核盘菌自噬相关基因Ss4TG5和SsATG8的功能分析
Functional analysis of autophagy-related genes SsATG5 and SsATG8 in pathogenic fungus Sclerotinia sclerotiorum
刘勇 1张宇宇 1付艳苹 1谢甲涛 1姜道宏 1程家森1
作者信息
- 1. 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
- 折叠
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SsATG5和SsATG8,两者在核盘菌致病过程中均上调表达.SsATG5和SsATG8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.
Abstract
In order to explore the functions of autophagy in the pathogenesis of Sclerotinia sclerotio-rum,the putative autophagy-related gene(ATG)was identified by homologous alignment of protein se-quences 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 pheno-type 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 up-regulated 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 de-creased 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 pathogene-sis of S.sclerotiorum,and suggested that the autophagy pathway played important roles in the pathogen-esis of S.sclerotiorum.
关键词
核盘菌/SsATG5/SsATG8/自噬/致病力/侵染垫Key words
Sclerotinia sclerotiorum/SsATG5/SsATG8/autophagy/virulence/infection cushion引用本文复制引用
基金项目
国家自然科学基金(32172368)
中央高校基本科研业务费专项(2021ZKPY014)
现代农业产业技术体系建设项目(CARS-12)
出版年
2024
NETL
NSTL
万方数据