树生黄单胞菌李致病变种两种PCR检测方法的建立  

Two specific PCR assays for detection of Xanthomonas arboricola pv.pruni

作　　者：杨雯迪 马玉超 王金鹏 沈建国 任争光[1] 杨海清 李为民 YANG Wendi;MA Yuchao;WANG Jinpeng;SHEN Jianguo;REN Zhengguang;YANG Haiqing;LI Weimin(Department of Plant Protection,Beijing University of Agriculture,Key Laboratory for Northern Urban Agriculture,Ministry of Agriculture and Rural Affairs,Beijing 102206,China;Fujian Key Laboratory for Technology Research of Inspection and Quarantine,Technology Center of Fuzhou Customs District,Fuzhou 350001,China;Fruit Industry Service Center,Pinggu Bureau of Agriculture and Rural Affairs,Beijing 101200,China)

机构地区：[1]北京农学院植物保护系,农业农村部华北都市农业重点实验室,北京102206 [2]福建省检验检疫技术研究重点实验室,福州海关技术中心,福州350001 [3]平谷区果品产业服务中心,北京101200

出　　处：《植物保护》2024年第5期224-229,237,共7页Plant Protection

基　　金：国家自然科学基金(32170190)。

摘　　要：树生黄单胞菌李致病变种Xanthomonas arboricola pv.pruni(Xap)是桃、李生产中最具毁灭性的病原细菌之一。为了快速准确地检测Xap,本研究以其编码基因L1B 16_06350的5′端序列(611 bp)为检测靶标,开发了常规PCR和TaqMan实时PCR检测方法。这两种方法与树生黄单胞菌胡桃致病变种X.arboricola pv.juglandis等8个不同属的11种细菌均没有交叉反应,其中常规PCR检测极限为10^(-2)ng,TaqMan探针法为10^(-3)ng,表现高度的灵敏度和特异性。利用田间样品进行测试,结果证实两种检测方法均可应用于Xap的常规检测。Xanthomonas arboricola pv.pruni(Xap)is one of the most devastating bacterial pathogen to peach and plum.To rapidly and accurately detect Xap,we developed a conventional PCR and a TaqMan real-time PCR assay that targets the 5′end(611 bp)of L1B 16_06350,a Xap encoding gene.Both methods had no cross-reaction with 11 tested bacterial species from eight different genera,including X.arboricola pv.juglandis etc.,with a limit of detection of 10^(-2)ng for the conventional PCR and 10^(-3)ng for the TaqMan method,respectively,showing high sensitivity and specificity.Field sample test proved the reliability of the two methods in routine diagnosis of Xap.

关 键 词：树生黄单胞菌李致病变种 常规PCR TaqMan探针法 特异性 灵敏度 

分 类 号：S436.62[农业科学—农业昆虫与害虫防治]