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Molecular Plant Pathology

Use of Polymerase Chain Reaction to Detect Gaeumannomyces graminis DNA in Plants Grown in Artificially and Naturally Infested Soil. Joan M. Henson,Department of Microbiology, Montana State University, Bozeman 59717; Tresa Goins(2), William Grey(3), Donald E. Mathre(4), and Monica L. Elliott(5). (2)Department of Microbiology, Montana State University, Bozeman 59717; (3)(4)Department of Plant Pathology, Montana State University, Bozeman 59717; (5)Fort Lauderdale Research and Education Center, University of Florida, Fort Lauderdale 33314. Phytopathology 83:283-287. Accepted for publication 28 October 1992. Copyright 1993 The American Phytopathological Society. DOI: 10.1094/Phyto-83-283.

The polymerase chain reaction was used to specifically amplify DNA of Gaeumannomyces graminis, a filamentous soilborne fungus that causes root and crown disease of cereals and turfgrasses. Nested primers were used to amplify a 188-bp fragment of mitochondrial DNA from fungal and infected-plant samples, which were simply boiled to release target DNA. Fungal DNA was amplified from single boiled hyphal tips or ascospores. G. graminis DNA was also detected in boiled crowns and roots from experimentally and naturally infected wheat, rice, and St. Augustinegrass and experimentally infected bermudagrass. Inhibition of DNA amplification by soil was observed in reactions with relatively low copy numbers of target DNA sequences (i.e., up to 40 ascospores) but not in reactions with large numbers of target DNA sequences (i.e., entire perithecia).