequences in the junction between the
transgene insert and cotton genome. The optimized method consists of a DNA
extraction method for cotton seeds and three PCR systems corresponding to a
cotton-specific endogenous reference DNA sequence SAH7 (Sinapis Arabidopsis
Homolog 7) and specific detection of event 281-24-236 and event 3006-210-23. The
method performance including specificity, sensitivity, accuracy, and precision
was determined at a dynamic range of Widestrike DNA levels from 0.04% to 5.0%.
The limits of detection (LOD) and quantification (LOQ) were < or =0.04% and < or
=0.09%, respectively, at 100 ng of DNA sample per reaction. The quantification
results using either the event 281-24-236 or 3006-210-23 system were consistent,
and the relative deviation from the expected (true) value was in the range of
+/-25%. The robustness of the method was demonstrated by a series of tests with
deviations from the optimized assay parameters such as annealing temperature,
extension time, PCR instrument, interlaboratory transferability, etc. All the
measurements from these tests met the criteria set by EU JRC-CRL (European
Commission Joint Research Centre-Community Reference Lab). This real-time
quantitative PCR method is accurate and robust, and is recommended as a global
benchmark method for the detection and quantification of Widestrike cotton. The
method including description, protocol, and performance results is available on
the JRC-CRL website (http://gmo-crl.jrc.it/statusofdoss.htm).

PMID: 16939306 [PubMed - indexed for MEDLINE]

1128: J Econ Entomol. 2006 Aug;99(4):1407-14.

Modeling larval survival and movement to evaluate seed mixtures of transgenic
corn for control of western corn rootworm (Coleoptera: Chrysomelidae).

Onstad DW.

Department of Natural Resources and Environmental Sciences, University of
Illinois, Urbana, IL 61801, USA. onstad@uiuc.edu

I expanded the population dynamics and genetics model published in 2005 by
Crowder and Onstad to include larval survival and movement to evaluate the role
of mixtures of transgenic and nontransgenic corn, Zea mays L., seed for
resistance management of western corn rootworm. I studied both
density-independent and density-dependent toxin survival. In all but the
worst-case scenarios, resistance did not evolve within 30 yr when the resistance
allele, R, was recessive. The standard model with density-independent toxin
survival based on the expression of a medium dose of toxin indicated that 50% R
allele frequency will be reached by years 5 and 7, respectively, with dominant
and partially recessive expression and 20% nontransgenic seed. The standard model
with density-dependent toxin survival indicates that resistance will occur in
year 5 under the same conditions. These results are similar to the published
results of Crowder and Onstad who studied a model with adjacent block refuges and
mostly nonrandom mating in the landscape (random only within each block). Results
depended on the heterozygote advantage (differential survival between SS and RS)
and the degree of random mating provided by the seed mixture.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16937699 [PubMed - indexed for MEDLINE]

1129: J Econ Entomol. 2006 Aug;99(4):1381-7.

Field measures of western corn rootworm (Coleoptera: Chrysomelidae) mortality
caused by Cry34/35Ab1 proteins expressed in maize event 59122 and implications
for trait durability.

Storer NP, Babcock JM, Edwards JM.

Dow AgroSciences LLC, 9930 Zionsville Road, Indianapolis, IN 46268, USA.

Maize, Zea mays L., has been transformed to express the Cry34Ab1 and Cry35Ab1
proteins from Bacillus thuringiensis strain PS149B1. These two proteins act
together as a binary insecticidal protein that is effective against corn rootworm
(Coleoptera: Chrysomelidae) species. The design of the resistance management plan
to preserve the long-term durability of this trait largely depends on the level
of rootworm mortality induced by Cry34/35Ab1 corn rootworm-protected maize
(frequently referred to as "dose" in this context). Here, we report on studies
that showed Cry34/35Ab1-expressing maize event 59122 caused 99.1 to 99.98%
mortality of western corn rootworm, Diabrotica virgifera virgifera LeConte,
larvae, after adjusting adult emergence numbers for density-dependent mortality.
In two of three studies, there was a short delay in time to 50% adult emergence
from 59122 maize plots compared with control plots, although emergence was
completed at approximately the same time from both types of maize. These data
support an expectation that alleles conferring resistance to the Cry34/35Ab1
proteins in western corn rootworm will be functionally nearly completely to
completely recessive on 59122 maize and that there is unlikely to be assortative
mating of Cry34/35Ab1-resistant and susceptible rootworms. When incorporated into
simulation models of rootworm adaptation to transgenic maize, these findings
suggest that a 20% refuge is likely to be highly effective at prolonging the
durability of 59122 maize.

PMID: 16937696 [PubMed - indexed for MEDLINE]

1130: J Econ Entomol. 2006 Aug;99(4):1085-95.

Effect of corn hybrids expressing the coleopteran-specific cry3Bb1 protein for
corn rootworm control on aboveground insect predators.

Ahmad A, Wilde GE, Whitworth RJ, Zolnerowich G.

Department of Entomology, Kansas State University, Manhattan, KS 66506-4004, USA.
aahmad@ksu.edu

Field and laboratory studies were conducted to determine the effect of transgenic
Bacillus thuringiensis (Bt) corn, Zea mays L. (YieldGard Rootworm), expressing
the Cry3Bb1 protein on aboveground nontarget insect predators (minute pirate bug,
ladybird beetles, and carabids). Visual counts of adult and immature Orius
insidiosus (Say), Coleomegilla maculata (DeGeer), Hippodamia convergens
Gurin-Meneville, and Scymnus spp. occurring in Bt corn and its non-Bt isoline
were made at Manhattan, KS, in 2002 and at Manhattan and Scandia, KS, in 2003. No
significant differences were found between the Bt corn and non-Bt isoline plots
in the abundance (number per plant) of O. insidiosus, C. maculata, H. convergens,
and Scymnus spp. Field predation on Ostrinia nubilalis (Hö¼bner) (Lepidoptera:
Crambidae) egg masses was also observed during the silking stage of corn at
Manhattan and Scandia in 2003. No significant differences were observed among
treatments in predation rate for predators with chewing versus sucking
mouthparts. Two laboratory studies determined the effect of Cry3Bb1 protein
expressed in Bt corn pollen on C. maculata and carabids. The larvae of C.
maculata were reared on Bt pollen, non-Bt pollen, or greenbugs, Schizaphis
graminum (Rondani). The duration of larval and pupal stages, developmental time
from egg hatch to adult emergence, percentage of survival, and elytra length were
compared among treatments. There were no significant differences in developmental
time of larvae fed pollen or greenbugs during their first two instars. However,
significantly prolonged development of the third (1 d) and fourth instars (2 d)
was observed for larvae fed greenbugs only. Total time for larval development was
significantly longer for larvae that fed on greenbugs versus larvae fed on
pollen. No significant differences were observed among treatments in the
percentage of larvae that pupated or pupal stage duration. Larvae that fed on
greenbugs had higher pupal and adult weights compared with pollen-fed larvae.
However, pupal and adult weights did not vary between the Bt and non-Bt pollen
treatments. No significant differences occurred in longevity and elytra length of
beetles among all treatments. Two carabid species, Harpalus caliginosus F. and
Harpalus pensylvanicus DeGeer, were reared on moistened dog food sprinkled with
Bt or non-Bt corn pollen. No significant differences in mortality of H.
caliginosus and H. pensylvanicus were detected among any of the treatments. There
was no significant effect of Bt pollen on fecundity and egg viability of H.
caliginosus. Our studies showed that YieldGard Rootworm had no effect on the
selected coleopteran predators; therefore, this Bt corn hybrid could be used in
an integrated pest management system.

PMID: 16937659 [PubMed - indexed for MEDLINE]

1131: Biotechnol Lett. 2006 Oct;28(19):1551-7. Epub 2006 Aug 3.

A multi-needle-assisted transformation of soybean cotyledonary node cells.

Xue RG, Xie HF, Zhang B.

Department of Life and Science, Laiyang Agricultural College, Qingdao 266109,
China. xuerengao@163.com

A new and simple method for wounding cotyledonary node cells of soybean [Glycine
max (L) Merrill] was developed for obtaining a high frequency of transformants.
Soybean seeds were germinated for 1 day, and the cotyledonary node cells of
half-seeds were wounded mechanically by using a multi-needle consisting of thin
30 fibers. The wounded half-seeds were inoculated with Agrobacterium tumefaciens
cells harboring a recombinant DNA that contained the bar and sgfp genes
conferring phosphinothricin (PPT)-resistance and green fluorescent protein (GFP)
activity, respectively. The inoculated explants were selected on medium
containing 5 or 3 mg PPT/l. The transformation efficiency of soybean was up to
12%. Polymerase chain reaction and genomic Southern blot analysis confirmed
stable integration of the transgenes in the genome of the PPT-resistant plants.
GFP analysis revealed that the transgenes were highly expressed in the plantlets.
Adult plants were resistant to 100 mg PPT/l applied on the leaves, demonstrating
their herbicide-resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16937246 [PubMed - indexed for MEDLINE]

1132: Plant Cell Rep. 2007 Jan;26(1):115-24. Epub 2006 Aug 26.

Enhanced stress tolerance in transgenic pine expressing the pepper CaPF1 gene is
associated with the polyamine biosynthesis.

Tang W, Newton RJ, Li C, Charles TM.

Department of Biology, Howell Science Complex, East Carolina University,
Greenville, NC 27858-4353, USA. tangw@mail.ecu.edu

ERF/AP2 transcription factors play an important role in plant stress tolerance.
However, little is known about the functional significance of ERF/AP2 genes in
pine, compared to the model plant species Arabidopsis. Capsicum annuum pathogen
and freezing tolerance-related protein 1 (CaPF1) is an ERF/AP2 transcription
factor. We show here that overexpression of CaPF1 resulted in a dramatic increase
in tolerance to drought, freezing, and salt stress in a gymnosperm species,
eastern white pine (Pinus strobus L.). Measurement of polyamines demonstrated
that the levels of putrescine (Put), spermidine (Spd), and spermine (Spm) did not
increase but remain constant in CaPF1-overexpressed eastern white pine, whereas
the levels decreased in the controls, probably increasing the ability of
transgenic callus cultures and plants to stress tolerance. These results
demonstrated that enhanced stress tolerance in transgenic pine expressing the
pepper CaPF1 gene is associated with the polyamine biosynthesis and this pepper
transcription factor may be used to engineer pine species for multiple stress
tolerance.

PMID: 16937149 [PubMed - indexed for MEDLINE]

1133: Planta. 2007 Feb;225(3):575-88. Epub 2006 Aug 26.

The barley ERF-type transcription factor HvRAF confers enhanced pathogen
resistance and salt tolerance in Arabidopsis.

Jung J, Won SY, Suh SC, Kim H, Wing R, Jeong Y, Hwang I, Kim M.

School of Agricultural Biotechnology, Seoul National University, Seoul 151-921,
Republic of Korea.

We isolated HvRAF (Hordeum vulgare root abundant factor), a cDNA encoding a novel
ethylene response factor (ERF)-type transcription factor, from young seedlings of
barley. In addition to the most highly conserved APETALA2/ERF DNA-binding domain,
the encoded protein contained an N-terminal MCGGAIL signature sequence, a
putative nuclear localization sequence, and a C-terminal acidic transcription
activation domain containing a novel mammalian hemopexin domain signature-like
sequence. Their homologous sequences were found in AAK92635 from rice and RAP2.2
from Arabidopsis; the ERF proteins most closely related to HvRAF, reflecting
their functional importance. RNA blot analyses revealed that HvRAF transcripts
were more abundant in roots than in leaves. HvRAF expression was induced in
barley seedlings by various treatment regimes such as salicylic acid, ethephon,
methyl jasmonate, cellulase, and methyl viologen. In a subcellular localization
assay, the HvRAF-GFP fusion protein was targeted to the nucleus. The fusion
protein of HvRAF with the GAL4 DNA-binding domain strongly activated
transcription in yeast. Various deletion mutants of HvRAF indicated that the
transactivating activity was localized to the acidic domain of the C-terminal
region, and that the hemopexin domain signature-like sequence was important for
the activity. Overexpression of the HvRAF gene in Arabidopsis plants induced the
activation of various stress-responsive genes, including PDF1.2, JR3, PR1, PR5,
KIN2, and GSH1. Furthermore, the transgenic Arabidopsis plants showed enhanced
resistance to Ralstonia solanacearum strain GMI1000, as well as seed germination
and root growth tolerance to high salinity. These results collectively indicate
that HvRAF is a transcription factor that plays dual regulatory roles in response
to biotic and abiotic stresses in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16937017 [PubMed - indexed for MEDLINE]

1134: Plasmid. 2007 Jan;57(1):18-28. Epub 2006 Aug 28.

A role for the tet(O) plasmid in maintaining Campylobacter plasticity.

Friis LM, Pin C, Taylor DE, Pearson BM, Wells JM.

Institute of Food Research, Colney Lane, Norwich, Norfolk NR4 7UA, UK.
lfriis@ualberta.ca

Genomic sequencing projects are beginning to reveal regions of extensive DNA
homology between bacterial genera. Public fears of the spread of genetically
modified organisms into the food chain and the increasing prevalence of
multi-drug resistant disease in humans highlight the implications of horizontal
gene transfer. The striking DNA sequence similarity between the two uniquely
identified tetracycline resistant (Tc(R)) Campylobacter plasmids, pCC31 and pTet,
suggests their conserved acquisition and maintenance within Campylobacter
[Batchelor, R.A., Pearson, B.M., Friis, L.M., Guerry, P., Wells, J.M. 2004.
Nucleotide sequences and comparison of two large conjugative plasmids from
different Campylobacter species. Microbiology 150, 3507-3517]. It is thus likely
that these and other conjugative plasmids are highly prevalent and broadly
distributed across several continents. Microarray technology is now enabling fast
and extensive genomic comparisons to be made and allows us to investigate intra-
and inter-genetic conservation and variability. This study details the
development of a microarray specific for genes from Campylobacter plasmids pCC31,
pTet and pVir and its application to the analysis of Campylobacter plasmid gene
presence and preservation throughout environmental and clinical isolates.
Application of the iterative algorithm GENCOM (freely available at ) is used as a
rapid and effective way of comparing the content and conservation of plasmids in
bacteria and provides details of the Campylobacter flexible gene pool and its
contribution to genomic plasticity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16934869 [PubMed - indexed for MEDLINE]

1135: Plant Biol (Stuttg). 2006 Sep;8(5):662-72. Epub 2006 Aug 24.

Evaluation of a non-targeted "omic" approach in the safety assessment of
genetically modified plants.

Metzdorff SB, Kok EJ, Knuthsen P, Pedersen J.

Danish Institute for Food and Veterinary Research, 19 Mö¸rkhö¸j Bygade, 2860
Sö¸borg, Denmark. metz@dfvf.dk

Genetically modified plants must be approved before release in the European
Union, and the approval is generally based upon a comparison of various
characteristics between the transgenic plant and a conventional counterpart. As a
case study, focusing on safety assessment of genetically modified plants, we here
report the development and characterisation of six independently transformed
ARABIDOPSIS THALIANA lines modified in the flavonoid biosynthesis. Analyses of
integration events and comparative analysis for characterisation of the intended
effects were performed by PCR, quantitative Real-time PCR, and High Performance
Liquid Chromatography. Analysis by cDNA microarray was used as a non-targeted
approach for the identification of potential unintended effects caused by the
transformation. The results revealed that, although the transgenic lines
possessed different types of integration events, no unintended effects were
identified. However, we found that the majority of genes showing differential
expression were identified as stress-related genes and that environmental
conditions had a large impact on the expression of several genes, proteins, and
metabolites. We suggest that the microarray approach has the potential to become
a useful tool for screening of unintended effects, but state that it is crucial
to have substantial information on the natural variation in traditional crops in
order to be able to interpret "omics" data correctly within the framework of food
safety assessment strategies of novel plant varieties, including genetically
modified plant varieties.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16933176 [PubMed - indexed for MEDLINE]

1136: Plant Mol Biol. 2006 Aug;61(6):897-915.

Expression and functional roles of the pepper pathogen-induced transcription
factor RAV1 in bacterial disease resistance, and drought and salt stress
tolerance.

Sohn KH, Lee SC, Jung HW, Hong JK, Hwang BK.

Laboratory of Molecular Plant Pathology, College of Life Sciences and
Biotechnology, Korea University, Seoul, 136-713, Republic of Korea.

A novel pathogen-induced gene encoding the RAV (Related to ABI3/VP1)
transcription factor, CARAV1, was isolated from pepper leaves infected with
Xanthomonas campestris pv. vesicatoria. CARAV1 contains two distinct DNA-binding
domains AP2 and B3 uniquely found in higher plants. Transient expression analysis
of the smGFP:CARAV1 fusion construct in Arabidopsis protoplasts and pepper
epidermal cells revealed the CARAV1 protein to be localized in the nucleus. The
N-terminal region of CARAV1 fused to the GAL4 DNA-binding domain was required to
activate transcription of reporter genes in yeast. In yeast one-hybrid, the
recognition of CAACA and CACCTG motifs also were essential for the CARAV1 protein
to bind to a specific target gene and activate the reporter gene. The expression
of the CARAV1 gene was strongly induced early in pepper leaves during the
pathogen infection, abiotic elicitors and environmental stresses. CARAV1
transcripts were localized in the phloem cells of leaf tissues during pathogen
infection and ethylene treatment. Ectopic expression of the CARAV1 gene in
transgenic Arabidopsis plants induced some PR genes and enhanced resistance
against infection by Pseudomonas syringae pv. tomato DC3000 and osmotic stresses
by high salinity and dehydration. The CARAV1 promoter activation was induced by
P. syringae pv. tabaci, salicylic acid and abscisic acid. These data suggest that
pathogen- and abiotic stress-inducible CARAV1 functions as a transcriptional
activator triggering resistance to bacterial infection and tolerance to osmotic
stresses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16927203 [PubMed - indexed for MEDLINE]

1137: Plant Mol Biol. 2006 Aug;61(6):829-44.

Identification and functional characterization of a leucine-rich repeat
receptor-like kinase gene that is involved in regulation of soybean leaf
senescence.

Li XP, Gan R, Li PL, Ma YY, Zhang LW, Zhang R, Wang Y, Wang NN.

Department of Plant Biology and Ecology, Nankai University, Tianjin, 300071, PR
China.

We report here the cloning and characterization of a soybean receptor-like kinase
(RLK) gene, designated GmSARK (Glycine max senescence-associated receptor-like
kinase), which is involved in regulating leaf senescence. The conceptual protein
product of GmSARK contains typical domains of LRR receptor-like kinases: a
cytoplasmic domain with all the 11 kinase subdomains, a transmembrane domain and
an extracelullar domain containing 9 Leucine-Rich Repeat (LRR) units that may act
as a receptor. The expression of GmSARK in soybean leaves was up-regulated in all
the three tested senescence systems: senescing cotyledons, dark-induced primary
leaf senescence and the natural leaf senescence process after florescence.
Furthermore, the RNA interference (RNAi)-mediated knocking-down of GmSARK
dramatically retarded soybean leaf senescence. A more complex thylakoid membrane
system, higher foliar level of chlorophyll content and a very remarkable delay of
senescence-induced disintegration of chloroplast structure were observed in
GmSARK-RNAi transgenic leaves. A homolog of maize lethal leaf-spot 1 gene, which
has been suggested to encode a key enzyme catalyzing chlorophyll breakdown, was
isolated and nominated Gmlls1. The expression level of Gmgtr1 gene, which encodes
a key enzyme of chlorophyll synthesis, was also analyzed. It was found that
Gmlls1 was up-regulated and Gmgtr1 was down-regulated during senescence in
wild-type soybean leaves. However, both of the up-regulation of Gmlls1 and
down-regulation of Gmgtr1 were retarded during senescence of GmSARK-RNAi
transgenic leaves. In addition, over-expression of the GmSARK gene greatly
accelerated the senescence progression of CaMV 35S:GmSARK transgenic plants.
Taken together, these results strongly suggested the involvement of this LRR-RLK
in regulation of soybean leaf senescence, maybe via regulating chloroplast
development and chlorophyll accumulation. Multiple functions of GmSARK besides
its regulation of leaf senescence were also discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16927199 [PubMed - indexed for MEDLINE]

1138: Mol Cell Proteomics. 2006 Dec;5(12):2228-43. Epub 2006 Aug 22.

Proteomics fingerprinting of phagosome maturation and evidence for the role of a
Galpha during uptake.

Gotthardt D, Blancheteau V, Bosserhoff A, Ruppert T, Delorenzi M, Soldati T.

Department of Molecular Cell Research, Max Planck Institute for Medical Research,
University Hospital of Heidelberg, Germany.

Phagocytosis, whether of food particles in protozoa or bacteria and cell remnants
in the metazoan immune system, is a conserved process. The particles are taken up
into phagosomes, which then undergo complex remodeling of their components,
called maturation. By using two-dimensional gel electrophoresis and mass
spectrometry combined with genomic data, we identified 179 phagosomal proteins in
the amoeba Dictyostelium, including components of signal transduction, membrane
traffic, and the cytoskeleton. By carrying out this proteomics analysis over the
course of maturation, we obtained time profiles for 1,388 spots and thus
generated a dynamic record of phagosomal protein composition. Clustering of the
time profiles revealed five clusters and 24 functional groups that were mapped
onto a flow chart of maturation. Two heterotrimeric G protein subunits, Galpha4
and Gbeta, appeared at the earliest times. We showed that mutations in the genes
encoding these two proteins produce a phagocytic uptake defect in Dictyostelium.
This analysis of phagosome protein dynamics provides a reference point for future
genetic and functional investigations.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 16926386 [PubMed - indexed for MEDLINE]

1139: Plant Cell Physiol. 2006 Sep;47(9):1187-94. Epub 2006 Aug 22.

AtVPS29, a putative component of a retromer complex, is required for the
efficient sorting of seed storage proteins.

Shimada T, Koumoto Y, Li L, Yamazaki M, Kondo M, Nishimura M, Hara-Nishimura I.

Graduate School of Science, Kyoto University, Kyoto, 606-8502 Japan.

Seed storage proteins are synthesized on rough endoplasmic reticulum (ER) as
larger precursors and are sorted to protein storage vacuoles, where they are
converted into the mature forms. We report here an Arabidopsis mutant, maigo 1
(mag1), which abnormally accumulates the precursors of two major storage
proteins, 12S globulin and 2S albumin, in dry seeds. Electron microscopy revealed
that mag1 seeds mis-sort storage proteins by secreting them from cells. mag1
seeds have smaller protein storage vacuoles in the seeds than do wild-type seeds.
The MAG1 gene encodes a homolog of the yeast (Saccharomyces cerevisiae) protein
VPS29. VPS29 is a component of a retromer complex for recycling a vacuolar
sorting receptor VPS10 from the pre-vacuolar compartment to the Golgi complex.
Our findings suggest that MAG1/AtVPS29 protein is involved in recycling a plant
receptor for the efficient sorting of seed storage proteins. The mag1 mutant
exhibits a dwarf phenotype. A plant retromer complex plays a significant role in
plant growth and development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16926167 [PubMed - indexed for MEDLINE]

1140: J Biol Chem. 2006 Oct 27;281(43):32395-402. Epub 2006 Aug 22.

Cloning and characterization of deoxymugineic acid synthase genes from
graminaceous plants.

Bashir K, Inoue H, Nagasaka S, Takahashi M, Nakanishi H, Mori S, Nishizawa NK.

Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1
Yayoi, Bunkyo-ku, Tokyo 113-8657.

Graminaceous plants have evolved a unique mechanism to acquire iron through the
secretion of a family of small molecules, called mugineic acid family
phytosiderophores (MAs). All MAs are synthesized from l-Met, sharing the same
pathway from l-Met to 2'-deoxymugineic acid (DMA). DMA is synthesized through the
reduction of a 3''-keto intermediate by deoxymugineic acid synthase (DMAS). We
have isolated DMAS genes from rice (OsDMAS1), barley (HvDMAS1), wheat (TaD-MAS1),
and maize (ZmDMAS1). Their nucleotide sequences indicate that OsDMAS1 encodes a
predicted polypeptide of 318 amino acids, whereas the other three orthologs all
encode predicted polypeptides of 314 amino acids and are highly homologous
(82-97.5%) to each other. The DMAS proteins belong to the aldo-keto reductase
superfamily 4 (AKR4) but do not fall within the existing subfamilies of AKR4 and
appear to constitute a new subfamily within the AKR4 group. All of the proteins
showed DMA synthesis activity in vitro. Their enzymatic activities were highest
at pH 8-9, consistent with the hypothesis that DMA is synthesized in subcellular
vesicles. Northern blot analysis revealed that the expression of each of the
above DMAS genes is up-regulated under iron-deficient conditions in root tissue,
and that of the genes OsDMAS1 and TaDMAS1 is up-regulated in shoot tissue.
OsDMAS1 promoter-GUS analysis in iron-sufficient roots showed that its expression
is restricted to cells participating in long distance transport and that it is
highly up-regulated in the entire root under iron-deficient conditions. In shoot
tissue, OsDMAS1 promoter drove expression in vascular bundles specifically under
iron-deficient conditions.

Publication Types:
Comparative Study

PMID: 16926158 [PubMed - indexed for MEDLINE]

1141: Sci Am. 2006 Sep;295(3):24.

Old MacDonald's pharm.

Choi CQ.

Publication Types:
News

PMID: 16925026 [PubMed - indexed for MEDLINE]

1142: Planta. 2007 Feb;225(3):523-39. Epub 2006 Aug 22.

Functional characterization of Nicotiana benthamiana homologs of peanut water
deficit-induced genes by virus-induced gene silencing.

Senthil-Kumar M, Govind G, Kang L, Mysore KS, Udayakumar M.

Department of Crop Physiology, University of Agricultural Sciences, GKVK,
Bangalore, 560 065 Karnataka, India.

Determining the functional role of genes that are differentially regulated during
a stress response is challenging. In this study, few water deficit-induced genes
from peanut were characterized in Nicotiana benthamiana using virus-induced gene
silencing (VIGS) and their relevance for stress adaptation was validated.
Twenty-five cDNA clones from peanut water deficit stress-induced cDNA library
that had more than 50% nucleotide similarity with N. benthamiana or tomato
homologs were selected. VIGS in peanut is not yet feasible and therefore we
characterized these 25 genes in N. benthamiana. Increased membrane damage was
seen under water deficit stress in most of the silenced plants signifying that
many of these stress-induced genes are important to confer drought tolerance.
Among the genes tested, silencing by homolog of flavonol 3-O-glucosyltransferase
(F3OGT), homolog of alcohol dehydrogenase, homologous to salt inducible protein,
and homolog of heat shock protein 70 showed more visible wilting symptoms
compared with the control plants during water deficit stress. Interestingly,
down-regulation of two genes, homologous to aspartic proteinase 2, and homolog of
Jumonji class of transcription factor showed relative drought tolerant
phenotypes. F3OGT silenced plants showed more wilting symptoms, membrane damage
and chlorophyll degradation than any other silenced plants during water deficit.
Our results demonstrate that VIGS approach can be used to characterize and assess
the functional relevance of water deficit stress-induced cDNAs in a heterologous
species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16924536 [PubMed - indexed for MEDLINE]

1143: Planta. 2007 Feb;225(3):665-79. Epub 2006 Aug 22.

Modification of phenolic metabolism in soybean hairy roots through down
regulation of chalcone synthase or isoflavone synthase.

Lozovaya VV, Lygin AV, Zernova OV, Ulanov AV, Li S, Hartman GL, Widholm JM.

Department of Crop Sciences, University of Illinois, 1201 W. Gregory Drive,
Urbana, IL 61801, USA. lozovaya@uiuc.edu

Soybean hairy roots, transformed with the soybean chalcone synthase (CHS6) or
isoflavone synthase (IFS2) genes, with dramatically decreased capacity to
synthesize isoflavones were produced to determine what effects these changes
would have on susceptibility to a fungal pathogen. The isoflavone and coumestrol
concentrations were decreased by about 90% in most lines apparently due to gene
silencing. The IFS2 transformed lines had very low IFS enzyme activity in
microsomal fractions as measured by the conversion of naringenin to genistein.
The CHS6 lines with decreased isoflavone concentrations had 5 to 20-fold lower
CHS enzyme activities than the appropriate controls. Both IFS2 and CHS
transformed lines accumulated higher concentrations of both soluble and cell wall
bound phenolic acids compared to controls with higher levels found in the CHS6
lines indicating alterations in the lignin biosynthetic branch of the pathway.
Induction of the soybean phytoalexin glyceollin, of which the precursor is the
isoflavone daidzein, by the fungal pathogen Fusarium solani f. sp. glycines (FSG)
that causes soybean sudden death syndrome (SDS) showed that the low isoflavone
transformed lines did not accumulate glyceollin while the control lines did. The
(iso)liquritigenin content increased upon FSG induction in the IFS2 transformed
roots indicating that the pathway reactions before this point can control
isoflavonoid synthesis. The lowest fungal growth rate on hairy roots was found on
the FSG partially resistant control roots followed by the SDS sensitive control
roots and the low isoflavone transformants. The results indicate the importance
of phytoalexin synthesis in root resistance to the pathogen.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16924535 [PubMed - indexed for MEDLINE]

1144: Microb Ecol. 2006 Oct;52(3):583-95. Epub 2006 Aug 19.

Long-term field release of bioluminescent Sinorhizobium meliloti strains to
assess the influence of a recA mutation on the strains' survival.

Selbitschka W, Keller M, Miethling-Graff R, Dresing U, Schwieger F, Krahn I,
Homann I, Dammann-Kalinowski T, Pö¼hler A, Tebbe CC.

Lehrstuhl fö¼r Genetik, Universitö¤t Bielefeld, Postfach 100131, D-33501 Bielefeld,
Germany. Werner.Selbitschka@Genetik.Uni-Bielefeld.DE

A field release experiment was carried out to study the fate of the isogenic,
firefly luciferase (luc) gene-tagged Sinorhizobium meliloti strains L1 (RecA-)
and L33 (RecA+) in the environment. Both strains were released at concentrations
of approximately 10(6) cfu g(-1) soil in replicate and randomized field plots,
which had been sown with alfalfa (Medicago sativa). The survival of both strains
during the following 7 years could be subdivided into three phases: a sharp
decline for more than two orders of magnitude within the first 4 months (phase
I), followed by fluctuations around an average number of 10(4) cfu g(-1) soil for
nearly 4 years (phase II), and a further decline to approximately 60 cfu g(-1)
(phase III). At most sampling dates, no significant differences in the survival
of both strains were detected, indicating that the recA gene function was
dispensable under these environmental conditions. During the field inoculation,
both strains were dispersed accidentally by wind in small numbers to
noninoculated field plots. Strain L33 established at a concentration of more than
10(3) cfu g(-1) soil with subsequent seasonal fluctuations. Although strain L1
must have been disseminated to a similar extent, it could never be recovered from
noninoculated field plots, indicating that the recA mutation interfered with the
strain's capability to establish there. At the beginning of the field experiment,
an indigenous alfalfa-nodulating population was below the limit of detection. In
the following years, however, an indigenous population arose, which finally
outcompeted both strains for saprophytic growth and alfalfa nodulation. RecA-
strain L1 was outcompeted for alfalfa nodulation slightly faster than its RecA+
counterpart L33. The diversity of the indigenous population was characterized by
employing the Enterobacterial Repetitive Intergenic Consensus polymerase chain
reaction fingerprint method. Typing of 2731 root nodule isolates revealed a total
of 38 fingerprint groups. More than 80% of the isolates could be grouped into six
dominant fingerprint groups, indicating that a few dominant bacterial strain
types had outcompeted the released strains.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16924432 [PubMed - indexed for MEDLINE]

1145: Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):12987-92. Epub 2006 Aug 21.

Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought
resistance and salt tolerance in rice.

Hu H, Dai M, Yao J, Xiao B, Li X, Zhang Q, Xiong L.

National Key Laboratory of Crop Genetic Improvement, National Center of Plant
Gene Research, Wuhan 430070, China.

Drought and salinity are major abiotic stresses to crop production. Here, we show
that overexpression of stress responsive gene SNAC1 (STRESS-RESPONSIVE NAC 1)
significantly enhances drought resistance in transgenic rice (22-34% higher seed
setting than control) in the field under severe drought stress conditions at the
reproductive stage while showing no phenotypic changes or yield penalty. The
transgenic rice also shows significantly improved drought resistance and salt
tolerance at the vegetative stage. Compared with WT, the transgenic rice are more
sensitive to abscisic acid and lose water more slowly by closing more stomatal
pores, yet display no significant difference in the rate of photosynthesis. SNAC1
is induced predominantly in guard cells by drought and encodes a NAM, ATAF, and
CUC (NAC) transcription factor with transactivation activity. DNA chip analysis
revealed that a large number of stress-related genes were up-regulated in the
SNAC1-overexpressing rice plants. Our data suggest that SNAC1 holds promising
utility in improving drought and salinity tolerance in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16924117 [PubMed - indexed for MEDLINE]

1146: Crit Rev Biotechnol. 2006 Jul-Sep;26(3):121-43.

Biotechnological methods to accelerate cheddar cheese ripening.

Azarnia S, Robert N, Lee B.

Department of Food Science and Agricultural Chemistry, McGill University,
Ste-Anne-de-Bellevue, QC, Canada.

Cheese is one of the dairy products that can result from the enzymatic
coagulation of milk. The basic steps of the transformation of milk into cheese
are coagulation, draining, and ripening. Ripening is the complex process required
for the development of a cheese's flavor, texture and aroma. Proteolysis,
lipolysis and glycolysis are the three main biochemical reactions that are
responsible for the basic changes during the maturation period. As ripening is a
relatively expensive process for the cheese industry, reducing maturation time
without destroying the quality of the ripened cheese has economic and
technological benefits. Elevated ripening temperatures, addition of enzymes,
addition of cheese slurry, attenuated starters, adjunct cultures, genetically
engineered starters and recombinant enzymes and microencapsulation of ripening
enzymes are traditional and modern methods used to accelerate cheese ripening. In
this context, an up to date review of Cheddar cheese ripening is presented.

Publication Types:
Review

PMID: 16923531 [PubMed - indexed for MEDLINE]

1147: Public Health Nutr. 2006 Aug;9(5):662-3.

Comment on:
Public Health Nutr. 2005 Sep;8(6A):673-94.

How far should nutrition reach?

Kent G.

Publication Types:
Comment
Letter

PMID: 16923303 [PubMed - indexed for MEDLINE]

1148: Biotechnol Appl Biochem. 2007 Jan;46(Pt 1):51-5.

Maize (Zea mays) genetic transformation by co-cultivating germinating seeds with
Agrobacterium tumefaciens.

Wang J, Sun Y, Li Y.

The Agri-Biotechnology Research Centre of Shanxi Province, 030031 Taiyuan,
People's Republic of China.

A novel plant genetic transformation method for maize (Zea mays) is reported.
Using a scalpel, germinating seeds were wounded in the meristem and co-cultivated
with an Agrobacterium tumefaciens strain harbouring a Ti plasmid. Seedlings
produced from the treatment were screened by hygromycin selection. Fertile
transgenic T(0) and T(1) plants were obtained. PCR amplification, PCR-Southern
and Southern-blot analysis showed that the foreign gene had been introduced into
the inbreds of maize. About 29% of T(0) seedlings examined were found to be
transgenic, although the overall transformation was only 0.6% when total treated
seeds were taken into account. The method circumvented tedious and prolonged
tissue-culture steps, is simple and can be readily integrated into conventional
plant breeding programs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16922678 [PubMed - indexed for MEDLINE]

1149: Plant Physiol. 2006 Oct;142(2):710-21. Epub 2006 Aug 18.

Suppression of LX ribonuclease in tomato results in a delay of leaf senescence
and abscission.

Lers A, Sonego L, Green PJ, Burd S.

Department of Postharvest Science of Fresh Produce, Agricultural Research
Organization, Volcani Center, Bet Dagan 50250, Israel. alers@volcani.agri.gov.il

Although present in different organisms and conserved in their protein sequence,
the biological functions of T2 ribonucleases (RNase) are generally unknown.
Tomato (Lycopersicon esculentum) LX is a T2/S-like RNase and its expression is
known to be associated with phosphate starvation, ethylene responses, and
senescence and programmed cell death. In this study, LX function was investigated
using antisense tomato plants in which the LX protein level was reduced. LX
protein levels normally become elevated when leaves senesce and antisense
inhibition of LX retarded the progression of senescence. Moreover, we observed a
marked delay of leaf abscission in LX-deficient plants. This correlated with
specific induction of LX protein in the tomato mature abscission zone tissue. LX
RNase gene regulation and the consequences of antisense inhibition indicate that
LX has an important functional role in both abscission and senescence.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16920876 [PubMed - indexed for MEDLINE]

1150: Plant Physiol. 2006 Oct;142(2):429-40. Epub 2006 Aug 18.

Computational estimation and experimental verification of off-target silencing
during posttranscriptional gene silencing in plants.

Xu P, Zhang Y, Kang L, Roossinck MJ, Mysore KS.

Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, Oklahoma
73401, USA.

Successful application of posttranscriptional gene silencing (PTGS) for gene
function study in both plants and animals depends on high target specificity and
silencing efficiency. By computational analysis with genome and/or transcriptome
sequences of 25 plant species, we predicted that about 50% to 70% of gene
transcripts in plants have potential off-targets when used for PTGS that could
obscure experimental results. We have developed a publicly available Web-based
computational tool called siRNA Scan to identify potential off-targets during
PTGS. Some of the potential off-targets obtained from this tool were tested by
measuring the amount of off-target transcripts using quantitative reverse
transcription-PCR. Up to 50% of the predicted off-target genes tested in plants
were actually silenced when tested experimentally. Our results suggest that a
high risk of off-target gene silencing exists during PTGS in plants. Our siRNA
Scan tool is useful to design better constructs for PTGS by minimizing off-target
gene silencing in both plants and animals.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16920874 [PubMed - indexed for MEDLINE]

1151: J Nutr. 2006 Sep;136(9):2331-7.

Transgenic flavonoid tomato intake reduces C-reactive protein in human C-reactive
protein transgenic mice more than wild-type tomato.

Rein D, Schijlen E, Kooistra T, Herbers K, Verschuren L, Hall R, Sonnewald U,
Bovy A, Kleemann R.

Friedrich-Alexander-Universitö¤t Erlangen-Nö¼rnberg, D-91058 Erlangen, Germany.

The increased consumption of fruits and vegetables is associated with reduced
cardiovascular disease. The molecular basis of this health effect is not fully
understood, yet dietary flavonoids are thought to play an important role. Genetic
engineering has enabled us to overexpress specific flavonoids (flavones and
flavonols) in tomato fruit. Human C-reactive protein transgenic (CRPtg) mice
express markers of cardiovascular risk that allow us to study of the putative
health effects of wild-type tomato (wtTom) and flavonoid-enriched tomato (flTom).
In this study, we analyzed whether consumption of wtTom, at a dose achievable
with a human diet, has beneficial effects on cardiovascular risk markers and
whether flTom may enhance such effects. CRPtg mice were fed a diet containing 4
g/kg wtTom, flTom peel, vehicle, or 1 g/kg fenofibrate, which reportedly reduces
cardiovascular risk, for 7 wk. Markers of general health (bodyweight, food
intake, and plasma alanine aminotransferase activities) and of cardiovascular
risk (plasma CRP, fibrinogen, E-selectin, and cholesterol levels) were analyzed.
All groups had comparable food intakes and body-weight gains. Plasma alanine
aminotransferase activities increased significantly in vehicle and
fenofibrate-treated mice. Compared with baseline, wtTom and flTom significantly
reduced basal human CRP concentrations by 43 and 56%, respectively. The
CRP-lowering effect of flTom significantly exceeded that of wtTom. The effects of
flTom on CRP were reversed within a 2-wk washout period. WtTom and flTom did not
affect fibrinogen, but comparably repressed E-selectin expression and upregulated
HDL cholesterol. Tomato peel consumption improved cardiovascular risk factors in
CRPtg mice, a beneficial effect that was further enhanced by enrichment of the
flavonoid content.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16920850 [PubMed - indexed for MEDLINE]

1152: Plant Cell. 2006 Sep;18(9):2294-313. Epub 2006 Aug 18.

Tung tree DGAT1 and DGAT2 have nonredundant functions in triacylglycerol
biosynthesis and are localized to different subdomains of the endoplasmic
reticulum.

Shockey JM, Gidda SK, Chapital DC, Kuan JC, Dhanoa PK, Bland JM, Rothstein SJ,
Mullen RT, Dyer JM.

U.S. Department of Agriculture, Agricultural Research Service, Southern Regional
Research Center, New Orleans, Louisiana 70124, USA.

Seeds of the tung tree (Vernicia fordii) produce large quantities of
triacylglycerols (TAGs) containing approximately 80% eleostearic acid, an unusual
conjugated fatty acid. We present a comparative analysis of the genetic,
functional, and cellular properties of tung type 1 and type 2 diacylglycerol
acyltransferases (DGAT1 and DGAT2), two unrelated enzymes that catalyze the
committed step in TAG biosynthesis. We show that both enzymes are encoded by
single genes and that DGAT1 is expressed at similar levels in various organs,
whereas DGAT2 is strongly induced in developing seeds at the onset of oil
biosynthesis. Expression of DGAT1 and DGAT2 in yeast produced different types and
proportions of TAGs containing eleostearic acid, with DGAT2 possessing an
enhanced propensity for the synthesis of trieleostearin, the main component of
tung oil. Both DGAT1 and DGAT2 are located in distinct, dynamic regions of the
endoplasmic reticulum (ER), and surprisingly, these regions do not overlap.
Furthermore, although both DGAT1 and DGAT2 contain a similar C-terminal
pentapeptide ER retrieval motif, this motif alone is not sufficient for their
localization to specific regions of the ER. These data suggest that DGAT1 and
DGAT2 have nonredundant functions in plants and that the production of storage
oils, including those containing unusual fatty acids, occurs in distinct ER
subdomains.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16920778 [PubMed - indexed for MEDLINE]

1153: Food Chem Toxicol. 2006 Nov;44(11):1909-15. Epub 2006 Jul 4.

Investigation on possible allergenicity of 19 different commercial enzymes used
in the food industry.

Bindslev-Jensen C, Skov PS, Roggen EL, Hvass P, Brinch DS.

Department of Dermatology and Allergy Center, Odense University Hospital, DK 5000
Odense C, Denmark. Carsten.Bindslev-Jensen@ouh.fyns-amt.dk

The aim of the study was to investigate the safety to allergic patients of 19
commercially available and authority-approved enzymes used in the food industry.
Enzymes produced by genetically modified organisms were included. Four hundred
consecutive adult patients with a diagnosed allergy to inhalation allergens, food
allergens, bee or wasp were included. All had at least one positive skin prick
test to the above allergens. Skin prick testing with the 19 enzymes was performed
on the forearm and if positive (in 13 patients), in vitro histamine release from
blood basophils were performed. Patients with positive results in skin prick test
were subsequently reinvestigated with further purified enzymes and finally
challenged orally with the enzymes in a double-blind, placebo-controlled
protocol. Only one reaction to a placebo challenge was seen. In some instances a
positive skin prick test result or a positive histamine release was seen elicited
by the enzymes, but since none of the patients were positive to any of the
commercial enzymes in the subsequent oral challenges using exaggerated dosages of
the enzymes compared to normal daily intake, the findings are without clinical
relevance. A wide variety of enzyme classes and origins was included in the
study. Because there were no allergenic findings of clinical relevance it is
concluded that ingestion of food enzymes in general is not considered to be a
concern with regard to food allergy.

Publication Types:
Clinical Trial
Controlled Clinical Trial

PMID: 16920243 [PubMed - indexed for MEDLINE]

1154: Phytochemistry. 2006 Oct;67(20):2215-24. Epub 2006 Aug 22.

Molecular cloning and heterologous expression of beta1,2-xylosyltransferase and
core alpha1,3-fucosyltransferase from maize.

Bondili JS, Castilho A, Mach L, Glö¶ssl J, Steinkellner H, Altmann F, Strasser R.

Institute of Applied Genetics and Cell Biology, University of Natural Resources
and Applied Life Sciences, BOKU-Vienna, Muthgasse 18, A-1190 Vienna, Austria.

Maize is considered a promising alternative production system for
pharmaceutically relevant proteins. However, like in all other plant species
asparagine-linked oligosaccharides of maize glycoproteins are modified with
beta1,2-xylose and core alpha1,3-fucose sugar residues, which are considered to
be immunogenic in mammals. This altered N-glycosylation when compared to
mammalian cells may reduce the potential of maize as a production system for
heterologous glycoproteins. Here we report the cloning and characterization of
the cDNA sequences coding for the maize enzymes beta1,2-xylosyltransferase (XylT)
and core alpha1,3-fucosyltransferase (FucT). The cloned XylT and FucT cDNAs were
shown to encode enzymatically active proteins, which were independently able to
convert a mammalian acceptor glycoprotein into an antigen binding anti-plant
N-glycan antibodies. The complete sequence of the XylT gene was determined.
Evidence for the presence of at least three XylT and FucT gene loci in the maize
genome was obtained. The identification of the two enzymes and their genes will
allow the targeted downregulation or even elimination of beta1,2-xylose and core
alpha1,3-fucose addition to recombinant glycoproteins produced in maize.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16920165 [PubMed - indexed for MEDLINE]

1155: J Plant Physiol. 2007 Aug;164(8):969-79. Epub 2006 Aug 21.

OsWRKY71, a rice transcription factor, is involved in rice defense response.

Liu X, Bai X, Wang X, Chu C.

National Key Laboratory of Plant Genomics and Plant Gene Research Center
(Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of
Sciences, Datun Road, Beijing 100101, China.

WRKY proteins are a large family of transcription factors that mainly participate
in plant biotic stress responses. So far, one hundred and five OsWRKY genes have
been predicted in the rice genome. To identify OsWRKY genes that might function
in inducible defense responses, a phylogenetic tree including 184 WRKY proteins
from Arabidopsis thaliana, rice, and other species was constructed. Based on the
phylogenetic analysis, ten candidate OsWRKY genes that may be involved in defense
responses were isolated from salicylic acid (SA)-treated rice seedlings. One of
them, OsWRKY71, was up-regulated by several defense signaling molecules, such as
SA, methyl jasmonate (MeJA), 1-aminocyclo-propane-1-carboxylic acid (ACC), as
well as wounding and pathogen infection, suggesting that OsWRKY71 might function
in rice biotic stress response. Transient expression of OsWRKY71:GFP fusion
protein in onion epidermis cells revealed that OsWRKY71 was localized in the
nucleus. Overexpression of OsWRKY71 gene in rice resulted in enhanced resistance
to virulent bacterial pathogens Xanthomonas oryzae pv. oryzae (Xoo) 13751.
Furthermore, two marker genes in defense signaling pathway, OsNPR1 and OsPR1b,
were constitutively expressed in OsWRKY71-overexpressing transgenic plants. These
results suggest that OsWRKY71 might function as a transcriptional regulator
upstream of OsNPR1 and OsPR1b in rice defense signaling pathways.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16919842 [PubMed - indexed for MEDLINE]

1156: Protein Expr Purif. 2007 Jan;51(1):22-7. Epub 2006 Jul 15.

Synthesis and assembly of Escherichia coli heat-labile enterotoxin B subunit in
transgenic lettuce (Lactuca sativa).

Kim TG, Kim MY, Kim BG, Kang TJ, Kim YS, Jang YS, Arntzen CJ, Yang MS.

Division of Biological Sciences and Research Center for Bioactive Materials,
Chonbuk National University, Jeonju 561-756, Republic of Korea.

Escherichia coli heat-labile enterotoxin B subunit (LTB) strongly induces immune
responses and can be used as an adjuvant for co-administered antigens. Synthetic
LTB (sLTB) based on optimal codon usage by plants was introduced into lettuce
cells (Lactuca sativa) by Agrobacterium tumefaciens-mediated transformation
methods. The sLTB gene was detected in the genomic DNA of transgenic lettuce leaf
cells by PCR DNA amplification. Synthesis and assembly of the sLTB protein into
oligomeric structures of pentameric size was observed in transgenic plant
extracts using Western blot analysis. The binding of sLTB pentamers to intestinal
epithelial cell membrane glycolipid receptors was confirmed by G(M1)-ganglioside
enzyme-linked immunosorbent assay (G(M1)-ELISA). Based on the results of ELISA,
sLTB protein comprised approximately 1.0-2.0% of total soluble protein in
transgenic lettuce leaf tissues. The synthesis and assembly of sLTB monomers into
biologically active oligomers in transgenic lettuce leaf tissues demonstrates the
feasibility of the use of edible plant-based vaccines consumed in the form of raw
plant materials to induce mucosal immunity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16919472 [PubMed - indexed for MEDLINE]

1157: J AOAC Int. 2006 Jul-Aug;89(4):913-28.

Immunoassay as an analytical tool in agricultural biotechnology.

Grothaus GD, Bandla M, Currier T, Giroux R, Jenkins GR, Lipp M, Shan G, Stave JW,
Pantella V.

EnviroLogix Inc, 500 Riverside Industrial Pkwy, Portland, ME 04103, USA.
davidgrothaus@envirologix.com

Immunoassays for biotechnology engineered proteins are used by AgBiotech
companies at numerous points in product development and by feed and food
suppliers for compliance and contractual purposes. Although AgBiotech companies
use the technology during product development and seed production, other
stakeholders from the food and feed supply chains, such as commodity, food, and
feed companies, as well as third-party diagnostic testing companies, also rely on
immunoassays for a number of purposes. The primary use of immunoassays is to
verify the presence or absence of genetically modified (GM) material in a product
or to quantify the amount of GM material present in a product. This article
describes the fundamental elements of GM analysis using immunoassays and
especially its application to the testing of grains. The 2 most commonly used
formats are lateral flow devices (LFD) and plate-based enzyme-linked
immunosorbent assays (ELISA). The main applications of both formats are discussed
in general, and the benefits and drawbacks are discussed in detail. The document
highlights the many areas to which attention must be paid in order to produce
reliable test results. These include sample preparation, method validation,
choice of appropriate reference materials, and biological and instrumental
sources of error. The article also discusses issues related to the analysis of
different matrixes and the effects they may have on the accuracy of the
immunoassays.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 16915826 [PubMed - indexed for MEDLINE]

1158: Plant Mol Biol. 2006 Sep;62(1-2):165-79. Epub 2006 Aug 17.

Subcellular pyrophosphate metabolism in developing tubers of potato (Solanum
tuberosum).

Farrö© EM, Tech S, Trethewey RN, Fernie AR, Willmitzer L.

Max-Planck-Institut fö¼r Molekulare Pflanzenphysiologie, Am Mö¼hlenberg 1, 14476
Golm, Germany. efarre@scripps.edu

PPi has previously been implicated specifically in the co-ordination of the
sucrose-starch transition and in the broader context of its role as co-factor in
heterotrophic plant metabolism. In order to assess the compartmentation of
pyrophosphate (PPi) metabolism in the potato tuber we analysed the effect of
expressing a bacterial pyrophosphatase in the amyloplast of wild type tubers or
in the cytosol or amyloplast of invertase-expressing tubers. The second and third
approaches were adopted since we have previously characterized the invertase
expressing lines to both exhibit highly altered sucrose metabolism and to contain
elevated levels of PPi (Farrö© et al. (2000a) Plant Physiol 123:681) and therefore
this background rendered questions concerning the level of communication between
the plastidic and cytosolic pyrophosphate pools relatively facile. In this study
we observed that the increase in PPi in the invertase expressing lines was mainly
confined to the cytosol. Accordingly, the expression of a bacterial
pyrophosphatase in the plastid of either wild type or invertase-expressing tubers
did not lead to a decrease in total PPi content. However, the expression of the
heterologous pyrophosphatase in the cytosol of cytosolic invertase-expressing
tubers led to strong metabolic changes. These results are discussed both with
respect to our previous hypotheses and to current models of the compartmentation
of potato tuber metabolism.

PMID: 16915524 [PubMed - indexed for MEDLINE]

1159: Plant Mol Biol. 2006 Sep;62(1-2):195-214. Epub 2006 Aug 17.

A novel cis-acting element, ESP, contributes to high-level endosperm-specific
expression in an oat globulin promoter.

Vickers CE, Xue G, Gresshoff PM.

CSIRO Plant Industry, 306 Carmody Rd, St Lucia, Brisbane 4067, Australia.
cvickers@essex.ac.uk

To examine the genetic controls of endosperm (ES) specificity, several cereal
seed storage protein (SSP) promoters were isolated and studied using a transient
expression analysis system. An oat globulin promoter (AsGlo1) capable of driving
strong ES-specific expression in barley and wheat was identified. Progressive 5'
deletions and cis element mutations demonstrated that the mechanism of
specificity in the AsGlo1 promoter was distinct from that observed in glutelin
and prolamin promoters. A novel interrupted palindromic sequence, ACATGTCATCATGT,
was required for ES specificity and substantially contributed to expression
strength of the AsGlo1 promoter. This sequence was termed the endosperm
specificity palindrome (ESP) element. The GCN4 element, which has previously been
shown to be required for ES specificity in cereal SSP promoters, had a
quantitative role but was not required for tissue specificity. The 960-bp AsGlo1
promoter and a 251-bp deletion containing the ESP element also drove ES-specific
expression in stably transformed barley. Reporter gene protein accumulated at
very high levels (10% of total soluble protein) in ES tissues of plants
transformed with an AsGlo1:GFP construct. Expression strength and tissue
specificity were maintained over five transgenic generations. These attributes
make the AsGlo1 promoter an ideal promoter for biotechnology applications. In
conjunction with previous findings, our data demonstrate that there is more than
one genetically distinct mechanism by which ES specificity can be achieved in
cereal SSP promoters, and also suggest that there is redundancy between
transcriptional and post-transcriptional tissue specificity mechanisms in cereal
globulin genes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16915522 [PubMed - indexed for MEDLINE]

1160: Plant Mol Biol. 2006 Oct;62(3):439-52. Epub 2006 Aug 17.

The rice OsGAE1 is a novel gibberellin-regulated gene and involved in rice
growth.

Jan A, Kitano H, Matsumoto H, Komatsu S.

Department of Molecular Biology, National Institute of Agrobiological Sciences,
2-1-2 Kannondai, Tsukuba 305-8602, Japan.

Gibberellins (GAs) are a class of phytohormones that regulate many aspects of
plant growth and development processes including stem elongation, flowering, and
seed germination. A novel GA-enhanced gene, designated as OsGAE1, was identified
using microarray analysis of GA-regulated genes. OsGAE1 expressed in a dose- and
time course-dependent manner with minimum expression at 1 microM GA(3) and
maximum expression at 50 microM GA(3) starting from 30 min and peaked at 24 h
after GA(3) treatment. OsGAE1 expression was up-regulated by GA(3) at transcript
level while no significant effect was observed for other hormones. OsGAE1 was
expressed in Escherichia coli with N-terminal His(6) tag and the recombinant
protein migrated at 38 kDa, slightly larger than the predicted 29 kDa, during
SDS-PAGE. Anti-OsGAE1 antibodies immunoreacted with a protein of 40 kDa in rice
leaf sheath. OsGAE1 expressed mainly in growing leaf sheath and callus compared
to leaf and root. In situ hybridization and OsGAE1 promoter analysis revealed
that OsGAE1 expressed in shoot apex meristem and young primary leaves. Northern
blot, Western blot, and GUS activities revealed that OsGAE1 is up-regulated by
GA(3). Transgenic rice expressing OsGAE1 in antisense orientation exhibited
severely affected vegetative and reproductive growth. The transgenic plants were
55-70% short compared to control. These results suggest that OsGAE1 is
differentially expressed in rice leaf sheath in relation to GA(3) and it encodes
a functional protein which is involved in GA-regulated growth and development of
rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16915516 [PubMed - indexed for MEDLINE]

1161: Plant Cell Environ. 2006 Sep;29(9):1829-40.

Are ABA, ethylene or their interaction involved in the response of leaf growth to
soil water deficit? An analysis using naturally occurring variation or genetic
transformation of ABA production in maize.

Voisin AS, Reidy B, Parent B, Rolland G, Redondo E, Gerentes D, Tardieu F, Muller
B.

LEPSE, INRA, 2 place Viala, 34060 Montpellier, France.

The role of abscisic acid (ABA) and its possible interaction with ethylene in
mediating leaf elongation response to soil water deficit are a matter of
controversy. To address this question, we used a set of maize genotypes with
various levels of ABA either due to natural variability or to genetic
transformation targeted on NCED/VP14, a key enzyme of ABA synthesis. The
transgenic lines yielded less strong phenotypes than available mutants, making it
possible to use them under normal growing conditions. We focused on leaf
elongation during night periods in order to avoid the confounding effect of ABA
on leaf water status. Our results suggest that over a wide range, internal ABA
level (measured in both leaf extracts or xylem sap) has no clear effect on leaf
elongation response to soil water deficit, except in the case of an antisense
line presenting the strongest reduction in ABA accumulation that showed a slight
maintenance of leaf elongation during water deficit. Leaf ethylene production
rate was variable and not related to water deficit except in the ABA-deficient
transgenic lines where it was increased by water deficit on average but not
systematically. Moreover, variability in ethylene production rate was not linked
to variability in elongation rate. Our results thus suggest that neither ABA nor
ethylene seems to play a major role in the control of leaf elongation response to
soil water deficit.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16913872 [PubMed - indexed for MEDLINE]

1162: Biotechnol Lett. 2006 Oct;28(20):1661-6. Epub 2006 Aug 16.

Expression of Helicobacter pylori urease subunit B gene in transgenic rice.

Gu Q, Han N, Liu J, Zhu M.

Department of Biotechnology, Zhejiang Gongshang University, Hangzhou, 310035,
China. guqing2002@hotmail.com

Helicobacter pylori ureB antigen gene was cloned to the 5'-end of gus
(beta-glucuronidase) reporter gene between CaMV35S promoter and the octopine
synthase (OCS) terminator in the plasmid, pCAMBIA13011. It was then introduced
into rice genome by Agrobacterium-mediated transformation. A total of 30
regenerated plants with hygromycin resistance were obtained in the selection
media. The putative transgenic individuals were tested for the presence of ureB
in the nuclear genome of rice plants by PCR analysis. Expression of ureB gene in
rice plants was verified by RT-PCR and Western blot analysis using polyclonal
human antiserum for transcription and translation levels respectively. These
results provide a basis for further studies on the accumulation level of UreB
recombinant protein in transgenic rice and potential utilization of transgenic
rice for delivery of edible vaccines against Helicobacter pylori.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16912927 [PubMed - indexed for MEDLINE]

1163: Plant Mol Biol. 2006 Sep;62(1-2):71-82. Epub 2006 Aug 16.

Self-excision of the antibiotic resistance gene nptII using a heat inducible
Cre-loxP system from transgenic potato.

Cuellar W, Gaudin A, Solö¨rzano D, Casas A, Nopo L, Chudalayandi P, Medrano G,
Kreuze J, Ghislain M.

Applied Biotechnology Laboratory, Germplasm enhancement and Crop Improvement
Division, International Potato Center CIP, P.O. Box 1558, Lima 12, Peru.

Resistance to antibiotics mediated by selectable marker genes remains a powerful
selection tool for transgenic event production. However, regulatory agencies and
consumer concerns favor these to be eliminated from food crops. Several excision
systems exist but none have been optimized or shown to be functional for clonally
propagated crops. The excision of the nptII gene conferring resistance to
kanamycin has been achieved here using a gene construct based on a heat-inducible
cre gene producing a recombinase that eliminates cre and nptII genes flanked by
two loxP sites. First-generation regenerants with the Cre-loxP system were
obtained by selection on kanamycin media. Following a heat treatment, second
generation regenerants were screened for excision by PCR using nptII, cre, and
T-DNA borders primers. Excision efficiency appeared to be at 4.7% depending on
the heat treatment. The footprint of the excision was shown by sequencing between
T-DNA borders to correspond to a perfect recombination event. Selectable
marker-free sprouts were also obtained from tubers of transgenic events when
submitted to similar heat treatment at 4% frequency. Spontaneous excision was not
observed out of 196 regenerants from untreated transgenic explants. Biosafety
concerns are minimized because the expression of cre gene driven by the hsp70
promoter of Drosophila melanogaster was remarkably low even under heat activation
and no functional loxP site were found in published Solanum sequence database. A
new plant transformation vector pCIP54/55 was developed including a multiple
cloning site and the self-excision system which should be a useful tool not only
for marker genes in potato but for any gene or sequence removal in any plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16912912 [PubMed - indexed for MEDLINE]

1164: Plant Mol Biol. 2006 Oct;62(3):385-95. Epub 2006 Aug 16.

The involvement of chloroplast HSP100/ClpB in the acquired thermotolerance in
tomato.

Yang JY, Sun Y, Sun AQ, Yi SY, Qin J, Li MH, Liu J.

College of Life Science, Shandong Normal University, Jinan, Shandong, 250014, PR
China.

The chloroplast HSP100/ClpB is a newly documented member of the ClpB family, but
little was known about its role in imparting thermotolerance to cells. A cDNA
coding for a HSP100/ClpB homolog has been cloned from Lycopersicon esculentum and
termed as Lehsp100/ClpB (the cDNA sequence of Lehsp100/ClpB has been submitted to
the GenBank database under accession number: AB219939). The protein encoded by
the cDNA was most similar to the putative chloroplast HSP100/ClpBs in higher
plants and the ClpB from Cyanobacterium Synechococcus sp. A 97 kDa protein, which
matched the predicted size of mature LeHSP100/ClpB, was immunologically detected
in chloroplast isolated from heat-treated tomato plants. In addition, the fusion
protein, combining the transit sequence of LeHSP100/ClpB and GFP, was found to be
located in chloroplast based on the observations of fluorescent microscope
images. These results indicated the chloroplast-localization of LeHSP100/ClpB.
Both the transcript and the protein of Lehsp100/ClpB were not detected under
normal growth conditions, but they were induced by increasingly higher
temperatures. An antisense Lehsp100/ClpB cDNA fragment was introduced into the
tomato by Agrobacterium-mediated transformation. Antisense lines exhibited an
extreme repression of heat-induced expression of Lehsp100/ClpB. The levels of
chloroplast HSP60 and small HSP in antisense lines were identical to those of the
control plants. After plants preconditioned at 38 degrees C for 2 h were exposed
to a lethal heat shock at 46 degrees C for 2 h, the antisense lines were greatly
impaired and withered in 21 days of the recovery phase, whereas the untransformed
control plants and the vector-transformed plants survived. Furthermore,
chlorophyll fluorescence measurements showed that PS II in antisense lines were
more susceptible to the thermal irreversible inactivation than the untransformed
and vector-transformed control plants. This work provides the first example that
induction of chloroplast LeHSP100/ClpB contributes to the acquisition of
thermotolerance in higher plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16912911 [PubMed - indexed for MEDLINE]

1165: J Virol. 2006 Sep;80(17):8459-68.

Membrane-bound tomato mosaic virus replication proteins participate in RNA
synthesis and are associated with host proteins in a pattern distinct from those
that are not membrane bound.

Nishikiori M, Dohi K, Mori M, Meshi T, Naito S, Ishikawa M.

Plant-Microbe Interactions Research Unit, Division of Plant Sciences, National
Institute of Agrobiological Sciences, Tsukuba, Japan.

Extracts of vacuole-depleted, tomato mosaic virus (ToMV)-infected plant
protoplasts contained an RNA-dependent RNA polymerase (RdRp) that utilized an
endogenous template to synthesize ToMV-related positive-strand RNAs in a pattern
similar to that observed in vivo. Despite the fact that only minor fractions of
the ToMV 130- and 180-kDa replication proteins were associated with membranes,
the RdRp activity was exclusively associated with membranes. A genome-sized,
negative-strand RNA template was associated with membranes and was resistant to
micrococcal nuclease unless treated with detergents. Non-membrane-bound
replication proteins did not exhibit RdRp activity, even in the presence of ToMV
RNA. While the non-membrane-bound replication proteins remained soluble after
treatment with Triton X-100, the same treatment made the membrane-bound
replication proteins in a form that precipitated upon low-speed centrifugation.
On the other hand, the detergent lysophosphatidylcholine (LPC) efficiently
solubilized the membrane-bound replication proteins. Upon LPC treatment, the
endogenous template-dependent RdRp activity was reduced and exogenous ToMV RNA
template-dependent RdRp activity appeared instead. This activity, as well as the
viral 130-kDa protein and the host proteins Hsp70, eukaryotic translation
elongation factor 1A (eEF1A), TOM1, and TOM2A copurified with FLAG-tagged viral
180-kDa protein from LPC-solubilized membranes. In contrast, Hsp70 and only small
amounts of the 130-kDa protein and eEF1A copurified with FLAG-tagged
non-membrane-bound 180-kDa protein. These results suggest that the viral
replication proteins are associated with the intracellular membranes harboring
TOM1 and TOM2A and that this association is important for RdRp activity.
Self-association of the viral replication proteins and their association with
other host proteins may also be important for RdRp activity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16912296 [PubMed - indexed for MEDLINE]

1166: Eur Rev Med Pharmacol Sci. 2006 Jul-Aug;10(4):197-206.

Benefits and concerns associated with biotechnology-derived foods: can additional
research reduce children health risks?

Cantani A.

Allergy and Clinical Immunology Division, Pediatric Department, La Sapienza
University, Rome, Italy.

The development of techniques devised for the genetic manipulation of foods poses
new risks for children with food allergy (FA). The introduction of foreign
allergenic proteins from different foods into previously tolerated foods may
trigger allergic reactions, often complicating with anaphylactic shock in a
subset of allergic babies. Children with FA, even if subjected to preventative
diets, always challenge the risk of developing allergic manifestations after
unintentional intake of a non tolerated food in restaurant settings, with
relatives or schoolmates, etc, where product labelling is necessarily lacking.
The introduction of potentially allergenic proteins into foods generally
considered safe for allergic children can be done deliberately, by either
substantially altering the food ingredients, or by genetic manipulation which
change the composition or transfer allergens, or unintentionally by
quality-control failures, due to contaminations in the production process, or to
genetic mismanipulation. There is a controversy between multinationals often
favored by governments and consumer association resistance, thus an equidistant
analysis poses some unprecedented impediments. The importance of FA and the
potential of transgenic plants to bring food allergens into the food supply
should not be disregarded. The expression in soybeans of a Brazil nut protein
resulted in a food allergen expressed in widely used infant formulas, so paving
the way to an often reported multinational debacle. Genetic engineering poses
innovative ethical and social concerns, as well as serious challenges to the
environment, human health, animal welfare, and the future of agriculture. In this
paper will be emphasized practical concepts more crucial for pediatricians.

Publication Types:
Review

PMID: 16910351 [PubMed - indexed for MEDLINE]

1167: J Environ Health. 2006 Jul-Aug;69(1):33-4.

The ethical dilemma of genetically modified food.

Jefferson V.

National Capital Area Environmental Health Association, Clinton, MD 20735, USA.
Val.Jefferson@verizon.net

PMID: 16910106 [PubMed - indexed for MEDLINE]

1168: Plant Cell Rep. 2007 Jan;26(1):61-70. Epub 2006 Aug 15.

Increased alpha-tocopherol content in soybean seed overexpressing the Perilla
frutescens gamma-tocopherol methyltransferase gene.

Tavva VS, Kim YH, Kagan IA, Dinkins RD, Kim KH, Collins GB.

Department of Plant and Soil Sciences, University of Kentucky, 1405 Veterans
Road, Lexington, KY 40546-0312, USA.

Tocopherols, with antioxidant properties, are synthesized by photosynthetic
organisms and play important roles in human and animal nutrition. In soybean,
gamma-tocopherol, the biosynthetic precursor to alpha-tocopherol, is the
predominant form found in the seed, whereas alpha-tocopherol is the most
bioactive component. This suggests that the final step of the alpha-tocopherol
biosynthetic pathway catalyzed by gamma-tocopherol methyltransferase (gamma-TMT)
is limiting in soybean seed. Soybean oil is the major edible vegetable oil
consumed, so manipulating the tocopherol biosynthetic pathway in soybean seed to
convert tocopherols into more active alpha-tocopherol form could have significant
health benefits. In order to increase the soybean seed alpha-tocopherol content,
the gamma-TMT gene isolated from Perilla frutescens was overexpressed in soybean
using a seed-specific promoter. One transgenic plant was recovered and the
progeny was analyzed for two generations. Our results demonstrated that the
seed-specific expression of the P. frutescens gamma-TMT gene resulted in a
10.4-fold increase in the alpha-tocopherol content and a 14.9-fold increase in
the beta-tocopherol content in T2 seed. Given the relative contributions of
different tocopherols to vitamin E activity, the activity in T2 seed was
calculated to be 4.8-fold higher than in wild-type seed. In addition, the data
obtained on lipid peroxidation indicates that alpha-tocopherol may have a role in
preventing oxidative damage to lipid components during seed storage and seed
germination. The increase in the alpha-tocopherol content in the soybean seed
could have a potential to significantly increase the dietary intake of vitamin E.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16909228 [PubMed - indexed for MEDLINE]

1169: J Anim Sci. 2006 Sep;84(9):2346-51.

Effects of suckling intensity on milk yield and piglet growth from
lactation-enhanced gilts.

Marshall KM, Hurley WL, Shanks RD, Wheeler MB.

University of Illinois, Department of Animal Sciences, Urbana, IL 6180, USA.

The effects of suckling intensity on milk yield and piglet growth were determined
when lactation capacity of the sow was enhanced through overexpression of a
mammary-specific transgene, bovine alpha-lactalbumin. Lactational response to
increased suckling stimulation was determined by fostering litters of the same
age (d 1) or 7 d older (d 7) than the day of lactation to sows nontransgenic
(control) or transgenic (TG) for bovine alpha-lactalbumin. Twenty first-parity
gilts were allocated to 4 treatments dependent on gilt genotype and age of litter
fostered (control d 1, control d 7, TG d 1, and TG d 7). Litters were
standardized to 10 piglets within 24 h postpartum, and nonbirth piglets were
fostered to gilts with an equal litter BW within age groups at 36 h postpartum.
Milk yield was determined by the weigh-suckle-weigh method on d 6, 9, 12, 15, and
18 of lactation. Mean daily milk yield was greater (P = 0.031) for TG gilts
compared with control gilts and tended to be greater (P = 0.056) for all gilts
with d-7 piglets compared with those with d-1 piglets. Daily milk yield of TG d 7
gilts increased rapidly to peak at d 9 and was greater than milk yield of all
control gilts at d 9 (P < 0.01), 12 (P < 0.02), and 15 (P < 0.02). Mean daily
milk yield of TG d 7 gilts was 2.1 kg greater (P = 0.002) than for control d 7
gilts and 2.0 kg greater (P = 0.004) than for TG d 1 gilts. Daily milk yield of
control d 1 gilts was not different from that of TG d 1 gilts (P = 0.49) or
control d 7 gilts (P = 0.63). Piglet BW gain between d 3 and 6 was greater (P <
0.01) in the TG d 7 group than for all other groups and was greater (P < 0.05)
than the control groups between d 6 and 9. No difference was found when comparing
accumulated BW gain of the piglets between the day of age at foster (d 1 vs. 7; P
= 0.606) or between the control d 1 and control d 7 groups (P = 0.759).
Accumulated BW gain of piglets suckling TG d 7 gilts from d 3 through 9 was
greater (P < 0.02) than that of the other groups and continued to be greater (P <
0.05) than that of either of the control groups through d 15. However, by d 15,
accumulated BW gain of piglets suckling TG d 1 gilts was no longer different (P =
0.40) from that of the TG d 7 group and was greater (P < 0.05) than that of the
control d 1 group. The enhanced lactation potential of these TG gilts synergized
with suckling intensity to stimulate increased milk production during early
lactation, resulting in increased piglet growth.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16908636 [PubMed - indexed for MEDLINE]

1170: J Exp Bot. 2006;57(12):3069-78. Epub 2006 Aug 14.

High-level tryptophan accumulation in seeds of transgenic rice and its limited
effects on agronomic traits and seed metabolite profile.

Wakasa K, Hasegawa H, Nemoto H, Matsuda F, Miyazawa H, Tozawa Y, Morino K,
Komatsu A, Yamada T, Terakawa T, Miyagawa H.

Department of Rice Breeding, National Institute of Crop Science, 2-1-18
Kannondai, Tsukuba, Ibaraki 305-8518, Japan. k3wakasa@nodai.ac.jp

Metabolic manipulation of plants to improve their nutritional quality is an
important goal of plant biotechnology. Expression in rice (Oryza sativa L.) of a
transgene (OASA1D) encoding a feedback-insensitive alpha subunit of rice
anthranilate synthase results in the accumulation of tryptophan (Trp) in calli
and leaves. It is shown here that the amount of free Trp in the seeds of such
plants is increased by about two orders of magnitude compared with that in the
seeds of wild-type plants. The total Trp content in the seeds of the transgenic
plants was also increased. Two homozygous lines, HW1 and HW5, of OASA1D
transgenic rice were generated for characterization of agronomic traits and
aromatic metabolite profiling of seeds. The marked overproduction of Trp was
stable in these lines under field conditions, although spikelet fertility and
yield, as well as seed germination ability, were reduced compared with the wild
type. These differences in agronomic traits were small, however, in HW5. In spite
of the high Trp content in the seeds of the HW lines, metabolic profiling
revealed no substantial changes in the amounts of other phenolic compounds. The
amount of indole acetic acid was increased about 2-fold in the seeds of the
transgenic lines. The establishment and characterization of these OASA1D
transgenic lines have thus demonstrated the feasibility of increasing the Trp
content in the seeds of rice (or of other crops) as a means of improving its
nutritional value for human consumption or animal feed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16908506 [PubMed - indexed for MEDLINE]

1171: BMC Biotechnol. 2006 Aug 14;6:37.

Critical points of DNA quantification by real-time PCR--effects of DNA extraction
method and sample matrix on quantification of genetically modified organisms.

Cankar K, Stebih D, Dreo T, Zel J, Gruden K.

Department of Plant Physiology and Biotechnology, National Institute of Biology,
Vecna pot 111, 1000 Ljubljana, Slovenia. katja.cankar@nib.si

BACKGROUND: Real-time PCR is the technique of choice for nucleic acid
quantification. In the field of detection of genetically modified organisms
(GMOs) quantification of biotech products may be required to fulfil legislative
requirements. However, successful quantification depends crucially on the quality
of the sample DNA analyzed. Methods for GMO detection are generally validated on
certified reference materials that are in the form of powdered grain material,
while detection in routine laboratories must be performed on a wide variety of
sample matrixes. Due to food processing, the DNA in sample matrixes can be
present in low amounts and also degraded. In addition, molecules of plant origin
or from other sources that affect PCR amplification of samples will influence the
reliability of the quantification. Further, the wide variety of sample matrixes
presents a challenge for detection laboratories. The extraction method must
ensure high yield and quality of the DNA obtained and must be carefully selected,
since even components of DNA extraction solutions can influence PCR reactions.
GMO quantification is based on a standard curve, therefore similarity of PCR
efficiency for the sample and standard reference material is a prerequisite for
exact quantification. Little information on the performance of real-time PCR on
samples of different matrixes is available. RESULTS: Five commonly used DNA
extraction techniques were compared and their suitability for quantitative
analysis was assessed. The effect of sample matrix on nucleic acid quantification
was assessed by comparing 4 maize and 4 soybean matrixes. In addition 205 maize
and soybean samples from routine analysis were analyzed for PCR efficiency to
assess variability of PCR performance within each sample matrix. Together with
the amount of DNA needed for reliable quantification, PCR efficiency is the
crucial parameter determining the reliability of quantitative results, therefore
it was chosen as the primary criterion by which to evaluate the quality and
performance on different matrixes and extraction techniques. The effect of PCR
efficiency on the resulting GMO content is demonstrated. CONCLUSION: The crucial
influence of extraction technique and sample matrix properties on the results of
GMO quantification is demonstrated. Appropriate extraction techniques for each
matrix need to be determined to achieve accurate DNA quantification.
Nevertheless, as it is shown that in the area of food and feed testing matrix
with certain specificities is impossible to define strict quality controls need
to be introduced to monitor PCR. The results of our study are also applicable to
other fields of quantitative testing by real-time PCR.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 16907967 [PubMed - indexed for MEDLINE]

1172: J Appl Microbiol. 2006 Sep;101(3):616-9.

Beyond the spore--past and future developments of Bacillus thuringiensis as a
biopesticide.

Crickmore N.

School of Life Sciences, University of Sussex, Falmer, Brighton, UK.
n.crickmore@sussex.ac.uk

Formulated and sporulated cultures of Bacillus thuringiensis (Bt) are widely used
as foliar sprays as part of integrated pest management strategies against insect
pests of agricultural crops. Although in several cases the presence of the spore
has been shown to improve the activity of the product, other Bt-based
insecticides have been developed in which the spore is absent. The most notable
of these are transgenic plants expressing just the insect toxin gene from the
bacterium. This paper will discuss these developments, and the advantages and
disadvantages of having the spore present.

Publication Types:
Review

PMID: 16907811 [PubMed - indexed for MEDLINE]

1173: Pest Manag Sci. 2006 Oct;62(10):999-1012.

Comparing the impact of conventional pesticide and use of a transgenic
pest-resistant crop on the beneficial carabid beetle Pterostichus melanarius.

Mulligan EA, Ferry N, Jouanin L, Walters KF, Port GR, Gatehouse AM.

Institute for Research on Environment and Sustainability, School of Biology,
University of Newcastle Upon-Tyne, Newcastle NE1 7RU, UK.

The potential impact of a chemical pesticide control method has been compared
with that of transgenic plants expressing a protease inhibitor conferring insect
resistance by utilising a tritrophic system comprising the crop plant Brassica
napus (L.) (Oilseed rape), the pest mollusc Deroceras reticulatum (Mö¼ller) and
the predatory carabid beetle Pterostichus melanarius (Illiger). Cypermethrin, as
the most widely used pesticide in UK oilseed rape (OSR) cultivation, was selected
as the conventional treatment. OSR expressing a cysteine protease inhibitor,
oryzacystatin-1 (OC-1), was the transgenic comparator. In feeding trials, D.
reticulatum showed no significant long-term effects on measured life history
parameters (survival, weight gain, food consumption) as a result of exposure to
either the cypermethrin or OC-1 treatment. However, D. reticulatum was able to
respond to the presence of the dietary inhibitor by producing two novel proteases
following exposure to OC-1-expressing OSR. Similarly, P. melanarius showed no
detectable alterations in mortality, weight gain or food consumption when feeding
on D. reticulatum previously fed either pesticide-contaminated or GM plant
material. Furthermore, as with the slug, a novel form of protease, approximately
M(r) 27 kDa, was induced in the carabid in response to feeding on slugs fed
OC-1-expressing OSR.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16906504 [PubMed - indexed for MEDLINE]

1174: Transgenic Res. 2006 Aug;15(4):515-9.

Consumption of milk from transgenic goats expressing human lysozyme in the
mammary gland results in the modulation of intestinal microflora.

Maga EA, Walker RL, Anderson GB, Murray JD.

Department of Animal Science, University of California, One Shields Avenue,
Davis, CA 95616, USA. eamaga@ucdavis.edu

Lysozyme is a key antimicrobial component of human milk that has several
health-promoting functions including the development of a healthy intestinal
tract. However, levels of lysozyme in the milk of dairy animals are negligible.
We have generated transgenic dairy goats that express human lysozyme (HLZ) in
their milk in an attempt to deliver the benefits of human milk in a continual
fashion. To test the feasibility of this transgenic approach to achieve a
biological impact at the level of the intestine, feeding trials were conducted in
two animal models. Pasteurized milk from HLZ transgenic animals was fed to both
kid goats (ruminant model) and young pigs (human model), and the numbers of total
coliforms and Escherichia coli present in the small intestine were determined.
Data from this proof-of-principle study demonstrate that milk from transgenic
animals was capable of modulating the bacterial population of the gut in both
animal models. Pigs that consumed pasteurized milk from HLZ transgenic goats had
fewer numbers of coliforms and E. coli in their intestine than did those
receiving milk from non-transgenic control animals. The opposite effect was seen
in goats. Milk from these transgenic animals not only represent one of the first
transgenic food products with the potential of benefiting human health, but are
also a unique model to study the development and role of intestinal microflora on
health, well-being and resistance to disease.

PMID: 16906451 [PubMed - indexed for MEDLINE]

1175: Transgenic Res. 2006 Aug;15(4):481-8.

Expression of the cry9Aa2 B.t. gene in tobacco chloroplasts confers resistance to
potato tuber moth.

Chakrabarti SK, Lutz KA, Lertwiriyawong B, Svab Z, Maliga P.

Central Potato Research Institute, Shimla, 171001 HP, India.

We report here the control of potato tuber moth (Phthorimaea operculella) by
incorporating a truncated Bacillus thuringiensis cry9Aa2 gene in the plastid
genome. Plasmids pSKC84 and pSKC85 are derivatives of a new polycistronic plastid
transformation vector, pPRV312L, that carries spectinomycin resistance (aadA) as
a selective marker and targets insertions in the trnI-trnA intergenic region. The
Cry9Aa2 N-terminal region (82.1 kDa; 734 amino acids) was expressed in a
cassette, which consists of 49 nucleotides of the cry9Aa2 leader and the
3'-untranslated region of the plastid rbcL gene (TrbcL), and relies on
readthrough transcription from the plastid rRNA operon. In a tobacco leaf
bioassay, expression of Cry9Aa2 conferred resistance to potato tuber moth. In
accordance, the Cry9Aa2 insecticidal protein accumulated to high levels,
approximately 10% of the total soluble cellular protein and approximately 20% in
the membrane fraction. However, high-level Cry9Aa2 expression significantly
delayed plant development. Thus, a practical system to control potato tuber moth
by Cry9Aa2 expression calls for down-regulation of its expression.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16906448 [PubMed - indexed for MEDLINE]

1176: Transgenic Res. 2006 Aug;15(4):465-80.

Erratum in:
Transgenic Res. 2007 Apr;16(2):253-9.

Characterization and multi-generational stability of the growth hormone transgene
(EO-1alpha) responsible for enhanced growth rates in Atlantic Salmon.

Yaskowiak ES, Shears MA, Agarwal-Mawal A, Fletcher GL.

Aqua Bounty Canada, Inc., P.O. Box 13422, A1B 4B7, St. John's, Newfoundland,
Canada.

Transgenic technologies provide a promising means by which desirable traits can
be introduced into cultured fish species within a single generation thus
accelerating the production of genetically superior broodstock for aquaculture.
However, before such fish are allowed to be marketed as food they must receive
government regulatory approval. Two pivotal regulatory requirements are: (1)
complete characterization of the genomically integrated transgene and, (2)
demonstration that the transgene remains stable over multiple generations. We
have generated a stable line of growth hormone (GH) transgenic Atlantic salmon
(Salmo salar) using an "all fish" gene construct (opAFP-GHc2) containing a growth
hormone cDNA from chinook salmon whose expression is regulated by the 5' promoter
and 3' termination regions derived from an ocean pout antifreeze protein (AFP)
gene. In this study we show that a reorganized form of the opAFP-GHc2 construct
(termed EO-1alpha) integrated as a single functional copy into a 35 bp repeat
region of the genomic DNA. PCR based mapping revealed that the linear sequence of
the EO-1alpha integrant was organized as follows: base pairs 1580-2193 of the
ocean pout promoter region followed by the intact chinook salmon GH cDNA, the
complete ocean pout antifreeze 3' region, and the first 1678 bp of the ocean pout
antifreeze 5' region. Sequence analysis of the EO-1alpha integrant and genomic
flanking regions in F2 and F4 generation salmon revealed that they were
identical. In addition, apart from the disruption at the integration sites, the
consensus sequences of the integrant in these two generations of salmon were
identical to the sequence of the opAFP-GHc2 construct. These results indicate
that the EO-1alpha transgene codes for the chinook salmon GH, and that the
transgene and the integration site have remained stable over multiple
generations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16906447 [PubMed - indexed for MEDLINE]

1177: Transgenic Res. 2006 Aug;15(4):455-63.

Expression of the Newcastle disease virus fusion protein in transgenic maize and
immunological studies.

Guerrero-Andrade O, Loza-Rubio E, Olivera-Flores T, Fehö©rvö¡ri-Bone T, Gö¨mez-Lim
MA.

Departamento de Ingenierö­a Genö©tica de Plantas, Cinvestav Campus Guanajuato, Km
9.6 Libramiento Norte Carretera Irapuato-Leö¨n, Apdo, Postal 629, Irapuato,
Guanajuato, Mö©xico 36500.

Transgenic plants have been employed successfully as a low-cost system for the
production of therapeutically valuable proteins, including antibodies, antigens
and hormones. Here, we report the expression of the fusion (F) gene of the
Newcastle disease virus (NDV) in transgenic maize plants. The expression of the
transgene, driven by the maize ubiquitin promoter, caused accumulation of the F
protein in maize kernels. The presence of the transgene was verified by Southern
and western blots. Feeding chickens with kernels containing the F protein induced
the production of antibodies, which conferred protection against a viral
challenge. This protection was comparable to that conferred by a commercial
vaccine. Possible uses of this plant-based F protein as a potential mucosal
vaccine are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16906446 [PubMed - indexed for MEDLINE]

1178: Transgenic Res. 2006 Aug;15(4):435-46.

Expression of the lipid transfer protein Ace-AMP1 in transgenic wheat enhances
antifungal activity and defense responses.

Roy-Barman S, Sautter C, Chattoo BB.

Department of Microbiology and Biotechnology Centre, Faculty of Science, The M.
S. University of Baroda, Vadodara 390 002, India.

To enhance fungal disease resistance, wheat plants (cv. Bobwhite) were engineered
to constitutively express the potent antimicrobial protein Ace-AMP1 from Allium
cepa, driven by a maize ubiquitin promoter along with its first intron. The bar
gene was used for selection of putative transformants on medium containing
phosphinothricin (PPT). Transgene inheritance, integration and stability of
expression were confirmed over two generations by PCR, Southern, northern and
western blot analyses, respectively. The levels of Ace-AMP1 in different
transgenic lines correlated with the transcript levels of the transgene. Up to
50% increase in resistance to Blumeria graminis f. sp. tritici was detected in
detached leaf assays. In ears of transgenic wheat inoculated with Neovossia
indica, Ace-AMP1 intensified expression of defense-related genes. Elevated levels
of salicylic acid and of transcripts of phenylalanine ammonia lyase (PAL),
glucanase (PR2) and chitinase (PR3) in the transgenic plants indicated
manifestation of systemic acquired resistance (SAR).

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16906444 [PubMed - indexed for MEDLINE]

1179: Transgenic Res. 2006 Aug;15(4):409-25.

Assessing the potential for unintended effects in genetically modified potatoes
perturbed in metabolic and developmental processes. Targeted analysis of key
nutrients and anti-nutrients.

Shepherd LV, McNicol JW, Razzo R, Taylor MA, Davies HV.

Quality, Health and Nutrition Programme, Scottish Crop Research Institute,
Invergowrie, DD2 5DA, Dundee, Scotland. louise.shepherd@scri.ac.uk

Targeted compositional analysis was carried out on transgenic potato tubers of
either cultivar (cv.) Record or cv. Desirö©e to assess the potential for
unintended effects caused by the genetic modification process. The range of
transgenic lines analysed included those modified in primary carbohydrate
metabolism, polyamine biosynthesis and glycoprotein processing. Controls included
wildtype tubers, tubers produced from plants regenerated through tissue culture
(including a callus phase) and tubers derived from transformation with the 'empty
vector' i.e. no specific target gene included (with the exception of the
kanamycin resistance gene as a selectable marker). Metabolite analysis included
soluble carbohydrates, glycoalkaloids, vitamin C, total nitrogen and fatty acids.
Trypsin inhibitor activity was also assayed. These cover the major compounds
recommended by the OECD in their Consensus Document on Compositional
Considerations for New Varieties of Potatoes: Key Food and Feed Nutrients,
Anti-Nutrients and Toxicants (2002). Data was statistically analysed using
analysis of variance (ANOVA) for individual compounds and, where applicable,
principal component analysis (PCA). In general, targeted compositional analysis
revealed no consistent differences between GM lines and respective controls. No
construct specifically induced unintended effects. Statistically significant
differences between wildtype controls and specific GM lines did occur but
appeared to be random and not associated with any specific construct. Indeed such
significant differences were also found between wildtypes and both tissue culture
derived tubers and tubers derived from transformation with the empty vector. This
raises the possibility that somaclonal variation (known to occur significantly in
potato, depending on genotype) may be responsible for an unknown proportion of
any differences observed between specific GM lines and the wildtype. The most
obvious differences seen in GC-MS profiles were between the two potato varieties
used in the study.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16906442 [PubMed - indexed for MEDLINE]

1180: Transgenic Res. 2006 Aug;15(4):405-7.

Cloned transgenic heart-healthy pork?

Prather RS.

Division of Animal Sciences, University of Missouri-Columbia, 920 East Campus
Drive, E125 Animal Science Research centre, Columbia, MO 65211, USA.
pratherr@missouri.edu

Here I comment on the production and uses of swine that express a humanized fat-1
gene. The gene product is a fatty acid desaturase that converts omega-6 fatty
acids to omega-3 fatty acids. Omega-3 fatty acids have been implicated as being
important for reproductive success, maintaining a healthy cardiovascular system,
sustaining a functional immune system, and even preventing depression and cancer.
The descendants of these hfat-1 transgenic swine will be very useful as models of
the human condition, and if they are permitted to enter the food chain, they may
improve human health.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Review

PMID: 16906441 [PubMed - indexed for MEDLINE]

1181: Transgenic Res. 2006 Aug;15(4):399-404.

Phytohormones and rice crop yield: strategies and opportunities for genetic
improvement.

Sakamoto T.

Field Production Science Center, Graduate School of Agricultural and Life
Sciences, The University of Tokyo, Nishi-Tokyo, Tokyo 188-0002, Japan.
orchardist@fm.a.u-tokyo.ac.jp

To feed an estimated world population of 8.9 billion by 2050, strategies for
increasing grain production must be developed. Several agronomically important
traits for increasing yield, such as plant height, grain number, and leaf
erectness, have recently been characterized in rice (Oryza sativa L.). These
traits are regulated primarily by three phytohormones: gibberellins, cytokinins,
and brassinosteroids. The control of biosynthesis and degradation of these key
phytohormones is discussed in terms of its importance for normal plant growth.
Genes involved in the biosynthesis and regulation of these phytohormones can be
used to develop effective strategies to increase grain yield. Genetic
manipulation of phytohormone-related gene expression is thus a practical strategy
to generate high-yielding transgenic plants through the modification of levels
and profile of endogenous phytohormones.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 16906440 [PubMed - indexed for MEDLINE]

1182: J Plant Physiol. 2007 Jul;164(7):835-41. Epub 2006 Aug 10.

Constitutive expression of an endoplasmic reticulum small heat shock protein
alleviates endoplasmic reticulum stress in transgenic tomato.

Zhao C, Shono M, Sun A, Yi S, Li M, Liu J.

College of Life Science, Shandong Normal University, Jinan, Shandong, PR China.

To explore the function of endoplasmic reticulum-located small heat-shock
proteins (ER-sHSPs) in ER stress, a putative ER-sHSP cDNA (the driven protein was
named as LeHSP21.5 with GenBank accession No. AB026983) was isolated from tomato
(Lycopersicon esculentum). Fractionation of the crude microsomes by isopycnic
sucrose-gradient centrifugation revealed that LeHSP21.5 was distributed on a
density corresponding to the fractions with a higher activity of ER marker
enzyme, suggesting the localization of LeHSP21.5 in the ER. Overexpressing
LeHSP21.5 in transgenic tomato plants (L. esculentum var. Zhongshu 4) greatly
attenuated the lethal effect of tunicamycin on tomato seedlings. Moreover, under
the tunicamycin treatment, transcripts of BiP, PDI and calnexin in transgenic
tomato plants accumulated to a less level than those in non-transgenic tomato
plants. These results suggest that LeHSP21.5 can function to alleviate the
tunicamycin-induced ER stress.

PMID: 16904232 [PubMed - indexed for MEDLINE]

1183: Trends Genet. 2006 Oct;22(10):525-8. Epub 2006 Aug 9.

Shall I compare thee to a GM potato?

Colquhoun IJ, Le Gall G, Elliott KA, Mellon FA, Michael AJ.

Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK.

A fundamental issue in the safety assessment of genetically modified crops is the
question of whether unintentional changes have occurred in the crop plant as a
consequence of the genetic modification. This question was addressed recently by
using a powerful metabolite fingerprinting and metabolite profiling method to
assess whether genetically modified potatoes are substantially similar to their
corresponding conventional cultivars.

Publication Types:
Comparative Study

PMID: 16904227 [PubMed - indexed for MEDLINE]

1184: Biotechnol Lett. 2006 Oct;28(19):1537-44. Epub 2006 Aug 11.

An optimal pooling strategy applied to high-throughput screening for rare
marker-free transformants.

Zhang J, Xiao Q, Li K, Chen M, Chang J, Luo L, Li Y, Liu Y, Shewry PR, He G.

China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory, Huazhong
University of Science and Technology (HUST), Wuhan, Hubei 430074, China.

An optimal pooling system, called Accurate and Fast Target Screening, has been
developed for high-throughput identifying the rare marker-free transformants.
This system can identify targets between 10- and 100-fold more efficiently than
analysis of individual samples. By calculating the efficiency for different
proportions of targets and the optimal group size in a worst case scenario, we
are able to estimate an upper limit for the number of tests that are required.
The application of this system to determine the transgene in an artificially
constructed population of transgenic and non-transgenic wheat lines successfully
identified the 10 positive samples located randomly with 990 negative samples
using only 92 PCR reactions. The same approach was also applied to determine
transgene expression by SDS-PAGE of seed proteins. This system gives unambiguous
positive or negative results and should facilitate marker-free transformation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16902850 [PubMed - indexed for MEDLINE]

1185: Food Addit Contam. 2006 Sep;23(9):876-82.

Development of an innovative immunoassay for CP4EPSPS and Cry1AB genetically
modified protein detection and quantification.

Ermolli M, Prospero A, Balla B, Querci M, Mazzeo A, Van Den Eede G.

Biotechnology & GMOs Unit, Institute for Health and Consumer Protection (IHCP),
European Commission, DG-Joint Research Centre (JRC), Via. E. Fermi 1, 201020
Ispra (VA), Italy. monica.ermolli@jrc.it

An innovative immunoassay, called enzyme-linked immunoabsorbant assay (ELISA)
Reverse, based on a new conformation of the solid phase, was developed. The solid
support was expressly designed to be immersed directly in liquid samples to
detect the presence of protein targets. Its application is proposed in those
cases where a large number of samples have to be screened simultaneously or when
the simultaneous detection of different proteins is required. As a first
application, a quantitative immunoassay for Cry1AB protein in genetically
modified maize was optimized. The method was tested using genetically modified
organism concentrations from 0.1 to 2.0%. The limit of detection and limit of
quantitation of the method were determined as 0.0056 and 0.0168 (expressed as the
percentage of genetically modified organisms content), respectively. A
qualitative multiplex assay to assess the presence of two genetically modified
proteins simultaneously was also established for the case of the Cry1AB and the
CP4EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) present in genetically
modified maize and soy, respectively.

PMID: 16901856 [PubMed - indexed for MEDLINE]

1186: Plant Foods Hum Nutr. 2006 Sep;61(3):145-50.

Effect of cultivar, year grown, and cropping system on the content of tocopherols
and tocotrienols in grains of hulled and hulless barley.

Ehrenbergerovö¡ J, Belcrediovö¡ N, Prö½ma J, Vaculovö¡ K, Newman CW.

Mendel University of Agriculture and Forestry, Brno, Czech Republic.
ehren@mendelu.cz

In a three-year period (2000-2002) total tocols (tocopherols and tocotrienols),
content of vitamin E and its isomers (alpha-, beta+gamma-, delta-tocopherols and
tocotrienols) were assessed in grain of 13 barley genotypes. The highest content
of tocols (60.3-67.6 mg kg(-1)) and content of vitamin E (Vitamin E
equivalent-18.0-20.1 mg kg(-1)) were determined in the waxy varieties Wanubet,
Wabet, and Washonubet. Standard varieties, i.e. of a malting type (Krona and
Kompakt), had statistically significantly lower content of tocols (49.9 and 53.6
mg kg(-1)) and vitamin E (15.7-16.1 mg kg(-1)) compared to the waxy varieties.
The hulless waxy variety Washonubet had statistically significantly higher total
content of tocols (67.6 mg kg(-1)) and alpha- tocotrienols isomer (42.1 mg
kg(-1)) versus all other genotypes in the set. Chemical treatment and
fertilization statistically significantly increased the content of tocols (by 4.7
mg kg(-1)), vitamin E (by 1.9 mg kg(-1)), isomer alpha-tocopherol (by 0.9 mg
kg(-1)) and isomer alpha- tocotrienols (by 3.3 mg kg(-1)). The average values of
alpha-tocopherols and alpha-tocotrienols in the set were 6.7 mg kg(-1) and 29.7
mg kg(-1), respectively. Some of the reciprocal lines created by us from the
malting and waxy varieties are suitable for food use for high contents of all
tocopherols and alpha-tocotrienols.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16900405 [PubMed - indexed for MEDLINE]

1187: Plant Mol Biol. 2006 Sep;62(1-2):15-28. Epub 2006 Aug 10.

Nature of stress and transgene locus influences transgene expression stability in
barley.

Meng L, Ziv M, Lemaux PG.

Department of Plant and Microbial Biology, University of California, 111 Koshland
Hall, Berkeley , CA 94720, USA.

Stress and the nature of the transgene locus can affect transgene expression
stability. These effects were studied in two, stably expressing, T6 populations
of barley (Hordeum vulgare): bombardment-mediated, multi-copy lines with
ubiquitin-driven bar and uidA or single-copy lines from Ds-mediated gene delivery
with ubiquitin-driven bar alone. Imposing the environmental stresses, water and
nutrient deprivation and heat shock, did not reproducibly affect transgene
expression stability; however, high frequencies of heritable transcriptional gene
silencing (TGS) occurred following in vitro culture after six generations of
stable expression in the multi-copy subline, T3#30, but not in the other lines
studied. T3#30 plants with complete TGS had epigenetic modification patterns
exactly like those in an identical sibling subline, T3#31, which had significant
reduction in transgene expression in the T3 generation and was completely
transcriptionally silenced in the absence of imposed stresses in the T6
generation. Complete TGS in T3#30 plants correlated with methylation in the 5'UTR
and intron of the ubi1 promoter complex and condensation of chromatin around the
transgenes; DNA methylation likely occurred prior to chromatin condensation.
Partial TGS in T3#30 also correlated with methylation of the ubi1 promoter
complex, as occurred with complete TGS. T3#30 has a complex transgene structure
with inverted repeat transgene fragments and a 3'-LTR from a barley
retrotransposon, and therefore the transgene locus itself may affect its tendency
to silence after in vitro culture and transgene silencing might result from host
defense mechanisms activated by changes in plant developmental programming and/or
stresses imposed during in vitro growth.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16900326 [PubMed - indexed for MEDLINE]

1188: Genetica. 2007 May;130(1):61-72. Epub 2006 Aug 10.

Phenotypic effect of substitution of allelic variants for a histone H1 subtype
specific for growing tissues in the garden pea (Pisum sativum L.).

Bogdanova VS, Kosterin OE, Berdnikov VA.

Institute of Cytology & Genetics, Acad Lavrentiev Ave 10, Novosibirsk, Russia.
vera@bionet.nsc.ru

In pea, subtype H1-7 of histone H1 is specific for young actively growing tissues
and disappears from chromatin of mature tissues. We sequenced the alleles coding
for three main variants, numbered according to the increase of the
electrophoretic mobility. Allele 1 differs from the most common allele 2 by eight
nucleotide substitutions, two of them associated with amino acid replacements,
His->Tyr in the globular domain and Ala->Val in the C-terminal domain. Allele 3
differs from alleles 1 and 2 by a 24-bp deletion in the part coding for the
C-terminal domain. In three greenhouse experiments, we compared quantitative
traits in nearly isogenic lines differing by these H1-7 variants. In experiment
1, three lines bearing either of the three allelic variants were compared, the
other experiments involved pairs of lines bearing variants 1 and 3. In all
experiments, statistically significant differences between the lines were
registered, mostly related to the plant size. The most prominent effect was
associated with plant growth dynamics. Plants of line 3, carrying the 8-amino
acid deletion in histone H1-7, on average grew slower. In two experiments, the
differences of the mean stem length persisted throughout plant growth while in
experiment 2 differences disappeared upon maturity. The H1-7 subtype is supposed
to be related to maintenance of chromatin state characteristic for cell growth
and division.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16900316 [PubMed - indexed for MEDLINE]

1189: Nature. 2006 Aug 10;442(7103):705-8.

Comment in:
Nature. 2006 Aug 10;442(7103):635-6.

Sub1A is an ethylene-response-factor-like gene that confers submergence tolerance
to rice.

Xu K, Xu X, Fukao T, Canlas P, Maghirang-Rodriguez R, Heuer S, Ismail AM,
Bailey-Serres J, Ronald PC, Mackill DJ.

Department of Plant Pathology, University of California, Davis, California 95616,
USA.

Most Oryza sativa cultivars die within a week of complete submergence--a major
constraint to rice production in south and southeast Asia that causes annual
losses of over US 1 billion dollars and affects disproportionately the poorest
farmers in the world. A few cultivars, such as the O. sativa ssp. indica cultivar
FR13A, are highly tolerant and survive up to two weeks of complete submergence
owing to a major quantitative trait locus designated Submergence 1 (Sub1) near
the centromere of chromosome 9 (refs 3, 4, 5-6). Here we describe the
identification of a cluster of three genes at the Sub1 locus, encoding putative
ethylene response factors. Two of these genes, Sub1B and Sub1C, are invariably
present in the Sub1 region of all rice accessions analysed. In contrast, the
presence of Sub1A is variable. A survey identified two alleles within those
indica varieties that possess this gene: a tolerance-specific allele named
Sub1A-1 and an intolerance-specific allele named Sub1A-2. Overexpression of
Sub1A-1 in a submergence-intolerant O. sativa ssp. japonica conferred enhanced
tolerance to the plants, downregulation of Sub1C and upregulation of Alcohol
dehydrogenase 1 (Adh1), indicating that Sub1A-1 is a primary determinant of
submergence tolerance. The FR13A Sub1 locus was introgressed into a widely grown
Asian rice cultivar using marker-assisted selection. The new variety maintains
the high yield and other agronomic properties of the recurrent parent and is
tolerant to submergence. Cultivation of this variety is expected to provide
protection against damaging floods and increase crop security for farmers.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16900200 [PubMed - indexed for MEDLINE]

1190: J Environ Qual. 2006 Aug 9;35(5):1633-58. Print 2006 Sep-Oct.

The current status and environmental impacts of glyphosate-resistant crops: a
review.

Cerdeira AL, Duke SO.

Brazilian Department of Agriculture, Agricultural Research Service,
Embrapa/Environment, C.P. 69, Jaguariuna-SP-13820-000, Brazil.

Glyphosate [N-(phosphonomethyl) glycine]-resistant crops (GRCs), canola (Brassica
napus L.), cotton (Gossypium hirsutum L.), maize (Zea mays L.), and soybean
[Glycine max (L.) Merr.] have been commercialized and grown extensively in the
Western Hemisphere and, to a lesser extent, elsewhere. Glyphosate-resistant
cotton and soybean have become dominant in those countries where their planting
is permitted. Effects of glyphosate on contamination of soil, water, and air are
minimal, compared to some of the herbicides that they replace. No risks have been
found with food or feed safety or nutritional value in products from currently
available GRCs. Glyphosate-resistant crops have promoted the adoption of reduced-
or no-tillage agriculture in the USA and Argentina, providing a substantial
environmental benefit. Weed species in GRC fields have shifted to those that can
more successfully withstand glyphosate and to those that avoid the time of its
application. Three weed species have evolved resistance to glyphosate in GRCs.
Glyphosate-resistant crops have greater potential to become problems as volunteer
crops than do conventional crops. Glyphosate resistance transgenes have been
found in fields of canola that are supposed to be non-transgenic. Under some
circumstances, the largest risk of GRCs may be transgene flow (introgression)
from GRCs to related species that might become problems in natural ecosystems.
Glyphosate resistance transgenes themselves are highly unlikely to be a risk in
wild plant populations, but when linked to transgenes that may impart fitness
benefits outside of agriculture (e.g., insect resistance), natural ecosystems
could be affected. The development and use of failsafe introgression barriers in
crops with such linked genes is needed.

Publication Types:
Review

PMID: 16899736 [PubMed - indexed for MEDLINE]

1191: Plant Mol Biol. 2006 Nov;62(4-5):547-59. Epub 2006 Aug 1.

DEA1, a circadian- and cold-regulated tomato gene, protects yeast cells from
freezing death.

Weyman PD, Pan Z, Feng Q, Gilchrist DG, Bostock RM.

Department of Plant Pathology, University of California, One Shields Ave, Davis,
CA 95616, USA.

Cold and freezing damage to plants can be mitigated by inducible factors during
an acclimation period. DEA1 is a circadian-regulated tomato (Solanum
lycopersicum) gene with sequence similarity to EARLI1, an Arabidopsis thaliana
gene that confers cold protection. To investigate whether DEA1 was responsive to
environmental variables such as cold, cold-treated tomatoes were analyzed for
DEA1 expression. DEA1 transcript accumulated in response to cold, and the
rapidity of the cold-induced transcript accumulation was regulated by the
circadian rhythm. To test whether DEA1 could protect cells from freezing damage,
we transformed the yeast, Pichia pastoris, with an inducible DEA1 construct.
Yeast cells transformed with the gene survived freezing at a significantly higher
rate than control strains and a strain expressing the LacZ gene. Transgenic
tomato plants over-expressing or knocking down DEA1 transcript levels did not
have an altered phenotype with respect to cold- or pathogen-susceptibility
relative to control plants.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16897467 [PubMed - indexed for MEDLINE]

1192: Planta. 2006 Dec;225(1):153-64. Epub 2006 Jul 29.

Genetic and transgenic perturbations of carbon reserve production in Arabidopsis
seeds reveal metabolic interactions of biochemical pathways.

Lin Y, Ulanov AV, Lozovaya V, Widholm J, Zhang G, Guo J, Goodman HM.

Department of Crop Sciences, University of Illinois, Urbana, IL 61801, USA.
yunlin@uiuc.edu

The biosynthesis of seed oil and starch both depend on the supply of carbon from
the maternal plant. The biochemical interactions between these two pathways are
not fully understood. In the Arabidopsis mutant shrunken seed 1 (sse1)/pex16, a
reduced rate of fatty acid synthesis leads to starch accumulation. To further
understand the metabolic impact of the decrease in oil synthesis, we compared
soluble metabolites in sse1 and wild type (WT) seeds. Sugars, sugar phosphates,
alcohols, pyruvate, and many other organic acids accumulated in sse1 seeds as a
likely consequence of the reduced carbon demand for lipid synthesis. The enlarged
pool size of hexose-P, the metabolites at the crossroad of sugar metabolism,
glycolysis, and starch synthesis, was likely a direct cause of the increased flow
into starch. Downstream of glycolysis, more carbon entered the TCA cycle as an
alternative to the fatty acid pathway, causing the total amount of TCA cycle
intermediates to rise while moving the steady state of the cycle away from
fumarate. To convert the excess carbon metabolites into starch, we introduced the
Escherichia coli starch synthetic enzyme ADP-glucose pyrophosphorylase (AGPase)
into sse1 seeds. Expression of AGPase enhanced net starch biosynthesis in the
mutant, resulting in starch levels that reached 37% of seed weight. However,
further increases above this level were not achieved and most of the carbon
intermediates remained high in comparison with the WT, indicating that additional
mechanisms limit starch deposition in Arabidopsis seeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16896794 [PubMed - indexed for MEDLINE]

1193: Proc Natl Acad Sci U S A. 2006 Aug 15;103(33):12215-6. Epub 2006 Aug 7.

Comment on:
Proc Natl Acad Sci U S A. 2006 Aug 15;103(33):12329-34.

Agriculture: the selector of improbable mutations.

Gressel J, Levy AA.

Department of Plant Sciences, Weizmann Institute of Science, Rehovot 76100,
Israel. jonathan.gressel@weizmann.ac.il

Publication Types:
Comment

PMID: 16894157 [PubMed - indexed for MEDLINE]

1194: Plant Physiol Biochem. 2006 May-Jun;44(5-6):380-6. Epub 2006 Jun 22.

rHsp90 gene expression in response to several environmental stresses in rice
(Oryza sativa L.).

Liu D, Zhang X, Cheng Y, Takano T, Liu S.

Alkali Soil Natural Environmental Science Center (ASNESC), Stress Molecular
Biology Laboratory, Northeast Forestry University, Harbin 150040, PR China.

In this study, the gene for a rice (Oryza sativa L.) 90 kDa heat shock protein
(rHsp90, GenBank accession no. AB037681) was identified by screening rice root
cDNAs that were up-regulated under carbonate (NaHCO(3)) stress using the method
of differential display, and cloned. The open-reading-frame of rHsp90-cDNA was
predicted to encode a protein containing 810 amino acids, which showed high
similarity to proteins in Hordeum vulgare (accession no. X67960) and Catharathus
roseus (accession no. L14594). Further studies showed that rHsp90 mRNA
accumulated following exposure to several abiotic stresses, including salts
(NaCl, NaHCO(3) and Na(2)CO(3)), desiccation (using polyethylene glycol), high pH
(8.0 and 11.0) and high temperature (42 and 50 degrees C). Yeast (Saccharomyces
cerevisiae) over-expressing rHsp90 exhibited greater tolerance to NaCl,
Na(2)CO(3) and NaHCO(3) and tobacco seedlings over-expressing rHsp90 could
tolerate salt concentrations as high as 200 mM NaCl, whereas untransformed
control seedlings couldn't. These results suggest that rHsp90 plays an important
role in multiple environmental stresses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16889974 [PubMed - indexed for MEDLINE]

1195: Plant Biol (Stuttg). 2006 Sep;8(5):579-86. Epub 2006 Aug 1.

Effects of cytokinin production under two SAG promoters on senescence and
development of tomato plants.

Swartzberg D, Dai N, Gan S, Amasino R, Granot D.

Institute of Plant Sciences, Agricultural Research Organization, The Volcani
Center, P.O. Box 6, Bet Dagan, 50250, Israel.

Two promoters of senescence-associated ARABIDOPSIS genes, SAG12 and SAG13, were
used in tomato plants to express IPT that catalyzes the rate-limiting step in
cytokinin biosynthesis. Expression of these heterologous promoters in tomato
plants was analyzed using the reporter gene beta-glucuronidase. Both promoters
are expressed in tomato leaves in a manner similar to their expression in
ARABIDOPSIS plants. The SAG12 promoter is very specific to senescing leaves,
whereas the SAG13 promoter is expressed in mature leaves prior to the onset of
visible senescence and its expression increases in senescing leaves. Expression
of both promoters in tomato tissues other than leaves was very low . IPT
expressed under the control of SAG12 and SAG13 promoters ( PSAG12::IPT and
PSAG13::IPT, respectively) resulted in suppression of leaf senescence and
advanced flowering, as well as in a slight increase in fruit weight and fruit
total soluble solids (TSS). However, expression of PSAG13::IPT also led to stem
thickening, short internodal distances and loss of apical dominance. In contrast
to the autoregulation of PSAG12::IPT, PSAG13::IPT is expressed at higher levels
in mature leaves. This difference is likely due to PSAG13::IPT exhibiting two
phases of expression - a senescence-independent expression prior to the onset of
senescence that is not subjected to autoregulation by cytokinin, and enhanced
expression throughout senescence which is autoregualted by cytokinin. This
moderate different autoregulated behavior of PSAG12::IPT and PSAG13::IPT markedly
influenced plant development, emphasizing the biological effects of cytokinin in
addition to senescence inhibition.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16883480 [PubMed - indexed for MEDLINE]

1196: Plant Biol (Stuttg). 2006 Sep;8(5):723-30. Epub 2006 Aug 1.

Erratum in:
Plant Biol (Stuttg). 2006 Nov;8(6):861-3.

Correlated expression of gfp and Bt cry1Ac gene facilitates quantification of
transgenic hybridization between Brassicas.

Shen BC, Stewart CN Jr, Zhang MQ, Le YT, Tang ZX, Mi XC, Wei W, Ma KP.

Laboratory of Quantitative Vegetation Ecology, Institute of Botany, Chinese
Academy of Science, Beijing 100093, China.

Gene flow from transgenic oilseed rape (BRASSICA NAPUS) might not be avoidable,
thus, it is important to detect and quantify hybridization events with its
relatives in real time. Data are presented showing the correlation between
genetically linked green fluorescent protein (GFP) with BACILLUS THURINGIENSIS
(Bt) CRY1AC gene expression in hybrids formed between transgenic B. NAPUS
"Westar" and a wild Chinese accession of wild mustard (B. JUNCEA) and
hybridization between transgenic B. NAPUS and a conspecific Chinese landrace
oilseed rape. Hybrids were obtained either by spontaneous hybridization in the
field or by hand-crossing in a greenhouse. In all cases, transgenic hybrids were
selected by GFP fluorescence among seedlings originating from seeds harvested
from B. JUNCEA and the Chinese oilseed rape plants. Transgenicity was confirmed
by PCR detection of transgenes. GFP fluorescence was easily and rapidly detected
in the hybrids under greenhouse and field conditions. Results showed that both
GFP fluorescence and Bt protein synthesis decreased as either plant or leaf aged,
and GFP fluorescence intensity was closely correlated with Bt protein
concentration during the entire vegetative lifetime in hybrids. These findings
allow the use of GFP fluorescence as an accurate tool to detect gene-flow in time
in the field and to conveniently estimate BT CRY1AC expression in hybrids
on-the-plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16883477 [PubMed - indexed for MEDLINE]

1197: Protein Sci. 2006 Sep;15(9):2040-50. Epub 2006 Aug 1.

The refolding of different alpha-fetoprotein variants.

Leong SS, Middelberg AP.

Centre for Biomolecular Engineering, The University of Queensland, St. Lucia,
Australia. ssjl2@cam.ac.uk

The effect of glycosylation on AFP foldability was investigated by parallel
quantitative and qualitative analyses of the refolding of glycosylated and
nonglycosylated AFP variants. Both variants were successfully refolded by
dialysis from the denatured-reduced state, attaining comparable "refolded peak"
profiles and refolding yields as determined by reversed-phase HPLC analysis. Both
refolded variants also showed comparable spectroscopic fingerprints to each other
and to their native counterparts, as determined by circular dichroism
spectroscopy. Inclusion body-derived AFP was also readily refolded via dilution
under the same redox conditions as dialysis refolding, showing comparable
circular dichroism fingerprints as native nonglycosylated AFP. Quantitative
analyses of inclusion body-derived AFP showed sensitivity of AFP aggregation to
proteinaceous and nonproteinaceous inclusion body contaminants, where refolding
yields increased with increasing AFP purity. All of the refolded AFP variants
showed positive responses in ELISA that corresponded with the attainment of a
bioactive conformation. Contrary to previous reports that the denaturation of
cord serum AFP is an irreversible process, these results clearly show the
reversibility of AFP denaturation when refolded under a redox-controlled
environment, which promotes correct oxidative disulfide shuffling. The successful
refolding of inclusion body-derived AFP suggests that fatty acid binding may not
be required for the attainment of a rigid AFP tertiary structure, contrary to
earlier studies. The overall results from this work demonstrate that foldability
of the AFP molecule from its denatured-reduced state is independent of its
starting source, the presence or absence of glycosylation and fatty acids, and
the refolding method used (dialysis or dilution).

PMID: 16882993 [PubMed - indexed for MEDLINE]

1198: Environ Health Perspect. 2006 Aug;114(8):1154-7.

Digestion assays in allergenicity assessment of transgenic proteins.

Herman RA, Storer NP, Gao Y.

Dow AgroSciences LLC, Indianapolis, Indiana 46268, USA. raherman@dow.com

The food-allergy risk assessment for transgenic proteins expressed in crops is
currently based on a weight-of-evidence approach that holistically considers
multiple lines of evidence. This approach recognizes that no single test or
property is known to distinguish allergens from nonallergens. The stability of a
protein to digestion, as predicted by an in vitro simulated gastric fluid assay,
currently is used as one element in the risk assessment process. A review of the
literature on the use of the simulated gastric fluid assay to predict the
allergenic status of proteins suggests that more extensive kinetic studies with
well-characterized reference proteins are required before the predictive value of
this assay can be adequately judged.

Publication Types:
Review

PMID: 16882518 [PubMed - indexed for MEDLINE]

1199: EMBO Rep. 2006 Aug;7(8):750-3.

Cisgenic plants are similar to traditionally bred plants: international
regulations for genetically modified organisms should be altered to exempt
cisgenesis.

Schouten HJ, Krens FA, Jacobsen E.

Plant Research International, Wageningen University and Research Centre in the
Netherlands. henk.schouten@wur.nl

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16880817 [PubMed - indexed for MEDLINE]

1200: Proc Natl Acad Sci U S A. 2006 Aug 8;103(32):12197-202. Epub 2006 Jul 31.

Manipulation of Arabidopsis fatty acid amide hydrolase expression modifies plant
growth and sensitivity to N-acylethanolamines.

Wang YS, Shrestha R, Kilaru A, Wiant W, Venables BJ, Chapman KD, Blancaflor EB.

Plant Biology Division, Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway,
Ardmore, OK 73401, USA.

In vertebrates, the endocannabinoid signaling pathway is an important lipid
regulatory pathway that modulates a variety of physiological and behavioral
processes. N-Acylethanolamines (NAEs) comprise a group of fatty acid derivatives
that function within this pathway, and their signaling activity is terminated by
an enzyme called fatty acid amide hydrolase (FAAH), which hydrolyzes NAEs to
ethanolamine and their corresponding free fatty acids. Bioinformatic approaches
led to the identification of plant homologues of FAAH that are capable of
hydrolyzing NAEs in vitro. To better understand the role of NAEs in plants, we
identified T-DNA knockouts to Arabidopsis FAAH (AtFAAH; At5g64440) and generated
plants overexpressing AtFAAH. Here we show that seeds of AtFAAH knockouts had
elevated levels of endogenous NAEs, and seedling growth was hypersensitive to
exogenously applied NAE. On the other hand, seeds and seedlings of AtFAAH
overexpressors had lower endogenous NAE content, and seedlings were less
sensitive to exogenous NAE. Moreover, AtFAAH overexpressors displayed enhanced
seedling growth and increased cell size. AtFAAH expression and FAAH catalytic
activity increased during seed germination and seedling growth, consistent with
the timing of NAE depletion during seedling establishment. Collectively, our
results show that AtFAAH is one, but not the only, modulator of endogenous NAE
levels in plants, and that NAE depletion likely participates in the regulation of
plant growth.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16880402 [PubMed - indexed for MEDLINE]

1201: Proc Natl Acad Sci U S A. 2006 Aug 8;103(32):11856-61. Epub 2006 Jul 31.

A viral resistance gene from common bean functions across plant families and is
up-regulated in a non-virus-specific manner.

Seo YS, Rojas MR, Lee JY, Lee SW, Jeon JS, Ronald P, Lucas WJ, Gilbertson RL.

Department of Plant Pathology and Section of Plant Biology, University of
California, Davis, CA 95616, USA.

Genes involved in a viral resistance response in common bean (Phaseolus vulgaris
cv. Othello) were identified by inoculating a geminivirus reporter (Bean dwarf
mosaic virus expressing the green fluorescent protein), extracting RNA from
tissue undergoing the defense response, and amplifying sequences with degenerate
R gene primers. One such gene (a TIR-NBS-LRR gene, RT4-4) was selected for
functional analysis in which transgenic Nicotiana benthamiana were generated and
screened for resistance to a range of viruses. This analysis revealed that RT4-4
did not confer resistance to the reporter geminivirus; however, it did activate a
resistance-related response (systemic necrosis) to seven strains of Cucumber
mosaic virus (CMV) from pepper or tomato, but not to a CMV strain from common
bean. Of these eight CMV strains, only the strain from common bean systemically
infected common bean cv. Othello. Additional evidence that RT4-4 is a CMV R gene
came from the detection of resistance response markers in CMV-challenged leaves
of RT4-4 transgenic plants, and the identification of the CMV 2a gene product as
the elicitor of the necrosis response. These findings indicate that RT4-4
functions across two plant families and is up-regulated in a non-virus-specific
manner. This experimental approach holds promise for providing insights into the
mechanisms by which plants activate resistance responses against pathogens.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16880399 [PubMed - indexed for MEDLINE]

1202: Cell Microbiol. 2007 Jan;9(1):120-30. Epub 2006 Jul 31.

Activity of HIV entry and fusion inhibitors expressed by the human vaginal
colonizing probiotic Lactobacillus reuteri RC-14.

Liu JJ, Reid G, Jiang Y, Turner MS, Tsai CC.

Washington National Primate Research Center, University of Washington, Seattle,
WA 98195, USA. liuj@u.washington.edu

Novel therapeutic approaches are needed to combat the rapid increase in HIV
sexual transmission in women. The probiotic organism Lactobacillus reuteri RC-14
which safely colonizes the human vagina and prevents microbial infections, has
been genetically modified to produce anti-HIV proteins which were capable of
blocking the three main steps of HIV entry into human peripheral blood
mononuclear cells. The HIV entry or fusion inhibitors were fused to the native
expression and secretion signals of BspA, Mlp or Sep in L. reuteri RC-14 and the
expression cassettes were stably inserted into the chromosome. L. reuteri RC-14
expressed the HIV inhibitors in cell wall-associated and secreted forms. L.
reuteri RC-14 expressing CD4D1D2-antibody-like fusion proteins were able to bind
single or dual tropic coreceptor-using HIV-1 primary isolates. This is the first
study to show that a well-documented and proven human vaginal probiotic strain
can express potent functional viral inhibitors, which may potentially lower the
sexual transmission of HIV.

PMID: 16879452 [PubMed - indexed for MEDLINE]

1203: J Exp Bot. 2006;57(12):3007-18. Epub 2006 Jul 26.

Overexpression of a bacterial 1-deoxy-D-xylulose 5-phosphate synthase gene in
potato tubers perturbs the isoprenoid metabolic network: implications for the
control of the tuber life cycle.

Morris WL, Ducreux LJ, Hedden P, Millam S, Taylor MA.

Quality, Health and Nutrition, Scottish Crop Research Institute, Invergowrie,
Dundee DD2 5DA, UK.

Potato tubers were engineered to express a bacterial gene encoding
1-deoxy-D-xylulose 5-phosphate synthase (DXS) in order to investigate the effects
of perturbation of isoprenoid biosynthesis. Twenty-four independent transgenic
lines out of 38 generated produced tubers with significantly elongated shape that
also exhibited an early tuber sprouting phenotype. Expression analysis of nine
transgenic lines (four exhibiting the phenotype and five showing a wild-type
phenotype) demonstrated that the phenotype was strongly associated with dxs
expression. At harvest, apical bud growth had already commenced in dxs-expressing
tubers whereas in control lines no bud growth was evident until dormancy was
released after 56-70 d of storage. The initial phase of bud growth in dxs tubers
was followed by a lag period of approximately 56 d, before further elongation of
the developing sprouts could be detected. Thus dxs expression results in the
separation of distinct phases in the dormancy and sprouting processes. In order
to account for the sprouting phenotype, the levels of plastid-derived isoprenoid
growth regulators were measured in transgenic and control tubers. The major
difference measured was an increase in the level of trans-zeatin riboside in
tubers at harvest expressing dxs. Additionally, compared with controls, in some
dxs-expressing lines, tuber carotenoid content increased approximately 2-fold,
with most of the increase accounted for by a 6-7-fold increase in phytoene.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16873449 [PubMed - indexed for MEDLINE]

1204: Phytochemistry. 2006 Sep;67(17):1895-906. Epub 2006 Jul 25.

Tomato cytochrome P450 CYP734A7 functions in brassinosteroid catabolism.

Ohnishi T, Nomura T, Watanabe B, Ohta D, Yokota T, Miyagawa H, Sakata K, Mizutani
M.

Institute for Chemical Research, Kyoto University, Gokasyo, Uji, Kyoto 611-0011,
Japan.

Several cytochrome P450 monooxygenases (P450s) catalyze essential oxidative
reactions in brassinosteroid (BR) biosynthesis as well as in BR catabolism;
however, only limited information exists on the P450s involved in the BR
catabolic pathway. Here, we report the characterization of two P450 mRNAs,
CYP734A7 and CYP734A8, from Lycopersicon esculentum. These P450s show high
homology with Arabidopsis CYP734A1/BAS1 (formerly CYP72B1), which inactivates BRs
via C-26 hydroxylation. Transgenic tobacco plants that constitutively
overexpressed CYP734A7 showed an extreme dwarf phenotype similar to BR
deficiency. Quantitative gas chromatography-mass spectrometry analysis of
endogenous BRs in the transgenic plants showed that the levels of castasterone
and 6-deoxocastasterone significantly decreased in comparison with those in
wild-type plants. By measuring the Type I substrate-binding spectra using
recombinant CYP734A7, the dissociation constants for castasterone, brassinolide,
and 6-deoxocastasterone were determined to be 6.7, 12, and 12 microM,
respectively. In an in vitro assay, CYP734A7 was confirmed to metabolize
castasterone to 26-hydroxycastasterone. In addition, 28-norcastasterone and
brassinolide were converted to the hydroxylated products. The expression of
CYP734A7 and CYP734A8 genes in tomato seedlings was upregulated by exogenous
application of bioactive BRs. These results indicated that CYP734A7 is a C-26
hydroxylase of BRs and is likely involved in BR catabolism in tomato. The
presence of the CYP734A subfamily in various plant species suggests that
oxidative inactivation of BRs by these proteins is a widespread phenomenon in
plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16872648 [PubMed - indexed for MEDLINE]

1205: Biochem Biophys Res Commun. 2006 Sep 8;347(4):1030-8. Epub 2006 Jul 14.

Regulation of ABA level and water-stress tolerance of Arabidopsis by ectopic
expression of a peanut 9-cis-epoxycarotenoid dioxygenase gene.

Wan XR, Li L.

Guangdong Key Lab of Biotechnology for Plant Development, College of Life
Science, South China Normal University, Guangzhou 510631, China.

The oxidative cleavage of cis-epoxycarotenoids catalyzed by 9-cis-epoxycarotenoid
dioxygenase (NCED) is considered to be the rate-limiting step in abscisic acid
(ABA) biosynthesis. Here we demonstrate that the expression of AhNCED1 gene in
peanut plants is significantly up-regulated by dehydration and high salinity. The
AhNCED1 transcript and endogenous ABA both accumulate predominantly in leaves and
stems of peanut in response to dehydration. The considerable up-regulation of
AhNCED1 expression by exogenous application of ABA suggests a positive feedback
control of ABA biosynthesis in peanut. NAA at high concentration increases ABA
biosynthesis in peanut plants through up-regulation of the AhNCED1 gene
expression. The constitutive expression of the AhNCED1 gene in wild-type
Arabidopsis results in an increase of ABA accumulation in transgenic plants in
response to drought stress. Ectopic expression of AhNCED1 gene in 129B08/nced3
mutant Arabidopsis (with impaired AtNCED3 gene involved in ABA biosynthesis under
water stress) driven by the AtNCED3 promoter restores its ability to accumulate
ABA during drought stress, and reverts its hypersensitivity to nonionic osmotic
stress and soil drought. These results indicate that the expression of AhNCED1
gene plays an important role in the regulation of ABA level during water stress,
and that water-stress tolerance of Arabidopsis plants can be improved by ectopic
expression of the AhNCED1 gene causing accumulation of endogenous ABA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16870153 [PubMed - indexed for MEDLINE]

1206: Lett Appl Microbiol. 2006 Aug;43(2):215-21.

Transformation of Acinetobacter sp. BD413 with DNA from commercially available
genetically modified potato and papaya.

Iwaki M, Arakawa Y.

Department of Bacterial Pathogenesis and Infection Control, National Institute of
Infectious Diseases, Tokyo, Japan. miwaki@nih.go.jp

AIM: To estimate the likelihood of transfer of kanamycin-resistance gene (nptII)
from commercially available genetically modified (GM) plants. METHODS AND
RESULTS: Acinetobacter sp. BD413 carrying a plasmid containing an inactivated
nptII gene was treated with DNA derived from GM potato and GM papaya.
Kanamycin-resistant transformants were obtained at a frequency of 10-30
microg(-1) DNA. Calculation of the results suggested that 6-9 x 10(4) molecules
of genomic DNA from GM plants were needed to obtain one transformant. However,
such transformation events were not detectable in the absence of the plasmid in
the host strain. CONCLUSIONS: Acinetobacter sp. BD413 was transformed with DNA
derived from GM potato and GM papaya, in the presence of an inactivated nptII
gene on a plasmid. However, the frequency of such events in the natural
environment on wild-type strains, while evidently low, remains unknown.
SIGNIFICANCE AND IMPACT OF THE STUDY: Our results may help to evaluate potential
risks associated with the use of antibiotic-resistance determinants as genetic
markers in GM plants. Complete risk assessment must consider factors other than
transformation frequency alone, including the natural background of antibiotic
resistance present in bacterial populations, and the spectrum and clinical use of
the antimicrobial agents in question.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16869908 [PubMed - indexed for MEDLINE]

1207: Reprod Domest Anim. 2006 Aug;41(4):260-7.

Erratum in:
Reprod Domest Anim. 2006 Oct;41(5):477.

Biosecurity and the various types of embryos transferred.

Thibier M.

Ministö¨re de l'Agriculture et de la Pöªche, Paris, France.
michel.thibier@agriculture.gouv.fr

The aim of the present paper was to review some features related to the risk
analysis of three types of embryos to be transferred, namely the in vivo derived,
the in vitro produced and the cloned ones. For in vivo-collected embryos, a
considerable number of experiments and scientific investigations have been
performed and hundreds of thousands of embryos are transferred annually with no
contamination of associated diseases. Provided that the code of practice such as
that published by the International Embryo Transfer Society is strictly followed
by the embryo transfer practitioners, the statement made some 17 years ago saying
that the in vivo-derived embryo transfer was the safest way of exchanging genes
remains entirely true, thanks to the professionalism of the embryo transfer
industry. For the in vitro-produced embryos, some particular rules have to be
followed because of specific risks for some pathogens to strongly adhere to the
zona pellucida of such embryos. There are some means to monitor and control those
effects, and the transfer of in vitro-produced embryos can also be a very safe
way to exchange genes around the world. The third type of embryos, the cloned
ones, is a quite different category and the risk analysis to be soundly made
still needs a lot of investigations so as to characterize the potential risks if
there are, in terms not only of disease transmission but also in terms of public
health, zoonotic risks as well as those related to quality and safety of food.
The problem in this regard, is more directly addressed for offspring of clones
than to the cloned embryos themselves. Published data on this issue are
increasing in numbers so that progress in that area is expected in the few years
to come.

Publication Types:
Review

PMID: 16869879 [PubMed - indexed for MEDLINE]

1208: Shokuhin Eiseigaku Zasshi. 2006 Jun;47(3):111-4.

A detection method of CryIAc protein for identifying genetically modified rice
using the lateral flow strip assay.

Akiyama H, Watanabe T, Kikuchi H, Sakata K, Tokishita S, Hayashi Y, Hino A,
Teshima R, Sawada J, Maitani T.

National Institute of Health Sciences: 1-18-1, Kamiyoga, Setagaya-ku, Tokyo
158-8501, Japan.

We examined the lateral flow strip assay for identifying unauthorized genetically
modified (GM) rice. The GM rice expresses the Bacillus thuringiensis (Bt) toxin,
CryIAc protein, which confers tolerance to insects. The recombinant CryIAc
protein was prepared from the inclusion bodies of an E. coli. strain into which
the CryIAc gene had been inserted, using gel filtration chromatography. The
lateral flow strip assay for the identification of GM cotton which also expresses
CryIAc protein, was applied to unpolished rice and polished rice spiked with
recombinant CryIAc protein. The spiked recombinant CryIAc protein was clearly
detected at the level of 0.012 microg/g in both the unpolished and polished rice.
After loading of the extract on the strip, a 60 -minute stand time is necessary
to clearly detect CryIAc protein. The detection limit was approximately 12 ng
CryIAc protein per gram of rice. These results suggest that the lateral flow
strip assay for GM cotton can be used to detect CryIAc protein expressed in GM
rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16862988 [PubMed - indexed for MEDLINE]

1209: Mol Genet Genomics. 2006 Oct;276(4):351-68. Epub 2006 Jul 22.

Seed-specific expression of sesame microsomal oleic acid desaturase is controlled
by combinatorial properties between negative cis-regulatory elements in the
SeFAD2 promoter and enhancers in the 5'-UTR intron.

Kim MJ, Kim H, Shin JS, Chung CH, Ohlrogge JB, Suh MC.

School of Life Sciences and Biotechnology, Korea University, Seoul, 136-701,
South Korea.

The regulation of genes involved in primary lipid metabolism in plants is much
less well understood than that in many other pathways in plant biology. In the
investigation reported here, we have characterized transcriptional regulatory
mechanisms controlling seed-specific FAD2 expression in sesame (Sesamum indicum).
FAD2 codes for extra-plastidial FAD2 desaturase, which catalyzes the conversion
of oleic acid to linoleic acid. Promoter analysis of the sesame FAD2 gene
(SeFAD2) using the beta-glucuronidase (GUS) reporter system demonstrated that the
- 660 to - 180 promoter region functions as a negative cis-element in the
seed-specific expression of the SeFAD2 gene. Sesame and Arabidopsis FAD2 genes
harbor one large intron within their 5'-untranslated region. These introns
conferred up to 100-fold enhancement of GUS expression in transgenic Arabidopsis
tissues as compared with intron-less controls. Prerequisite cis-elements for the
SeFAD2 intron-mediated enhancement of gene expression and the promoter-like
activity of SeFAD2 intron were identified. SeFAD2 transcripts were induced by
abscisic acid (ABA) in developing sesame seeds, and the - 660 to - 548 and - 179
to - 53 regions in the SeFAD2 promoter were implicated in ABA-responsive
signaling. Theses observations indicate that an intron-mediated regulatory
mechanism is involved in controlling not only the seed-specific expression of the
SeFAD2 gene but also the expression of plant FAD2 genes, which are essential for
the synthesis of polyunsaturated fatty acids.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16862401 [PubMed - indexed for MEDLINE]

1210: Obesity (Silver Spring). 2006 Jun;14(6):1003-9.

Increased leptin expression in the dorsal vagal complex suppresses adiposity
without affecting energy intake and metabolic hormones.

Boghossian S, Lecklin A, Dube MG, Kalra PS, Kalra SP.

Department of Neuroscience, University of Florida McKnight Brain Institute,
Gainesville, 32610-0244, USA.

OBJECTIVE: Increased leptin transgene expression locally in hypothalamic sites
suppresses weight and energy intake, enhances thermogenic energy expenditure, and
differentially modulates metabolic hormones for an extended period. We evaluated
whether a similar localized expression of leptin transgene in the dorsal vagal
complex (DVC) in the caudal brain stem that also displays the biologically
relevant leptin receptor would reproduce these varied responses and thus
demonstrate functional connectivity between the hypothalamus and DVC. RESEARCH
METHODS AND PROCEDURES: Adult female rats were microinjected with a recombinant
adeno-associated virus encoding either rat leptin or green fluorescent protein
gene (control) in the DVC. Food intake and body weight were monitored weekly, and
metabolic variables were analyzed at the end of 10 weeks. RESULTS AND DISCUSSION:
Increased leptin transgene expression in the DVC suppressed the time-related
increase in body weight accompanied by a transient decrease in food intake at
week 1 post-injection and little effect on thermogenic energy expenditure. That
suppression of weight was due to decreased adiposity is shown by the markedly
suppressed white adipose tissue-derived hormones, leptin and adiponectin.
Circulating concentrations of pancreatic insulin, gastric ghrelin, and glucose
levels were unchanged. This segregation of the varied effects of leptin
expression in hypothalamic sites vs. DVC endorses the view that among the various
endocrine organs under sympathetic nervous system control, only those
leptin-activated neural circuits in the hypothalamus that suppress weight and
adiposity on a long-term basis transverse through DVC en route to white adipose
tissue.

Publication Types:
Research Support, N.I.H., Extramural

PMID: 16861605 [PubMed - indexed for MEDLINE]

1211: Plant Physiol. 2006 Sep;142(1):54-62. Epub 2006 Jul 21.

Ectopic expression of KNOTTED1-like homeobox protein induces expression of
cytokinin biosynthesis genes in rice.

Sakamoto T, Sakakibara H, Kojima M, Yamamoto Y, Nagasaki H, Inukai Y, Sato Y,
Matsuoka M.

Field Production Science Center, Graduate School of Agricultural and Life
Sciences, The University of Tokyo, Nishi-Tokyo, Tokyo 188-0002, Japan.

Some phytohormones such as gibberellins (GAs) and cytokinins (CKs) are potential
targets of the KNOTTED1-like homeobox (KNOX) protein. To enhance our
understanding of KNOX protein function in plant development, we identified rice
(Oryza sativa) genes for adenosine phosphate isopentenyltransferase (IPT), which
catalyzes the rate-limiting step of CK biosynthesis. Molecular and biochemical
studies revealed that there are eight IPT genes, OsIPT1 to OsIPT8, in the rice
genome, including a pseudogene, OsIPT6. Overexpression of OsIPTs in transgenic
rice inhibited root development and promoted axillary bud growth, indicating that
OsIPTs are functional in vivo. Phenotypes of OsIPT overexpressers resembled those
of KNOX-overproducing transgenic rice, although OsIPT overexpressers did not form
roots or ectopic meristems, both of which are observed in KNOX overproducers.
Expression of two OsIPT genes, OsIPT2 and OsIPT3, was up-regulated in response to
the induction of KNOX protein function with similar kinetics to those of
down-regulation of GA 20-oxidase genes, target genes of KNOX proteins in dicots.
However, expression of these two OsIPT genes was not regulated in a feedback
manner. These results suggest that OsIPT2 and OsIPT3 have unique roles in the
developmental process, which is controlled by KNOX proteins, rather than in the
maintenance of bioactive CK levels in rice. On the basis of these findings, we
concluded that KNOX protein simultaneously decreases GA biosynthesis and
increases de novo CK biosynthesis through the induction of OsIPT2 and OsIPT3
expression, and the resulting high-CK and low-GA condition is required for
formation and maintenance of the meristem.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16861569 [PubMed - indexed for MEDLINE]

1212: J Biotechnol. 2006 Dec 15;127(1):161-6. Epub 2006 Jun 12.

Random amplified polymorphic DNA analysis of genetically modified organisms.

Yoke-Kqueen C, Radu S.

Department of Biomedical Science, Faculty of Medicine and Health Sciences,
Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
ykcheah@medic.upm.edu.my

Randomly amplified polymorphic DNA (RAPD) was used to analyzed 78 samples
comprises of certified reference materials (soya and maize powder), raw seeds
(soybean and maize), processed food and animal feed. Combination assay of two
arbitrary primers in the RAPD analysis enable to distinguish genetically modified
organism (GMO) reference materials from the samples tested. Dendrogram analysis
revealed 13 clusters at 45% similarity from the RAPD. RAPD analysis showed that
the maize and soybean samples were clustered differently besides the GMO and
non-GMO products.

PMID: 16860900 [PubMed - indexed for MEDLINE]

1213: Plant Cell Rep. 2006 Dec;25(12):1347-54. Epub 2006 Jul 21.

Erratum in:
Plant Cell Rep. 2006 Dec;25(12):1392.

Bisexual sterility conferred by the differential expression of barnase and
barstar: a simple and efficient method of transgene containment.

Kobayashi K, Munemura I, Hinata K, Yamamura S.

Iwate Biotechnology Research Center, 22-174-4 Narita, Kitakami, Iwate, 024-0003,
Japan. kappei@ibrc.or.jp

To establish a simple and an efficient system to minimize the environmental risk
of genetically modified plants, we tested the applicability of the
barnase/barstar system in conferring bisexual sterility; that is, in preventing
plants setting seeds by self-fertilization and out-crossing. Transgenic tobacco
plants were generated to express barnase, a cell death inducing ribonuclease,
under the control of the gamete-specific AtDMC1 promoter, and barstar, a specific
inhibitor of barnase, under the control of the ACT2 promoter, which is
constitutively active in almost all tissues except gametes. In contrast to
control plants harboring the barstar expression unit only, which set seeds
normally with self-pollination, all transformants harboring both barnase and
barstar were bisexually sterile. They produced aberrant anthers containing no
detectable pollen and failed to set seeds even after pollination with wild-type
tobacco pollen.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16858551 [PubMed - indexed for MEDLINE]

1214: Plant Cell Physiol. 2006 Sep;47(9):1195-205. Epub 2006 Jul 20.

Sl-ERF2, a tomato ethylene response factor involved in ethylene response and seed
germination.

Pirrello J, Jaimes-Miranda F, Sanchez-Ballesta MT, Tournier B, Khalil-Ahmad Q,
Regad F, Latchö© A, Pech JC, Bouzayen M.

UMR990 INRA/INP-ENSA Toulouse Gö©nomique et Biotechnologie des Fruits Avenue de
l'Agrobiopole, BP 32607, 31326 Castanet-Tolosan cedex, France.

Ethylene response factors (ERFs) are plant transcriptional regulators mediating
ethylene-dependent gene expression via binding to the GCC motif found in the
promoter region of ethylene-regulated genes. We report here on the structural and
functional characterization of the tomato Sl-ERF2 gene that belongs to a distinct
class of the large ERF gene family. Both spliced and unspliced versions of
Sl-ERF2 transcripts were amplified from RNA samples and the search in the public
tomato expressed sequence tag (EST) database confirmed the existence of the two
transcript species in a number of cDNA libraries. The unspliced transcript
contains two open reading frames yielding two hypothetical proteins, a small
highly truncated version lacking the APETALA2 domain and a bigger protein lacking
the N-terminal MCGGAAI(I)/(L) consensus peptide specific to ERF members from
subfamily IV. Nevertheless, functional Sl-ERF2 protein may only derive from
spliced transcripts since, depending on the tissue, the level of the spliced
transcript is much higher than that of the unspliced transcript. Sl-ERF2 is
expressed in all plant tissues tested, though its transcript accumulates
preferentially in germinating seeds and ripening fruit. Overexpression of the
Sl-ERF2 gene in transgenic tomato lines results in premature seed germination and
enhanced hook formation of dark-grown seedlings, which is indicative of increased
ethylene sensitivity. The expression of the mannanase2 gene is upregulated in
Sl-ERF2-overexpressing seeds, suggesting that Sl-ERF2 stimulates seed germination
through the induction of the mannanase2 gene. It is noteworthy that the
exaggerated hook phenotype is abolished when ethylene perception is blocked,
strongly suggesting that Sl-ERF2 requires other ethylene-dependent components to
impact the hook formation process.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16857696 [PubMed - indexed for MEDLINE]

1215: Plant J. 2006 Sep;47(5):675-86. Epub 2006 Jul 19.

Expression of ABA 8'-hydroxylases in relation to leaf water relations and seed
development in bean.

Yang SH, Zeevaart JA.

Department of Energy, Plant Research Laboratory, Michigan State University, East
Lansing, MI 48824-1312, USA.

In plants, the level of abscisic acid (ABA) is determined by synthesis and
catabolism. Hydroxylation of ABA at the 8' position is the key step in ABA
catabolism. This reaction is catalyzed by ABA 8'-hydroxylase, a cytochrome P450
(CYP). The cDNAs of PvCYP707A1 and PvCYP707A2 were isolated from bean (Phaseolus
vulgaris L.) axes treated with (+)-ABA and that of PvCYP707A3 from dehydrated
bean leaves. The recombinant PvCYP707A proteins expressed in yeast were
biochemically characterized. Yeast strains over-expressing any of the three
PvCYP707As were able to convert ABA to phaseic acid (PA). The microsomal
fractions from these yeast strains also exhibited ABA 8'-hydroxylase activity.
Expression of PvCYP707A3 in primary leaves was strongly increased by water
stress, whereas PvCYP707A1 and PvCYP707A2 mRNA levels were rapidly increased by
rehydration of water-stressed leaves. Northern blot analysis of PvCYP707As in
bean showed a high level of expression in the mature fruits, senescent leaves,
roots, seed coats and axes. All three PvCYP707As were expressed at varying
intensities throughout seed development. Imbibed seeds also had high PvCYP707A
mRNA levels. Thus, expression of PvCYP707As is both environmentally and
developmentally regulated. Transgenic Nicotiana sylvestris plants over-expressing
PvCYP707As displayed a wilty phenotype, and had reduced ABA levels and increased
PA levels. These results demonstrate that expression of PvCYP707As is the major
mechanism by which ABA catabolism is regulated in bean.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16856981 [PubMed - indexed for MEDLINE]

1216: Plant J. 2006 Sep;47(5):687-700. Epub 2006 Jul 19.

Visualization of PtdIns3P dynamics in living plant cells.

Vermeer JE, van Leeuwen W, Tobeö±a-Santamaria R, Laxalt AM, Jones DR, Divecha N,
Gadella TW Jr, Munnik T.

Section of Molecular Cytology, Swammerdam Institute for Life Sciences, University
of Amsterdam, Kruislaan 316, Amsterdam, The Netherlands.

To investigate PtdIns3P localization and function in plants, a fluorescent
PtdIns3P-specific biosensor (YFP-2xFYVE) was created. On lipid dot blots it bound
specifically and with high affinity to PtdIns3P. Transient expression in cowpea
protoplasts labelled vacuolar membranes and highly motile structures undergoing
fusion and fission. Stable expression in tobacco BY-2 cells labelled similar
motile structures, but labelled vacuolar membranes hardly at all. YFP-2xFYVE
fluorescence strongly co-localized with the pre-vacuolar marker AtRABF2b,
partially co-localized with the endosomal tracer FM4-64, but showed no overlap
with the Golgi marker STtmd-CFP. Treatment of cells with wortmannin, a PI3 kinase
inhibitor, caused the YFP-2xFYVE fluorescence to redistribute into the cytosol
and nucleus within 15 min. BY-2 cells expressing YFP-2xFYVE contained twice as
much PtdIns3P as YFP-transformed cells, but this had no effect on cell-growth or
stress-induced phospholipid signalling responses. Upon treatment with wortmannin,
PtdIns3P levels were reduced by approximately 40% within 15 min in both cell
lines. Stable expression of YFP-2xFYVE in Arabidopsis plants labelled different
subcellular structures in root compared with shoot tissues. In addition labelling
the motile structures common to all cells, YFP-2xFYVE strongly labelled the
vacuolar membrane in leaf epidermal and guard cells, suggesting that cell
differentiation alters the distribution of PtdIns3P. In dividing BY-2 cells,
YFP-2xFYVE-labelled vesicles surrounded the newly formed cell plate, suggesting a
role for PtdIns3P in cytokinesis. Together, these data show that YFP-2xFYVE may
be used as a biosensor to specifically visualize PtdIns3P in living plant cells.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16856980 [PubMed - indexed for MEDLINE]

1217: Genetics. 2006 Sep;174(1):317-29. Epub 2006 Jul 18.

Maternal gametophytic baseless1 is required for development of the central cell
and early endosperm patterning in maize (Zea mays).

Gutiö©rrez-Marcos JF, Costa LM, Evans MM.

Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, United
Kingdom.

In angiosperms, double fertilization of an egg cell and a central cell with two
sperm cells results in the formation of a seed containing a diploid embryo and a
triploid endosperm. The extent to which the embryo sac controls postfertilization
events in the seed is unknown. The novel gametophytic maternal-effect maize
mutation, baseless1 (bsl1) affects central cell development within the embryo
sac, frequently by altering the position of the two polar nuclei. Despite this
irregularity, fertilization is as efficient as in wild type. The spatial
expression of basal endosperm-specific transcripts is altered in free-nuclear and
cellular mutant endosperms. At later stages of seed development, bsl1
predominantly affects development of the basal endosperm transfer layer (BETL).
When bsl1/+ diploid plants were pollinated by wild-type tetraploid plants, the
BETL abnormalities observed in bsl1/bsl1/+/+ tetraploid endosperms were diverse
and of variable severity. Moreover, the frequency of kernels with severely
perturbed BETL development correlated with the percentage of severely affected
bsl1 central cells. Therefore, BSL1 is likely required in the central cell before
fertilization for correct BETL patterning to occur. These findings provide new
genetic evidence that a maternal gametophytic component is necessary for correct
endosperm patterning.

PMID: 16849604 [PubMed - indexed for MEDLINE]

1218: Genetics. 2006 Sep;174(1):309-16. Epub 2006 Jul 18.

Effects of sex and insulin/insulin-like growth factor-1 signaling on performance
in an associative learning paradigm in Caenorhabditis elegans.

Vellai T, McCulloch D, Gems D, Kovö¡cs AL.

Department of Genetics, Eö¶tvö¶s Lorö¡nd University, Budapest, H-1117, Hungary.
vellai@falco.elte.hu

Learning is an adaptive change in behavior in response to environmental stimuli.
In mammals, there is a distinct female bias to learn skills that is still
unprecedented in other animal taxa. Here we have investigated the biological
determinants of performance in an associative learning paradigm in the nematode
Caenorhabditis elegans. Using an assay of chemotactic reactions associated with
food deprivation, wild-type male worms show inferior learning ability relative to
hermaphrodites. Sex-based learning difference is therefore an ancient
evolutionary feature appearing even in relatively simple animals. C. elegans
mutants with reduced insulin/IGF-1 signaling also exhibit a greatly reduced
learning ability in this assay. In addition, hyperactivation of insulin/IGF-1
signaling through loss-of-function mutations in the PTEN phosphatase daf-18, a
negative regulator of insulin/IGF-1 signaling, enhances learning ability beyond
that of wild type. According to our epistasis analysis, the effect of DAF-2 on
learning acts via phosphatidylinositol 3,4,5-trisphosphate (PIP(3)) production,
but not the DAF-16 FOXO transcription factor. This implies that the signaling
pathway from DAF-2 affecting this learning paradigm branches between PIP(3)
production and DAF-16. However, learning capacity of nematodes is lowered by
loss-of-function mutations in daf-16, suggesting involvement of noninsulin/IGF-1
signaling-dependent DAF-16 activation in learning. Potentially, sex and
insulin/IGF-1 signaling affect performance in this learning assay via effects on
the neurobiology of learning.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16849598 [PubMed - indexed for MEDLINE]

1219: Ann Bot (Lond). 2006 Oct;98(4):819-25. Epub 2006 Jul 18.

Evaluation of metabolic alteration in transgenic rice overexpressing
dihydroflavonol-4-reductase.

Takahashi H, Hayashi M, Goto F, Sato S, Soga T, Nishioka T, Tomita M,
Kawai-Yamada M, Uchimiya H.

Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1
Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan.

BACKGROUND AND AIMS: Previous studies have shown that transgenic rice plants
overexpressing YK1, which possesses dihydroflavonol-4-reductase (DFR) activity,
showed biotic and abiotic stress tolerance. High throughput profiles of
metabolites have also been shown in such transgenic plants by Fourier transform
ion cyclotron mass spectrometry. In this study, capillary electrophoresis mass
spectrometry analysis (CE/MS) was employed to identify precise metabolites such
as organic acids, amino acids and sugars. METHODS: Using CE/MS, we analysed
several metabolites of glycolysis, the tricarboxylic acid (TCA) cycle and the
pentose phosphate pathway. In addition, the concentrations of sugars and ion were
quantified. KEY RESULTS: In YK1 (DFR)-overexpressing plants, the concentrations
of cis-aconitate, isocitrate and 2-oxoglutarate were higher in leaves, whereas
those of fructose-1,6-bisphosphate and glyceraldehyde-3-phosphate were lower in
roots. In seeds, the amounts of free amino acids and metals were altered, whereas
sugars in seeds were kept constant. In YK1 calli, an approx. 3-fold increase in
glutathione was observed, whereas the activities of glutathione peroxidase and
glutathione reductase were concomitantly increased. CONCLUSIONS: The
overexpression of YK1 (DFR) was associated with slight changes in the amounts of
several metabolites analysed in whole plants, whilst glutathione derivatives were
substantially increased in suspension-cultured cells.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16849376 [PubMed - indexed for MEDLINE]

1220: Plant Cell Rep. 2006 Dec;25(12):1336-46. Epub 2006 Jul 18.

Targeted expression of redesigned and codon optimised synthetic gene leads to
recrystallisation inhibition and reduced electrolyte leakage in spring wheat at
sub-zero temperatures.

Khanna HK, Daggard GE.

Centre for Systems Biology, Faculty of Sciences, University of Southern
Queensland, Toowoomba, Qld., 4350, Australia. h.khanna@qut.edu.au

Antifreeze proteins (AFPs) adsorb to ice crystals and inhibit their growth,
leading to non-colligative freezing point depression. Crops like spring wheat,
that are highly susceptible to frost damage, can potentially be made frost
tolerant by expressing AFPs in the cytoplasm and apoplast where ice
recrystallisation leads to cellular damage. The protein sequence for HPLC-6
alpha-helical antifreeze protein from winter flounder was rationally redesigned
after removing the prosequences in the native protein. Wheat nuclear gene
preferred amino acid codons were used to synthesize a recombinant antifreeze
gene, rAFPI. Antifreeze protein was targeted to the apoplast using a Murine
leader peptide sequence from the mAb24 light chain or retained in the endoplasmic
reticulum using C-terminus KDEL sequence. The coding sequences were placed
downstream of the rice Actin promoter and Actin-1 intron and upstream of the
nopaline synthase terminator in the plant expression vectors. Transgenic wheat
lines were generated through micro projectile bombardment of immature embryos of
spring wheat cultivar Seri 82. Levels of antifreeze protein in the transgenic
lines without any targeting peptide were low (0.06-0.07%). The apoplast-targeted
protein reached a level of 1.61% of total soluble protein, 90% of which was
present in the apoplast. ER-retained protein accumulated in the cells at levels
up to 0.65% of total soluble proteins. Transgenic wheat line T-8 with
apoplast-targeted antifreeze protein exhibited the highest levels of antifreeze
activity and provided significant freezing protection even at temperatures as low
as -7 degrees C.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16847628 [PubMed - indexed for MEDLINE]

1221: Planta. 2006 Dec;225(1):89-102. Epub 2006 Jul 15.

CHRD, a plant member of the evolutionarily conserved YjgF family, influences
photosynthesis and chromoplastogenesis.

Leitner-Dagan Y, Ovadis M, Zuker A, Shklarman E, Ohad I, Tzfira T, Vainstein A.

The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture,
Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew
University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel.

Studies on the carotenoid-overaccumulating structures in chromoplasts have led to
the characterization of proteins termed plastid lipid-associated proteins (PAPs),
involved in the sequestration of hydrophobic compounds. Here we characterize the
PAP CHRD, which, based on sequence homology, belongs to a highly conserved group
of proteins, YER057c/YjgF/UK114, involved in the regulation of basic and vital
cellular processes in bacteria, yeast and animals. Two nuclear genes were
characterized in tomato plants: one (LeChrDc) is constitutively expressed in
various tissues and the other (LeChrDi) is induced by stress in leaves and is
upregulated by developmental cues in floral tissues. Using RNAi and antisense
approaches, we show their involvement in biologically significant processes such
as photosynthesis. The quantum yield of photosynthetic electron flow in
transgenic tomato leaves with suppressed LeChrDi/c expression was 30-50% of their
control, non-transgenic counterparts and was ascribed to lower PSI activity.
Transgenic flowers with suppressed LeChrDi/c also accumulated up to 30% less
carotenoids per unit protein as compared to control plants, indicating an
interrelationship between PAPs and floral-specific carotenoid accumulation in
chromoplasts. We suggest that CHRD's role in the angiosperm reproductive unit may
be a rather recent evolutionary development; its original function may have been
to protect the plant under stress conditions by preserving plastid functionality.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16845531 [PubMed - indexed for MEDLINE]

1222: Planta. 2007 Jan;225(2):287-300. Epub 2006 Jul 15.

The promoter of the wheat puroindoline-a gene (PinA) exhibits a more complex
pattern of activity than that of the PinB gene and is induced by wounding and
pathogen attack in rice.

Evrard A, Meynard D, Guiderdoni E, Joudrier P, Gautier MF.

INRA, UMR1096 PIA, 2 place Viala, 34060, Montpellier Cedex 01, France.

Puroindolines form the molecular basis of wheat grain hardness. However, little
is known about puroindoline gene regulation. We previously reported that the
Triticum aestivum puroindoline-b gene (PinB) promoter directs beta-glucuronidase
gene (uidA) seed-specific expression in transgenic rice. In this study, we
isolated a puroindoline-a gene (PinA), analyzed PinA promoter activity by 5'
deletions and compared PinA and PinB promoters in transgenic rice. Seeds of
PinA-1214 and PinB-1063 transgenic plants strongly expressed uidA in endosperm,
in the aleurone layer and in epidermis cells in a developmentally regulated
manner. The GUS activity was also observed in PinA-1214 embryos. Whereas the PinB
promoter is seed specific, the PinA promoter also directed, but to a lower level,
uidA expression in roots of seedlings and in the vascular tissues of palea and
pollen grains of dehiscent anthers during flower development. In addition, the
PinA promoter was induced by wounding and by Magnaporthe grisea. By deletion
analysis, we showed that the "390-bp" PinA promoter drives the same expression
pattern as the "1214-bp" promoter. Moreover, the "214-bp" PinA promoter drives
uidA expression solely in pollen grains of dehiscent anthers. The presence of
putative cis-regulatory elements that may be related to PinA expression is
discussed from an evolutionary point of view. By electrophoretic mobility shift
assay, we showed that putative cis-elements (WUN-box, TCA motifs and as-1-like
binding sites) whose presence in the PinA promoter may be related to wounding
and/or the pathogen response form complexes with nuclear extracts isolated from
wounded wheat leaves.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16845527 [PubMed - indexed for MEDLINE]

1223: Planta. 2007 Jan;225(2):403-11. Epub 2006 Jul 15.

Enhanced Cd2+ -selective root-tonoplast-transport in tobaccos expressing
Arabidopsis cation exchangers.

Koren'kov V, Park S, Cheng NH, Sreevidya C, Lachmansingh J, Morris J, Hirschi K,
Wagner GJ.

Department of Plant and Soil Sciences, Plant Biology Program, University of
Kentucky, Lexington, KY 40546, USA.

Several Arabidopsis CAtion eXchangers (CAXs) encode tonoplast-localized
transporters that appear to be major contributors to vacuolar
accumulation/sequestration of cadmium (Cd(2+)), an undesirable pollutant ion that
occurs in man largely as a result of dietary consumption of aerial tissues of
food plants. But, ion-selectivity of individual CAX transporter types remains
largely unknown. Here, we transformed Nicotiana tabacum with several CAX genes
driven by the Cauliflower Mosaic Virus (CaMV) 35S promoter and monitored divalent
cation transport in root-tonoplast vesicles from these plants in order to select
particular CAX genes directing high Cd(2+) antiporter activity in root tonoplast.
Comparison of seven different CAX genes indicated that all transported Cd(2+),
Ca(2+), Zn(2+), and Mn(2+) to varying degrees, but that CAX4 and CAX2 had high
Cd(2+) transport and selectivity in tonoplast vesicles. CAX4 driven by the CaMV
35S and FS3 [figwort mosaic virus (FMV)] promoters increased the magnitude and
initial rate of Cd(2+)/H(+) exchange in root-tonoplast vesicles. Ion selectivity
of transport in root-tonoplast vesicles isolated from FS3::CAX4-expressing plant
lines having a range of gene expression was Cd(2+)>Zn(2+)>>Ca(2+)>>Mn(2+) and the
ratios of maximal Cd(2+) (and Zn(2+)) versus maximal Ca(2+) and Mn(2+) transport
were correlated with the levels of CAX4 expression. Root Cd accumulation in high
CAX4 and CAX2 expressing lines was increased in seedlings grown with 0.02 muM Cd.
These observations are consistent with a model in which expression of an
Arabidopsis-gene-encoded, Cd(2+)-efficient antiporter in host plant roots results
in greater root vacuole Cd(2+) transport activity, increased root Cd
accumulation, and a shift in overall root tonoplast ion transport selectivity
towards higher Cd(2+) selectivity. Results support a model in which certain CAX
antiporters are somewhat more selective for particular divalent cations.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

PMID: 16845524 [PubMed - indexed for MEDLINE]

1224: Biochim Biophys Acta. 2006 Sep;1760(9):1434-44. Epub 2006 May 24.

Biochemical limitations to high-level expression of humanized monoclonal
antibodies in transgenic maize seed endosperm.

Law RD, Russell DA, Thompson LC, Schroeder SC, Middle CM, Tremaine MT, Jury TP,
Delannay X, Slater SC.

Monsanto Protein Technologies, 8520 University Green, Middleton, WI 53562, USA.
david.law@lakeheadu.ca

Transgenic plants are potentially valuable systems for the large scale
manufacture of therapeutic proteins. To improve this technology, determining the
importance of transgene transcript levels on protein accumulation in sink tissues
during their development is crucial. In transgenic maize (Zea mays L.) plants
expressing humanized monoclonal antibodies (mAbs) in their seed endosperm,
steady-state kappa light chain (LC) and gamma heavy chain (HC) mRNA levels were
quantified during development and compared to the levels of fully-assembled mAb
protein present at seed maturity. RNA blots and non-reducing SDS-PAGE western
immunoblots revealed that steady-state LC and HC mRNA and protein levels were
undetectable at 10 days after pollination (DAP) but increased quickly thereafter
in three transgenic events expressing different mAb molecules. Similar to
gamma-zein mRNA, LC and HC messages were highly abundant between 15 and 25 DAP.
Quantitative RNA blots and western immunoblots showed that steady-state LC
transcript levels during development correlated extremely closely with protein
levels in mature seed (r(2)=0.99). For HC, this correlation was not as strong
(r(2)=0.85). Consistent with this finding, concomitantly increasing the zygosity
levels of the LC and HC transgenes enhanced mAb concentration in mature seed, in
contrast to increasing the copy number of the transgene insert, which did not
correlate with high seed mAb levels. The results indicate that high-level
expression of fully-assembled mAb protein in maize endosperm was favored by high
LC and HC mRNA levels and was largely limited by HC protein concentration.

PMID: 16842925 [PubMed - indexed for MEDLINE]

1225: Mol Ecol. 2006 Aug;15(9):2677-85.

Impact of Bt maize pollen (MON810) on lepidopteran larvae living on accompanying
weeds.

Gathmann A, Wirooks L, Hothorn LA, Bartsch D, Schuphan I.

Aachen University, Institute of Environmental Research, Chair of Ecology,
Ecotoxicology and Ecochemistry, Worringerweg 1, 52062 Aachen, Germany.
achim.gathmann@bvl.bund.de

Environmental risks of Bt maize, particularly pollen drift from Bt maize, were
assessed for nontarget lepidopteran larvae in maize field margins. In our
experimental approach, we carried out 3-year field trials on 6 ha total. Three
treatments were used in a randomized block design with eight replications
resulting in 24 plots: (i) near-isogenic control variety without insecticide
(control), (ii) near-isogenic control variety with chemical insecticide
(Baytroid) and (iii) Bt maize expressing the recombinant toxin. We established a
weed strip (20 x 1 m) in every plot consisting of a Chenopodium album
(goosefoot)/Sinapis alba (mustard) mixture. In these strips we measured diversity
and abundance of lepidopteran larvae during maize bloom and pollen shed. C. album
hosted five species but all in very low densities; therefore data were not
suitable for statistical analysis. S. alba hosted nine species in total. Most
abundant were Plutella xylostella and Pieris rapae. For these species no
differences were detected between the Bt treatment and the control, but the
chemical insecticide treatment reduced larval abundance significantly.
Conclusions regarding experimental methodology and results are discussed in
regard to environmental risk assessment and monitoring of genetically modified
organisms.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 16842436 [PubMed - indexed for MEDLINE]

1226: Plant Cell Rep. 2006 Dec;25(12):1380-6. Epub 2006 Jul 14.

Enhanced tolerance of transgenic potato plants expressing both superoxide
dismutase and ascorbate peroxidase in chloroplasts against oxidative stress and
high temperature.

Tang L, Kwon SY, Kim SH, Kim JS, Choi JS, Cho KY, Sung CK, Kwak SS, Lee HS.

Environmental Biotechnology Research Center, Korea Research Institute of
Bioscience and Biotechnology, Yuseong, Daejeon, 305-806, Korea.

Oxidative stress is a major damaging factor for plants exposed to environmental
stresses. In order to develop transgenic potato plants with enhanced tolerance to
environmental stress, the genes of both Cu/Zn superoxide dismutase and ascorbate
peroxidase were expressed in chloroplasts under the control of an oxidative
stress-inducible SWPA2 promoter (referred to as SSA plants). SSA plants showed
enhanced tolerance to 250 microM methyl viologen, and visible damage in SSA
plants was one-fourth that of non-transgenic (NT) plants that were almost
destroyed. In addition, when SSA plants were treated with a high temperature of
42 degrees C for 20 h, the photosynthetic activity of SSA plants decreased by
only 6%, whereas that of NT plants decreased by 29%. These results suggest that
the manipulation of the antioxidative mechanism of the chloroplasts may be
applied in the development of industrial transgenic crop plants with increased
tolerance to multiple environmental stresses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16841217 [PubMed - indexed for MEDLINE]

1227: Plant Cell Rep. 2006 Dec;25(12):1355-61. Epub 2006 Jul 14.

A red fluorescent protein, DsRed2, as a visual reporter for transient expression
and stable transformation in soybean.

Nishizawa K, Kita Y, Kitayama M, Ishimoto M.

National Agricultural Research Center for Hokkaido Region, Toyohira, Sapporo,
Hokkaido, 062-8555, Japan.

Fluorescent proteins such as green fluorescent protein (GFP) from Aequorea
victoria are often used as markers for transient expression and stable
transformation in plants, given that their detection does not require a substrate
and they can be monitored in a nondestructive manner. We have now evaluated the
red fluorescent protein DsRed2 (a mutant form of DsRed from Discosoma sp.) for
its suitability as a visual marker in combination with antibiotic selection for
genetic transformation of soybean [Glycine max (L.) Merrill]. Transient and
stable expression of DsRed2 in somatic embryos was readily detected by
fluorescence microscopy, allowing easy confirmation of gene introduction. We
obtained several fertile transgenic lines, including homozygous lines, that grew
and produced seeds in an apparently normal manner. The red fluorescence of DsRed2
was detected by fluorescence microscopy without background fluorescence in both
leaves and seeds of the transgenic plants. Furthermore, in contrast to seeds
expressing GFP, those expressing DsRed2 were readily identifiable even under
white light by the color conferred by the transgene product. The protein
composition of seeds was not affected by the introduction of DsRed2, with the
exception of the accumulation of DsRed2 itself, which was detectable as an
additional band on electrophoresis. These results indicate that DsRed2 is a
suitable reporter (even more suitable than GFP) for genetic transformation of
soybean.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16841215 [PubMed - indexed for MEDLINE]

1228: S Afr Med J. 2006 Jun;96(6):509-10.

Genetically modified crops--playing a positive role in sustainable development in
Africa.

Thomson JA.

Department of Molecular and Cell Biology, University of Cape Town, South Africa.
jat@science.uct.ac.za

PMID: 16841131 [PubMed - indexed for MEDLINE]

1229: Nat Biotechnol. 2006 Jul;24(7):753.

Comment in:
Nat Biotechnol. 2006 Nov;24(11):1327-9; author reply 1331-3. Nat Biotechnol. 2006 Nov;24(11):1329-31; author reply 1331-3. Nat Biotechnol. 2006 Nov;24(11):1329; author reply 1331-3.

Do cisgenic plants warrant less stringent oversight?

Schouten HJ, Krens FA, Jacobsen E.

Publication Types:
Letter

PMID: 16841052 [PubMed - indexed for MEDLINE]

1230: Nat Biotechnol. 2006 Jul;24(7):748-9.

Regulatory slowdown on GM crop decisions.

Jaffe G.

Publication Types:
Letter

PMID: 16841049 [PubMed - indexed for MEDLINE]

1231: Nat Biotechnol. 2006 Jul;24(7):735.

Elliot Entis.

Powell K.

Publication Types:
News

PMID: 16841044 [PubMed - indexed for MEDLINE]

1232: Biotechnol Bioeng. 2006 Oct 20;95(3):526-32.

Restraining Erwinia virulence by expression of N-acyl homoserine lactonase gene
pro3A-aiiA in Bacillus thuringiensis subsp leesis.

Zhu C, Yu Z, Sun M.

State Key Laboratory of Agricultural Microbiology, College of Life Science and
Technology, Huazhong Agricultural University, Wuhan 430070, China.

To widen the biological control function of a genetically modified Bacillus
thuringiensis subsp leesis strain BMB-005, an acyl homoserine lactonase (AHL
lactonase) gene aiiA transcribed by the promoter of insecticidal crystal protein
coding gene cry3A, was transformed into strain BMB-005. The amount of AHL
lactonase protein produced by transformant BMB821A was 2.4-fold more than that
produced by BMB-005. AHL-degradation assay showed that transformant BMB821A could
degrade more AHLs molecules than the original strain BMB-005. The result of
Erwinia carotovora pathogenicity test showed that the parental strain BMB-005 had
no restraint of Erwinia infection, but the transformants exhibited strong
restraint of E. carotovora infection on potato slices and cactus stems.
Insecticidal bioassay against lepidopteran Spodoptera exigua showed that both
strain BMB-005 and transformant BMB821A were toxic to S. exigua. The toxicity of
transformant BMB821A (LC(50) was 3.8) was a little attenuated comparing with the
toxicity of the original strain BMB-005 (LC(50) was 2.9). The B. thuringiensis
strain BMB-005 has high toxicity against Helicoverpa armigera, Plutella
xylostella, and S. exigua. This work provided new strategy for developing
genetically engineered multi-functional B. thuringiensis strain that possesses
insecticidal activity together with restraint of bacterial pathogenicity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16838380 [PubMed - indexed for MEDLINE]

1233: J Exp Bot. 2006;57(11):2887-97. Epub 2006 Jul 12.

Divergence in spatial expression patterns and in response to stimuli of
tandem-repeat paralogues encoding a novel class of proline-rich proteins in Oryza
sativa.

Wang R, Chong K, Wang T.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan,
Haidianqu, Beijing 100093, China.

Gene duplication has been recognized as a major route to supply raw sequences for
genome novelty in evolution, but the mechanism underlying the retention of
duplicate genes is not fully understood yet. Divergence in spatial expression
patterns was investigated here and in response to stimuli of the four members of
a rice proline-rich protein gene family (OsPRP1), which encode a class of
proline-rich proteins. The four paralogues are tandemly organized within a 20 kbp
range of chromosome 10 without any interval of other open reading frames and with
a median K(S) value of 0.474. These paralogues showed little similarity in their
regulatory regions but high conservation in coding regions. Search of an
intergenomic cis-element database predicted their promoter regions with divergent
cis-element fingerprints. Further expression analyses involving different
tissues/organs and nine types of stimuli by a promoter::GUS-fusion strategy
revealed that the four paralogues were expressed mainly in vascular cylinders of
different organs and showed diversity in tissue/organ specificity and in response
to these stimuli, with some overlapping expression. Furthermore, these data show
that OsPRP1.2 appeared to inherit most of the functions from their
multifunctional progenitor, whereas the other three genes diverged after
duplication events. Thus, the retention of paralogues in a multigene family seems
to require a more complicated diversification process than originally thought. In
addition, the promoter::GUS strategy is a powerful way to explore function
divergence of a tandem-repeat gene family.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16837535 [PubMed - indexed for MEDLINE]

1234: Risk Anal. 2006 Jun;26(3):845-58.

Genetically engineered plants, endangered species, and risk: a temporal and
spatial exposure assessment for Karner blue butterfly larvae and Bt maize pollen.

Peterson RK, Meyer SJ, Wolf AT, Wolt JD, Davis PM.

Agricultural and Biological Risk Assessment, Dept. of Land Resources and
Environmental Sciences, 334 Leon Johnson Hall, Montana State University, Bozeman,
MT 59717-3120, USA. bpeterson@montana.edu

Genetically engineered maize (Zea mays) containing insecticidal endotoxin
proteins from Bacillus thuringiensis (Bt) delta-endotoxin proteins has been
adopted widely in the Midwestern United States. The proteins are toxic to several
lepidopteran species and because a variety of maize tissues, including pollen,
may express the endotoxins, the probability of exposure to nontarget species,
including endangered species, needs to be understood. The objective of this study
was to assess the potential temporal and spatial exposure of endangered Karner
blue butterfly larvae (Lycaeides melissa samuelis) to Bt maize pollen in
Wisconsin using probabilistic exposure techniques and geographic information
systems analysis. Based on degree-day modeling of butterfly phenology and maize
pollen shed, there is some potential for temporal exposure of larvae to maize
pollen. However, in the majority of years and locations, maize pollen shed most
likely will occur after the majority of larval feeding on wild lupine (Lupinus
perennis). The spatial analysis indicates that some Karner blue butterfly
populations occur in close proximity to maize fields, but in the vast majority of
cases the butterfly's host plant and maize fields are separated by more than 500
m. A small number of potential or existing Karner blue butterfly sites are
located near maize fields, including sites in two of the four counties where
temporal overlap is most likely. The exposure assessment indicates that these two
counties should receive the highest priority to determine if Karner blue
butterfly larvae are actually at risk and then, if needed, to reduce or prevent
exposure.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16834638 [PubMed - indexed for MEDLINE]

1235: Risk Anal. 2006 Jun;26(3):657-70.

Examining consumer behavior toward genetically modified (GM) food in Britain.

Spence A, Townsend E.

RASPH, School of Psychology, University of Nottingham, Nottingham, UK.
lpxas@psychology.nottingham.ac.uk

This study examined behavior toward genetically modified (GM) food in a British
community-based sample. We used an equivalent gain task in which participants
actually received the options they chose to encourage truthful responding. In
conjunction with this, theory of planned behavior (TPB) components were evaluated
so as to examine the relative importance of behavioral influences in this domain.
Here, the TPB was extended to include additional components to measure
self-identity, moral norms, and emotional involvement. Results indicated that the
monetary amounts participants accepted in preference to GM food were
significantly lower than those accepted in preference to non-GM food. However,
the vast majority of participants were indifferent between GM and non-GM food
options. All TPB components significantly predicted behavioral intentions to try
GM food, with attitudes toward GM being the strongest predictor. Self-identity
and emotional involvement were also found to be significant predictors of
behavioral intentions but moral norms were not. In addition, behavioral
intentions significantly predicted behavior; however, PBC did not. An additional
measure of participants' propensity to respond in a socially desirable manner
indicated that our results were not influenced by self-presentation issues,
giving confidence to our findings. Overall, it appears that the majority of
participants (74.5%) would purchase GM food at some price.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16834625 [PubMed - indexed for MEDLINE]

1236: J Exp Bot. 2006;57(11):2613-25. Epub 2006 Jul 10.

A metabonomic study of transgenic maize (Zea mays) seeds revealed variations in
osmolytes and branched amino acids.

Manetti C, Bianchetti C, Casciani L, Castro C, Di Cocco ME, Miccheli A, Motto M,
Conti F.

Dipartimento di Chimica, Universitö  degli studi di Roma La Sapienza, Piazzale
Aldo Moro 5, I-00185 Roma, Italy and Istituto Sperimentale per la Cerealicoltura,
Sez. Bergamo, Via Stezzano 24, I-24126 Bergamo, Italy. c.manetti@caspur.it

The aim of the research was to investigate metabolic variations associated with
genetic modifications in the grains of Zea mays using metabonomic techniques.
With this in mind, the non-targeted characteristic of the technique is useful to
identify metabolites peculiar to the genetic modification and initially
undefined. The results obtained showed that the genetic modification, introducing
Cry1Ab gene expression, induces metabolic variations involving the primary
nitrogen pathway. Concerning the methodological aspects, the experimental
protocol used has been applied in this field for the first time. It consists of a
combination of partial least square-discriminant analysis and principal component
analysis. The most important metabolites for discrimination were selected and the
metabolic correlations linking them are identified. Principal component analysis
on selected signals confirms metabolic variations, highlighting important details
about the changes induced on the metabolic network by the presence of a Bt
transgene in the maize genome.

PMID: 16831843 [PubMed - indexed for MEDLINE]

1237: Plant Mol Biol. 2006 Jun;61(3):525-35.

High lysine and high tryptophan transgenic maize resulting from the reduction of
both 19- and 22-kD alpha-zeins.

Huang S, Frizzi A, Florida CA, Kruger DE, Luethy MH.

Mystic Research, Monsanto Company, 62 Maritime Drive, Mystic, CT 06355, USA.
shihshieh.huang@monsanto.com

The major maize seed storage proteins, zeins, are deficient in lysine and
tryptophan content, which contribute to the poor nutritional quality of corn.
Whether through the identification of mutations or genetic engineering, kernels
with reduced levels of zein proteins have been shown to have increased levels of
lysine and tryptophan. It has been hypothesized that these increases are due to
the reduction of lysine-poor zeins and a pleiotropic increase in the lysine-rich
non-zein proteins. By transforming maize with constructs expressing chimeric
double-stranded RNA, kernels derived from stable transgenic plants displayed
significant declines in the accumulation of both 19- and 22-kD alpha-zeins, which
resulted in higher lysine and tryptophan content than previously reported for
kernels with reduced zein levels. The observation that lysine and tryptophan
content is correlated with the protein levels measured in transgenic maize
kernels is consistent with the hypothesis that a pleiotropic increase in non-zein
proteins is contributing to an improved amino acid balance. In addition, a large
increase in accumulation of free amino acids, consisting predominantly of
asparagine, aspartate and glutamate, was observed in the zein reduction kernels.

Publication Types:
Evaluation Studies

PMID: 16830184 [PubMed - indexed for MEDLINE]

1238: Plant Mol Biol. 2006 Jun;61(3):469-89.

A homeodomain leucine zipper gene from Craterostigma plantagineum regulates
abscisic acid responsive gene expression and physiological responses.

Deng X, Phillips J, Brö¤utigam A, Engströ¶m P, Johannesson H, Ouwerkerk PB, Ruberti
I, Salinas J, Vera P, Iannacone R, Meijer AH, Bartels D.

Max-Planck-Institut fö¼r Zö¼chtungsforschung, Carl-von-Linnö©-Weg 10, Cologne
D-50829, Germany.

A subset of homeodomain leucine zipper proteins (HDZip) play a role in regulating
adaptation responses including developmental adjustment to environmental cues in
plants. Here we report the structural and functional characterisation of a
dehydration responsive nuclear-targeted HDZip transcriptional regulator, CpHB-7.
DNA-protein interaction studies suggest that CDeT6-19, a known ABA and
dehydration responsive dehydrin gene, is a potential target gene of CpHB-7 in the
desiccation-tolerant plant Craterostigma plantagineum. Transgenic plants that
ectopically express CpHB-7 display reduced sensitivity towards ABA during seed
germination and stomatal closure. Expression analysis reveals that genes with
induced or repressed expression in CpHB-7 ectopic expression lines are either
mostly repressed or induced by ABA, drought or salt treatment respectively, thus
demonstrating that CpHB-7 modifies ABA-responsive gene expression as a negative
regulator. CpHB-7 gene expression is also linked to early organ development,
leading to the suggestion that CpHB-7 is functionally similar to the Arabidopsis
transcription factor, ATHB-6.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16830180 [PubMed - indexed for MEDLINE]

1239: Plant Mol Biol. 2006 Jun;61(3):415-30.

Silencing of an anther-specific zinc-finger gene, MEZ1, causes aberrant meiosis
and pollen abortion in petunia.

Kapoor S, Takatsuji H.

Developmental Biology Laboratory, Plant Physiology Department, National Institute
of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba 305-8602, Ibaraki, Japan.

MEZ1 (MEiosis-associated Zinc-finger protein 1) was first isolated as an
anther-specific cDNA from Petunia hybrida. In the present study, we report its
functional characterization, including its spatial and temporal expression
profiles and phenotypes in MEZ1-silenced plants. MEZ1 transcripts were
specifically localized in pollen mother cells during early stages of anther
development, and were later distributed in vegetative tissues in anthers.
Silencing of MEZ1 by cosuppression resulted in several anomalies during male
meiosis that included inability of chromosomes to condense, loss of meiotic
synchrony, and premature and apparently uncontrolled cytokinetic events.
Consequently, by the end of meiosis 8-10 cells, instead of the normal 4, with
varying DNA contents were formed in the MEZ1-silenced meiocytes. Most of these
aborted prematurely, and those that matured had a distinctive morphology.
MEZ1-silenced plants were female sterile when pollinated with wild-type pollen
but they infrequently produced a few seeds upon self-pollination. Resulting T1
plants had increased ploidy levels and exhibited severe anomalies during male
meiosis, rendering them completely sterile. We discuss possible role of MEZ1 in
the proper progression of plant meiosis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16830177 [PubMed - indexed for MEDLINE]

1240: Plant Cell. 2006 Aug;18(8):1873-86. Epub 2006 Jul 7.

AUXIN RESPONSE FACTOR8 is a negative regulator of fruit initiation in
Arabidopsis.

Goetz M, Vivian-Smith A, Johnson SD, Koltunow AM.

Commonwealth Scientific and Industrial Research Organization, Division of Plant
Industry, Horticulture Unit, Glen Osmond, SA 5064, Australia.

Fruit and seed formation in plants is normally initiated after pollination and
fertilization, and, in the absence of fertilization, flowers senesce. In the
Arabidopsis thaliana mutant fruit without fertilization, a mutation in AUXIN
RESPONSE FACTOR8 (ARF8) results in the uncoupling of fruit development from
pollination and fertilization and gives rise to seedless (parthenocarpic) fruit.
Parthenocarpy was confirmed in two additional recessive alleles and was caused by
mutations within the coding region of ARF8. Genetic experiments indicate that
ARF8 acts as an inhibitor to stop further carpel development in the absence of
fertilization and the generation of signals required to initiate fruit and seed
development. Expression of ARF8 was found to be regulated at multiple levels, and
transcriptional autoregulation of ARF8 was observed. Analysis of plants
transformed with a transcriptional P(ARF8):beta-glucuronidase (GUS) construct or
a translational ARF8:GUS fusion construct displayed distinct developmental
regulation of the reporter in floral tissues involved in pollination and
fertilization and in the carpel wall. After fertilization, the level of GUS
activity declined in the developing seed, while in unfertilized ovules that are
destined to senesce, ARF8:GUS expression spread throughout the ovule. This is
consistent with a proposed role for ARF8 in restricting signal transduction
processes in ovules and growth in pistils until the fruit initiation cue.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16829592 [PubMed - indexed for MEDLINE]

1241: Plant J. 2006 Aug;47(4):519-31. Epub 2006 Jul 10.

A brassinolide-suppressed rice MADS-box transcription factor, OsMDP1, has a
negative regulatory role in BR signaling.

Duan K, Li L, Hu P, Xu SP, Xu ZH, Xue HW.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun Road, 100093
Beijing, China.

The MADS-box transcription factor-encoding genes are expressed mainly during
plant reproductive development, where they play important roles in controlling
floral organ initiation and identity. Few previous reports have investigated the
functions of MADS-box transcription factors expressed in vegetative tissues. Here
we describe the functional characterization of a rice AG-like MADS-box protein,
OsMDP1 (Oryza sativa MADS-domain-containing protein 1). A partial cDNA encoding a
MADS-box domain was identified via high-throughput screening of rice
brassinolide-regulated genes, and the full-length cDNA was subsequently isolated
via screening of a cDNA library constructed from rice materials at tillering
stage. Expression pattern analyses indicated that OsMDP1 is transcribed mainly in
vegetative tissues, including the mature leaf, coleoptile, root-elongation zone,
culm internode, and especially the joint region between the leaf blade and
sheath. Further studies revealed that transcription of OsMDP1 is stimulated by
darkness and suppressed by brassinolide treatment. OsMDP1 deficiency resulted in
shortened primary roots, elongated coleoptiles and enhanced lamina joint
inclinations. Moreover, transgenic plants showed hypersensitivities to exogenous
brassinolide in terms of lamina joint inclination and coleoptile elongation.
OsMDP1 deficiency resulted in enhanced expression of OsXTR1, which encodes
xyloglucan endotransglycosylase, the cell-wall loosening enzyme necessary for
cell elongation, and modulated expressions of multiple genes involved in cell
signalling and gene transcription, indicating the key negative regulatory role of
OsMDP1 in BR signalling.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16827922 [PubMed - indexed for MEDLINE]

1242: Environ Biosafety Res. 2005 Oct-Dec;4(4):243-51. Epub 2006 Jun 22.

An ecological risk assessment of Cry1F maize pollen impact to pale grass blue
butterfly.

Wolt JD, Conlan CA, Majima K.

Iowa State University, Ames, IA, USA. jdwolt@iastate.edu

The intrinsic toxicity of lepidopteran-active Bt proteins necessitates assessment
of non-target risks associated with environmental release of transgenic crops
expressing these proteins. Principles of ecological risk assessment provide a
means for assessing non-target risks when information regarding exposure to the
toxin and species-specific effects are lacking. This is shown for the case of Bt
Cry1F maize release in Japan, where off-field pollen dissemination and effect on
butterfly species is of concern. The specific ecological entity of concern for
the assessment of the non-target impact of Cry1F maize pollen was Yamato-shijimi
(pale grass blue butterfly), Pseudozizeeria maha (Kollar), a commonly occurring,
susceptible species. Yamato-shijimi is widely adapted in Japan where it occurs in
both rural and metropolitan settings, corresponding to the distribution and
habitat of katabami (Oxalis corniculata (L.)), the larval host plant. The
northern extent of Yamato-shijimi habitat lies to the south of major maize
production regions in Japan, but exposure may occur elsewhere where maize and
Yamato-shijimi co-occur. Screening level assessment of potential adverse effects
to Yamato-shijimi in the field environment considered the probability for
spatial-temporal co-occurrence of the life stages of concern (1st and 2nd
instars) and the stressor (Cry1F protein expressed in maize pollen) at
environmentally relevant concentrations. In the event of exposure to maize
pollen, early instars of Yamato-shijimi feed exclusively on the underside of
katabami leaves, which further limits the portion of the butterfly population
that would be exposed. Projected levels of exposure to Cry1F pollen are below the
toxicity level of concern and, thus, indicate negligible risk. Most sensitive
species characterization (intergenera sensitivity) similarly shows negligible
risk to other Japanese butterfly species of concern when distributed beyond the
maize field or field margin.

PMID: 16827552 [PubMed - indexed for MEDLINE]

1243: Environ Biosafety Res. 2005 Oct-Dec;4(4):217-22. Epub 2006 Jun 22.

Monitoring the escape of transgenic oilseed rape around Japanese ports and
roadsides.

Saji H, Nakajima N, Aono M, Tamaoki M, Kubo A, Wakiyama S, Hatase Y, Nagatsu M.

Environmental Biology Division, National Institute for Environmental Studies,
16-2 Onogawa, Tsukuba, 305-8506, Japan. hsaji@nies.go.jp

An investigation was carried out to monitor the escape and spread of oilseed rape
(Brassica napus) transgenic plants and the introgression of transgenes to its
closely related feral species in Japan. We screened a total of about 7500 feral
B. napus, 300 B. rapa, and 5800 B. juncea seedlings from maternal plants in 143
locations at several ports, roadsides, and riverbanks. The presence of
glufosinate-resistance or glyphosate-resistance transgenes in these seedlings was
confirmed by means of herbicide treatments and also immunochemical and DNA
analyses. B. napus plants with herbicide-resistant transgenic seeds were found at
five of six major ports and along two of four sampled roadsides in the Kanto
District. Transgenic oilseed rape plants have not been commercially cultivated in
Japan, suggesting that the transgenes would probably have come from imported
transgenic seeds that were spilled during transportation to oilseed processing
facilities. No transgenes were detected in seeds collected from B. napus plants
growing along riverbanks in the Kanto District or in seeds from closely related
species (B. rapa and B. juncea). To our knowledge, this is the first published
example of feral, transgenic populations occurring in a nation where the
transgenic crop has not been cultivated commercially.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16827549 [PubMed - indexed for MEDLINE]

1244: Environ Biosafety Res. 2005 Oct-Dec;4(4):197-208; discussion 209-15. Epub
2006 Jun 22.

Detecting (trans)gene flow to landraces in centers of crop origin: lessons from
the case of maize in Mexico.

Cleveland DA, Soleri D, Cuevas FA, Crossa J, Gepts P.

Environmental Studies Program, University of California, Santa Barbara, CA
93106-4160, USA. cleveland@es.ucsb.edu

There is much discussion of the probability of transgene flow from transgenic
crop varieties to landraces and wild relatives in centers of origin or diversity,
and its genetic, ecological, and social consequences. Without costly research on
the variables determining gene flow, research on transgene frequencies in
landrace (or wild relative) populations can be valuable for understanding
transgene flow and its effects. Minimal research requirements include (1)
understanding how farmer practices and seed systems affect landrace populations,
(2) sampling to optimize Ne/n (effective/census population size), (3) minimizing
variance at all levels sampled, and (4) using Ne to calculate binomial
probabilities for transgene frequencies. A key case is maize in Mexico. Two
peer-reviewed papers, based on landrace samples from the Sierra Juö¡rez region of
Oaxaca, Mexico, reached seemingly conflicting conclusions: transgenes are present
(Quist and Chapela, 2001, Nature 414: 541-543; 2002, Nature 416: 602) or
"detectable transgenes" are absent (Ortiz-Garcö­a et al., 2005, Proc. Natl. Acad.
Sci. USA 102: 12338-12343 and 18242). We analyzed these papers using information
on Oaxacan maize seed systems and estimates of Ne. We conclude that if Quist and
Chapela's results showing presence are accepted, Ortiz-Garcö­a et al.'s
conclusions of no evidence of transgenes at detectable levels or for their
introgression into maize landraces in the Sierra de Juö¡rez of Oaxaca are not
scientifically justified. This is because their samples are not representative,
and their statistical analysis is inconclusive due to using n instead of Ne.
Using estimates of Ne based on Ortiz-Garcö­a et al.'s n, we estimate that
transgenes could be present in maize landraces in the Sierra Juö¡rez region at
frequencies of approximately 1-4%, and are more likely to be present in the 90%
of Oaxacan landrace area that is not mountainous. Thus, we have no scientific
evidence of maize transgene presence or absence in recent years in Mexico, Oaxaca
State, or the Sierra Juö¡rez region.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16827547 [PubMed - indexed for MEDLINE]

1245: Transgenic Res. 2006 Oct;15(5):627-36. Epub 2006 Jul 7.

High level expression of tissue-nonspecific alkaline phosphatase in the milk of
transgenic rabbits.

Bodrogi L, Brands R, Raaben W, Seinen W, Baranyi M, Fiechter D, Bosze Z.

Department of Animal Biology, Agricultural Biotechnology Center, P.O.B. 411,
H-2100 Gö¶dö¶llo, Hungary.

Alkaline phosphatase is a promising therapeutic agent in the Gram-negative
bacterial lipopolysaccharide (LPS) mediated acute and chronic diseases. Contrary
to other alkaline phosphatase isozymes, purified tissue-nonspecific alkaline
phosphatase (TNAP) is not available in large quantities from tissue sources,
which would enable to analyse its efficacy in animal sepsis models. Two
transgenic rabbit lines were created by pronuclear microinjection with the whey
acidic protein promoter-humanTNAP minigene (WAP-hTNAP). Lactating females of both
lines produced biologically active human TNAP. As indicated by fractionation of
milk samples the recombinant alkaline phosphatase was associated with the
membrane of milk fat globules. Alkaline phosphatase enzymatic activity was two
orders of magnitude higher compared to normal human serum levels. The
demonstration that this TNAP is physiologically active would provide the clue to
use transgenic animals as bioreactor for bulk production of the human
tissue-nonspecific alkaline phosphatase in milk. This may be a valuable and
possibly viable option with important implication in attenuating LPS mediated
inflammatory responses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16826424 [PubMed - indexed for MEDLINE]

1246: Nat Genet. 2006 Aug;38(8):876-8. Epub 2006 Jul 2.

Epigenetic asymmetry of imprinted genes in plant gametes.

Gutiö©rrez-Marcos JF, Costa LM, Dal Prö  M, Scholten S, Kranz E, Perez P, Dickinson
HG.

Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1
3RB, UK. jose.gutierrez@plants.ox.ac.uk

Plant imprinted genes show parent-of-origin expression in seed endosperm, but
little is known about the nature of parental imprints in gametes before
fertilization. We show here that single differentially methylated regions (DMRs)
correlate with allele-specific expression of two maternally expressed genes in
the seed and that one DMR is differentially methylated between gametes. Thus,
plants seem to have developed similar strategies as mammals to epigenetically
mark imprinted genes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16823380 [PubMed - indexed for MEDLINE]

1247: Proc Biol Sci. 2006 Aug 7;273(1596):1921-8.

Weed seed resources for birds in fields with contrasting conventional and
genetically modified herbicide-tolerant crops.

Gibbons DW, Bohan DA, Rothery P, Stuart RC, Haughton AJ, Scott RJ, Wilson JD,
Perry JN, Clark SJ, Dawson RJ, Firbank LG.

RSPB, UK Headquarters, The Lodge, Sandy, Bedfordshire SG19 2DL, UK.
david.gibbons@rspb.org.uk

The UK Farm Scale Evaluations (FSEs) have shown that the use of broad spectrum
herbicides on genetically modified herbicide-tolerant (GMHT) crops can have
dramatic effects on weed seed production compared to management of conventional
varieties. Here, we use FSE data and information on bird diets to determine how
GMHT cropping might change the food resources available to farmland birds. More
than 60 fields of each of four crops, spring- and winter-sown oilseed rape, beet
and maize, were split, one half being sown with a conventional variety, the other
with a GMHT variety. Seed rain from weeds known to be important in the diets of
17 granivorous farmland bird species was measured under the two treatments. In
beet and spring oilseed rape, rain of weed seeds important in the diets of 16
bird species was significantly reduced in GMHT compared to conventional halves;
for no species did it increase. In winter oilseed rape, rain of weed seeds
important in the diets of 10 species was significantly reduced in GMHT halves;
for only one species did it increase significantly. By contrast, in maize, rain
of weed seeds important in the diets of seven species was significantly greater
in GMHT halves; for no species was it reduced. Treatment effects for the total
weed seed energy available to each bird species were very similar to those for
seed rain alone. Measuring the effects on individual bird species was outside the
scope of this study. Despite this, these results suggest that should beet, spring
and winter rape crops in the UK be largely replaced by GMHT varieties and managed
as in the FSEs, this would markedly reduce important food resources for farmland
birds, many of which declined during the last quarter of the twentieth century.
By contrast, GMHT maize would be beneficial to farmland birds.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16822753 [PubMed - indexed for MEDLINE]

1248: Planta. 2007 Jan;225(2):277-86. Epub 2006 Jul 5.

Development of Cucumber mosaic virus as a vector modifiable for different host
species to produce therapeutic proteins.

Matsuo K, Hong JS, Tabayashi N, Ito A, Masuta C, Matsumura T.

National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1
Tsukisamu-Higashi, Sapporo, Japan.

We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially
for production of pharmaceutical proteins. The CMV vector is a vector modifiable
for different host plants and does not require further engineering steps. CMV
contains three genomic RNA molecules (RNAs 1-3) necessary for infectivity. With
this system, instead of creating different vector constructs for each plant we
use, we take advantage of the formation of pseudrecombinants between two CMV
isolates by simply reassembling a vector construct (RNA 2 base) and an RNA
molecule containing the host determinant (mostly RNA 3). In this study, the gene
for acidic fibroblast growth factor (aFGF), one of the human cytokines, was
cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based
vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up
to 5-8% of the total soluble protein. The tobacco-produced aFGF was purified, and
its biological activity was confirmed. Using this system, which provides a
versatile and viable strategy for the production of therapeutic proteins in
plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and
Arabidopsis thaliana.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16821041 [PubMed - indexed for MEDLINE]

1249: Funct Integr Genomics. 2006 Oct;6(4):263-84. Epub 2006 Jul 25.

Salt stress response in rice: genetics, molecular biology, and comparative
genomics.

Sahi C, Singh A, Kumar K, Blumwald E, Grover A.

Department of Plant Molecular Biology, University of Delhi South Campus, Benito
Juarez Road, Dhaula Kuan, New Delhi 110021, India.

Significant progress has been made in unraveling the molecular biology of rice in
the past two decades. Today, rice stands as a forerunner amongst the cereals in
terms of details known on its genetics. Evidence show that salt tolerance in
plants is a quantitative trait. Several traditional cultivars, landraces, and
wild types of rice like Pokkali, CSR types, and Porteresia coarctata appear as
promising materials for donation of requisite salt tolerance genes. A large
number of quantitative trait loci (QTL) have been identified for salt tolerance
in rice through generation of recombinant inbred lines and are being mapped using
different types of DNA markers. Salt-tolerant transgenic rice plants have been
produced using a host of different genes and transcript profiling by micro- and
macroarray-based methods has opened the gates for the discovery of novel salt
stress mechanisms in rice, and comparative genomics is turning out to be a
critical input in this respect. In this paper, we present a comprehensive review
of the genetic, molecular biology, and comparative genomics effort towards the
generation of salt-tolerant rice. From the data on comprehensive transcript
expression profiling of clones representing salt-stress-associated genes of rice,
it is shown that transcriptional and translational machineries are important
determinants in controlling salt stress response, and gene expression response in
tolerant and susceptible rice plants differs mainly in quantitative terms.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't
Review

PMID: 16819623 [PubMed - indexed for MEDLINE]

1250: Mol Cells. 2006 Jun 30;21(3):401-10.

Plastid transformation in the monocotyledonous cereal crop, rice (Oryza sativa)
and transmission of transgenes to their progeny.

Lee SM, Kang K, Chung H, Yoo SH, Xu XM, Lee SB, Cheong JJ, Daniell H, Kim M.

School of Agricultural Biotechnology, Seoul National University, Seoul 151-921,
Korea.

The plastid transformation approach offers a number of unique advantages,
including high-level transgene expression, multi-gene engineering, transgene
containment, and a lack of gene silencing and position effects. The extension of
plastid transformation technology to monocotyledonous cereal crops, including
rice, bears great promise for the improvement of agronomic traits, and the
efficient production of pharmaceutical or nutritional enhancement. Here, we
report a promising step towards stable plastid transformation in rice. We
produced fertile transplastomic rice plants and demonstrated transmission of the
plastid-expressed green fluorescent protein (GFP) and aminoglycoside
3'-adenylyltransferase genes to the progeny of these plants. Transgenic
chloroplasts were determined to have stably expressed the GFP, which was
confirmed by both confocal microscopy and Western blot analyses. Although the
produced rice plastid transformants were found to be heteroplastomic, and the
transformation efficiency requires further improvement, this study has
established a variety of parameters for the use of plastid transformation
technology in cereal crops.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16819304 [PubMed - indexed for MEDLINE]

1251: Clin Mol Allergy. 2006 Jul 4;4:10.

Evaluation of the sensitization rates and identification of IgE-binding
components in wild and genetically modified potatoes in patients with allergic
disorders.

Lee SK, Ye YM, Yoon SH, Lee BO, Kim SH, Park HS.

Department of Internal Medicine, College of Medicine, Dong-A University, Busan,
Korea. skleeai@daunet.donga.ac.kr

BACKGROUND: The potato is one of the most common types of genetically modified
(GM) food. However, there are no published data evaluating the impact of genetic
manipulations on the allergenicity of GM potatoes. To compare the allergenicity
of GM potatoes with that of wild-type potatoes using in vivo and in vitro methods
in adult allergy patients sensitized to potatoes. METHODS: A total of 1886
patients with various allergic diseases and 38 healthy controls participated in
the study. Skin-prick testing and IgE-ELISA were carried out with extracts
prepared from wild-type and GM potatoes. An ELISA inhibition test was used to
confirm the binding specificity. IgE-binding components in extracts from the two
types of potato were identified by SDS-PAGE and IgE-immunoblotting. The effects
of digestive enzymes and heat on the allergenicity of the extracts was evaluated
by preincubating the potatoes with or without simulated gastric and intestinal
fluids in the absence or presence of heat. RESULTS: Positive responses (ratio of
the wheal size induced by the allergen to that induced by histamine (A/H) > or =
2+) to wild-type or GM potato extracts, as demonstrated by the skin-prick test,
were observed in 108 patients (5.7%). Serum-specific IgE was detected in 0-88% of
subjects who tested positively. ELISA inhibition tests indicated significant
inhibition when extract from each type of potato was added. IgE-immunoblot
analysis demonstrated the presence of 14 IgE-binding components within the
wild-type potato and 9 within the GM potato. Furthermore, a common 45-kDa binding
component that yielded similar IgE-binding patterns was noted in more than 80% of
the reactions using sera from patients sensitized to wild-type or GM potato.
Exposure to simulated gastric fluid and heat treatment similarly inhibited IgE
binding by extracts from wild-type and GM potatoes, whereas minimal changes were
obtained following exposure of the extracts to simulated intestinal fluid.
CONCLUSION: Our results strongly suggest that genetic manipulation of potatoes
does not increase their allergenic risk. The sensitization rate of adult allergy
patients to both types of extract was 5.7%, and a common major allergen (45 kDa)
was identified.

PMID: 16817976 [PubMed]

1252: Plant Cell Physiol. 2006 Aug;47(8):1102-11. Epub 2006 Jul 2.

RNAi knock-down of ENOD40s leads to significant suppression of nodule formation
in Lotus japonicus.

Kumagai H, Kinoshita E, Ridge RW, Kouchi H.

National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki,
305-8602 Japan.

ENOD40 is one of the most intriguing early nodulin genes that is known to be
induced very early in response to interaction of legume plants with symbiotic
Rhizobium bacteria, but its function in the nodulation process is still not
known. Lotus japonicus has two ENOD40 genes: LjENOD40-1 is abundantly induced in
very early stages of bacterial infection or Nod factor application, whereas
LjENOD40-2 is abundantly expressed only in mature nodules. We generated
transgenic lines of L. japonicus with an RNAi (RNA interference) construct that
expresses hairpin double-stranded RNA for LjENOD40-1 to induce sequence-specific
RNA silencing. In the transgenic plants, expression of both LjENOD40-1 and -2 was
significantly reduced, and no accumulation of ENOD40 transcripts was detected
upon Mesorhizobium loti inoculation. The transgenic plants exhibited very poor
nodulation (only 0-2 nodules per plant) and could not grow well without
additional nitrogen supply. Analysis of segregation in the T(2) progeny indicated
that the suppression of nodulation is perfectly linked with the presence of the
transgene. Microscopic observation of the infection process using lacZ-labeled M.
loti, together with expression analysis of infection-related nodulin genes,
demonstrated that ENOD40 knock-down did not inhibit the initiation of the
bacterial infection process. In contrast, nodule primordium initiation and
subsequent nodule development were significantly suppressed in the transgenic
plants. These results clearly indicate that ENOD40 is required for nodule
initiation and subsequent organogenesis, but is not involved in early infection
events.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16816411 [PubMed - indexed for MEDLINE]

1253: Plant Physiol. 2006 Sep;142(1):233-44. Epub 2006 Jun 30.

Expression and functional analyses of the plastid lipid-associated protein CHRC
suggest its role in chromoplastogenesis and stress.

Leitner-Dagan Y, Ovadis M, Shklarman E, Elad Y, Rav David D, Vainstein A.

Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Hebrew
University of Jerusalem, Rehovot 76100, Israel.

Chromoplastogenesis during flower development and fruit ripening involves the
dramatic overaccumulation of carotenoids sequestered into structures containing
lipids and proteins called plastid lipid-associated proteins (PAPs). CHRC, a
cucumber (Cucumis sativus) PAP, has been suggested to be transcriptionally
activated in carotenoid-accumulating flowers by gibberellin (GA). Mybys, a
MYB-like trans-activator identified here, may represent a
chromoplastogenesis-related factor: Its expression is flower specific and
parallels that of ChrC during flower development; moreover, as revealed by stable
ectopic and transient-expression assays, it specifically trans-activates ChrC
promoter in flowers accumulating carotenoids and flavonoids. A detailed
dissection of ChrC promoter revealed a GA-responsive element, gacCTCcaa, the
mutation of which abolished ChrC activation by GA. This cis-element is different
from the GARE motif and is involved in ChrC activation probably via negative
regulation, similar to other GA-responsive systems. The GA responsiveness and
MYBYS floral activation of the ChrC promoter do not overlap with respect to
cis-elements. To study the functionality of CHRC, which is activated in
vegetative tissues similar to other PAPs by various biotic and abiotic stresses,
we employed a tomato (Lycopersicon esculentum) plant system and generated
RNAi-transgenic lines with suppressed LeCHRC. Transgenic flowers accumulated
approximately 30% less carotenoids per unit protein than controls, indicating an
interrelationship between PAPs and flower-specific carotenoid accumulation in
chromoplasts. Moreover, the transgenic LeCHRC-suppressed plants were
significantly more susceptible to Botrytis cinerea infection, suggesting CHRC's
involvement in plant protection under stress conditions and supporting the
general, evolutionarily preserved role of PAPs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16815957 [PubMed - indexed for MEDLINE]

1254: Plant Physiol. 2006 Aug;141(4):1414-24. Epub 2006 Jun 30.

Overexpression of a protein phosphatase 2C from beech seeds in Arabidopsis shows
phenotypes related to abscisic acid responses and gibberellin biosynthesis.

Reyes D, Rodrö­guez D, Gonzö¡lez-Garcö­a MP, Lorenzo O, Nicolö¡s G, Garcö­a-Martö­nez
JL, Nicolö¡s C.

Departamento de Fisiologö­a Vegetal, Centro Hispano-Luso de Investigaciones
Agrarias, Facultad de Biologö­a, Universidad de Salamanca, 37007 Salamanca, Spain.

A functional abscisic acid (ABA)-induced protein phosphatase type 2C (PP2C) was
previously isolated from beech (Fagus sylvatica) seeds (FsPP2C2). Because
transgenic work is not possible in beech, in this study we overexpressed this
gene in Arabidopsis (Arabidopsis thaliana) to provide genetic evidence on FsPP2C2
function in seed dormancy and other plant responses. In contrast with other PP2Cs
described so far, constitutive expression of FsPP2C2 in Arabidopsis, under the
cauliflower mosaic virus 35S promoter, produced enhanced sensitivity to ABA and
abiotic stress in seeds and vegetative tissues, dwarf phenotype, and delayed
flowering, and all these effects were reversed by gibberellic acid application.
The levels of active gibberellins (GAs) were reduced in 35S:FsPP2C2 plants,
although transcript levels of AtGA20ox1 and AtGA3ox1 increased, probably as a
result of negative feedback regulation, whereas the expression of GASA1 was
induced by GAs. Additionally, FsPP2C2-overexpressing plants showed a strong
induction of the Responsive to ABA 18 (RAB18) gene. Interestingly, FsPP2C2
contains two nuclear targeting sequences, and transient expression assays
revealed that ABA directed this protein to the nucleus. Whereas other plant PP2Cs
have been shown to act as negative regulators, our results support the hypothesis
that FsPP2C2 is a positive regulator of ABA. Moreover, our results indicate the
existence of potential cross-talk between ABA signaling and GA biosynthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16815952 [PubMed - indexed for MEDLINE]

1255: J Econ Entomol. 2006 Jun;99(3):927-30.

Effect of Bacillus thuringiensis cry3Bb1 protein on the feeding behavior and
longevity of adult western corn rootworms (Coleoptera: Chrysomelidae).

Nowatzki TM, Zhou X, Meinke LJ, Vaughn T, Siegfried BD.

Department of Entomology, University of Nebraska-Lincoln, Lincoln, NE 68583-0816,
USA.

The first transgenic corn hybrids expressing the Bacillus thuringiensis (Bt)
Cry3Bb1 protein to control corn rootworm (Diabrotica spp.) larvae were registered
for commercial use in 2003. This study was conducted to investigate the effect of
Cry3Bb1 protein in combination with a cucurbitacin bait on adult feeding and
longevity of both organophosphate-resistant and -susceptible western corn
rootworms, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae). In
choice and no-choice tests, possible repellency to the Bt protein was quantified
by comparing beetle consumption of cellulose disks treated with three
concentrations of Bt in combination with a feeding stimulant (Invite EC) to disks
treated with stimulant alone. A lethal-time assay also was conducted to examine
survival of beetles exposed to Bt protein in their diet. Results from these
assays indicate that adult rootworms are not significantly deterred by the
presence of Cry3Bb1 on the treated discs and that ingestion of toxin does not
adversely affect adult longevity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16813332 [PubMed - indexed for MEDLINE]

1256: J Econ Entomol. 2006 Jun;99(3):899-907.

Planting transgenic insecticidal corn based on economic thresholds: consequences
for integrated pest management and insect resistance management.

Crowder DW, Onstad DW, Gray ME.

Department of Natural Resources and Environmental Sciences, University of
Illinois, Urbana, IL 61801, USA.

A simulation model of the western corn rootworm, Diabrotica virgifera virgifera
LeConte, was used to investigate whether sampling and economic thresholds can
improve integrated pest management (IPM) and insect resistance management (IRM)
when transgenic insecticidal crops are used for insect pest management. When
transgenic corn killed at least 80% of susceptible larvae, the calculated
economic threshold increased linearly as the proportion of susceptible beetles
surviving the toxin increased. The use of economic thresholds slightly slowed the
evolution of resistance to transgenic insecticidal crops. In areas with or
without rotation-resistant western corn rootworm phenotypes, the use of sampling
and economic thresholds generated similar returns compared with strategies of
planting transgenic corn, Zea mays L., every season. Because transgenic crops are
extremely effective, farmers may be inclined to plant transgenic crops every
season rather than implementing costly and time-consuming sampling protocols.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16813328 [PubMed - indexed for MEDLINE]

1257: J Econ Entomol. 2006 Jun;99(3):893-8.

Resistance monitoring of Helicoverpa armigera (Lepidoptera: Noctuidae) to bt
insecticidal protein during 2001-2004 in China.

Wu K, Guo Y, Head G.

SKLBPI, Institute of Plant Protection, Chinese Academy of Agricultural Sciences,
Beijing 100094, People's Republic of China. wkm@caascose.net.cn

Susceptibility of Helicoverpa armigera (Hö¼bner) (Lepidoptera: Noctuidae) field
populations to the CrylAc toxin of Bacillus thuringiensis Berliner (Bt) were
monitored from 1997 to 2004 in China. During 2001-2004, 53 strains from the Bt
cotton planting region were sampled. The range of concentration producing 50%
inhibition of larval development to third instar (IC50) values among different
populations in 2001, 2002, 2003, and 2004 was 0.014-0.046, 0.010-0.062,
0.005-0.062, and 0.005-0.035 microg/ml, respectively. Diagnostic concentration
studies (IC99) showed that the percentage of individuals reaching third instar
ranged from 0 to 9.09%, with only four of the 53 tested populations showing
values above 0%. Considering these data, it was determined that the
susceptibility to CrylAc of the field populations sampled was not different from
the baseline in 1997, and no movement toward resistance among H. armigera
populations was apparent.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16813327 [PubMed - indexed for MEDLINE]

1258: J Econ Entomol. 2006 Jun;99(3):728-32.

Cannibalism of Helicoverpa zea (Lepidoptera: Noctuidae) from Bacillus
thuringiensis (Bt) transgenic corn versus non-Bt corn.

Chilcutt CF.

Department of Entomology, Texas A&M University Agricultural Research & Extension
Center, 10345 Agnes Street, Corpus Christi, TX 78406, USA.

Because of the importance of cannibalism in population regulation of Helicoverpa
zea (Boddie) (Lepidoptera: Noctuidae) in corn, Zea mays L., it is useful to
understand the interactions between Bacillus thuringiensis (Bt) transgenic corn
and cannibalism. To determine the effects of Bt corn on cannibalism in H. zea,
pairs of the same or different instars were taken from Bt or non-Bt corn and
placed on artificial diet in proximity. Cannibalism occurred in 91% of pairs and
was approximately 7% greater for pairs of larvae reared from Bt transgenic corn
(95%) than from non-Bt corn (88%). Also, first instar by first instar pairs had a
lower rate of cannibalism than other pairs. Time until cannibalism was not
different for larvae from Bt corn versus non-Bt corn. Pupation rate of cannibals
and surviving victims was not different for pairs from Bt corn versus non-Bt
corn. Finally, cannibalism increased pupation rate of cannibals from both Bt and
non-Bt corn by approximately 23 and 12%, respectively, although the increases
were not significant. Thus, negative effects of Bt on larvae were compensated by
increased cannibalism in comparison with larvae reared on non-Bt corn, which
increased larval survival to levels comparable with larvae reared on non-Bt
plants.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16813305 [PubMed - indexed for MEDLINE]

1259: J Econ Entomol. 2006 Jun;99(3):722-7.

Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) larval feeding
behavior on transgenic maize (MON 863) and its isoline.

Clark PL, Vaughn TT, Meinke LJ, Molina-Ochoa J, Foster JE.

Monsanto Company, 800 North Lindbergh Blvd., St. Louis, MO 63167, USA.

Diabrotica species (Coleoptera: Chrysomelidae) larval behavior studies have posed
a challenge to researchers because of the subterranean life cycle of this pest.
To fully understand how the western corn rootworm, Diabrotica virgifera virgifera
LeConte, injures the maize, Zea mays L., root system, its behavior must be
studied. For example, larvae that can detect an area of the root that has a lower
amount of toxin, whether from an insecticide or a transgenic maize plant, have an
increased chance of survival. This study assessed D. v. virgifera larval feeding
behavior on rootworm-susceptible maize and maize containing a
biotechnology-derived trait (MON 863) with resistance to D. v. virgifera first
instar feeding. Maize plants were grown in a medium that allowed for direct
observation and measurements during feeding of larval stadia. Neonates were
placed on maize seedlings, and data were taken at 3, 6, 9, and 12 d
postinfestation on resistant and susceptible maize. On rootworm-susceptible
maize, neonate larvae aggregated at the root tips and began actively feeding, and
then they moved to older root tissue. Conversely, some larvae that ingested Cry
3Bb1 from the resistant maize exhibited no movement. Other larvae on the
resistant maize moved continuously, sampling root hairs or root tissue but not
actively feeding. The continuously moving larvae had visibly empty guts,
suggesting possible nonpreference for the resistant root. This study contributes
to our understanding of D. v. virgifera larval behavior and provides insight into
questions surrounding the potential evolution of behavioral and biochemical
resistance to Cry3Bb1.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16813304 [PubMed - indexed for MEDLINE]

1260: Plant Cell Rep. 2006 Dec;25(12):1362-8. Epub 2006 Jun 30.

Sequence stability of the T-DNA - plant junctions in tissue culture in
Arabidopsis transgenic lines.

Papazova N, Windels P, Depicker A, Taverniers I, Roldan-Ruiz I, Milcamps A, Van
Bockstaele E, Van Den Eede G, De Loose M.

Unit Technology and Food, Institute for Agricultural and Fisheries Research,
9820, Merelbeke, Belgium. nina.papazova@ilvo.vlaanderen.be

The stability of the inserted transgenes and particularly the junction regions of
transgenic events is a concern of food labeling, traceability and post release
monitoring, as these regions are used for development of event-specific DNA-based
detection methods. During the standard agricultural breeding practices, the
transgenic lines can be exposed to completely different conditions than those in
the laboratory environment. Some of these conditions have the potential to affect
the stability of the transgenic locus and the surrounding DNA. As tissue culture
is recognized as a stressful and mutagenic factor, we have analyzed the effect of
this process on the stability of the junction regions at nucleotide level in five
Arabidopsis thaliana transgenic lines in comparison with the respective
integration loci in ColO and C24 ecotypes. No indication of any kind of
alteration at nucleotide level of the junctions was found. The relevance of the
stability of the plant-T-DNA junction regions for application of the DNA-based
methods in commercial transgenic plants is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16810524 [PubMed - indexed for MEDLINE]

1261: J Biosci. 2006 Jun;31(2):255-63.

Overexpression of GbERF confers alteration of ethylene-responsive gene expression
and enhanced resistance to Pseudomonas syringae in transgenic tobacco.

Qin J, Zuo K, Zhao J, Ling H, Cao Y, Qiu C, Li F, Sun X, Tang K.

Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology,
R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University,
Shaghai, People's Republic of China.

GbERF belongs to the ERF (ethylene responsive factor) family of transcription
factors and regulates the GCC-box containing pathogen-related (PR) genes in the
ethylene signal transduction pathway. To study the function of GbERF in the
process of biotic stress, transgenic tobacco plants expressing GbERF were
generated. Overexpression of GbERF did not change transgenic plant's phenotype
and endogenous ethylene level. However, the expression profile of some
ethylene-inducible GCC-box and non-GCC-box containing genes was altered, such as
PR1b, PR2, PR3, PR4, Osmotin, CHN50, ACC oxidase and ACC synthase genes. These
data indicate that the cotton GbERF could act as a transcriptional activator or
repressor to regulate the differential expression of ethylene-inducible genes via
GCC and non-GCC cis-elements. Moreover, the constitutive expression of GbERF in
transgenic tobacco enhanced the plant's resistance to Pseudomonas syringae pv
tabaci infection. In conclusion, GbERF mediates the expression of a wide array of
PR and ethylene-responsive genes and plays an important role in the plant's
response to biotic stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16809858 [PubMed - indexed for MEDLINE]

1262: J Biosci. 2006 Jun;31(2):235-46.

Genetic transformation of peanut (Arachis hypogaea L.) using cotyledonary node as
explant and a promoterless gus::nptII fusion gene based vector.

Anuradha TS, Jami SK, Datla RS, Kirti PB.

Department of Plant Sciences, University of Hyderabad, Hyderabad 500 046,India.

We have generated putative promoter tagged transgenic lines in Arachis hypogaea
cv JL-24 using cotyledonary node (CN) as an explant and a promoterless gus::nptII
bifunctional fusion gene mediated by Agrobacterium transformation. MS medium
fortified with 6-benzylaminopurine (BAP) at 4mg/l in combination with 0.1 mg/l
alpha -napthaleneacetic acid (NAA) was the most effective out of the various BAP
and NAA combinations tested in multiple shoot bud formation. Parameters enhancing
genetic transformation viz. seedling age, Agrobacterium genetic background and
co-cultivation periods were studied by using the binary vector p35SGUSINT.
Genetic transformation with CN explants from 6-day-old seedlings co-cultivated
with Agrobacterium GV2260 strain for 3 days resulted in high kanamycin resistant
shoot induction percentage (45%); approximately 31% transformation frequency was
achieved with p35S GUSINT in beta-glucuronidase (GUS) assays. Among the in vivo
GUS fusions studied with promoterless gus::nptII construct, GUS-positive sectors
occupied 38% of the total transient GUS percentage. We have generated over 141
putative T 0 plants by using the promoterless construct and transferred them to
the field. Among these, 82 plants survived well in the green house and 5 plants
corresponding to 3.54% showed stable integration of the fusion gene as evidenced
by GUS, polymerase chain reaction (PCR) and Southern blot analyses. Twenty-four
plants were positive for GUS showing either tissue-specific expression or blue
spots in at least one plant part. The progeny of 15 T 0 plants indicated
Mendelian inheritance pattern of segregation for single-copy integration. The
tissue-specific GUS expression patterns were more or less similar in both T 0 and
corresponding T 1 progeny plants. We present the differential patterns of GUS
expression identified in the putative promoter-tagged transgenic lines in the
present communication.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16809856 [PubMed - indexed for MEDLINE]

1263: Phytochemistry. 2006 Jul;67(14):1442-54. Epub 2006 Jun 30.

OsBLE3, a brassinolide-enhanced gene, is involved in the growth of rice.

Yang G, Nakamura H, Ichikawa H, Kitano H, Komatsu S.

National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba 305-8602,
Japan.

Brassinosteroids (BRs) are a group of plant hormones involved in a wide range of
plant growth and developmental processes. To investigate the mechanism of BR
action in monocots, a brassinolide (BL) upregulated gene designated OsBLE3 was
identified, cloned and characterized in rice. It was mainly expressed in roots
and leaf sheaths with levels of expression directly dependent on the dose of BL.
In situ hybridization detected OsBLE3 mRNA in the shoot apical meristem, organ
primordia and vascular tissue. Furthermore, its expression was enhanced by
co-treatment with BL and low concentrations of IAA. These results, and the
existence of auxin response elements in the 5'-flanking region of the OsBLE3
gene, indicate that OsBLE3 expression is under control of both BR and auxin. The
GUS reporter gene driven by a 2277 bp OsBLE3 putative promoter was mainly
expressed in vascular tissues, branch root primordia and was responsive to
exogenous BL treatment. OsBLE3 transcript levels were greatly reduced in brd1
plants, a BL deficient mutant, compared to the wild type control. In OsBRI1
antisense transgenic rice and OsBLE3, the BR-insensitive mutant expression of
OsBLE3 in response to exogenous BL treatment was significantly lower compared to
that in control plants transformed with a vacant vector. Reduced OsBLE3
expression and growth retardation was also observed in OsBLE3 antisense
transgenic rice plants. Internode cell length of the OsBLE3 antisense transgenic
lines was about 70% of that in the vacant vector transformed control lines. These
results suggest that OsBLE3 is involved in cell elongation in rice through dual
regulation by BL and IAA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16808934 [PubMed - indexed for MEDLINE]

1264: FEBS Lett. 2006 Jul 10;580(16):3980-8. Epub 2006 Jun 21.

Introgression of a novel salt-tolerant L-myo-inositol 1-phosphate synthase from
Porteresia coarctata (Roxb.) Tateoka (PcINO1) confers salt tolerance to
evolutionary diverse organisms.

Das-Chatterjee A, Goswami L, Maitra S, Dastidar KG, Ray S, Majumder AL.

Plant Molecular and Cellular Genetics, Bose Institute, P-1/12, C I T Scheme VIIM,
Kolkata 700 054, India.

We have previously demonstrated that introgression of PcINO1 gene from Porteresia
coarctata (Roxb.) Tateoka, coding for a novel salt-tolerant L-myo-inositol
1-phosphate synthase (MIPS) protein, confers salt tolerance to transgenic tobacco
plants (Majee, M., Maitra, S., Dastidar, K.G., Pattnaik, S., Chatterjee, A.,
Hait, N.C., Das, K.P. and Majumder, A.L. (2004) A novel salt-tolerant
L-myo-inositol-1-phosphate synthase from Porteresia coarctata (Roxb.) Tateoka, a
halophytic wild rice: molecular cloning, bacterial overexpression,
characterization, and functional introgression into tobacco-conferring
salt-tolerance phenotype. J. Biol. Chem. 279, 28539-28552). In this communication
we have shown that functional introgression of the PcINO1 gene confers
salt-tolerance to evolutionary diverse organisms from prokaryotes to eukaryotes
including crop plants albeit to a variable extent. A direct correlation between
unabated increased synthesis of inositol under salinity stress by the PcINO1 gene
product and salt tolerance has been demonstrated for all the systems pointing
towards the universality of the application across evolutionary divergent taxa.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16806195 [PubMed - indexed for MEDLINE]

1265: Plant J. 2006 Jun;46(6):1059-72.

Establishment of a patterned GAL4-VP16 transactivation system for discovering
gene function in rice.

Liang D, Wu C, Li C, Xu C, Zhang J, Kilian A, Li X, Zhang Q, Xiong L.

National Key Laboratory of Crop Genetic Improvement, National Center of Plant
Gene Research (Wuhan), Huazhong Agricultural University, Wuhan 430070, China.

A binary GAL4-VP16-UAS transactivation system has been established in rice (Oryza
sativa L.) in this study for the discovery of gene functions. This binary system
consists of two types of transgenic lines, pattern lines and target lines. The
pattern lines were produced by transformation of Zhonghua 11, a japonica
cultivar, with a construct consisting of the transactivator gene GAL4-VP16
controlled by a minimal promoter and the GUSplus reporter controlled by the
upstream activation sequence (UAS; cis-element to GAL4). Target lines were
generated by transformation of Zhonghua 11 with constructs carrying the EGFP
reporter and target genes of interest, both controlled by the UAS but in opposite
directions. Hybrid plants were obtained by crossing target lines of 10 putative
transcription factor genes from rice with six pattern lines showing expression in
anther, stigma, palea, lemma and leaves. The EGFP and target genes perfectly
co-expressed in hybrid plants with the same expression patterns as in the pattern
lines. Various phenotypic changes, such as delayed flowering, multiple pistils,
dwarfism, narrow and droopy leaves, reduced tillers, growth retardation and
sterility, were induced as a result of the expression of the target genes. It is
concluded that this transactivation system can provide a useful tool in rice to
unveil latent functions of unknown or known genes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16805737 [PubMed - indexed for MEDLINE]

1266: Plant J. 2006 Jun;46(6):1032-44.

Characterization and functional analysis of ABSCISIC ACID INSENSITIVE3-like genes
from Physcomitrella patens.

Marella HH, Sakata Y, Quatrano RS.

Department of Biology, Washington University, 1 Brookings Drive, St Louis, MO
63130, USA.

Although the moss Physcomitrella patens is known to respond to abscisic acid
(ABA) by activating gene expression, the transcriptional components involved have
not been characterized. Initially, we used the ABA-responsive Em promoter from
wheat linked to beta-glucuronidase (GUS) to determine whether ABI3/VP1,
transcriptional regulators in the ABA-signaling pathway in angiosperms, were
similarly active in the ABA response of P. patens. We show by particle
bombardment that ABI3 and VP1 affect Em-GUS expression in P. patens in a manner
similar to angiosperms. We also show the involvement of ABI1 in the pathway,
utilizing the abi1-1 mutant allele. We isolated three ABI3-like genes from P.
patens. Using an Em-like ABA-responsive promoter from P. patens (PpLea1), we
demonstrate that PpABI3A, only in the presence of ABA, strongly enhances
PpLea1-GUS expression in P. patens. PpABI3A also enhances ABA-induced Em-GUS
expression in P. patens. In barley aleurone, PpABI3A transactivates Em-GUS but to
a lesser extent than VP1 and ABI3. PpABI3A:GFP is localized to the nucleus of
both protonemal cells and barley aleurone, indicating that the nuclear
localization signals are conserved. We show that at least a part of the inability
of PpABI3A to fully complement the phenotypes of the Arabidopsis abi3-6 mutant is
due to a weak interaction between PpABI3A and the bZIP transcription factor ABI5,
as assayed functionally in barley aleurone and physically in the yeast-two-hybrid
assay. Our data clearly demonstrate that P. patens will be useful for comparative
structural and functional studies of components in the ABA-response pathway such
as ABI3.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16805735 [PubMed - indexed for MEDLINE]

1267: Plant J. 2006 Jun;46(6):971-83.

Functional and signaling mechanism analysis of rice CRYPTOCHROME 1.

Zhang YC, Gong SF, Li QH, Sang Y, Yang HQ.

National Key Laboratory of Plant Molecular Genetics, Institute of Plant
Physiology and Ecology, Shanghai, China.

Cryptochromes (CRY) are blue-light photoreceptors that mediate various light
responses, such as inhibition of hypocotyl elongation, enhancement of cotyledon
expansion, anthocyanin accumulation and stomatal opening in Arabidopsis. The
signaling mechanism of Arabidopsis CRY is mediated through direct interaction
with COP1, a negative regulator of photomorphogenesis. CRY has now been
characterized in tomato, pea, moss and fern, but its function in monocots is
largely unknown. Here we report the function and basic signaling mechanism of
rice cryptochrome 1 (OsCRY1). Overexpresion of OsCRY1b resulted in a blue
light-dependent short hypcotyl phenotype in Arabidopsis, and a short coleoptile,
leaf sheath and leaf blade phenotype in rice (Oryza sativa). On fusion with
beta-glucuronidase (GUS), the C-terminal domain of either OsCRY1a (OsCCT1a) or
OsCRY1b (OsCCT1b) mediated a constitutive photomorphogenic (COP) phenotype in
both Arabidopsis and rice, whereas OsCCT1b mutants corresponding to missense
mutations in previously described Arabidopsis cry1 alleles failed to confer a COP
phenotype. Yeast two-hybrid and subcellular co-localization studies demonstrated
that OsCRY1b interacted physically with rice COP1 (OsCOP1). From these results,
we conclude that OsCRY1 is implicated in blue-light inhibition of coleoptile and
leaf elongation during early seedling development in rice, and that the signaling
mechanism of OsCRY1 involves direct interaction with OsCOP1.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16805731 [PubMed - indexed for MEDLINE]

1268: J Exp Bot. 2006;57(10):2401-11. Epub 2006 Jun 27.

The strawberry gene FaGAST affects plant growth through inhibition of cell
elongation.

de la Fuente JI, Amaya I, Castillejo C, Sö¡nchez-Sevilla JF, Quesada MA, Botella
MA, Valpuesta V.

Departamento de Biologö­a Molecular y Bioquö­mica, Universidad de Mö¡laga, 29071
Mö¡laga, Spain.

The strawberry (Fragaria x ananassa) FaGAST gene encodes a small protein with 12
cysteine residues conserved in the C-terminal region similar to a group of
proteins identified in other species with diverse assigned functions such as cell
division, elongation, or elongation arrest. This gene is expressed in the fruit
receptacle, with two peaks during ripening at the white and the red-ripe stages,
both coincident with an arrest in the growth pattern. Expression is also high in
the roots but confined to the cells at the end of the elongation zone. Exogenous
application of gibberellin increased the transcript level of the FaGAST gene in
strawberry fruits. Ectopic expression of FaGAST in transgenic Fragaria vesca
under the control of the CaMV-35S promoter caused both delayed growth of the
plant and fruits with reduced size. The same growth defect was observed in
Arabidopsis thaliana plants overexpressing FaGAST. In addition, the transgenic
plants exhibited late flowering and low sensitivity to exogenous gibberellin.
Taken together, the expression pattern, the regulation by gibberellin, and the
transgenic phenotypes point to a role for FaGAST in arresting cell elongation
during strawberry fruit ripening.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16804055 [PubMed - indexed for MEDLINE]

1269: Plant Cell Rep. 2006 Nov;25(11):1166-73. Epub 2006 Jun 27.

Transgenic peas (Pisum sativum) expressing polygalacturonase inhibiting protein
from raspberry (Rubus idaeus) and stilbene synthase from grape (Vitis vinifera).

Richter A, Jacobsen HJ, de Kathen A, de Lorenzo G, Briviba K, Hain R, Ramsay G,
Kiesecker H.

Department of Molecular Genetics, University of Hannover, Herrenhö¤userstr 2,
30419, Hannover, Germany.

The pea (Pisum sativum L.) varieties Baroness (United Kingdome) and Baccara
(France) were transformed via Agrobacterium tumefaciens-mediated gene transfer
with pGPTV binary vectors containing the bar gene in combination with two
different antifungal genes coding for polygalacturonase-inhibiting protein (PGIP)
from raspberry (Rubus idaeus L.) driven by a double 35S promoter, or the stilbene
synthase (Vst1) from grape (Vitis vinifera L.) driven by its own
elicitor-inducible promoter. Transgenic lines were established and transgenes
combined via conventional crossing. Resveratrol, produced by Vst1 transgenic
plants, was detected using HPLC and the PGIP expression was determined in
functional inhibition assays against fungal polygalacturonases. Stable
inheritance of the antifungal genes in the transgenic plants was demonstrated.

PMID: 16802117 [PubMed - indexed for MEDLINE]

1270: Biotechnol Lett. 2006 Aug;28(16):1247-53. Epub 2006 Jun 27.

Chemical-induced autoexcision of selectable markers in elite tomato plants
transformed with a gene conferring resistance to lepidopteran insects.

Zhang Y, Li H, Ouyang B, Lu Y, Ye Z.

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural
University, Wuhan 430070, China.

Marker-free transgenic tomato plants harboring a synthetic Bacillus thuringiensis
endotoxin gene, cryIAc, were obtained by using a chemically regulated,
Cre/loxP-mediated site-specific DNA recombination system, in which the selectable
marker neomycin phosphotransferase gene flanked by two directly oriented loxP
sites was located between the cauliflower mosaic virus 35S promoter and a
promoterless cryIAc. Upon induction by 2 microM beta-estradiol, sequences
encoding the selectable marker and cre sandwiched by two loxP sites were excised
from the tomato genome, leading to activation of the downstream endotoxin gene
cryIAc with high expression levels as shown by Northern blot and ELISA assay
(250-790 ng g(-1) fresh wt) in T(1) generation. For transgenic line with single
transgenic loci, 15% of T(1) progenies were revealed marker-free. This
autoexcision strategy provides an effective approach to eliminate a selectable
marker gene from transgenic tomato, thus expediting the public acceptance of
genetically modified crop.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16802102 [PubMed - indexed for MEDLINE]

1271: BMC Plant Biol. 2006 Jun 26;6:13.

Metabolic engineering of potato tuber carotenoids through tuber-specific
silencing of lycopene epsilon cyclase.

Diretto G, Tavazza R, Welsch R, Pizzichini D, Mourgues F, Papacchioli V, Beyer P,
Giuliano G.

ENEA, Casaccia Research Center, PO Box 2400, Roma 00100AD, Italy.
gianfranco.diretto@casaccia.enea.it

BACKGROUND: Potato is a major staple food, and modification of its provitamin
content is a possible means for alleviating nutritional deficiencies.
beta-carotene is the main dietary precursor of vitamin A. Potato tubers contain
low levels of carotenoids, composed mainly of the xanthophylls lutein,
antheraxanthin, violaxanthin, and of xanthophyll esters. None of these
carotenoids have provitamin A activity. RESULTS: We silenced the first dedicated
step in the beta-epsilon- branch of carotenoid biosynthesis, lycopene epsilon
cyclase (LCY-e), by introducing, via Agrobacterium-mediated transformation, an
antisense fragment of this gene under the control of the patatin promoter. Real
Time measurements confirmed the tuber-specific silencing of Lcy-e. Antisense
tubers showed significant increases in beta-beta-carotenoid levels, with
beta-carotene showing the maximum increase (up to 14-fold). Total carotenoids
increased up to 2.5-fold. These changes were not accompanied by a decrease in
lutein, suggesting that LCY-e is not rate-limiting for lutein accumulation.
Tuber-specific changes in expression of several genes in the pathway were
observed. CONCLUSION: The data suggest that epsilon-cyclization of lycopene is a
key regulatory step in potato tuber carotenogenesis. Upon tuber-specific
silencing of the corresponding gene, beta-beta-carotenoid and total carotenoid
levels are increased, and expression of several other genes in the pathway is
modified.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16800876 [PubMed - indexed for MEDLINE]

1272: J Exp Bot. 2006;57(10):2445-53. Epub 2006 Jun 23.

Up- and down-regulation of Fragaria x ananassa O-methyltransferase: impacts on
furanone and phenylpropanoid metabolism.

Lunkenbein S, Salentijn EM, Coiner HA, Boone MJ, Krens FA, Schwab W.

Technical University Muenchen, Biomolecular Food Technology, Lise-Meitner-Str.
34, D-85354 Freising, Germany.

A complex mixture of hundreds of substances determines strawberry (Fragaria x
ananassa) aroma, but only approximately 15 volatiles are considered as key
flavour compounds. Of these, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) is
regarded as the most important, but it is methylated further by FaOMT (Fragaria x
ananassa O-methyltransferase) to 2,5-dimethyl-4-methoxy-3(2H)-furanone (DMMF)
during the ripening process. It is shown here that transformation of strawberry
with the FaOMT sequence in sense and antisense orientation, under the control of
the cauliflower mosaic virus 35S promoter, resulted in a near total loss of DMMF,
whereas the levels of the other volatiles remained unchanged. FaOMT repression
also affected the ratio of feruloyl 1-O-beta-D-glucose and caffeoyl
1-O-beta-D-glucose, indicating a dual function of the enzyme in planta. Thus,
FaOMT is involved in at least two different biochemical pathways in ripe
strawberry fruit.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16798852 [PubMed - indexed for MEDLINE]

1273: J Exp Bot. 2006;57(10):2363-77. Epub 2006 Jun 23.

The influence of cytosolic phosphorylating glyceraldehyde 3-phosphate
dehydrogenase (GAPC) on potato tuber metabolism.

Hajirezaei MR, Biemelt S, Peisker M, Lytovchenko A, Fernie AR, Sonnewald U.

Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3,
D-06466 Gatersleben, Germany. mohammad@ipk-gatersleben.de

The aim of this work was to investigate the importance of cytosolic
phosphorylating glyceraldehyde 3-phosphate dehydrogenase (GAPC) in potato
carbohydrate metabolism. For this purpose, the cytosolic isoform of
phosphorylating GAPC was cloned and used for an antisense approach to generate
transgenic potato plants that exhibited constitutively decreased GAPDH activity.
Potato lines with decreased activities of phosphorylating GAPC exhibited no major
changes in either whole-plant or tuber morphology. However, the levels of
3-phosphoglycerate were decreased in leaves of the transformants. A broad
metabolic phenotyping of tubers from the transformants revealed an increase in
sucrose and UDPglucose content, a decrease in the glycolytic intermediates
3-phosphoglycerate and phosphoenolpyruvate but little change in the levels of
other metabolites. Moreover, the transformants displayed no differences in cold
sweetening with respect to the wild type. Taken together these data suggest that
phosphorylating GAPC plays only a minor role in the regulation of potato
metabolism. The results presented here are discussed in relation to current
models regarding primary metabolism in the potato tuber parenchyma.

PMID: 16798850 [PubMed - indexed for MEDLINE]

1274: Microb Cell Fact. 2006 Jun 23;5:23.

Live bacterial vaccines--a review and identification of potential hazards.

Detmer A, Glenting J.

Danish Toxicology Centre, Hö¸rsholm, Denmark. ad@dhigroup.com

The use of live bacteria to induce an immune response to itself or to a carried
vaccine component is an attractive vaccine strategy. Advantages of live bacterial
vaccines include their mimicry of a natural infection, intrinsic adjuvant
properties and their possibility to be administered orally. Derivatives of
pathogenic and non-pathogenic food related bacteria are currently being evaluated
as live vaccines. However, pathogenic bacteria demands for attenuation to weaken
its virulence. The use of bacteria as vaccine delivery vehicles implies
construction of recombinant strains that contain the gene cassette encoding the
antigen. With the increased knowledge of mucosal immunity and the availability of
genetic tools for heterologous gene expression the concept of live vaccine
vehicles gains renewed interest. However, administration of live bacterial
vaccines poses some risks. In addition, vaccination using recombinant bacteria
results in the release of live recombinant organisms into nature. This places
these vaccines in the debate on application of genetically modified organisms. In
this review we give an overview of live bacterial vaccines on the market and
describe the development of new live vaccines with a focus on attenuated bacteria
and food-related lactic acid bacteria. Furthermore, we outline the safety
concerns and identify the hazards associated with live bacterial vaccines and try
to give some suggestions of what to consider during their development.

PMID: 16796731 [PubMed]

1275: Plant Cell Rep. 2006 Nov;25(11):1246-54. Epub 2006 Jun 23.

Leaf senescence is delayed in tobacco plants expressing the maize knotted1 gene
under the control of a wound-inducible promoter.

Luo K, Deng W, Xiao Y, Zheng X, Li Y, Pei Y.

Key Laboratory of Biotechnology and Crop Quality Improvement, Ministry of
Agriculture, Biotechnology Research Center, Southwest University, 400716,
Chongqing, PR China.

To extend the shelf life of freshly harvested vegetables and cut flowers, a maize
homeobox gene Knotted1 (kn1) was placed under the control of a wound-inducible
promoter win3.12 from hybrid poplar (Populus trichocarpa x P. deltoides) and
introduced into tobacco plants (Nicotiana tabacum cv. Xanthi). Transgenic
win3.12::kn1 plants were morphologically normal. A leaf-detachment assay
demonstrated that senescence in win3.12::kn1 leaves could be delayed by at least
2 weeks compared with wild type leaves. Furthermore, all leaves of win3.12::kn1
shoots remained green and healthy 3 weeks after excision and incubation in water,
while older leaves of control shoots senesced under the same conditions.
Additionally, a number of adventitious roots produced at the cut ends of wild
type shoots after a 3-week incubation, but much a less number of adventitious
roots appeared in win3.12::kn1 shoots. The delay in senescence was also confirmed
by a higher total chlorophyll (a + b) content in win3.12::kn1 leaves relative to
that of the control plants. RT-PCR analysis showed that the kn1 transcript was
detected in win3.12::kn1 leaves with wounding treatment, but otherwise was not
observed in leaves of wild type and unwounded transgenic plants. The results
presented here indicate that expression of kn1 gene driven by the wound-inducible
promoter win3.12 is potentially useful to delay senescence of vegetable crops and
commercial horticulture after harvest.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16794826 [PubMed - indexed for MEDLINE]

1276: Biotechnol Lett. 2006 Jul;28(13):959-67. Epub 2006 Jun 23.

Immunogenicity of a novel, bivalent, plant-based oral vaccine against hepatitis B
and human immunodeficiency viruses.

Shchelkunov SN, Salyaev RK, Pozdnyakov SG, Rekoslavskaya NI, Nesterov AE, Ryzhova
TS, Sumtsova VM, Pakova NV, Mishutina UO, Kopytina TV, Hammond RW.

State Research Center of Virology and Biotechnology Vector, Koltsovo, Novosibirsk
Region, 630559, Russia.

A synthetic chimeric gene, TBI-HBS, encoding the immunogenic ENV and GAG epitopes
of human immunodeficiency virus (HIV-1) and the surface protein antigen (HBsAg)
of hepatitis B virus (HBV), was expressed in tomato plants. Tomato fruits
containing the TBI-HBS antigen were fed to experimental mice and, on days 14 and
28 post-feeding, high levels of HIV- and HBV-specific antibodies were present in
the serum and feces of the test animals. Intraperitoneal injection of a DNA
vaccine directing synthesis of the same TBI-HBsAg antigen boosted the antibody
response to HIV in the blood serum; however, it had no effect on the high level
of antibodies produced to HBV.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16794774 [PubMed - indexed for MEDLINE]

1277: Biosci Biotechnol Biochem. 2006 Jun;70(6):1524-7.

Recombinant, rice-produced yeast phytase shows the ability to hydrolyze phytate
derived from seed-based feed, and extreme stability during ensilage treatment.

Hamada A, Yamaguchi K, Harada M, Horiguchi K, Takahashi T, Honda H.

Functional Chemicals Laboratory, Mitsui Chemicals, Inc., Togo, Mobara.

When fresh rice leaves producing yeast Schwanniomyces occidentalis phytase were
grounded and mixed with the whole extract of seed-based feed for pigs, the
release of orthophosphate increased significantly. More specifically, phytate, a
major source of phosphorus in the seeds, was hydrolyzed by heterologous phytase.
Moreover, when transgenic rice plants were ensiled for up to 12 weeks, no
decrease in the phytase activity of the heterologous enzyme was observed. This
result strongly suggests that transgenic rice plants producing yeast phytase can
be stored as silage without any loss of enzyme activity until usage as a feed
additive.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16794341 [PubMed - indexed for MEDLINE]

1278: Plant J. 2006 Aug;47(3):427-44. Epub 2006 Jun 22.

GAMYB controls different sets of genes and is differentially regulated by
microRNA in aleurone cells and anthers.

Tsuji H, Aya K, Ueguchi-Tanaka M, Shimada Y, Nakazono M, Watanabe R, Nishizawa
NK, Gomi K, Shimada A, Kitano H, Ashikari M, Matsuoka M.

Bioscience and Biotechnology Center, Nagoya University, Furocho, Chikusa, Nagoya
464-8601, Japan.

GAMYB is a component of gibberellin (GA) signaling in cereal aleurone cells, and
has an important role in flower development. However, it is unclear how GAMYB
function is regulated. We examined the involvement of a microRNA, miR159, in the
regulation of GAMYB expression in cereal aleurone cells and flower development.
In aleurone cells, no miR159 expression was observed with or without GA
treatment, suggesting that miR159 is not involved in the regulation of GAMYB and
GAMYB-like genes in this tissue. miR159 was expressed in tissues other than
aleurone, and miR159 over-expressors showed similar but more severe phenotypes
than the gamyb mutant. GAMYB and GAMYB-like genes are co-expressed with miR159 in
anthers, and the mRNA levels for GAMYB and GAMYB-like genes are negatively
correlated with miR159 levels during anther development. Thus, OsGAMYB and
OsGAMYB-like genes are regulated by miR159 in flowers. A microarray analysis
revealed that OsGAMYB and its upstream regulator SLR1 are involved in the
regulation of almost all GA-mediated gene expression in rice aleurone cells.
Moreover, different sets of genes are regulated by GAMYB in aleurone cells and
anthers. GAMYB binds directly to promoter regions of its target genes in anthers
as well as aleurone cells. Based on these observations, we suggest that the
regulation of GAMYB expression and GAMYB function are different in aleurone cells
and flowers in rice.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16792694 [PubMed - indexed for MEDLINE]

1279: Plant J. 2006 Aug;47(3):457-66. Epub 2006 Jun 22.

Cloning of two splice variants of the rice PTS1 receptor, OsPex5pL and OsPex5pS,
and their functional characterization using pex5-deficient yeast and Arabidopsis.

Lee JR, Jang HH, Park JH, Jung JH, Lee SS, Park SK, Chi YH, Moon JC, Lee YM, Kim
SY, Kim JY, Yun DJ, Cho MJ, Lee KO, Lee SY.

Environmental Biotechnology National Core Research Center, Gyeongsang National
University, Jinju 660-701, Korea.

Using the rice PEX14 cDNA as a bait in a yeast two-hybrid assay, two splice
variants of the type I peroxisomal targeting signal (PTS1) receptor, OsPex5pL and
OsPex5pS, were cloned from a pathogen-treated rice leaf cDNA library. The
proteins were produced from a single gene by alternative splicing, which
generated a full-length variant, OsPEX5L, and a variant that lacked exon 7,
OsPEX5S. OsPex5pL contained 11 copies of the pentapeptide motif WXXXF/Y in its
N-terminus, and seven tetratricopeptide repeats in its C-terminus. Expression of
OsPEX5L and OsPEX5S predominantly occurred in leaf tissues, and was induced by
various stresses, such as exposure to the pathogen Magnaporthe grisea, and
treatment with fungal elicitor, methyl viologen, NaCl or hydrogen peroxide. The
Arabidopsis T-DNA insertional pex5 mutant, Atpex5, which does not germinate in
the absence of sucrose and was resistant to indole-3-butyric acid (IBA), was
perfectly rescued by over-expression of OsPex5pL, but not by OsPex5pS. Using
transient expression of OsPex5pL and OsPex5pS in the Atpex5 mutant, we show that
OsPex5pL translocates both PTS1- and PTS2-containing proteins into the peroxisome
by interacting with OsPex7p, whereas OsPex5pS is involved only in PTS1-dependent
import in Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16792693 [PubMed - indexed for MEDLINE]

1280: Ann Bot (Lond). 2006 Sep;98(3):565-71. Epub 2006 Jun 21.

Accumulation of glycinebetaine in rice plants that overexpress choline
monooxygenase from spinach and evaluation of their tolerance to abiotic stress.

Shirasawa K, Takabe T, Takabe T, Kishitani S.

Graduate School of Agricultural Science, Tohoku University, Aoba, Sendai
981-8555, Japan.

BACKGROUND AND AIMS: Glycinebetaine (GB), a quaternary ammonium compound, is a
very effective compatible solute. In higher plants, GB is synthesized from
choline (Cho) via betaine aldehyde (BA). The first and second steps in the
biosynthesis of GB are catalysed by choline monooxygenase (CMO) and by betaine
aldehyde dehydrogenase (BADH), respectively. Rice (Oryza sativa), which has two
genes for BADH, does not accumulate GB because it lacks a functional gene for
CMO. Rice plants accumulate GB in the presence of exogenously applied BA, which
leads to the development of a significant tolerance to salt, cold and heat
stress. The goal in this study was to evaluate and to discuss the effects of
endogenously accumulated GB in rice. METHODS: Transgenic rice plants that
overexpressed a gene for CMO from spinach (Spinacia oleracea) were produced by
Agrobacterium-mediated transformation. After Southern and western blotting
analysis, GB in rice leaves was quantified by (1)H-NMR spectroscopy and the
tolerance of GB-accumulating plants to abiotic stress was investigated. KEY
RESULTS: Transgenic plants that had a single copy of the transgene and expressed
spinach CMO accumulated GB at the level of 0.29-0.43 micromol g(-1) d. wt and had
enhanced tolerance to salt stress and temperature stress in the seedling stage.
CONCLUSIONS: In the CMO-expressing rice plants, the localization of spinach CMO
and of endogenous BADHs might be different and/or the catalytic activity of
spinach CMO in rice plants might be lower than it is in spinach. These
possibilities might explain the low levels of GB in the transgenic rice plants.
It was concluded that CMO-expressing rice plants were not effective for
accumulation of GB and improvement of productivity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16790464 [PubMed - indexed for MEDLINE]

1281: J Agric Food Chem. 2006 Jun 28;54(13):4633-40.

Piceid (resveratrol glucoside) synthesis in stilbene synthase transgenic apple
fruit.

Rö¼hmann S, Treutter D, Fritsche S, Briviba K, Szankowski I.

Institute for Biological Production Systems, Fruit Science Section, University of
Hanover, Herrenhaeuser Strasse 2, 30419 Hannover, Germany.

A stilbene synthase gene along with the selectable marker gene bar for herbicide
resistance was transferred via Agrobacterium tumefaciens mediated transformation
into apple (Malus domesticaBorkh.) cvs. 'Elstar' and 'Holsteiner Cox'. The
stilbene synthase catalyzes the conversion of 1 molecule of p-coumaroyl-CoA and 3
molecules of malonyl-CoA into 3,4',5-trihydroxystilbene, commonly known as
resveratrol. This phytoalexin has implications in both phytopathology and human
health. Greenhouse-grown transgenic and nontransformed control plants were
grafted onto dwarfing rootstock M27. Flowering and fruiting occurred within the
following years, offering the opportunity to analyze transgenic apple fruit and
fertility of transgenic plants as well as inheritance of the transgenes into the
seedling progeny. Molecular analysis revealed that the stilbene synthase is
expressed in transgenic plants and in the skin and flesh of transgenic apple
fruit. After formation, resveratrol is modified by the addition of a hexose
sugar. The resulting component was characterized as piceid. With the aim of
characterizing the influence of the novel biosynthetic pathway on the
accumulation of other phenolic compounds naturally present in apple fruit, the
amounts of flavanols, flavonols, phloretin derivatives and hydroxycinnamic acids
in wild type and transgenic fruit were determined by HPLC. In all investigated
transformed lines that accumulated piceid, no negative correlation between levels
of piceid and the above-mentioned compounds was observed, except for the flavonol
contents, which slightly decreased. Inheritance of the transgenes was confirmed
in the seedling progeny, which were obtained after pollination of transgenic
plants with nontransgenic pollen and vice versa after pollination of
nontransgenic plants with pollen obtained from transgenic plants. The fertility
of stilbene synthase transgenic plants was demonstrated. To the authors'
knowledge this is the first time that data are available on piceid synthesis in
transgenic apple fruit and the effects of its accumulation on levels of other
phenolic compounds present in the fruit.

PMID: 16787008 [PubMed - indexed for MEDLINE]

1282: J Agric Food Chem. 2006 Jun 28;54(13):4624-32.

Heat-stable phytases in transgenic wheat (Triticum aestivum L.): deposition
pattern, thermostability, and phytate hydrolysis.

Brinch-Pedersen H, Hatzack F, Stö¶ger E, Arcalis E, Pontopidan K, Holm PB.

Research Centre Flakkebjerg, Department of Genetics and Biotechnology, Danish
Institute of Agricultural Sciences, DK-4200 Slagelse, Denmark.
Henrik.brinchpedersen@agrsci.dk

The present paper addresses the question of thermotolerance of in planta
synthesized heterologous enzymes using phytase as a model. Two individual
transgenic wheat materials expressing an Aspergillus fumigatus phytase with a low
denaturation temperature (62.5 degrees C) but a high refolding capacity, and a
rationally designed consensus phytase engineered to a high denaturation
temperature (89.3 degrees C), were evaluated. High levels of endosperm specific
expression were ensured by the wheat high molecular weight glutenin 1DX5
promoter. Immunodetection at the light and electron microscopical level shows
unequivocally that the heterologous phytase is deposited in the vacuole, albeit
that the transformation constructs were designed for secretion to the apoplast.
Evaluation of heat stability properties and kinetic properties unraveled that,
under these deposition conditions, heat stability based on high unfolding
temperature is superior to high refolding capacity and represents a realistic
strategy for improving phosphate and mineral bioavailability in cereal-based feed
and food.

PMID: 16787007 [PubMed - indexed for MEDLINE]

1283: Food Nutr Bull. 2006 Jun;27(2):167-79.

Agricultural biodiversity, nutrition, and health: making a difference to hunger
and nutrition in the developing world.

Frison EA, Smith IF, Johns T, Cherfas J, Eyzaguirre PB.

International Plant Genetic Resources Institute, Rome, Italy. e.frison@cgiar.org

BACKGROUND: In spite of the strides made globally in reducing hunger, the
problems of micronutrient deficiencies and coexisting obesity and related
cardiovascular and degenerative diseases constitute a formidable challenge for
the future. Attempts to reverse this trend with single-nutrient intervention
strategies have met with limited success, resulting in renewed calls for
food-based approaches. The deployment of agricultural biodiversity is an approach
that entails greater use of local biodiversity to ensure dietary diversity.
OBJECTIVE: To outline a new strategy proposed by the International Plant Genetic
Resources Institute (IPGRI) that employs agricultural biodiversity as the primary
resource for food security and health. METHODS: The authors carried out a
meta-analysis to review and assemble existing information on the nutritional and
healthful properties of traditional foods based on a diverse set of case studies
and food composition and nutritional analysis studies. The methods highlight
particular examples of foods where analysis of nutrient and non-nutrient
composition reveals important traits to address the growing problems of
malnutrition associated with the rise of chronic diseases. Finally, the authors
analyze social, economic, and cultural changes that undermine the healthful
components of traditional diets. RESULTS: Based on this multidisciplinary and
comparative approach, the authors suggest a holistic food-based approach that
combines research to assess and document nutritional and healthful properties of
traditional foods, investigating options in which nutritionally valuable
traditional foods can contribute to better livelihoods, and ways that awareness
and promotional campaigns can identify healthful components of traditional diets
that fit the needs of urban and market-oriented consumers. CONCLUSIONS: There is
an urgent need for agricultural research centers, national agricultural research
systems, universities, and community-based organizations to work together under a
shared policy framework with the aim of developing a strong evidence base linking
biodiversity, nutrition, and health. Although these initiatives are still
ongoing, the gains realized in small-scale and local pilot efforts have
encouraged IPGRI to work with local partners toward the implementation of
scale-up efforts in various regions.

Publication Types:
Meta-Analysis
Review

PMID: 16786983 [PubMed - indexed for MEDLINE]

1284: Plant Cell Rep. 2006 Nov;25(11):1181-92. Epub 2006 Jun 20.

Expression of ipt gene controlled by an ethylene and auxin responsive fragment of
the LEACO1 promoter increases flower number in transgenic Nicotiana tabacum.

Khodakovskaya M, Zhao D, Smith W, Li Y, McAvoy R.

Plant Science Department, University of Connecticut, 06269-4163, Storrs, CT, USA.

Cytokinins play important roles in regulating plant growth and development. A new
genetic construct for regulating cytokinin content in plant cells was cloned and
tested. The gene coding for isopentenyl transferase (ipt) was placed under the
control of a 0.821 kb fragment of the 1-aminocyclopropane-1-carboxylic acid (ACC)
oxidase gene promoter from Lycopersicon esculentum (LEACO1) and introduced into
Nicotiana tabacum (cv. Havana). Some LEACO1(0.821) (kb)-ipt transgenic plant
lines displayed normal shoot morphology but with a dramatic increase in the
number of flower buds compared to nontransgenic plants. Other transgenic lines
produced excessive lateral branch development but no change in flower bud number.
Isolated leaves of transgenic tobacco plants showed a significantly prolonged
retention of chlorophyll under dark incubation (25 degrees C for 20 days). Leaves
of nontransformed plants senesced gradually under the same conditions.
Experiments with LEACO1(0.821) (kb)-gus transgenic tobacco plants suggested auxin
and ethylene involvement in induction of LEACO1(0.821) (kb) promoter activity.
Multiple copies of nucleotide base sequences associated with either ethylene or
auxin response elements were identified in the LEACO1(0.821) (kb) promoter
fragment. The LEACO1(0.821) (kb)-ipt fusion gene appears to have potential
utility for improving certain ornamental and agricultural crop species by
increasing flower bud initiation and altering branching habit.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16786314 [PubMed - indexed for MEDLINE]

1285: Biotechnol Lett. 2006 Jun;28(12):869-75. Epub 2006 May 31.

High-level expression of basic fibroblast growth factor in transgenic soybean
seeds and characterization of its biological activity.

Ding SH, Huang LY, Wang YD, Sun HC, Xiang ZH.

Lab of Biotechnology, Collage of Fishery Science, Southwest University,
Chongqing, China.

The glycinin G1 gene encodes a soybean seed storage protein accumulating at a
high level. We have used the G1 promoter to confer seed-specific expression of
human basic fibroblast growth factor (bFGF) in transgenic soybeans. The coding
region of 18 kDa bFGF was fused to the promoter or promoter-signal peptide
sequence of G1 gene, and transferred into soybean. Analysis of transgenic plants
demonstrated that bFGF transcript or protein was confined to the seeds. The
highest level of bFGF accumulation in the seeds reached up to 2.3% of total
soluble protein. The soybean-derived bFGF was biologically active as confirmed by
its mitogenic activity on Balb/c 3T3 cells, and exhibited other properties
identical to native bFGF. We also observed a seed-specific expression of
beta-glucuronidase driven by the G1 promoter. These results indicated that the G1
promoter contains essential cis-elements for seed-specific expression, and thus
can be used for expression of pharmaceutical proteins in soybean seeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16786271 [PubMed - indexed for MEDLINE]

1286: Theor Appl Genet. 2006 Jun;113(1):128-36. Epub 2006 Apr 25.

Comparative field performance over 3 years and two sites of transgenic wheat
lines expressing HMW subunit transgenes.

Shewry PR, Powers S, Field JM, Fido RJ, Jones HD, Arnold GM, West J, Lazzeri PA,
Barcelo P, Barro F, Tatham AS, Bekes F, Butow B, Darlington H.

Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK.
peter.shewry@bbsrc.ac.uk

A series of transgenic wheat lines expressing additional high molecular weight
(HMW) subunit genes and the corresponding control lines were grown in replicate
field trials at two UK sites (Rothamsted Research, approximately 50 km north of
London and Long Ashton, near Bristol) over 3 years (1998, 1999, 2000), with
successive generations of the transgenic lines (T3, T4, T5) being planted. Four
plots from each site were used to determine grain dry weight, grain nitrogen,
dough strength (measured as peak resistance by Mixograph analysis) and the
expression levels of the endogenous and "added" subunits. Detailed statistical
analyses showed that the transgenic and non-transgenic lines did not differ in
terms of stability of HMW subunit gene expression or in stability of grain
nitrogen, dry weight or dough strength, either between the 3 years or between
sites and plots. These results indicate that the transgenic and control lines can
be regarded as substantially equivalent in terms of stability of gene expression
between generations and environments.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16783593 [PubMed - indexed for MEDLINE]

1287: Transgenic Res. 2006 Jun;15(3):313-24.

Rust and downy mildew resistance in pearl millet (Pennisetum glaucum) mediated by
heterologous expression of the afp gene from Aspergillus giganteus.

Girgi M, Breese WA, Lö¶rz H, Oldach KH.

Developmental Biology and Biotechnology, Biocenter Klein Flottbek, University of
Hamburg, Ohnhorststr. 18, 22609 Hamburg, Germany. girgi@botanik.uni-hamburg.de

The cDNA encoding the antifungal protein AFP from the mould Aspergillus giganteus
was introduced into two pearl millet (Pennisetum glaucum) genotypes by particle
bombardment. Stable integration and expression of the afp gene was confirmed in
two independent transgenic T0 plants and their progeny using Southern blot and
RT-PCR analysis. In vitro infection of detached leaves and in vivo inoculation of
whole plants with the basidomycete Puccinia substriata, the causal agent of rust
disease, and the oomycete Sclerospora graminicola, causal agent of downy mildew,
resulted in a significant reduction of disease symptoms in comparison to wild
type control plants. The disease resistance of pearl millet was increased by up
to 90% when infected with two diverse, economically important pathogens. This is
the first report of genetic enhancement of Pennisetum glaucum against fungal
infections.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16779647 [PubMed - indexed for MEDLINE]

1288: Transgenic Res. 2006 Jun;15(3):305-11.

Stability of soybean recombinant plastome over six generations.

Dufourmantel N, Tissot G, Garö§on F, Pelissier B, Dubald M.

BioScience, Bayer CropScience, 14-20 rue Pierre Baizet, 69263 Lyon, France.

The stability of a plastid transgene has been evaluated in soybean transformants
over six generations. These transformants had integrated the aadA selection
cassette in the intergenic region between the rps12/7 and trnV genes. Three
independent homoplasmic T0 transformation events were selected and ten plants
from each event propagated to generation T5 in the absence of selection pressure.
No transgene rearrangement nor wild-type plastome were detected in generation T5
by Southern blot analysis. All tested progenies were uniformly resistant to
spectinomycin. Therefore, soybean transformants of generations T0 and T5 appear
to be genetically and phenotypically identical.

PMID: 16779646 [PubMed - indexed for MEDLINE]

1289: Transgenic Res. 2006 Jun;15(3):291-304.

Intron-hairpin RNA derived from replication associated protein C1 gene confers
immunity to tomato yellow leaf curl virus infection in transgenic tomato plants.

Fuentes A, Ramos PL, Fiallo E, Callard D, Sö¡nchez Y, Peral R, Rodrö­guez R, Pujol
M.

Plant Virology Laboratory, Plant Department, Center for Genetic Engineering and
Biotechnology, P.O. Box 6162, 10600, Havana, Cuba.

The whitefly-transmitted Tomato Yellow Leaf Curl Virus (TYLCV) is the major
pathogen of tomato crop in Cuba and one of the most outstanding viral diseases of
plants worldwide. In this work, we have developed transgenic tomato plants,
transformed with an intron-hairpin genetic construction to induce post-
transcriptional gene silencing against the early TYLCV replication associated
protein gene (C1). The intron-hairpin RNA produced involves 726 nts of the 3' end
of the TYLCV C1 gene as the arms of the hairpin, and the castor bean catalase
intron. Transgenic tomato plants belonging to line 126, which harbor a single
transgene copy, showed immunity to TYLCV, even in extreme conditions of infection
(4-leaf-stage plants and 300 to many hundreds viruliferous whiteflies per plant
during 60 days). Dot blot hybridization of these plants showed no TYLCV DNA
presence 60 days after inoculation. Small interfering RNA molecules were detected
in both inoculated and non-inoculated plants from line 126. These transgenic
tomato plants of the otherwise very TYLCV-susceptible Campbell-28 tomato
cultivar, are the first report of resistance to a plant DNA virus obtained by the
use of the intron-hairpin RNA approach.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16779645 [PubMed - indexed for MEDLINE]

1290: Transgenic Res. 2006 Jun;15(3):277-89.

Mycotoxin reduction in Bt corn: potential economic, health, and regulatory
impacts.

Wu F.

Environmental, Occupational Health, Graduate School of Public Health, University
of Pittsburgh, 130 DeSoto St., Pittsburgh, PA 15261, USA. fwu@eoh.pitt.edu

Genetically modified (GM) Bt corn, through the pest protection that it confers,
has lower levels of mycotoxins: toxic and carcinogenic chemicals produced as
secondary metabolites of fungi that colonize crops. In some cases, the reduction
of mycotoxins afforded by Bt corn is significant enough to have an economic
impact, both in terms of domestic markets and international trade. In less
developed countries where certain mycotoxins are significant contaminants of
food, Bt corn adoption, by virtue of its mycotoxin reduction, may even improve
human and animal health. This paper describes an integrated assessment model that
analyzes the economic and health impacts of two mycotoxins in corn: fumonisin and
aflatoxin. It was found that excessively strict standards of these two mycotoxins
could result in global trade losses in the hundreds of millions US dollars
annually, with the US, China, and Argentina suffering the greatest losses. The
paper then discusses the evidence for Bt corn's lower levels of contamination of
fumonisin and aflatoxin, and estimates economic impacts in the United States. A
total benefit of Bt corn's reduction of fumonisin and aflatoxin in the US was
estimated at 23 million dollars annually. Finally, the paper examines the
potential policy impacts of Bt corn's mycotoxin reduction, on nations that are
making a decision on whether to allow commercialization of this genetically
modified crop.

PMID: 16779644 [PubMed - indexed for MEDLINE]

1291: Science. 2006 Jun 16;312(5780):1586-7.

Agriculture. A kinder, gentler Jeremy Rifkin endorses biotech, or does he?

Stokstad E.

Publication Types:
Biography
Historical Article
News

Personal Name as Subject:
Rifkin J

PMID: 16778031 [PubMed - indexed for MEDLINE]

1292: Plant Physiol. 2006 Aug;141(4):1376-88. Epub 2006 Jun 15.

A novel nuclear-localized CCCH-type zinc finger protein, OsDOS, is involved in
delaying leaf senescence in rice.

Kong Z, Li M, Yang W, Xu W, Xue Y.

Laboratory of Molecular and Developmental Biology, Institute of Genetics and
Developmental Biology, Chinese Academy of Sciences and National Centre for Plant
Gene Research, Beijing 100080, China.

Leaf senescence is a developmentally programmed degeneration process, which is
fine tuned by a complex regulatory network for plant fitness. However, molecular
regulation of leaf senescence is poorly understood, especially in rice (Oryza
sativa), an important staple crop for more than half of the world population.
Here, we report a novel nuclear-localized CCCH-type zinc finger protein, Oryza
sativa delay of the onset of senescence (OsDOS), involved in delaying leaf
senescence in rice. The expression of OsDOS was down-regulated during natural
leaf senescence, panicle development, and pollination, although its transcripts
were accumulated in various organs. RNAi knockdown of OsDOS caused an accelerated
age-dependent leaf senescence, whereas its overexpression produced a marked delay
of leaf senescence, suggesting that it acts as a negative regulator for leaf
senescence. A genome-wide expression analysis further confirmed its negative
regulation for leaf senescence and revealed that, in particular, the jasmonate
(JA) pathway was found to be hyperactive in the OsDOS RNAi transgenic lines but
impaired in the OsDOS overexpressing transgenic lines, indicating that this
pathway is likely involved in the OsDOS-mediated delaying of leaf senescence.
Furthermore, methyl JA treatments of both seeds and detached leaves from the RNAi
and the overexpressing transgenic lines showed hyper- and hyporesponses,
respectively, consistent with the negative regulation of the JA pathway by OsDOS.
Together, these results indicate that OsDOS is a novel nuclear protein that
delays leaf senescence likely, at least in part, by integrating developmental
cues to the JA pathway.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16778011 [PubMed - indexed for MEDLINE]

1293: J Plant Physiol. 2006 Jul;163(8):856-62. Epub 2005 Oct 25.

Plant-specific insertions in the soybean aspartic proteinases, soyAP1 and soyAP2,
perform different functions of vacuolar targeting.

Terauchi K, Asakura T, Ueda H, Tamura T, Tamura K, Matsumoto I, Misaka T,
Hara-Nishimura I, Abe K.

Department of Applied Biological Chemistry, Graduate School of Agricultural and
Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.

Most aspartic proteinases (APs) of plant origin are characterized by the presence
of plant-specific insertion (PSI) in their primary structure. PSI has been
reported to function as signals for both transport of AP molecules from the
endoplasmic reticulum (ER) and for their targeting to the vacuole. To determine
the functions of the PSIs in soyAP1 and soyAP2 identified in our previous study,
we examined their subcellular localization by transient expression of a green
fluorescent protein (GFP) fusion protein in the protoplasts of Arabidopsis
suspension-cultured cells. Both soyAP1-GFP and soyAP2-GFP were targeted to the
vacuole. To confirm the role of the PSI, we prepared PSI-deleted soyAP1 and
soyAP2, and investigated their vacuolar targeting by the same method. While the
former deletion mutant was always transported to the vacuole, the latter
sometimes remained in the ER and was only sometimes transported to the vacuole.
These observations indicated that, in the case of soyAP1, the PSI is not involved
in vacuolar targeting, also suggesting that the function of the PSI differs
depending on its origin.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16777533 [PubMed - indexed for MEDLINE]

1294: Mol Plant Microbe Interact. 2006 Jun;19(6):655-64.

Mi-1-Mediated aphid resistance involves salicylic acid and mitogen-activated
protein kinase signaling cascades.

Li Q, Xie QG, Smith-Becker J, Navarre DA, Kaloshian I.

Department of Nematology, University of California, Riverside, CA 92521, USA.

The tomato Mi-1 gene confers resistance to root-knot nematodes (Meloidogyne
spp.), potato aphids (Macrosiphum eluphorbiae), and whiteflies (Bemisia tabaci
and B. tabaci biotype B). Resistance to potato aphid is developmentally regulated
and is not associated with induction of a hypersensitive response. The NahG
transgene that eliminates endogenous salicylic acid (SA) was used to test the
role of the SA signaling pathway in the resistance mediated by Mi-1 to potato
aphids. Aphids survived longer on NahG tomato plants than on wild type. However,
aphid reproduction was not affected on NahG tomato. Aphid resistance in Mi-1 NahG
plants was completely abolished and the phenotype was successfully rescued by
application of BTH (benzo(1,2,3)-thiaiazole-7-carbothioic acid S-methyl ester),
indicating that the SA signaling pathway is an important component of
Mi-1-mediated aphid resistance. Using virus-induced gene silencing, one or more
mitogen-activated protein kinase (MAPK) cascades required for Mi-1-mediated aphid
resistance were identified. Silencing plants for MAPK kinase (LeMKK2) and MAPKs
(LeMPK2 and LeMPK1, or LeMPK3) resulted in attenuation of Mi-1-mediated aphid
resistance. These results further demonstrate that resistance gene-mediated
signaling events against piercing-sucking insects are similar to those against
other plant pathogens.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16776299 [PubMed - indexed for MEDLINE]

1295: Mol Plant Microbe Interact. 2006 Jun;19(6):614-24.

Analysis of silencing escape of tomato leaf curl virus: an evaluation of the role
of DNA methylation.

Bian XY, Rasheed MS, Seemanpillai MJ, Ali Rezaian M.

CSIRO Plant Industry, PO Box 350, Glen Osmond, SA 5064, Australia.

RNA silencing is a sequence-specific mechanism regulating gene expression and has
been used successfully for antiviral defense against RNA viruses. Similar
strategies to develop resistance against DNA containing Tomato leaf curl virus
(TLCV) and some other geminiviruses have been unsuccessful. To analyze this
silencing escape, we transformed tomato plants with a hairpin construct from the
TLCV C2 open reading frame (ORF). The transgenic plants showed a strong RNA
silencing response, and following TLCV inoculation, their infection was delayed.
However, the viral infection was not prevented and TLCV DNA accumulated to the
levels found in nontransgenic plants. To determine the fate of a transgene
carrying homology to the virus, we used transgenic plants carrying the TLCV C4
gene, which induces a distinct phenotype. Upon TLCV infection, the phenotype was
abolished and C4 transcript disappeared. Concurrently, TLCV-specific small
interfering RNAs were produced. In situ hybridization showed abundant levels of
TLCV DNA in phloem cells of TLCV-infected C4 transgenic plants. However, the C4
transcripts were no longer detectable in nonvascular cells. Analysis of the
transgene by methylation sequencing revealed a high level of de novo methylation
of asymmetric cytosines in both the C4 ORF and its 35S promoter. A high level of
methylation also was found at both symmetric and asymmetric cytosines of the
complementary-sense strand of TLCV double-stranded DNA. Given the previous
finding that methylated geminiviral DNA is not competent for replication, we
provide a model whereby TLCV evades host defense through a population of de novo
synthesized unmethylated DNA.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16776295 [PubMed - indexed for MEDLINE]

1296: Mol Plant Microbe Interact. 2006 Jun;19(6):567-76.

CDNA-AFLP combined with functional analysis reveals novel genes involved in the
hypersensitive response.

Gabriö«ls SH, Takken FL, Vossen JH, de Jong CF, Liu Q, Turk SC, Wachowski LK,
Peters J, Witsenboer HM, de Wit PJ, Joosten MH.

Laboratory of Phytopathology, Wageningen University, Binnenhaven 5, 6709 PD
Wageningen, The Netherlands.

To identify genes required for the hypersensitive response (HR), we performed
expression profiling of tomato plants mounting a synchronized HR, followed by
functional analysis of differentially expressed genes. By cDNA-AFLP analysis, the
expression profile of tomato plants containing both the Cf-4 resistance gene
against Cladosporium fulvum and the matching Avr4 avirulence gene of this fungus
was compared with that of control plants. About 1% of the transcript-derived
fragments (442 out of 50,000) were derived from a differentially expressed gene.
Based on their sequence and expression, 192 fragments, referred to as
Avr4-responsive tomato (ART) fragments, were selected for VIGS (virus-induced
gene silencing) in Cf-4-transgenic Nicotiana benthamiana. Inoculated plants were
analyzed for compromised HR by agroinfiltration of either the C. fulvum Avr4 gene
or the Inf1 gene of Phytophthora infestans, which invokes a HR in wild-type N.
benthamiana. VIGS using 15 of the ART fragments resulted in a compromised HR,
whereas VIGS with fragments of ART genes encoding HSP90, a nuclear GTPase, an L19
ribosomal protein, and most interestingly, a nucleotide binding-leucine rich
repeat (NB-LRR)-type protein severely suppressed the HR induced both by Avr4 and
Inf1. Requirement of an NB-LRR protein (designated NRC1, for NB-LRR protein
required for HR-associated cell death 1) for Cf resistance protein function as
well as Inf1-mediated HR suggests a convergence of signaling pathways and
supports the recent observation that NB-LRR proteins play a role in signal
transduction cascades downstream of resistance proteins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16776290 [PubMed - indexed for MEDLINE]

1297: Plant Cell Rep. 2006 Nov;25(11):1193-8. Epub 2006 Jun 15.

Expression of recombinant Digitalis lanata EHRH. cardenolide 16'-O-glucohydrolase
in Cucumis sativus L. hairy roots.

Shi HP, Lindemann P.

Institut fö¼r Pharmakologie und Pharmazeutische Biologie,
Martin-Luther--Universitö¤t, Hoher Weg 8, 06120, Halle (Saale), Germany.

The coding sequence for the Digitalis lanata EHRH. cardenolide
16'-O-glucohydrolase was inserted downstream of the 35S promoter in the binary
vector pBI121 resulting in plant expression vector pBI121cgh. Cotyledon explants
excised from 10-day-old seedlings of Cucumis sativus L. were transformed using
Agrobacterium rhizogenes 15834 harbouring pBI121cgh. Hairy roots were obtained
from infected cotyledon explants in vitro 10 days after inoculation. PCR
amplification of coding sequences for cgh I, rolB and rolC from Ri plasmid showed
that the aimed sequences were inserted into the genome of transformed cucumber
hairy roots. Glycolytic activity of the transgenic CGH I was measured by HPLC
using Lanatoside glycosides as substrate. Therefore, the cgh I transformed
cucumber hairy roots may provide a valuable model for biotransformation of
natural compounds by recombinant enzymes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16775721 [PubMed - indexed for MEDLINE]

1298: Plant J. 2006 Aug;47(3):417-26. Epub 2006 Jun 15.

Mutations in the eIF(iso)4G translation initiation factor confer high resistance
of rice to Rice yellow mottle virus.

Albar L, Bangratz-Reyser M, Hö©brard E, Ndjiondjop MN, Jones M, Ghesquiö¨re A.

UMR 5096, IRD/CNRS/Universitö© de Perpignan, BP 64501, 34394 Montpellier CEDEX 5,
France. laurence.albar@mpl.ird.fr

We report here evidence of the role that the isoform of the eukaryotic
translation initiation factor 4G (eIF(iso)4G) plays in naturally occurring
resistance in plant/virus interactions. A genetic and physical mapping approach
was developed to isolate the Rymv1 locus controlling the high recessive
resistance to Rice yellow mottle virus (RYMV) in the rice (Oryza sativa) variety
Gigante. The locus was mapped to a 160-kb interval containing a gene from the
eIF(iso)4G family. The stable transformation of a resistant line with the cDNA of
this gene, derived from a susceptible variety, resulted in the loss of resistance
in transgenic plants. The allelic variability of this gene was analysed in three
resistant and 17 susceptible varieties from different cultivated rice species or
subspecies. Compared with susceptible varieties, resistant varieties present
specific alleles, characterized by either amino acid substitutions or short
amino-acid deletions in the middle domain of the protein. The structure of this
domain was modelled and showed that the substitutions were clustered on a small
surface patch. This suggests that this domain may be involved in an interaction
with the virus.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16774645 [PubMed - indexed for MEDLINE]

1299: J Zhejiang Univ Sci B. 2006 Jul;7(7):591-5.

Erratum in:
J Zhejiang Univ Sci B. 2007 Feb;8(2):115.

Characteristics of transgenic tomatoes antisensed for the ethylene receptor genes
LeETR1 [corrected] and LeETR2 [corrected]

Wang ZF, Ying TJ, Zhang Y, Bao BL, Huang XD.

School of Biosystem Engineering and Food Science, Zhejiang University, Hangzhou
310029, China.

Two stable transformed lines containing antisense LeETR1 [corrected] or LeETR2
[corrected] sequences and their hybridized line were investigated to determine
the effect of LeETR1 [corrected] and LeETR2 [corrected] specificity in the
ethylene receptor family in tomato (Lycopersicon esculentum Mill.) on ethylene
signaling. The transgenic line ale1 containing antisense LeETR1 [corrected]
displayed shorter length of seedling grown in the dark and adult plant in the
light, severe epinastic petiole, and accelerated abscission of petiole explant
and senescence of flower explant, compared with its wild type B1. The transgenic
line ale2 containing antisense LeETR2 [corrected] also exhibited shorter
hypocotyls and slightly accelerated abscission. The phenotypes of cross line dale
of LeETR1 [corrected] and LeETR2 [corrected] were close to ale1 in many aspects.
These results suggested that LeETR1 [corrected] probably plays a relatively
important role in ethylene signaling of tomato growth and development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16773735 [PubMed - indexed for MEDLINE]

1300: Plant Cell Rep. 2006 Nov;25(11):1199-204. Epub 2006 Jun 14.

Transgenic wheat progeny resistant to powdery mildew generated by Agrobacterium
inoculum to the basal portion of wheat seedling.

Zhao TJ, Zhao SY, Chen HM, Zhao QZ, Hu ZM, Hou BK, Xia GM.

School of Life Sciences, Shandong University, Jinan, 250100, PR China.

To improve the transformation efficiency of wheat (Triticum aestivum L.) mediated
by Agrobacterium tumefaciens, we explored the possibility of employing the basal
portion of wheat seedling (shoot apical meristem) as the explants. Three
genotypes of wheat were transformed by A. tumefaciens carrying beta-1,
3-glucanase gene. After vernalization, the seeds to be transformed were
germinated. When these seedlings grew up to 2 approximately 5 cm, their
coleoptile and half of the cotyledon were cut out, and the basal portions were
infected by A. tumefaciens. A total 27 T(0) transgenic plants were obtained, and
the average transformation efficiency was as high as 9.82%. Evident segregation
occurred in some of the T(1) plants, as was indicated by PCR and Southern
blotting analysis. Investigation of the T(2) plants revealed that some
transformed plants had higher resistance to powdery mildew than the controls.
Northern blotting revealed that beta-1, 3-glucanase gene was normally expressed
in the T(2) plants, which showed an increased resistance to powdery mildew. The
results above indicate that the exogenous gene has been successfully integrated
into the genome of wheat, transmitted and expressed in the transgenic progeny.
From all the results above, it can be concluded that Agrobacterium inoculum to
the basal portion of wheat seedling is a highly efficient and dependable
transformation method. It can be developed into a practicable method for transfer
of target gene into wheat.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16773333 [PubMed - indexed for MEDLINE]

1301: Arch Virol. 2006 Nov;151(11):2111-22. Epub 2006 Jun 19.

Expression of rice yellow mottle virus coat protein enhances virus infection in
transgenic plants.

Kouassi NK, Chen L, Sirö© C, Bangratz-Reyser M, Beachy RN, Fauquet CM, Brugidou C.

Centre National de Recherche Agronomique (CNRA), Laboratoire Central de
Biotechnologies, Abidjan, Cö´te d'Ivoire.

The disease caused by rice yellow mottle virus (RYMV) is a major, economically
important constraint to rice production in Africa. RYMV is mechanically
transmitted by a variety of agents, including insect vectors. The production of
resistant rice varieties would be an important advance in the control of the
disease and increase rice production in Africa. We produced transgenic plants of
the Oryza sativa japonica variety, TP309, to express a RYMV coat protein gene
(CP) and mutants of the CP under the control of a ubiquitin promoter. Transgenic
plants expressing genes that encode wild-type CP (wt.CP), deleted CP
(DeltaNLS.CP), mRNA of the CP, or antisense CP sequences of the CP gene were
characterised. Eighty per cent (80%) of independent transgenic lines analysed
contained CP gene sequences. Transgenic plants were challenged with RYMV and
produced two types of reactions. Most of the plants expressing antisense
sequences of the CP and untranslatable CP mRNA exhibited a delay in virus
accumulation of up to a week, and the level of virus accumulation was reduced
compared with non-transgenic TP309 plants. Transgenic plants expressing RYMV
wild-type CP (wt.CP) and deleted CP (DeltaNLS.CP) accumulated the highest levels
of virus particles. These results suggest that antisense CP and untranslatable CP
mRNA induced moderate resistance, whereas transgenic CP enhanced virus infection.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16773235 [PubMed - indexed for MEDLINE]

1302: Plant Cell Rep. 2006 Nov;25(11):1157-65. Epub 2006 Jun 13.

Isolation of a maize beta-glucosidase gene promoter and characterization of its
activity in transgenic tobacco.

Gu R, Zhao L, Zhang Y, Chen X, Bao J, Zhao J, Wang Z, Fu J, Liu T, Wang J, Wang
G.

State Key Laboratory for Agrobiotechnology and National Center for Maize
Improvement, China Agricultural University, Beijing, 100094, China.

The beta-glucosidase gene of maize (ZmGLU1) was suggested to hydrolyze
cytokinin-conjugate and release free cytokinin during plant growth and
development. A clone containing the upstream region of ZmGLU1 was isolated and
sequenced from a maize genomic library. The full-length ZmGLU1 promoter and a
series of its 5' deletions were fused to the beta-glucuronidase (GUS) reporter
gene and transferred into tobacco. The GUS activity of transgenic plants was
assayed at various developmental stages. The results showed that ZmGLU1
promoter-driven GUS gene had the highest expression level in the roots and that
the expression of GUS gene declined during seed maturation and down to the lowest
level in mature seeds. The ZmGLU1 promoter-driven GUS expression increased during
seed germination, reaching a peak on day 11. The results also showed that this
promoter could be inhibited by 6-BA, trans-zeatin, and NAA, but was not affected
by GA(3), ABA, SA, cold, salt, drought, and submergence treatments. The
histochemical staining revealed that GUS activity was located in vigorous cell
division zones with dominant staining associated with vascular tissues. Deletion
analysis showed that the promoter contained a putative leaf-specific and
stem-specific negative regulative element and two putative enhancers.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16770627 [PubMed - indexed for MEDLINE]

1303: Mol Genet Genomics. 2006 Sep;276(3):242-53. Epub 2006 Jun 10.

Expression of a transcription factor from Capsicum annuum in pine calli
counteracts the inhibitory effects of salt stress on adventitious shoot
formation.

Tang W, Newton RJ, Lin J, Charles TM.

Department of Biology, Howell Science Complex, East Carolina University,
Greenville, NC 27858-4353, USA. tangw@mail.ecu.edu

Transcription factors regulating the stress-responsive gene expression play an
important role in plant stress adaptation. In this study, we examined the salt
stress tolerance of transgenic Virginia pine (Pinus virginiana Mill.)
overexpressing a Capsicum annuum ERF/AP2-type transcription factor (CaPF1), which
may enhance the ability of transgenic plants to tolerate various kinds of
stresses during vegetative growth. CaPF1 transgene increased the salt and
oxidative stress tolerances of pine tissues and counteracted the inhibitory
effects of salt stress on the growth of transgenic Virginia pine calli, shoots,
and plants. To our surprise, the ability of shoot formation was enhanced in three
CaPF1 transgenic Virginia pine cell lines under stress of different NaCl
concentrations. NaCl at 200 mM significantly increased the frequency of
adventitious shoot formation and the number of shoots per gram calli. Measurement
of plant hormone demonstrated that the levels of cytokinin was altered in
CaPF1-overexpressed Virginia pine calli, compared to the control. Based on our
results, we speculate that the altered level of cytokinin may result in enhancing
adventitious shoot formation of transgenic calli exposed to salt for 1 week via
an unknown mechanism.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16767459 [PubMed - indexed for MEDLINE]

1304: Planta. 2006 Oct;224(5):1129-40. Epub 2006 May 16.

Wheat D-type cyclin Triae;CYCD2;1 regulate development of transgenic Arabidopsis
plants.

Wang F, Huo SN, Guo J, Zhang XS.

Shandong Key Laboratory of Crop Biology, College of Life Sciences, Shandong
Agricultural University, Taian, Shandong, 271018, China.

The D-type cyclin genes play key roles in plant development of Arabidopsis. To
investigate the functions of cyclins in monocots, a putative ortholog of cyclin
D2 genes was isolated from wheat shoot tissues, and designated as Triae;CYCD2;1.
The transcripts of Triae;CYCD2;1 were primarily localized in the proliferating
tissues, particularly, in shoot apical meristem and leaf primordia of wheat
plants. Ectopic expression of Triae;CYCD2;1 in Arabidopsis affected plant
morphology and retarded plant growth. Further examination showed that the
promotion of cell division and the inhibition of cell differentiation occurred in
both transgenic plants and tissues. In vitro experiments indicated that
Triae;CYCD2;1 had functional roles in responding to cytokinin and auxin.
Molecular analysis revealed that the transcript levels of several cell
cycle-associated genes, particularly Arath;CYCD3;1, were increased in the
Arabidopsis plants with the expressing Triae;CYCD2;1. The results in this study
provide new information on D-type cyclin in wheat.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16767458 [PubMed - indexed for MEDLINE]

1305: Plant Cell. 2006 Jul;18(7):1652-66. Epub 2006 Jun 9.

Jekyll encodes a novel protein involved in the sexual reproduction of barley.

Radchuk V, Borisjuk L, Radchuk R, Steinbiss HH, Rolletschek H, Broeders S, Wobus
U.

Leibniz-Institut fö¼r Pflanzengenetik und Kulturpflanzenforschung, D-06466
Gatersleben, Germany.

Cereal seed development depends on the intimate interaction of filial and
maternal tissues, ensuring nourishment of the new generation. The gene jekyll,
which was identified in barley (Hordeum vulgare), is preferentially expressed in
the nurse tissues. JEKYLL shares partial similarity with the scorpion Cn4 toxin
and is toxic when ectopically expressed in Escherichia coli and tobacco
(Nicotiana tabacum). In barley, jekyll is upregulated in cells destined for
autolysis. The gene generates a gradient of expression in the nucellar
projection, which mediates the maternal-filial interaction during seed filling.
Downregulation of jekyll by the RNA interference technique in barley decelerates
autolysis and cell differentiation within the nurse tissues. Flower development
and seed filling are thereby extended, and the nucellar projection no longer
functions as the main transport route for assimilates. A slowing down in the
proliferation of endosperm nuclei and a severely impaired ability to accumulate
starch in the endosperm leads to the formation of irregular and small-sized seeds
at maturity. Overall, JEKYLL plays a decisive role in the differentiation of the
nucellar projection and drives the programmed cell death necessary for its proper
function. We further suggest that cell autolysis during the differentiation of
the nucellar projection allows the optimal provision of basic nutrients for
biosynthesis in endosperm and embryo.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16766690 [PubMed - indexed for MEDLINE]

1306: Mol Nutr Food Res. 2006 Jul;50(7):645-54.

Allergen-specific IgE testing in the diagnosis of food allergy and the event of a
positive match in the bioinformatics search.

van Ree R, Vieths S, Poulsen LK.

Academic Medical Center, Amsterdam, The Netherlands.

Current documents on risk assessment of genetically modified foods recommend
including IgE-binding tests on sera from allergic patients. However, there is no
generally accepted recommendation on technical aspects of the testing procedures
or on the interpretation of the results, despite that fact that both false
positive and false-negative results may be caused by variability of the test
procedures. The present article discusses the state-of-the-art of serological
test procedures for qualitative and quantitative determination of specific IgE
and interpretation of test results. It is emphasized that the use of sera from
clinically well-characterized subjects is of high importance. In the case of a
positive test result, the biological activity of the detected IgE antibodies, i.
e., the potential to trigger mediator release from basophils or mast cells in an
allergen-specific manner, should be taken into account. However, present data
also indicate that validation of such mediator release tests is required, both in
terms of experimental protocols and with respect to correlation of the test
results with the clinical situation. Further studies are also required to prove
the usefulness of targeted serum screening, i. e., the testing of gene products
from organisms not known to be allergenic with sera from subjects allergic to
related species.

Publication Types:
Review

PMID: 16764014 [PubMed - indexed for MEDLINE]

1307: Plant Cell Rep. 2006 Nov;25(11):1213-8. Epub 2006 Jun 9.

Development of resistant transgenic soybeans with inverted repeat-coat protein
genes of soybean dwarf virus.

Tougou M, Furutani N, Yamagishi N, Shizukawa Y, Takahata Y, Hidaka S.

National Agricultural Research Center for Tohoku Region, Morioka, Iwate,
020-0198, Japan.

In an attempt to generate soybean plants resistant to soybean dwarf virus (SbDV),
we transformed a construct containing inverted repeat-SbDV coat protein (CP)
genes spaced by beta-glucuronidase (GUS) sequences into soybean somatic embryos
via microprojectile bombardment. Three T(0) plants with an introduced CP gene
were obtained, and one generated T(1) seeds. The presence of the transgene in
T(1) plants was confirmed by PCR and Southern blot hybridization analysis, but
expression of CP was not detected by northern blot hybridization analysis. Two
months after inoculation of SbDV by aphid, T(2) plants contained little
SbDV-specific RNA and remained symptomless. These plants contained
SbDV-CP-specific siRNA. These results suggest that the T(2) plants achieved
resistance to SbDV by an RNA-silencing-mediated process.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16763847 [PubMed - indexed for MEDLINE]

1308: Phytochemistry. 2006 Jun;67(12):1166-76. Epub 2006 Jun 9.

Conjugated fatty acids accumulate to high levels in phospholipids of
metabolically engineered soybean and Arabidopsis seeds.

Cahoon EB, Dietrich CR, Meyer K, Damude HG, Dyer JM, Kinney AJ.

USDA-ARS Plant Genetics Research Unit, Donald Danforth Plant Science Center, 975
North Warson Road, St. Louis, MO 63132, USA. ecahoon@danforthcenter.org

Expression of Delta(12)-oleic acid desaturase-related fatty acid conjugases from
Calendula officinalis, Momordica charantia, and Vernicia fordii in seeds of
soybean (Glycine max) or an Arabidopsis thaliana fad3/fae1 mutant was accompanied
by the accumulation of the conjugated fatty acids calendic acid or
alpha-eleostearic acid to amounts as high as 20% of the total fatty acids.
Conjugated fatty acids, which are synthesized from phosphatidylcholine
(PC)-linked substrates, accumulated in PC and phosphatidylethanolamine, and
relative amounts of these fatty acids were higher in PC than in triacylglycerol
(TAG) in the transgenic seeds. The highest relative amounts of conjugated fatty
acids were detected in PC from seeds of soybean and A. thaliana that expressed
the C. officinalis and M. charantia conjugases, where they accounted for nearly
25% of the fatty acids of this lipid class. In these seeds, >85% of the
conjugated fatty acids in PC were detected in the sn-2 position, and these fatty
acids were also enriched in the sn-2 position of TAG. In marked contrast to the
transgenic seeds, conjugated fatty acids composed <1.5% of the fatty acids in PC
from seeds of five unrelated species that naturally synthesize a variety of
conjugated fatty acid isomers, including seeds that accumulate conjugated fatty
acids to >80% of the total fatty acids. These results suggest that soybean and A.
thaliana seeds are deficient in their metabolic capacity to selectively catalyze
the flux of conjugated fatty acids from their site of synthesis on PC to storage
in TAG.

Publication Types:
Comparative Study

PMID: 16762380 [PubMed - indexed for MEDLINE]

1309: Foodborne Pathog Dis. 2006 Summer;3(2):157-62.

Food safety--who is responsible?

Rollin BE.

Department of Philosophy, Colorado State University, Fort Collins, Colorado
80523-1781, USA. Bernard.Rollin@colostate.edu

Though scientists believe that issues of risk can be handled without appeal to
values in general or ethics in particular, this is demonstrably false. The very
notion of risk is enmeshed in a complex of social ethics. This is clearly true
with regard to food safety. With this in mind, it is plausible to affirm that
responsibility for food safety at a given point in the chain from producer to
consumer rests with the person or entity under whose control the management of
that risk most plausibly lies. This principle is illustrated with various
examples and with clear cases of industry shouldering and avoiding
responsibility. An additional ethical concern relevant to food safety arises from
genetically modified foods. Given that the situation here is uncertain and risk
unknown, it is hard to see who is responsible for managing such risks. It is
arguable that this situation militates in favor of labeling, since consumers are
in effect research subjects. The reasonable moral approach to risk we have
outlined is jeopardized by the societal tendency towards "victimology" and
abrogation of personal responsibility. In such a world, it is incumbent on
industry to educate the public with regard to consumer minimization of food
safety risks, the impossibility of zero-risk situations, and the economic costs
to freedom of protectionism.

PMID: 16761941 [PubMed - indexed for MEDLINE]

1310: Acta Biochim Biophys Sin (Shanghai). 2006 Jun;38(6):393-402.

Rice GTPase OsRacB: potential accessory factor in plant salt-stress signaling.

Luo M, Gu SH, Zhao SH, Zhang F, Wu NH.

Institute of Genetics and Developmental Biology, Chinese Academy of Sciences,
Beijing 100080, China.

As the sole ubiquitous signal small guanosine triphosphate-binding protein in
plants, Rop gene plays an important role in plant growth and development. In this
study, we focus on the relationship between the novel rice Rop gene OsRacB and
plant salt tolerance. Results show that OsRacB transcription is highly
accumulated in roots after treatment with salinity, but only slightly accumulated
in stems and leaves under the same treatment. Promoter analysis showed that
OsRacB promoter is induced by salinity and exogenous salicylic acid, not abscisic
acid. To elucidate its physiological function, we generated OsRacB sense and
antisense transgenic tobacco and rice. Under proper salinity treatment, sense
transgenic plants grew much better than the control. This suggests that
overexpression of OsRacB in tobacco and rice can improve plant salt tolerance.
But under the same treatment, no difference could be observed between OsRacB
antisense plants and the control. The results indicated that OsRacB is only an
accessory factor in plant salt tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16761097 [PubMed - indexed for MEDLINE]

1311: Plant Cell Physiol. 2006 Jul;47(7):915-25. Epub 2006 Jun 7.

Involvement of rice cryptochromes in de-etiolation responses and flowering.

Hirose F, Shinomura T, Tanabata T, Shimada H, Takano M.

Department of Plant Physiology, National Institute of Agrobiological Sciences,
Tsukuba, Ibaraki, 305-8602 Japan.

In order to elucidate the function of cryptochromes (cry) in rice, we have
characterized all rice CRY genes, including OsCRY1a, OsCRY1b and OsCRY2. Our
studies revealed that OsCRY1 genes were mainly expressed in the green plant
tissue, while OsCRY2 gene expression was high in the coleoptile, flower and
callus. Light treatment affected neither the expression of any of the OsCRY genes
nor the stability of their transcripts. However, it was found that Oscry2 protein
was negatively regulated by blue light. Moreover, the level of Oscry2 protein
also decreased upon red light treatment, and this red light-dependent degradation
was shown to be mediated by phytochrome B. Overexpression of OsCRY1 genes
resulted in an increased responsiveness to blue light when measuring coleoptile
growth inhibition. Moreover, growth of leaf sheaths and blades was also repressed
more in OsCRY1 overexpressers than in wild type (WT) under blue light conditions.
These results suggest that Oscry1s are responsible for regulating blue
light-mediated de-etiolation in rice. In addition, OsCRY2 antisense transgenic
rice flowered later than WT under both long-day and short-day conditions,
indicating that Oscry2 is involved in the promotion of flowering time in rice.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16760221 [PubMed - indexed for MEDLINE]

1312: J Theor Biol. 2006 Oct 7;242(3):539-46. Epub 2006 Apr 28.

Invasion of pests resistant to Bt toxins can lead to inherent non-uniqueness in
genetically modified Bt-plant dynamics: mathematical modeling.

Medvinsky AB, Gonik MM, Li BL, Velkov VV, Malchow H.

Institute for Theoretical & Experimental Biophysics, Pushchino, Moscow Region,
142290 Russia. medvinsky@iteb.ru

Genetically modified crops are effective pest management tools for worldwide
growers. However, there is a concern that pests may develop resistance to
Bt-toxins produced by genetically modified Bt-plants. We study the impact of the
Bt-resistant pests on Bt-crops. Furthermore, the dynamics of the
Bt-plant-Bt-susceptible insects-Bt-resistant insects system is analysed and it is
shown that throughout the insect reproduction period the plant biomass dynamics
resulting from invasion of Bt-resistant insects is non-unique. Namely, the
chaotic attractor and the limit cycle, which are responsible for the plant and
insect biomass dynamics, are shown to coexist. As a result, the
Bt-plant-Bt-resistant insect system can manifest either chaotic or regular
oscillations of plant and insect biomass depending on spatial patterns resulting
from invasion of Bt resistant insects into the Bt plant-Bt susceptible insect
system. We show that the non-uniqueness of the system dynamics under unfavorable
environmental conditions, such as in the so-called zones of risky agriculture in
many developing countries and industrialized countries, can lead to essential
decrease in the plant biomass.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16757001 [PubMed - indexed for MEDLINE]

1313: Plant Biol (Stuttg). 2006 Sep;8(5):587-96. Epub 2006 Jun 1.

A novel rice MAPK gene, OsBIMK2, is involved in disease-resistance responses.

Song D, Chen J, Song F, Zheng Z.

Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310029, PR
China.

The mitogen-activated protein kinase (MAPK) cascades play important roles in
transmission of extracellular signals to the downstream effector proteins through
a mechanism of protein phosphorylation. In this study, we isolated and identified
a novel rice MAPK gene, OSBIMK2 ( ORYZAE SATIVA L. BTH-Induced MAP Kinase 2). The
OSBIMK2 encodes a 506 amino acid protein with molecular weight of 63 kD. The
recombinant OSBIMK2 expressed in ESCHERICHIA COLI showed an autophosphorylation
activity IN VITRO. OSBIMK2 is a single-copy gene in the rice genome. Expression
of OSBIMK2 was activated upon treatment with benzothiadiazole (BTH), which is
capable of inducing disease resistance in rice. Expression of OsBIMK2 was also
up-regulated during early stage after inoculation with MAGNAPORTHE GRISEA in
BTH-treated rice seedlings and during an incompatible interaction between M.
GRISEA and a blast-resistant rice genotype. Over-expression of the rice OSBIMK2
gene in transgenic tobacco resulted in an enhanced disease resistance against
tomato mosaic virus and a fungal pathogen, ALTERNARIA ALTERNATA. These results
suggest that OSBIMK2 plays a role in disease resistance responses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16755461 [PubMed - indexed for MEDLINE]

1314: Planta. 2006 Oct;224(5):1116-28. Epub 2006 Jun 2.

Molecular and phenotypic characterization of transgenic soybean expressing the
Arabidopsis ferric chelate reductase gene, FRO2.

Vasconcelos M, Eckert H, Arahana V, Graef G, Grusak MA, Clemente T.

USDA/ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor
College of Medicine, Houston, TX, USA.

Soybean (Glycine max Merr.) production is reduced under iron-limiting calcareous
soils throughout the upper Midwest regions of the US. Like other dicotyledonous
plants, soybean responds to iron-limiting environments by induction of an active
proton pump, a ferric iron reductase and an iron transporter. Here we demonstrate
that heterologous expression of the Arabidopsis thaliana ferric chelate reductase
gene, FRO2, in transgenic soybean significantly enhances Fe(+3) reduction in
roots and leaves. Root ferric reductase activity was up to tenfold higher in
transgenic plants and was not subjected to post-transcriptional regulation. In
leaves, reductase activity was threefold higher in the transgenic plants when
compared to control. The enhanced ferric reductase activity led to reduced
chlorosis, increased chlorophyll concentration and a lessening in biomass loss in
the transgenic events between Fe treatments as compared to control plants grown
under hydroponics that mimicked Fe-sufficient and Fe-deficient soil environments.
However, the data indicate that constitutive FRO2 expression under non-iron
stress conditions may lead to a decrease in plant productivity as reflected by
reduced biomass accumulation in the transgenic events under non-iron stress
conditions. When grown at Fe(III)-EDDHA levels greater than 10 microM, iron
concentration in the shoots of transgenic plants was significantly higher than
control. The same observation was found in the roots in plants grown at iron
levels higher than 32 microM Fe(III)-EDDHA. These results suggest that
heterologous expression of an iron chelate reductase in soybean can provide a
route to alleviate iron deficiency chlorosis.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16741749 [PubMed - indexed for MEDLINE]

1315: J Environ Qual. 2006 May 31;35(4):1001-9. Print 2006 Jul-Aug.

Impact of transgenic Bt maize residues on the mycotoxigenic plant pathogen
Fusarium graminearum and the biocontrol agent Trichoderma atroviride.

Naef A, Zesiger T, Dö©fago G.

Plant Pathology, Institute of Integrative Biology, ETH Zurich, 8092 Zurich,
Switzerland.

Transformation of maize with genes encoding for insecticidal crystal (Cry)
proteins from Bacillus thuringiensis (Bt) could have an impact on the saprophytic
survival of plant pathogens and their antagonists on crop residues. We assessed
potential effects on the mycotoxin deoxynivalenol (DON)-producing wheat and maize
pathogen Fusarium graminearum and on the biocontrol agent Trichoderma atroviride.
Purified Cry1Ab protein caused no growth inhibition of these fungi on agar
plates. Cry1Ab concentrations above levels common in Bt maize tissue stimulated
the growth of F. graminearum. The fungi were also grown on
gamma-radiation-sterilized leaf tissue of four Bt maize hybrids and their non
transgenic isolines collected at maize maturity on a field trial in 2002 and
2003. Both fungi degraded the Cry1Ab protein in Bt maize tissue. Fungal biomass
quantification with microsatellite-based polymerase chain reaction (PCR) assays
revealed differential fungal growth on leaf tissue of different maize varieties
but no consistent difference between corresponding Bt and non-Bt hybrids.
Generally, year of maize tissue collection had a greater impact on biomass
production than cultivar or Bt transformation. The mycotoxin DON levels observed
in maize tissue experiments corresponded with patterns in F. graminearum biomass,
indicating that Bt transformation has no impact on DON production. In addition to
bioassays, maize leaf tissue was analyzed with a mass spectrometer-based
electronic nose, generating fingerprints of volatile organic compounds. Chemical
fingerprints of corresponding Bt and non-Bt leaf tissues differed only for those
hybrid pairs that caused differential fungal biomass production in the bioassays.
Our results suggest that Cry1Ab protein in maize residues has no direct effect on
F. graminearum and T. atroviride but some corresponding Bt/non-Bt maize hybrids
differ more in composition than Cry protein content alone, which can affect the
saprophytic growth of fungi on crop residues.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16738384 [PubMed - indexed for MEDLINE]

1316: Mol Nutr Food Res. 2006 Jul;50(7):604-9.

Review of the development of methodology for evaluating the human allergenic
potential of novel proteins.

Taylor SL.

University of Nebraska, Food Allergy Research & Resource Program, Lincoln,
Nebraska 68583, USA. staylor2@unl.edu

Safety assessment of novel proteins in genetic-engineered foods is a key
component of the overall safety evaluation for these products. Since allergens
are typically proteins, assessment of the potential allergenicity of the novel
proteins in genetically engineered foods is critical. This article reviews
methods available to assess the potential allergenicity of novel proteins, as
well as problems and deficiencies in the existing methods. The role of
bioinformatics and knowledge of allergenic epitopes in developing new approaches
to this problem is discussed.

Publication Types:
Comparative Study
Review

PMID: 16736463 [PubMed - indexed for MEDLINE]

1317: Plant Cell. 2006 Jul;18(7):1642-51. Epub 2006 May 26.

A network of local and redundant gene regulation governs Arabidopsis seed
maturation.

To A, Valon C, Savino G, Guilleminot J, Devic M, Giraudat J, Parcy F.

Institut des Sciences du Vö©gö©tal, Unitö© Propre de Recherche 2355, Centre National
de la Recherche Scientifique, 91190 Gif-sur-Yvette, France.

In Arabidopsis thaliana, four major regulators (ABSCISIC ACID INSENSITIVE3
[ABI3], FUSCA3 [FUS3], LEAFY COTYLEDON1 [LEC1], and LEC2) control most aspects of
seed maturation, such as accumulation of storage compounds, cotyledon identity,
acquisition of desiccation tolerance, and dormancy. The molecular basis for
complex genetic interactions among these regulators is poorly understood. By
analyzing ABI3 and FUS3 expression in various single, double, and triple
maturation mutants, we have identified multiple regulatory links among all four
genes. We found that one of the major roles of LEC2 was to upregulate FUS3 and
ABI3. The lec2 mutation is responsible for a dramatic decrease in ABI3 and FUS3
expression, and most lec2 phenotypes can be rescued by ABI3 or FUS3 constitutive
expression. In addition, ABI3 and FUS3 positively regulate themselves and each
other, thereby forming feedback loops essential for their sustained and uniform
expression in the embryo. Finally, LEC1 also positively regulates ABI3 and FUS3
in the cotyledons. Most of the genetic controls discovered were found to be local
and redundant, explaining why they had previously been overlooked. This works
establishes a genetic framework for seed maturation, organizing the key
regulators of this process into a hierarchical network. In addition, it offers a
molecular explanation for the puzzling variable features of lec2 mutant embryos.

PMID: 16731585 [PubMed - indexed for MEDLINE]

1318: PLoS Biol. 2006 Jun;4(6):e181.

Resistance evolution to Bt crops: predispersal mating of European corn borers.

Dalecky A, Ponsard S, Bailey RI, Pö©lissier C, Bourguet D.

Institut National de la Recherche Agronomique, UMR Centre de Biologie et de
Gestion des Populations, Campus International de Baillarguet,
Montferrier-sur-Lez, France.

Over the past decade, the high-dose refuge (HDR) strategy, aimed at delaying the
evolution of pest resistance to Bacillus thuringiensis (Bt) toxins produced by
transgenic crops, became mandatory in the United States and is being discussed
for Europe. However, precopulatory dispersal and the mating rate between resident
and immigrant individuals, two features influencing the efficiency of this
strategy, have seldom been quantified in pests targeted by these toxins. We
combined mark-recapture and biogeochemical marking over three breeding seasons to
quantify these features directly in natural populations of Ostrinia nubilalis, a
major lepidopteran corn pest. At the local scale, resident females mated
regardless of males having dispersed beforehand or not, as assumed in the HDR
strategy. Accordingly, 0-67% of resident females mating before dispersal did so
with resident males, this percentage depending on the local proportion of
resident males (0% to 67.2%). However, resident males rarely mated with immigrant
females (which mostly arrived mated), the fraction of females mating before
dispersal was variable and sometimes substantial (4.8% to 56.8%), and there was
no evidence for male premating dispersal being higher. Hence, O. nubilalis
probably mates at a more restricted spatial scale than previously assumed, a
feature that may decrease the efficiency of the HDR strategy under certain
circumstances, depending for example on crop rotation practices.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16719560 [PubMed - indexed for MEDLINE]

1319: Planta. 2006 Oct;224(5):1050-7. Epub 2006 May 23.

Co-expression of the borage Delta 6 desaturase and the Arabidopsis Delta 15
desaturase results in high accumulation of stearidonic acid in the seeds of
transgenic soybean.

Eckert H, La Vallee B, Schweiger BJ, Kinney AJ, Cahoon EB, Clemente T.

Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln,
NE 68588-0665, USA.

Two relatively rare fatty acids, gamma-linolenic acid (GLA) and stearidonic acid
(STA), have attracted much interest due to their nutraceutical and pharmaceutical
potential. STA, in particular, has been considered a valuable alternative source
for omega-3 fatty acids due to its enhanced conversion efficiency in animals to
eicosapentaenoic acid when compared with the more widely consumed omega-3 fatty
acid, alpha-linolenic acid (ALA), present in most vegetable oils. Exploiting the
wealth of information currently available on in planta oil biosynthesis and
coupling this information with the tool of genetic engineering it is now feasible
to deliberately perturb fatty acid pools to generate unique oils in commodity
crops. In an attempt to maximize the STA content of soybean oil, a borage
Delta(6) desaturase and an Arabidopsis Delta(15) desaturase were pyramided by
either sexual crossing of transgenic events, re-transformation of a Delta(6)
desaturase event with the Delta(15) desaturase or co-transformation of both
desaturases. Expression of both desaturases in this study was under the control
of the seed-specific soybean beta-conglycinin promoter. Soybean events that
carried only the Delta(15 )desaturase possessed a significant elevation of ALA
content, while events with both desaturases displayed a relative STA abundance
greater than 29%, creating a soybean with omega-3 fatty acids representing over
60% of the fatty acid profile. Analyses of the membrane lipids in a subset of the
transgenic events suggest that soybean seeds compensate for enhanced production
of polyunsaturated fatty acids by increasing the relative content of palmitic
acid in phosphatidylcholine and other phospholipids.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16718484 [PubMed - indexed for MEDLINE]

1320: Planta. 2006 Oct;224(5):1209-25. Epub 2006 May 23.

Functional roles of the pepper pathogen-induced bZIP transcription factor,
CAbZIP1, in enhanced resistance to pathogen infection and environmental stresses.

Lee SC, Choi HW, Hwang IS, Choi du S, Hwang BK.

Laboratory of Molecular Plant Pathology, College of Life Sciences and
Biotechnology, Korea University, Seoul, 136-713, South Korea.

Transcription factors often belong to multigene families and their individual
contribution in a particular regulatory network remains difficult to assess. We
identify and functionally characterize the pepper bZIP transcription factor
CAbZIP1 gene isolated from pepper leaves infected with Xanthomonas campestris pv.
vesicatoria. Transient expression analysis of the CAbZIP1-GFP fusion protein in
Arabidopsis protoplasts revealed that the CAbZIP1 protein is localized in the
nucleus. The N-terminal region of CAbZIP1 fused to the GAL4 DNA-binding domain is
required to activate transcription of reporter genes in yeast. The CAbZIP1
transcripts are constitutively expressed in the pepper root and flower, but not
in the leaf, stem and fruit. The CAbZIP1 gene is locally or systemically induced
in pepper plants infected by either X. campestris pv. vesicatoria or Pseudomonas
fluorescens. The CAbZIP1 gene is also induced by abiotic elicitors and
environmental stresses. The CAbZIP1 transgenic Arabidopsis exhibits a dwarf
phenotype, indicating that CAbZIP1 may be involved in plant development. The
CAbZIP1 overexpression in the transgenic Arabidopsis plants confers enhanced
resistance to Pseudomonas syringae pv. tomato DC3000, accompanied by expression
of the AtPR-4 and AtRD29A. The transgenic plants also exhibit increased drought
and salt tolerance during all growth stages. Moreover, the transgenic plants are
tolerant to methyl viologen-oxidative stress. Together, these data suggest that
the CAbZIP1 transcription factor function as a possible regulator in enhanced
disease resistance and environmental stress tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16718483 [PubMed - indexed for MEDLINE]

1321: J Biotechnol. 2006 Jul 13;124(2):469-72. Epub 2006 May 23.

High level expression of bioactive recombinant human growth hormone in the milk
of a cloned transgenic cow.

Salamone D, Baraö±ao L, Santos C, Bussmann L, Artuso J, Werning C, Prync A,
Carbonetto C, Dabsys S, Munar C, Salaberry R, Berra G, Berra I, Fernö¡ndez N,
Papouchado M, Foti M, Judewicz N, Mujica I, Muö±oz L, Alvarez SF, Gonzö¡lez E,
Zimmermann J, Criscuolo M, Melo C.

Biosidus SA, Buenos Aires, Argentina.

Transgenic farm animals have been proposed as an alternative to current
bioreactors for large scale production of biopharmaceuticals. However, the
efficiency of both methods in the production of the same protein has not yet been
established. Here we report the production of recombinant human growth hormone
(hGH) in the milk of a cloned transgenic cow at levels of up to 5 g l(-1). The
hormone is identical to that currently produced by expression in E. coli. In
addition, the hematological and somatometric parameters of the cloned transgenic
cow are within the normal range for the breed and it is fertile and capable of
producing normal offspring. These results demonstrate that transgenic cattle can
be used as a cost-effective alternative for the production of this hormone.

PMID: 16716426 [PubMed - indexed for MEDLINE]

1322: Plant Physiol. 2006 Jul;141(3):924-31. Epub 2006 May 19.

Morphological alteration caused by brassinosteroid insensitivity increases the
biomass and grain production of rice.

Morinaka Y, Sakamoto T, Inukai Y, Agetsuma M, Kitano H, Ashikari M, Matsuoka M.

Bioscience and Biotechnology Center, Nagoya University, Nagoya, Aichi 464-8601,
Japan.

The rice (Oryza sativa) dwarf mutant d61 phenotype is caused by loss of function
of a rice BRASSINOSTEROID INSENSITIVE1 ortholog, OsBRI1. We have identified nine
d61 alleles, the weakest of which, d61-7, confers agronomically important traits
such as semidwarf stature and erect leaves. Because erect-leaf habit is
considered to increase light capture for photosynthesis, we compared the biomass
and grain production of wild-type and d61-7 rice. The biomass of wild type was
38% higher than that of d61-7 at harvest under conventional planting density
because of the dwarfism of d61-7. However, the biomass of d61-7 was 35% higher
than that of wild type at high planting density. The grain yield of wild type
reached a maximum at middensity, but the yield of d61-7 continued to increase
with planting density. These results indicate that d61-7 produces biomass more
effectively than wild type, and consequently more effectively assimilates the
biomass in reproductive organ development at high planting density. However, the
small grain size of d61-7 counters any increase in grain yield, leading to the
same grain yield as that of wild type even at high density. We therefore produced
transgenic rice with partial suppression of endogenous OsBRI1 expression to
obtain the erect-leaf phenotype without grain changes. The estimated grain yield
of these transformants was about 30% higher than that of wild type at high
density. These results demonstrate the feasibility of generating erect-leaf
plants by modifying the expression of the brassinosteroid receptor gene in
transgenic rice plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16714407 [PubMed - indexed for MEDLINE]

1323: Plant Cell Rep. 2006 Oct;25(10):1024-34. Epub 2006 May 19.

Improved Agrobacterium-mediated transformation of three maize inbred lines using
MS salts.

Frame BR, McMurray JM, Fonger TM, Main ML, Taylor KW, Torney FJ, Paz MM, Wang K.

Department of Agronomy, Iowa State University, Ames, IA 50011-1010, USA.

Transformation technology as a research or breeding tool to improve maize is
routinely used in most industrial and some specialized public laboratories.
However, transformation of many inbred lines remains a challenging task,
especially when using Agrobacterium tumefaciens as the delivery method. Here we
report success in generating transgenic plants and progeny from three maize
inbred lines using an Agrobacterium-mediated standard binary vector system to
target maize immature embryos. Eleven maize inbred lines were pre-screened for
transformation frequency using N6 salts. A subset of three maize inbred lines was
then systematically evaluated for frequency of post-infection embryogenic callus
induction and transformation on four media regimes: N6 or MS salts in each of two
distinct media backgrounds. Transgenic plants recovered from inbred lines B104,
B114, and Ky21 were analyzed for transgene integration, expression, and
transmission. Average transformation frequencies of 6.4% (for B104), 2.8% (for
B114), and 8% (for Ky21) were achieved using MS salts. Availability of
Agrobacterium-mediated maize inbred line transformation will improve future
opportunities for maize genetic and functional genomic studies.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16710703 [PubMed - indexed for MEDLINE]

1324: Plant J. 2006 Jun;46(5):880-9.

Identification and characterization of Arabidopsis gibberellin receptors.

Nakajima M, Shimada A, Takashi Y, Kim YC, Park SH, Ueguchi-Tanaka M, Suzuki H,
Katoh E, Iuchi S, Kobayashi M, Maeda T, Matsuoka M, Yamaguchi I.

Department of Applied Biological Chemistry, The University of Tokyo, Tokyo
113-8657, Japan. nkjm@pgr1.ch.a.u-tokyo.ac.jp

Three gibberellin (GA) receptor genes (AtGID1a, AtGID1b and AtGID1c), each an
ortholog of the rice GA receptor gene (OsGID1), were cloned from Arabidopsis, and
the characteristics of their recombinant proteins were examined. The GA-binding
activities of the three recombinant proteins were confirmed by an in vitro assay.
Biochemical analyses revealed similar ligand selectivity among the recombinants,
and all recombinants showed higher affinity to GA(4) than to other GAs. AtGID1b
was unique in its binding affinity to GA(4) and in its pH dependence when
compared with the other two, by only showing binding in a narrow pH range (pH
6.4-7.5) with 10-fold higher affinity (apparent K(d) for GA(4) = 3 x 10(-8) m)
than AtGID1a and AtGID1c. A two-hybrid yeast system only showed in vivo
interaction in the presence of GA(4) between each AtGID1 and the Arabidopsis
DELLA proteins (AtDELLAs), negative regulators of GA signaling. For this
interaction with AtDELLAs, AtGID1b required only one-tenth of the amount of GA(4)
that was necessary for interaction between the other AtGID1s and AtDELLAs,
reflecting its lower K(d) value. AtDELLA boosted the GA-binding activity of
AtGID1 in vitro, which suggests the formation of a complex between AtDELLA and
AtGID1-GA that binds AtGID1 to GA more tightly. The expression of each AtGID1
clone in the rice gid1-1 mutant rescued the GA-insensitive dwarf phenotype. These
results demonstrate that all three AtGID1s functioned as GA receptors in
Arabidopsis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16709201 [PubMed - indexed for MEDLINE]

1325: Plant J. 2006 Jun;46(5):834-48.

Over-expression of SOB5 suggests the involvement of a novel plant protein in
cytokinin-mediated development.

Zhang J, Wrage EL, Vankova R, Malbeck J, Neff MM.

Department of Biology, Washington University, Campus Box 1137, One Brookings
Drive, St Louis, MO 63130, USA.

Cytokinins are a class of phytohormones that play a critical role in plant growth
and development. sob5-D, an activation-tagging mutant, shows phenotypes typical
of transgenic plants expressing the Agrobacterium tumefaciens
isopentenyltransferase (ipt) gene that encodes the enzyme catalyzing the first
step of cytokinin biosynthesis. The sob5-D mutant phenotypes are caused by
over-expression of a novel gene, SOB5. Sequence analysis places SOB5 in a
previously uncharacterized family of plant-specific proteins. A translational
fusion between SOB5 and the green fluorescent protein reporter was localized in
the cytoplasm as well as associated with the plasma membrane when transiently
expressed in onion epidermal cells. Analysis of transgenic plants harboring an
SOB5:SOB5-beta-glucuronidase (GUS) translational fusion under the control of the
SOB5 promoter region showed GUS activity in vegetative tissues (hydathodes and
trichomes of leaves, shoot meristems and roots) as well as in floral tissues
(pistil tips, developing anthers and sepal vasculature). Cytokinin quantification
analysis revealed that adult sob5-D plants accumulated higher levels of
trans-zeatin riboside, trans-zeatin riboside monophosphate and isopentenyladenine
9-glucoside when compared to the wild-type. Consistent with this result, AtIPT3
and AtIPT7 were found to be up-regulated in a tissue-specific manner in sob5-D
mutants. Physiological analysis of the sob5-D mutant demonstrated reduced
responsiveness to exogenous cytokinin in both root-elongation and
callus-formation assays. Taken together, our data suggest a role for the novel
gene SOB5 in cytokinin-mediated plant development.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16709198 [PubMed - indexed for MEDLINE]

1326: Plant J. 2006 Jun;46(5):747-57.

Maize DBF1-interactor protein 1 containing an R3H domain is a potential regulator
of DBF1 activity in stress responses.

Saleh A, Lumbreras V, Lopez C, Dominguez-Puigjaner E, Kizis D, Pagö¨s M.

Departamento de Genö©tica Molecular, Instituto de Biologö­a Molecular de Barcelona,
IBMB, Consejo Superior de Investigaciones Cientö­ficas, CSIC, 18-26 Jordi Girona,
08034 Barcelona, Spain.

The maize dehydration-responsive element (DRE)-binding factor, DBF1, is a member
of the Apetala 2/Ethylene Response Factor transcription factors family and is
involved in the regulation of the ABA-responsive gene rab17 through the DRE in an
ABA-dependent pathway. In this study we analysed the functionality of DBF1 in
abiotic stress responses and found that Arabidopsis plants over-expressing DBF1
were more tolerant to osmotic stress than control plants. In yeast two-hybrid
analyses, DBF1 interacted with DBF1-interactor protein 1 (DIP1), a protein
containing a conserved R3H single-strand DNA-binding domain. Subcellular
localization of DIP1 showed that the protein fusion DIP1-Red Flourescent Protein
(RFP) was mainly localized in the cytoplasm. However, after co-transformation of
DBF1-GFP and DIP1-RFP, both proteins co-localized in the nucleus. Interestingly,
when the N-terminal DBF1-GFP was co-expressed with DIP1-RFP, both proteins
co-localized predominantly in the cytoplasmic speckles observed for N-terminal
DBF1-GFP fusion protein. These results clearly show in vivo interaction of DBF1
with DIP1 in the cell and that this interaction is necessary for the nuclear
localization of DIP1 protein. Analysis of the regulatory effect of the DBF1 and
DIP1 interaction on the maize rab17 promoter activity indicated that
co-transfection of DBF1 with DIP1 enhances promoter activity in the absence of
ABA treatment. We suggest that the regulated association of DBF1 and DIP1 may
control the levels of target gene expression during stress conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16709191 [PubMed - indexed for MEDLINE]

1327: Plant J. 2006 Jun;46(5):723-35.

Gene duplication, exon gain and neofunctionalization of OEP16-related genes in
land plants.

Drea SC, Lao NT, Wolfe KH, Kavanagh TA.

Plant Molecular Genetics Laboratory, Smurfit Institute of Genetics, Trinity
College, Dublin 2, Ireland.

OEP16, a channel protein of the outer membrane of chloroplasts, has been
implicated in amino acid transport and in the substrate-dependent import of
protochlorophyllide oxidoreductase A. Two major clades of OEP16-related sequences
were identified in land plants (OEP16-L and OEP16-S), which arose by a gene
duplication event predating the divergence of seed plants and bryophytes.
Remarkably, in angiosperms, OEP16-S genes evolved by gaining an additional exon
that extends an interhelical loop domain in the pore-forming region of the
protein. We analysed the sequence, structure and expression of the corresponding
Arabidopsis genes (atOEP16-S and atOEP16-L) and demonstrated that following
duplication, both genes diverged in terms of expression patterns and coding
sequence. AtOEP16-S, which contains multiple G-box ABA-responsive elements
(ABREs) in the promoter region, is regulated by ABI3 and ABI5 and is strongly
expressed during the maturation phase in seeds and pollen grains, both
desiccation-tolerant tissues. In contrast, atOEP-L, which lacks promoter ABREs,
is expressed predominantly in leaves, is induced strongly by low-temperature
stress and shows weak induction in response to osmotic stress, salicylic acid and
exogenous ABA. Our results indicate that gene duplication, exon gain and
regulatory sequence evolution each played a role in the divergence of OEP16
homologues in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16709189 [PubMed - indexed for MEDLINE]

1328: Proc Natl Acad Sci U S A. 2006 May 23;103(21):8287-92. Epub 2006 May 12.

Tomato aromatic amino acid decarboxylases participate in synthesis of the flavor
volatiles 2-phenylethanol and 2-phenylacetaldehyde.

Tieman D, Taylor M, Schauer N, Fernie AR, Hanson AD, Klee HJ.

Department of Horticultural Sciences, University of Florida, P.O. Box 110690,
Gainesville, FL 32611-0690, USA.

An important phenylalanine-derived volatile compound produced by plants is
2-phenylethanol. It is a major contributor to flavor in many foods, including
fresh fruits, such as tomato, and an insect-attracting scent in roses and many
other flowers. Despite the centrality of 2-phenylethanol to flavor and fragrance,
the plant genes responsible for its synthesis have not been identified. Here, we
describe a biosynthetic pathway for 2-phenylethanol and other
phenylalanine-derived volatiles in tomato fruits and a small family of
decarboxylases (LeAADC1A, LeAADC1B, and LeAADC2) that can mediate that pathway's
first step. These enzymes each catalyze conversion of phenylalanine to
phenethylamine and tyrosine to tyramine. Although tyrosine is the preferred
substrate in vitro, phenylalanine levels in tomato fruits far exceed those of
tyrosine, indicating that phenylalanine is a physiological substrate. Consistent
with this view, overexpression of either LeAADC1A or LeAADC2 in transgenic tomato
plants resulted in fruits with up to 10-fold increased emissions of the products
of the pathway, including 2-phenylacetaldehyde, 2-phenylethanol, and
1-nitro-2-phenylethane. Further, antisense reduction of LeAADC2 significantly
reduced emissions of these volatiles. Besides establishing a biosynthetic route,
these results show that it is possible to change phenylalanine-based flavor and
aroma volatiles in plants by manipulating expression of a single gene.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16698923 [PubMed - indexed for MEDLINE]

1329: Plant Physiol. 2006 Jul;141(3):898-909. Epub 2006 May 12.

Erratum in:
Plant Physiol. 2006 Dec;142(4):1771.

Surface position, not signaling from surrounding maternal tissues, specifies
aleurone epidermal cell fate in maize.

Gruis DF, Guo H, Selinger D, Tian Q, Olsen OA.

Pioneer Hi-Bred International, a DuPont Company, Johnston, Iowa 50131, USA.
fred.gruis@pioneer.com

Maize (Zea mays) endosperm consists of an epidermal-like surface layer of
aleurone cells, an underlying body of starchy endosperm cells, and a basal layer
of transfer cells. To determine whether surrounding maternal tissues perform a
role in specifying endosperm cell fates, a maize endosperm organ culture
technique was established whereby the developing endosperm is completely removed
from surrounding maternal tissues. Using cell type-specific fluorescence markers,
we show that aleurone cell fate specification occurs exclusively in response to
surface position and does not require specific, continued maternal signal input.
The starchy endosperm and aleurone cell fates are freely interchangeable
throughout the lifespan of the endosperm, with internalized aleurone cells
converting to starchy endosperm cells and with starchy endosperm cells that
become positioned at the surface converting to aleurone cells. In contrast to
aleurone and starchy endosperm cells, transfer cells fail to develop in in
vitro-grown endosperm, supporting earlier indications that maternal tissue
interaction is required to fully differentiate this cell type. Several parameters
confirm that the maize endosperm organ cultures described herein retain the main
developmental features of in planta endosperm, including fidelity of aleurone
mutant phenotypes, temporal and spatial control of cell type-specific fluorescent
markers, specificity of cell type transcripts, and control of mitotic cell
divisions.

PMID: 16698897 [PubMed - indexed for MEDLINE]

1330: FEBS Lett. 2006 May 29;580(13):3315-20. Epub 2006 May 8.

A transgenic rice seed accumulating an anti-hypertensive peptide reduces the
blood pressure of spontaneously hypertensive rats.

Yang L, Tada Y, Yamamoto MP, Zhao H, Yoshikawa M, Takaiwa F.

Transgenic Crop Research and Development Center, National Institute of
Agrobiological Sciences (NIAS), Tsukuba, Ibaraki 305-8602, Japan.

RPLKPW is a potent anti-hypertensive peptide designed according to the structure
of ovokinin(2-7) (RADHPF). In this study, we generated transgenic rice plants
that accumulate the RPLKPW peptide as a fusion protein with the rice storage
protein glutelin. The engineered peptide is expressed under the control of
endosperm-specific glutelin promoters and specifically accumulates in seeds. Oral
administration of either the RPLKPW-glutelin fraction or transgenic rice seeds to
spontaneously hypertensive rats (SHRs) significantly reduced systolic blood
pressures. These results suggest the possible application of transgenic rice seed
as a nutraceutical delivery system and specifically for administration of active
peptides in hypertension.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16697378 [PubMed - indexed for MEDLINE]

1331: Trends Plant Sci. 2006 Jun;11(6):261-3. Epub 2006 May 11.

Wild sex in the grasses.

Able JA, Langridge P.

Molecular Plant Breeding Cooperative Research Centre, School of Agriculture, Food
& Wine, The University of Adelaide, Glen Osmond, SA 5064, Australia.
jason.able@adelaide.edu.au

To date, alien introgression of agronomically important traits into bread wheat
(Triticum aestivum) from wild relatives has not been readily achievable through
traditional breeding practices. However, this door might now be unlocked. The
insightful research published recently by Graham Moore and his team delivers a
likely candidate in the form of a cdc2-kinase-related gene family for the Ph1
locus--a chromatin region located on chromosome 5B that is responsible for
homologous chromosome pairing integrity in bread wheat.

PMID: 16697246 [PubMed - indexed for MEDLINE]

1332: Physiol Behav. 2006 Nov 30;89(4):465-71. Epub 2006 May 11.

Amylinergic control of food intake.

Lutz TA.

Institute of Veterinary Physiology, Vetsuisse Faculty University of Zurich and
Center of Integrative Human Physiology, Winterthurerstrasse 260, 8057 Zurich,
Switzerland. tomlutz@vetphys.unizh.ch

Amylin is a pancreatic B-cell hormone that plays an important role in the
regulation of nutrient fluxes. As such, amylin reduces food intake in laboratory
animals and man, slows gastric emptying and it reduces postprandial glucagon
secretion. Amylin deficiency which occurs concomitantly to insulin deficiency in
diabetes mellitus, may therefore contribute to some of the major derangements
associated with this disorder (hyperphagia, excessive glucagon secretion,
accelerated rate of gastric emptying). The described actions of amylin all seem
to depend on a direct effect of amylin on the area postrema (AP). As to amylin's
satiating effect, the physiological relevance of this action is underlined by
studies involving specific amylin antagonists and amylin-deficient mice. In the
AP, amylin seems to modulate the anorectic signal elicited by CCK. Subsequent to
AP activation, the amylin signal is conveyed to the forebrain via distinct relay
stations. Within the lateral hypothalamic area, amylin diminishes the expression
of orexigenic neuropeptides such as orexin and MCH. Whether these effects
contribute to amylin's short term satiating action remains to be determined.
Recent studies suggest that amylin may also play a role as a long-term,
lipostatic signal, especially when other feedback systems to the brain are
deficient. Obese, leptin-resistant Zucker rats which are hyperinsulinemic and
hyperamylinemic, were chronically infused with the amylin antagonist AC 187. AC
187 significantly elevated food intake in obese Zucker rats while having no
effect in lean controls. This indicates that at least under certain conditions,
chronic blockade of endogenous amylin action may lead to an increase in food
intake and/or body weight. As mentioned, the site and mechanism of action for
peripheral amylin to reduce food intake seems to be well established. It is less
clear how centrally administered amylin reduces food intake although it is well
known that 3rd ventricular administration of amylin produces a very strong and
long-lasting anorectic action. Amylin receptors have been described in various
hypothalamic nuclei but the endogenous ligand of these receptors remains to be
investigated. The same holds true as to the physiological relevance of the
anorectic effect seen after central amylin administration.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 16697020 [PubMed - indexed for MEDLINE]

1333: FEBS Lett. 2006 May 29;580(13):3136-44. Epub 2006 May 2.

Constitutive expression of abiotic stress-inducible hot pepper CaXTH3, which
encodes a xyloglucan endotransglucosylase/hydrolase homolog, improves drought and
salt tolerance in transgenic Arabidopsis plants.

Cho SK, Kim JE, Park JA, Eom TJ, Kim WT.

Department of Biology, College of Science, Yonsei University, Seoul 120-749,
Republic of Korea.

Xyloglucan endotransglucosylase/hydrolase (XTH) has been recognized as a cell
wall-modifying enzyme, participating in the diverse physiological roles. From
water-stressed hot pepper plants, we isolated three different cDNA clones
(pCaXTH1, pCaXTH2, and pCaXTH3) that encode XTH homologs. RT-PCR analysis showed
that three CaXTH mRNAs were concomitantly induced by a broad spectrum of abiotic
stresses, including drought, high salinity and cold temperature, and in response
to stress hormone ethylene, suggesting their role in the early events in the
abiotic-related defense response. Transgenic Arabidopsis plants that
constitutively expressed the CaXTH3 gene under the control of the CaMV 35S
promoter exhibited abnormal leaf morphology; the transgenic leaves showed
variable degrees of twisting and bending along the edges, resulting in a severely
wrinkled leaf shape. Microscopic analysis showed that 35S-CaXTH3 leaves had
increased numbers of small-sized cells, resulting in disordered, highly populated
mesophyll cells in each dorsoventral layer, and appeared to contain a limited
amount of starch. In addition, the 35S-CaXTH3 transgenic plants displayed
markedly improved tolerance to severe water deficit, and to lesser extent to high
salinity in comparison with the wild-type plants. These results indicate that
CaXTH3 is functional in heterologous Arabidopsis cells, thereby effectively
altering cell growth and also the response to abiotic stresses. Although the
physiological function of CaXTHs is not yet clear, there are several
possibilities for their involvement in a subset of physiological responses to
counteract dehydration and high salinity stresses in transgenic Arabidopsis
plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16684525 [PubMed - indexed for MEDLINE]

1334: Trends Biotechnol. 2006 Jul;24(7):305-11. Epub 2006 May 6.

Genetic engineering of wheat--current challenges and opportunities.

Bhalla PL.

Plant Molecular Biology and Biotechnology Laboratory, Australian Research Council
Centre of Excellence for Integrative Legume Research, The University of
Melbourne, Parkville, Victoria 3010, Australia. premlb@unimelb.edu.au

Wheat is one of the major staple food crops grown worldwide; however,
productivity in cereal crops has not kept pace with the world population growth.
A significant increase in wheat production (>40% by 2020) is needed simply to
keep up with the growing demand. This increase is unlikely to be achieved by
conventional plant breeding methods because of the limited gene pool available.
The application of recombinant techniques to improve wheat quality and yield is
not only desirable but also has potential to open up new opportunities. Although
there has been significant progress in developing gene-transformation
technologies for improving these traits, this remains an important challenge for
plant biotechnology. Obstacles to translate the full potential of the genomic era
to wheat breeding include the need to develop elite wheat varieties without
selectable markers, introducing minimal or nil intergenic DNA and social and
market issues concerning genetically engineered food products.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 16682090 [PubMed - indexed for MEDLINE]

1335: Plant Physiol. 2006 Jul;141(3):910-23. Epub 2006 May 5.

Proteomic investigation of the effect of salicylic acid on Arabidopsis seed
germination and establishment of early defense mechanisms.

Rajjou L, Belghazi M, Huguet R, Robin C, Moreau A, Job C, Job D.

Centre National de la Recherche Scientifique, Bayer CropScience Joint Laboratory,
Unitö© Mixte de Recherche 2847, F-69263 Lyon cedex 09, France.

The influence of salicylic acid (SA) on elicitation of defense mechanisms in
Arabidopsis (Arabidopsis thaliana) seeds and seedlings was assessed by
physiological measurements combined with global expression profiling
(proteomics). Parallel experiments were carried out using the NahG transgenic
plants expressing the bacterial gene encoding SA hydroxylase, which cannot
accumulate the active form of this plant defense elicitor. SA markedly improved
germination under salt stress. Proteomic analyses disclosed a specific
accumulation of protein spots regulated by SA as inferred by silver-nitrate
staining of two-dimensional gels, detection of carbonylated (oxidized) proteins,
and neosynthesized proteins with [35S]-methionine. The combined results revealed
several processes potentially affected by SA. This molecule enhanced the
reinduction of the late maturation program during early stages of germination,
thereby allowing the germinating seeds to reinforce their capacity to mount
adaptive responses in environmental water stress. Other processes affected by SA
concerned the quality of protein translation, the priming of seed metabolism, the
synthesis of antioxidant enzymes, and the mobilization of seed storage proteins.
All the observed effects are likely to improve seed vigor. Another aspect
revealed by this study concerned the oxidative stress entailed by SA in
germinating seeds, as inferred from a characterization of the carbonylated
(oxidized) proteome. Finally, the proteomic data revealed a close interplay
between abscisic signaling and SA elicitation of seed vigor.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16679420 [PubMed - indexed for MEDLINE]

1336: Plant Cell Rep. 2006 Oct;25(10):1035-42. Epub 2006 May 3.

Investigation of the endosperm-specific sucrose synthase promoter from rice using
transient expression of reporter genes in guar seed tissue.

Rasmussen TB, Donaldson IA.

Danisco Innovation, Langebrogade 1, 1001, Copenhagen K, Denmark.

We report the investigation of an endosperm-specific promoter from the rsus3 gene
from rice (Oryza sativa). The promoter was characterized by deletion analysis and
transient expression in guar (Cyamopsis tetragonoloba) seed-tissue. Transient
expression was monitored by histochemical GUS assay, and quantitative dual
reporter assays comprising firefly luciferase as a test reporter, and Renilla
luciferase and GUS as reference reporters. These revealed high expression levels
of the reporter genes directed by the rsus3 promoter in guar endosperm.
Specificity for this tissue in seeds was apparent from a virtual absence of
reporter activity in guar cotyledons. Removal of a putative intron region only
slightly raised the expression level, whereas duplication of the minimal promoter
region, in a tandem-repeat rsus3 promoter construct, retained endosperm
specificity in guar, and displayed three times the reporter activity observed
with the single copy construct.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16670901 [PubMed - indexed for MEDLINE]

1337: Genetics. 2006 Jul;173(3):1621-8. Epub 2006 Apr 30.

Fitness benefits of systemic acquired resistance during Hyaloperonospora
parasitica infection in Arabidopsis thaliana.

Heidel AJ, Dong X.

Department of Biology, Duke University, Durham, North Carolina 27708, USA.
aheidel@fli-leibniz.de

We investigated the fitness benefits of systemic acquired resistance (SAR) in
Arabidopsis thaliana using a mutational and transformational genetic approach.
Genetic lines were designed to differ in the genes determining resistance
signaling in a common genetic background. Two mutant lines (cpr1 and cpr5)
constitutively activate SAR at different points in SAR signaling, and one mutant
line (npr1) has impaired SAR. The transgenic line (NPR1-H) has enhanced
resistance when SAR is activated, but SAR is still inducible similarly to wild
type. The fitness benefits were also investigated under two nutrient levels to
test theories that preventing pathogen damage and realized resistance benefits
may be affected by nutrient availability. Under low-nutrient conditions and
treatment with the pathogenic oomycete, Hyaloperonospora parasitica, wild type
had a higher fitness than the mutant that could not activate SAR, demonstrating
that normal inducible SAR is beneficial in these conditions; this result,
however, was not found under high-nutrient conditions. The mutants with
constitutive SAR all failed to show a fitness benefit in comparison to wild type
under a H. parasitica pathogen treatment, suggesting that SAR is induced to
prevent an excessive fitness cost.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16648642 [PubMed - indexed for MEDLINE]

1338: J Biotechnol. 2006 Oct 1;125(4):447-61. Epub 2006 Apr 27.

The effect of polysaccharide-degrading wine yeast transformants on the efficiency
of wine processing and wine flavour.

Louw C, La Grange D, Pretorius IS, van Rensburg P.

Institute for Wine Biotechnology, Department of Viticulture and Oenology,
Stellenbosch University, Matieland (Stellenbosch), ZA 7602, South Africa.

Commercial polysaccharase preparations are applied to winemaking to improve wine
processing and quality. Expression of polysaccharase-encoding genes in
Saccharomyces cerevisiae allows for the recombinant strains to degrade
polysaccharides that traditional commercial yeast strains cannot. In this study,
we constructed recombinant wine yeast strains that were able to degrade the
problem-causing grape polysaccharides, glucan and xylan, by separately
integrating the Trichoderma reesei XYN2 xylanase gene construct and the
Butyrivibrio fibrisolvens END1 glucanase gene cassette into the genome of the
commercial wine yeast strain S. cerevisiae VIN13. These genes were also combined
in S. cerevisiae VIN13 under the control of different promoters. The strains that
were constructed were compared under winemaking conditions with each other and
with a recombinant wine yeast strain expressing the endo-beta-1,4-glucanase gene
cassette (END1) from B. fibrisolvens and the endo-beta-1,4-xylanase gene cassette
(XYN4) from Aspergillus niger, a recombinant strain expressing the pectate lyase
gene cassette (PEL5) from Erwinia chrysanthemi and the polygalacturonase-encoding
gene cassette (PEH1) from Erwinia carotovora. Wine was made with the recombinant
strains using different grape cultivars. Fermentations with the recombinant VIN13
strains resulted in significant increases in free-flow wine when Ruby Cabernet
must was fermented. After 6 months of bottle ageing significant differences in
colour intensity and colour stability could be detected in Pinot Noir and Ruby
Cabernet wines fermented with different recombinant strains. After this period
the volatile composition of Muscat d'Alexandria, Ruby Cabernet and Pinot Noir
wines fermented with different recombinant strains also showed significant
differences. The Pinot Noir wines were also sensorial evaluated and the tasting
panel preferred the wines fermented with the recombinant strains.

Publication Types:
Evaluation Studies

PMID: 16644051 [PubMed - indexed for MEDLINE]

1339: Plant Cell Rep. 2006 Oct;25(10):1111-21. Epub 2006 Apr 27.

Overexpression of carnation S-adenosylmethionine decarboxylase gene generates a
broad-spectrum tolerance to abiotic stresses in transgenic tobacco plants.

Wi SJ, Kim WT, Park KY.

Department of Biology, College of Natural Science, Sunchon National University,
Sunchon, Chonnam 540-742, Korea.

Polyamines (PAs), such as putrescine, spermidine, and spermine, are present in
all living organism and implicate in a wide range of cellular physiological
processes. We have used transgenic technology in an attempt to evaluate their
potential for mitigating the adverse effects of several abiotic stresses in
plants. Sense construct of full-length cDNA for S-adenosylmethionine
decarboxylase (SAMDC), a key enzyme in PA biosynthesis, from carnation (Dianthus
caryophyllus L.) flower was introduced into tobacco (Nicotiana tabacum L.) by
Agrobacterium tumefaciens-mediated transformation. Several transgenic lines
overexpressing SAMDC gene under the control of cauliflower mosaic virus 35S
promoter accumulated soluble total PAs by 2.2 (S16-S-4) to 3.1 (S16-S-1) times
than wild-type plants. The transgenic tobacco did not show any difference in
organ phenotype compared to the wild-type. The number and weight of seeds
increased, and net photosynthetic rate also increased in transgenic plants.
Stress-induced damage was attenuated in these transgenic plants, in the symptom
of visible yellowing and chlorophyll degradation after all experienced stresses
such as salt stress, cold stress, acidic stress, and abscisic acid treatment.
H2O2-induced damage was attenuated by spermidine treatment. Transcripts for
antioxidant enzymes (ascorbate peroxidase, manganase superoxide dismutase, and
glutathione S-transferase) in transgenic plants and GUS activity transformed with
SAMDC promoter::GUS fusion were induced more significantly by stress treatment,
compared to control. These results that the transgenic plants with sense SAMDC
cDNA are more tolerant to abiotic stresses than wild-type plants suggest that PAs
may play an important role in contributing stress tolerance in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16642382 [PubMed - indexed for MEDLINE]

1340: Appl Microbiol Biotechnol. 2006 Aug;71(5):598-607. Epub 2006 Apr 26.

Genetically modified crops: success, safety assessment, and public concern.

Singh OV, Ghai S, Paul D, Jain RK.

Department of Pediatrics, The John Hopkins School of Medicine, Baltimore, MD
21287, USA. osingh1@jhmi.edu

With the emergence of transgenic technologies, new ways to improve the agronomic
performance of crops for food, feed, and processing applications have been
devised. In addition, ability to express foreign genes using transgenic
technologies has opened up options for producing large quantities of commercially
important industrial or pharmaceutical products in plants. Despite this high
adoption rate and future promises, there is a multitude of concerns about the
impact of genetically modified (GM) crops on the environment. Potential
contamination of the environment and food chains has prompted detailed
consideration of how such crops and the molecules that they produce can be
effectively isolated and contained. One of the reasonable steps after creating a
transgenic plant is to evaluate its potential benefits and risks to the
environment and these should be compared to those generated by traditional
agricultural practices. The precautionary approach in risk management of GM
plants may make it necessary to monitor significant wild and weed populations
that might be affected by transgene escape. Effective risk assessment and
monitoring mechanisms are the basic prerequisites of any legal framework to
adequately address the risks and watch out for new risks. Several agencies in
different countries monitor the release of GM organisms or frame guidelines for
the appropriate application of recombinant organisms in agro-industries so as to
assure the safe use of recombinant organisms and to achieve sound overall
development. We feel that it is important to establish an internationally
harmonized framework for the safe handling of recombinant DNA organisms within a
few years.

Publication Types:
Review

PMID: 16639559 [PubMed - indexed for MEDLINE]

1341: Plant Physiol. 2006 Jun;141(2):578-86. Epub 2006 Apr 21.

The intracellular fate of a recombinant protein is tissue dependent.

Drakakaki G, Marcel S, Arcalis E, Altmann F, Gonzalez-Melendi P, Fischer R,
Christou P, Stoger E.

Institute for Molecular Biotechnology, Biology VII, Aachen University, 52074
Aachen, Germany.

Recombinant proteins directed to the secretory pathway in plants require a signal
peptide for entry into the endoplasmic reticulum. In the absence of further
targeting information, such proteins are generally secreted via the default
pathway to the apoplast. This has been well documented in protoplasts and leaf
tissue, but the trafficking of recombinant proteins in seeds and other storage
tissues has rarely been investigated. We used Aspergillus niger phytase as a
model glycoprotein to compare the intracellular fate of a recombinant protein in
the leaves and seeds of rice (Oryza sativa). Using fluorescence and electron
microscopy we showed that the recombinant protein was efficiently secreted from
leaf cells as expected. In contrast, within endosperm cells it was retained in
endoplasmic reticulum-derived prolamin bodies and protein storage vacuoles.
Consistent with our immunolocalization data, the phytase produced in endosperm
cells possessed oligomannose and vacuolar-type N-glycans
[Man(3)(Xyl)(Fuc)GlcNAc(2)], whereas the phytase produced in leaves contained
predominantly secretion-type N-glycans [GlcNAc(2)Man(3)(Xyl)(Fuc)GlcNAc(2)]. The
latter could not be detected in preparations of the endosperm-derived phytase.
Our results show that the intracellular deposition and modification of a
recombinant protein is tissue dependent.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16632592 [PubMed - indexed for MEDLINE]

1342: Pest Manag Sci. 2006 Jun;62(6):558-64.

Metabolism of [14C]-2,4-dichlorophenol in edible plants.

Laurent F, Debrauwer L, Pascal-Lorber S.

INRA, UMR Xö©nobiotiques, 180 Ch. de Tournefeuille, BP3, F-31931 Toulouse Cedex 9,
France. flaurent@toulouse.inra.fr

Several 2,4-dichlorophenoxyacetic acid (2,4-D)-sensitive plants have been
modified by genetic engineering with tfdA gene to acquire 2,4-D tolerance. The
expression product of this gene degrades 2,4-D to 2,4-dichlorophenol (DCP), which
is less phytotoxic but could cause a problem of food safety. After a comparison
of 2,4-D and DCP metabolism in transgenic 2,4-D-tolerant and wild cotton
(Gossypium hirsutum L.), a direct study of DCP metabolism in edible plants was
performed. After petiolar uptake of a [U-phenyl-(14)C]-DCP solution followed by a
48 h water chase, aqueous extracts were analysed by high-performance liquid
chromatography. Metabolites were thereafter isolated and their structural
identities were determined by enzymatic and chemical hydrolyses and mass
spectrometry analyses. The metabolic fate of DCP was equivalent to 2,4-D
metabolism in transgenic 2,4-D-tolerant cotton. In addition, DCP metabolism was
similar in transgenic and wild cotton. The major terminal metabolites were
DCP-saccharide conjugates in all species, essentially DCP-(6-O-malonyl)-glucoside
or its precursor DCP-glucose. The significance of this metabolic pathway with
regard to food safety is discussed. Copyright (c) 2006 Society of Chemical
Industry

Publication Types:
Comparative Study

PMID: 16628540 [PubMed - indexed for MEDLINE]

1343: Metab Eng. 2006 Jul;8(4):291-302. Epub 2006 Apr 18.

Understanding carotenoid metabolism as a necessity for genetic engineering of
crop plants.

Sandmann G, Rö¶mer S, Fraser PD.

Molecular Biosciences 213, P.O. Box 111932, J. W. Goethe Universitö¤t, D-60054
Frankfurt, Germany. sandmann@em.uni-frankfurt.de

As a proof of concept, the qualitative and quantitative engineering of carotenoid
formation has been achieved in crop plants. Successful reports in tomato, potato,
rice, and canola all describe the enhancement of carotenoid with nutritional
value, while in model systems such as tobacco and Arabidopsis the engineering of
carotenoid to confer abiotic stress has been described. For all the successful
applications there have been many examples of unintended/unpredicted phenotypes
and results. Typically this has resided from our lack of understanding of
carotenoid formation and its regulation. In the present article, we will review
advances in carotenoid formation and its regulation to illustrate how metabolic
engineering experiments have shed light on regulatory mechanisms.

Publication Types:
Review

PMID: 16621640 [PubMed - indexed for MEDLINE]

1344: J Biotechnol. 2006 Sep 18;125(3):361-8. Epub 2006 Apr 18.

Automated DNA extraction from genetically modified maize using
aminosilane-modified bacterial magnetic particles.

Ota H, Lim TK, Tanaka T, Yoshino T, Harada M, Matsunaga T.

Malcom Co., Ltd.,4-15-10 Honmachi Shibuya-ku, Tokyo 151-0071, Japan.

A novel, automated system, PNE-1080, equipped with eight automated pestle units
and a spectrophotometer was developed for genomic DNA extraction from maize using
aminosilane-modified bacterial magnetic particles (BMPs). The use of
aminosilane-modified BMPs allowed highly accurate DNA recovery. The
(A(260)-A(320)):(A(280)-A(320)) ratio of the extracted DNA was 1.9+/-0.1. The DNA
quality was sufficiently pure for PCR analysis. The PNE-1080 offered rapid assay
completion (30 min) with high accuracy. Furthermore, the results of real-time PCR
confirmed that our proposed method permitted the accurate determination of
genetically modified DNA composition and correlated well with results obtained by
conventional cetyltrimethylammonium bromide (CTAB)-based methods.

Publication Types:
Evaluation Studies

PMID: 16621089 [PubMed - indexed for MEDLINE]

1345: Phytochemistry. 2006 Aug;67(16):1750-7. Epub 2006 Apr 17.

Metabolite profiling of carotenoid and phenolic pathways in mutant and transgenic
lines of tomato: identification of a high antioxidant fruit line.

Long M, Millar DJ, Kimura Y, Donovan G, Rees J, Fraser PD, Bramley PM, Bolwell
GP.

School of Biological Sciences, Royal Holloway, University of London, Egham,
Surrey TW20 0EX, UK.

Plant secondary metabolism is highly regulated within the major pathways to
terpenoids, phenolics and alkaloids. Such regulation can occur at multiple levels
from transcription through to the compartmentation of the product. However, the
possibility exists for cross-talk between these pathways, the regulation of which
is largely unknown at present. Such phenomena are important to understand in the
application of plant breeding, where unintended effects of transgenesis or
mutation can have an impact on the environment or human health. In an effort to
improve dietary antioxidant content of crop plants, the tomato has been a major
focus of effort for engineering both lipophilic antioxidants such as carotenoids
and hydrophilic antioxidants such as flavonoid glycosides. In this study, a panel
of transgenic and mutant tomato lines has been subjected to metabolite profiling
in comparison with wild type Ailsa Craig for both carotenoids and phenolics. A
range of mutants and transgenic lines were selected showing a range of phenotypes
varying from down-regulation through to increased levels of lycopene and
beta-carotene. All mutants altered in structural genes for carotenoid
biosynthesis showed that perturbations in carotenoid biosynthesis do not
generally alter phenolic or flavonoids content significantly even when devoid of
carotenoids. Reciprocally, the down-regulation of ferulate 5-hydroxylase had no
effect on carotenoid content. In contrast mutants defective in light perception
such as the high pigment (hp-1) and LA3771 possess elevated chlorogenic acid and
rutin as well as increased carotenoid content. These lines can act as the hosts
for further genetic manipulation for increased antioxidant content.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16616263 [PubMed - indexed for MEDLINE]

1346: Plant Cell Physiol. 2006 Jun;47(6):756-63. Epub 2006 Apr 13.

The correlation between expression and localization of a foreign gene product in
rice endosperm.

Yasuda H, Hayashi Y, Jomori T, Takaiwa F.

Department of Plant Biotechnology, National institute of Agrobiological Sciences,
Tsukuba, Ibaraki, 305-8602 Japan.

Glucagon-like peptide 1 (GLP-1) is a 30 amino acid peptide hormone involved in
insulin stimulation that is dependent upon blood glucose levels. We have
previously reported that when this short peptide gene was directly expressed
under the control of a glutelin promoter and its signal peptide, it was not
accumulated in transgenic rice seed due to gene silencing. However, when the
modified GLP-1 (mGLP-1) gene was enlarged to 5xmGLP-1 (mGLPx5) by tandem repeat,
no silencing was observed. The mGLPx5 peptide could be accumulated in rice seed
and its localization was mainly limited to the endoplasmic reticulum (ER). We
also investigated alternative cellular localization sites that would increase
accumulation. The relationship between the expression level and localization was
examined by attaching the chitinase signal peptide to mGLPx5 to direct it into
the intercellular space (apoplast), or by expression as a fusion protein with
glutelin by insertion into a variable region of the acidic subunit, thus
directing the peptide to protein body II (PB II). Attachment of the KDEL ER
retention signal to the 6xmGLP-1 (mGLPx6) or its fusion to the C-terminus of the
13 kDa prolamin directed the peptide to the ER or PB I, respectively.
Unexpectedly, these results indicated that mGLPx5 without any signal except for
the glutelin signal peptide was accumulated to the greatest extent in rice
endosperm. It can thus be concluded that the ER is a suitable intracellular
organelle for accumulation of mGLPx5 peptide.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16614094 [PubMed - indexed for MEDLINE]

1347: Plant Cell Rep. 2006 Sep;25(9):920-6. Epub 2006 Apr 12.

Comparative analysis of 35S and lectin promoters in transgenic soybean tissue
using an automated image acquisition system and image analysis.

Buenrostro-Nava MT, Ling PP, Finer JJ.

Department of Horticulture and Crop Science, The Ohio State University/OARDC,
Wooster, OH 44691, USA.

Expression of the green fluorescent protein (gfp) gene, under regulatory control
of either the constitutive 35S promoter or the developmentally-regulated lectin
promoter was monitored and quantified using a newly-developed automated tracking
system. The automated system consisted of a computer-controlled two-dimensional
robotics table and a programmable image acquisition system, which was used to
semi-continuously monitor gfp gene expression during development of transgenic
soybean [Glycine max (L.) Merrill] somatic embryos. Quantitative analysis of GFP
expression showed that, during somatic embryo proliferation and early
development, expression of lectin-GFP was not detected. During late embryo
development, expression of lectin-GFP gradually increased until the levels were
similar to those of 35S-GFP. The use of an automated image collection system and
image analysis facilitated the frequent monitoring and quantification of gfp gene
expression on a large number of samples over an extended period of time.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16609890 [PubMed - indexed for MEDLINE]

1348: Diabetologia. 2006 Jun;49(6):1324-32. Epub 2006 Mar 29.

Apolipoprotein AV does not contribute to hypertriglyceridaemia or triglyceride
lowering by dietary fish oil and rosiglitazone in obese Zucker rats.

Dorfmeister B, Brandlhofer S, Schaap FG, Hermann M, Fö¼rnsinn C, Hagerty BP,
Stangl H, Patsch W, Strobl W.

Department of Medical Chemistry, Center of Physiology and Pathophysiology,
Medical University of Vienna, Wö¤hringer Strasse 9, A-1090 Vienna, Austria.

AIMS/HYPOTHESIS: Apolipoprotein AV (apoAV) is a recently discovered
apolipoprotein with a triglyceride-lowering effect in genetically modified mice.
Transcription of the human gene encoding apoAV (APOA5) is suppressed by insulin
and stimulated by fibrates. Our goal was to study the expression of Apoa5, in
comparison with Apoa4 and Apoc3, in hypertriglyceridaemic, obese and
insulin-resistant Zucker rats receiving the insulin sensitiser rosiglitazone
and/or a fish oil diet to lower triglycerides. METHODS: Hepatic Apoa5, Apoa4 and
Apo3 mRNA and liver and plasma apoAV were measured in lean and obese Zucker rats
receiving rosiglitazone while on a coconut oil or fish oil diet. RESULTS: Basal
hepatic Apoa5 expression was similar in obese and lean Zucker rats. Unexpectedly,
obese Zucker rats tended to have higher plasma apoAV levels despite their
hypertriglyceridaemic state. Both rosiglitazone and the fish oil diet
significantly increased Apoa5 mRNA, by about 70%, but tended to lower liver and
plasma apoAV. Rosiglitazone had no effect on Apoa5 mRNA in cultured rat
hepatocytes. No intact PPAR (peroxisome proliferator-activated receptor) response
element was identified in the rat Apoa5 promoter. CONCLUSIONS/INTERPRETATION: Our
data indicate that apoAV does not contribute to the hypertriglyceridaemia of
obese Zucker rats or to the hypolipidaemic effect of rosiglitazone or a fish oil
diet. The divergent changes of Apoa5 mRNA and apoAV levels suggest co- or
post-translational regulation. The increase in Apoa5 mRNA induced by
rosiglitazone is not directly mediated by peroxisome proliferator-activated
receptor gamma.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16570166 [PubMed - indexed for MEDLINE]

1349: Plant Cell Rep. 2006 Sep;25(9):942-52. Epub 2006 Mar 25.

Stability of transgene integration and expression in subsequent generations of
doubled haploid oilseed rape transformed with chitinase and beta-1,3-glucanase
genes in a double-gene construct.

Melander M, Kamnert I, Happstadius I, Liljeroth E, Bryngelsson T.

Svalö¶f Weibull AB, 268 81, Svalö¶v, Sweden. margareta.melander@plantscience.se

A double-gene construct with one chitinase and one beta-1,3-glucanase gene from
barley, both driven by enhanced 35S promoters, was transformed into oilseed rape.
From six primary transformants expressing both transgenes 10 doubled haploid
lines were produced and studied for five generations. The number of inserted
copies for both the genes was determined by Southern blotting and real-time PCR
with full agreement between the two methods. When copy numbers were analysed in
different generations, discrepancies were found, indicating that at least part of
the inserted sequences were lost in one of the alleles of some doubled haploids.
Chitinase and beta-1,3-glucanase expression was analysed by Western blotting in
all five doubled haploid generations. Despite that both the genes were present on
the same T-DNA and directed by the same promoter their expression pattern between
generations was different. The beta-1,3-glucanase was expressed at high and
stable levels in all generations, while the chitinase displayed lower expression
that varied between generations. The transgenic plants did not show any major
impact on fungal resistance when assayed in greenhouse, although purified
beta-1,3-glucanase and chitinase caused retardment of fungal growth in vitro.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16565860 [PubMed - indexed for MEDLINE]

1350: Plant Cell Rep. 2006 Sep;25(9):953-8. Epub 2006 Mar 25.

Mannose selection system used for cucumber transformation.

He Z, Duan Z, Liang W, Chen F, Yao W, Liang H, Yue C, Sun Z, Chen F, Dai J.

Biotechnology Research Center, China Three Gorges University, Yichang 443002, PR
China.

The selectable marker system, which utilizes the pmi gene encoding for
phosphomannose-isomerase that converts mannose-6-phosphate to
fructose-6-phosphate, was adapted for Agrobacterium-mediated transformation of
cucumber (Cucumis sativus L.). Only transformed cells were capable of utilizing
mannose as a carbon source. The highest transformation frequency of 23% was
obtained with 10 g/l mannose and 10 g/l sucrose in the medium. Molecular, genetic
analysis, and PMI activity assay showed that the regenerated shoots contained the
pmi gene and the gene was transmitted to the progeny in a Mendelian fashion. The
results indicated that the mannose selection system, which is devoid of the
disadvantages of antibiotic or herbicide selection, could be used for cucumber
Agrobacterium-mediated transformation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16565859 [PubMed - indexed for MEDLINE]

1351: Plant Cell Rep. 2006 Sep;25(9):936-41. Epub 2006 Mar 22.

Anther-specific expression of mutated melon ethylene receptor gene Cm-ERS1/H70A
affected tapetum degeneration and pollen grain production in transgenic tobacco
plants.

Takada K, Ishimaru K, Kamada H, Ezura H.

Gene Research Center, University of Tsukuba, Ten-nodai 1-1-1, Tsukuba 305-8572,
Japan.

To develop a new system for inducible male sterility without any modification of
the floral architecture in tobacco plants, a mutated ethylene receptor gene
Cm-ERS1/H70A was fused either to the tobacco Nin88 promoter known to function
mainly in the tapetum and microspore or to the CaMV 35S promoter known to be a
constitutive promoter. The fusion genes pNin88::Cm-ERS1/H70A and
p35S::Cm-ERS1/H70A were introduced in tobacco plants, which generated two
independent transformants. Transformants with 35S::Cm-ERS1/H70A produced less
normal pollen and had modified floral architecture while those with
Nin88::Cm-ERS1/H70A produced less normal pollen without modification of floral
architecture. Histological observations of anthers at stage 2 showed that tapetum
degeneration in NH70A #8 and H70A #2 transformants occurred later than in wild
types, strongly indicating that the expression of the mutated gene was involved
in this delay. These results suggest that the tapetum-specific expression of a
mutated ethylene receptor gene is a potential strategy for inducing male
sterility in transgenic plants.

PMID: 16552596 [PubMed - indexed for MEDLINE]

1352: Planta. 2006 Aug;224(3):598-611. Epub 2006 Mar 22.

Structure, expression, and functional analysis of the hexokinase gene family in
rice (Oryza sativa L.).

Cho JI, Ryoo N, Ko S, Lee SK, Lee J, Jung KH, Lee YH, Bhoo SH, Winderickx J, An
G, Hahn TR, Jeon JS.

Plant Metabolism Research Center & Graduate School of Biotechnology, Kyung Hee
University, 449-701 Yongin, Republic of South Korea.

Hexokinase (HXK) is a dual-function enzyme that both phosphorylates hexose to
form hexose 6-phosphate and plays an important role in sugar sensing and
signaling. To investigate the roles of hexokinases in rice growth and
development, we analyzed rice sequence databases and isolated ten rice hexokinase
cDNAs, OsHXK1 (Oryza sativa Hexokinase 1) through OsHXK10. With the exception of
the single-exon gene OsHXK1, the OsHXKs all have a highly conserved genomic
structure consisting of nine exons and eight introns. Gene expression profiling
revealed that OsHXK2 through OsHXK9 are expressed ubiquitously in various organs,
whereas OsHXK10 expression is pollen-specific. Sugars induced the expression of
three OsHXKs, OsHXK2, OsHXK5, and OsHXK6, in excised leaves, while suppressing
OsHXK7 expression in excised leaves and immature seeds. The hexokinase activity
of the OsHXKs was confirmed by functional complementation of the
hexokinase-deficient yeast strain YSH7.4-3C (hxk1, hxk2, glk1). OsHXK4 was able
to complement this mutant only after the chloroplast-transit peptide was removed.
The subcellular localization of OsHXK4 and OsHXK7, observed with green
fluorescent protein (GFP) fusion constructs, indicated that OsHXK4 is a
plastid-stroma-targeted hexokinase while OsHXK7 localizes to the cytosol.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16552590 [PubMed - indexed for MEDLINE]

1353: Plant Cell Rep. 2006 Aug;25(8):799-806. Epub 2006 Mar 14.

Strategies to obtain stable transgenic plants from non-embryogenic lines:
complementation of the nn1 mutation of the NORK gene in Medicago sativa MN1008.

Perhald A, Endre G, Kevei Z, Kiss GB, Kereszt A.

Institute of Genetics, Biological Research Center of the Hungarian Academy of
Sciences, Temesvari korut 62, 6726, Szeged, Hungary.

Strategies to introduce genes into non-embryogenic plants for complementation of
a mutation are described and tested on tetraploid alfalfa (Medicago sativa).
Genes conditioning embryogenic potential, a mutant phenotype, and a gene to
complement the mutation can be combined using several different crossing and
selection steps. In the successful strategy used here, the M. sativa genotype
MnNC-1008(NN) carrying the recessive non-nodulating mutant allele nn ( 1 ) was
crossed with the highly embryogenic alfalfa line Regen S and embryogenic hybrid
individuals were identified from the F1 progeny. After transformation of these
hybrids with the wild-type gene (NORK), an F2 generation segregating for the
mutation and transgene were produced. Plants homozygous for the mutant allele and
carrying the wild-type NORK transgene could form root nodules after inoculation
with Sinorhizobium meliloti demonstrating successful complementation of the nn (
1 ) mutation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16534599 [PubMed - indexed for MEDLINE]

1354: Plant Cell Rep. 2006 Aug;25(8):815-20. Epub 2006 Mar 10.

Molecular analysis of transgene and vector backbone integration into the barley
genome following Agrobacterium-mediated transformation.

Lange M, Vincze E, Mö¸ller MG, Holm PB.

Department of Genetics and Biotechnology, Danish Institute of Agricultural
Sciences, Research Centre Flakkebjerg, Forsoegsvej 1, DK-4200, Slagelse, Denmark.
mette.lange@agrsci.dk

We report a large-scale study on the frequency of transgene and T-DNA backbone
integration following Agrobacterium-mediated transformation of immature barley
embryos. One hundred and ninety-one plant lines were regenerated after hygromycin
selection and visual selection for GFP expression at the callus stage. Southern
blotting performed on a subset of 53 lines that were PCR positive for the GFP
gene documented the integration of the GFP gene in 27 of the lines. Twenty-three
of these lines expressed GFP in T(1) plantlets. Southern blotting with a vector
backbone probe revealed that 13 of the 27 lines possessed one or more vector
backbone fragments illustrating the regular occurrence of vector backbone
integration following Agrobacterium infection of barley immature embryos.

PMID: 16528561 [PubMed - indexed for MEDLINE]

1355: Plant Cell Rep. 2006 Jul;25(7):651-9. Epub 2006 Mar 2.

A high throughput Agrobacterium tumefaciens-mediated transformation method for
functional genomics of perennial ryegrass (Lolium perenne L.).

Bajaj S, Ran Y, Phillips J, Kularajathevan G, Pal S, Cohen D, Elborough K,
Puthigae S.

HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand.
sbajaj@hortresearch.co.nz

A robust and high throughput Agrobacterium genetic transformation procedure has
been developed for perennial ryegrass (Lolium perenne L.). Embryogenic callus
lines were selected and maintained as plants in vitro. Embryogenic calli derived
from meristematic regions of the vegetative tillers were co-cultivated with
Agrobacterium tumefaciens strain EHA101 carrying the plasmid pCAMBIA 1305.1 in
the presence of acetosyringone for 3-4 days. The calli were grown under 94.8 and
151.6 microM hygromycin selection, respectively for two cycles of 2-weeks each,
followed by transfer to regeneration medium with 47.4 microM hygromycin.
Regenerated plants were rooted and successfully transferred to soil. The
transgenic nature of the regenerated plants was confirmed by DNA gel blot
analysis and gene expression demonstrated by GUS histochemical assay and/or
reverse transcription PCR. After development of the transformation procedure, we
used Agrobacterium strain EHA101 carrying a modified binary plasmid pMH bearing
genes of interest. In the past 2 years, we have produced more than 1,000 plants
with constructs encoding different genes of interest from perennial ryegrass.

PMID: 16518636 [PubMed - indexed for MEDLINE]

1356: Environ Pollut. 2006 Nov;144(1):70-6. Epub 2006 Mar 2.

Overexpressing both ATP sulfurylase and selenocysteine methyltransferase enhances
selenium phytoremediation traits in Indian mustard.

LeDuc DL, AbdelSamie M, Mö¨ntes-Bayon M, Wu CP, Reisinger SJ, Terry N.

Department of Plant and Microbial Biology, University of California, Berkeley, CA
94720-3012, USA.

A major goal of our selenium (Se) phytoremediation research is to use genetic
engineering to develop fast-growing plants with an increased ability to tolerate,
accumulate, and volatilize Se. To this end we incorporated a gene (encoding
selenocysteine methyltransferase, SMT) from the Se hyperaccumulator, Astragalus
bisulcatus, into Indian mustard (LeDuc, D.L., Tarun, A.S., Montes-Bayö¨n, M.,
Meija, J., Malit, M.F., Wu, C.P., AbdelSamie, M., Chiang, C.-Y., Tagmount, A.,
deSouza, M., Neuhierl, B., Bö¶ck, A., Caruso, J., Terry, N., 2004. Overexpression
of selenocysteine methyltransferase in Arabidopsis and Indian mustard increases
selenium tolerance and accumulation Plant Physiol. 135, 377-383.). The resulting
transgenic plants successfully enhanced Se phytoremediation in that the plants
tolerated and accumulated Se from selenite significantly better than wild type.
However, the advantage conferred by the SMT enzyme was much less when Se was
supplied as selenate. In order to enhance the phytoremediation of selenate, we
developed double transgenic plants that overexpressed the gene encoding ATP
sulfurylase (APS) in addition to SMT, i.e., APSxSMT. The results showed that
there was a substantial improvement in Se accumulation from selenate (4 to 9
times increase) in transgenic plants overexpressing both APS and SMT.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16515825 [PubMed - indexed for MEDLINE]

1357: J Plant Physiol. 2007 Feb;164(2):126-36. Epub 2006 Mar 2.

Overexpression of CaHSP26 in transgenic tobacco alleviates photoinhibition of
PSII and PSI during chilling stress under low irradiance.

Guo SJ, Zhou HY, Zhang XS, Li XG, Meng QW.

College of Life Sciences, Shandong Agricultural University, Tai'an, PR China.

A sweet pepper cDNA clone, CaHSP26 encoding the chloroplast (CP)-localized small
heat shock protein (sHSP), was isolated and characterized with regard to its
sequence, response to various temperatures and function in transgenic tobacco
plants. The deduced amino acid sequence contained three highly conserved regions,
showing high identities with other plant sHSPs. Expression of the CaHSP26 gene
showed that the mRNA accumulation of CaHSP26 was induced by heat stress. Higher
transcript levels were observed when sweet pepper leaves were treated at 42
degrees C for 3h. However, the expression of the CaHSP26 gene was not induced by
chilling stress (4 degrees C) in the absence of heat shock (HS). But the
transcripts were still detected at 48h at 4 degrees C after HS while not at 25
degrees C. The photochemical efficiency of PSII (Fv/Fm) and the oxidizable P700
in transgenic tobacco overexpressing CaHSP26 were higher than that in wild type
tobacco during chilling stress under low irradiance. These results suggest that
CP sHSP protein plays an important role in protection of PSII and PSI during
chilling stress under low irradiance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16513207 [PubMed - indexed for MEDLINE]

1358: Ecotoxicol Environ Saf. 2007 Feb;66(2):195-203. Epub 2006 Feb 24.

Occurrence and persistence of Bacillus thuringiensis (Bt) and transgenic Bt corn
cry1Ab gene from an aquatic environment.

Douville M, Gagnö© F, Blaise C, Andrö© C.

Environment Canada, St. Lawrence Centre, 105 McGill Street, Montrö©al, Quö©.,
Canada H2Y 2E7.

Genetically modified corn crops and suspensions of Bacillus thuringiensis (Bt)
are currently used to control pest infestations of insects of the Lepidoptera
family. For this purpose, the cry1Ab gene coding for protein delta-endotoxin
derived from B. thuringiensis kurstaki (Btk), which is highly toxic to these
insects, was inserted and expressed in corn. The aims of this study were to
examine the occurrence and persistence of the cry1Ab gene from Btk and Bt corn in
aquatic environments near fields where Bt corn was cultivated. First, an optimal
DNA preparation and extraction methodology was developed to allow for
quantitative gene analysis by real-time polymerase chain reaction (qPCR) in
various environmental matrices. Second, surface water and sediment were spiked in
vitro with genomic DNA from Bt or Bt corn to evaluate the persistence of cry1Ab
genes. Third, soil, sediment, and water samples were collected before seeding, 2
weeks after pollen release, and after corn harvesting and mechanical root
remixing in soils to assess cry1Ab gene content. DNA was extracted with
sufficient purity (i.e., low absorbance at 230 nm and absence of PCR-inhibiting
substances) from soil, sediment, and surface water. The cry1Ab gene persisted for
more than 21 and 40 days in surface water and sediment, respectively. The removal
of bacteria by filtration of surface water samples did not significantly increase
the half-life of the transgene, but the levels were fivefold more abundant than
those in unfiltered water at the end of the exposure period. In sediments, the
cry1Ab gene from Bt corn was still detected after 40 days in clay- and sand-rich
sediments. Field surveys revealed that the cry1Ab gene from transgenic corn and
from naturally occurring Bt was more abundant in the sediment than in the surface
water. The cry1Ab transgene was detected as far away as the Richelieu and St.
Lawrence rivers (82 km downstream from the corn cultivation plot), suggesting
that there were multiple sources of this gene and/or that it undergoes transport
by the water column. Sediment-associated cry1Ab gene from Bt corn tended to
decrease with distance from the Bt cornfield. Sediment concentrations of the
cry1Ab gene were significantly correlated with those of the cry1Ab gene in
surface water (R=0.83;P=0.04). The data indicate that DNA from Bt corn and Bt
were persistent in aquatic environments and were detected in rivers draining
farming areas.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16499967 [PubMed - indexed for MEDLINE]

1359: J Virol Methods. 2006 Jun;134(1-2):217-22. Epub 2006 Feb 20.

Expression of avian reovirus sigmaC protein in transgenic plants.

Huang LK, Liao SC, Chang CC, Liu HJ.

Institute of Biotechnology, National Cheng Kung University, Tainan 701, Taiwan.

Avian reovirus (ARV) structural protein, sigmaC encoded by S1 genome segment, is
the prime candidate to become a vaccine against ARV infection. Two plant nuclear
expression vectors with expression of sigmaC-encoding gene driven by CaMV 35S
promoter and rice actin promoter were constructed, respectively. Agrobacterium
containing the S1 expression constructs were used to transform alfalfa, and
transformants were selected using hygromysin. The integration of S1 transgene in
alfalfa chromosome was confirmed by PCR and histochemical GUS staining. Western
blot analysis using antiserum against sigmaC was carried out to determine the
expression of sigmaC protein in transgenic alfalfa cells. The highest expression
levels of sigmaC protein in the cellular extracts of selected p35S-S1 and
pAct1-S1 transgenic alfalfa lines were 0.008% and 0.007% of the total soluble
protein, respectively. The transgenic alfalfa cells with expression of sigmaC
protein pave the way for the development of edible vaccine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16488486 [PubMed - indexed for MEDLINE]

1360: Food Chem Toxicol. 2006 Jul;44(7):1092-9. Epub 2006 Feb 17.

Results of a 90-day safety assurance study with rats fed grain from corn
borer-protected corn.

Hammond BG, Dudek R, Lemen JK, Nemeth MA.

Monsanto Company, Product Safety Center, 800 North Lindbergh Blvd., Bldg. O3F,
St. Louis, MO 63167, USA. bruce.g.hammond@monsanto.com

The results of a 90-day rat feeding study with grain from MON 810 corn (YieldGard
Cornborer -- YieldGard Cornborer is a registered trademark of Monsanto
Technology, LLC) that is protected against feeding damage from corn and stalk
boring lepidopteran insects are presented. Corn borer protection was accomplished
through the introduction of cry1Ab coding sequences into the corn genome for in
planta production of a bioactive form of Cry1Ab protein. Grain from MON 810 and
its near-isogenic control was separately formulated into rodent diets at levels
of 11% and 33% (w/w) by Purina Mills, Inc. (PMI). All diets were nutritionally
balanced and conformed to PMI specifications for Certified LabDiet (PMI Certified
LabDiet 5002 is a registered trademark of Purina Mills, Inc.) 5002. There were a
total of 400 rats in the study divided into 10 groups of 20 rats/sex/group. The
responses of rats fed diets containing MON 810 were compared to those of rats fed
grain from conventional corn varieties. Overall health, body weight, food
consumption, clinical pathology parameters (hematology, blood chemistry,
urinalysis), organ weights, and gross and microscopic appearance of tissues were
comparable between groups fed diets containing MON 810 and conventional corn
varieties. This study complements extensive agronomic, compositional and farm
animal feeding studies with MON 810 grain, confirming that it is as safe and
nutritious as grain from existing commercial corn varieties.

PMID: 16487643 [PubMed - indexed for MEDLINE]

1361: Planta. 2006 Aug;224(3):622-32. Epub 2006 Feb 16.

Regulation of carotenoid and ABA accumulation during the development and
germination of Nicotiana plumbaginifolia seeds.

Frey A, Boutin JP, Sotta B, Mercier R, Marion-Poll A.

INRA-INAPG, Institut Jean-Pierre Bourgin, UMR204, Laboratoire de Biologie des
Semences, 78026 Versailles Cedex, France.

Abscisic acid (ABA) is derived from epoxycarotenoid cleavage and regulates seed
development and maturation. A detailed carotenoid analysis was undertaken to
study the contribution of epoxycarotenoid synthesis to the regulation of ABA
accumulation in Nicotiana plumbaginifolia developing seeds. Maximal accumulation
of xanthophylls occurred at mid-development in wild type seeds, when total ABA
levels also peaked. In contrast, in ABA-deficient mutants xanthophyll synthesis
was delayed, in agreement with the retardation in seed maturation. Seed dormancy
was restored in mutants impaired in the conversion of zeaxanthin into
violaxanthin by zeaxanthin epoxidase (ZEP), by the introduction of the
Arabidopsis AtZEP gene under the control of promoters inducing expression during
later stages of seed development compared to wild type NpZEP, and in dry and
imbibed seeds. Alterations in the timing and level of ZEP expression did not
highly affect the temporal regulation of ABA accumulation in transgenic seeds,
despite notable perturbations in xanthophyll accumulation. Therefore, major
regulatory control of ABA accumulation might occur downstream of epoxycarotenoid
synthesis.

PMID: 16482436 [PubMed - indexed for MEDLINE]

1362: Environ Pollut. 2006 Oct;143(3):449-55. Epub 2006 Feb 3.

Cry1Ab protein from Bt transgenic rice does not residue in rhizosphere soil.

Wang H, Ye Q, Wang W, Wu L, Wu W.

Institute of Nuclear Agricultural Sciences, Zhejiang University, Hua-jia-chi
Campus, Hangzhou 310029, China.

Expression of Cry1Ab protein in Bt transgenic rice (KMD) and its residue in the
rhizosphere soil during the whole growth in field, as well as degradation of the
protein from KMD straw in five soils under laboratory incubation were studied.
The residue of Cry1Ab protein in KMD rhizosphere soil was undetectable (below the
limit of 0.5 ng/g air-dried soil). The Cry1Ab protein contents in the shoot and
root of KMD were 3.23-8.22 and 0.68-0.89 microg/g (fresh weight), respectively.
The half-lives of the Cry1Ab protein in the soils amended with KMD straw (4%,
w/w) ranged from 11.5 to 34.3d. The residence time of the protein varied
significantly in a Fluvio-marine yellow loamy soil amended with KMD straw at the
rate of 3, 4 and 7%, with half-lives of 9.9, 13.8 and 18d, respectively. In
addition, an extraction method for Cry1Ab protein in soil was developed, with
extraction efficiencies of 46.4-82.3%.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16459002 [PubMed - indexed for MEDLINE]

1363: Planta. 2006 Jul;224(2):347-59. Epub 2006 Feb 1.

Isolation of a cDNA clone (PcSrp) encoding serine-rich-protein from Porteresia
coarctata T. and its expression in yeast and finger millet (Eleusine coracana L.)
affording salt tolerance.

Mahalakshmi S, Christopher GS, Reddy TP, Rao KV, Reddy VD.

Centre for Plant Molecular Biology, Osmania University, 500 007 Hyderabad, AP,
India.

A 1.4 Kb cDNA clone encoding a serine-rich protein has been isolated from the
cDNA library of salt stressed roots of Porteresia coarctata, and designated as P.
coarctata serine-rich-protein (PcSrp) encoding gene. Northern analysis and in
situ mRNA hybridization revealed the expression of PcSrp in the salt stressed
roots and rhizome of P. coarctata. However, no such expression was seen in the
salt stressed leaves and in the unstressed tissues of root, rhizome and leaf,
indicating that PcSrp is under the control of a salt-inducible tissue-specific
promoter. In yeast, the PcSrp conferred increased NaCl tolerance, implicating its
role in salinity tolerance at cellular level. Further, PcSrp was cloned
downstream to rice Actin-1 promoter and introduced into finger millet through
particle-inflow-gun method. Transgenic plants expressing PcSrp were able to grow
to maturity and set seed under 250 mM NaCl stress. The untransformed control
plants by contrast failed to survive under similar salt stress. The stressed
roots of transgenic plants invariably accumulated higher Na(+) and K(+) ion
contents compared to roots of untransformed plants; whereas, shoots of
transgenics accumulated lower levels of both the ions than that of untransformed
plants under identical stress, clearly suggesting the involvement of PcSrp in ion
homeostasis contributing to salt tolerance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16450172 [PubMed - indexed for MEDLINE]

1364: Planta. 2006 Jul;224(2):405-12. Epub 2006 Feb 1.

Characterization and functional analysis of a pollen-specific gene st901 in
Solanum tuberosum.

Zhao Y, Zhao Q, Ao G, Yu J.

State Key Laboratory for Agricultural Biotechnology, College of Biological
Sciences, China Agricultural University, No. 2 Yuanmingyuan West Road, 100094,
Beijing, People's Republic of China.

A pollen-specific gene, sb401, which was isolated from a cDNA library of in vitro
geminated pollen of the diploid potato species Solanum berthaultii, belongs to
the class of genes expressed late during pollen development. Using sb401 as a
probe, a pollen-specific gene st901 was isolated from the genomic library of a
potato species Solanum tuberosum cv. Desiree. Sequencing and RT-PCR analysis
showed that the st901 genomic gene is 2,889 bp long, contains three exons and two
introns, and encodes a putative polypeptide of 217 residues. The predicted
protein sequence contains four imperfect repeated motifs of V-V-E-K-K-N/E-E; the
core sequence of the repeats (K-K-N/E-E) resembles a microtubule-binding domain
of the microtubule-associated protein MAP1B from mouse. The examination of a
promoter-reporter construct in transgenic potato plants revealed that the st901
is expressed exclusively in mature pollen grains, which is consistent with the
results of Northern blot and RT-PCR. For analysis of the function of st901,
transgenic plants harboring antisense copies of st901 cDNA driven by a native
st901 promoter were generated. Suppression of st901 gene in potato resulted in
aberrant pollen at maturation and pollen viability of transgenic plants ranged
from 4.4 to 14.8%, while that of control plants were more than 90%. These results
strongly suggest that st901 has an essential role in pollen development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16450169 [PubMed - indexed for MEDLINE]

1365: Food Chem Toxicol. 2006 Jul;44(7):964-73. Epub 2006 Jan 19.

Sub-chronic (13-week) oral toxicity study in rats with recombinant human
lactoferrin produced in the milk of transgenic cows.

Appel MJ, van Veen HA, Vietsch H, Salaheddine M, Nuijens JH, Ziere B, de Loos F.

TNO Quality of Life, Business Unit Toxicology and Applied Pharmacology, P.O. Box
360, 3700 AJ Zeist, The Netherlands. appel@voeding.tno.nl

The oral toxicity of recombinant human lactoferrin (rhLF) produced in the milk of
transgenic cows was investigated in Wistar rats by daily administration via oral
gavage for 13 consecutive weeks, 7 days per week. The study used four groups of
20 rats/sex/dose. The control group received physiological saline and the three
test groups received daily doses of 200, 600 and 2000 mg of rhLF per kg body
weight. Clinical observations, growth, food consumption, food conversion
efficiency, water consumption, neurobehavioural testing, ophthalmoscopy,
haematology, clinical chemistry, renal concentration test, urinalysis, organ
weights and gross examination at necropsy and microscopic examination of various
organs and tissues were used as criteria for detecting the effects of treatment.
Overall, no treatment-related, toxicologically significant changes were observed.
The few findings that may be related to the treatment (lower cholesterol in
high-dose females, lower urinary pH in high-dose males and females and very
slightly higher kidney weight in high-dose females) were considered of no
toxicological significance. Based on the absence of treatment-related,
toxicologically relevant changes, the no-observed-adverse-effect level (NOAEL)
was considered to be at least 2000 mg/kg body weight/day.

PMID: 16426723 [PubMed - indexed for MEDLINE]

1366: Plant Cell Rep. 2006 Jun;25(6):554-60. Epub 2006 Jan 18.

Bacillus thuringiensis protein production, signal transduction, and insect
control in chemically inducible PR-1a/cry1Ab broccoli plants.

Cao J, Bates SL, Zhao JZ, Shelton AM, Earle ED.

Department of Plant Breeding and Genetics, Cornell University, Ithaca, NY 14853,
USA.

In an effort to develop a chemically inducible system for insect management, we
studied production of Cry1Ab Bacillus thuringiensis (Bt) protein and control of
the diamondback moth (DBM), Plutella xylostella L., in inducer-treated and
untreated tissues of a broccoli line transformed with a PR-1a/cry1Ab expression
cassette. Spraying leaves of these plants with the inducer acibenzolar-S-methyl
(= 1,2,3 benzothiadiazole-7-thiocarboxylic acid-S-methyl-ester) (ASM) triggered
expression of the cry1Ab gene and produced a high level of Cry1Ab protein within
2-3 days. Cry1Ab protein persisted in leaves for at least 8 weeks, providing
prolonged protection from P. xylostella attack. Signals generated in
inducer-treated leaves were transferred to untreated newly emerged leaves or
heads, as seen by production of Cry1Ab protein and/or protection from insect
damage in these plant parts. Signal transduction proceeded in an attenuated
manner up to the sixth newly emerged leaf. No Cry1Ab protein was detectable by
ELISA in uninduced young leaves, but small amounts of the protein were present in
uninduced leaves older than 3 weeks and caused some insect mortality. Such basal
expression of Bt genes without induction may favor the evolution of resistant
insect populations and therefore limits the application of the PR-1a/cry1Ab
system for insect management. However, the rapid production and steady
maintenance of a high level of transgenic protein upon induction, the signal
transduction observed, and the fact that the chemical inducer can be used in
field conditions make the PR-1a promoter attractive for chemical regulation of
other agriculturally or pharmaceutically important genes for which low expression
in the absence of induction is not a concern.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16418860 [PubMed - indexed for MEDLINE]

1367: Plant Cell Rep. 2006 Jun;25(6):573-81. Epub 2006 Jan 12.

Tobacco ribosomal DNA spacer element elevates Bowman-Birk inhibitor expression in
tomato plants.

Yakoby N, Garvey A, Raskin I.

Biotech Center, Cook College, Rutgers University, Foran Hall, 59 Dudley Road, New
Brunswick, NJ 08901-8520, USA. nyakoby@Princeton.edu

Amplification promoting sequence (aps), from tobacco rDNA, was found to induce
amplification and enhances the expression of heterologous genes, consequently
increasing the expression of transgenic proteins in tobacco. In this report we
demonstrate that aps element also affects integration, transcription, and
translation of a soybean protease inhibitor, Bowman-Birk inhibitor (BBI), in
transgenic tomato plants and quantifies its effects in different expression
vectors. A synthetic bbi gene was constructed, based on the wild-type gene
containing two independent inhibition sites; trypsin and chymotrypsin.
Transformation vectors were designed using two different promoters; the tomato
fruit specific E8 promoter and the constitutively active 35S CaMV promoter. These
vectors were transformed into 'Moneymaker' tomato plants. In tomato fruits and
leaves, aps caused a 3-fold increase in bbi mRNA levels when compared to the
lines without aps. Similar increases were obtained in plants expressing bbi
controlled by E8 or 35S CaMV promoters. Also, the level of BBI protein expression
in aps-transformed plants was 3 fold-higher than in plants without aps. This is
the first report of aps effect on the enhanced gene expression and transgenic
protein production in plant other than tobacco.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16408179 [PubMed - indexed for MEDLINE]

1368: Planta. 2006 Jul;224(2):300-14. Epub 2006 Jan 6.

Rice ascorbate peroxidase gene family encodes functionally diverse isoforms
localized in different subcellular compartments.

Teixeira FK, Menezes-Benavente L, Galvö¸o VC, Margis R, Margis-Pinheiro M.

Laboratö¨rio de Genö©tica Molecular Vegetal, Departamento de Genö©tica, Universidade
Federal do Rio de Janeiro, 21944-970, Rio de Janeiro, Brasil.

Aerobic organisms evolved a complex antioxidant system, which protect the cells
against oxidative damage caused by partially reduced oxygen intermediates, also
known as reactive oxygen species. In plants, ascorbate peroxidases (EC,
1.11.1.11) catalyze the conversion of H(2)O(2) to H(2)O, using ascorbate as the
specific electron donor in this enzymatic reaction. Previously, eight APx genes
were identified in the rice (Oryza sativa L.) genome through in silico analysis:
two cytosolic isoforms, two putative peroxisomal isoforms, and four putative
chloroplastic ones. Using gene-specific probes, we confirmed the presence of the
eight APx genes in the rice genome by Southern blot hybridization. Transcript
accumulation analysis showed specific expression patterns for each member of the
APx family according to developmental stage and in response to salt stress,
revealing the complexity of the antioxidant system in plants. Finally, the
subcellular localization of rice APx isoforms was determined using GFP-fusion
proteins in BY-2 tobacco cells. In agreement with the initial prediction, OSAPX3
was localized in the peroxisomes. On the other hand, the OSAPX6-GFP fusion
protein was found in mitochondria of the BY-2 cells, in contrast to the
chloroplastic location predicted by sequence analysis. Our findings reveal the
functional diversity of the rice APx genes and suggest complementation and
coordination of the antioxidant defenses in different cellular compartments
during development and abiotic stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16397796 [PubMed - indexed for MEDLINE]

1369: Planta. 2006 Jun;224(1):125-32. Epub 2006 Jan 4.

RNAi-mediated silencing of the myo-inositol-1-phosphate synthase gene (GmMIPS1)
in transgenic soybean inhibited seed development and reduced phytate content.

Nunes AC, Vianna GR, Cuneo F, Amaya-Farfö¡n J, de Capdeville G, Rech EL, Aragö¸o
FJ.

Embrapa Recursos Genö©ticos e Biotecnologia, PqEB W5 Norte, 70770-900 Brasö­lia,
DF, Brazil.

Inositol plays a role in membrane trafficking and signaling in addition to
regulating cellular metabolism and controlling growth. In plants, the
myo-inositol-1-phosphate is synthesized from glucose 6-phosphate in a reaction
catalyzed by the enzyme myo-inositol-1-phosphate synthase (EC 5.5.1.4). Inositol
can be converted into phytic acid (phytate), the most abundant form of phosphate
in seeds. The path to phytate has been suggested to proceed via the sequential
phosphorylation of inositol phosphates, and/or in part via phosphatidylinositol
phosphate. Soybean [Glycine max (L.) Merrill] lines were produced using
interfering RNA (RNAi) construct in order to silence the myo-inositol-1-phosphate
(GmMIPS1) gene. We have observed an absence of seed development in lines in which
the presence of GmMIPS1 transcripts was not detected. In addition, a drastic
reduction of phytate (InsP6) content was achieved in transgenic lines (up to
94.5%). Our results demonstrated an important correlation between GmMIPS1 gene
expression and seed development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16395584 [PubMed - indexed for MEDLINE]

1370: Planta. 2006 Jun;224(1):155-62. Epub 2006 Jan 4.

Expression of a plant expansin is involved in the establishment of root knot
nematode parasitism in tomato.

Gal TZ, Aussenberg ER, Burdman S, Kapulnik Y, Koltai H.

Department of Genomics, ARO, The Volcani Center, 50250 Bet Dagan, Israel.

A group of plant proteins, expansins, have been identified as wall-loosening
factors and as facilitators of cell expansion in vivo. The root knot nematode
Meloidogyne javanica establishes a permanent feeding site composed of giant cells
surrounded by gall tissue. We used quantitative PCR and in situ localization to
demonstrate the induction of a tomato (Lycopersicon esculentum cv. VF36) expansin
(LeEXPA5) expression in gall cells adjacent to the nematode feeding cells. To
further characterize the biological role of LeEXPA5 we have generated
LeEXPA5-antisense transgenic roots. The ability of the nematode to establish a
feeding site and complete its life cycle, the average root cell size and the rate
of root elongation were determined for the transgenic roots, as well as the level
of LeEXPA5 expression in non-infected and nematode-infected roots. Our results
demonstrated that a decrease of LeEXPA5 expression reduces the ability of the
nematode to complete its life cycle in transgenic roots. We suggest that a
plant-originated expansin is necessary for a successful parasitic nematode-plant
interaction.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16395582 [PubMed - indexed for MEDLINE]

1371: Pharmacol Ther. 2006 Aug;111(2):374-83. Epub 2005 Dec 20.

Genetically modified plants and food hypersensitivity diseases: usage and
implications of experimental models for risk assessment.

Prescott VE, Hogan SP.

Division of Molecular Bioscience, The John Curtin School of Medical Research,
Australian National University, Canberra, ACT, Australia.

The recent advances in biotechnology in the plant industry have led to increasing
crop production and yield that in turn has increased the usage of genetically
modified (GM) food in the human food chain. The usage of GM foods for human
consumption has raised a number of fundamental questions including the ability of
GM foods to elicit potentially harmful immunological responses, including
allergic hypersensitivity. To assess the safety of foods derived from GM plants
including allergenic potential, the US FDA, Food and Agriculture Organization of
the United Nations (FAO)/World Health Organization (WHO), and the EU have
developed approaches for evaluation assessment. One assessment approach that has
been a very active area of research and debate is the development and usage of
animal models to assess the potential allergenicity of GM foods. A number of
specific animal models employing rodents, pigs, and dogs have been developed for
allergenicity assessment. However, validation of these models is needed and
consideration of the criteria for an appropriate animal model for the assessment
of allergenicity in GM plants is required. We have recently employed a BALB/c
mouse model to assess the potential allergenicity of GM plants. We have been able
to demonstrate that this model is able to detect differences in antigenicity and
identify aspects of protein post-translational modifications that can alter
antigenicity. Furthermore, this model has also enabled us to examine the usage of
GM plants as a therapeutic approach for the treatment of allergic diseases. This
review discusses the current approaches to assess the allergenic potential of GM
food and particularly focusing on the usage of animal models to determine the
potential allergenicity of GM foods and gives an overview of our recent findings
and implications of these studies.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 16364445 [PubMed - indexed for MEDLINE]

1372: Peptides. 2006 Jun;27(6):1179-86. Epub 2005 Dec 13.

Design of genetically modified soybean proglycinin A1aB1b with multiple copies of
bioactive peptide sequences.

Prak K, Maruyama Y, Maruyama N, Utsumi S.

Laboratory of Food Quality Design and Development, Graduate School of
Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.

The peptide IIAEK derived from beta-lactoglobulin has a hypocholesterolemic
activity greater than that of beta-sitosterol. To create food proteins with
multiple copies of this valuable peptide sequence, we introduced tandem multimers
of the nucleotide sequence encoding the peptide into DNA regions corresponding to
the five variable regions of soybean glycinin A1aB1b subunit, and expressed the
mutants in Escherichia coli. The expression level and solubility of the five
mutants, each containing four IIAEK sequences in each of the variable regions,
were compared. Overall, the expression level and solubility of the mutants with
four IIAEK sequences in the variable regions IV and V were the best followed by
II > III > I. Further, introduction of the fifth IIAEK sequence to the variable
region IV did not decrease expression level and solubility. Increasing the number
of IIAEK to 7 and 10 slightly decreased expression level, while their solubility
decreased to as low as 40 and 1%, respectively. Various mutations were combined
to get a mutant containing as many IIAEK sequences as possible. Some of the
resulting mutants were expressed in the soluble form. The mutant containing eight
IIAEK from the combination of variable regions IV and V (IV-4 + V-4) showed the
best balance of the expression level and solubility, followed by the combination
of variable regions II and III (II-4 + III-4). The soluble fractions of these
mutants were purified by hydrophobic, gel filtration and ion-exchange column
chromatography. Yields of IIAEK peptide released by in vitro digestion with
trypsin from both mutants were around 80%. This is the first report that a large
amount of a physiologically active peptide could be introduced into food protein.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16356590 [PubMed - indexed for MEDLINE]

1373: J Plant Physiol. 2007 Jan;164(1):23-30. Epub 2005 Dec 13.

Oleate accumulation, induced by silencing of microsomal omega-6 desaturase,
declines with leaf expansion in transgenic tobacco.

Yang M, Xu Y.

Key Laboratory of Photosynthesis and Environmental Molecular Physiology,
Institute of Botany, Chinese Academy of Sciences, Nanxincun 20, Xiangshan,
Beijing 100093, China.

All higher plants contain at least one microsomal omega-6 desaturase (FAD2),
which inserts a double bond between the carbons 12 and 13 of monounsaturated
oleic acid to generate polyunsaturated linoleic acid and controls most of the
polyunsaturated lipid synthesis in plant cells. RNA interference can be used to
silence endogenous genes by effective degradation of target transcripts. To
investigate development-related silencing of the FAD2, fatty acid composition was
analyzed in the context of leaf expansion in FAD2-silenced tobacco lines obtained
by RNA interference technology. We observed that the increased oleate level in
unexpanded leaves due to FAD2-silencing receded significantly in fully expanded
leaves. The mechanism involved in this interesting phenomenon was investigated by
analyses of individual lipid proportion, fatty acid composition of individual
lipids, and FAD2 transcript level in the transgenic leaves at different expansion
stages. Data revealed that the expansion-related FAD2-silencing effect was not
due to rebound of FAD2 transcript, but rather probably due to chloroplast
development with leaf expansion.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16352373 [PubMed - indexed for MEDLINE]

1374: Bioresour Technol. 2006 Dec;97(18):2345-9. Epub 2005 Dec 2.

Optimization of recombinant hyperthermophilic esterase production from
agricultural waste using response surface methodology.

Ren X, Yu D, Han S, Feng Y.

Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education,
Jilin University, Changchun, PR China.

The aim of this work was to evaluate the capability of corn steep liquor being a
low cost substrate on the recombinant protein production by cultivating
recombinant Escherichia coli and increasing the production of hyperthermophilic
esterase (HE). The effect of corn steep liquor, mineral salt and trace metals on
hyperthermophilic esterase production was investigated by means of a five-level
three-factor central composite rotatable design. Optimized values of the factors
were determined and a maximum hyperthermophilic esterase production of 251.39
U/ml was obtained. This value equaled the yield by yeast extract and peptone
medium on the whole.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16330210 [PubMed - indexed for MEDLINE]

1375: Planta. 2006 Jun;224(1):32-41. Epub 2005 Dec 2.

BcXTH1, a Brassica campestris homologue of Arabidopsis XTH9, is associated with
cell expansion.

Shin YK, Yum H, Kim ES, Cho H, Gothandam KM, Hyun J, Chung YY.

School of Life Sciences and Biotechnology, Korea University, 136-701 Seoul,
Anam-Dong, Korea.

Xyloglucan endotransglucosylase/hydrolases (XTHs) are a group of the enzymes that
are responsible for reorganization of the cellulose-xyloglucan framework by
catalyzing cleavage and religation of the xyloglucan chains in the plant cell
wall. In this study, we report the isolation and characterization of a XTH gene
from a pistil cDNA library of Brassica campestris. Sequence analysis of the gene,
designated BcXTH1, revealed that it is homologous to the XTH9 gene of
Arabidopsis. The highly conserved domain (DEIDFEFLG) found among all XTHs was
also present in BcXTH1 but with the two amino acid substitutions (NEFDFEFLG) also
found in Arabidopsis XTH9. These results suggest that BcXTH1 is the B. campestris
homologue of XTH9. Expression analysis of BcXTH1 revealed that it was expressed
in most of the plant organs. In situ hybridization showed that the gene is highly
expressed in the floral primodia, especially in the epidermal cell layer.
Southern blot analysis indicated that the BcXTH1 gene exists as a multi-copy gene
in the B. campestris genome. The function of the BcXTH1 gene was deduced from
using an overexpression strategy in Arabidopsis. Interestingly, the transgenic
plants showed a pronounced cell expansion phenotype. Immunoelectron microscopy
shows that BcXTH1 is localized almost exclusively to the cell wall, supporting
our conclusion that it participates in the regulation of cell expansion in B.
campestris.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16322981 [PubMed - indexed for MEDLINE]

1376: Environ Pollut. 2006 Jul;142(2):212-6. Epub 2005 Nov 28.

Consequences for Protaphorura armata (Collembola: Onychiuridae) following
exposure to genetically modified Bacillus thuringiensis (Bt) maize and non-Bt
maize.

Heckmann LH, Griffiths BS, Caul S, Thompson J, Pusztai-Carey M, Moar WJ, Andersen
MN, Krogh PH.

National Environmental Research Institute, Department of Terrestrial Ecology,
Vejlsö¸vej 25, PO Box 314, DK-8600 Silkeborg, Denmark.

Studies on the effect of genetically modified Bacillus thuringiensis (Bt) crops
on true soil dwelling non-target arthropods are scarce. The objective of this
study was to assess the influence of a 4-week exposure to two Bt maize varieties
(Cry1Ab) Cascade and MEB307 on the collembolan Protaphorura armata. For
comparison three non-Bt maize varieties, Rivaldo (isogenic to Cascade),
Monumental (isogenic to MEB307) and DK242, and two control diets based on baker's
yeast (uncontaminated and contaminated with Bt toxin Cry1Ab) were also tested.
Due to a lower C:N ratio, individuals reared on yeast performed significantly
better in all of the measured endpoints than those reared on maize. P. armata
performed equally well when reared on two Bt and three non-Bt maize varieties.
Although there were no negative effects of Bt maize in this experiment, we
recommend future studies on Bt crops to focus on species interactions in
long-term, multi-species experiments.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16310913 [PubMed - indexed for MEDLINE]

1377: Theriogenology. 2006 Jun;65(9):1800-12. Epub 2005 Nov 21.

An approach for producing transgenic cloned cows by nuclear transfer of cells
transfected with human alpha 1-antitrypsin gene.

Jang G, Bhuiyan MM, Jeon HY, Ko KH, Park HJ, Kim MK, Kim JJ, Kang SK, Lee BC,
Hwang WS.

Department of Therigenology and Biotechnology, College of Veterinary Medicine,
Seoul National University, San56-1, Kwanak-Gu, Seoul 151-742, South Korea.

In an attempt to produce transgenic cloned cows secreting alpha 1-antitrypsin
(alpha1-AT) protein into milk, bovine cumulus cells were transfected with a
plasmid containing an alpha1-AT gene and green fluorescent protein (GFP) reporter
gene using Fugene 6 as a lipid carrier. The GFP-expressing cells were selected
and transferred into enucleated bovine oocytes. Couplets were fused, chemically
activated and cultured. Developmental competence was monitored and the number of
inner cell mass (ICM) and trophectoderm (TE) cells in blastocysts were counted
after differential staining. The percentage of blastocysts was lower (P < 0.05)
in transgenic cloned embryos compared to non-transgenic cloned embryos (23%
versus 35%). No difference in the numbers of ICM and TE cells between the two
groups of embryos was observed. One or two GFP-expressing blastocysts were
transferred into the uterus of each recipient cow. Out of 49 recipient cows,
three pregnancies were detected by non-return estrus and rectal palpation.
However, the pregnancies failed to maintain to term; two fetuses were aborted at
Day 60 and 150, respectively, and one fetus at Day 240. The genomic DNA from the
aborted fetus was amplified by polymerase chain reaction (PCR) to investigate
integration of the transgene in the fetus. The expected PCR product was sequenced
and was identical to the sequence of alpha1-AT transgene. In conclusion, the
present study demonstrated that developmental competence of cloned embryos
derived from transgenic donor cells was lower than embryos derived from
non-transfected donor cells. Although we failed to obtain a viable transgenic
cloned calf, integration of alpha1-AT gene into the fetus presents the
possibility of producing transgenic cloned cows by somatic cell nuclear transfer.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16303172 [PubMed - indexed for MEDLINE]

1378: Phytochemistry. 2006 Aug;67(15):1590-7. Epub 2005 Nov 18.

The primary in vivo steroidal alkaloid glucosyltransferase from potato.

McCue KF, Allen PV, Shepherd LV, Blake A, Whitworth J, Maccree MM, Rockhold DR,
Stewart D, Davies HV, Belknap WR.

USDA, Agricultural Research Service, Crop Improvement and Utilization Research
Unit, Western Regional Research Center, 800 Buchanan St., Albany, CA 94710-1105,
USA.

To provide tools for breeders to control the steroidal glycoalkaloid (SGA)
pathway in potato, we have investigated the steroidal alkaloid
glycosyltransferase (Sgt) gene family. The committed step in the SGA pathway is
the glycosylation of solanidine by either UDP-glucose or UDP-galactose leading to
alpha-chaconine or alpha-solanine, respectively. The Sgt2 gene was identified by
deduced protein sequence homology to the previously identified Sgt1 gene. SGT1
has glucosyltransferase activity in vitro, but in vivo serves as the
UDP-galactose:solanidine galactosyltransferase. Two alleles of the Sgt2 gene were
isolated and its function was established with antisense transgenic lines and in
vitro assays of recombinant protein. In tubers of transgenic potato (Solanum
tuberosum) cvs. Lenape and Desirö©e expressing an antisense Sgt2 gene construct,
accumulation of alpha-solanine was increased and alpha-chaconine was reduced.
Studies with recombinant SGT2 protein purified from yeast show that SGT2
glycosylation activity is highly specific for UDP-glucose as a sugar donor. This
data establishes the function of the gene product (SGT2), as the primary
UDP-glucose:solanidine glucosyltransferase in vivo.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16298403 [PubMed - indexed for MEDLINE]