核农学报2025，Vol.39Issue(11)：2342-2350,9.DOI:10.11869/j.issn.1000-8551.2025.11.2342
玉米大斑病菌纤维素酶基因StCEL1克隆表达及其编码蛋白致病性分析
Cloning,Expression and Pathogenicity of StCEL1 Cellulase Gene in Setosphaeria turcica
摘要
Abstract
To investigate the function of cellulase and its key encoding genes in the pathogenic process of Setosphaeria turcica,the cellulase gene StCEL1 was cloned from the isolate YD001 and subjected to bioinformatic analysis.The recombinant plasmid pPIC9K/StCEL1 was constructed and successfully expressed in Pichia pastoris.Subsequently,enzymatic properties and pathogenicity of the recombinant protein were analyzed.The results showed that the StCEL1 gene was 1 308 bp in length,contained one intron,and encoded a 410-amino-acid protein with a molecular weight of 44.07 kDa and a theoretical isoelectric point of 8.19.The protein was hydrophilic,had a signal peptide,and lacked transmembrane structures.Domain and secondary structure analysis revealed that the StCEL1 protein contained a distinct conserved domain,belonged to the glycoside hydrolase 7-cellobiohydrolase-endoglucosidase superfamily,and exhibited the highest content of random coils in its secondary structure.The expression of StCEL1 in Pichia pastoris produced a recombinant protein with a molecular weight of approximately 66 kDa.Optimal expression conditions involved induction with 1.0%methanol for 96 h.The enzyme exhibited maximal activity at 55℃,pH 4.0,and a reaction duration of 30 min.Maize seedlings inoculated with this protein developed distinct disease lesions,suggesting that StCEL1 gene might contribute to the pathogenicity of S.turcica.These results provide a theoretical basis for further exploring the pathogenic mechanisms of StCEL1 protein.关键词
玉米大斑病菌/纤维素酶/基因克隆/毕赤酵母/致病性Key words
Setosphaeria turcica/cellulase/gene cloning/Pichia pastoris/pathogenicity引用本文复制引用
马周杰,张楠,高增贵,温胜慧,杨俊伟,王建军,赵变平..玉米大斑病菌纤维素酶基因StCEL1克隆表达及其编码蛋白致病性分析[J].核农学报,2025,39(11):2342-2350,9.基金项目
山西省基础研究计划项目(202303021222047),山西省科技重大专项计划揭榜挂帅项目子课题(202201140601025-1-02),忻州市基础研究计划项目(20220506),山西农业大学生物育种工程项目(YZGC064) （202303021222047）
