桃潜隐花叶类病毒RT-PCR和分子杂交检测技术的建立  

The estalishment of RT-PCR and molecular hybridization technique for the detection of peach laten mosaic viroid.

作　　者：徐文兴[1] 王国平[1] 洪霓[1] 

机构地区：[1]华中农业大学植物科技学院,武汉430070

出　　处：《植物检疫》2005年第5期268-271,共4页Plant Quarantine

基　　金：科技部863课题(项目编号:2001AA241142)

摘　　要：本研究从武汉周边采集表现典型花叶症状的桃样品,提取总RNA为模板,采用RT-PCR方法对这些样品进了分析,结果显示所分析的5个样品(P1～P5)均获得了预期大小约为337bp的目标扩增条带,表明样品均带有PLMVd.通过回收RT-PCR产物,用生物素标记制备探针,分别采用DNA斑点杂交、RNA斑点杂交和组织印迹杂交3种杂交方法对这些样品进行检测比较,3种杂交方法中,组织印迹杂交操作步骤相对简单快捷,适合于对PLMVd进行大田快速检测.The peach samples showing typical mosaic were collected from Wuhan area. RT-PCR was employed for the detection of PLMVd by using total RNA as template. Results showed a PCR fragment （337bp） was amplifide from five samples, which indicated that these samples were infected by PLMVd. RT-PCR products was collected and labeled with biotin for probe preparation. Five samples were tested by three molecular hybridization ways including DNA dot blot, RNA dot blot and tissue printing blot. The detection results in dicated that the prepared probe had a good specificity. Among three hybridization methods, tissue printing blot is relativdy simple, rapid. This method is useful for detecting PLMVd in large scale samples.

关 键 词：桃潜隐花叶类病毒 RT-PCR 分子杂交 

分 类 号：S41-30[农业科学—植物保护]