陆地棉转录因子基因GhC2H2的克隆与功能分析  

Cloning and Functional Analysis of a Transcription Factor Gene, GhC2H2, in Upland Cotton(Gossypium hirsutum L.)

作　　者：苏莹[1] 甄军波[1,2] 张曦[1] 王玉美[3] 华金平[1] 

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]河北省农林科学院棉花研究所/农业部黄淮海半干旱区棉花生物学与遗传育种重点实验室,河北石家庄050051 [3]湖北省农业科学院经济作物研究所,湖北武汉430064

出　　处：《棉花学报》2016年第6期555-564,共10页Cotton Science

基　　金：国家高技术研究发展计划(863计划)重大项目(2006AA10A108);国家科技重大专项--转基因生物新品种培育(2014ZX08005-004B)

摘　　要：C2H2型锌指蛋白在植物生长发育、非生物胁迫响应过程中起关键作用。基于实验室前期工作,分析盐胁迫下陆地棉根系抑制性差减杂交文库,筛选得到1个盐胁迫应答基因GhC2H2。结合RACE(Rapid amplification of cDNA ends)和RT-PCR(Reverse transcription-polymerase chain reaction)技术克隆该基因全长,通过瞬时表达分析其细胞定位,采用Real time-PCR分析其在盐胁迫下的陆地棉根、茎、叶中的表达模式,并测定了转入该基因的株系在正常土壤和盐碱地的铃重、衣分和纤维品质指标。该基因GenBank登录号为HM002632,编码区全长819 bp,编码272 aa;其编码蛋白包含2个C2H2锌指结构域,定位于细胞核;该基因在不同组织中均受盐胁迫诱导上调表达,且在胁迫诱导初期上调量最大,推测该基因参与早期盐胁迫应答。对转基因后代株系性状分析表明,该基因的过表达可以提高转基因棉花衣分、改良纤维品质,推测GhC2H2既参与盐胁迫应答过程,也参与纤维发育调控。C2H2 zinc finger proteins play key roles in plant growth, development and abiotic stress-response processes. Based on a previous study of the suppression subtractive hybridization libraries of upland cotton(G. hirsutum L.) roots under salt stress, a salt stress-response gene, Gh C2H2, was selected. By combining rapid amplification of c DNA ends and reverse transcription-polymerase chain reaction methods, the full length of Gh C2H2(Gen Bank Accession No. HM002632) was isolated. It had an open reading frame region of 819 bp that encoded 272 aa. The Gh C2H2 protein contains two C2H2 zinc finger domains. A transient expression analysis indicated that the transcription factor was localized in the nucleus. An expression profile analysis showed that the gene was up-regulated in different tissues(root, stem and leaf) of upland cotton under salt stress, and we hypothesized that Gh C2H2 may be involved in the early stage of the salt-stress response. A phenotypic trait analysis of transgenic cotton lines in two environments(normal and saline-alkali soil) showed that overexpressed GhC2H2 could improve the lint percentage as well as the fiber quality. Thus, we hypothesized that GhC2H2 participates in both the salt-stress responsive process and fiber development.

关 键 词：陆地棉 GhC2H2位 基因克隆 盐胁迫应答 基因功能 

分 类 号：S562[农业科学—作物学]