摘　要：为了快速准确地鉴定出3种不同的马铃薯（Solanum tuberosum L.）病原菌，即致病疫霉（Phytophthora infestans）、立枯丝核菌（Rhizoctonia solani）和茄链格孢菌（Alternaria solani），根据GenBank中的这3种病原菌基因序列设计特异引物，从影响多重聚合酶链式反应（PCR）扩增的因素（如退火温度和引物浓度）对反应体系进行优化，建立能够同时检测3种马铃薯病原菌的多重PCR方法。结果显示，所建多重PCR方法的检测限为40.0 fg/μL，灵敏度较高，且具有良好的特异性。该检测方法实现了同时对3种马铃薯病原菌的高效、快速的检测，提高了检测效率，降低了检测成本，在马铃薯病害的早期预防、诊断及防控中具有较高的应用潜力。
关键词：马铃薯病原菌；致病疫霉；立枯丝核菌；茄链格孢菌；多重PCR检测方法
中图分类号：S41　　　　　　   文献标志码：A　　　　　　   DOI: 10.3969/j.issn.1007-7146.2020.06.007

Abstract: A multiplex PCR assay was established for rapid and accurate detection of three potato pathogens, Phytophthora infestans, Rhizoctonia solani and Alternaria solani. According to the published gene sequences of these three pathogens in GenBank, we designed 3 pairs of specific primers. The main factors influencing the multiplex PCR efficiency, such as annealing temperature and primer concentration, were optimized. The results showed that our proposed multiplex PCR assay provided high sensitivity and extraordinary specificity. The detection limit was estimated to be approximately 40.0 fg/μL. With its high efficiency, low cost and rapidity, the multiplex PCR assay has great potential application in the early prevention, diagnosis and control of potato diseases.
Key words: potato pathogen; Phytophthora infestans; Rhizoctonia solani; Alternaria solani; multiplex PCR assay