Annulohypoxylon stygium is an ascomycete that helps Tremella fuciformis produce the fruiting body in wild state or artificial cultivation. Although the interaction between these two fungi is well known, the underlying molecular mechanism is limited. In this study, the 981 bp and 886 bp promoter sequences of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene and α-tubulin gene have been cloned, respectively. Sequence analysis showed that gpd promoter contained nine CAAT boxes, and single TGACG-motif, TATA box, ABRE motif, STRE motif, MYB motif, and W box. The α-tubulin promoter included eight CAAT boxes, three STRE, two TATA boxes and MYB, single Box 4, CAT-box, CCAAT-box, TGA-element, and ABRE. Subsequently, we evaluated the promoters' function by constructing four vectors pGEH, pGRH, pTEH, and pTRH to drive fused enhanced green fluorescent protein and hygromycin B phosphotransferase (egfp-hph) or red fluorescent protein and hygromycin B phosphotransferase (rfp-hph) expression under the control of gpd or α-tubulin promoters in A. stygium. The integration of the transformed DNA into A. stygium genome was verified by PCR, Southern blot, fluorescence microscopy, and quantitative real-time PCR (qRT-PCR). All the results indicated that the two promoters could drive egfp-hph and rfp-hph expression. This result could provide help in gene functional studies by using gpd and α-tubulin promoters to direct gene over-expression or build dual promoter silencing systems.

The effects of bacteria isolated from ectomycorrhizas on sporocarp production of Laccaria parva were examined in vitro. Three bacterial strains of Bradyrhizobium were selected on the basis of their affinity for L. parva LL02 in previous confrontation tests: a positive or a negative strain that significantly increased or decreased hyphal extension areas, and a neutral strain that did not have significant effects on them. Mycelia of LL02 and a suspension of each bacterial strain were inoculated onto a surface-sterilized pine seedling in a glass bottle and then incubated for 3mo in an illuminated incubator. Plant biomass did not differ significantly among the treatments, but the level of ectomycorrhizal colonization was low in the treatment inoculated with a negative strain (Neg treatment). The frequency with which mature sporocarps occurred was higher in treatments inoculated with positive (Pos treatment) or neutral strains (Neu treatment) compared with control and Neg treatments. The ratio of biomass accounted for by mature sporocarps was low in control and Neg treatments, and high in Pos and Neu treatments. These results indicate that bacteria colonizing ectomycorrhizas affect the production of sporocarps and that the affinity between fungi and bacteria is likely to underlie this interaction.

Bioactive compounds of endophytic fungus Trichoderma polyalthiae were extracted from culture broth media. The crude extracts showed strong antimicrobial activity against human pathogens. Biologically active compounds were isolated and purified by chromatographic methods. The structures of the pure compounds were elucidated by spectroscopic methods. They were identified as Violaceol I and Violaceol II. These compounds were detected as secondary metabolites produced by this genus for the first time. Violaceol I and II had a broad spectrum of antimicrobial activity against human pathogens, including Gram-positive bacteria (Staphylococcus saprophyticus, Staphylococcus aureus, Methicillin-Resistant S. aureus, Bacillus subtilis, Bacillus cereus) and Gram-negative bacteria (Salmonella typhimurium, Shigella sonnei) and Candida albicans. Violaceol I exhibited Minimal Inhibitory Concentration (MIC) values (<9.765−156.25 µg/mL) that were higher than Violaceol II (<9.765−312.5 µg/mL). Additionally, the MIC value of the phenol violaceol from this taxon was lower than the previous reports.

Myxomycetes inhabit coarse woody debris in varying stages of decay; however, their ecology in the dead wood of evergreen broadleaf trees is not well known. In this study, we examined the relationships between myxomycete species and the decay stage of wood from fallen trees in an evergreen broadleaf forest in Japan. Myxomycete species richness and abundance were calculated for eight stages of decay in fallen logs, according to the appearance and wood hardness of log portions. A total of 70 myxomycete species (including varieties) were found on the logs. Moderately decayed wood was the preferred habitat of myxomycetes (57 species; 81% of the total) and most species inhabited moist decayed wood. Analysis by nonmetric multidimensional scaling enabled the differentiation of myxomycete assemblages, with five groupings recognized across the progression of decay. Forty-two species preferred a particular decay stage, represented by the decay index. Physarum viride and Stemonitis splendens particularly preferred the less-decayed wood and Stemonitopsis typhina var. similis especially inhabited the well-decayed wood. Species from the order Physarales dominated the less-decayed wood, whereas Trichiales and Liceales species dominated the softer well-decayed wood. Myxomycetes diversity was high in and varied among logs with various stages of decay in a typical Japanese evergreen forest.

Recently published phylogenetic analyses of the Golovinomyces orontii complex revealed that three different species of the genus Golovinomyces may occur on cucurbitaceous hosts, viz., G. bolayi, G. orontii s. str., and G. tabaci. Owing to its morphological characteristics, Erysiphe cucurbitacearum (≡ G. cucurbitacearum) was tentatively reduced to synonym with G. tabaci. However, final conclusions on the identity and status of E. cucurbitacearum, described from China on Cucumis sativus, as putative synonym of G. tabaci required a phylogenetic confirmation and epitypification with ex-epitype sequences. Therefore, a sample of G. cucurbitacearum collected on C. sativus in Xinjiang Uygur Autonomous Region, China, in 2014 (HMJAU-PM91862) has been sequenced and is designated as epitype of the latter species, since its holotype material (HMAS 40016, collected in 1954) turned out to be too old for molecular examinations. As a result of morphological examinations and phylogenetic analyses based on ex-epitype ITS and 28S rDNA sequences, the recently assumed synonymy of G. cucurbitacearum, found on C. sativus in China, with G. tabaci could be confirmed. In addition, two new host species of G. tabaci, viz., Trigonotis peduncularis (Boraginaceae) and Rubia cordifolia (Rubiaceae), were concurrently identified. Trigonotis peduncularis is the first verified boraginaceous host for G. tabaci.

Ascomata of a Metarhizium species were collected from Tochigi Prefecture, Japan. The ascomata were similar to M. kalasinense in its host preference for elaterids, olive-green clavate stromata, and obliquely immersed perithecia, but the asci and ascospores were significantly shorter in length than those of the latter species. Furthermore, phylogenetic analyses based on the internal transcribed spacer region of nuclear ribosomal DNA and the elongation factor 1-alpha gene showed that the Japanese species was phylogenetically distinctive in the M. anisopliae lineage, including M. kalasinense. Accordingly, we describe a new species, M. brachyspermum. Isolates of this species produced olive-green conidial masses on the surface of the colony. Also, hyphal anastomoses between two metulae were often observed on conidiophores.

Leptographium wushanense sp. nov., which is associated with Tomicus armandii, is described based on molecular sequence data and morphological characters. The phylogenetic analyses based on ITS2-partial LSU rDNA region, β-tubulin and elongation factor 1-α genes showed that L. wushanense formed a wellsupported clade and was closely related to L. conjunctum and L. yunnanense, and then nested within L. lundbergii complex. The species is characterized that its conidiophores were longer than those of L. yunnanense and shorter than those of L. conjunctum, while its conidia were larger than those of L. conjunctum and smaller than those of L. yunnanense. Furthermore, the colonial growth of L. wushanense is faster than those of L. conjunctum and L. yunnanense at 25 ℃ on 2% MEA. Both molecular data and morphological comparison confirmed the placement of the new species in genus of Leptographium.