Instituto de Bioingeniería, Universidad Miguel Hernández, Campus de Elche, 03202, Elche, Spain.
Sci Rep. 2017 Jul 21;7(1):6164. doi: 10.1038/s41598-017-06335-4.
Biomass production requires the coordination between growth and metabolism. In a large-scale screen for mutants affected in leaf morphology, we isolated the orbiculata1 (orb1) mutants, which exhibit a pale green phenotype and reduced growth. The combination of map-based cloning and next-generation sequencing allowed us to establish that ORB1 encodes the GLUTAMATE SYNTHASE 1 (GLU1) enzyme, also known as FERREDOXIN-DEPENDENT GLUTAMINE OXOGLUTARATE AMINOTRANSFERASE 1 (Fd-GOGAT1). We performed an RNA-seq analysis to identify global gene expression changes in the orb1-3 mutant. We found altered expression levels of genes encoding enzymes involved in nitrogen assimilation and amino acid biosynthesis, such as glutamine synthetases, asparagine synthetases and glutamate dehydrogenases, showing that the expression of these genes depends on the levels of glutamine and/or glutamate. In addition, we observed a concerted upregulation of genes encoding subunits of the cytosolic ribosome. A gene ontology (GO) analysis of the differentially expressed genes between Ler and orb1-3 showed that the most enriched GO terms were 'translation', 'cytosolic ribosome' and 'structural constituent of ribosome'. The upregulation of ribosome-related functions might reflect an attempt to keep protein synthesis at optimal levels even when the pool of glutamate is reduced.
生物量的生产需要生长和代谢之间的协调。在一个大规模的叶片形态突变体筛选中,我们分离到 orbiculata1(orb1)突变体,其表现出浅绿色表型和生长减缓。通过图谱定位克隆和新一代测序的结合,我们确定 ORB1 编码 GLUTAMATE SYNTHASE 1(GLU1)酶,也称为 FERREDOXIN-DEPENDENT GLUTAMINE OXOGLUTARATE AMINOTRANSFERASE 1(Fd-GOGAT1)。我们进行了 RNA-seq 分析,以鉴定 orb1-3 突变体中的全局基因表达变化。我们发现参与氮同化和氨基酸生物合成的酶的基因表达水平发生了改变,如谷氨酰胺合成酶、天冬酰胺合成酶和谷氨酸脱氢酶,表明这些基因的表达依赖于谷氨酰胺和/或谷氨酸的水平。此外,我们观察到胞质核糖体亚基的基因表达协同上调。Ler 和 orb1-3 之间差异表达基因的基因本体(GO)分析表明,最富集的 GO 术语是“翻译”、“胞质核糖体”和“核糖体结构成分”。核糖体相关功能的上调可能反映了即使谷氨酸池减少,也要保持蛋白质合成处于最佳水平的尝试。