Hypochnicium sensu lato (Polyporales, Basidiomycota) from Japan, with descriptions of a new genus and three new species.

Species of Hypochnicium (Polyporales, Basidiomycota) collected from Japan were studied on their taxonomy by morphological and phylogenetic approaches. Phylogenetic analyses based on a nrDNA LSU and ITS dataset including the Japanese specimens and other publicly available ones show that Hypochnicium is polyphyletic. Since the clade containing the type species H. bombycinum was well-supported, we defined this clade as Hypochnicium s. str., and emended Hypochnicium to include restricted taxa with only smooth basidiospores. The new genus Neohypochnicium is proposed to accommodate the remaining taxa excluded from the genus Hypochnicium s. str., which includes both species with smooth basidiospores and ornamented ones. Three new species, Gyrophanopsis japonica, N. asiaticum and N. perlongicystidiosum are described and illustrated based on morphological and phylogenetic analyses using an ITS region dataset. In addition, the following 15 new combinations are proposed: N. albostramineum, N. aotearoae, N. capitulateum, N. cremicolor, N. cystidiatum, N. geogenium, N. guineense, N. huinayense, N. michelii, N. microsporum, N. patagonicum, N. pini, N. punctulatum, N. subrigescens and N. wakefieldiae. An identification key to Japanese species of Bulbillomyces, Gyrophanopsis, Hypochnicium and Neohypochnicium is provided.

Taxonomy and ecology of Physisporinus forming synnema-like structures in freshwater environments.

Since 2011, we have collected fungi that form synnema-like structures (SSs) bearing many acanthophyses at the apex on water-splashed wood in streams in various regions of Japan. A provisional phylogenetic analysis of strains isolated from SSs based on their nrDNA sequences implied affinity with Physisporinus (Polyporales, Basidiomycota). However, it has not been reported that this genus forms SSs in freshwater habitats. We found a fungus forming not only SSs on the water-boundary part of wood but also resupinate basidiocarps with poroid hymenophores on nonsubmerged parts, and the morphological characteristics of the basidiocarps matched those of Physisporinus. Therefore, we investigated the relationship between SS-forming fungi and their sexual states by taxonomic approaches. Phylogenetic analyses based on nrDNA internal transcribed spacer (ITS) and large subunit (LSU) sequences indicated that SS-forming fungi diverged into five clades in Physisporinus. Each clade was discriminated by the color of SSs, morphology of acanthophyses, and cultural characteristics. Of the five clades, Clade 1, which consisted only of sequences of strains isolated from SSs and basidiocarps produced on rhizomorphs, was closely related to P. eminens and P. undatus, but the morphology of basidiocarps and the manner of basidiocarp development differed. Clade 5 was closely related to P. castanopsidis, P. crocatus, P. pouzarii, P. sanguinolentus, P. subcrocatus, P. tibeticus, and P. vitreus, but the basidiocarp morphology differed. Therefore, Clades 1 and 5 were described as two new species. Regarding Clades 2, 3, and 4, further taxonomic studies with additional specimens are required. SS and acanthophysis formation in wet habitats in streams and in culture could be recognized as new taxonomic and ecological characters of Physisporinus.

Two new species of Sistotrema s.l. (Cantharellales) from Japan with descriptions of their ectomycorrhizae.

We describe two new species of resupinate Sistotrema sensu lato (Cantharellales) collected in Japan: S. flavorhizomorphae and S. chloroporum. Both species have urniform basidia with more than four sterigmata and monomitic hyphal system, oil-rich hyphae in subiculum, which is typical for this genus. Sistotrema chloroporum is characterized by poroid hymenophore partly yellowish-green, basidia 4-6-spored, medium-sized basidiospores (4.5-6.5 × 3.5-6 µm), and broadleaf forest habitat. Sistotrema flavorhizomorphae is characterized by hydnoid-irpicoid hymenophore, bright yellowish rhizomorphs, basidia 6-8-spored, small basidiospores (3-3.5 × 2.5-3 µm), and pine forest habitat. Phylogenetic trees inferred from the fungal nrDNA ITS and LSU and the rpb2 sequences supported that both species were distinct and grouped with other ectomycorrhizal Sistotrema and Hydnum species, but their generic boundary was unclear. Mycorrhizae underneath basidiomes of both species were identified and described via molecular techniques. Mycorrhizae of S. chloroporum have similar characteristics to those of other Sistotrema s.l. and Hydnum species, i.e., S. confluens and H. repandum, whereas S. flavorhizomorphae has a distinct morpho-anatomy, for example, a distinct pseudoparenchymatous mantle. Comprehensive characterizations of basidiomes and mycorrhizae improve the taxonomic analysis of mycorrhizal species of Sistotrema s.l.

Novel tyrosinase inhibitors from liquid culture of Neolentinus lepideus.

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.

Ogataea paradorogensis sp. nov., a novel methylotrophic ascomycetous yeast species isolated from galleries of ambrosia beetles in Japan, with a close relation to Pichia dorogensis.

Two yeast strains isolated from galleries of ambrosia beetles in Japan and maintained in NITE Biological Resource Center (NBRC) as Pichia pini were found to represent a hitherto undescribed species. This species shows close relationship to Pichia dorogensis by the sequence analysis of the D1/D2 domain of 26S rDNA but is clearly differentiated from it by a DNA-DNA reassociation experiment. It is described as Ogataea paradorogensis sp. nov. The vegetative cells and asci of this species are surrounded with distinct capsules like P. dorogensis. One to four hat-shaped ascospores, which tend to be liberated from the asci at maturation, are formed in the ascus.