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OA
Sequence Analysis and Prokaryotic Expression of Pathogenic Factor pglA Gene Derived from the Sugarcane Ratoon Stunting Disease Pathogen
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 Lxx CTCB07 and Lxc DSM46306 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.
梁永检;宋修鹏;颜梅新;王泽平;黄冬梅;黎海涛;韦振飞;张小秋
广西南亚热带农业科学院研究所,广西崇左 532415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广西大新湘桂制糖有限公司,广西崇左 532300横州市植物保护站,广西南宁 530300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
植物保护学
甘蔗宿根矮化病pglA基因克隆
Sugarcaneratoon stunting diseasepglA genecloning
《农业研究与应用》 2023 (5)
1-7,7
广西自然科学基金项目(2020GXNSFBA2970402023GXNSFAA026495)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).
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