甘蔗宿根矮化病病原菌致病因子pglA基因的序列分析与原核表达  

Sequence Analysis and Prokaryotic Expression of Pathogenic Factor pglA Gene Derived from the Sugarcane Ratoon Stunting Disease Pathogen

作　　者：梁永检 宋修鹏[1] 颜梅新[1] 王泽平[1] 黄冬梅 黎海涛 韦振飞 张小秋 LIANG Yongjian;SONG Xiupeng;YAN Meixin;WANG Zeping;HUANG Dongmei;LI Haitao;WEI Zhenfei;ZHANG Xiaoqiu(Sugarcane Research Institute,Guangxi Academy of Agricultural Sciences/Guangxi Key Laboratory of Sugarcane Genetic Improvement/Key Laboratory of Sugarcane Biotechnology and Genetic Improvement(Guangxi),Ministry of Agriculture and Rural Affairs,Nanning,Guangxi 530007,China;Guangxi South Subtropical Agricultural Science Research Institute,Longzhou,Guangxi 532415,China;Guangxi Daxin Xianggui Sugar Co.,Ltd.,Chongzuo,Guangxi 532300,China;Hengzhou Plant Protection Station,Hengzhou,Guangxi 530300,China)

机构地区：[1]广西农业科学院甘蔗研究所/广西甘蔗遗传改良重点实验室/农业农村部广西甘蔗生物技术与遗传改良重点实验室,广西南宁530007 [2]广西南亚热带农业科学院研究所,广西崇左532415 [3]广西大新湘桂制糖有限公司,广西崇左532300 [4]横州市植物保护站,广西南宁530300

出　　处：《农业研究与应用》2023年第5期1-7,共7页Agricultural Research and Application

基　　金：广西自然科学基金项目(2020GXNSFBA297040,2023GXNSFAA026495);国家自然科学基金青年科学基金项目(32001606);广西科技重大专项(桂科AA22117004,桂科AA22117002);广西创新团队建设项目(nycytxgxcxtd-2021-03-06);广西农业科学院基本科研业务专项(桂农科2021YT007)。

摘　　要：多聚半乳糖醛酸内切酶(endo-PG)是植物与微生物互作中极其重要的细胞壁降解酶,是病原菌定殖寄主与产生致病性的必需因子,革兰氏阴性菌Leifsonia xyli subsp. xyli (Lxx)是甘蔗宿根矮化病的致病菌,pglA是Lxx的关键致病因子。为了揭示pglA基因的功能,本研究采用重叠延伸PCR的方法获得了pglA基因的开放阅读框(ORF),ORF长1491 bp的,编码496个氨基酸,序列保守结构域包含Gly_hydro_28 (Polygalacturonase)与Rho超家族,与Lxx CTCB07、Lxc DSM46306归为一类,相似性达100%。氨基酸序列分析显示,pglA蛋白属于疏水性蛋白,包含信号肽和跨膜区。在大肠杆菌中表达纯化获得pglA蛋白,以包涵体的形式存在于菌体中。研究结果为进一步进行Lxx的致病机制研究奠定了基础。Endo-polygalacturonase(endo-PG),a crucial cell wall-degrading enzyme for plant-microorganism interaction,was an essential factor for colonization and pathogenicity of phytopatho‐gen.Leifsonia xyli subsp.xyli(Lxx)is the pathogen of ratoon stunting disease(RSD)and pglA is the key pathogenic factor of Lxx.In order to reveal the function of pglA,through method of splice over‐lap extension(SOE)PCR,a 1491 bp-long opening reading frame(ORF)encoding 496 amino acids,conserved domains of Gly_hydro_28(polygalacturonase)and Rho superfamily were found con‐tained in pglA gene and the gene was clustered with LxxCTCB07 and LxcDSM46306 in the same group,presenting a 100%similarity between the two strains.The analysis of amino acid sequence showed that pglA protein was hydrophobic containing the signal peptides and transmembrane re‐gions.pglA protein was expressed in Escherichia coli after purification,existing in the thallus in the form of inclusion body.The study results lays a foundation for further research on Lxx pathogenic mechanism.

关 键 词：甘蔗 宿根矮化病 pglA基因 克隆 

分 类 号：S432.1[农业科学—植物病理学]