title: Assimilation of excess ammonium into amino acids and nitrogen translocation in Arabidopsis thaliana--roles of glutamate synthases and carbamoylphosphate synthetase in leaves.
abstract: This study was aimed at investigating the physiological role of ferredoxin-glutamate synthases (EC 1.4.1.7), NADH-glutamate synthase (EC 1.4.1.14) and carbamoylphosphate synthetase (EC 6.3.5.5) in Arabidopsis. Phenotypic analysis revealed a high level of photorespiratory ammonium, glutamine/glutamate and asparagine/aspartate in the GLU1 mutant lacking the major ferredoxin-glutamate synthase, indicating that excess photorespiratory ammonium was detoxified into amino acids for transport out of the veins. Consistent with these results, promoter analysis and in situ hybridization demonstrated that GLU1 and GLU2 were expressed in the mesophyll and phloem companion cell-sieve element complex. However, these phenotypic changes were not detected in the GLU2 mutant defective in the second ferredoxin-glutamate synthase gene. The impairment in primary ammonium assimilation in the GLT mutant under nonphotorespiratory high-CO(2) conditions underlined the importance of NADH-glutamate synthase for amino acid trafficking, given that this gene only accounted for 3% of total glutamate synthase activity. The excess ammonium from either endogenous photorespiration or the exogenous medium was shifted to arginine. The promoter analysis and slight effects on overall arginine synthesis in the T-DNA insertion mutant in the single carbamoylphosphate synthetase large subunit gene indicated that carbamoylphosphate synthetase located in the chloroplasts was not limiting for ammonium assimilation into arginine. The data provided evidence that ferredoxin-glutamate synthases, NADH-glutamate synthase and carbamoylphosphate synthetase play specific physiological roles in ammonium assimilation in the mesophyll and phloem for the synthesis and transport of glutamine, glutamate, arginine, and derived amino acids.
authors: Unité de Nutrition Azotée des Plantes, Institut National de la Recherche Agronomique, Versailles, France.
authors: Fabien
authors: F
authors: Potel
authors: Marie-Hélène
authors: MH
authors: Valadier
authors: Sylvie
authors: S
authors: Ferrario-Méry
authors: Olivier
authors: O
authors: Grandjean
authors: Halima
authors: H
authors: Morin
authors: Laure
authors: L
authors: Gaufichon
authors: Stéphanie
authors: S
authors: Boutet-Mercey
authors: Jérémy
authors: J
authors: Lothier
authors: Steven J
authors: SJ
authors: Rothstein
authors: Naoya
authors: N
authors: Hirose
authors: Akira
authors: A
authors: Suzuki
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D001692:Biological Transport; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D050790:Glutamate Synthase (NADH) / Q000235:genetics* / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D009586:Nitrogen Fixation; D018515:Plant Leaves / Q000201:enzymology*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D004269:DNA, Bacterial; D018744:DNA, Plant; D000644:Quaternary Ammonium Compounds; C036483:T-DNA; D050790:Glutamate Synthase (NADH); D002222:Carbamoyl-Phosphate Synthase (Ammonia); D009584:Nitrogen