陆地棉基因GhWRKY40-1的克隆及表达分析.pdf

棉 花 学报 Cotton Science 2017, 29(2): 166―176 陆地棉基因 40-1 的克隆及表达分析 司爱君 田琴 陈红 余渝 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/ 疆兵团棉花遗传改良与 高产栽培重点实验室, 疆石河子832000 冤 摘要院揖目的铱WRKY 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遥 本研究旨在 研究WRKY 转录因子在陆地棉中的功能遥揖方法铱通过基因芯片筛选鉴定出1 个逆境诱导的陆地棉WRKY 转 录因子基因袁由于此基因编码蛋 含有1 个典型的WRKY 结构域袁且与拟南芥 40 具有较高相似性袁 故命名为 40-1遥 采用电子克隆及反转录- 聚合酶链反应技术获得 40-1 基因cDNA 全长袁 并对其进行序列分析及结构与功能预测遥 揖结果铱 40-1 的开放阅读框为981 bp 袁编码326 个氨基酸袁 3 预测蛋 相对分子质量为35.78 10 袁等电点为8.39遥GhWRKY40-1 在拟南芥原生质体中瞬时表达定位于细胞 核袁 且具有转录激活活性遥 RT-PCR 结果表明袁 40-1 受甘露醇诱导上调表达遥 揖结论铱 推测 40-1 可能参与干旱胁迫应答反应袁并且在植物逆境胁迫中发挥重要作用遥 上述分析为进一步从分子水 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 定了基础遥 关键词院棉花曰WRKY 曰转录因子曰基因克隆曰序列分析 中图分类号院S562.035 文献标志码院A 文章编号院1002-7807渊2017 冤02-0166-11 10.11963/issn.1002-7807.20 1702006 Si Aij un, Tian Qin, Chen Hong, Yu Yu ( 832000, ) [Objective] WRKY transcription factors enhance plant resistance by activating the expression of a series of stress-re- sponsive downstream genes. The purpose of our research was to analyze functions of WRKY transcription factors in upland cot- ton. [Method] A stress-induced upland-cotton WRKY transcription factor gene was screened by cDNA microarray analysis. The gene, which was found to contain a typical WRKY domain and to share a highly similar amino acid sequence with 40, was accordingly named 40-1. The full-length cDNA of GhWRKY40-1 was obtained by electronic cloning and reverse transcription-polymerase chain reaction (RT-PCR) technology