Distribution of Basidiomycete Fungi Associated with Fairy Rings On Golf Putting Greens.

Traditional methods for identification of fairy ring fungi on putting greens are insufficient due to reliance on morphological characteristics of the basidiocarps, which are ephemeral and often do not reach a mature stage due to management practices. Molecular techniques may aid in detection of fairy ring species in soil. In 2007 and 2008, basidiocarps and soil samples were collected from 8 bermudagrass and 17 bentgrass greens exhibiting fairy ring symptoms in CA, FL, HI, IL, OK, NC, SC and TX. Genomic DNA was extracted from 75 unknown basidiomycete fungi. Extractions were made from mycelium isolated from puffball or mushroom context tissue, from mycelium isolated from a soil block, or through direct DNA extraction from infested soil. DNA was also extracted from 11 known isolates acquired from the ATCC, Japan, and Pennsylvania. The ITS region of rDNA was amplified and sequenced using the basidiomycete-specific primer set ITS1f/ITS4b. Phylogenetic trees were constructed with maximum parsimony with nodes evaluated by bootstrap analysis. Based on high sequence similarity with unknown samples, 20 Genbank accessions were also included in the analyses. Seven of the ITS sequences from unknown samples grouped in the mushroom clade, with three having high sequence similarity to Genbank accessions of Coprinus bisporus Lange or Coprinus cordisporus Gibbs. Sixty-eight ITS sequences from unknown samples grouped in the puffball clade. Ninety-eight percent of unknown puffball sequences have high sequence similarity to Genbank accessions and type isolates of either Vascellum curtisii Kriesel or Bovista dermoxantha (Vittad.) De Toni, two species associated with fairy ring on golf greens in Japan. Although over 60 different basidiomycete fungi have been associated with turfgrasses, relatively few have been found in this study to be specifically associated with fairy rings on golf putting greens.