漾濞大泡核桃WRKY转录因子基因JsWRKY1的克隆及表达特性分析  

Cloning and Expression Analysis of a WRKY Gene JsWRKY1 from Juglans sigillata

作　　者：张南南[1] 陈朝银[1] 季博[1] 何华[1] 韩青[1] 葛锋[1] 刘迪秋[1] 

机构地区：[1]昆明理工大学生命科学与技术学院,昆明650500

出　　处：《植物生理学报》2014年第7期960-966,共7页Plant Physiology Journal

基　　金：科技部支撑计划项目(2011BAD46B00)

摘　　要：WRKY转录因子普遍存在于植物体内,在植物的抗病防御反应中起重要作用。本实验基于漾濞大泡核桃(Juglans sigillata)中编码WRKY转录因子的EST序列设计引物,采用快速扩增cDNA末端技术,克隆得到一个新的WRKY基因的全长cDNA序列,命名为JsWRKY1(KJ170895)。JsWRKY1的cDNA全长为1 012 bp,含有564 bp的开放阅读框,154 bp 5'-非翻译区以及294 bp的3?-非翻译区,编码具有187个氨基酸的蛋白质。JsWRKY1编码的氨基酸序列与已知植物WRKY家族成员间的同源性和聚类分析表明JsWRKY1与来源于可可树(Theobroma cacao)和大豆(Glycine max)中的WRKY相似性较高,属于IIc类WRKY。qRT-PCR分析结果显示,信号分子水杨酸、茉莉酸、H2O2和乙烯处理可以不同程度地诱导漾濞大泡核桃叶片中JsWRKY1的表达。此外,接种胶孢炭疽菌后JsWRKY1的表达量迅速上升,在接种后4 h时达到最高水平,之后表达量逐渐下降,暗示JsWRKY1参与漾濞大泡核桃抗胶孢炭疽菌的防卫反应。The WRKY transcription factors are ubiquitous in plants and play important roles in the defense responses. A WRKY EST was isolated from Juglans sigillata, and the full-length cDNA of the EST was cloned with the method of rapid amplification of cDNA ends and named as Js WRKY1 （KJ170895）. Js WRKY1 was 1 012 bp in length and contained an intact open reading frame （ORF） of 564 bp, a 154 bp 5＇-untranslated region （UTR）, and a 294 bp 3＇-UTR. The predicted protein of Js WRKY1 with 187 amino acid residues shared high similarity with the WRKYs from Theobroma cacao and Glycine max. Moreover, the JsWRKY1 fell into the group IIc of WRKYs through phylogenetic analysis, qRT-PCR analysis showed that the expression ofJs WRKY1 was induced by salicylic acid, jasmonic acid, H2O2, and ethylene. Morever, Js VRKY1 was quickly induced after inoculation with Colletorichum gloeosporidies, and the highest transcription level was achieved at 4 h post inoculation, then its expression declined gradually. All the results of present study suggested that Js WRKY1 may involve in defense response ofJ. sigillata against the C. gloeosporioides.

关 键 词：漾濞大泡核桃 WRKY转录因子 基因克隆 胶孢炭疽菌 防卫反应 

分 类 号：S664.1[农业科学—果树学]