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作物学报 ›› 2013, Vol. 39 ›› Issue (11): 1962-1969.doi: 10.3724/SP.J.1006.2013.01962

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

棉属野生种旱地棉苏氨酸醛缩酶基因GarTHA的克隆及功能验证

范昕琦,刘章伟,冯娟,徐鹏,张香桂,沈新莲*   

  1. 江苏省农业科学院经济作物研究所 / 农业部长江下游棉花与油菜重点实验室, 江苏南京210014
  • 收稿日期:2013-03-29 修回日期:2013-06-09 出版日期:2013-11-12 网络出版日期:2013-07-31
  • 通讯作者: 沈新莲, E-mail: xlshen68@126.com
  • 基金资助:

    本研究由国家转基因生物新品种培育重大专项(2011ZX08005-004-002)和江苏省农业科技自主创新基金项目(CX(11)1201)资助。

Cloning and Functional Analysis of GarTHA Gene from Gossypium aridum

FAN Xin-Qi,LIU Zhang-Wei,FENG Juan,XU Peng,ZHANG Xiang-Gui,SHEN Xin-Lian*   

  1. Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences / Key Laboratory of Cotton and Rapeseed (Nanjing), Ministry of Agriculture, Nanjing 210014, China
  • Received:2013-03-29 Revised:2013-06-09 Published:2013-11-12 Published online:2013-07-31
  • Contact: 沈新莲, E-mail: xlshen68@126.com

摘要:

盐胁迫是影响作物生长和发育的重要因素之一。一些棉属野生种具有较好的耐盐性, 是开展棉花耐盐性机制研究以及改良陆地棉耐盐性的重要资源。本研究基于cDNA-AFLP技术分离获得的旱地棉(Gossypium aridum)盐胁迫下差异表达片段序列信息, 经电子克隆技术和RT-PCR方法克隆了旱地棉苏氨酸醛缩酶基因cDNA全长, 命名为GarTHA (GenBank登录号为KC167360)。该cDNA全长1 018 bp, 包含一个822 bp的完整ORF, 编码273个氨基酸残基, 蛋白质分子量为82.57 kD, 等电点为4.89GarTHA基因与杨树PtTHA基因同源性最高, 84.6%为进一步验证其功能, 利用拟南芥逆境胁迫启动子rd29A构建植物表达载体, GarTHA基因的完整ORF转入拟南芥中, 获得转基因植株并进行了耐盐性鉴定。结果表明, 在盐胁迫下转基因拟南芥种子的发芽率明显高于野生型, 且转基因植株的根长显著高于野生型。说明GarTHA基因可能参与植物的盐胁迫反应, 从而提高植物抗逆性。

关键词: 旱地棉, 苏氨酸醛缩酶, 耐盐性, 结构特征, 功能分析

Abstract:

Salinity stress is one of the most important factors that impede the growth and development of various crops. Some Gossypium wild species with tolerance to high salinity are valuable germplasm resource for studying salt tolerance mechanism in Gossypium and improving salinity resistance in upland cotton. In the previous study, we obtained a differential expression fragment by comparing differential expression transcript under stress treatment in diploid G. aridum species using cDNA-AFLP technique. In this study, using the partial cDNA sequences as queries, the Gossypium EST database was screened and the corresponding cDNA sequence containing a complete ORF was assembled. As a result, a novel gene, encoding G. aridum threonine aldolase, was cloned. The gene was designated as GarTHA (G. aridum THA;GenBank accession number: KC167360). The ORF of GarTHA was 822 bp encoding 273 amino acid residues with a predicted molecular weight of 82.57 kD and a predicted isoelectric point of 4.89. The GarTHA showed the highest similarity of 84.6% with poplar PtTHA. To characterize its putative function, we transgenic the ORF of GarTHA gene driven by rd29A promoter into Arabidopsis. The growth of transgenic plants was observed under salinity stress. The seed germination rate and root length of transgenic Arabidopsis were significantly higher than those of the wild type plants under salt stress. The results showed that GarTHA gene could improve salt resistance in plants.

Key words: 旱地棉, 苏氨酸醛缩酶, 耐盐性, 结构特征, 功能分析

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