大豆科学2024，Vol.43Issue(3)：317-325,9.DOI:10.11861/j.issn.1000-9841.2024.03.0317
基于Taq Man MGB探针的大豆北方茎溃疡病菌快速检测
TaqMan MGB-based Real-time Fluorescent PCR Method for The Rapid Detection of Diaporthe caulivora
摘要
Abstract
In order to accurately and rapidly detect the quarantine fungi Diaporthe caulivora,a species-specific real-time polymerase chain reaction(RT-PCR)assay was developed for the detection of D.caulivora.A pair of specific primers and a Taq Man-MGB probe were designed and synthesized according to the difference of ITS sequence between D.caulivora type and related species.A novel real-time fluorescent PCR was established to detect D.caulivora.The minimal detectable concentration of targeted DNA was 1.0 pg in 10 μL reaction mixture.Optimal primer concentration and probe concentration were 0.5 μmol·L-1 and 0.6 μmol·L-1,respectively.The method could be used for the detection and preliminary screening of the samples suspected of carrying D.caulivora.The new method is more accurate,sensitive and time-saving than the traditional method and is suitable for routine use.It also provides a valuable tool for early rapid sensitive detection and identification of D.caulivora in plants and gives plant protection offices the chance to react accordingly and to evaluate measures for plant disease management against D.caulivora.关键词
大豆北方茎溃疡病菌/实时荧光PCR/Taq Man-MGB探针/快速检测Key words
Diaporthe caulivora/real-time fluorescent PCR/Taq Man-MGB probe/rapid detection引用本文复制引用
李雪莲,于洪娟,朱小琼,段维军..基于Taq Man MGB探针的大豆北方茎溃疡病菌快速检测[J].大豆科学,2024,43(3):317-325,9.基金项目
国家重点研发计划(2022YFF0608804) （2022YFF0608804）
海关总署科研项目(2022HK011) （2022HK011）
宁波市社会公益科研项目(2022S010). （2022S010）
