湖北海棠MhWRKY40b 在几种胁迫下的表达分析  

Expression Analysis of Malus hupehensis(Pamp) Rehd. MhWRKY40b Gene in Response to Several Stresses

作　　者：罗昌国[1,2] 渠慎春 张计育[3] 王三红 乔玉山 张仕杰 刘丹 章镇 

机构地区：[1]京农业大学园艺学院,南京210095 [2]贵州省农业科学院果树科学研究所,贵阳550006 [3]江苏省中国科学院植物研究所,南京210014

出　　处：《园艺学报》2013年第1期1-9,共9页Acta Horticulturae Sinica

基　　金：国家‘863’计划项目(2011AA100204);国家自然科学基金项目(31171935);现代园艺科学优势学科建设工程专项项目

摘　　要：为了分析 WRKY40 抗病抗逆作用,以湖北海棠[Malus hupehensis(Pamp)Rehd.]为试材,利用特异引物进行 RT-PCR 基因克隆,利用实时荧光定量 PCR 方法分析组织中基因的表达及低温、高盐、干旱、PEG6000 胁迫和外源 IAA、ABA 处理对基因表达的影响。结果表明,克隆到的片段长度为 1 240 bp,包含一个完整的开放阅读框,长 999 bp,编码 333 个氨基酸。该基因与拟南芥 AtWRKY40 同源,故命名为 MhWRKY40b,GenBank 登录号为 JX112913。基因表达分析表明,MhWRKY40b 在根、茎、叶等组织均有表达,其中在花萼、花辨、果实中表达量较高。该基因明显受低温、高盐、干旱胁迫诱导表达,最高相对表达量可达 20 倍以上;而受 PEG、苹果白粉病菌及外源 ABA 和 IAA 轻微诱导,最高相对表达量在10 倍以下。MhWRKY40b 除了在抗白粉病方面与 AtWRKY40 具有相似功能外,可能还在湖北海棠的抗寒、抗盐、抗旱过程中起到比较重要的作用。In order to reveal the role of WRKY40 gene in response to diseases and environmental stresses,Malus hupehensis（Pamp）Rehd. was used for gene cloning by reverse transcription PCR amplification by using specific primers. Quantitative real-time PCR was used in analyzing gene expression in tissues and in response to stresses,including cold,high salinity,drought,PEG6000 and treatments of exogenous ABA（abscisic acid）and IAA（indole-3-acetic acid）. A 1 240 bp fragment Arabidopsis AtWRKY40,and it is named as MhWRKY40b. Its GenBank accession is JX112913. Gene expression indicated that MhWRKY40b expressed in root,stem,leaf and other tissues,especially high in calyx,petal and fruit. It was clearly induced by cold,high salinity,and drought with more than 20 folds increased expression at the highest level,and slightly induced by PEG osmotic stress,Podosphaera leucotricha,exogenous ABA,and IAA,with less than 10 folds increase at the highest level. MhWRKY40b may have similar function to AtWRKY40 in powdery mildew resistance,it may also have a role in cold,salinity and drought tolerance.

关 键 词：湖北海棠 WRKY 转录因子 抗逆性 抗病性 ABA 

分 类 号：S661[农业科学—果树学]