棉花GhDMT3的功能验证及生物信息学分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0724

生物技术通报 ›› 2019, Vol. 35 ›› Issue (1): 11-16.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0724

棉花GhDMT3的功能验证及生物信息学分析

杨笑敏, 王俊娟, 王德龙, 陆许可, 陈修贵, 郭丽雪, 王帅, 陈超, 王晓歌, 韩明格, 叶武威

中国农业科学院棉花研究所棉花生物学国家重点实验室农业部棉花遗传改良重点开放实验室,安阳 455000

收稿日期:2018-08-13 出版日期:2019-01-26 发布日期:2019-01-23
作者简介:杨笑敏,女,硕士,研究方向：种质资源;E-mail：YXM18539738032@163.com
基金资助:
国家重点研发计划（2018YFD0100401）

Functional Verification and Bioinformatics Analysis of Cotton GhDMT3 Gene

YANG Xiao-min, WANG Jun-juan, WANG De-Long, LU Xu-ke, CHEN Xiu-gui, GUO Li-xue, WANG Shuai, CHEN Chao, WANG Xiao-ge, HAN Ming-ge, YE Wu-wei

Institute of Cotton Research,Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology/Key Laboratory for Cotton Genetic Improvement,Ministry of Agriculture,Anyang 455000

Received:2018-08-13 Published:2019-01-26 Online:2019-01-23

摘要/Abstract

摘要： 通过构建棉花GhDMT3的VIGS沉默载体并侵染棉花,探究该基因在棉花和高等植物中的功能。采用生物信息学方法对棉花DNA甲基转移酶基因GhDMT3（CotAD_46796）的DNA序列结构特性、该基因编码的蛋白质的理化性质、疏水性和蛋白二级结构等方面进行分析。克隆该基因,并构建pYL156：GhDMT3沉默载体。蛋白质相对分子质量为71 107.02 kD,等电点为4.85,不稳定系数为36.33。不同胁迫处理（冷、干旱）VIGS沉默GhDMT3的棉花幼苗表现出明显的表型差异。qRT-PCR的表达模式分析发现,不同胁迫下pYL156侵染的棉花中不同部位表达量没有变化,用pYL156：GhDMT3侵染过的棉花中GhDMT3转录水平与对照相比明显下降。与未沉默GhDMT3的棉花幼苗相比,沉默GhDMT3的棉花幼苗抗逆性显著增强。

关键词: 陆地棉, GhDMT3, 生物信息学分析, VIGS沉默

Abstract: The function of the gene in cotton and higher plants was explored by building VIGS silencing vector of GhDMT3 and infecting cotton. Bioinformatics was used to analyze the structure characteristics of DNA sequence of cotton DNA methyltransferase gene GhDMT3（CotAD_46796）as well as the physicochemical properties,hydrophobicity,and secondary structure of the protein encoded by this gene. The gene was cloned and the pYL156：GhDMT3 silencing vector was constructed. The relative molecular weight of the protein was 71 107.02 kD,the pI was 4.85,and the instability coefficient was 36.33. Cotton seedlings of VIGS silenced GhDMT3 treated with different stress（cold and dry）showed significant phenotypic differences. The expression pattern analysis of qRT-PCR showed that the expression levels in the different parts of pYL156-infected cotton did not vary under different stress,while the transcriptional Level of GhDMT3 in pYL156：GhDMT3-infected cotton decreased significantly compared with the control. Compared with cotton seedlings without GhDMT3 silenced,cotton seedlings with GhDMT3 silenced were significantly resistant to stress.

Key words: Gossypium hirsutum L., GhDMT3, bioinformatics analysis, VIGS silencing

杨笑敏, 王俊娟, 王德龙, 陆许可, 陈修贵, 郭丽雪, 王帅, 陈超, 王晓歌, 韩明格, 叶武威. 棉花GhDMT3的功能验证及生物信息学分析[J]. 生物技术通报, 2019, 35(1): 11-16.

YANG Xiao-min, WANG Jun-juan, WANG De-Long, LU Xu-ke, CHEN Xiu-gui, GUO Li-xue, WANG Shuai, CHEN Chao, WANG Xiao-ge, HAN Ming-ge, YE Wu-wei. Functional Verification and Bioinformatics Analysis of Cotton GhDMT3 Gene[J]. Biotechnology Bulletin, 2019, 35(1): 11-16.

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棉花GhDMT3的功能验证及生物信息学分析

Functional Verification and Bioinformatics Analysis of Cotton GhDMT3 Gene

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