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本文建立了美澳型核果褐腐病菌(Monilinia fructicola)环介导等温扩增技术,并利用此技术开展了M.fructicola的检测和鉴定。采用M.fructicola SCAR分子标记的MO368特异片段为靶标序列,设计并筛选出M.fructicola的特异性LAMP引物,建立了该病菌快速诊断方法。试验结果表明,仅M.fructicola的菌株DNA扩增后呈天蓝色的阳性反应,而在其他真菌的供试菌株中均呈紫色的阴性反应。检测灵敏度可达到100 pg/μL。利用该方法对无锡机场截获的7份疑似李子褐腐病样本进行检测,结果有3份样品呈LAMP阳性反应。本文建立的M.fructicola LAMP检测方法具有灵敏度高、特异性好,简便易行等优点,为美澳型核果褐腐病菌的快速检测提供了一种新技术。
In this paper, the ring-mediated isothermal amplification of Monilinia fructicola was established in the United States and Australia. The detection and identification of M.fructicola were carried out by using this technique. Using M.fructicola SCAR-labeled MO368 specific fragment as the target sequence, a specific LAMP primer of M.fructicola was designed and screened, and a rapid diagnostic method for this bacterium was established. The results showed that M.fructicola only had a blue sky-positive reaction after amplified DNA, but a purple negative reaction in all other fungi. Detection sensitivity of up to 100 pg / μL. Using this method, seven samples of suspected plum brown rot intercepted from Wuxi Airport were tested. As a result, three samples were positive for LAMP. The M.fructicola LAMP detection method established in this paper has the advantages of high sensitivity, good specificity, simple and easy-to-use, which provides a new technology for the rapid detection of brown rust in the United States and Australia.