Epipactis helleborine is an orchid species that has been introduced to North America and continues to expand its range. Numerous studies have investigated the mycorrhizal specificity of these orchids in their native range and found broad associations with Ascomycota, particularly Wilcoxina. Similar studies in North America are rare. We assessed the fungal diversity associated with populations of E. helleborine, and the surrounding soil, in mixed habitats on Vancouver Island (Canada) to make comparisons with findings from the species native range. After collecting 45 rhizomes and 36 soil samples, we amplified the nuclear ribosomal internal transcribed spacer (ITS2) region to assess alpha and beta diversity. BLAST searches were conducted, and phylogenetic reconstructions of high-matching fungal sequences from other E. helleborine studies were created. We demonstrate that E. helleborine populations continue to associate with a diverse group of Ascomycota fungi, particularly Wilcoxina spp. as they continue to expand their range northward in western North America. However, we also detected a novel association among E. helleborine rhizomes and Rhexocercosporidium, a genus containing known saprophytes and plant pathogens. Our results indicate that this orchid can pair with an extensive range of fungal partners, likely by taking advantage of varying degrees of phylogenetic niche conservatism among different fungal groups, which could have wide-reaching implications in terms of further geographic expansion and introductions to new continents.
Red leaf blotch, caused by Coniothyrium glycines, is a major leaf spot disease of soybean in Africa. Because of its unique morphological characteristics, C. glycines has been placed in numerous genera since its description in 1957, and recent phylogenetic evidence has suggested that its current placement within Coniothyrium is incorrect. To provide nomenclatural stability, we reevaluated its evolutionary history and taxonomy using a phylogenomic approach. We produced draft genomes for 13 close relatives of C. glycines and generated a consensus tree from 4643 BUSCO genes in these species and related Dothideomycetes. We show that Coniothyrium is not monophyletic and C. glycines is closely related to Aristastoma oeconomicum, a once prevalent leaf spot pathogen of cowpea in the eastern United States. Similarities in morphology and disease symptomology, along with genome distance estimations and a phylogeny based on internal transcribed spacer (ITS) and partial nuclear 28S rDNA (D1-D2) data, support the transfer of C. glycines and another species, Coniothyrium dolichi, to Aristastoma. The inclusion of this species in Aristastoma expands the morphological concept of this genus to include species that produce sclerotia and have aseptate conidia. The improved systematics and genomic resources generated with this study will advance the development of diagnostics and detection assays of this high-consequence plant pathogen.
Several species of mushroom-forming fungi have been reported to trap and consume nematodes. Nematophagy is hypothesized to have evolved as a mechanism for supplementing nitrogen, analogous to carnivorous plants. In this study, we systematically reviewed all the putative nematode-trapping saprotrophic Agaricales and documented their behavior in the presence of nematodes. Hohenbuehelia mastrucata, Pleurotus citrinopileatus, and P. djamor all trapped and consumed nematodes, as reported previously. In contrast, we found that Stropharia rugosoannulata and Coprinus comatus produce structures that have been suggested to function in nematode trapping, but these fungi neither trap nor consume the nematodes. It is possible that the structures interpreted to function in nematode trapping in S. rugosoannulata and C. comatus act as defensive mechanisms against fungivory.
Six new species of Inocybaceae from North America are described as new. Most are common and widespread throughout eastern North America. Phylogenetic analyses of nuc rDNA ITS1-5.8S-ITS2 (ITS), harnessing numerous community science records, together with nuc 28S rDNA support the phylogenetic distinctiveness of each species with strong support values. Sequences of additional genetic loci, including RNA polymerase II largest subunit (rpb1) and second largest subunit (rpb2), were also produced to supplement the descriptions and strengthen future phylogenetic studies. Of these new taxa, three are described in Inocybe: I. albofusca, a smooth-spored species in the Flocculosa clade; I. communis, a nodulose-spored species in the Xanthomelas clade; and I. lincoffii, a nodulose-spored species in I. subsect. Napipedinae. Inosperma aureiavis is a member of the Maculatum clade, whereas Pseudosperma aurivetum is a member of the Umbrinellum clade. Mallocybe diabolica is a narrow-range endemic known only from southeastern Arizona and Mexico and is allied with the Heimii clade. Complete descriptions and illustrations are presented for each species.
In 2023, conifers with Rosellinia infections were discovered in Maine, New Hampshire, and Ohio. The trees displayed twig and needle blight with dense mats of mycelium. The isolates causing the infections were provisionally identified as Rosellinia mycophila or R. herpotrichoides according to current morphology-based criteria. Both species belong to the R. thelena subgroup (subg. Corrugata) of Rosellinia as defined by Petrini (2013, Bibliotheca Mycologica 205). To date, no molecular data are available for species within this subgenus. To address this gap, we sequenced several loci and conducted a multilocus phylogenetic analysis (nuc ITS1-5.8S-ITS2 rDNA [ITS], nuc 28S rDNA D1-D2 [28S] regions, β-tubulin [TUB2], and the second-largest subunit of the DNA-directed RNA polymerase II [RPB2]). Recently, several Rosellinia species were segregated to Dematophora because they group in a separate clade and form distinct asexual morphs: Rosellinia species form Geniculosporium-like asexual states, whereas Dematophora species form Dematophora-like asexual states. Our analysis shows that R. mycophila and R. herpotrichoides form a well‑supported clade adjacent to these groups and share Nodulisporium-like anamorphs. The specimens collected in this study exhibited more than 99.3% sequence similarity. In 1961, Petrak (Sydowia 15:242-246) considered a specimen later identified as R. mycophila to be a form of R. herpotrichoides with small ascospores. In 1986, Francis (Trans Br Mycol Soc 87:397-400) disagreed and argued that the two should be maintained as separate species. Our molecular data suggest limited genetic distinction between these species; however, we do not formally synonymize the taxa. Broader sampling, especially from European and type-associated material, will be needed to evaluate whether morphological features reliably delimit R. mycophila and R. herpotrichoides.
Members of the phylum Neocallimastigomycota, often referred to as anaerobic gut fungi (AGF), primarily inhabit the gut of herbivorous animals. To date, 25 AGF genera have been described, but only five harbor more than one species. Here, we report on the isolation and characterization of novel AGF isolates from the feces of a Boer goat. The isolates formed small, mostly round, dense colonies on agar roll tubes and produced flaky, loosely clumped white biomass in liquid medium. Microscopic features included monoflagellated spherical zoospores and monocentric filamentous thallus. Sporangia were spherical or ovoid, although elongated and oddly shaped sporangia were also observed. Zoospore release was observed through ruptures in the sporangial wall. Phylogenetic analysis using the ribosomal internal transcribed spacer 1 (ITS1) and the 28S nuc rDNA D1-D2 region (28S) showed the isolates to be most closely related to Joblinomyces apicalis, the only described species in the genus Joblinomyces (0.00-1.30% divergence in ITS1, 2.50-3.82% in 28S). Transcriptomic sequencing and subsequent analysis of the RPB1 (largest subunit of RNA polymerase II) region and average amino acid identity (AAI) value comparison clearly showed a high level of AAI sequence divergence (70.11-75.40%, average 72.53 ± 1.43%) when compared with all other genera within the order Neocallimastigales. Based on morphological and phylogenetic analyses, we propose to accommodate these isolates as a novel species within the genus Joblinomyces and propose the name of Joblinomyces undulatus. The type strain for this species is JC3.3.
Fungi in the phylum Ascomycota disperse their sexual spores primarily by forcible discharge from the spore sac, the ascus, in which they develop. These fungi cause some of the most devastating diseases of agricultural crops worldwide. The mechanism of spore discharge is not well understood, despite its primary role in disease spread. Using confocal microscopy, we have elucidated the ascus structure and development as related to function for spore dispersal in Fusarium graminearum, a pathogen of small grains and corn. To determine the mechanism of spore discharge in this species, we examined ascus structure in both wild-type and spore discharge mutant strains and revealed the presence of two chambers within the ascus: a stiff-walled lower chamber that supports the developing spores and an upper chamber, extendable due to the presence of a chitin network across the surface that maintains the integrity of the sac as it is pressurized. The upper chamber includes an internal chute to confine and align the spores, while pressure builds up, with a minimum amount of fluid. Elucidation of the basis of spore dispersal for a member of this major group of fungi provides a solid beginning for exploring forcible discharge throughout this phylum. Previous studies have shown that the requirement for turgor is conserved, although the osmoticum may differ. This information will likely facilitate the development of new approaches to managing the spread of pathogenic species by targeting the very processes that allow them to efficiently colonize new environments.
The genus Cryptoporus has historically been delimited primarily based on the distinctive veiled polypore morphology. Since its first report from North America, the genus has been regarded as monotypic, represented by Cryptoporus volvatus. Recently, two additional species were described from south and central China based on DNA sequence data. However, despite numerous records from Korea, Japan, and eastern China, no molecular phylogenetic studies have yet examined collections from these regions. The present study aims to clarify the species boundaries and geographic distribution of Cryptoporus in East Asia using integrative taxonomic approaches. Multilocus phylogenetic analyses were conducted using sequences from ITS, 28S, 18S, RPB2, and TEF1, together with species delimitation analyses. Our results demonstrate that East Asian collections represent a distinct lineage, herein described as a new species, Cryptoporus densiflorus. The new species is confirmed to occur in Korea and Japan and appears to be host-specific to Pinus densiflora. We reevaluate the geographic ranges of all currently accepted species and highlight that historical "C. volvatus" records in Asia include multiple taxa. These results indicate that Cryptoporus species exhibit strong geographic structuring and host association at the species level, thereby enhancing the taxonomic framework of the genus in East Asia.
Fungal infections caused by Candida albicans (C. albicans) pose a significant clinical challenge, especially due to the rise of multidrug-resistant strains. Antifungal peptides (AFPs) represent promising alternatives because of their high efficacy, low toxicity, and reduced risk of resistance. In this study, we integrated computational modeling and experimental validation to identify novel AFPs targeting the fungal membrane. Four candidate peptides-HIV-F9170 (GWEALKYLWNLLQYW), PinA (WRWWKWWK), PinB (WPTKWWK), and XYP1 (KIKWFKAMKSIAKFIAKDQLKKHL)-were evaluated using AlphaFold2 structure prediction, molecular dynamics (MD) simulations, and in vitro antifungal assays. MD simulations revealed that PinA and XYP1 exhibited strong binding and stable interactions with a model C. albicans membrane. Correspondingly, in vitro assays confirmed that these two peptides effectively inhibited fungal growth without cytotoxicity toward HEK-293 cells. Our findings provide a mechanistic basis for peptide antifungal activity and identify promising candidates for further therapeutic development.
Global environmental changes that characterize the Anthropocene, such as extreme weather events, increase in temperature, severe droughts, etc. are affecting soil fungi, the crucial roles they play in ecosystems, and therefore impacting our society. In the past two decades, many studies have investigated the effects of diverse environmental changes (i.e. global change drivers) in fungi, at the species, population, and community levels, in natural and managed ecosystems, as well as under laboratory conditions. In this review, we draw together these studies to examine overall impacts on fungi when exposed to global change drivers. In addition, we summarize how global change and impacts on fungi are affecting society through impacts on public health, food security, and biodiversity. In closing, we present ways forward for research in this area, as well as gaps in knowledge and emergent questions in the field of soil fungi in the Anthropocene.
The fungus Fusarium xyrophilum produces flower-like structures (i.e. pseudoflowers) composed entirely of mycelia on yellow-eyed grasses (Xyris spp.), including X. surinamensis in Guyana. Whether these pseudoflowers function as true mimics that attract insects for fungal propagule dispersal remains unknown. We evaluated the potential of F. xyrophilum to influence insect visitation patterns by (i) documenting insect visits to real flowers and pseudoflowers on X. surinamensis in the field in Guyana, (ii) assessing the presence of F. xyrophilum DNA on insect bodies, and (iii) analyzing volatile emissions from flowers and pseudoflowers. During our field work, a broad range of generalist insects (i.e. Vespidae, Formicidae, Salticidae, Acrididae, and Tetrigidae) visited the flowers and pseudoflowers, which notably shared some insect visitors (i.e. meadow katydids, carpenter ants, and geometer moths). Some of these insects were observed chewing on the flowers and pseudoflowers. Fusarium xyrophilum DNA was detected on the bodies of captured insects using conventional and quantitative polymerase chain reaction (PCR). Volatile profiles of pseudoflowers and flowers were largely similar, except for a sesquiterpene, putatively identified as α-gurjunene, which was emitted by pseudoflowers and pure cultures of F. xyrophilum but not Xyris flowers. These findings support the role of insect visitors as vectors of F. xyrophilum propagules and suggest that the fungus may attract and use generalist insects by exploiting their preexisting sensory biases for Xyris floral signals rather than true mimicry.
The genus Lanmaoa is a globally distributed, economically important group of mushroom-forming fungi in the family Boletaceae containing edible and psychoactive species. Despite the growing interest in its hallucinogenic properties, the fundamental systematics of this genus remains poorly resolved due to limited taxon sampling and limited genetic characterization. This study combines a comprehensive sampling of all Lanmaoa species, including 21 type specimens, with whole genome sequencing of 53 specimens. A phylogeny of Lanmaoa is generated based upon an alignment of 1515 single-copy orthologs and recovers full statistical support at all major nodes, resolving evolutionary relationships in the genus. Phylogenomic analysis of previously unsequenced type specimens supports several taxonomic changes, including six novel combinations and the discovery of four novel species, two of which are described here-Lanmaoa fallax, sp. nov. and Lanmaoa carbonilivor, sp. nov.-resulting in a total recognition of 17 species in the genus. Genome mining of Lanmaoa asiatica fails to detect canonical biosynthetic genes associated with psilocybin or ibotenic acid production, suggesting the presence of a novel psychoactive compound. This study establishes a comprehensive genomic foundation for Lanmaoa systematics, enabling future research to more robustly explore the evolutionary history and secondary chemistry of the genus.
The increasing frequency and severity of wildfires has necessitated assessing fire effects on soil systems. Variation in fuel loads and fire effects create landscape mosaics with distinct abiotic properties, a heterogeneity that has been dubbed pyrodiversity. Expanding on the pyrodiversity framework, the pyrodiversity-biodiversity hypothesis posits that pyrodiversity increases niche diversity, thereby promoting biodiversity. This hypothesis, however, has remained untested for soil microbes and microeukaryotes. We explored this hypothesis for soil fungal communities using pre- and post-fire data from three empirical fuel load manipulations and across a total of five different vegetation contexts. We first compared pre- and post-fire abiotic heterogeneity to test whether fuel load manipulations would lead to greater environmental heterogeneity, particularly in soil properties. We then tested whether such manipulations led to greater fungal biodiversity as measured by fungal richness, β-diversity, and community dispersion. Labile abiotic soil resource (e.g. plant available phosphorus and inorganic nitrogen) heterogeneity increased post-fire, but this effect depended on the experimental context. In contrast, we observed little evidence for pyrodiversity-associated increases in post-fire fungal richness or diversity; community dispersion increased only in the study with the most extreme fuel load manipulations. Although our analyses did not clearly answer whether pyrodiversity begets biodiversity, our results highlight the nuances of soil responses to fire. Pyrodiversity-biodiversity linkages appear to depend on the system and on the diversity metric: the hypothesis had no support based on fungal richness, but community dispersion provided some support, even if only in one experiment. Understanding system-specific responses may be particularly important as fires increase in systems where they have been suppressed or have been historically rare.
Sooty molds are produced by complex species mixtures of epifoliar, saprotrophic Pezizomycotina. While amber deposits dominate their fossil record, direct evidence on fossilized angiosperm leaves has remained elusive. This study describes seven distinct morphotypes of sooty molds, including moniliform hyphae, setae, pycnidia, and setose sporocarps, preserved in situ on angiosperm leaf cuticles from the Late Miocene Siwalik deposits (ca. 12-8 Ma) of Himachal Pradesh, India. These specimens provide the first direct evidence of multispecies sooty mold communities associated with angiosperm surfaces in the fossil record. This discovery confirms that the saprotrophic epiphyllous habit was well established in Siwalik Miocene forests, offering significant insights into the evolutionary history and paleoecological interactions of these fungal communities.
During a survey of plant root-associated microbiota in the Pennsylvania Chrome Serpentine Barrens, we identified two previously unrecognized fungal lineages from Panicum virgatum (switchgrass). Based on multigene phylogenetic analyses, along with morphological and ecological characterization, we propose the establishment of two new genera, Curvophialophora and Drepanoconia, together with their respective species, Curvophialophora panicicola, sp. nov. and Drepanoconia panicicola, sp. nov. in the family Magnaporthaceae. Curvophialophora is characterized by simple, undifferentiated, septate, hyaline conidiophores bearing terminal, strongly curved phialidic conidiogenous cells that produce aseptate, hyaline, ovoid to ellipsoidal conidia. Drepanoconia has complex, differentiated, straight, brown conidiophores with terminal, slightly curved phialidic conidiogenous cells producing aseptate, hyaline, strongly curved sickle-shaped conidia. The plant-fungal interaction experiment showed that both genera had significant inhibitory effects on Arabidopsis thaliana, indicating their phytotoxicity potential on plants. These findings expand our knowledge on the microbiota associated with switchgrass, an emerging biofuel crop, and contribute to the study of systematics, diversity, and function of Magnaporthaceae.
The fungi of California's xeric oak habitats and nearby areas of the Pacific states of American West have long been understudied due to their irregular fruiting patterns, low expected yields, and the challenges these pose for planning field work. This study is based on the authors' extensive research into this natural environment. This article is the first in a series of installments that will describe more than 24 new members of Cortinariaceae from this habitat. Here we describe nine new species in the genus Calonarius-Ca. ambrosius, Ca. ostentatus, Ca. laudabilis, Ca. pretiosus, Ca. praestabilis, Ca. eminens, Ca. involutus, Ca. inopinatus, and Ca. polychromus, all associated with Quercus, and in some cases with Notholithocarpus densiflorus species. Some species presented here appear to be limited to the xeric zone, whereas others are widespread but more common within it. A brief history of the study of Cortinarius s.l. in California is also presented.
The edible fungus Aspergillus oryzae is well known for its powerful ability to secrete numerous proteolytic enzymes, such as lipases, proteases, and amylases. However, the mechanisms that regulate the translation of these enzymes in A. oryzae remain poorly understood. In this study, we investigated the biological function of the translation initiation factor eIF3j in A. oryzae. Our findings revealed that deletion of the eIF3j gene significantly inhibited mycelial growth and conidiation compared with the control strain. Furthermore, the eIF3j mutant showed increased sensitivity to various stressors relative to the control strain. Interestingly, the eIF3j-deleted strain exhibited enhanced production of amylase and kojic acid when compared with the control strain. Additional quantitative proteomic analysis suggested that eIF3j was involved in RNA processing, ribosome biogenesis, and amino acid metabolism. Taken together, these findings underscore the multifaceted role of eIF3j in A. oryzae and its potential as a target for improving traits relevant to industrial applications.
Thelephora is a genus of ectomycorrhizal fungi with a global distribution. Recent taxonomic efforts have revealed new species and species complexes, often distinguished by subtle morphological differences. Here, we describe Thelephora renispora, a new species resembling T. versatilis and T. pseudoversatilis, and support its recognition as a distinct taxon based on morphological traits, phylogenetic analyses of nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS barcode) and D1-D2 domains of nuc 28S rDNA (28S), and statistical comparisons of its micromophology against closely related allies. New distribution records for T. pseudoversatilis are also provided. Additionally, we present phylogenetic informativeness analyses to assess the utility of ITS, 28S, and other markers previously used for the order Thelephorales. Our results identify the DNA-directed RNA polymerase II second-largest subunit (RPB2) as the most informative locus for delimiting species-level relationships in the Thelephorales order, whereas ITS contributes to solving interspecific relations within the Thelephora-Tomentella clade.
Taxonomic revisions in mycology often focus on well-studied fungal groups, yet obscure lineages such as Laboulbeniales (Pezizomycotina, Ascomycota) remain underexplored. Here, we address the taxonomy of Stigmatomyces sensu lato, a diverse genus of obligate biotrophic fungi parasitizing flies (Diptera). Previous work suggested that Stigmatomyces is paraphyletic, as the well-delineated genus Gloeandromyces was nested within the genus, which resulted in the reinstatement of Appendiculina and Fanniomyces. We reconstructed a molecular phylogeny of Stigmatomyces sensu lato using three genetic markers (18S and 28S nuc rDNA, TEF1α) and significantly expanded taxon sampling. Our results recovered six well-supported clades, including the three aforementioned genera, Stigmatomyces sensu stricto, and two unnamed lineages. Although the internal relationships among these clades remain unresolved, morphological distinctions further support their delimitation. We provide arguments both for and against splitting of the genus. A formal split of Stigmatomyces is not proposed due to still limited data and especially the lack of sequences from the type species, S. baeri. However, our findings are in support of the splitting of Stigmatomyces sensu lato in the future. Based on molecular phylogenetic, morphological, and ecological data, we also describe two new species from Chile, Appendiculina drosophilae-huillicheae on Drosophila huilliche (Drosophilidae) and Stigmatomyces beukii on Prosopantrum sp. (Cnemospathididae). Based on morphological, ecological, and/or molecular phylogenetic data, we propose four new combinations: Appendiculina dacina, A. majewskii, Bordea euconni, and Sphaleromyces montevideensis. Finally, we validly publish Stigmatomyces biformis and propose Stigmatomyces rakiurensis to accommodate S. australis M.B. Hughes, A. Weir, Gillen & W. Rossi, a later homonym of S. australis Thaxt.
