南方水稻黑条矮缩病毒快速检测.pdfVIP

基因组学与应用生物学，2010 年，第29 卷，第5 期，第1009-1012 页 Genomics and Applied Biology, 2010, Vol.29, No.5, 1009-1012 技术改进 Upgraded Technology 南方水稻黑条矮缩病毒快速检测 1,2 1 1 1 1,2* 周倩 朱俊子 梁晋刚 陈欣怡 高必达 1 湖南农业大学生物安全科学技术学院, 长沙, 410128; 2 植物疾病控制与利用湖南省高校重点实验室, 长沙, 410128 * 通讯作者, bdgao@ 摘 要 南方水稻黑条矮缩病毒(southern rice black-streaked dwarf virus, SRBSDV) 引起的新水稻矮缩病近 年在越南北部和我国南方各省爆发，准确快速地检测病毒是病害预测的关键。本文针对该病毒的S10 序列 设计了一对新的引物S10F/S10R，利用一步法RT-PCR ，对2010 年5~8 月间湖南省下属各县送来的疑似感 染该病毒引起的水稻矮缩病样本进行了检测，结果显示，新引物能有效地区分阳性样品和非阳性样品。同时 对PCR 产物进行了序列测定，结果表明序列均与已发表的南方水稻黑条矮缩病毒序列(登录号: EU523360 和 EU784840)达约99%的同源性。还在此基础上构建了系统发育树，发现SRBSDV 湖南、广东和海南分离物病毒 位于一个相对独立的分支。研究发现相对以往的巢式RT-PCR，本文采用的新引物及一步法RT-PCR 能快速检 测南方水稻黑条矮缩病毒，简化了操作步骤，缩短了检测时间，适合在SRBSDV 检测和病害预测中应用。 关键词 南方水稻黑条矮缩病毒, RT-PCR, 系统发育树, 快速检测 Rapid Detection of Southern Rice Black-Streaked Dwarf Virus Zhou Qian 1,2 Zhu Junzi 1 Liang Jingang 1 Chen Xinyi 1 Gao Bida 1,2* 1 College of Bio-Safety Science and Technology, Hunan Agricultural University, Changsha, 410128; 2 Hunan provicial university key lab for plant dis- ease control and ulitization, Changsha, 410128 * Corresponding author, bdgao@ DOI: 10.3969/gab.029.001009 Abstract Rice dwarf caused by southern rice black-streaked dwarf virus (SRBSDV) was outbreak in the North- ern of Vietnam and many provinces in Southern China in recent years. So, rapid and accurate detection of the virus is the key to forecaste the disease. In this paper, we designed a new pair primer S10F/S10R based on SRBSDV S10 sequence. And then we detected the rice dwarf samples, which suspect infected this virus, from different counties of Hunan Province between May to July 2010 by one step RT-PCR. The result showed that the new primer could effectively distinguish the positive and negative samples. At the same time, the PCR products were sequenced, the result demonstrated that both the sequences had the homology 99% comparing with the published sequence of southe