橡胶树转录因子HbWRKY9的克隆与特性分析  

Cloning and Characteristics of HbWRKY9 Transcription Factor in the Rubber Tree

作　　者：赵武帅 翟琪麟[2] 安泽伟[2] 方家林[2] 黄华孙[2] 

机构地区：[1]海南大学农学院,海口570228 [2]国家橡胶树育种中心,中国热带农业科学院橡胶研究所,儋州571737

出　　处：《基因组学与应用生物学》2015年第3期599-606,共8页Genomics and Applied Biology

基　　金：国家天然橡胶产业技术体系(No.CARS-34);国家自然科学基金(No.30960319;3127196)共同资助

摘　　要：通过RACE技术克隆得到一个新的橡胶树WRKY,命名为Hb WRKY9,Gen Bank登录号为JQ914653,基因全长为1 646 bp,ORF为954 bp,编码317个氨基酸。Hb WRKY9蛋白含有1个WRKY结构域和1个C2H2型锌指结构,与蓖麻(Ricinus communis)、麻疯树(Jatropha carcas)、毛果杨(Populus trichocarpa)和酿酒葡萄(Vitis vinifera)的WRKY成员一致性分别为80%、79%、70%和63%。对其启动子序列的分析表明,启动子核心区域位于翻译起始密码子上游74~123 bp处,启动子序列中包含TATA-box、CAAT-box、5’UTR Py-rich stretch、WRKY转录因子结合位点W-box、MYB结合位点MBS,以及与光响应相关的调控元件G-box和Box4,分生组织特异表达调控元件CCGTCC-box,与缺氧诱导相关的ARE元件,与高温胁迫相关的调控元件HSE,与Me JA和ABA等激素响应相关的CGTCA-motif和ABRE等顺式调控元件。实时荧光定量PCR分析表明,Hb WRKY9基因能响应低温胁迫,呈上调表达。In this study, Hb WRKY9 was cloned from the rubber tree by RACE technology. Hb WRKY9 was 1 646 bp in length and contained a 954 bp open reading frame encoding 317 amino acids. Hb WRKY9 protein contained one WRKY domain and C2H2 zinc finger domain. Alignment analysis showed that the amino acid sequence of Hb WRKY9 is 80%, 79%, 70% and 63% similar with that of Ricinus communis, Jatropha carcas, Populus trichocarpa and Vitis vinifera, respectively. A promoter core region was found between 74 bp to 123 bp in the upstream of initiation codon ATG, and some elements and motifs were found in the promoter sequence of Hb WRKY9, such as CAAT box, 5'UTR Py-rich stretch, W-box(a WRKY transcription factor binding site), MBS(a MYB binding site),light responsive elements(G-box and Box4), meristematic tissue specific expression element(CCGTCC-box),regulatory element essential for the anaerobic induction(ARE), high temperature responsive element(HSE),Me JA and ABA responsive element(CGTCA-motif and ABRE). Real time quantitative PCR analysis showed that Hb WRKY9 was induced by low temperature and the expression was up-regulated.

关 键 词：橡胶树 转录因子 HbWRKY9 非生物胁迫 低温 

分 类 号：S794.1[农业科学—林木遗传育种] Q943.2[农业科学—林学]