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.
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.
Fei
Houman
H
Department of Plant Science, University of Manitoba, Winnipeg, MB, Canada R3T 2N2.
Chaillou
Sylvain
S
Hirel
Bertrand
B
Polowick
Patricia
P
Mahon
John D
JD
Vessey
J Kevin
JK
D018533:Biomass; D003600:Cytosol / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D005974:Glutamate-Ammonia Ligase / Q000235:genetics / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D009928:Organ Specificity; D018532:Pisum sativum / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism* / Q000494:pharmacology; D013025:Glycine max / Q000166:cytology / Q000201:enzymology*
D000644:Quaternary Ammonium Compounds; D005974:Glutamate-Ammonia Ligase; D009584:Nitrogen