The genera Leucoagaricus and Leucocoprinus in the Dominican Republic.

We studied species of Leucoagaricus and Leucocoprinus collected in the Dominican Republic over the past 10 years using morphological and molecular methods and carefully compared our collections with previously described neotropical taxa. Twelve new species, eight in Leucoagaricus (La. bulbiger, La. caeruleovertens, La. margaritifer, La. pegleri, La. roseovertens, La. silvestris, La. stillatus, La. turgipes) and four in Leucocoprinus (Lc. antillarum, Lc. fuligineopunctatus, Lc. microlepis, Lc. scissus) are described. Additional records of previously described taxa are also discussed, including the first molecularly annotated occurrences of Lepiota guatopoensis, Lepiota mucrocystis, and La. rubroconfusus in their putative natural habitats and of Lc. cretaceus in the neotropics. Lepiota guatopoensis and Lepiota mucrocystis are transferred here to Leucoagaricus based on their phylogenetic placement and morphological characteristics. Color photographs of fresh basidiocarps and line drawings of microscopic characters are provided for all species.

A revision of Agaricus section Arvenses with nine new species from China.

Agaricus sect. Arvenses includes numerous species that are potential candidates for cultivation, and some have high nutritional and medicinal interests. Between 2012 and 2017, 147 specimens of A. sect. Arvenses were collected in China. For this study, nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS), nuc 28S rDNA (28S), and translation elongation factor 1-alpha (tef1) sequences were used to assess species boundaries of these samples from China. Combined with morphological examination, we recognize 22 species of A. sect. Arvenses from China, of which 12 are known species, one is new record for China, and nine are proposed as new.

Medicinal potential of mycelium and fruiting bodies of an arboreal mushroom Fomitopsis officinalis in therapy of lifestyle diseases.

Fomitopsis officinalis is a medicinal mushroom used in traditional European eighteenth and nineteenth century folk medicine. Fruiting bodies of F. officinalis were collected from the natural environment of Swietokrzyskie Province with the consent of the General Director for Environmental Protection in Warsaw. Mycelial cultures were obtained from fragments of F. officinalis fruiting bodies. The taxonomic position of the mushroom mycelium was confirmed using the PCR method. The presence of organic compounds was determined by HPLC-DAD analysis. Bioelements were determined by AF-AAS. The biochemical composition of the tested mushroom material was confirmed with the FTIR method. Antioxidant properties were determined using the DPPH method, and the antiproliferative activity was assessed with the use of the MTT test. The presence of indole compounds (L-tryptophan, 6-methyl-D,L-tryptophan, melatonin, 5-hydroxy-L-tryptophan), phenolic compounds (p-hydroxybenzoic acid, gallic acid, catechin, phenylalanine), and sterols (ergosterol, ergosterol peroxide) as well as trace elements was confirmed in the mycelium and fruiting bodies of F. officinalis. Importantly, a high level of 5-hydroxy-L-tryptophan in in vitro mycelium cultures (517.99 mg/100 g d.w) was recorded for the first time. The tested mushroom extracts also showed antioxidant and antiproliferative effects on the A549 lung cancer cell line, the DU145 prostate cancer cell line, and the A375 melanoma cell line.

Four phylogenetic species of ergot from Canada and their characteristics in morphology, alkaloid production, and pathogenicity.

Four ergot species (Claviceps ripicola, C. quebecensis, C. perihumidiphila, and C. occidentalis) were recognized based on analyses of DNA sequences from multiple loci, including two housekeeping genes, RNA polymerase II second largest subunit (RPB2), and translation elongation factor 1-α (TEF1-α), and a single-copy ergot alkaloid synthesis gene (easE) encoding chanoclavine I synthase oxidoreductase. Morphological features, ergot alkaloid production, and pathogenicity on five common cereal crops of each species were evaluated and presented in taxonomic descriptions. A synoptic key was also provided for identification.

Characterization of morphological changes within stromata during sexual reproduction in Aspergillus flavus.

Aspergillus flavus contaminates agricultural products worldwide with carcinogenic aflatoxins that pose a serious health risk to humans and animals. The fungus survives adverse environmental conditions through production of sclerotia. When fertilized by a compatible conidium of an opposite mating type, a sclerotium transforms into a stroma within which ascocarps, asci, and ascospores are formed. However, the transition from a sclerotium to a stroma during sexual reproduction in A. flavus is not well understood. Early events during the interaction between sexually compatible strains of A. flavus were visualized using conidia of a green fluorescent protein (GFP)-labeled MAT1-1 strain and sclerotia of an mCherry-labeled MAT1-2 strain. Both conidia and sclerotia of transformed strains germinated to produce hyphae within 24 h of incubation. Hyphal growth of these two strains produced what appeared to be a network of interlocking hyphal strands that were observed at the base of the mCherry-labeled sclerotia (i.e., region in contact with agar surface) after 72 h of incubation. At 5 wk following incubation, intracellular green-fluorescent hyphal strands were observed within the stromatal matrix of the mCherry-labeled strain. Scanning electron microscopy of stromata from a high- and low-fertility cross and unmated sclerotia was used to visualize the formation and development of sexual structures within the stromatal and sclerotial matrices, starting at the time of crossing and thereafter every 2 wk until 8 wk of incubation. Morphological differences between sclerotia and stromata became apparent at 4 wk of incubation. Internal hyphae and croziers were detected inside multiple ascocarps that developed within the stromatal matrix of the high-fertility cross but were not detected in the matrix of the low-fertility cross or the unmated sclerotia. At 6 to 8 wk of incubation, hyphal tips produced numerous asci, each containing one to eight ascospores that emerged out of an ascus following the breakdown of the ascus wall. These observations broaden our knowledge of early events during sexual reproduction and suggest that hyphae from the conidium-producing strain may be involved in the early stages of sexual reproduction in A. flavus. When combined with omics data, these findings could be useful in further exploration of the molecular and biochemical mechanisms underlying sexual reproduction in A. flavus.

Hymenophore configuration of the oak mazegill (Daedalea quercina).

The complex hymenophore configuration of the oak mazegill (Daedalea quercina, Polyporales) is rarely quantified, although quantifications are important analytical tools to assess form and growth. We quantified the hymenophore configuration of the oak mazegill by manual counting of tubes and tubular branches and ends. Complementary measurements were made with the software AngioTool. We found that the number of tubular branches and ends varied substantially between specimens, with a positive correlation with hymenophore area (5-51 cm2). We then measured complexity as tubular branches and ends per area, and complexity was not correlated with the size of the basidiocarps. Basidiocarps from two locations were compared (Hald ege, N = 11; Hvidding krat, N = 7), and the prevalence of branches and that of ends were greater in the Hvidding krat hymenophores (P < 0.001 and P = 0.029, respectively). Additionally, lacunarity, a measure of complexity ("gappiness"), gave a higher score for the Hald ege hymenophores (P = 0.002). Lacunarity analysis of multiple species of Polyporales showed that the oak mazegill hymenophore is comparatively complex. Concerning factors that affect hymenophore complexity of the oak mazegill, we observed that greater hymenophore complexity was associated with abrupt boundaries between growth zones on the pileus surface. Several years of monitoring documented that basidiocarps can remodel to gravitational changes and heal from damage. In conclusion, intra- and interspecies differences of hymenophore configuration can be quantified. In oak mazegill, hymenophore complexity is not dependent on size per se, although abrupt borders between growth zones are associated with increased complexity. Some of the variation between basidiocarps may reflect aspects of the ecology of the individual fungus.

Taxonomic study of Endogonaceae in the Japanese islands: New species of Endogone, Jimgerdemannia, and Vinositunica, gen. nov.

Species of Endogonaceae (Endogonales, Mucoromycotina) are characterized by the formation of relatively large sporocarps and zygosporangia. Numerous species in this family remain undescribed or have unclear phylogenetic positions. In Asia specifically, the species diversity of this family is almost completely unknown. However, many mycobionts of bryophytes belonging to several novel clades in Endogonaceae have recently been identified phylogenetically. Therefore, establishing a robust taxonomic system for this family is essential. We obtained numerous sporocarps of undescribed Endogonaceae-like species from the Japanese islands. Morphological observation and multilocus phylogenetic analysis of nuc 18S rDNA (18S), nuc 28S rDNA (28S), and portions of two nuclear protein-coding regions-translation elongation factor 1-alpha (tef1) and RNA polymerase II large subunit (rpb1)-from these species resulted in the description of one new species each of Endogone and Jimgerdemannia and two new species of Vinositunica, gen. nov. Because Vinositunica is characterized by purplish sporocarps and red-wine-colored chlamydospores up to 700 µm in diameter, we emended the definition of Endogonaceae.

Structural investigation of cell wall polysaccharides extracted from wild Finnish mushroom Craterellus tubaeformis (Funnel Chanterelle).

Craterellus tubaeformis (Funnel Chanterelle) is among the most abundant wild mushrooms in Finland. Three polysaccharide fractions were sequentially extracted from the fruiting bodies of C. tubaeformis, using hot water, 2% and 25% KOH solutions, respectively, and purified. The monomer composition, molecular weight, and chemical structure were determined using chromatographic and spectroscopic methods. Thermogravimetric analysis was performed as well. The hot water extract consisted mainly of high-molecular weight → 2,6)-α-Man-(1 → and → 6)-α-Gal-(1 → chains, covalently bound to proteins. The alkali extracts consisted of acidic → 6)-ß-Glc-(1→, with branches of short → 3)-ß-Glc-(1 → chains or single ß-Glc residues. The use of alkali influenced the glycosidic linkages, molecular mass and thermal stability of the polysaccharide fractions. The use of KOH 2% increased the amount of low molecular weight polysaccharides, resulting in bimodal molecular weight distributions, with little impact on the thermal stability. Conversely, extraction with KOH 25% provided low molecular weight polysaccharides with substantially reduced thermal stability.

A novel STRIPAK complex component mediates hyphal fusion and fruiting-body development in filamentous fungi.

The STRIPAK complex is involved in growth, cell fusion, development and signaling pathways, and thus malfunctions in the human STRIPAK complex often result in severe neuronal diseases and cancer. Despite the high degree of general conservation throughout the complex, several STRIPAK complex-associated small coiled-coil proteins of animals and yeasts are not conserved across species. As there are no data for filamentous ascomycetes, we addressed this through affinity purification with HA-tagged striatin ortholog PRO11 in Sordaria macrospora. Combining the method with liquid chromatography-mass spectrometry, we were able to co-purify STRIPAK complex interactor 1 (SCI1), the first STRIPAK-associated small coiled-coil protein in filamentous ascomycetes. Using yeast two-hybrid experiments, we identified SCI1 protein regions required for SCI1-PRO11 interaction, dimerization of SCI1 and interaction with other STRIPAK components. Further, both proteins PRO11 and SCI1 co-localize with the nuclear basket protein SmPOM152 at the nuclear envelope. Expression of the gene sci1 occurs during early developmental stages of S. macrospora, and the protein SCI1 in combination with PRO11 is required for cell fusion, vegetative growth and sexual development. The results of the present study will help to understand the underlying molecular mechanisms of STRIPAK signaling and function in cellular development and diseases in higher eukaryotes.

The genome sequence of the commercially cultivated mushroom Agrocybe aegerita reveals a conserved repertoire of fruiting-related genes and a versatile suite of biopolymer-degrading enzymes.

BACKGROUND: Agrocybe aegerita is an agaricomycete fungus with typical mushroom features, which is commercially cultivated for its culinary use. In nature, it is a saprotrophic or facultative pathogenic fungus causing a white-rot of hardwood in forests of warm and mild climate. The ease of cultivation and fructification on solidified media as well as its archetypal mushroom fruit body morphology render A. aegerita a well-suited model for investigating mushroom developmental biology. RESULTS: Here, the genome of the species is reported and analysed with respect to carbohydrate active genes and genes known to play a role during fruit body formation. In terms of fruit body development, our analyses revealed a conserved repertoire of fruiting-related genes, which corresponds well to the archetypal fruit body morphology of this mushroom. For some genes involved in fruit body formation, paralogisation was observed, but not all fruit body maturation-associated genes known from other agaricomycetes seem to be conserved in the genome sequence of A. aegerita. In terms of lytic enzymes, our analyses suggest a versatile arsenal of biopolymer-degrading enzymes that likely account for the flexible life style of this species. Regarding the amount of genes encoding CAZymes relevant for lignin degradation, A. aegerita shows more similarity to white-rot fungi than to litter decomposers, including 18 genes coding for unspecific peroxygenases and three dye-decolourising peroxidase genes expanding its lignocellulolytic machinery. CONCLUSIONS: The genome resource will be useful for developing strategies towards genetic manipulation of A. aegerita, which will subsequently allow functional genetics approaches to elucidate fundamentals of fruiting and vegetative growth including lignocellulolysis.

Two new species in the Suillus spraguei complex from China.

Suillus spraguei, synonym S. pictus, has been reported from eastern North America and eastern Asia associated with Pinus subgenus Strobus. Published phylogenetic analyses of rRNA internal transcribed spacer (ITS) sequence and population genetic studies indicated that S. spraguei as currently circumscribed might contain several geographically distinct species. This study examined this possibility through a multigene analysis of S. spraguei specimens from eastern North America and eastern Asia. These specimens were associated with Pinus strobus, P. koraiensis, P. armandii, and P. kwangtungensis. The multigene analysis included three genomic regions: the genes for translation elongation factor 1α (TEF1) and RNA polymerase II largest subunit (RPB1), and the nuc rRNA segments ITS1-5.8S-ITS2 (ITS) and 28S D1-D2 domains (28S). This study confirms that the S. spraguei complex consists of at least three cryptic species: S. spraguei sensu stricto associated with P. strobus in eastern North America; S. phylopictus associated with multiple species in Pinus subgenus Strobus (5-needle pines) throughout China and Japan; and S. kwangtungensis, currently found only in P. kwangtungensis forests in southeastern China. A third new species from Japan and Korea was suggested based on ITS phylogeny. Morphologically, S. spraguei and S. phylopictus resemble each other, whereas S. kwangtungensis is covered with more floccose scales. The new species add to the knowledge of macrofungal diversity in eastern Asia and highlight the necessity of comparing broadly distributed species complexes using morphological, molecular, and ecological data.

Two new species of Phylloporus from Bangladesh, with morphological and molecular evidence.

Two new species of Phylloporus from Bangladesh forests dominated by Shorea robusta are described and illustrated, based on morphological and molecular phylogenetic evidence. Phylloporus attenuatus is characterized by its wine-red to brownish-red pilei, decurrent lamellae that stain blue, downwards tapering stipe, a trichodermium pileipellis, and ellipsoidal-ovoid to ellipsoidal basidiospores (7-8 × 4-5 µm). Phylloporus catenulatus is characterized by its small-sized basidiomata with reddish-brown pilei, narrowly decurrent with distant lamellae that stain blue, a subepithelium pileipellis, and ellipsoidal to ellipsoidal-ovoid basidiospores (8-9.5 × 4-5 µm). Both new species of Phylloporus are distinctive from any known taxa based on phylogenetic analyses of partial sequences of the nuc rDNA internal transcribed spacer (ITS; including ITS1-ITS2 but not 5.8S) region, partial sequence of nuclear ribosomal large subunit (28S) domains D1 and D2, and translation elongation factor 1-α (TEF1α). Both species are compared with phenotypically similar taxa and illustrated with line drawings and photographs. A key to the species of Phylloporus known from Bangladesh is provided.

Phylogeny and diversity of Fomitiporella (Hymenochaetales, Basidiomycota).

Fomitiporella accommodates polypores producing annual to perennial basidiocarps with an indistinct subiculum (very thin to almost lacking), mostly a dimitic hyphal structure, lacking any kind of setae, with brownish, thick-walled basidiospores, and causing a white rot. Previously, only a few samples of Fomitiporella were studied on the basis of morphological and nuc 28S rDNA (28S)-based phylogenetic analyses. In this study, we made a comprehensive study on Fomitiporella on the basis of collections from Central America, USA, Europe, and China. The phylogenetic analysis, including 28 nuc 28S rDNA and 29 nuc rDNA ITS1-5.8S-ITS2 (internal transcribed spacer [ITS]) sequences newly generated, discovered 14 new lineages. Combined with morphological evidence, 4 new lineages are described and illustrated as new species, viz., Fomitiporella americana, F. micropora, F. sinica, and F. subinermis; 10 other new lineages, each with a single collection, are still treated as unidentified taxa; three new combinations, viz., Fomitiporella tenuissima, F. chinensis, and F. resupinata, are proposed. In addition, F. inermis is redescribed. A key to the 12 known species of Fomitiporella is provided.

Unisexual reproduction of Cryptococcus gattii.

Cryptococcus gattii is a basidiomycetous human fungal pathogen that typically causes infection in tropical and subtropical regions and is responsible for an ongoing outbreak in immunocompetent individuals on Vancouver Island and in the Pacific Northwest of the US. Pathogenesis of this species may be linked to its sexual cycle that generates infectious propagules called basidiospores. A marked predominance of only one mating type (α) in clinical and environmental isolates suggests that a-α opposite-sex reproduction may be infrequent or geographically restricted, raising the possibility of an alternative unisexual cycle involving cells of only α mating type, as discovered previously in the related pathogenic species Cryptococcus neoformans. Here we report observation of hallmark features of unisexual reproduction in a clinical isolate of C. gattii (isolate 97/433) and describe genetic and environmental factors conducive to this sexual cycle. Our results are consistent with population genetic evidence of recombination in the largely unisexual populations of C. gattii and provide a useful genetic model for understanding how novel modes of sexual reproduction may contribute to evolution and virulence in this species.

Mycelial growth rate and macro- and micromorphological characteristics of medicinal species of genus Ganoderma (higher Basidiomycetes) from Iran.

Mycelial growth rate is a distinguishing quality that demonstrates continuous variation in different isolates collected from various hosts and locations. The objectives of this research were (1) to reinvestigate the previous identification of Iranian species, and (2) to recognize the best native isolate(s) for cultivation of different Ganoderma species. Of 78 samples collected from different hosts and sites, only 43 mycelia could be purified and examined for further study. Growth rate (GR; Δd/Δt) and growth coefficient (GC; dgh/t) were analyzed by growing isolate culture on 2% malt-extract agar medium (pH 5.5) incubated at 25°C. Macro- and micromorphological studies on mycelia and fruiting bodies such as basidiospore and cutis microcharacters as well as fruiting body quality were used for precise identification. Results revealed that samples belonged to 4 species: G. lucidum, G. applanatum, G. resinaceum, and G. australe. Among all samples, the isolate morphologically identified as G. applanatum showed the best GR (12 mm/day) and good GC (128 mm/day), followed by the 2 other isolates identified as G. resinaceum (GRs and GCs of 11 and 55 mm/day and 10.9 and 43.6 mm/day, respectively).

Biological characteristics of teleomorph and optimized in vitro fruiting conditions of the Hoelen medicinal mushroom, Wolfiporia extensa (Higher Basidiomycetes).

Wolfiporia extensa is a basidiomycetous brown rot fungus and is of well-known medicinal import in China, Japan, and other Asiatic countries. Fruiting body induction is of major relevance for basic biological research and for their use in industrial applications. Based on the evaluation of the effects of temperature, time in the dark before induction and culture, and wounding treatment on fruiting, this report describes the most efficient protocol for inducing fruiting of W. extensa growing on agar plates. Furthermore, several biological characteristics of teleomorph, such as the locations of hymenium, the configuration of basidiospores and primary mycelia, and events involved in basidiosporogenesis in W. extensa, were analyzed for the first time using fluorescence microscopy. The results showed that the hymenium born on both sides of the wall of the honeycomb-like structure on the surface of fruiting bodies and the hymenophoral trama situated in the middle. Each basidia has 4 binuclear basidiospores, and the primary mycelia are multinucleate without clamp connections. These results broaden our knowledge about this brown rot fungus and promote further studies of the sexual reproduction, fruiting body development, and advancement of breeding program, new topics related to the contents of pharmacologically active substances in W. extensa fruiting bodies.

Evolution of habitat preference and nutrition mode in a cosmopolitan fungal genus with evidence of interkingdom host jumps and major shifts in ecology.

Host jumps by microbial symbionts are often associated with bursts of species diversification driven by the exploitation of new adaptive zones. The objective of this study was to infer the evolution of habitat preference (decaying plants, soil, living fungi, and living plants), and nutrition mode (saprotrophy and mycoparasitism) in the fungal genus Trichoderma to elucidate possible interkingdom host jumps and shifts in ecology. Host and ecological role shifts were inferred by phylogenetic analyses and ancestral character reconstructions. The results support several interkingdom host jumps and also show that the preference for a particular habitat was gained or lost multiple times. Diversification analysis revealed that mycoparasitism is associated with accelerated speciation rates, which then suggests that this trait may be linked to the high number of species in Trichoderma. In this study it was also possible to infer the cryptic roles that endophytes or soil inhabitants play in their hosts by evaluating their closest relatives and determining their most recent ancestors. Findings from this study may have implications for understanding certain evolutionary processes such as species radiations in some hyperdiverse groups of fungi, and for more applied fields such as the discovery and development of novel biological control strategies.

Genes encoding FAD-binding proteins in Volvariella volvacea exhibit differential expression in homokaryons and heterokaryons.

Flavin adenine dinucleotide (FAD)-binding proteins play a vital role in energy transfer and utilization during fungal growth and mycelia aggregation. We sequenced the genome of Volvariella volvacea, an economically important edible fungus, and discovered 41 genes encoding FAD-binding proteins. Gene expression profiles revealed that the expression levels of four distinctly differentially expressed genes in heterokaryotic strain H1521 were higher than in homokaryotic strains PYd15 and PYd21 combined. These observations were validated by quantitative real-time PCR. The results suggest that the differential expression of FAD-binding proteins may be important in revealing the distinction between homokaryons and heterokaryons on the basis of FAD-binding protein functionality.

A common new species of Inocybe in the Pacific Northwest with a diagnostic PDAB reaction.

A species of Inocybe common in Washington, Oregon and British Columbia is documented and described as new. The species, I. chondroderma, is characterized by these features: pileus with a fulvous disk and ochraceous to chamois margin, presence of a cortina, densely mycelioid stipe base, smooth spores and fall phenology. The most reliable and distinctive feature of the species is a blue-green or turquoise reaction in response to application of a solution of p-dimethylaminobenzaldehyde (PDAB), indicating the presence of what is most likely an indole alkaloid. PDAB use provides a quick and diagnostic character easily implemented in a laboratory setting. ITS sequences from recent collections of I. chondroderma and from materials collected in the 1940s in Washington and Oregon fully match numerous mislabeled sequences from specimens in British Columbia and Oregon. The species is most closely related to an unclarified taxon from Colorado and Japan (I. cf. chondroderma) and a rare European species, I. subnudipes. Nine different species names in Inocybe and one in Hebeloma attributed to I. chondroderma based on GenBank BLASTN searches of the ITS locus match with 99-100% similarity, reinforcing concerns about taxonomic inaccuracies in public DNA sequence databases. A complete morphological description, illustrations and phylogenetic assessment are provided.

Taxonomy and phylogeny of the genus Megasporoporia and its related genera.

Taxonomic and phylogenetic studies on Megasporoporia s.l. were carried out. Phylogenetic analysis based on ITS and nLSU sequences showed that Megasporoporia s.l. belonging to the core polyporoid clade, however, it is not monophyletic, and four clades were recognized. The Megasporoporia s.s. clade includes M. setulosa and two new species, M. bannaensis and M. minor spp. nov. Two monophyletic clades were segregated from Megasporoporia s.l., and two new genera were established. Megasporia gen. nov. is composed of M. cystidiolophora, M. ellipsoidea, M. hexagonoides, M. major, M. violacea, and two new species, M. guangdongensis and M. hengduanensis spp. nov. Megasporoporiella gen. nov. including M. cavernulosa, M. rhododendri, M. subcavernulosa, and two new species, M. lacerata and M. pseudocavernulosa spp. nov. Megasporoporia quercina grouped with Grammothele fuligo in the Grammothele clade, so it is transferred to Grammothele and a new combination, G. quercina, is proposed. The main morphological characters of Megasporoporia and the two new genera are discussed, and identification keys to the three genera are provided.