华北农学报 ›› 2018, Vol. 33 ›› Issue (6): 1-7. doi: 10.7668/hbnxb.2018.06.001

所属专题: 小麦

• 论文 • 上一篇    下一篇

普通小麦MYB转录因子Tamyb59的克隆及表达分析

张鹏钰1,2, 刘毓侠3, 曹丽茹1,2,3, 袁珍2, 王国瑞2, 王同朝1, 尹钧2,4, 卫丽2,4   

  1. 1. 河南农业大学 河南粮食作物协同创新中心, 河南 郑州 450002;
    2. 河南农业大学, 河南 郑州 450002;
    3. 河南省农业科学院, 河南 郑州 450002;
    4. 国家小麦工程技术研究中心, 河南 郑州 450002
  • 收稿日期:2018-08-20 出版日期:2018-12-28
  • 通讯作者: 卫丽(1966-),女,河南郑州人,研究员,博士,主要从事小麦发育研究;刘毓侠(1965-),女,河南商丘人,研究员,主要从事作物遗传育种及期刊编辑工作。
  • 作者简介:张鹏钰(1991-),女,河南郑州人,在读博士,主要从事小麦发育研究。
  • 基金资助:
    国家自然科学基金项目(31471452);河南省科学技术研究计划项目(182102110101);国家重点研究发展计划项目(2017YFD0301106)

Cloning and Expression Analysis of Transcription Factor Tamyb59 in Wheat

ZHANG Pengyu1,2, LIU Yuxia3, CAO Liru1,2,3, YUAN Zhen2, WANG Guorui2, WANG Tongchao1, YIN Jun2,4, WEI Li2,4   

  1. 1. Collaborative Innovation Center of Henan Food, Henan Agricultural University, Zhengzhou 450002, China;
    2. Henan Agricultural University, Zhengzhou 450002, China;
    3. Henan Academy of Agricultural Sciences, Zhengzhou 450002, China;
    4. The National Engineering Research Centre for Wheat, Zhengzhou 450002, China
  • Received:2018-08-20 Published:2018-12-28

摘要: 为进一步挖掘小麦逆境胁迫响应基因在植物非生物逆境胁迫应答中的作用,探究小麦逆境胁迫响应机制,从前期转录组结果中筛选出1个编码MYB蛋白的基因,暂命名为Tamyb59,通过PCR技术扩增Tamyb59的全长;利用NCBI和DNAMAN软件进行基因序列比对和保守结构域的分析;利用Expasy和TMHMM等在线软件进行氨基酸组成、亲水系数分析;采用MEGA 6.0软件构建系统进化树;构建pMDC83-GFP融合表达载体,进行基因表达的亚细胞定位分析。采用qRT-PCR分析了基因在不同非生物胁迫处理过程中不同组织的表达特性,并利用SAS数据处理软件进行差异显著性分析。结果表明:该基因含有典型的SANT保守结构域,CDS序列全长为522 bp,编码173个氨基酸,编码蛋白分子质量为19.7 ku,理论等电点为7.61。基因系统进化分析结果表明,Tamyb59与山羊草、粳稻、玉米等9种植物MYB转录因子有52.0%~85.6%的同源性,其中与谷子的MYB蛋白同源性最高。亚细胞定位结果显示,Tamyb59基因编码蛋白定位在细胞核。利用qRT-PCR分析了基因在不同非生物胁迫处理过程中不同组织的表达特性,组织表达特异性分析显示,Tamyb59基因在小麦根中的表达量较高,茎、叶和幼穗中表达量较低;PEG和盐胁迫处理过程中,Tamyb59基因的表达均呈现先上升后下降的趋势,说明Tamyb59基因对不同非生物胁迫有不同的响应。

关键词: 小麦, Tamyb59基因, 基因克隆, 非生物胁迫, 表达分析

Abstract: In order to further excavation of the role of wheat stress response genes in plant abiotic stress response,explored the regulation mechanism of abiotic stress response, a MYB gene was screened from a transcriptome result and named Tamyb59, temporarily.The full length sequence of Tamyb59 was cloned by PCR. Sequence comparison and conserved domain were analyzed by NCBI and DNAMAN. Amino acid composition and hydrophilic coefficient were analyzed by online software Expasy and TMHMM. Phylogenic tree was constructed by MEGA 6.0 according to the NJ method. And a fusion expression vector pMDC83-GFP was constructed to identify the subcellular localization. The Real-time PCR (qRT-PCR) was used to analyze the expression characteristics of different tissues in different abiotic stress treatments. The significance was analyzed by SAS. The results showed that:Tamyb59 gene contained a typical conserved SANT domain. The full-length CDS of Tamyb59 was 522 bp,which encoded 173 amino acids with a molecular weight of 19.7 ku and the theoretical isoelectric point was 7.61;Phylogenetic analysis showed that Tamyb59 had 52.0%-85.6% homology with MYB transcription factors of 9 other plants (Aegilops tauschii, Oryza sativa Japonica and Zea may, etc.). Tamyb59 had the highest homology with the MYB sequence of Setaria italica. The protein of Tamyb59 encoded specifically located in the nucleus.qRT-PCR was used to analyze the expression characteristics of different tissues in different abiotic stress treatments. The results revealed that the expression level of Tamyb59 was the highest in the root, and lower in stem, leaf and young spike. Under the PEG and NaCl treatment stress, the expression of Tamyb59 showed a trend of rising first and then decreasing, indicating that Tamyb59 had different responses under different stresses.

Key words: Wheat, Tamyb59 gene, Gene cloning, Abiotic stress, Expression analysis

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引用本文

张鹏钰, 刘毓侠, 曹丽茹, 袁珍, 王国瑞, 王同朝, 尹钧, 卫丽. 普通小麦MYB转录因子Tamyb59的克隆及表达分析[J]. 华北农学报, 2018, 33(6): 1-7. doi: 10.7668/hbnxb.2018.06.001.

ZHANG Pengyu, LIU Yuxia, CAO Liru, YUAN Zhen, WANG Guorui, WANG Tongchao, YIN Jun, WEI Li. Cloning and Expression Analysis of Transcription Factor Tamyb59 in Wheat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(6): 1-7. doi: 10.7668/hbnxb.2018.06.001.

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