绿色荧光蛋白(GFP)标记杧果细菌性角斑病病原菌及其定殖规律  

Colonization rule of GFP-tagged Xanthomonas citri pv.mangiferaeindicae in different tissues of mango

作　　者：於浩然 吴婧波[1] 詹儒林 柳凤[1] 姚全胜[1] 李国平[1] 魏卿[3,4] YU Haoran;WU Jingbo;ZHAN Rulin;LIU Feng;YAO Quansheng;LI Guoping;WEI Qing(South Subtropical Crops Research Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Hainan Province for Postharvest Physiology and Technology of Tropical Horticultural Products/Key Laboratory of Tropical Fruit Biology,Ministry of Agriculture&Rural Affairs,Zhanjiang,Guangdong,524091,China;Yunnan Agricultural University,Kunming,Yunnan,665099,China;Haikou Experimental Station,Chinese Academy of Tropical Agricultural Sciences,Haikou,Hainan,571101,China;Sanya Research Institute,Chinese Academy of Tropical Agriculture Sciences Sanya,Hainan,572025,China)

机构地区：[1]中国热带农业科学院南亚热带作物研究所/海南省热带园艺产品采后生理与保鲜重点实验室/农业部热带果树生物学重点实验,广东湛江524091 [2]云南农业大学,昆明665099 [3]中国热带农业科学院海口实验站,海口571101 [4]中国热带农业科学院三亚研究院,海南三亚572025

出　　处：《中国南方果树》2024年第4期71-79,共9页South China Fruits

基　　金：云南省科技厅科技计划(202104BI090012);中国热带农业科学院基本科研业务费专项资金(1630062022014)资助。

摘　　要：为研究杧果细菌性角斑病病菌在杧果不同器官组织中的定殖规律。利用电击转化法将质粒pBBR1MCS2-Tac-EGFP导入野油菜黄单胞菌杧果致病变种(Xanthomonas citri pv.mangiferaeindicae)菌株Xcm003中,通过转化子生长表型和外源基因的PCR鉴定,成功获得带绿色荧光蛋白(GFP)标记的转化子Xcm003-EGFP,采用室内刺伤和盆栽苗灌根接种试验,结合组织学观察法,追踪该病原菌在不同杧果器官组织中的定殖规律。结果表明,病原菌通过伤口侵入杧果果实果皮外层细胞层,随着侵染时间推移,向内层果皮细胞层垂直扩散并定殖,后期病原菌在伤口处大量聚集,出现隆起开裂状病斑。在杧果叶片中,病原菌从伤口侵入叶片上表皮细胞层,随后迁移到叶肉细胞间隙,侵染后期,病原菌在气孔和伤口定殖,造成气孔和伤口堵塞,导致接种点出现黑褐色水浸病斑。盆栽苗灌根后发现,该病原菌可在土壤中定殖,入侵杧果根部,但并不扩散至其他杧果组织,不会为害整株幼苗。说明杧果细菌性角斑病病菌可在不同杧果组织中定殖,但仅表现为局部侵染特性。To explore the colonization of Xanthomonas citri pv.mangiferaeindicae strains in different tissues of mango,plasmid pBBR1MCS2-Tac-EGFP carrying GFP gene was successfully transformed into the pathogen strain Xcm003 by using electro-transformation method.The GFP labeled transformant Xcm003-EGFP was successfully obtained by identification of growth phenotype and exogenous gene of transformant.The colonization rule of this pathogen in different tissues of mango was tracked using indoor puncture and root irrigation inoculation test combined with histological observation method.The results showed that the pathogen firstly invaded the epicarp cell layer of mango fruit through the wound,and then spread vertically to the endocarp cell layer and colonized with the passage of time.In the late stage of inoculation,the pathogen multiplied in the wound and the bulge cracking spots appeared.In the leaves,the pathogen invaded the epidermis cell layer of mango leaf from the wound,then moved to the mesophyll cell space,and eventually colonized in the stomata and wounds and blocked the stomata and wounds,resulting black-brown‘water stain’disease speckle appearing at the inoculation site.Root-irrigation experiments showed that the pathogen could colonize in soil and invade the root of mango,but not spread to other mango tissues.The results showed that the pathogen could colonize in different mango tissues,but it only showed characteristics of local infection.

关 键 词：杧果细菌性角斑病 野油菜黄单胞菌杧果致病变种Xanthomonas citri pv.mangiferaeindicae GFP标记 质粒 定殖 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]