PCR技术快速检测玉米内州萎蔫病菌研究  

PCR for rapid identification of Clavibacter michiganensis subsp. nebraskensi

作　　者：周琦[1] 陈寔[2] 高文娜[1,2] 汪万春[1] 李建光[1] 陈洪俊[1] 李健强[2] 

机构地区：[1]北京出入境检验检疫局,北京100026 [2]中国农业大学农业部植物病理学重点开放实验室

出　　处：《植物检疫》2010年第2期16-18,共3页Plant Quarantine

基　　金：国家“十一五”重大科技支撑计划;“农林重大生物灾害防控技术研究”项目;“潜在入侵物种口岸侦测技术”(课题编号:2006BAD08A13);国家质检总局科技计划项目“检疫性植物病原细菌抗体荧光双标记纳米检测技术研究”(课题编号:2009IK251)

摘　　要：玉米内州萎蔫病是北美玉米生产中的严重病害,为防止其传入我国,本研究采用Clavibacter michiganensis不同亚种的转录间隔区序列,设计了特异性的PCR引物,对C.michiganensis种下不同亚种的DNA进行了PCR扩增反应,结果表明,只有玉米内州萎蔫病菌的特异性扩增得到大小为170bp的目标片段,且仅有此一条明显的PCR扩增产物,C.michganensis种下其他亚种无扩增产物。PCR反应的灵敏度为4.36×105cfu/mL和1.56×102pg,可以满足检疫的要求。Goss＇s Bacterial Wilt and Leaf Blight of Corn is a kind of severe disease occurring in the corn production in the North America. To avoid its entry into China, the intergenic transcribed spacer of Clavibacter michiganensis was analyzed to design the PCR primer pairs, which was adopted in the PCR of different C. michiganen- s/s subspecies. The result showed that only one DNA fragment was amplified when the template was C. m. nebraskensis, and no fragment was amplified when other subspecies DNA was used as templates. The sensitivity of the PCR was 4.36 × 10^5 cfu/mL and 1.56 - 102 pg, and this could meet the demand of daily quarantine.

关 键 词：玉米内州萎蔫病菌 PCR 快速检测 

分 类 号：S41-3[农业科学—植物保护]