应用RT-PCR、斑点杂交法和SDS-PAGE检测水稻黑条矮缩病毒  

Detection of rice black-streaked dwarf Fijivirus by RT-PCR, dot-blot hybridization and SDS-PAGE

作　　者：王朝辉[1] 周益军[2] 范永坚[2] 薛宝娣[1] 吴淑华[2] 程兆榜[2] 张文荟[2] 

机构地区：[1]南京农业大学植保学院,南京210095 [2]江苏省农业科学院植物保护研究所,南京210014

出　　处：《南京农业大学学报》2001年第4期24-28,共5页Journal of Nanjing Agricultural University

基　　金：江苏省九五重点攻关项目 (BY96 386 ) ;江苏省应用基础项目 (BJ97132 )

摘　　要：用RT PCR技术、PCR标记的探针点杂交和SDS PAGE检测了生产上严重危害玉米和水稻的水稻黑条矮缩病毒(RBSDV)。由RT PCR扩增的RBSDV第 7片段第 92 1～ 14 11碱基作探针 ,用PCR法DIG标记后点杂交 ,可以从 10 0ng玉米病叶中检测到RBSDV ,灵敏度是RT PCR的 1/10 ;10 %SDS PAGE只能检测到从 1g玉米病叶中提取的病毒dsRNA ,但对江苏玉米田间分离的不同的RBSDV样品电泳发现 ,该病毒基因组dsRNA有差异 。The purpose of this study was evaluation of RT PCR technique,dot blot hybridization with PCR generated probe and SDS PAGE for the detection of rice black streaked dwarf Fijivirus(RBSDV).The cDNA probe labeled with digoxigenin by PCR,complementary to the region between 921 and 1 411 nucleotides of segment 7 of RBSDV from Jiangsu,was used.In dot blot hybridization the probe could detect the viral RNA from 100 ng diseased maize leaves,which is ten times less sensitive than RT PCR.10% SDS PAGE only could detect the viral dsRNA extracted from 1 g diseased maize leaves,but this technique was found to be simple and effective to study the genomic diversity of RBSDV isolated in fields.

关 键 词：水稻 黑条矮缩病毒 检测 RT-PCR 斑点杂交 SDS-PAGE 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治]