侵染波斯毛莨的蚕豆萎蔫病毒的鉴定与提纯

从波斯毛莨花卉上分离到一株病毒分离物R89—1,浸染波斯毛莨叶片表现为不规则的黄斑和坏死斑,易经汁液摩擦接种.可经桃蚜以非持久性方式传播.能侵染测试的11科40种植物中的7科23种.病毒粒体为正20面体,直径约25nm.在琼脂双扩散试验中与蚕豆萎蔫病毒(B—BWV)的抗血清发生沉淀反应.根据上述特征,K89—1分离物鉴定为蚕豆萎蔫病毒.蚕豆萎蔫病毒用感染的昆诺阿藜叶片通过丁醇澄清及聚乙二醇沉淀容易提纯,每100g叶组织可获22mg病毒.提纯的病毒制剂具有典型的核蛋白紫外吸收曲线,最大吸收波长为259nm,最小吸收波长为241nm,A260/A280为1.67.在蔗糖密度梯度离心中含有上、中、下三条沉淀带.利用提纯的病毒制备的抗血清经微量沉淀试验效价达1/2048.

Identification and Purification of Broad Bean Wilt Virus Naturally Infectted on Persian Butterrcup

A virus isolate, R89-1, was obtained from the ornamental plant of persian buttercup (Ranunculus asaiaticus L. ). Unregular yellow spots and necrosis of the leaves were observed when the plants were infected with R89-1. It could be transmitted by sap and aphids (myzus persicae) in a nonpersis-tant manner. 23 of 40 species of tested plants in 7 of 11 families could be infected with R89-1 exper-imently. Many isometric particles, about 25 nm in diameter were observed under electron microscopy. The virus stronly reacted with broad bean wilt virus (BBWV) antiserum not CMV antiserum in agar double diffusion tests. Based on the characteristics mentioned above, R89- 1 was identified as BBWV.the virus, BBWV, was readily purified by n-butyl alcohol-PEG precipitation procedure using infected leaves of Chenopodium quinoa. The yield of the virus was about 22 mg of viruses per 100 g of infected tissues. Purified virus preparations almost contained plant impurities and had an absorption spectrum with a maximum at 259 nm and a minimum at 241 nm typical of a spherical virus. The A260/A280 ratio was 1. 67. When the preparations were centrifuged in sucrose density gradients, three bands, such as top, middle, and bottom, were observed. The antiserum prepared using purified virus preparations had a titre of 1/2048 as determined by microprecipitin tests.