湖北海棠MhWRKY42基因克隆及表达分析  

Cloning and expression of MhWRKY42 gene from Malus hupehensis

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

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

出　　处：《南京农业大学学报》2013年第3期29-35,共7页Journal of Nanjing Agricultural University

基　　金：国家863计划项目(2011AA100204);现代园艺科学优势学科建设工程专项资金

摘　　要：从湖北海棠上克隆到1个WRKY基因,开放阅读框(ORF)长度为1 818个核苷酸,编码606个氨基酸。该基因氨基酸序列与拟南芥AtWRKY42的一致性为39.38%,具有WRKY基因家族典型保守结构域,即N端为WRKYGQK,C端为Cx4-5Cx22-23HxH;蛋白三级结构和拟南芥AtWRKY42相似,具有4个β折叠;系统进化树上与AtWRKY42聚在同一分支,属于WRKY基因家族GroupⅡb成员,命名为MhWRKY42(GenBank登录号:JX112905)。实时荧光定量RT-PCR分析表明,MhWRKY42基因在各个器官中都有表达,其中在果实中表达量最高,根和总花次之;在花中,以花萼表达量最高,子房和雄蕊次之;其受盐、低温、苹果白粉病诱导表达;外源IAA处理诱导其下调表达;而MhWRKY42基因表达受ABA、水分胁迫的影响小。A WRKY transcription factor was isolated from Malus hupehensis. Its ORF is 1 818 nucleotides and codes 606 amino acids. It shares 39.38% identity in amino acid sequence with Arabidopsis thaliana AtWRKY42. Its protein has two typical conserve domains of WRKY transcription factors superfamily with WRKYGQK at N terminal and Cx4-5 Cx22-23 HxH at C terminal, the tertiary structure is very similar to AtWRKY42 with 4 β sheet and the evolutionary tree indicates that this gene and AtWRKY42 are in the same branch. It is a member of Group Ⅱ b of WRKY transcription factors superfamily. It was designated as MhWRKY42 （ GenBank accession： JX112905 ）. Quantitative real-time PCR analysis（qRT-PCR） showed that MhWRKY42 gene expressed in all tissues, that the highest level was in fruit among organisms following root and flower, and that in flower, the highest was in sepal following ovary and anther. Among biotic stresses and abiotic stresses,MhWRKY42 gene was upregulated in salinity,cold and Podosphaera leucotricha stress. It was downregulated by exogenous IAA treatment, but just a little affected by exogenous ABA treatment and water stress.

关 键 词：湖北海棠 WRKY转录因子 基因克隆 表达 胁迫 

分 类 号：S661[农业科学—果树学]