十字花科黑腐病菌Ⅲ型效应物XopXccR1植物亚细胞定位  

Subcellular localization of a type Ⅲ secreted effector XopXccR1 of Xanthomonas campestris pv.campestris in plant cells

作　　者：冯艳茹 高亚君 杭小红[1] 何勇强[1,2] 姜伯乐 Yanru Feng;Yajun Gao;Xiaohong Hang;Yongqiang He;Bole Jiang(College of Life Sciences and Technology,Guangxi University,Nanning 530004,Guangxi Zhuang Autonomous Region,China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Nanning 530004,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西大学生命科学与技术学院,广西南宁530004 [2]亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004

出　　处：《微生物学报》2020年第11期2572-2581,共10页Acta Microbiologica Sinica

基　　金：广西自然科学基金(2014GXNSFFA118005,2018GXNSFAA281147)。

摘　　要：【目的】植物病原细菌通过Ⅲ型分泌系统(type Ⅲ secretion system,T3SS)将Ⅲ型效应物(type Ⅲ secreted effectors,T3SEs)分泌转运到宿主细胞的不同位点上,进而行使不同的致病功能。本研究旨在确定Xcc 8004 Ⅲ型效应物中分子量最大的蛋白XopXccR1在植物中的亚细胞定位。【方法】利用生物信息学方法分析XopXccR1的跨膜信息。通过同源重组方法将XopXccR1全长、N端(1–1220aa)和C端(1221–2030 aa)分别克隆到植物表达载体pCAMBIA-2300-35S::EGFP上,利用根癌农杆菌介导的瞬时表达浸染本生烟,通过激光共聚焦显微镜观察亚细胞定位结果。【结果】XopXccR1全长和N端定位在本生烟细胞膜上,而C端定位在细胞质中。【结论】XopXccR1的N端与C端可能分别存在定位信号,N端信号主导全长蛋白的最终定位。[Objective]Plant pathogenic bacteria can directly deliver the secreted type Ⅲ effectors(T3SEs)into different sites of host cells through the type Ⅲ secretion system(T3SS),thereby exercising various pathogenic functions.The purpose of this study was to determine the subcellular localization of the largest T3SE protein XopXccR1 of Xcc 8004 in plants.[Methods]The bioinformatics methods were used to analyze the putative transmembrane region of XopXccR1.To obtain the transient expression Agrobacterium strains,the full-length XopXccR1 without stop codon,the N-terminus(1–1220 aa),and the C-terminus(1221–2030 aa)were ligated to the plant expression vector pCAMBIA-2300-35S::EGFP respectively by homologous recombination.Utilizing Agrobacterium-mediated transient expression to infect Nicotiana benthamiana,the results of subcellular localization were then observed with a laser confocal microscope.[Results]The full-length and the N-terminus of XopXccR1 are mainly localized in the cell membrane of N.benthamiana,while the C-terminus of XopXccR1 in the cytoplasm.[Conclusion]XopXccR1 N-and C-terminus might both have the targeting signals.The targeting signals in the N-terminus of XopXccR1 mainly contributes the final sub-localization of XopXccR1.

关 键 词：十字花科黑腐病菌 Ⅲ型效应物 XopXccR1 亚细胞定位 

分 类 号：S432.42[农业科学—植物病理学]