陆地棉转录因子基因GhC2H2的克隆与功能分析
发布时间：2018-12-27 14:23
【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。
[Abstract]:C2H2 zinc finger protein plays a key role in plant growth and development, abiotic stress response. Based on the previous work in laboratory, the subtractive hybridization library of root inhibition of Upland cotton under salt stress was analyzed, and a salt stress response gene GhC2H2. was screened out. The full length of the gene was cloned by RACE (Rapid amplification of cDNA ends) and RT-PCR (Reverse transcription-polymerase chain reaction), and its cellular localization was analyzed by transient expression analysis. The expression patterns of the gene in root, stem and leaf of upland cotton under salt stress were analyzed by Real time-PCR. The boll weight, lint percentage and fiber quality index of the transgenic lines in normal soil and saline soil were determined. The GenBank accession number of the gene was 819 bp, encoding 272 aa;. The encoded protein contained two C2H2 zinc-finger domains and was located in the nucleus. The expression of the gene was up-regulated in different tissues induced by salt stress, and the up-regulation was the highest in the early stage of stress induction. It was speculated that the gene was involved in the early salt stress response. The analysis of the characters of transgenic progenies showed that the overexpression of the gene could improve lint score and fiber quality of transgenic cotton. It was inferred that GhC2H2 was involved in both salt stress response and fiber development regulation.
【作者单位】： 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;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;湖北省农业科学院经济作物研究所;
【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)
【分类号】：S562
，
本文编号：2393206
[Abstract]:C2H2 zinc finger protein plays a key role in plant growth and development, abiotic stress response. Based on the previous work in laboratory, the subtractive hybridization library of root inhibition of Upland cotton under salt stress was analyzed, and a salt stress response gene GhC2H2. was screened out. The full length of the gene was cloned by RACE (Rapid amplification of cDNA ends) and RT-PCR (Reverse transcription-polymerase chain reaction), and its cellular localization was analyzed by transient expression analysis. The expression patterns of the gene in root, stem and leaf of upland cotton under salt stress were analyzed by Real time-PCR. The boll weight, lint percentage and fiber quality index of the transgenic lines in normal soil and saline soil were determined. The GenBank accession number of the gene was 819 bp, encoding 272 aa;. The encoded protein contained two C2H2 zinc-finger domains and was located in the nucleus. The expression of the gene was up-regulated in different tissues induced by salt stress, and the up-regulation was the highest in the early stage of stress induction. It was speculated that the gene was involved in the early salt stress response. The analysis of the characters of transgenic progenies showed that the overexpression of the gene could improve lint score and fiber quality of transgenic cotton. It was inferred that GhC2H2 was involved in both salt stress response and fiber development regulation.
【作者单位】： 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;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;湖北省农业科学院经济作物研究所;
【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)
【分类号】：S562
，
本文编号：2393206
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