A mycovirus was isolated from an edible mushroom, Lentinula edodes, that was suffering from a severe epidemic. Fractionation of the diseased cell extract by isopycnic centrifugation with 50% CsCl revealed that the diseased mushroom was infected by Lentinula edodes spherical virus (LeSV), a new spherical virus with a diameter of 55 nm. The particle of LeSV encapsidated the 12 kb RNA genome by a 120 kDa coat protein. BLAST analysis of the partially sequenced LeSV genome showed 95% sequence identity with a putative RNA-dependent RNA polymerase (RdRp) gene of the mycovirus HKB, which was previously reported as being a double-stranded RNA (dsRNA) element. In contrast to HKB, the RNA genome in LeSV is encapsidated by the 120 kDa coat protein. To confirm that the LeSV coat protein is encoded by the viral genome, the N-terminal amino acid sequence of the coat protein was determined. The resulting N-terminal amino acid sequence, N-SALDVAPVVPELYFXXLEV-C, was found to be located in the middle of the HKB ORF1, suggesting that the LeSV coat protein was indeed encoded by the virus. To detect LeSV in L. edodes, a primer set targeting the RdRp gene was designed based on the partial sequence of the LeSV genome. RT-PCR analysis showed that 56 of the 84 commercially available dikaryotic cultivars carry LeSV. The transmission pattern of the virus was determined by analysing basidiospores from LeSV-infected and LeSV-free fruiting bodies. Nine out of 10 basidiospores from the LeSV-infected cultivars contained the virus while the spores from the LeSV-free parent were free of LeSV, suggesting that vertical transmission is the primary mode of LeSV propagation.
Fungal contamination is a detrimental factor affecting sawdust media-based shiitake cultivation in greenhouses. During fungal monitoring of greenhouses used for shiitake cultivation, eight fungal species were isolated and identified from indoor air and mushroom flies collected in the greenhouses. The current study reported five species as new in Korea, viz. Ascochyta hordei, Discosia artocreas, Mucor nidicola, Perenniporia medulla-panis, and Pseudozyma prolifica, and confirmed two species, Penicillium charlesii and Penicillium brevicompactum, which were previously recorded in Korea without molecular taxonomic validation. The morphological characteristics and phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer rDNA region or calmodulin gene were described for all identified species.
: Korean oak wilt disease caused by Raffaelea quercus-mongolicae is vectored by the ambrosia beetle Platypus koryoensis. To prevent the spread of the disease, the beetle infested oak tree had been cut into logs, covered with plastic vinyl, fumigated with a pesticide, and stored for three years on the site where the tree was cut. This study was carried out to get information on the fungi colonizing the fumigated oak wood. Wood disk samples collected from the fumigated oak logs at two locations in the Taejo Mountain, Cheonan city, were used for fungal isolation. A total of 99 filamentous fungal isolates were obtained from the wood disk samples. Hypocrea spp., Trichoderma spp. and Penicillium spp. were identified based on morphological characteristics and nucleotide sequence analysis of translation elongation factor 1-alpha gene and ITS rDNA region. Trichoderma was the major fungal group. R. quercus-mongolicae, and P. koryoensis were not detected from the fumigated oak wood. Our work provided evidence that after three years of storage, the fumigated oak wilt-diseased logs should be no longer harmful source of oak wilt disease transmission.
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