棉花逆境应答GhWRKY2基因的结构与功能预测

发布时间：2019-05-10 20:20

【摘要】：WRKY是一类与植物抗逆相关的转录因子,通过激活下游一系列抗逆应答基因的表达而提高植株的综合抗性。本研究通过基因芯片从陆地棉(Gossypium hirsutum)中筛选出一个逆境诱导WRKY转录因子基因GhWRKY2,属于Ⅱ亚组。采用RT-PCR技术获得GhWRKY2基因cDNA全长,并对其进行序列分析及结构功能预测。根据GhWRKY2蛋白序列进行同源性搜索,得到与GhWRKY2蛋白序列相似度较高的其他物种的蛋白序列,使用MEGA7软件对GhWRKY2蛋白序列及其同源序列进行多序列比对分析并构建同源物种间系统进化树。利用在线工具Prot Param和Motif scan对GhWRKY2进行氨基酸理化性质分析;利用SMART在线工具进行蛋白序列分析;利用SWISS-MODEL在线工具分别对GhWRKY2基因结构和三级结构进行分析。结果表明,GhWRKY2核苷酸序列ORF为942 bp,编码313个氨基酸,预测蛋白分子量为34.22 k D,等电点为8.76。在拟南芥原生质体中,GhWRKY2定位于细胞核内,且具有转录激活活性。逆境应答GhWRKY2基因的克隆,为进一步从分子水平上验证其抗逆生物学功能以及揭示棉花非生物胁迫抗逆机制奠定基础,并为棉花抗逆育种提供新的候选基因。
[Abstract]:WRKY is a class of transcription factors related to plant stress resistance, which improves the comprehensive resistance of plants by activating the expression of a series of downstream stress response genes. In this study, a stress-induced WRKY transcription factor gene GhWRKY2, was screened from upland cotton (Gossypium hirsutum) by gene chip, which belongs to subgroup II. The full length of GhWRKY2 gene cDNA was obtained by RT-PCR, and its sequence analysis and structure and function prediction were carried out. According to the homology search of GhWRKY2 protein sequence, the protein sequences of other species with high similarity to GhWRKY2 protein sequences were obtained. The GhWRKY2 protein sequences and their homologous sequences were analyzed by MEGA7 software and the phylogenetic tree among homologous species was constructed. The physical and chemical properties of GhWRKY2 were analyzed by online tools Prot Param and Motif scan, the protein sequence was analyzed by SMART online tool, and the structure and tertiary structure of GhWRKY2 gene were analyzed by SWISS-MODEL online tool. The results showed that the ORF of GhWRKY2 nucleotides was 942 bp, encoding 313amino acids, the predicted protein molecular weight was 34.22kD, and the isoelectric point was 8.76. the results showed that the predicted protein molecular weight was 34.22 KD and the isoelectric point was 8.76%. In Arabidopsis protoplasts, GhWRKY2 is located in the nucleus and has transcriptional activation activity. The cloning of stress response GhWRKY2 gene laid a foundation for further verifying the biological function of stress resistance and revealing the mechanism of stress resistance in cotton at the molecular level, and provided a new candidate gene for cotton stress resistance breeding.
【作者单位】：新疆农垦科学院棉花研究所/农业部西北内陆区棉花生物学与遗传育种重点实验室/新疆兵团棉花遗传改良与高产栽培重点实验室;
【基金】：国家自然科学基金(No.31260360) 兵团青年科技创新资金(No.2011CB001)
【分类号】：Q943.2;S562

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