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平菇细菌性褐斑病是一种严重危害平菇生产的病害,早期监测和防治是关键。采用平菇细菌性褐斑病病原菌托拉斯假单胞杆菌(Pseudomonas tolaasii)毒素基因的特异性引物(Pt-1A)/(Pt-1D1),通过扩增条件优化,建立了该菌的实时荧光定量 PCR(Real-time fluorescence quantitative Polymerase ChainReaction)检测及富集方法,并利用该方法完成了该菌在平菇表层的动态监测。试验结果表明,实时荧光定量 PCR 对托拉斯假单胞杆菌的检测范围为 102～ 109cfu · mL-1,在经过选择性培养基富集后,检测灵敏度进一步提高了 100 倍。利用选择性培养基富集及荧光定量 PCR 检测方法,可在病害症状未显现之前检测到病原菌,为平菇细菌性褐斑病的流行监测和早期防治奠定了技术支持。
Pleurotus ostreatus is a disease that seriously damages the production of Pleurotus ostreatus. Early monitoring and prevention are the key points. The specific primers (Pt-1A) / (Pt-1D1) of the Pseudomonas tolaasii toxin gene of the bacterial pathogens of Pleurotus ostreatus were used to establish the real-time fluorescence quantitative PCR (Real-time fluorescence quantitative Polymerase Chain Reaction) detection and enrichment methods, and the use of the method to complete the mushroom in the mushroom surface of the dynamic monitoring. The results showed that the detection range of real-time PCR was 102-109 cfu · mL-1 for Pseudomonas aeruginosa, and the detection sensitivity was further increased by 100-fold after enrichment with selective medium. By using selective culture medium enrichment and fluorescence quantitative PCR method, the pathogen can be detected before the symptoms of the disease appear, which lays the technical support for the epidemic surveillance and early prevention and control of the mushroom brown spot of Pleurotus ostreatus.