16S nested-PCR技术检测玉米细菌性枯萎病菌(英文)  

Identification of Corn Bacterial Wilt Pathogen by Nested-PCR Based on 16S rDNA

作　　者：吴琼[1] 陈枝楠[1] 范怀忠[2] 金显忠[1] 

机构地区：[1]深圳出入境检验检疫局,深圳518010 [2]华南农业大学资环学院植物病理学系,广州510640

出　　处：《植物病理学报》2005年第5期420-427,共8页Acta Phytopathologica Sinica

基　　金：K035-2000,supportedbyGeneralAdministrationofQualitySupervision,InspectionandQuarantineofthePeoplesRepublicofChina

摘　　要：玉米细菌性枯萎病是玉米上的重要种传病害,病原菌为Pantoea stewartiisubsp.stewartii。本研究设计了16S通用引物,扩增该病菌及其近似种的16S rDNA,通过序列测定和分析,针对该病菌设计了特异性引物,采用nested-PCR技术,能够准确地区别该病菌及其近似种,检测的灵敏度在DNA水平上达到10-3pg级,检测活菌则达到2 cfu。检测人工污染的玉米种子时,不受种子提取液中其它物质的干扰,灵敏度依然达到2 cfu。Pantoea stewartii subsp, stewartii is the seed-borne causative agent of bacterial wilt and leaf blight of sweet corn and maize. In this study, a specific 16S rDNA-based nested-PCR （nPCR） was developed to identify the bacterium from a range of closely related bacteria. This PCR assay was proved to be highly sensitive with the detection limit of 10^-3 pg genomi＇c DNA or 2 colony forming units （cfu）. Moreover, this PCR assay successfully detected the target bacteria as few as 2 cfu when tested with artificially contaminated seeds, and the sensitivity was not affected by seed compounds.

关 键 词：玉米细菌性枯萎病菌 16S RDNA 检测 巢式PCR 

分 类 号：S435.131.4[农业科学—农业昆虫与害虫防治]