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核盘菌自噬相关基因SsATG5SsATG8的功能分析
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引用本文:刘勇,张宇宇,付艳苹,谢甲涛,姜道宏,程家森.核盘菌自噬相关基因SsATG5SsATG8的功能分析.植物保护学报,2024,51(2):302-313
DOI:10.13802/j.cnki.zwbhxb.2024.2023049
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作者单位E-mail
刘勇 华中农业大学, 农业微生物学国家重点实验室, 武汉 430070
华中农业大学植物科学技术学院, 湖北省作物病害监测和安全控制重点实验室, 武汉 430070 
 
张宇宇 华中农业大学, 农业微生物学国家重点实验室, 武汉 430070
华中农业大学植物科学技术学院, 湖北省作物病害监测和安全控制重点实验室, 武汉 430070 
 
付艳苹 华中农业大学, 农业微生物学国家重点实验室, 武汉 430070
华中农业大学植物科学技术学院, 湖北省作物病害监测和安全控制重点实验室, 武汉 430070 
 
谢甲涛 华中农业大学, 农业微生物学国家重点实验室, 武汉 430070
华中农业大学植物科学技术学院, 湖北省作物病害监测和安全控制重点实验室, 武汉 430070 
 
姜道宏 华中农业大学, 农业微生物学国家重点实验室, 武汉 430070
华中农业大学植物科学技术学院, 湖北省作物病害监测和安全控制重点实验室, 武汉 430070 
 
程家森 华中农业大学, 农业微生物学国家重点实验室, 武汉 430070
华中农业大学植物科学技术学院, 湖北省作物病害监测和安全控制重点实验室, 武汉 430070 
jiasencheng@mail.hzau.edu.cn 
中文摘要:为探究自噬在核盘菌Sclerotinia sclerotiorum致病过程中的作用,利用酵母Saccharomyces自噬相关基因(autophagy-related gene,ATG)编码的蛋白序列比对核盘菌基因组,获得核盘菌假定ATG,并以核盘菌1980菌株为出发菌株,基于同源重组的原理对假定ATG进行敲除和回补,并测定不同突变体的生长表型和致病能力。结果表明,从核盘菌基因组中比对到2个ATG,分别命名为SsATG5SsATG8,两者在核盘菌致病过程中均上调表达。SsATG5SsATG8敲除突变体在菌丝生长、产草酸和侵染垫形成方面与野生型菌株无明显差异,但SsATG5敲除突变体在离体拟南芥Arabidopsis thaliana叶片上的致病力显著下降了约40%,在活体拟南芥植株上的致病力显著下降了约80%,同时SsATG5回补突变体恢复了正常的致病力。表明SsATG5参与了核盘菌的致病过程,证实自噬在核盘菌致病过程中发挥着重要作用。
中文关键词:核盘菌  SsATG5  SsATG8  自噬  致病力  侵染垫
 
Functional analysis of autophagy-related genes SsATG5 and SsATG8 in pathogenic fungus Sclerotinia sclerotiorum
Author NameAffiliationE-mail
Liu Yong State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China
Hubei Key Laboratory of Crop Disease Monitoring and Safety Control, School of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China 
 
Zhang Yuyu State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China
Hubei Key Laboratory of Crop Disease Monitoring and Safety Control, School of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China 
 
Fu Yanping State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China
Hubei Key Laboratory of Crop Disease Monitoring and Safety Control, School of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China 
 
Xie Jiatao State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China
Hubei Key Laboratory of Crop Disease Monitoring and Safety Control, School of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China 
 
Jiang Daohong State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China
Hubei Key Laboratory of Crop Disease Monitoring and Safety Control, School of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China 
 
Cheng Jiasen State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China
Hubei Key Laboratory of Crop Disease Monitoring and Safety Control, School of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China 
jiasencheng@mail.hzau.edu.cn 
Abstract:In order to explore the functions of autophagy in the pathogenesis of Sclerotinia sclerotiorum, the putative autophagy-related gene (ATG) was identified by homologous alignment of protein sequences of yeast autophagy-related genes. According to the principle of homologous recombination, the ATG was knocked out and then complemented in the S. sclerotiorum strain 1980, and then the phenotype of growth and virulence of different mutants were determined. The results showed that SsATG5 and SsATG8 were identified in S. sclerotiorum, and the expression of both of them were significantly upregulated during the infection of S. sclerotiorum. There was no significant difference between SsATG5 and SsATG8 knockout mutants and wild-type strain 1980 in mycelial growth, oxalic acid production and infection cushion formation. However, the virulence of SsATG5 knockout mutants significantly decreased by about 40% on the detached leaves of A. thaliana, while it significantly decreased by about 80% on A. thaliana plants. Importantly, the virulence of SsATG5 complementary strain was not different from that of the wild-type strain 1980. The results indicated that SsATG5 was involved in the pathogenesis of S. sclerotiorum, and suggested that the autophagy pathway played important roles in the pathogenesis of S. sclerotiorum.
keywords:Sclerotinia sclerotiorum  SsATG5  SsATG8  autophagy  virulence  infection cushion
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