番茄WRKY41基因的克隆、表达分析与转基因植株的获得  

Cloning and Expression Analysis of a WRKY41 Gene in Tomato and Its Transfer into a Tomato Cultivar

作　　者：魏娟娟[1] 杨伟[1] 潘宇[1] 张兴国[1] 李金华[1] 

机构地区：[1]西南大学园艺园林学院/南方山地园艺学重点实验室,重庆400715

出　　处：《西南大学学报（自然科学版）》2017年第1期46-54,共9页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(31301779);西南大学博士启动金项目(SWU113023);中央高校基本业务费专项基金项目(XDJK2014B045)

摘　　要：基于前期基因芯片结果,RT-PCR获得栽培品种番茄M82和近缘野生种潘那利的一个WRKY基因的全长cDNA序列,分别命名为SlWRKY41和SpWRKY41.序列分析表明,番茄WRKY41基因长为1011bp,编码336个氨基酸.该氨基酸序列含有5'-N端WRKYGQK核心结构域和CX_7CX_(23)HX_1C锌指结构,具有WRKY家族的典型结构特征.同源分析表明,该氨基酸序列与多种植物的WRKY类蛋白具有较高的同源性,并且在普通栽培番茄品种M82和野生种潘那利中,只有5个氨基酸位点的差异.进化树分析表明,WRKY41在番茄中属于第Ⅲ类WRKY蛋白,这类蛋白为植物所特有.表达分析结果表明,WRKY41在普通栽培番茄品种M82和野生种潘那利中,不仅在不同组织器官中的表达存在差异,而且在逆境(干旱和氧化逆境)和一些调节因子(SA、GA、乙烯)的处理下也有不同的表达模式.其在野生种潘那利中能够迅速响应相关调节因子(SA、GA、乙烯),推测WRKY41在番茄抗逆响应过程中具有很重要的作用,Sp WRKY41可能是一个较好的改良普通栽培种抗逆性的候选基因.通过构建超量表达载体,成功地将SpWRKY41转化到番茄M82中,以期深入研究该基因的功能和提高番茄的抗逆性.Based on the results of microarray transcriptional analysis,a full-length c DNA sequence of the WRKY gene from M82,a cultivar of Solanum lycopersicum,and from the wild species S.pennellii LA0716 by RT-PCR,which were named as Sl WRKY41 and Sp WRKY41,respectively.Sequence analysis showed that Sl WRKY41 and SpWRKY41 gene contained a 1 011-bp open reading frame（ORF） encoding 336 amino acids.As the typical structure features of the WRKY family,a 5＇-N WRKYGQK domain and a central zinc finger region CX_7CX_（23）HX_1 C were identified in the amino acid sequence of WRKY41 protein.The deduced Sl WRKY41 and Sp WRKY41 protein was found to be highly homologous to the WRKY proteins from many other plants,and these two putative amino acids shared 97.62% similarity,differing only in five residues.Phylogenetic tree analysis indicated that WRKY41 fell into Group Ⅲ,which is unique to plants.The real-time PCR results suggested that the WRKY41 gene showed differential expression patterns in M82 and LA0716 not only in different tissues and organs but also in various stress or regulator treatments.Interestingly,Sp WRKY41 was responded very quickly by SA,GA and Eth.It is conjectured that Sp WRKY41 may play a crucial role in resistance to abiotic stress and is a good candidate gene for improving stressresistance of cultivated tomato.Furthermore,an overexpression vector was constructed,which then successfully transferred Sp WRKY41 to tomato M82 to enhance its abiotic tolerance.

关 键 词：番茄 野生种潘那利 WRKY41 表达模式 抗旱 

分 类 号：Q785[生物学—分子生物学] S641.2[农业科学—蔬菜学]