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作物学报 ›› 2017, Vol. 43 ›› Issue (08): 1181-1189.doi: 10.3724/SP.J.1006.2017.01181

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

木薯转录因子基因MeHDZ14的克隆与分析

于晓玲,阮孟斌,王斌,杨义伶,王树昌*,彭明*   

  1. 中国热带农业科学院热带生物技术研究所 / 农业部热带作物生物学与遗传资源利用重点实验室, 海南海口 571101
  • 收稿日期:2016-12-09 修回日期:2017-04-20 出版日期:2017-08-12 发布日期:2017-05-11
  • 通讯作者: 王树昌, E-mail: wangshuchang2001@163.com; 彭明, E-mail: pengming@itbb.org.cn E-mail:yuxiaoling@itbb.org.cn
  • 基金资助:

    本研究由国际科技合作与交流项目(31561143012, 2013DFA32020)和国家青年科学基金项目(31501378)资助。

Cloning and Analysis of Structure and Expression of MeHDZ14 Gene in Cassava

YU Xiao-Ling,RUAN Meng-Bin,WANG Bin,YANG Yi-Ling,WANG Shu-Chang*,PENG Ming*   

  1. Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Science / Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Haikou 571101, China
  • Received:2016-12-09 Revised:2017-04-20 Online:2017-08-12 Published:2017-05-11
  • Contact: 王树昌, E-mail: wangshuchang2001@163.com; 彭明, E-mail: pengming@itbb.org.cn E-mail:yuxiaoling@itbb.org.cn
  • Supported by:

    This study was supported by the International Science & Technology Cooperation Program of China (31561143012,2013DFA32020) and the National Natural Science Foundation of China (31501378).

摘要:

HD-Zip家族基因在植物生长发育和逆境胁迫中起重要作用。为了研究MeHDZ14基因在非生物胁迫(尤其是干旱)应答中的作用,选用对干旱信号反应灵敏、相对耐旱的木薯品种“SC124”做为实验材料,利用RT-PCR克隆了MeHDZ14基因。生物信息学分析发现,MeHDZ14基因编码的蛋白具有典型的HD-Zip保守结构域。将该基因编码的蛋白与GFP融合,亚细胞MeHDZ14:GFP重组蛋白定位于细胞核。同时,酵母Y187中的转录自激活试验结果也表明,MeHDZ14蛋白具有明显转录自激活功能。推断MeHDZ14是一个典型的HD-Zip I转录因子。MeHDZ14启动子区具有多个ABA响应元件ABRE (ABA response element);基因差异表达分析结果表明,MeHDZ14基因在叶片和根中的表达受干旱胁迫的诱导,并对外源ABA具有明显的响应。因此,认为MeHDZ14基因通过ABA依赖信号传导途径参与调控木薯干旱响应。此外,还发现MeHDZ14基因的编码区虽然存在数个SNP,但表现出高度保守性;且在不同木薯品种中的表达对干旱胁迫均有明显的响应,为进一步研究该基因的功能奠定了基础。

关键词: 木薯, MeHDZ14, 结构特性, 表达分析

Abstract:

HD-Zip family genes play an important role in plant growth and stress response. To reveal the role of MeHDZ14 gene in abiotic stresses (e.g. drought) in cassava, we cloned MeHDZ14 gene by using RT-PCR from cassava cultivar SC124, which was relatively more resistant to drought stress. Bioinformatics methods were used to analyze its structural characteristics, and semi-RT-PCR/qRT-PCR was used to explore its expression patterns in response to abiotic stresses in different plant tissues and varieties. MeHDZ14 has a 726 bp open reading frame, encoding 241 amino acids, and contains the typical HD and ZIP domain. Blastp analysis showed that MeHDZ14 has close genetic relationship with ATHB-7, which is a member of the family I HD-Zip gene. Yeast and subcellular localization test showed that the MeHDZ14 gene is a transcription factor and specifically expresses in the nucleus. Genetic structural variation analysis revealed a total of four mis-sense mutations in eight tested varieties. However, amino acid mutations were not found between wild and cultivated cassavas. This indicates the MeHDZ14 proteins are highly conserved. Semi-RT PCR analysis revealed that MeHDZ14 was specifically expressed in petioles, and induced by drought stress in root and leaf, suggesting that MeHDZ14 plays an important role in the early drought stage. Analysis by qRT-PCR showed that MeHDZ14 gene had different expression levels in different cassava varieties, but the same mode under drought stress and ABA treatment. These data indicate that MeHDZ14 is a member of the ABA pathway responding to drought. Our results showed that MeHDZ14 plays an important role in the molecular pathways of cassava drought resistance, underlining its potential in genetic improvement of cassava drought tolerance.

Key words: Cassava, MeHDZ14, Structure characteristics, Expression analysis

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