铁皮石斛DoCDPK6基因及启动子的克隆与表达分析

发布时间：2018-05-05 04:34

本文选题：铁皮石斛 + 铁皮石斛钙依赖蛋白激酶基因(DoCDPK)　；参考：《农业生物技术学报》2017年04期

【摘要】：钙依赖蛋白激酶(calcium-dependent protein kinase,CDPK)是一类能响应低温等非生物胁迫的蛋白激酶,目前已从拟南芥(Arabidopsis thaliana)、水稻(Oryza sativa)和杨树(Populus trichocarpa)等植物中分离出多个CDPK基因。为了研究铁皮石斛(Dendrobium officinale)钙依赖蛋白激酶的结构和功能,探寻其在逆境胁迫下的作用,从铁皮石斛中分离出1个DoCDPK6基因(Gen Bank No.KY682702)及其启动子,通过生物信息学分析序列特征、结构及进化关系,结果表明,DoCDPK6基因的cDNA全长1 976 bp,开放阅读框1 725 bp,编码574个氨基酸,分子式为C2845H4500N766U858S24,分子量为63.9 k D;具有CDPK典型的保守结构:可变区、蛋白激酶区、连接区和钙调控区;进化分析表明,DoCDPK6与同科植物蝴蝶兰(Phalaenopsis amabilis)亲缘关系最近,符合植物学分类地位;在DoCDPK6基因1 500 bp的上游序列含有核心启动子元件TAAT-box和CAAT-box,并含有光、温、水杨酸、脱落酸、生长素和赤霉素等逆境信号相关的响应元件;通过q RT-PCR发现,铁皮石斛DoCDPK6主要在叶片中表达,其次是茎和根;在4℃低温、脱落酸(abscisic acid,ABA)和Na Cl胁迫处理后,DoCDPK6基因表达量均有不同程度的上调,因此可以推测,DoCDPK6基因在低温等非生物胁迫中起到调控作用,该研究结果可为DoCDPK6基因及其启动子在非生物胁迫中的调控机制和信号传导途经提供信息。
[Abstract]:Calcium-dependent protein kinase (CDPKK) is a kind of protein kinase that can respond to abiotic stress such as low temperature. At present, many CDPK genes have been isolated from Arabidopsis thaliana, Oryza sativa and Populus roseus. In order to study the structure and function of calcium-dependent protein kinase of Dendrobium officinaleum, and to explore its role in stress, a DoCDPK6 gene, Genin Bank No. KY682702) and its promoter were isolated from Dendrobium officinale. and the sequence characteristics were analyzed by bioinformatics. The results of structure and evolution showed that the cDNA length of DoddPK6 gene was 1 976 BP, open reading frame 1 725 BP, encoding 574 amino acids, the molecular formula was C2845H4500N766U858S24, and its molecular weight was 63.9 KD, which had the typical conserved structure of CDPK: variable region, protein kinase region. Phylogenetic analysis showed that DoCDPK6 was closely related to Phalaenopsis amabilis (Phalaenopsis amabilis) of the family Phalaenopsis. The upstream sequence of DoCDPK6 gene (1 500bp) contained TAAT-box and CAAT-box, which contained light and temperature. Salicylic acid, abscisic acid, auxin and gibberellin, and so on. By Q RT-PCR, it was found that DoCDPK6 was mainly expressed in leaves, followed by stems and roots. The expression of DoCDPK6 gene was up-regulated by abscisic acidad (Aba) and NaCl stress in different degrees, so it can be inferred that DoCDPK6 gene plays a regulatory role in abiotic stress such as hypothermia. The results may provide information for the regulation mechanism and signal transduction pathway of DoCDPK6 gene and its promoter in abiotic stress.
【作者单位】：浙江农林大学亚热带森林培育国家重点实验室培育基地;
【基金】：国家自然科学基金(No.81603228)
【分类号】：S567.239

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