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将具有典型葡萄卷叶病(Grapevine leafroll diseas,GLRD)症状的葡萄组织,经差速和硫酸铯—蔗糖密度梯度离心,提纯了GLRV,并制备了兔抗血清。电镜下可观察到长度从600～2000nm的线形病毒颗粒,其中以1400nm左右为主。免疫电镜结果表明线形病毒颗粒能被美国的NY-1分离株抗血清(Ⅲ型)所修饰。在间接ELISA中提纯制品与GLRV的Ⅲ、Ⅳ、Ⅱ型抗血清均能产生免疫反应。与Ⅲ型抗血清产生较强的免疫反应,Ⅳ型次之,Ⅱ型最弱。在SDS-免疫双扩散实验中病组织韧皮部粗提液与GLRV的Ⅲ,Ⅳ、Ⅱ型抗血清均产生免疫沉淀线。从而推测我国葡萄园内的葡萄卷叶病很可能由2种或3种卷叶病毒感染所致.采用A蛋白夹心酶联免疫吸附试验(PAS-ELISA)检测葡萄试管苗,Ⅲ型抗血清和自制抗血清的平行测试结果基本相符,共获得11个生食葡萄和10个山葡萄品种的脱葡萄卷叶病毒和扇叶病毒的组培苗,扩繁后田间试种表现出良好的农艺性状。
GLRV was purified from grape tissue with typical symptoms of Grapevine leafroll diseas (GLRD) by differential centrifugation and cesium-sucrose density gradient, and rabbit antiserum was prepared. Electron microscopy can be seen from the length of 600 ~ 2000nm linear virus particles, of which about 1400nm dominated. Immunoelectron microscopy results showed that the linear virus particles could be modified by NY-1 antiserum (type Ⅲ) in the United States. In indirect ELISA purified products and GLRV Ⅲ, Ⅳ, Ⅱ type antiserum can produce immune response. And type Ⅲ antiserum produced a strong immune response, followed by type Ⅳ, type Ⅱ weakest. SDS-immunodiffusion double-diffusion experiments in the diseased phloem crude extract and GLRV type Ⅲ, Ⅳ, Ⅱ antiserum produced immunoprecipitation line. Therefore it is speculated that grapevine leaf curl in our vineyard may be caused by two or three kinds of leafroll virus infection.Plasma ELISA and PAS-ELISA were used to detect grapevine in vitro, type Ⅲ antiserum and The parallel test results of homemade antiserum were basically consistent, and a total of 11 green grapes and 10 grapevine grapevine leaf grafts were obtained. The field experiments showed good agronomic traits.