刚毛柽柳ThγGCS基因克隆及表达分析

发布时间：2018-08-28 10:10

【摘要】：γ-谷氨酰半胱氨酸合成酶(γGCS)是植物细胞内谷胱甘肽(GSH)生物合成的限速酶,在植物许多的生理过程中具有重要作用。从刚毛柽柳转录组数据库中分析并获得1条刚毛柽柳γGCS基因全长c DNA序列,命名为ThγGCS。该基因全长2 121 bp,开放阅读框为1 536 bp,编码511个氨基酸。生物信息学分析结果表明,ThγGCS推导的氨基酸序列分别与苜蓿、大豆以及橡胶树等具有很高的同源性;进化分析结果表明ThγGCS属双子叶植物γGCS亚类,同橡胶树,芥菜等γGCS分为一组,与单子叶植物的亲缘关系较远。该蛋白质无跨膜结构域,但具有1个保守的GCS2结构域,并被定位于叶绿体中,说明ThγGCS蛋白质较稳定,并属于谷氨酰半胱氨酸连接酶家族。为了研究该基因在不同非生物胁迫下的应答情况,利用实时定量RT-PCR技术,分析ThγGCS在NaCl和PEG胁迫处理后不同时间、不同组织中基因的表达。结果表明,ThγGCS受NaCl、PEG诱导上调表达,推测其可能在刚毛柽柳抗旱耐盐过程中发挥重要作用。
[Abstract]:纬-glutamyl cysteine synthase (纬 GCS) is a rate-limiting enzyme for glutathione (GSH) biosynthesis in plant cells and plays an important role in many physiological processes in plants. A full-length c DNA sequence of Tamarix chinensis 纬 GCS gene was obtained from Tamarix chinensis transcriptome database and named Th 纬 GCS.. The full-length 2 121 bp, open reading frame encodes 511 amino acids for 1 536 bp,. The results of bioinformatics analysis showed that the deduced amino acid sequences of Th 纬 GCS were highly homologous to alfalfa, soybean and Hevea, respectively, and the evolutionary analysis showed that Th 纬 GCS was a dicotyledonous plant 纬 GCS subclass, which was divided into the same group as Hevea, mustard and other 纬 GCS. The phylogenetic relationship with monocotyledonous plants is far away. The protein has no transmembrane domain, but has a conserved GCS2 domain and is located in chloroplast, indicating that the Th 纬 GCS protein is stable and belongs to the glutamyl cysteine ligase family. In order to study the response of the gene to different abiotic stresses, the expression of Th 纬 GCS in different tissues at different time after NaCl and PEG stress was analyzed by real-time quantitative RT-PCR. The results showed that the expression of Th 纬 GCS was up-regulated by NaCl,PEG, which suggested that Th 纬 GCS might play an important role in the process of drought and salt tolerance of Tamarix chinensis.
【作者单位】：东北林业大学林木遗传育种国家重点实验室;
【基金】：中央高校基本科研业务费专项资金项目(2572014CA12) 东北林业大学大学生创新训练项目(2015102251-81)共同资助
【分类号】：S793.5;Q943.2

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