甜菜花叶病毒病的研究Ⅰ.病毒的分离鉴定

 1986-1987年对黑龙江、内蒙古部分甜菜原料产区与采种区进行了初步调查,采种区甜菜花叶病毒病发病率达100%,与其邻近的原料产区有零星发病,远离采种区则没有发病的迹象。田间采集的病叶,用磨擦、蚜传接种方法均能在甜菜上再现症状。经昆诺藜(Chenopodium quinoa)分离,普通电镜及乳胶电镜负染均能看到典型的马铃薯Y组病毒的粒体。通过鉴别寄主、甜菜上的症状表现,分离物的物理特性及血清学试验,证明此分离物为甜菜花叶病毒。在提纯方面,摸索了不同方法,认为匀浆前液氮或冷冻处理,二次PEG沉淀后过Sepharose4B柱,分离病毒效果较好,还可省去超速离心步骤。以内蒙五号甜菜品种作为毒源的繁殖寄主,能使提纯病毒产量达0.6mg/100g叶片,耳静脉与多点皮下注射相结合免疫兔子,微量沉淀测定效价为1:1024。

THE STUDIES OF BEET DISEASE INDUCED BY BEET MOSAIC VIRUS Ⅰ. Isolation and Identification of The Virus

In China, the growth of sugar beet is distributed mainly in Heilongjiang, Xinjiang,and Inner Mongolia。According to surveys made in 1986-1987, the mosaic disease of sugar beet attracted the writer s'attention. The mosaic symptoms were very distinct in fields. Samples collected from fields and inoculated beet seedlings as well as Chenopodium quinoa and other assay plants either by aphids or by sap inoculation demonstrated that this mosaic disesed samples were transmissible.A series of the isolated virus by symptom expression on assay host plants and physical characters so far reported revealed that it is identical to BtMV. Under electron microscopic studies, the typical flexous filamentous particles measured 730×13nm were clearly shown. The sugar beet CV.:Inner Mongolia No.5 is an ideal host for propagation of BtMV. To obtain more pure preparations of the virus, the purification procedures were slightly modified that the infected leaves was processed first by liquid nitrogen or refrigeration before homogenization. After second PEG precipitation, it was passed through a column containing Sepharose 4B. The yield of purified virus was up to 0.6mg/100g of infected leaves and the titre of the antiserum against BtMV was 1:1024 by droplet preciptation tests.