植物检疫2024,Vol.38Issue(2) :32-37.DOI:10.19662/j.cnki.issn1005-2755.2024.02.005

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

丁钿 牟桂萍 魏霜 李献锋 林晓红 章柱 易建平
植物检疫2024,Vol.38Issue(2) :32-37.DOI:10.19662/j.cnki.issn1005-2755.2024.02.005

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

丁钿 1牟桂萍 2魏霜 3李献锋 3林晓红 1章柱 1易建平4
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作者信息

  • 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
  • 折叠

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13fg;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.

Abstract

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 de-tect the simulated infected maize seeds and the standard curve was established.It was found that R2=0.998 8,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

Key words

Pantoea stewartii subsp.stewartii/real-time PCR/detection

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基金项目

国家重点研发计划(2022YFC2601500)

出版年

2024
植物检疫
中国检验检疫科学研究院

植物检疫

CSTPCD
影响因子:0.498
ISSN:1005-2755
参考文献量16
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