复合侵染的蚕豆黄花叶病病原诊断  

Diagnosis of the multiple pathogen of broad bean yellow mosaic disease

作　　者：谢礼[1] 刘文洪 洪健[1] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所,浙江杭州310029 [2]浙江中医学院,浙江杭州310053

出　　处：《电子显微学报》2006年第2期168-171,共4页Journal of Chinese Electron Microscopy Society

基　　金：国家自然科学基金资助项目(No.30270873;No.30370305).~~

摘　　要：应用透射电镜、ELISA和RT-PCR技术对浙江丽水的蚕豆黄化花叶病株进行了病原诊断,电镜负染色发现病株汁液中存在线状和球状两种病毒粒子,线状粒子长度在600nm^800nm的占73%,球状粒子直径约26nm;超薄切片观察到细胞质中存在Ⅱ型风轮状内含体及大量电子致密无定型体,还有线粒体增生聚集和膜结构的增生,两类细胞病变现象出现在相邻细胞中;用BBWV 2的单克隆抗体对病汁液ELISA检测结果为阳性反应;用Potyvirus通用引物进行RT-PCR检测有1.7kb特异性扩增片段产生。根据以上结果诊断病原为蚕豆萎焉病毒2(BBWV 2)和菜豆黄花叶病毒(BYMV),二者为复合侵染。The multiple infecting pathogen were detected in a sample of yellow mosaic broad bean from Lishui, Zhejiang province, by transmission electron microscopy, ELISA and RT-PCR. The results showed that two kinds of virus were found in leaf sap negative stained with 2 % PTA. One was filamentous virus particles with 73 % lengths ranging from 600nm to 800nm. The other was spherical virus particles, about 26nm in average diameter. The pathological study revealed that the Ⅱ -type cylindrical inclusions and the amorphous inclusions were appeared in cytoplasm of the infected cells, which is the pathological symptom of BYMV infecting. Mitochondrial aggregation and membranous proliferation were also found in the infected cells, which is the pathological symptom of BBWV 2 infecting. One of the viruses were positive reacted with BBWV 2 monoclonal antibody by ELISA experiment. There was 1.7kb special production in 1% gel electrophoresis after RT-PCR using universal primer of genus Potyvirus. Based on all the results above, suggested that the spherical virus were Broad bean wilt virus 2 （BBWV 2） and the filamentous virus were Bean yellow mosaic virus （BYMV）. They were multiple infecting.

关 键 词：蚕豆 蚕豆萎蔫病毒2 菜豆黄花叶病毒 病原诊断 

分 类 号：S436.43[农业科学—农业昆虫与害虫防治]