葡萄扇叶病毒的分离、鉴定、提纯及血清学研究  

ISOLATION, IDENTIFICATION, PURIFICATION AND SEROLOGY OF GRAPEVINE FAN LEAF VIRUS

作　　者：蔡文启[1] 郭德银[2] 徐绍华[1] 王荣[1] 莽克强[1] 

机构地区：[1]中国科学院微生物研究所 [2]中国农业科学院郑州果树研究所

出　　处：《植物病理学报》1990年第2期99-105,共7页Acta Phytopathologica Sinica

基　　金：国家料委<七五>攻关项目

摘　　要：从表现扇叶和叶肉褪绿斑驳症状的先锋、葡萄园皇后和北醇的杂交单株中分离到3个与国外扇叶病毒(GFV)DIL分离物相似的GFY-DIL类似物。它们在昆诺藜、苋色藜、千日红和菜豆上表现症状。提纯病毒颗粒为直径约30nm的球形颗粒。在免疫双扩散试验中均与扇叶病毒抗血清形成完全融合的沉淀线。免疫电镜下病毒颗粒受扇叶病毒抗血清的修饰。利用三种间接异种动物抗体双夹心法和金黄色葡萄球菌蛋白A夹心酶联免疫吸附试验(PAS—ELISA)从感病的葡萄植株或组培苗叶片中检测了GF。间接异种动物抗休双夹心法能检测提纯GFV的最低浓度为1.6ng—8ng/ml。春季的芽和幼叶为合适的检测器官。Three GFV DIL-like were obtained by mechanical inoculation to Cheno podium quinoa from vines of Vitis vinifera cv.Xianfeng and progenies of cv. the Qreen of Vineyard and cv. Beichun.The symptoms were similar to those caused by grapevine fan leaf virus (GFV) on C, quinoa, C. amaranticolor, Gomphrena globosa and phaseolus vulgaris cv. Bountiful.In buffered sap of C. quinoa,their dilution end point (DEP) ranged from 1: 2000 to 1: 5000, thermal inactivation point(TIP)was between 59-62℃ and longevity invitro was more than 4 weeks at room temperature in spring. The virus was propagated on C. quinoa and purified by chloroform-butanol clarification, PEG_(6000)precipitation and ultracen trifugation. It resulted in a higher production c, 1.8-3,5mg virus per 100g infected tissues. Amax=200nm Amin=240nm A200/A280=1.72. The particles are isometrically spherical c,30nm in diameter. Some particles appear hollow. In gel diffusion test the three GFV-DIL-like reacted against GFY-DIL antiserum and also decorated by the antiserum against GFV in immunosorbent electronmicroscopy(ISEM).They did not react with antisera of ArMV,GBLV, GYVV,TRSV and GCMV. As a result, the three DIL-like were identified as GFV. We have detected fanleaf virus from different grapevine organs and plantlets by PAS-ELISA and indirect DAS-ELISA using mouse polyclonal ascites fluid, rabbit antiserum and immunoglobulin of chicken. The sensitivity of indirect DAS-ELISA to detect purified virus is I.0-8ng/ml, while in the infectivity test the minimum concentration of the purified virus used is 1,000ng-500,000ng/ml. The best source of antigen for indirect ELISA tests appeared to be bud and young leaf tissues in spring. Nicotine containing extraction medium or Tris-HCl 0.5M pH 8.2,2% PVP, 0.8% NaCl are suitable for detecting.

关 键 词：葡萄 扇叶病毒 分离 提纯 血清学 

分 类 号：S436.631.1[农业科学—农业昆虫与害虫防治]