玉米大斑病菌转录因子StbHLH10基因的克隆及其功能分析  

Cloning and Functional Analysis of Transcription Factor Gene StbHLH10 in Setosphaeria turcica

作　　者：韩丹 张心雨 王小敏 肖怡铭 裴智勇[3] 谷守芹[1] 刘玉卫 巩校东[1] HAN Dan;ZHANG Xin-Yu;WANG Xiao-Min;XIAO Yi-Ming;PEI Zhi-Yong;Gu Shou-Qin;LIU Yu-Wei;GONG Xiao-Dong(College of Life Sciences//Hebei Agricultural Microbial Bioinformation Utilization Technology Innovation Center/Hebei Key Laboratory of Plant Physiology and Molecular Pathology,Hebei Agricultural University,Baoding 071001,China;College of Modern Science and Technology,Hebei Agricultural University,Baoding 071001,China;Biocomputing Department of Beijing Computing Centre,Beijing 100094,China)

机构地区：[1]河北农业大学生命科学学院/河北省农业微生物生物信息利用技术创新中心/河北省植物生理与分子病理学重点实验室,保定071001 [2]河北农业大学现代科技学院,保定071001 [3]北京市计算中心生物计算事业部,北京100094

出　　处：《农业生物技术学报》2024年第5期1132-1140,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31671983);中央引导地方科技发展资金(216Z2902G);现代农业产业技术体系专项(CARS-02)。

摘　　要：碱性螺旋-环-螺旋(basic helix-loop-helix,bHLH)转录因子家族广泛参与植物病原真菌的生长发育和侵染致病过程。StbHLH10是玉米大斑病菌(Setosphaeria turcica)中编码bHLH类转录因子的基因,为了解该基因的功能,本研究以玉米大斑病菌为材料克隆了StbHLH10(GenBank No.XP_0080332037.1)CDS全长,并对其序列和结构特征进行了分析。结果显示,该基因CDS全长1659 bp,编码552个氨基酸,其编码的蛋白质序列在第335~441位氨基酸残基处含有1个HLH保守结构域,由2个α-螺旋和1个Loop环构成特定空间结构;表达模式分析显示,StbHLH10基因的表达量随着病菌侵染玉米叶片的时间延长而增加;通过构建其调控网络发现,StbHLH10可能通过调控33个下游靶基因的表达参与病菌的生长发育和侵染过程;原核诱导表达分析发现,在28℃诱导6 h条件下StbHLH10-His融合蛋白在大肠杆菌(Escherichia coli)中高效表达,分子量为78.5 kD,与理论值一致,并通过镍柱亲和层析获得该转录因子可溶性的纯化蛋白。本研究初步探究了StbHLH10转录因子的功能,为进一步研究该基因的作用机制提供数据支持。As a conserved transcription factor family,basic Helix-Loop-Helix(bHLH)is widely involved in the development and infection of plant pathogenic fungi.StbHLH10 is a member of bHLH transcription factor family in Setosphaeria turcica.In order to understand the function of this gene,the full-length CDS of StbHLH10(GenBank No.XP_008030228.1)was cloned from S.turcica,and the structural characteristics of its protein sequence were analyzed.The results showed that the CDS of the StbHLH10 was 1659 bp in length,encoding 552 amino acids.The protein sequence of StbHLH10 contained a HLH conserved domain at residues 335~441 aa,which formed a spatial structure of protein by 2α-helices and a loop ring.The expression pattern analysis showed that the expression level of StbHLH10 gene increased with the prolongation of the time that the pathogen infected maize leaves.By constructing its regulatory network,it was found that the StHLH10 may be involved in the development and infection of S.turcica by regulating the expression of 36 downstream target genes.Prokaryotic expression analysis showed that the StbHLH10-His fusion protein was highly expressed in Escherichia coli at 28℃for 6 h,and the molecular weight was 78.5 kD,which was consistent with the theoretical value.The soluble purified protein of StbHLH10 was obtained by nickel column affinity chromatography.This study preliminarily explores the function of StbHLH10 transcription factor,and provides reference for further research on the mechanism of StbHLH10.

关 键 词：玉米大斑病菌 转录因子bHLH StbHLH10基因 侵染表达 原核表达 

分 类 号：S435.131[农业科学—农业昆虫与害虫防治]