杏鲍菇枯萎病菌PCR检测方法的建立与初步应用  

Establishment and Preliminary Application of PCR Detection Methods for Pantoea pleuroti

作　　者：秦文韬 谷彤彤 刘宇[1] 宋忠娟 荣成博 QIN Wentao;GU Tongtong;LIU Yu;SONG Zhongjuan;RONG Chengbo(Institute of Plant and Environment Protection,Beiing Academy of Agriculture and Forestry Sciences/Beiing Engineering Research Center for Edible Mushroom/Key Laboratory of Urban Agriculture(North China)by Ministry of Agriculture and Rural Affairs,Beiing 100097)

机构地区：[1]北京市农林科学院植物保护环境保护研究所,北京市食用菌工程技术研究中心,农业农村部华北都市农业重点实验室,北京100097

出　　处：《北方园艺》2021年第7期123-128,共6页Northern Horticulture

基　　金：国家自然科学基金青年科学基金资助项目(31701975);北京市农林科学院创新能力建设专项资助项目(KJCX20200105)。

摘　　要：以杏鲍菇枯萎病菌侧耳泛菌(Pantoea pleuroti)为试材,采用PCR引物设计的方法,通过分析P.pleuroti基因组的测序结果,设计合成并筛选出可检测P.pleuroti的引物,并研究其检测效果,以期建立P.pleuroti的高效PCR检测体系,并为科学防控杏鲍菇枯萎病提供分子基础。结果表明:K3143F/R和K37F/R 2对引物特异性强,仅P.pleuroti基因组DNA作为模板时,PCR扩增产物分别呈现1条660 bp和1条666 bp的特异性条带,15株Pantoea属细菌和3株食用菌常见病原菌的基因组DNA及阴性对照作为模板扩增产物均无条带;基于这2对引物建立的检测体系均不受杏鲍菇组织液的干扰,灵敏度高,可检测出最低3.6 pg·μL^(-1)的P.pleuroti基因组DNA;应用这2种体系对接种P.pleuroti 48 h后的杏鲍菇样品进行了检测,最低可检测出子实体内100 cfu的杏鲍菇枯萎病菌;此外,整体上,引物K3143F/R比K37F/R的检测效率更高。Pantoea pleuroti,blight disease pathogen of Pleurotus eryngii was used as the test material,PCR primers were designed,synthesized and screened after analyzing the sequencing results of P.pleuroti genome,and the detection effect was studied,so as to establishefficient PCR detection system for P.pleuroti and provide molecular basis for scientific control of blight disease of P.eryngii.The results showed that the specificity of K3143 f/R and K37 f/R primers were strong,only when P.pleuroti genomic DNA was used as template,the PCR products showed a specific band of 660 bp and 666 bp,respectively,while the genomic DNA of 15 strains of Pantoea,three pathogens strains of common edible fungi disease and distilled water(negative control)were used as template,the PCR products showed no band.The detection systems based on these two primer pairs were of high sensitivity and were not interfered by the tissue fluid of P.eryngii,which could detect the lowest 3.6 pg·μL^(-1) P.pleuroti genomic DNA.Furthermore,at least 100 cfu P.pleuroti could be detected when P.pleuroti had been inoculated onto the fruiting bodies of P.eryngii for 48 hours.In addition,primer K3143 F/R was of higher efficiency than that of K37 F/R.

关 键 词：杏鲍菇 枯萎病 Pantoea pleuroti 聚合酶链式反应 检测 

分 类 号：S436.421.13[农业科学—农业昆虫与害虫防治]