苹果树腐烂病菌异柠檬酸裂解酶基因VmICL的功能分析  

Functional study of isocitrate lyase gene VmICL in Valsa mali

作　　者：刁雨菲 于成明[1] 熊雄 郑金柱 靳纪洋 刘会香[1] DIAO Yufei;YU Chengming;XIONG Xiong;ZHENG Jinzhu;JIN Jiyang;LIU Huixiang(Shandong Research Center for Forestry Harmful Biological Control Engineering and Technology,College of Plant Protection/Shandong Agricultural University,Tai’an 271018,China;College of forestry/Shandong Agricultural University,Tai’an 271018,China;Culai Mountain Forest Farm,Tai’an 271027,China)

机构地区：[1]山东农业大学植物保护学院/山东省林业有害生物防控工程技术研究中心,泰安271018 [2]山东农业大学林学院,泰安271018 [3]徂徕山林场,泰安271027

出　　处：《植物病理学报》2023年第6期1120-1130,共11页Acta Phytopathologica Sinica

基　　金：国家自然基金项目(31770684);山东省自然科学基金项目(ZR2017MC042)。

摘　　要：由黑腐皮壳菌(Valsa mali Miyabe et Yamada)引起的苹果树腐烂病严重影响了苹果树的健康生长。异柠檬酸裂解酶是真菌乙醛酸循环中的关键酶,异柠檬酸裂解酶基因(ICL)在真菌生长发育及致病过程中发挥重要作用。本研究在苹果树腐烂病菌基因VmICL生物信息学分析的基础上,应用PEG介导原生质体转化法获得了基因VmICL的敲除突变体及回补菌株,解析了VmICL对病菌生长发育、细胞壁完整性、渗透胁迫、碳氮源利用及致病力的影响。研究表明,基因VmICL位于9号染色体,VmICL蛋白在第23-545位氨基酸区域具TIM super family结构域,系统发育分析显示与水稻稻瘟病菌(Magnaporthe grisea)亲缘关系最近。基因VmICL缺失突变体生长速率减慢,分生孢子器产生数量减少,分生孢子萌发延迟;基因VmICL参与维持细胞壁的完整性,但不参与渗透胁迫;基因VmICL正调控碳源吸收,负调控氮源吸收,正调控致病力。总之,基因VmICL参与调控苹果树腐烂病菌的生长发育、细胞壁完整性的维持、碳氮源利用及致病力的发挥。Valsa mali is caused by Valsa mali Miyabe et Yamada has seriously endangers the healthy growth of apple tree.Iocitrate lyase is a key enzyme in the fungal glyoxylic acid cycle,and the isocitrate lyase gene(ICL)plays an important role in the fungal growth and development and pathogenic processes.In this study,based on the bioinformatics analysis of the gene VmICL.VmICL deletion mutants and complement strains were obtained by PEG-mediated protoplast transformation.The effects of VmICL on pathogen growth and development,cell wall integrity,osmotic stress,carbon and nitrogen source utilization,and pathogenicity were analyzed.The results show that the gene VmICL is located on chromosome 9,and the VmICL protein has a TIM super family domain at amino acids positions 23-545.Phylogenetic analysis shows a recent relationship to Magnaporthe grisea.The VmICL deletion mutant had a slower growth rate,a lower number of pycnidia,and adelayed conidial germination.Gene VmICL is involved in maintaining the integrity of the cell wall,but was not involved in osmotic stress.Gene VmICL positive regulation for carbon source utilization,negative regulation for nitrogen source utilization,and positively regulating the pathogenicity.In conclusion,the VmICL is involved in the regulation of mycelial growth,pycnidia production and conidial germination,cell wall integrity maintenance,carbon and nitrogen source utilization,and pathogenicity of Valsa mali.

关 键 词：苹果树腐烂病菌 异柠檬酸裂解酶 VmICL 生物学功能 

分 类 号：S432.1[农业科学—植物病理学]