洋葱腐烂病菌PCR检测方法的建立  

Development of PCR detection for Burkholderia gladioli pv.allicola

作　　者：丁钿 魏霜 章柱 李献锋 陈青 林晓红 刘琼光[4] Ding Tian;Wei Shuang;Zhang Zhu;Li Xianfeng;Chen Qing;Lin Xiaohong;Liu Qiongguang(Guangzhou Baiyun Airport Customs House Comprehensive Technical Service Center,Guangzhou 510400,China;Guangzhou Customs District Technology Center;Xiamen Customs District Technology Center;College of Plant Protection,South China Agricultural University)

机构地区：[1]广州白云机场海关综合技术服务中心,广东广州510400 [2]广州海关技术中心 [3]厦门海关技术中心 [4]华南农业大学植物保护学院

出　　处：《植物检疫》2023年第6期24-30,共7页Plant Quarantine

基　　金：国家重点研发计划(2022YFC2601500)。

摘　　要：为了快速准确检测洋葱腐烂病菌(Burkholderia gladioli pv. alliicola,简称Bga),根据GenBank中Bga与相关种的序列差异设计特异引物BG5/BG7和探针Bga-P,建立了Bga常规PCR和荧光PCR检测方法。测试结果表明,引物和探针对供试的11株Bga菌株表现为阳性反应,其他64株供试菌株和空白对照均为阴性。常规PCR和荧光PCR方法的检测灵敏度分别为24 pg和240 fg菌体DNA。美国、法国、意大利等国进境的80批次洋葱种子样品的检测结果显示,这两种方法的阳性检出率分别为7.5%和12.5%。选取阳性样品进行病菌分离,成功从2批次法国进境洋葱种子中分离到目的菌。本文建立的洋葱腐烂病菌PCR检测方法可为口岸检测部门提供更高效、灵敏和特异的检测手段。In order to rapidly and accurately detect Burkholderia gladioli pv.allicola(Bga),the specific primer BG5/BG7 and probe Bga-P were designed according to the sequence difference between Bga and related strains in GenBank.The detection methods of Bga by conventional PCR and fluorescent PCR were established.The conventional PCR with primer BG5/BG7 and the real time PCR with probe Bga-P could amplify 11 strains of Bga,but no target band or positive amplification for other 64 strains of relat-ed species.These two methods could detect as low as 24 pg and 240 fg cell DNA of Bga,respectively.80 batches of onion seed samples imported from the United States,France and Italy showed positive de-tection rates of 7.5%and 12.5%for these two methods,respectively.Positive samples were selected for pathogen isolation,and the Bga were successfully isolated from 2 batches of onion seeds imported from France.These PCR detection method for Burkholderia gladioli pv.allicola can provide a more efficient,specific and sensitive detection method for port inspection departments.

关 键 词：洋葱腐烂病菌 检测 PCR 

分 类 号：S41[农业科学—植物保护]