Overexpression of cytosolic glutamine synthetase. Relation to nitrogen, light, and photorespiration.
In plants, ammonium released during photorespiration exceeds primary nitrogen assimilation by as much as 10-fold. Analysis of photorespiratory mutants indicates that photorespiratory ammonium released in mitochondria is reassimilated in the chloroplast by a chloroplastic isoenzyme of glutamine synthetase (GS2), the predominant GS isoform in leaves of Solanaceous species including tobacco (Nicotiana tabacum). By contrast, cytosolic GS1 is expressed in the vasculature of several species including tobacco. Here, we report the effects on growth and photorespiration of overexpressing a cytosolic GS1 isoenzyme in leaf mesophyll cells of tobacco. The plants, which ectopically overexpress cytosolic GS1 in leaves, display a light-dependent improved growth phenotype under nitrogen-limiting and nitrogen-non-limiting conditions. Improved growth was evidenced by increases in fresh weight, dry weight, and leaf soluble protein. Because the improved growth phenotype was dependent on light, this suggested that the ectopic expression of cytosolic GS1 in leaves may act via photosynthetic/photorespiratory process. The ectopic overexpression of cytosolic GS1 in tobacco leaves resulted in a 6- to 7-fold decrease in levels of free ammonium in leaves. Thus, the overexpression of cytosolic GS1 in leaf mesophyll cells seems to provide an alternate route to chloroplastic GS2 for the assimilation of photorespiratory ammonium. The cytosolic GS1 transgenic plants also exhibit an increase in the CO(2) photorespiratory burst and an increase in levels of photorespiratory intermediates, suggesting changes in photorespiration. Because the GS1 transgenic plants have an unaltered CO(2) compensation point, this may reflect an accompanying increase in photosynthetic capacity. Together, these results provide new insights into the possible mechanisms responsible for the improved growth phenotype of cytosolic GS1 overexpressing plants. Our studies provide further support for the notion that the ectopic overexpression of genes for cytosolic GS1 can potentially be used to affect increases in nitrogen use efficiency in transgenic crop plants.
Department of Biology, New York University, NY 10003, USA.
Igor C
IC
Oliveira
Timothy
T
Brears
Thomas J
TJ
Knight
Alexandra
A
Clark
Gloria M
GM
Coruzzi
D002245:Carbon Dioxide / Q000378:metabolism / Q000528:radiation effects; D003600:Cytosol / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000528:radiation effects; D005974:Glutamate-Ammonia Ligase / Q000235:genetics* / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008027:Light; D009154:Mutation; D009584:Nitrogen / Q000494:pharmacology*; D010641:Phenotype; D010788:Photosynthesis / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics; D030821:Plants, Genetically Modified; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics*
D007527:Isoenzymes; D000644:Quaternary Ammonium Compounds; D002245:Carbon Dioxide; D005974:Glutamate-Ammonia Ligase; D009584:Nitrogen