First Report of <i>Syzygites megalocarpus</i> (Mucorales) Web Mold on the Commercial Portabella Button Mushroom <i>Agaricus bisporus</i> in North America (2012)

Agaricus bisporus (Lange) Imbach mushrooms, which are cultivated commercially under environmentally controlled conditions, are the most valuable crop in Pennsylvania. In August 2011, we first observed a mucoraceous mold colonizing primordia and mature basidiocarps of a new brown portabella strain of A. bisporus at two commercial mushroom farms in Chester County, PA. This strain is a hybrid between a commercial strain producing white basidiocarps and a brown wild type isolate of A. bisporus. Mushrooms mature in weekly "flushes". By third flush, 25% of the production surface at both farms was colonized by a fast growing mycelium that was initially white, subsequently yellow to golden brown, and finally grayish. Mushrooms colonized by the mold showed pitting, discoloration, and necrosis. Two pure cultures of the mold were obtained by the hyphal tip method from mature, necrotic basidiocarps at each farm. These isolates were accessioned in the ARS Culture Collection (NRRL, Peoria, IL) as NRRL 54814 to 54815 and 54818 to 54819. The cultures produced abundant aerial sporangiophores that branched dichotomously on potato dextrose agar. Light microscopic examination revealed that each branch terminated in a globose, multispored sporangium with a conspicuous columella. Individual cultures of NRRL 54818 and 54819 produced large (175 to 250 × 200 to 250 μm), barrel-shaped, dark brown to black zygosporangia between opposed suspensors, indicating they were homothallic. Morphological and cultural characteristics of the mold matched the description of Syzygites megalocarpus (3), a member of the Mucorales reported to colonize diverse, mostly fleshy basidiomycetes (2), including cultivated matsutake (Tricholoma matsutake) in Korea (1). Molecular phylogenetic confirmation of the morphological identification was obtained by PCR amplifying and sequencing domains D1 and D2 at the 5' end of the nuclear ribosomal large subunit (LSU rDNA). The four isolates shared an identical LSU rDNA allele. A search of the NCBI nucleotide database, using a partial LSU rDNA sequence from NRRL 54814 as the BLAST query, revealed that it shared 99.5% identity with AF157216.1, a reference isolate of S. megalocarpus NRRL 6288 (3). To assess whether cultures of S. megalocarpus could induce the disease, caps of portabella and white button mushrooms were inoculated with 3.7 × 10<sup>6</sup> sporangiospores. When incubated in moist chambers at 21 to 22°C with a 12-h photoperiod, disease symptoms developed in 2 to 3 days on portabella that included discoloration and pitting at the site of inoculation. S. megalocarpus was reisolated from the symptomatic mushrooms and produced a colony identical to the original. By comparison, white button mushrooms inoculated with S. megalocarpus, using the same method, only showed minor pitting and discoloration. Disease symptoms were not observed on mushrooms inoculated with water as a negative control. Although development of new commercial varieties derived using "wild" genetically diverse stocks is an effective way to introduce desirable traits into cultivated mushrooms, it carries the risk of introducing new diseases into the industry. References: (1) K.-H. Ka et al. Korean J. Mycology 27:345, 1999. (2) R. L. Kovacs and W. J. Sundberg. Trans. Il. State Acad. Sci. 92:181, 1999. (3) K. O'Donnell. Zygomycetes in culture. Palfrey Contributions in Botany. No. 2. Department of Botany, University of Georgia, Athens, 1979.