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棉花学报 ›› 2016, Vol. 28 ›› Issue (6): 555-564.doi: 10.11963/issn.1002-7807.201606005

• 研究与进展 • 上一篇    下一篇

陆地棉转录因子基因GhC2H2的克隆与功能分析

苏莹1(),甄军波1,,张曦1,王玉美2,*(),华金平1   

  1. 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,湖北 武汉430064
  • 收稿日期:2016-02-17 出版日期:2016-11-15 发布日期:2016-11-15
  • 通讯作者: 甄军波,王玉美 E-mail:suying0425@163.com;yumeiwang002@126.com
  • 作者简介:苏莹(1986―),女,博士研究生,suying0425@163.com
  • 基金资助:
    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)

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

Ying Su1(),Junbo Zhen1,,Xi Zhang1,Yumei Wang2,*(),Jinping Hua1   

  1. 1. College of Agriculture, China Agricultural University/Key Laboratory of Crop Heterosis and Utilization of Ministry of Education/Beijing Key Laboratory of Crop Genetic Improvement, Beijing 100193, China
    2. Institute of Cash Crop, Hubei Academy of Agricultural Sciences, Wuhan, Hubei 430064, China
  • Received:2016-02-17 Online:2016-11-15 Published:2016-11-15
  • Contact: Junbo Zhen,Yumei Wang E-mail:suying0425@163.com;yumeiwang002@126.com

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。

关键词: 陆地棉; GhC2H2; 基因克隆; 盐胁迫应答; 基因功能

Abstract:

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, GhC2H2, was selected. By combining rapid amplification of cDNA ends and reverse transcription-polymerase chain reaction methods, the full length of GhC2H2 (GenBank Accession No. HM002632) was isolated. It had an open reading frame region of 819 bp that encoded 272 aa. The GhC2H2 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 GhC2H2 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.

Key words: upland cotton; GhC2H2; gene cloning; salt stress response; gene function