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.
出处
《植物病理学报》
CAS
CSCD
北大核心
2005年第5期420-427,共8页
Acta Phytopathologica Sinica
基金
K035-2000,supportedbyGeneralAdministrationofQualitySupervision,InspectionandQuarantineofthePeoplesRepublicofChina
