水稻黑条矮缩病毒检测体系的优化及田间寄主检测  

Optimization of RT-PCR detection system for Rice black-streaked dwarf virus and detection of its natural hosts

作　　者：阴筱[1] 朱芹芹[1] 许斐斐[1] 刘志强[1] 张广民[1] 李向东[1] 

机构地区：[1]山东农业大学植物保护学院植物病毒研究室,泰安271018

出　　处：《植物病理学报》2013年第6期647-650,共4页Acta Phytopathologica Sinica

基　　金：山东省科技攻关项目(2009GG10009021);山东省现代农业产业技术体系

摘　　要：玉米粗缩病20世纪50年代和90年代中后期在我国部分地区严重发生，2008年至今，该病在黄淮海地区又呈暴发趋势。To establish and optimize the RT-PCR detection system of Rice black-streaked dwarf virus （ RBS- DV）, and identify the natural host of RBSDV in the field, four pairs of primers were designed according to the nucleotide sequences of RBSDV S10. RT-PCR detection system for RBSDV was optimized after comparing pri- mer pair, concentration of Mg2＋ and Taq polymerase, and times of RNA dilution. The fourth pair of primers （ F4 and R4） was proved to be the most sensitive one. The optimal RT-PCR system was 10x PCR buffer 2.5 μL, 25 mmol/L MgC12 1.5 μL, dNTP （each 2.5 mmol/L） 2.0 μL, primers F4 and R4 （10 μmol/L） each 1.0 μL, 5 U/ μL Taq DNA polymerse 0.2 μL, template RNA 5.0 μL and sterilized water 11.8 μL to make a total volume of 25 μL. With the method established, RBSDV could be detected from RNA extracted from single Laodelphax striatellus or 30 ng infected maize leaves. Besides plants of family Gramineae, Sonchus brachyotus and Eclipta prostrate from family Asteraceae, and Amaranthus retroflexus from Amaranthaceae were also natural hosts of RBSDV.

关 键 词：病毒检测 寄主 田间 优化 黑条 水稻 玉米粗缩病 黄淮海地区 

分 类 号：S476.3[农业科学—农业昆虫与害虫防治]