应用锁式探针技术检测大豆细菌性斑疹病  

Development of Padlock Probe Based Method for Detection of Bacterial Pustule of Soybean(Glycine max)

作　　者：陈艳鸿[1,2] 田艳丽[1,2] 赵玉强 胡白石[1,2,4] 

机构地区：[1]南京农业大学植物保护学院,南京210095 [2]农作物生物灾害综合治理教育部重点实验室,南京210095 [3]上海农业技术推广服务中心,上海201103 [4]国家瓜类工程技术研究中心,昌吉831100

出　　处：《农业生物技术学报》2015年第10期1377-1385,共9页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展计划(863)项目(No.2012AA101501);上海市农业技术推广服务中心科技发展基金项目(No.3031344)

摘　　要：大豆(Glycine max)细菌性斑疹病是危害大豆的重要种传细菌病害。近年在全国发病逐年加重,生产无病种子是控制该病害的关键。本研究以看家基因DNA复制和修复蛋白基因(DNA replication and repair protein,recF)为靶标,建立基于锁式探针(Padlock)结合正向斑点杂交技术的地毯草黄单胞菌大豆致病变种(Xanthomonas axonopodis pv.glycines)检测体系。结果表明,该锁式探针能够特异性识别大豆细菌性斑疹病菌,检测灵敏度达到100 fg/μL。该系统对人工模拟种子带菌检出率达到0.1%,利用建立的检测体系对市售大豆种子进行了实际检测应用,从市售的45份大豆种子中检出4份带有大豆细菌性斑疹病菌。本研究表明,基于锁式探针结合正向斑点杂交技术的检测体系能够准确识别大豆细菌性斑疹病菌,从而为该病害的预防和控制提供了有益的参考。Bacterial pustule is a serious seed-borne disease,which dramatically reduces soybean（Glycine max） yield.The disease becomes more and more serious around the country these years.This research selected house-keeping gene of DNA replication and repair protein（rec F） as target gene,and developed a bacterial pustule detection system based on padlock probe combined with dot-blot hybridization.The results indicated that the Padlock probe could detect the bacterial pustule pathogen specifically,and its sensitivity could be up to 100 fg/μL.The system also facilitated the detection of Xanthomonas axonopodis pv.glycines in soybean seedlots with 0.1% artificially infested seed.On the other hand,45 samples of commercial soybean seeds were detected by using this method,and detected 4 samples positive with the bacterial pustule pathogen.The results indicated that the detecting system based on padlock probe combined with dot-blot hybridization could detect Xanthomonas axonopodis pv.glycines accurately,and provide useful references for prevention and control of bacterial pustule.

关 键 词：锁式探针 大豆细菌性斑疹病 检测 DNA复制和修复蛋白基因(recF) 

分 类 号：S565.1[农业科学—作物学]