Analysis of glutamate homeostasis by overexpression of Fd-GOGAT gene in Arabidopsis thaliana.
Glutamate plays a central role in nitrogen flow and serves as a nitrogen donor for the production of amino acids. In plants, some amino acids work as buffers: during photorespiration, ammonium derived from the conversion of glycine to serine is promptly reassimilated into glutamate by the glutamine synthetase (GS-2)/ferredoxin-dependent glutamate synthase (Fd-GOGAT) cycle. The glutamate concentration is relatively stable compared with those of other amino acids under environmental changes. The few studies dealing with glutamate homeostasis have but all used knockouts or mutants of these enzymes. Here, we generated Fd-GOGAT (GLU1)-overexpressing Arabidopsis plants to analyze changes in the amino acid pool caused by glutamate overproduction under different ammonium conditions controlled by CO(2) concentration, light intensity and nitrate concentration. Under photorespiratory conditions with sufficient ammonium supply, aspartate increased and glutamine and glycine decreased, but glutamate barely changed. Under non-photorespiratory conditions, however, glutamate and most other amino acids increased. These results suggest that the synthesized glutamate is promptly converted into other amino acids, especially aspartate. In addition, ammonium supply by photorespiration does not limit glutamate biosynthesis, but glutamine and glycine are important. This study will contribute to the understanding of glutamate homeostasis in plants.
Ishizaki
Takashi
T
Accelrys KK, Nishishinbashi TS Building 11F, 3-3-1 Nishishinbashi, Minato-ku, Tokyo 105-0003, Japan.
Ohsumi
Chieko
C
Totsuka
Kazuhiko
K
Igarashi
Daisuke
D
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D002245:Carbon Dioxide / Q000008:administration & dosage; D019069:Cell Respiration / Q000235:genetics; D015870:Gene Expression; D018698:Glutamic Acid / Q000378:metabolism*; D006706:Homeostasis / Q000528:radiation effects; D008027:Light; D009566:Nitrates / Q000008:administration & dosage; D009584:Nitrogen / Q000378:metabolism; D035261:Plant Components, Aerial / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D036226:Seedlings / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects
D000596:Amino Acids; D029681:Arabidopsis Proteins; D009566:Nitrates; D012333:RNA, Messenger; D002245:Carbon Dioxide; D018698:Glutamic Acid; D002244:Carbon; D000594:Amino Acid Oxidoreductases; C042233:glutamate synthase (ferredoxin); D009584:Nitrogen