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土壤中芸薹根肿菌qPCR检测与风险预警体系的建立与应用
Application of quantitative PCR based detection system for the risk early warning of Plasmodiophora brassicae in soil

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柴阿丽 1   李晓菁 1   张思雨 1   李兴盛 2   袁晓伟 2   石延霞 1   谢学文 1   李磊 1   李宝聚 1 *  
文摘 建立了土壤中芸薹根肿菌荧光定量PCR(qPCR)快速检测及风险预警体系。确定了芸薹根肿菌qPCR检测的特异性引物PbF/ PbR,对根肿菌质粒DNA的检测灵敏度为1.612×10~(-6) ng·μL~(-1),比普通PCR高出1 000倍;对土壤和基质中芸薹根肿菌孢子的最低检测下限均为10个·g~(-1),而土壤和基质带菌的发病阈值分别为100和1 000个·g~(-1),高于该浓度时根肿病发生风险大。本研究建立的芸薹根肿菌qPCR技术体系检测下限远低于发病阈值,可以快速、准确、定量地检测出采自四川绵阳、湖北恩施、江苏无锡、山东青岛、辽宁沈阳、山西运城、内蒙古巴彦淖尔和宁夏固原等8个地区的27份田间土壤中芸薹根肿菌的数量,实现对十字花科根肿病的监测预警,为制定产前病害防控方案提供依据。
其他语种文摘 In this study, a rapid fluorescence quantitative PCR(qPCR)detection and the risk early warning system for Plasmodiophora brassicae in soil was established. The minimum detection threshold for P. brassicae DNA with the specific primer PbF/ PbR was 1.612×10~(-6) ng·μL~(-1), which was 1 000 times higher than the sensitivity in that of conventional PCR. The minimum detection limit of qPCR for P. brassicae was 10 spores·g~(-1) in artificially infested substrate and soil, which was far lower than the threshold for causing clubroot disease, with 100 spores·g~(-1) in substrate and 1 000 spores·g~(-1) in soil, respectively. The qPCR assay was used to quickly and accurately quantify P. brassicae in 27 field soil samples in Sichuan, Hubei, Jiangsu, Shandong, Liaoning, Shanxi Provinces, and Inner Mongolia and Ningxia Autonomous Regions, which provided a basis for formulating disease prevention and control strategies.
来源 植物病理学报 ,2022,52(6):967-975 【核心库】
DOI 10.13926/j.cnki.apps.000774
关键词 芸薹根肿菌 ; 实时荧光定量PCR ; 检测 ; 风险预警
地址

1. 中国农业科学院蔬菜花卉研究所, 北京, 100081  

2. 山东省华盛农业集团股份有限公司, 青州, 262500

语种 中文
文献类型 研究性论文
ISSN 0412-0914
学科 植物保护
基金 "十四五"国家重点研发计划项目 ;  中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS) ;  大宗蔬菜产业技术体系项目
文献收藏号 CSCD:7415491

参考文献 共 27 共2页

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引证文献 1

1 周游 实时荧光定量PCR技术在湖南油菜根肿病菌检测中的应用 植物保护,2023,49(4):233-238,275
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