玉米细菌性枯萎病菌实时荧光PCR检测方法的建立  

Establishment of real-time PCR detection method for Pantoea stewartii subsp.stewartii

作　　者：丁钿 牟桂萍 魏霜 李献锋 林晓红 章柱 易建平 Ding Tian;Mou Guiping;Wei Shuang;Li Xianfeng;Lin Xiaohong;Zhang Zhu;Yi Jianping(Guangzhou Baiyun Airport Customs House,Guangzhou 510400,China;Huangpu Customs District Technology Center;Guangzhou Customs District Technology Center;Technical Center for Animal,Plant and Food Inspection and Quarantine of Shanghai Customs District)

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]黄埔海关技术中心 [3]广州海关技术中心 [4]上海海关动植物与食品检验检疫技术中心

出　　处：《植物检疫》2024年第2期32-37,共6页Plant Quarantine

基　　金：国家重点研发计划(2022YFC2601500)。

摘　　要：玉米细菌性枯萎病菌(Pantoea stewartii subsp. stewartii,Pss)是我国进境植物检疫性有害生物。根据Pss与其近似种序列差异,设计特异性引物PS3F/PS3R和探针PS-3P,建立Pss实时荧光PCR检测方法。测试结果显示,供试的6株Pss菌株均有阳性扩增,其他74株供试菌株和空白对照均为阴性扩增,菌体DNA的检测低限为13 fg;对模拟带菌玉米种子进行实时荧光PCR检测并建立标准曲线,R2=0.998 8,线性关系良好。本文建立的实时荧光PCR检测方法可为口岸检疫提供一种准确快速检测方法。Pantoea stewartii subsp.stewartii(Pss)is one of import plant quarantine pests in china.The specific-Pss primer and probe were designed by comparing the sequence difference between Pss and the related species,and established a real-time fluorescence PCR detection method.The results showed that only 6 strains of Pss had positive amplification,other 74 strains and blank control were negative.The low limit of detection was 13 fg of DNA.The established real-time fluorescence PCR method was used to detect the simulated infected maize seeds and the standard curve was established.It was found that R2=0.9988,showing a good linear relationship.The specific and sensitive PCR method can provide a more accurate and rapid detection method for port quarantine.

关 键 词：玉米细菌性枯萎病菌 实时荧光PCR 检测 

分 类 号：S41[农业科学—植物保护]