苹果牛眼果腐病菌3个致病种LAMP检测方法的建立

Establishment of Loop- Mediated Isothermal Amplification Method for the Three Pathogenic Species of Neofabraea Which Caused the Bull's Eye Rot on Apple

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℃。

Objective：To establish a loop-mediated isothermal amplification（LAMP） method for detecting the three pathogenic species of Neofabraea which caused the bull＇s eye rot on apple.Methods：Based on the β-tublin of Neofabraea and universal fungus ITS genes,the LAMP primers ①,② and ③ were designed.The LAMP assays were carried out.Results：The LAMP primers③ which designed from universal fungus ITS genes had a specific amplification to the three pathogenic species of Neofabraea.The optimum temperature was 64℃.