| 赵玉强1*, 卓可儿2, 郭云2, 朱灿灿1, 王行军3, 陆小美4, 田艳丽2, 胡白石2.薄壳山核桃疮痂病菌TaqMan荧光定量PCR检测方法的建立及应用[J].植物保护,2023,49(3):193-198. |
| 薄壳山核桃疮痂病菌TaqMan荧光定量PCR检测方法的建立及应用 |
| Development and application of TaqMan fluorescence quantitative PCR for detection of Venturia effusa |
| 投稿时间：2022-03-29 修订日期：2022-07-08 |
| DOI：DOI: 10.16688/j. zwbh. 2022163 |
| 中文关键词: 薄壳山核桃疮痂病菌 荧光定量PCR TaqMan探针 快速检测 |
| 英文关键词:Venturia effusa fluorescence quantitative PCR TaqMan probe rapid detection |
| 基金项目:常州市科技计划（农业科技支撑）（CE20222008） |
| 作者 | 单位 | E-mail | | 赵玉强1*, 卓可儿2, 郭云2, 朱灿灿1, 王行军3, 陆小美4, 田艳丽2, 胡白石2 | 1. 江苏省中国科学院植物研究所(南京中山植物园), 南京 210014
2. 南京农业大学植物保护学院, 农作物生物灾害综合治理教育部重点实验室, 南京 210095
3. 安徽凤阳县植保站, 滁州233100
4. 常州果美农业科技有限公司, 常州 213245 | zhaoyuqiang123@126.com |
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| 中文摘要: |
| 疮痂病是薄壳山核桃上最具毁灭性的病害，带菌植物材料是传播疮痂病的重要来源。准确、灵敏、快速的检测方法可为该病害流行规律调查和防控提供有力的依据。本文通过比较薄壳山核桃疮痂病菌Venturia effusa及其近似种之间的ITS序列差异, 设计了特异性引物和TaqMan探针, 建立了薄壳山核桃疮痂病菌的荧光定量PCR检测方法。特异性检测结果表明, 该方法可以检测不同地区的薄壳山核桃疮痂病菌菌株, 而对其近似种以及薄壳山核桃上的其他真菌均没有信号。本研究建立的检测方法对薄壳山核桃疮痂病菌DNA的最低检测限可达0.5 pg/μL。该方法用于田间样品检测时, 检测时间仅需1 h, 远快于常规的分离培养法。本研究建立的基于TaqMan探针的荧光定量PCR检测方法为薄壳山核桃疮痂病菌的快速检测和监测提供了有力工具。 |
| 英文摘要: |
| Venturia effusa causes pecan scab, which is the most destructive disease in pecan orchards. The fungus carrying plant materials are the important sources for transmission of the scab disease. Accurate, sensitive and rapid detection of the pathogenic fungi provides powerful measures for investigation of epidemical regularity and control of this disease. In this study, a pair of primers and a TaqMan probe based on the internal transcribed spacers (ITSs) of V. effusa and its related species were designed and synthesized for fluorescence quantitative PCR assay. The results showed that the fluorescence quantitative PCR assay facilitated detection of all V. effusa strains regardless of their geographical origins, while no signals were detected for the other fungi tested, including the closely related Venturia species and other fungi isolated from pecans. The detection limit of this assay was 0.5 pg/μL of total DNA. Compared to conventional culture method, the fluorescence quantitative PCR assay was rapid and reliable for detection of V. effusa in the field; the whole detection process can be finished in one hour, which provides a valuable tool for rapid detection and identification of V. effuse. |
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