引起黄芪根腐病的锐顶镰刀菌的RPA检测方法  

Recombinant polymerase amplification method for detection of Fusarium acuminatum causing Astragalus membranaceus root rot

作　　者：王春伟 李广信[2] 周书悦 左敏 王琳 苏健 王燕 Wang Chunwei;Li Guangxin;Zhou Shuyue;Zuo Min;Wang Lin;Su Jian;Wang Yan(College of Plant Protection,Shanxi Agricultural University,Taigu Shanxi 030801,China;College of Agriculture,Shanxi Agricultural University,Taiyuan Shanxi 030031,China;Taiyuan Customs District,Taiyuan,Shanxi 030021,China)

机构地区：[1]山西农业大学植物保护学院,山西太谷030801 [2]山西农业大学农学院,山西太原030031 [3]太原海关,山西太原030021

出　　处：《中国植保导刊》2024年第9期5-9,共5页China Plant Protection

基　　金：国家重点研发计划(2019YFC1710800);山西农业大学学术恢复科研专项项目(2020xshf05);山西农业大学科技创新基金项目(2016YJ11,2016YJ12);山西省现代农业产业技术体系建设专项资金(2021-11)。

摘　　要：黄芪根腐病是可由多种镰刀菌引起的土传病害,锐顶镰刀菌为主要病原菌。根据锐顶镰刀菌转录延伸因子(TEF-1α)的特异性区段设计引物,通过优化反应温度、反应时间、灵敏度和特异性试验,以及开展对实际样品的检测,建立了锐顶镰刀菌的的重组酶聚合酶扩增(RPA)检测方法。该方法的最优反应条件为39℃、30 min,能够特异性检测锐顶镰刀菌,检测灵敏度为10μg/mL;该方法应用于实际样品检测,具有检测速度快、灵敏度高、特异性强的优点,有利于黄芪根腐病的病原监测与病害的早期诊断。Astragalus membranaceus root rot is a serious soil-borne disease caused by Fusarium spp..Of these,F.acuminatum was found to be the major pathogen associated with this disease.Specific primers for F.acuminatum were designed according to the specific region of transcription elongation factor-1α(TEF-1α).A recombinant enzyme polymerase amplification(RPA) detection method was developed based on reaction temperature and time optimization,sensitivity and specificity testing,and the detection of diseased samples.The results showed that the optimal reaction conditions for this method were 39 ℃ for 30 min,which enabled the specific detection of F.acuminatum,and the detection sensitivity could reach 10 μg/mL.This RPA method could be used for the detection of infected samples in the field,which had the advantages of a rapid detection speed,high sensitivity and strong specificity,and was conducive to the pathogen surveillance and early diagnosis of A.membranaceus root rot.

关 键 词：黄芪根腐病 锐顶镰刀菌 重组酶聚合酶扩增 分子检测 

分 类 号：S435.672[农业科学—农业昆虫与害虫防治]