Beyer D M, O'Donnell K, Paley K, Wach M P
Department of Plant Pathology, The Pennsylvania State University, University Park 16802.
Bacterial Foodborne Pathogens and Mycology Research Unit, NCAUR, ARS, USDA, Peoria, IL, 61604-3999.
Plant Dis. 2013 Jan;97(1):142. doi: 10.1094/PDIS-07-12-0619-PDN.
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 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.
双孢蘑菇(Agaricus bisporus (Lange) Imbach)是在环境可控条件下进行商业化种植的蘑菇,是宾夕法尼亚州最具价值的作物。2011年8月,我们首次在宾夕法尼亚州切斯特县的两个商业蘑菇农场观察到一种毛霉科霉菌定殖在双孢蘑菇一个新的棕色大褐菇菌株的原基和成熟担子果上。该菌株是一个产生白色担子果的商业菌株与双孢蘑菇棕色野生型分离株的杂交种。蘑菇每周“出菇”一次。到第三次出菇时,两个农场25%的生产表面被一种生长迅速的菌丝体定殖,该菌丝体最初为白色,随后变为黄色至金棕色,最后变为灰色。被霉菌定殖的蘑菇出现凹陷、变色和坏死。通过菌丝尖端法从每个农场成熟、坏死的担子果中获得了该霉菌的两个纯培养物。这些分离株保藏于美国农业部农业研究局菌种保藏中心(NRRL,伊利诺伊州皮奥里亚),保藏编号为NRRL 54814至54815以及54818至54819。这些培养物在马铃薯葡萄糖琼脂上产生大量的气生孢子梗,呈二叉状分枝。光学显微镜检查显示,每个分枝末端是一个球形、多孢的孢子囊,有一个明显的囊轴。NRRL 54818和54819的单个培养物在相对的附属丝之间产生大的(175至250×200至250μm)、桶形、深棕色至黑色的接合孢子囊,表明它们是同宗配合的。该霉菌的形态和培养特征与大孢共头霉(Syzygites megalocarpus)(3)的描述相符,大孢共头霉是毛霉目的一个成员,据报道可定殖于多种主要为肉质的担子菌(2),包括韩国栽培的松口蘑(Tricholoma matsutake)(1)。通过对核糖体大亚基(LSU rDNA)5'端的D1和D2结构域进行PCR扩增和测序,获得了形态学鉴定结果的分子系统发育确认。这四个分离株共享相同的LSU rDNA等位基因。以NRRL 54814的部分LSU rDNA序列作为BLAST查询序列,在NCBI核苷酸数据库中进行搜索,结果显示它与大孢共头霉NRRL 6288的参考分离株AF157216.1有99.5%的同一性(3)。为了评估大孢共头霉的培养物是否能引发该病,用3.7×10个孢子囊孢子接种大褐菇和白蘑菇的菌盖。当在21至22°C、12小时光周期的湿润培养箱中培养时,大褐菇在接种部位2至3天内出现病害症状,包括变色和凹陷。从有症状的蘑菇中重新分离出大孢共头霉,其产生的菌落与原始菌落相同。相比之下,用相同方法接种大孢共头霉的白蘑菇仅出现轻微凹陷和变色。作为阴性对照,接种水的蘑菇未观察到病害症状。尽管利用“野生”遗传多样性种质培育新的商业品种是将理想性状引入栽培蘑菇的有效方法,但它也存在将新病害引入该行业的风险。参考文献:(1)K.-H. Ka等人,《韩国真菌学杂志》27:345,1999年。(2)R. L. Kovacs和W. J. Sundberg,《伊利诺伊州科学院学报》92:181,1999年。(3)K. O'Donnell,《培养中的接合菌》。帕尔弗里植物学贡献。第2号。佐治亚大学植物系,雅典,1979年。