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Chanterelles (genus Cantharellus) are ectomycorrhizal fungi associated with a broad host range of trees. We explored Cantharellus diversity in mixed oak forests across 12 counties in Virginia and constructed a multilocus phylogeny including large ribosomal subunit (28S), transcription elongation factor 1-alpha (tef1), and RNA polymerase II (rpb2) sequences from all known temperate species of Cantharellus. Habitat and morphological attributes, including spore size, shape, and color, were compared with described species. In total, 104 sequences from 31 specimen vouchers were generated and 11 taxa identified, including one new species, C. sabuletorum from sandy Quercus-Pinus forests of Chesapeake Bay and north-central Florida. This study also provides evidence of genetic and color variation within C. cinnabarinus and expands the known distribution of C. altipes, C. appalachiensis, C. flavolateritius, C. lateritius, C. lewisii, C. minor, C. tenuithrix complex, C. vicinus, and C. velutinus in the eastern United States. 

Abstract PremiseThe agaricomycete order Cantharellales contains approximately 1000 species of fungi characterized by diverse morphological forms, ecological guilds, and nutritional modes. Examples include coralloid lichens that form symbioses with unicellular green algae, bulbil‐forming lichenicolous species, corticioid free‐living fungi that degrade dead sources of organic carbon, pathogens that cause plant disease, orchid root endosymbionts, and ectomycorrhizal fungi including popular edible mushrooms. However, evolutionary relationships in the Cantharellales remain poorly understood due to conflicting estimates based on ribosomal DNA loci. MethodsWe constructed a five‐gene phylogeny of the Cantharellales using data from 301 specimens to evaluate family‐level relationships. We used penalized likelihood to estimate divergence times and ancestral state reconstruction to test the hypothesis of multiple independent origins of biotrophic ecologies in the order and whether those transitions are younger than the divergence times of associated plant or lichen hosts. ResultsFour monophyletic families were recovered with strong support: Botryobasidiaceae, Ceratobasidiaceae, Hydnaceae s.l., and Tulasnellaceae, with Hydnaceae containing the greatest species richness and morphological diversity. Our results suggest the Cantharellales diverged during the Carboniferous period with subsequent diversification following the Permian‐Triassic extinction. Ancestral state reconstruction supports a saprotrophic most recent common ancestor with at least three transitions to an ectomycorrhizal ecology, multiple transitions to a lichenicolous habit with one or more subsequent transitions to mutualistic nutritional modes, four transitions to an orchid mycorrhizal ecology, and two transitions to a lichenized lifestyle. ConclusionsThis study represents the first comprehensive examination of the evolution of form and function across this ecologically and morphologically diverse order of fungi. 

The Ordway-Swisher Biological Station (OSBS) is a 38-km2 reserve owned by the University of Florida and is part of the National Ecological Observatory Network (NEON). The reserve contains several iconic Florida habitats, such as sandhill, mesic hammock, and scrubby flatwoods. While plants and animals have been extensively studied at OSBS, the fungi remain poorly known. Fungal inventories are critical to increase knowledge of both fungal diversity and species ranges, and thus to provide foundational data for a wide array of applications in ecology and resource management. Here, we present the results of a nine-year effort to collect, preserve, and DNA barcode the macrofungi at OSBS. This effort generated >1200 vouchered specimens and 984 ITS rDNA sequences, representing more than 546 species. Our sampling was dominated by Basidiomycota and revealed a high diversity of symbiotic ectomycorrhizal fungi, particularly species of Amanita, Cortinarius, and Russula. Sampling curves and both Chao1 and Jacknife1 richness estimators suggest that our DNA barcoding efforts captured only about half of the macrofungi species and that a more complete inventory would detect 897–1177 macrofungi species at OSBS. Our sampling found more species of macrofungi at OSBS than the known number of vertebrate animal species at the reserve and our estimates also suggest that there are likely more macrofungi species than plant species at OSBS. This study is the first comprehensive macrofungi inventory within a NEON site and highlights the importance of long-term monitoring to provide novel data on fungal diversity, community structure, conservation, biogeography, and taxonomy. 

Macroecological rules have been developed for plants and animals that describe large-scale distributional patterns and attempt to explain the underlying physiological and ecological processes behind them. Similarly, microorganisms exhibit patterns in relative abundance, distribution, diversity, and traits across space and time, yet it remains unclear the extent to which microorganisms follow macroecological rules initially developed for macroorganisms. Additionally, the usefulness of these rules as a null hypothesis when surveying microorganisms has yet to be fully evaluated. With rapid advancements in sequencing technology, we have seen a recent increase in microbial studies that utilize macroecological frameworks. Here, we review and synthesize these macroecological microbial studies with two main objectives: (1) to determine to what extent macroecological rules explain the distribution of host-associated and free-living microorganisms, and (2) to understand which environmental factors and stochastic processes may explain these patterns among microbial clades (archaea, bacteria, fungi, and protists) and habitats (host-associated and free living; terrestrial and aquatic). Overall, 78% of microbial macroecology studies focused on free living, aquatic organisms. In addition, most studies examined macroecological rules at the community level with only 35% of studies surveying organismal patterns across space. At the community level microorganisms often tracked patterns of macroorganisms for island biogeography (74% confirm) but rarely followed Latitudinal Diversity Gradients (LDGs) of macroorganisms (only 32% confirm). However, when microorganisms and macroorganisms shared the same macroecological patterns, underlying environmental drivers (e.g., temperature) were the same. Because we found a lack of studies for many microbial groups and habitats, we conclude our review by outlining several outstanding questions and creating recommendations for future studies in microbial ecology. 

A systematic reevaluation of North American pyrophilous or “burn-loving” species of Pholiota is presented based on molecular and morphological examination of type and historical collections. Confusion surrounds application of the names P. brunnescens, P. carbonaria, P. castanea, P. fulvozonata, P. highlandensis, P. molesta, and P. subsaponacea, with multiple names applied to a single species and multiple species described more than once. Molecular annotations using nuc rDNA ITS1-5.8S-ITS2 (internal transcribed spacer [ITS] barcode) and RPB2 (RNA polymerase II second largest subunit) are used to aid in application of these names in a phylogenetic context. Based on ITS molecular annotations of 13 types, the following heterotypic synonymies are proposed: P. highlandensis (syn. P. carbonaria and P. fulvozonata); P. molesta (syn. P. subsaponacea); and P. brunnescens (syn. P. luteobadia). In addition, we observed that the species P. castanea, known previously only from the type collection in Tennessee, is found commonly on burned sites near the Gulf Coast and other southeast regions of the United States. Overall, the pyrophilous trait is evolutionarily derived in Pholiota. Endophytic and endolichenic stages were deduced for P. highlandensis, the most widely distributed of the pyrophilous Pholiota. As a result, we introduce the “body snatchers” hypothesis that explains the maintenance of some pyrophilous fungi in ecosystems as endophytes and/or endolichenic fungi. Photographs, taxonomic descriptions, and a dichotomous key to pyrophilous species of Pholiota that occur in North America are presented. 

Abstract Multi‐locus sequence data are widely used in fungal systematic and taxonomic studies to delimit species and infer evolutionary relationships. We developed and assessed the efficacy of a multi‐locus pooled sequencing method using PacBio long‐read high‐throughput sequencing. Samples included fresh and dried voucher specimens, cultures and archival DNA extracts of Agaricomycetes with an emphasis on the order Cantharellales. Of the 283 specimens sequenced, 93.6% successfully amplified at one or more loci with a mean of 3.3 loci amplified. Our method recovered multiple sequence variants representing alleles of rDNA loci and single copy protein‐coding genesrpb1,rpb2 andtef1. Within‐sample genetic variation differed by locus and taxonomic group, with the greatest genetic divergence observed among sequence variants ofrpb2 andtef1 from corticioid Cantharellales. Our method is a cost‐effective approach for generating accurate multi‐locus sequence data coupled with recovery of alleles from polymorphic samples and multi‐organism specimens. These results have important implications for understanding intra‐individual genomic variation among genetic loci commonly used in species delimitation of fungi.