Kozue Sotome

Trichoderma mienum sp. nov., isolated from mushroom farms in Japan

During an investigation of Hypocrea/Trichoderma species inhabiting mushroom bedlogs, we found five strains of an undescribed species from a culture collection. These were analyzed using a combined approach, including morphology of holomorph, cultural studies, and phylogenetic analyses of the rRNA gene cluster of the internal transcribed spacer region, translation elongation factor 1-α, and RNA polymerase subunit II gene sequences. Distinctive morphological characters include stromata with green ascospores produced on potato dextrose agar medium, and Gliocladium-like to irregularly Verticillium-like conidiophores. In phylogenetic analyses, this species belongs to the Semiorbis clade, but its morphological characteristics do not match the other members of this clade. Based on morphological observations and phylogenetic analyses, we describe this as a new species, Trichoderma mienum, representing its Hypocrea teleomorph and Trichoderma anamorph.

Taxonomic study of Favolus and Neofavolus gen. nov. segregated from Polyporus (Basidiomycota, Polyporales)

We present a taxonomic study of ‘group Favolus’ and related species in Polyporus. Phylogenetic analyses of nurLSU and ITS regions revealed that the infrageneric ‘group Favolus’ is divided into two main clades. Fungi within the group share laterally stipitate basidiocarps, with non-crustose stipe surfaces, and are distinguishable by the morphology of the pileus surface. One clade is characterized by species with hyaline to brown cutis, composed of hyaline to brown agglutinated generative hyphae. The other clade accommodates species with radially striate pileus, and lacks any distinct cutis of agglutinated hyphae. We propose Neofavolus gen. nov., typified by N. alveolaris, for the former clade, and revise the genus Favolus, typified by F. brasiliensis, for the latter clade. Neofavolus includes N. mikawai and N. cremeoalbidus sp. nov., known only from temperate eastern Asia, in addition to N. alveolaris. Favolus includes members of the Polyporus grammocephalus complex, the P. tenuiculus complex, and P. pseudobetulinus. We reveal that the polypore known as ‘P. grammocephalus’ in Asia includes F. acervatus and F. emerici (= P. grammocephalus), whereas ‘P. tenuiculus’ includes three distinct species; F. brasiliensis from tropical America, and F. spathulatus and F. roseus from tropical Asia. Detailed descriptions and illustrations are provided for the accepted species in Favolus and Neofavolus.

Taxonomy and phylogenetic position of Fomitiporia torreyae, a causal agent of trunk rot on Sanbu-sugi, a cultivar of Japanese cedar (Cryptomeria japonica) in Japan

Trunk rot poses a substantial threat to Sanbu-sugi, one of the most economically important cultivars of Japanese cedar (Cryptomeria japonica). The etiology of this disease, including its main agents, is incompletely known. This trunk rot was attributed to Fomitiporia (Phellinus) hartigii or F. (Phellinus) punctata. Here we phylogenetically analyzed DNA sequences of four markers from a set of strains isolated from trunk-rot symptoms and recovered a single, monophyletic clade, indicating that a single taxon is involved. This clade was identified as Fomitiporia torreyae, a species described from eastern China. This analysis also proved that trunk rot and/or dieback of other conifers and broadleaf trees, including Sawara cypress (Chamaecyparis pisifera), Japanese umbrella pine (Sciadopitys verticillata) and Japanese pear (Pyrus pyrifolia var. culta), were caused by the same species. The phylogenetic approach to Fomitiporia revealed that the F. torreyae clade was closely related to F. bannaensis but clearly distinct from F. punctata, which originally was thought to be the cause of trunk rot in Sanbu-sugi. Fomitiporia torreyae is redescribed on the basis of more than 40 specimens from multiple hosts from Japan and China. Fomitiporia juniperina comb. nov. is proposed.

Re-evaluation of Hypocrea pseudogelatinosa and H. pseudostraminea isolated from shiitake mushroom (Lentinula edodes) cultivation in Korea and Japan

Shiitake (Lentinula edodes) is the most economically important cultivated mushroom, but yields are impacted by its competitor, Trichoderma spp. We previously found two unidentified Trichoderma species growing in bedlogs and sawdust shiitake media in Korea. Here, we identify and re-describe those two species based on molecular sequence data, morphology, and culture characteristics. Well-supported clades based on phylogenetic analyses of internal transcribed spacer, translation elongation factor 1-, and RNA polymerase subunit II sequences grouped one of the unidentified Trichoderma spp. with Hypocrea pseudogelatinosa and the other with Hypocrea pseudostraminea, and their morphologies matched well with the original descriptions of the two Hypocrea species. This study reports the first phylogenetic analyses of H. pseudogelatinosa and Japanese strains of H. pseudostraminea. Based on the phylogenetic results, we redescribed these two species using modern taxonomic concepts in Hypocrea/Trichoderma.

Taxonomic study of Asian species of poroid Auriculariales

A taxonomic study of poroid Asian species in the Auriculariales was conducted using phylogenetic and morphological methods. Phylogenetic analyses based on the LSU and ITS regions reveal that Elmerina was a polyphyletic genus. Moreover, E. hexagonoides collected from Malaysia and “E. hexagonoides” from Japan formed two distinct clades within a well-supported larger clade with Protomerulius caryae. These taxa comprise a different lineage from the type species of both Protomerulius and Protodaedalea, and we conclude that they are members of the genus Aporpium. The phylogenetic position of E. cladophora (type species of Elmerina) is still unclear, and we retain Aporpium and Protodaedalea as genera distinct from Elmerina. Aporpium strigosum sp. nov. is described to represent Japanese specimens misidentified as E. hexagonoides. Aporpium is characterized by resupinate to sessile basidiocarps, tough-fleshy to leathery context, poroid hymenophore, dimitic hyphal systems with unbranched skeletal hyphae, clavate to pyriform probasidia, and ellipsoid to cylindrical or allantoid basidiospores. Aporpium strigosum is similar to A. hexagonoides, but can be differentiated by the ochraceous to straw-colored basidiocarps, strigose pileus surface with long and stiff hairs, and the larger pores, basidia, and basidiospores.

Trichoderma eijii and T. pseudolacteum, two new species from Japan

Trichoderma eijii and T. pseudolacteum, are described here as new species from Japan. These species were isolated from decaying wood in the Tomakomai Experimental Forest in Hokkaido Prefecture and from bedlogs on shiitake mushroom (Lentinula edodes) farms, respectively. The species were characterized using a combined approach that included cultural studies, holomorph morphology, and phylogenetic analyses of internal transcribed spacer and protein coding gene sequences (RNA polymerase subunit II, translation elongation factor 1-α, endochitinase, and actin). The results of phylogenetic analyses of these gene sequences indicate that T. eijii belongs to the Hamatum clade and is closely related to Hypocrea pezizoides, H. flaviconidia, and H. atroviridis/T. atroviride, from which it differs mainly in part-ascospore size and anamorphic characteristics. Trichoderma pseudolacteum, which was previously recognized as H. lactea sensu Doi, is morphologically distinct from H. lactea (= Hypocrea citrina) and is strongly supported as a separate lineage based on our phylogenetic analyses.

Assessment of decay risk of airborne wood-decay fungi II: relation between isolated fungi and decay risk

The relationship between the taxa of airborne fungi and the decay risk was investigated. Airborne fungi in 1,000 l of air were trapped on Japanese cedar disks, and incubated in a damp container kept at 26ºC. After 16-week incubation, filamentous fungi grown on the disks were isolated and DNA extracted from each isolate was amplified with the primers ITS4/ITS5. The DNA sequences of the amplified products were determined and compared to the sequence data of GenBank to determine the species or genus according to a BLAST search. This search revealed that the isolate consisted of 5 major taxa, namely Bjerkandera sp., Phanerochaete sp. (A), Phanerochaete sp. (B), Polyporales sp. Polyporus arcularius, and 6 minor ones. Statistical analysis revealed that the major taxa were trapped on the disks in similar weather conditions except for Bjerkandera sp., which was trapped at a cooler temperature. The analysis also proved the disks to which Phanerochaete spp. or Polyporales sp. were attached showed higher mass loss. It is concluded that, under these experimental conditions, related species of Phanerochaete sordida play an important role in increasing the decay risk caused by airborne wood-decay fungi.

Novel tyrosinase inhibitors from liquid culture of Neolentinus lepideus

Abstract Tyrosinase is the key enzyme that controls melanin formation in the human skin. We performed a screening of 96 extracts of mushroom cultures and fruiting bodies for examining their inhibitory activity against mushroom tyrosinase. The ethyl acetate extracts of culture filtrate of Neolentinus lepideus exhibited the strongest inhibitory activity. The active compounds 1 and 2 were purified by repeated chromatographic separations from the extract. On the basis of spectroscopic analyses, 1 and 2 were identified to be 1,3-dihydroisobenzofuran-4,5,7-triol and 5-methoxy-1,3-dihydroisobenzofuran-4,7-diol, respectively. Lineweaver-Burk plot of the enzyme reaction in the presence of 1 indicated that 1 was a potent competitive inhibitor. The respective IC50 values of 1 and 2 were 173 and 263 μg/mL. Compound 1 at 15 μg/mL suppressed melanin accumulation stimulated by α-MSH in the murine melanoma B16 cells, as well as the induced accumulation of both tyrosinase transcript and protein without inhibiting cell proliferation. We identified new tyrosinase inhibitors in the extract of culture filtrate of Neolentinus lepideus.