棉花抗枯萎病相关基因GhWRKY48的克隆及表达分析  

Cloning and Expression Analysis of Cotton Blight Resistance-Related Gene GhWRKY48

作　　者：明聪 赵龙飞 刘戈辉 张薇[1] Ming Cong;Zhao Longfei;Liu Gehui;Zhang Wei(College of Agriculture,Shihezi University,Shihezi,832003)

机构地区：[1]石河子大学农学院,石河子832003

出　　处：《分子植物育种》2021年第11期3523-3530,共8页Molecular Plant Breeding

基　　金：国家自然科学基金(31260358)资助。

摘　　要：棉花枯萎病是影响棉花产量和品质的最重要病害,WRKY转录因子在植物抗病中扮演重要角色。本实验以高抗枯萎病的陆地棉‘中棉所12’为实验材料,以枯萎病菌诱导棉花根部基因表达谱中筛选得到WRKY基因片段为探针,从陆地棉‘中棉所12’根部c DNA中克隆WRKY转录因子基因。通过多序列比对分析发现该基因属于第Ⅱ类WRKY转录因子家族,将该基因命名为GhWRKY48。GhWRKY48基因的序列分析发现,cDNA全长为1074个核苷酸,编码了357个氨基酸,蛋白分子量为3.94 kD,脂肪指数为56.86,等电点6.20,含有1个WRKY结构域及1个C2H2型锌指属于疏水蛋白。基因的蛋白二级结构包括延伸链、α螺旋和无规则卷曲。通过RT-qPCR的结果发现,枯萎病菌处理后,GhWRKY48的基因表达量显著低于对照组,预测该基因可能参与棉花抗枯萎病反应体系。本研究结果为棉花抗枯萎病提供一定的研究思路与途径。Cotton Fusarium wilt is the most important disease affecting cotton yield and quality.The WRKY transcription factor also plays an important role in plant resistance.In this experiment,the upland cotton’Zhongmiansuo 12’,which is highly resistant to wilt,was used as the experimental material.The WRKY gene fragment was screened from the gene expression profile of the cotton root induced by Fusarium oxysporum.Cloned WRKY transcription factor gene.Through multiple sequence alignment analysis,it was found that the gene belongs to the WRKY transcription factor family of typeⅡ,and the gene was named GhWRKY48.Sequence analysis of the GhWRKY48 gene found that the full length of the cDNA was 1074 nucleotides,encoding 357 amino acids,the protein molecular weight is 3.94 kD,fat index is 56.86,isoelectric point is 6.20,contained a WRKY domain and a C2 H2 type zinc fingers are hydrophobins.The protein secondary structure of the gene consists of extended chains,alpha helices,and random coils.The results of RT-qPCR showed that the gene expression of GhWRKY48 was significantly lower than that of the control group after Fusarium oxysporum treatment.It is predicted that this gene may be involved in the cotton blight resistance system.The results of this study provide certain research ideas and ways for cotton resistance to Fusarium wilt.

关 键 词：生物学分析 基因克隆 枯萎病 WRKY转录因子 实时荧光定量PCR 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]