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全世界危害柑橘的疫霉有14种,其中冬生疫霉Phytophthora hibernalis和丁香疫霉Phytophthora syringae为我国规定禁止进境的两种检疫性柑橘真菌病害,目前尚未见有此两种病原菌的同步分子检测报道.该研究根据14种柑橘疫霉的18SrRNA,ITS,heat shock protein 90(HSP 90)等基因分别设计了疫霉属的通用引物和此两种检疫性疫霉的特异引物,通过进行反应体系优化,建立了同时检测柑橘上两种检疫性疫霉的特异三重PCR检测方法,并进行了灵敏度试验.用15个柑橘疫霉菌株与甜橙的混合DNA做模板进行模拟带菌试验,结果表明该三重PCR分子检测能实现两种检疫性疫霉菌的同步特异检测,这两种检疫性柑橘疫霉提供了特异、可靠、便捷、适于口岸应用的检测方法,可有效促进柑橘类水果的快速通关.
Phytophthora hibernalis and Phytophthora syringae are 14 kinds of citrus Phytophthora hibernalis which are harmful to citrus in the world. They are two kinds of quarantine citrus fungal diseases which are banned from entry in China. There are no synchronous molecules of these two pathogens According to the 14 kinds of genes of Phytophthora citrus, 18SrRNA, ITS, heat shock protein 90 (HSP90) and other genes, we designed the universal primers of Phytophthora and the two specific primers of quarantine Phytophthora capsici, The system was optimized and a specific triplex PCR assay was developed for the simultaneous detection of two quarantine Phytophthora infestans in citrus and the sensitivity test was carried out.The mixed fungal DNA of 15 P. citruses strains mixed with sweet orange The three-dimensional PCR molecular detection can achieve simultaneous and specific detection of two quarantine Phytophthora capsici, and the two quarantine Phytophthora citrate provide a specific, reliable and convenient detection method suitable for port application, which can effectively promote the quickness of citrus fruits Clearance.