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侵染西瓜的黄瓜绿斑驳花叶病毒快速检测

宋西娇，李艳敏，王荣洲，吴宏宇，吴建祥，洪 健*
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2016年 第35卷 第4期: １０００⁃６２８１（２０１６）０４⁃０３５６⁃０５     下载地址：

侵染西瓜的黄瓜绿斑驳花叶病毒快速检测

宋西娇1,3，李艳敏2，王荣洲2，吴宏宇3，吴建祥1，洪 健1*

(1. 浙江大学生物技术研究所，浙江 杭州310058；2. 浙江省植物保护检疫局，浙江 杭州310029;3. 浙江省农业科学院公共平台，浙江 杭州310021）

摘 要  黄瓜绿斑驳花叶病毒（CGMMV）是危害西瓜生产的重要病害，造成西瓜果实腐烂，失去商业价值。应用透射电子显微镜和单抗dot-ELISA技术对采自浙江温岭的西瓜病叶进行快速病原检测，电镜负染色观察到病株汁液中存在大量刚直杆状的病毒粒子，超薄切片观察到细胞质内存在整齐排列的杆状病毒聚集体，与烟草花叶病毒属（Tobamovirus）的形态特征相符；进一步采用dot-ELISA技术进行血清学检测，显示西瓜病叶与CGMMV特异性抗体呈阳性反应，检测结果表明感染西瓜的杆状病毒为CGMMV。从而为侵染西瓜的CGMMV提供快速检测途径。

关键词  西瓜；黄瓜绿斑驳花叶病毒；负染色；超薄切片；dot-ELISA

中图分类号：Q939.4      文献标识码：A    doi:10.3969/j.issn.1000-6281.2016.04.012

 

Fast detection of the watermelon infected Cucumber green mottle mosaicvirus

SONG Xi-jiao1，3，LI Yan-min2，WANG Rong-zhou2，WU Hong-yu3，WU Jian-xiang1，HONG Jian1*

(1. Institute of Bio-technology，College of Agriculture and Biotechnology，Zhejiang University，Hangzhou Zhejiang 310058;2. Bureau of Zhejiang Plant Quarantine，Hangzhou Zhejiang 310029；3. Public Lab，ZhejiangAcademy of Agricultural Sciences，HangzhouZhejiang 310021，China)

Abstract  Cucumber green mottle mosaic virus(CGMMV) is an important viral pathogen, as it infecting watermelon and inducing rotten in fruit which bring about severe economy loss. The transmission electron microscope (TEM) and the dot-ELISA technology based on mono-clonal antibody were used to carry out fast pathogen detection in infected watermelon which collected form Wenling Zhejiang. Under TEM, large amount of Rod-shapedvirus particles were observed in the crude sap of infected water melon leaf by using of the negative stain method. A section of infected leaf revealed that large amount of virus particles in bacilliform were accumulated in cytoplasm which were arranged regularly. From these features obtained from TEM, we judged that the virus in infected watermelon was a member of Tobamovirus. A further detection by dot-ELISA based on the specific mono-clonal antibody against CGMMV was performed to this infected watermelon which results in positive dots. Combining of these results of detection, we concluded that the pathogen of the infected watermelon collected from Wenling Zhejiang is CGMMV.

Keywords  watermelon；Cucumber green mottle mosaic virus；negative stain；ultra-thin section；dot-ELISA

 

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