陆地棉盐胁迫应答基因GhWRKY41的克隆与分析  

Cloning and analysis of a salt stress responsive gene GhWRKY41 in upland cotton(Gossypium hirsutum L.)

作　　者：苏莹[1] 甄军波[1] 张曦[1] 王玉美[2] 李乐[1] 华金平[1] SU Ying ZHEN Jun-bo ZHANG Xi WANG Yu-mei LI Le HUA Jin-ping(College of Agronomy/Key Laboratory of Crop Heterosis and Utitization of Ministry of Education/ ng Key Laboratory of Crop Genetic Improvement , China Agricultural University, Beijing 100193, China Institute of Cash Crop, Hubei Academy of Agricultural Sciences, Wuhan 430064, China)

机构地区：[1]中国农业大学农学院/杂种优势研究与利用教育部重点实验室/作物遗传改良北京市重点实验室,北京100193 [2]湖北省农业科学院经济作物研究所,武汉430064

出　　处：《中国农业大学学报》2016年第12期1-11,共11页Journal of China Agricultural University

基　　金：国家转基因重大专项(2014ZX08005-004;2014ZX08005-004B);国家高技术研究发展计划(863计划)重大项目(2006AA10A108)

摘　　要：为研究转录因子GhWRKY41在陆地棉盐胁迫应答过程中的作用,基于差减文库分析结果,利用RT-PCR和RACE技术,克隆了GhWRKY41基因(GenBank登录号为HM002635)。该基因cDNA长度为1 630bp,含有ORF(Open reading frame)为1 068bp,编码355个氨基酸的多肽,包含2个内含子。通过瞬时表达分析亚细胞定位,结果表明,转录因子GhWRKY41定位于细胞核,符合转录因子特性。转基因株系发芽试验结果表明,过量表达GhWRKY41基因,可显著提高转基因棉花在干旱、盐和低温胁迫下的发芽率;利用Real-time PCR技术,证明在盐和干旱胁迫条件下,转基因株系中GhWRKY41基因的表达量显著上升。GhWRKY41基因在根、茎和叶片中表达存在差异,根系中胁迫6h上调达到最高,茎中则胁迫48h达到最高,而叶片中仅6和24h上调表达。进一步比较转基因棉花与野生型棉花的纤维品质性状,结果表明GhWRKY41的过表达可以提高转基因棉花的衣分。因此,GhWRKY41参与了棉花响应盐和干旱胁迫应答过程,且过表达可提高转基因棉花耐盐性和耐旱性。In order to study the role of transcription factor GhWRKY41 in salt stress responsive progress of upland cotton（Gossypium hirsutum L.）,aGhWRKY41 gene（Accession No.HM002635）was isolated from upland cotton by using RT-PCR and RACE methods based on SSH library analyze.The full-length of cDNA sequence is 1 630 bp,and has a 1 068 bp ORF with two introns,coding 355 bp amino acids.Transient expression assay showed that GhWRKY41 was subcellular localized in nucleus.Germination rate analysis showed that overexpression of GhWRKY41 gene could significantly increase the germination rate of transgenic cotton lines under abiotic treatments of drought,salt and cold stresses.The results of Real-time PCR proved that the GhWRKY41 gene could be induced by salt and drought treatments.And there were differences in the expression profile of GhWRKY41 gene in root,stem and leaf tissue.The expression level of GhWRKY41 gene after salt treatment was highest at 6hin root,was highest at 48 hafter treatment in stem,and was up-regulated at 6and 24 hin leaves.The fiber quality traits of transgenic lines to receptor line were compared.And it was discovered that overexpressed GhWRKY41 could improve the lint percentage of transgenic cotton.In conclusion,GhWRKY41 was involved in plant response to salt and drought stress in cotton,and its overexpression in plant improved tolerance to salt and drought stresses.

关 键 词：陆地棉 GhWRKY41 盐胁迫 表达分析 发芽率 

分 类 号：S661.1[农业科学—果树学]