TaqMan MGB-based real-time PCR method for the detection of Stagonosporopsis chrysanthemi
The pathogen Stagonosporopsis chrysanthemi has been included in the list of imported plant quarantine pests in China.At present,with the adjustment of the classification system of fungi and the implementation of the principle of"one fungus,one name",the classification status of S.chrysanthemi and related species have been greatly adjusted.In this study,a pair of specific primers SLl/SL2 and Probe-M according to the difference of ITS sequence between model strain of S.chrysanthemi and its related species were designed and synthesized for real-time fluorescent PCR assay.The specific test results showed that only S.chrysanthemi had positive amplification,while other strains and blank controls had no increased fluorescence signal.The sensitivity test results showed that the detection limit of this method was 26 fg/μL DNA in 20 μL reaction mixture.This method exhibits strong specificity,high sensi-tivity,and rapid detection speed.The entire detection process is completed within 1.5 hours,meeting the requirements for rapid quarantine and identification of S.chrysanthemi at ports.