摘要
为确保对检疫性洋葱粉色根腐病病原土栖棘壳孢Setophoma terrestris进行准确快速的检测鉴定,本研究依据土栖棘壳孢模式菌株与近缘种在ITS序列上的差异位点,建立了该病菌基于TaqMan MGB探针的实时荧光定量PCR检测方法。该方法整个反应过程仅需约1h,其最低检测限量可达1 pg/μL;优化后的反应条件下,引物终浓度为0.6μmol/L,探针终浓度为0.6μmol/L。该方法具有高度的准确性、快速性和灵敏度,不仅适用于纯培养菌株的检测,也能检测洋葱组织样品中的土栖棘壳孢,为农林业生产和海关检验检疫提供了技术手段。
To accurately and rapidly detect the quarantine fungus Setophoma terrestris,a species-specific real-time polymerase chain reaction assay was developed in this study.A TaqMan-MGB probe was designed and synthesized according to the difference of rDNA ITS sequence between S.terrestris type strain and its closely related species.The whole reaction process took only about 1 h.The minimum detection limit was up to 1 pg/μL,and the optimized reaction conditions resulted in a final primer concentration of 0.6μmol/L and a final probe concentration of 0.6μmol/L.The method was highly accurate,rapid and sensitive,not only suitable for the detection of pure culture,but also for the detection of S.terrestris in onion tissue samples,providing a new reliable technological method for agroforestry production and customs inspection and quarantine.
作者
李雪莲
刘芳
段维军
LI Xuelian;LIU Fang;DUAN Weijun(Ningbo Academy of Inspection and Quarantine,Ningbo 315012,Zhejiang,China;Technical Center of Ningbo Customs District P.R.China,Ningbo 315012,Zhejiang,China;State Key Laboratory of Mycology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China)
出处
《菌物学报》
CAS
CSCD
北大核心
2024年第9期114-123,共10页
Mycosystema
基金
国家重点研发计划(2022YFF0608804)
海关总署科研项目(2022HK011)
西藏自治区重点研发计划(CGZH2023000222)
宁波市社会公益科研项目(2022S010)。
关键词
外来物种
生物安全
植物病害
检疫
检测
invasive species
biosecurity
plant disease
quarantine
detection
作者简介
Corresponding author:段维军,E-mail:weijunduan@tom.com,ORCID:0000-0001-7040-8149。
