玉米大斑病菌转录因子StMR1调控的候选靶基因筛选及功能预测  

Screening and Functional Prediction of Candidate Target Genes Regulated by Transcription Factor StMR1 of Setosphaeria turcica

作　　者：尹玉娟 李海笑 王秋月 贾慧 刘宁[1,2] 董金皋 曹志艳[1,2] YIN Yu-Juan;LI Hai-Xiao;WANG Qiu-Yue;JIA Hui;LIU Ning;DONG Jin-Gao;CAO Zhi-Yan(College of Plant Protection,Hebei Agricultural University,Baoding 071000,China;State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology,Baoding 071000,China)

机构地区：[1]河北农业大学植物保护学院,保定071000 [2]华北作物改良与调控国家重点实验室/河北省植物生理与分子病理学重点实验室,保定071000

出　　处：《农业生物技术学报》2022年第12期2396-2406,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32072370);河北省自然科学基金(C2021204136,C2020204183);现代农业产业技术体系专项资金(CARS-02);大学生创新创业训练计划项目(2020316)。

摘　　要：玉米大斑病菌(Setosphaeria turcica)黑色素调控转录因子1(S.turcica melanin regulation factor 1,StMR1)具有双锌指结构域,通过调控基因表达影响玉米大斑病菌的生长、代谢和致病性,因此,阐明该转录因子的调控模式对解析玉米大斑病菌的致病机理至关重要。本课题组前期研究发现,StMR1基因被敲除后突变体黑色素合成受阻,菌落由黑色变为灰白色,致病力会显著降低。本研究利用玉米大斑病菌野生型菌株和StMR1基因缺失突变体的转录组测序(RNA sequence,RNA-seq)数据筛选差异表达基因,并对其进行基因本体论(Gene Ontology,GO)和KEGG功能注释分析,共筛选获得了1383个上调表达基因和1710个下调表达基因,下调表达基因与分子功能、细胞成分和生物学过程相关,涉及6类25条代谢通路。利用qRT-PCR对筛选获得的6个显著下调表达的基因SETTUDRAFT_172122、SETTUDRAFT_168776、SETTUDRAFT_166944、SETTUDRAFT_162601、SETTUDRAFT_131269和SETTUDRAFT_163249进行了表达量分析,结果发现,6个基因在突变体中的表达水平均呈下调趋势,说明转录因子StMR1对其可能具有正调控作用。生物信息学分析发现,SETTUDRAFT_162601基因具有典型的Catalase结构域,并通过qRTPCR检测到该基因在H_(2)O_(2)胁迫下表达量显著升高(P<0.05),推测该基因与抗氧化胁迫有关。上述研究结果为深入分析转录因子StMR1调控玉米大斑病菌致病性的机制提供了借鉴,也为解析锌指类转录因子功能提供参考。Setosphaeria turcica melanin regulation factor 1(StMR1)is a melanin transcriptional regulatory factor with a double zinc finger domain in Setosphaeria turcica,which affects the growth,metabolism and pathogenicity by regulating gene expression.Therefore,it is very important to elucidate the regulation mode of this transcription factor for analyzing the pathogenic mechanism of S.turcica.The previous study of our group found that after the StMR1 gene was knocked out,the synthesis of melanin in the mutant was blocked,the colony changed from black to gray-white,and the pathogenicity was significantly reduced.In this study,the RNA sequence(RNA-seq)data of wild-type strains of S.turcica and StMR1 gene deletion mutants were used to screen differentially expressed genes(GEGs).Gene Ontology(GO)and KEGG function annotation analysis were conducted for DEGs.The screening obtained 1383 up-regulated genes and 1710 downregulated genes.The down-regulated genes were related to molecular functions,cellular components and biological processes,involving 25 metabolic pathways in 6 categories.Six down-regulated genes,SETTUDRAFT_172122,SETTUDRAFT_168776,SETTUDRAFT_166944,SETTUDRAFT_162601,SETTUDRAFT_131269 and SETTUDRAFT_163249,were analyzed of the expression level by qRT-PCR.The qRT-PCR results showed that the expression levels of the 6 genes were down-regulated in the mutant,suggested that StMR1 might have a positive regulation effect on the genes.Bioinformatics analysis showed that gene SETTUDRAFT_162601 had a typical Catalase domain,and the expression level of SETTUDRAFT_162601 was significantly increased under H_(2)O_(2) by qRT-PCR(P<0.05),suggested that this gene was related to antioxidant stress.These results provide reference for further study on the mechanism of transcription factor StMR1 regulating pathogenicity,and also provide a reference for analyzing the function of zinc finger transcription factors.

关 键 词：玉米大斑病菌 黑色素 转录因子 黑色素调控转录因子1(MR1) 

分 类 号：S513[农业科学—作物学]