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棉花学报 ›› 2017, Vol. 29 ›› Issue (2): 138-146.doi: 10.11963/issn.1002-7807.201702003

• 研究与进展 • 上一篇    下一篇

陆地棉NAC转录因子基因GhNAC6的克隆、表达和耐盐性分析

杨勇(),田新权(),刘会利,宋洁,张慧,龙璐,高巍()   

  1. 河南大学生命科学学院/棉花生物学国家重点实验室/河南省植物逆境生物学重点实验室,河南 开封 475004
  • 收稿日期:2016-04-20 出版日期:2017-03-15 发布日期:2017-03-15
  • 通讯作者: 高巍 E-mail:283437814@qq.com;821022205@qq.com;gaowei021@163.com
  • 作者简介:杨勇(1985-),男,硕士,283437814@qq.com;田新权(1990-),男,硕士,821022205@qq.com;#同等贡献
  • 基金资助:
    棉花生物学国家重点实验室开放课题(CB2015A31,CB2016A23);河南省教育厅项目(15A180028, 15A180029);河南大学科学研究基金(2015YBZR030,2015YBZR031)

Cloning, Expression and Salt-Tolerance Analysis of the NAC Transcription Factor Gene GhNAC6 in Upland Cotton

Yong Yang(),Xinquan Tian(),Huili Liu,Jie Song,Hui Zhang,Lu Long,Wei Gao()   

  1. School of Life Science, Henan University / State Key Laboratory of Cotton Biology/Henan Key Laboratory of Plant Stress Biology, Kaifeng, Henan 475004, China
  • Received:2016-04-20 Online:2017-03-15 Published:2017-03-15
  • Contact: Wei Gao E-mail:283437814@qq.com;821022205@qq.com;gaowei021@163.com

摘要: 【目的】NAC(NAM-ATAF-CUC)家族是植物细胞内特有的1类转录因子家族,参与植物的抗逆过程。为了研究棉花抗逆的分子机制,本研究从陆地棉TM-1中克隆到1个NAC转录因子基因并研究了其功能。【方法】根据其序列同源性和进化分析结果,将该基因命名为GhNAC6,本文对GhNAC6进行了生物信息学和表达分析,并利用病毒诱导基因沉默技术在棉花中沉默GhNAC6的表达。【结果】GhNAC6全长为1629 bp,包含2个内含子,其中编码区为900 bp,编码1个相对分子质量为33.9×103、等电点为6.18的蛋白。GhNAC6启动子区段包含多个与诱导表达和组织特异表达相关的顺式作用元件。转录组数据分析结果表明,GhNAC6在棉花发育的不同时期具有时空表达差异性;实时荧光定量核酸扩增检测结果显示GhNAC6还受到植物激素水杨酸、乙烯利、茉莉酸甲酯和逆境胁迫低温、高温、高盐、伤口的诱导。干涉GhNAC6基因发现,降低GhNAC6基因的表达能增强棉花对盐胁迫的耐受性。【结论】研究结果表明GhNAC6可能参与了棉花发育、逆境响应和激素信号传导的过程。

关键词: 棉花; NAC转录因子; 表达模式; 激素; 盐胁迫

Abstract:

[Objective] NAC (NAM-ATAF-CUC) family members are plant-specific transcription factors involved in plant resistance to stress. To understand the molecular mechanism of cotton resistance to stress, we cloned and functional analyzed a NAC transcription factor gene from upland cotton cultivar TM-1. [Method] This gene was named GhNAC6 in accord with the results of a sequence similarity search and phylogenetic analysis. GhNAC6 was go through the bioinformatics and expression analysis, and further silenced in cotton using the virus-induced gene silencing system in this study. [Result] The full-length sequence of GhNAC6 was found to comprise 1629 bp, including two introns, and its 900-bp coding sequence was predicted to encode a protein with a relative molecular mass of 33.9 kDa and an isoelectric point of 6.18. The GhNAC6 promoter region was cloned and analyzed, which revealed the existence of several potential cis-elements involved in induced and tissue-specific expression. Transcriptome and qPCR analyses further revealed that GhNAC6 was differentially expressed in different tissues and developmental stages of cotton and was induced by various phytohormones (salicylic acid, ethephon and methyl jasmonate) and stresses (cold, heat, salinity and wounding). The GhNAC6-silenced cotton seedlings showed more tolerance to salt stress. [Conclusion] These data indicate that GhNAC6 may participate in cotton development, stress response and hormone signal transduction.

Key words: cotton; NAC transcription factor; expression profile; hormone; salt stress