Traditional methods for identification of fairy ring fungi rely on the morphology of mature basidiocarps, which are ephemeral and often do not reach maturity on golf greens due to management practices. From 2007 to 2009, basidiocarps and soil samples were collected from 15 hybrid bermudagrass and 30 bentgrass greens exhibiting fairy ring symptoms in California, Florida, Hawaii, Illinois, Oklahoma, North Caroline, South Carolina, and Wisconsin. Genomic DNA was extracted from 122 unknown samples. Extractions were made from mycelium isolated from puffball or mushroom tissue, from mycelium isolated from a soil block, or through direct DNA extraction from infested soil. DNA also was extracted from 16 reference isolates. The internal transcribed spacer (ITS) region of ribosomal DNA was amplified and sequenced using the basidiomycete-specific primer sets ITS1f/ITS4b and Basid0001/2R. Phylogenetic trees were constructed with the neighbor-joining algorithm, with nodes evaluated by bootstrap analysis. Most samples grouped into one of three clades corresponding to species within the family Lycoperdaceae: Arachnion album, Bovista dermoxantha, and Vascellum curtisii. Although over 60 different basidiomycetes have been associated with fairy rings in turfgrasses, relatively few species were found on golf putting greens in this study. Presently, DNA sequencing may be the most efficient method for attempting speciation of fairy ring fungi from infested soil.
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