PCR Detection of the Three <i>Neofabraea</i>Pathogenic Species Responsible for Apple Bull’s Eye Rot

Apple bull’s eye rot is caused by pathogenic Neofabraea species including N. malicorticis, N. perennans and N. alba. Fruits carrying this fungal quarantine disease are prohibited from entering China. The host plants for the pathogens include several Rosaceae fruits including apple and pear. Disease symptoms and pathogen morphology are often insufficient to determine the identity of the pathogen, particularly at the species level. In the current study, we analyzed the inter-species sequence variations in the β-tublin gene, and designed specific primers to allow PCR amplification of 554 bp fragments from pathogenic Neofabraea species. The PCR products were recovered and sequenced, and Blast search was conducted using the DNA sequences in the Genbank database. The results indicated precise PCR amplification of the target sequences from the host pathogen, which allowed unambiguous identification of the species.

苹果牛眼果腐病菌3个致病种LAMP检测方法的建立

目的:建立引起苹果牛眼果腐病的明孢盘菌属3个致病种的环介导等温扩增(LAMP)检测方法。方法:以苹果牛眼果腐病菌3个致病种为目标菌进行LAMP引物设计,根据明孢盘菌属β-tublin基因设计引物对1、2;根据真菌通用ITS序列设计引物对3,并进行LAMP试验,从中筛选出最佳引物组合,并进行最适宜反应温度的筛选。结果和结论:经验证得出,由ITS序列得到的引物组合3对苹果牛眼果腐病菌的3个致病种有特异性的扩增,扩增最适温度为64℃。

PCR检测引起苹果牛眼果腐病的明孢盘菌属3个致病种

苹果牛眼果腐病是由明孢盘菌属真菌(Neofabraea alba、N.perennans、N.malicorticis和Neofabraea undescribed species)引起的,被中国列为禁止入境的检疫性真菌病害。该病菌的寄主植物包括苹果、梨在内的多种蔷薇科果树。根据症状特点和病菌形态特征很难鉴定到种。本研究根据明孢盘菌属β-tublin基因的高度保守性,设计针对明孢盘菌属3个致病种(Neofabraea alba、N.perennans、N.malicorticis)的特异性引物,进行PCR扩增及电泳检测,表明该引物对3个致病种均可扩增出554bp片段,而对其他参试菌株无特异扩增。此外,对PCR产物进行回收、测序及Blast分析,可根据序列比对结果准确地鉴定到3个致病种的水平。