Glutamine synthetase and glutamate dehydrogenase in triticale seeds: molecular cloning and genes expression
Glutamine synthetase and glutamate dehydrogenase in triticale seeds: molecular cloning and genes expression
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DOI：
10.1007/s11738-012-1085-9
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发表时间：
2012-11-01
影响因子：
2.6
通讯作者：
Bielawski, Wieslaw
中科院分区：
文献类型：
--
作者：
Grabowska, Agnieszka;Kwinta, Joanna;Bielawski, Wieslaw
在高等植物中，谷氨酰胺合成酶（GS; EC 6.3.1.2）和谷氨酸脱氢酶（GDH; EC 1.4.1.2）是氮代谢中的主要酶。在这项研究中，我们克隆了GS和GDH基因，并分析了它们的表达水平和活性的变化在发育和萌发x小黑麦（cv. Witon）内核。在开花后3、5、7、9、13、15、20、25、30、35、40和45天收集发育中的籽粒样品。在吸胀8、16、24、48和72 h后收集发芽的籽粒样品。有两种GS亚型定位于不同的隔室：细胞质（GS 1）和叶绿体（GS 2）。利用RT-PCR技术获得了5个编码小黑麦GS蛋白的cDNA。我们克隆了4个编码GS1的基因，命名为TsGS 1 -1、TsGS 1 -2、TsGS 1 -3和TsGS 1 -4，并将唯一编码GS 2的基因命名为TsGS 2 -1。研究了小黑麦种子发育和萌发过程中GDH、GS1和GS 2基因的酶活性和表达谱的变化。根据我们的研究结果，有可能合作GDH和GS 1之间的谷氨酰胺和谷氨酸盐的合成在种子形成的早期阶段，并在盾片的内核长达24小时的吸胀。
In higher plants, glutamine synthetase (GS; EC 6.3.1.2) and glutamate dehydrogenase (GDH; EC 1.4.1.2) are the predominant enzymes in nitrogen metabolism. In this study, we cloned both the GS and GDH genes and analyzed their expression levels and variations in their activity in developing and germinating x Triticosecale (cv. Witon) kernels. The developing kernel samples were collected 3, 5, 7, 9, 13, 15, 20, 25, 30, 35, 40 and 45 days after flowering (DAF). The germinating kernel samples were collected after 8, 16, 24, 48 and 72 h of imbibition. There are two GS isoforms that are localized to different compartments: the cytosol (GS1) and the chloroplast (GS2). Five cDNAs encoding GS proteins in triticale plants were obtained using RT-PCR. We cloned the four genes encoding GS1, which we designated TsGS1-1, TsGS1-2, TsGS1-3 and TsGS1-4 and the only gene encoding GS2, which was designated TsGS2-1. We studied the changes in the enzymatic activity and the expression profiles of the GDH, GS1 and GS2 genes in both the developing and germinating seeds of triticale. Based on our results, there is likely cooperation between GDH and GS1 in the synthesis of glutamine and glutamate during the early stages of seed formation and in the scutella of kernels for up to 24 h of imbibition.