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作物学报 ›› 2011, Vol. 37 ›› Issue (08): 1389-1397.doi: 10.3724/SP.J.1006.2011.01389

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦TaCRF2基因的克隆及其在烟草中的初步功能验证

郭丽香1,2,高世庆2,**,唐益苗2,王永波2,刘美英3,张朝3,徐蓓1,2,连微微1,2,赵昌平2,*   

  1. 1 首都师范大学生命科学学院, 北京100048; 2 北京市农林科学院杂交小麦工程技术研究中心, 北京100097; 3 中国科学技术大学研究生院化学系, 安徽合肥 230026
  • 收稿日期:2010-12-11 修回日期:2011-03-06 出版日期:2011-08-12 网络出版日期:2011-05-11
  • 通讯作者: 赵昌平, E-mail: zhaochangping401@vip.sohu.com, Tel: 13601096398
  • 基金资助:

    本研究由国家转基因生物新品种培育重大专项(2008ZX08002-002)，北京市自然科学基金项目(5102016)，北京市科技新星计划项目(2007B056，2008B035)和北京农林科学院院青年基金项目资助。

TaCRF2 Gene Isolation from Triticum aestivum and Primary Function Validation in Tobacco (Nicotiana tabacum L.)

GUO Li-Xiang1,2,GAO Shi-Qing2,**,TANG Yi-Miao2,WANG Yong-Bo2,LIU Mei-Ying3,ZHANG Zhao3,XU Bei1,2,LIAN Wei-Wei1,2,ZHAO Chang-Ping2,*   

  1. 1 College of Life Sciences, Capital Normal University, Beijing 100048, China; 2 Beijing Hybrid Wheat Engineering and Technology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China; 3 Department of Chemistry, Graduate School of University of Science and Technology of China, Hefei 230026, China
  • Received:2010-12-11 Revised:2011-03-06 Published:2011-08-12 Published online:2011-05-11
  • Contact: 赵昌平, E-mail: zhaochangping401@vip.sohu.com, Tel: 13601096398

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被引次数

6

摘要： 通过RT-PCR从小麦cDNA中扩增获得一个锌指蛋白基因TaCRF2, 该基因的cDNA长度为847 bp, 序列分析表明它编码一个含有280个氨基酸的蛋白质。在线软件预测该蛋白质的相对分子质量为30.97 kD, 等电点为7.03, 且在C-末端含有一个典型的RING-H2型锌指蛋白结构域, 在N-末端含有两个跨膜结构域。氨基酸序列比对发现, TaCRF2与水稻中的一个RING型锌指蛋白(ABF95226)的相似度为82%。亚细胞定位分析显示, 该蛋白分布在细胞核和细胞膜上。Real-time PCR表达特性分析显示, TaCRF2基因的表达受干旱、盐和ABA的强烈诱导, 低温对该基因的表达量影响不明显。初步功能验证发现过表达TaCRF2基因增强了转基因烟草对干旱和盐胁迫的耐性。

关键词: 烟草, 小麦, RING-H2型锌指蛋白, TaCRF2基因, 干旱胁迫, 盐胁迫

Abstract: Wheat, like other crops, suffers from drought, salt, low-temperature and other abiotic stresses, seriously resulting in yield decline. It is an effective way for crop breeding to improve resistance via molecular biology techniques. Zinc finger protein, an important transcription factor commonly found in plant, can regulate the expression of multiple stress-inducible genes and enhance comprehensive resistances effectively. In this study, a RING-H2 zinc ?nger protein gene, designated as TaCRF2, was isolated from Triticum aestivum by RT-PCR. Its cDNA was 847 bp and encoded a putative protein of 280 amino acids with a predicted molecular mass of 30.97 kD and an isoelectric point (PI) of 7.03. A typical RING-H2 finger domain was found at the C-terminal region of TaCRF2 protein, and two transmembrane domains were found at the N-terminal region. Alignment of amino acid sequence showed that TaCRF2 was 82% identical to Oryza sativa putative RING zinc finger protein ABF95226. Subcellular localization analysis showed that the TaCRF2 was expressed in both nuclear and cytoplasm membrane. Real-time PCR showed that the transcript of TaCRF2 was strongly induced by drought, salinity and cold to some extent. The TaCRF2 gene was transformed into tobacco cultivar Wisconsin 38 (W38) by Agrobacterium mediated under the control of the CaMV 35S promoter. Under drought, salt and low-temperature stresses, transgenic tobacco lines carrying TaCRF2 gene developed the strong primary root, more lateral root and smaller yellow leaves, and performed higher tolerance to these stresses than the wide-type tobaccos, The primary function verification showed that overexpression of TaCRF2 gene in tobaccos enhanced their tolerance to drought and salinity stresses. This study will help further research of wheat RING-H2 type zinc finger protein reactions in the role of stress resistance, and bring wheat resistance breeding excellent candidate genes.

Key words: Tobacco, Triticum aestivum, RING-H2 zinc-finger protein, TaCRF2 gene, Drought stress, Salinity stress

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