双重PCR法同步检测菜豆晕疫病菌和萎蔫病菌  

Duplex PCR detection of Pseudomonas savastanoi pv. phaseolicola and Curtobacterium flaccumfaciens pv. flaccumfaciens in soybean

作　　者：杨万风[1] 刘艳[2] 刘翔[1] 邵沛泽[1] 谌运清[1] 赵文军[3] 

机构地区：[1]连云港出入境检验检疫局,江苏连云港222042 [2]连云港市农业科学院水稻研究室,江苏连云港222001 [3]中国检验检疫科学研究院植物检疫研究所,北京100029

出　　处：《浙江农业学报》2015年第9期1612-1618,共7页Acta Agriculturae Zhejiangensis

基　　金：国家质检总局科技计划项目(2013IK277)

摘　　要：为建立同步检测菜豆晕疫病菌和细菌性萎蔫病菌的双重PCR方法,根据Gen Bank上公布的菜豆晕疫病菌arg K基因特异性序列设计1对特异性引物PSPF1/PSPR2,将设计的引物与已发表的检测菜豆细菌性萎蔫病菌特异性引物Cff F1/CffR2结合,经过条件优化,建立了双重PCR反应体系,并进行特异性和灵敏度验证。结果显示,采用双重PCR体系,能从菜豆晕疫病菌和细菌性萎蔫病菌基因组DNA以及人工模拟带菌大豆种子样品中扩增到特异性条带,表明本研究所建立的双重PCR检测方法能同时检测出菜豆晕疫病菌和细菌性萎蔫病菌,可用于口岸进口大豆中2种检疫性细菌的检测。The objective of this study was to establish a duplex PCR method for simultaneous detection of Pseudomonas savastanoi pv. phaseolicola（ Psp） and Curtobacterium flaccumfaciens pv. flaccumfaciens（ Cff）. Based on the arg K gene of Psp in Gen Bank,the primers PSPF1 / PSPR2 were designed. The duplex PCR assay was developed using the combining primers PSPF1 / PSPR2 and Cff F1 / CffR2,which were specific primers for Cff gene. The reaction conditions were optimized and the specificity and sensitivity of the duplex PCR were tested. The expected DNA fragment was specifically amplified from the genomic DNA of Psp and Cff. Specificity was confirmed in the artificially inoculated soybean samples imported. Thus,the duplex PCR developed in this study could be used for the simultaneous detection of the two pathogens from imported soybean.

关 键 词：菜豆晕疫病菌 菜豆细菌性萎蔫病菌 双重PCR 检测方法 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]