Department of Microbiology, Montana State University, Bozeman, Montana 59717.
Appl Environ Microbiol. 1991 Feb;57(2):553-6. doi: 10.1128/aem.57.2.553-556.1991.
Gaeumannomyces graminis, the causative agent of take-all disease of wheat, barley, and oats, was detected in infected wheat seedlings by using the polymerase chain reaction to amplify Gaeumannomyces-specific DNA fragments. Nested primers and two rounds of amplification were used to amplify two fragments, approximately 287 and 188 bp in size, from G. graminis-infected wheat seedlings. The use of nested primers greatly decreased the number of nonspecific amplification products. Polymerase chain reaction products were not obtained with DNA from seedlings infected with several other phytopathogenic fungi or with DNA from uninfected seedlings. Amplified products were visualized on agarose gels, and their identities were confirmed by DNA hybridization. This method did not require culturing the fungus and has potential for detecting G. graminis in infested wheat, barley, or oat fields.
禾顶囊壳,引起小麦、大麦和燕麦全蚀病的病原菌,通过聚合酶链反应(PCR)扩增禾顶囊壳特有的 DNA 片段,在受感染的麦苗中被检测到。巢式引物和两轮扩增,从感染禾顶囊壳的麦苗中扩增出大约 287 和 188bp 大小的两个片段。使用巢式引物大大减少了非特异性扩增产物的数量。用来自受几种其它植物病原真菌感染的麦苗或未感染麦苗的 DNA 进行 PCR 时,未获得扩增产物。扩增产物在琼脂糖凝胶上可见,并通过 DNA 杂交确认其身份。这种方法不需要培养真菌,并且有可能在受侵染的小麦、大麦或燕麦田检测到禾顶囊壳。