Molecular characterization of the most common dsRNA element from Trichoderma atroviride indicated that it comprised 8,566 bp and encoded two large open reading frames (ORF1 and 2). The two ORFs were found to overlap by 46 bp with a typical (-1) slippery heptanucelotide sequence. The deduced protein sequences of ORF1 and ORF2 showed significant similarities to those of known mycoviral structural proteins and RNA-dependent RNA polymerases, respectively. Phylogenetic analysis indicated that this dsRNA is a member of a distinct species related to a group of unclassified mycoviruses; therefore, it was named Trichoderma atroviride mycovirus 1 (TaMV1).
CpBck1, an ortholog of the cell-wall integrity mitogen-activated protein kinase kinase kinase of Saccharomyces cerevisiae, was cloned and characterized from the chestnut blight fungus Cryphonectria parasitica. The CpBck1-null mutant displayed cell wall integrity-related phenotypic changes such as abnormal cell morphology and wall formation and hypersensitivity to cell wall-disrupting agents. In addition, the mutant showed severely retarded growth without any sign of normal development, such as hyphal differentiation, conidiation, or pigmentation. As the culture proceeded, the mutant colony showed sporadic sectorization. Once sectored, the sectored phenotype of robust mycelial growth without differentiation was stably inherited. Compared with the wild type, both the parental CpBck1-null mutant and the sectored progeny exhibited marked impaired virulence. The present study revealed that a mutation in a signaling pathway component related to cell-wall integrity resulted in sporadic sectorization and these sectored phenotypes were stably inherited, suggesting that this signal transduction pathway is implicated in adaptive genetic changes for sectorization.
A total of 315 fungal isolates causing green mold disease were collected from contaminated artificial logs and sawdust bags used for cultivating shiitake Lentinula edodes in Korea and were analyzed for the presence of double-stranded RNA (dsRNA). dsRNA, which was purified using dsRNA-specific chromatography and verified by dsRNA-specific RNaseIII digestion, was detected in 32 isolates. The molecular taxonomy of dsRNA-infected isolates indicated that all isolates belonged to the Trichoderma spp.. The number and size of dsRNAs varied among isolates and the band patterns could be categorized into 15 groups. Although there were seven dsRNA groups observed in multiple isolates, eight groups were found to occur in single isolates. The most common dsRNA group, group VI, which contained a band of 10 kb, occurred in 10 isolates encompassing three species of Trichoderma Partial sequence analysis of two selected dsRNA groups revealed a high degree of similarity to sequences of a RNA-dependent RNA polymerase, hypothetical protein, and polyprotein genes of other hypoviruses such as Macrophomina phaseolina hypovirus 1, Trichoderma hypovirus, and Fusarium graminearum hypovirus 2, respectively, indicating the occurrence of mycoviruses in Trichoderma spp.. Northern blot analysis suggested that many different mycoviruses, which have not been identified yet, exist in Trichoderma.
The growing popularity of traditional Korean alcoholic beverages has led to a demand for quality enhancement of the traditional starter culture nuruk, which consists primarily of wheat. Therefore, this study focused on mycoflora characterization and the temporal variations in traditional wheat-based nuruks fermented at two representative traditional temperature conditions for 30 days. Nuruk A was fermented at a constant temperature of 36°C for 30 days and nuruk B was fermented at a high initial temperature of 45°C for 10 days followed by 35°C for 20 days. The average mycoflora load in the two different nuruk conditions did not vary significantly between the 0 and 30 day cultures, and a maximum load of 8.39 log CFU/g was observed for nuruk A on culture day 3 and 7.87 log CFU/g for nuruk B on culture day 30. Within two samples, pH was negatively correlated with temporal changes in mycoflora load. The pH of nuruk A was significantly lower than that of nuruk B at all of the time points evaluated. Culture-dependent characterization led to the identification of 55 fungal isolates belonging to 9 genera and 15 species, with the most prominent genera comprising Lichtheimia, Penicillium, Trametes, Aspergillus, Rhizomucor, and Mucor. A total of 25 yeast isolates were characterized belonging to 6 genera and 7 species, the most prominent among which were Rhodotorula, Pichia, Debaryomyces, Saccharomycopsis, and Torulospora. Mycofloral community dynamics analysis revealed that both samples A and B varied considerably with respect to the fungal communities over a span of 30 days.
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