Lancien Muriel, Martin Melinda, Hsieh Ming-Hsiun, Leustek Tom, Goodman Howard, Coruzzi Gloria M
Department of Biology, New York University, 100 Washington Square East, 1009 Main Building, New York, NY 10003, USA.
Plant J. 2002 Feb;29(3):347-58. doi: 10.1046/j.1365-313x.2002.01218.x.
The physiological role of the NADH-dependent glutamine-2-oxoglutarate aminotransferase (NADH-GOGAT) enzyme was addressed in Arabidopsis using gene expression analysis and by the characterization of a knock-out T-DNA insertion mutant (glt1-T) in the single NADH-GOGAT GLT1 gene. The NADH-GOGAT GLT1 mRNA is expressed at higher levels in roots than in leaves. This expression pattern contrasts with GLU1, the major gene encoding Fd-GOGAT, which is most highly expressed in leaves and is involved in photorespiration. These distinct organ-specific expression patterns suggested a non-redundant physiological role for the NADH-GOGAT and Fd-GOGAT gene products. To test the in vivo function of NADH-GOGAT, we conducted molecular and physiological analysis of the glt1-T mutant, which is null for NADH-GOGAT, as judged by mRNA level and enzyme activity. Metabolic analysis showed that the glt1-T mutant has a specific defect in growth and glutamate biosynthesis when photorespiration was repressed by 1% CO2. Under these conditions, the glt1-T mutant displayed a 20% decrease in growth and a dramatic 70% reduction in glutamate levels. Herein, we discuss the significance of NADH-GOGAT in non-photorespiratory ammonium assimilation and in glutamate synthesis required for plant development.
利用基因表达分析以及对单个NADH依赖型谷氨酰胺-2-酮戊二酸氨基转移酶(NADH-GOGAT)基因中的敲除T-DNA插入突变体(glt1-T)进行表征,研究了拟南芥中NADH-GOGAT酶的生理作用。NADH-GOGAT GLT1 mRNA在根中的表达水平高于叶。这种表达模式与GLU1相反,GLU1是编码Fd-GOGAT的主要基因,在叶中表达最高且参与光呼吸。这些不同的器官特异性表达模式表明NADH-GOGAT和Fd-GOGAT基因产物具有非冗余的生理作用。为了测试NADH-GOGAT的体内功能,我们对glt1-T突变体进行了分子和生理分析,从mRNA水平和酶活性判断,该突变体的NADH-GOGAT缺失。代谢分析表明,当光呼吸被1%的CO2抑制时,glt1-T突变体在生长和谷氨酸生物合成方面存在特定缺陷。在这些条件下,glt1-T突变体的生长下降了20%,谷氨酸水平显著降低了70%。在此,我们讨论了NADH-GOGAT在非光呼吸铵同化以及植物发育所需的谷氨酸合成中的重要性。