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Fossils can unveil a long-vanished combination of character states that inform inferences about the timing and patterns of diversification of modern fungi. By examining the well-preserved stacked chained vesicular conidiophores developed in clusters from the basal stroma, we describe a new taxon of fossil Zygosporiaceae with a combination of characters unknown among extant taxa on compressed serrated-margined dicot leaf (cf. Fagaceae) recovered from the Siwalik sediments (Late Miocene; ca. 12-8 Ma) of Himachal Pradesh, western Himalaya. Based upon conidiophore morphology, our Siwalik fungal remains, similar to Zygosporium Mont. (Zygosporiaceae: Xylariales: Sordariomycetes), are recognized as a new fossil species, Z. stromaticum Kundu & Khan, sp. nov. Zygosporium stromaticum is the only known fossil anamorphic fungus that occurs on plant cuticles and has a cluster of stacked chained vesicular conidiophores arising from a poorly preserved basal stroma formed by irregular, thick-walled cells. Its combination of morphological characteristics is unknown in extant fungal taxa, so Z. stromaticum likely represents a new anamorphic foliicolous fungus that may now be extinct. This unique evidence may be essential for the calibration of divergence time estimations of fungal lineages.

Pinibarrenia chlamydospora , sp. nov. isolated from the roots of highbush blueberry in the New Jersey Pine Barrens, is described and illustrated. Based on multigene phylogenetic analysis, as well as morphological and ecological characteristics, Pinibarreniales and Pinibarreniaceae are established to accommodate this novel lineage in Sordariomycetidae, Sordariomycetes. Pinibarreniales, Tracyllalales, and Vermiculariopsiellales are proposed to be included in the subclass Sordariomycetidae. Pinibarreniales likely have a wide distribution and forms association with Ericaceae plants that live in acidic and oligotrophic environments because its DNA barcode matches with environmental sequences from other independent ecological studies. The plant-fungal interaction experiment revealed negative impacts on Arabidopsis , indicating its pathogenicity. This uncovered new fungal lineage will contribute to a better understanding of the diversity and systematics of Sordariomycetes.

Evolutionary relationships between the morphological and ecological traits of fungi are poorly understood. The appendages of chasmothecia, which are sexual reproductive organs of Erysiphaceae, are considered to play a crucial role in the overwintering strategies of these fungi on host plants. Previous studies suggested that both the host type and appendage morphology evolved at the same nodes and transitioned from complex appendages on deciduous hosts to simple appendages on herb/evergreen hosts. However, the evolutionary dependence between host type and appendage morphology remains unproven owing to the limited species data used in analyses. To elucidate the evolutionary relationship between host type and appendage morphology, we used phylogenetic comparative methods (PCMs) to investigate the state transition, ancestral state, evolutionary dependence, and contingent evolution within Erysipheae, the largest and most diverse tribe in Erysiphaceae. Our PCMs, based on a comprehensive data set of Erysipheae, revealed that the most ancestral states were deciduous host types and complex appendages. From these ancestral states, convergent evolution toward the herb/evergreen host types and simple appendages occurred multiple times at the same nodes. For the first time in Erysiphaceae, we detected an evolutionary dependence between host type and appendage morphology. This is one of the few examples in which evolutionary dependence between host phenology and morphological traits in plant-parasitic fungi was demonstrated using PCMs. Appendage simplification on herb/evergreen hosts and complications on deciduous hosts can be reasonably explained by the functional advantages of each appendage type in different overwintering strategies. These expected appendage functions can explain approximately 90% of host type and appendage morphology combinations observed in the analyzed taxa. However, our results also highlighted the occurrence of evolutionary shifts that deviate from the expected advantages of each appendage morphology. These seemingly irrational shifts might be interpretable from the flexibility of overwintering strategies and quantification of appendage functions.

Fossil epifoliar fungi are valuable indicators of paleoenvironment and paleoecology. The Meliolaceae, members of which typically inhabit the surface of living plants as biotrophs or pathogens, is one of the largest groups of epifoliar fungi. In this study, we report a novel fossil species of Meliolinites Selkirk (fossil Meliolaceae), Meliolinites tengchongensis , on the lower epidermis of compressed fossil Rhodoleia (Hamamelidaceae) leaves from the Upper Pliocene Mangbang Formation of Tengchong, Yunnan, southwestern China. Meliolinites tengchongensis is characterized by web-like, superficial, brown to dark brown, septate, and branching mycelia bearing 2-celled appressoria and unicellular phialides. The fungal colonies also include ellipsoidal, 5-celled, 4-septate ascospores and dark brown perithecia with suborbicular outline and verrucose surface. The well-preserved vegetative and reproductive organs help us to explore the potential disease process of the new fossil species. Besides, the presence of fungal remains indicates that the fungal taxon might have maintained its host preference since at least the Late Pliocene. Furthermore, the occurrence of both fossil fungi and their host plants in Tengchong indicate a subtropical-tropical, warm, and humid climate during the Late Pliocene, whereas the distribution pattern of the fungi on the host leaves suggests that Rhodoleia may have been a part of the middle-upper canopies in the Tengchong Late Pliocene multilayered forest.

Proteomics has been used extensively in the field of mycology, mainly in trying to understand the complex network of protein-protein interactions that has been implicated in the molecular functions of fungi. It is also a useful tool to compare metabolic differences within a genus. Species of Pseudogymnoascus , a genus under the phyla Ascomycota, have been shown to play an important role in the soil environment. They have been found in both polar and temperate regions and are a known producer of many extracellular hydrolases that contribute to soil decomposition. Despite the apparent importance of Pseudogymnoascus spp. in the soil ecosystem, investigations into their molecular functions are still very limited. In the present study, proteomic characterization of six Pseudogymnoascus spp. isolated from three biogeographic regions (the Arctic, Antarctic, and temperate regions) was carried out using tandem mass spectrometry. Prior to proteomic analysis, the optimization for protein extraction was carried out. Trichloroacetic acid‑acetone‑phenol was found to be the best extraction method to be used for proteomic profiling of Pseudogymnoascus spp. The proteomic analysis identified 2003 proteins that were successfully mapped to the UniProtKB database. The identified proteins were clustered according to their biological processes and molecular functions. The shared proteins found in all Pseudogymnoascus spp. (1201 proteins) showed a significantly close relationship in their basic cellular functions, despite differences in morphological structures. Analysis of Pseudogymnoascus spp. proteome also identified proteins that were unique to each region. However, a high number of these proteins belonged to protein families of similar molecular functions, namely, transferases and hydrolases. Our proteomic data can be used as a reference for Pseudogymnoascus spp. across different global regions and a foundation for future soil ecosystem function research.

Members of the poorly investigated family Teloschistaceae in South America, mostly from Bolivia and Peru, were examined using molecular and morphological data here for the first time. In recent phylogenetic reclassifications of Teloschistaceae, South American representatives were poorly represented but shown to belong to subfamilies Teloschistoideae and Xanthorioideae. In this study, we expanded the sampling of South American taxa and investigated mainly the lobate, sublobate, and squamulose members of Caloplaca s.l., using morphological characters and a molecular phylogeny based on a combined three-locus data set (one mitochondrial and two nuclear loci). Building upon new phylogenies at the family and subfamily levels (Teloschistoideae), we propose here three new genera: Andina, Aridoplaca , and Cinnabaria , with the type species Andina citrinoides, Aridoplaca peltata , and Cinnabaria boliviana . We also propose to reduce Tarasginia to synonymy with Sirenophila and Tayloriellina to synonymy with Villophora and introduce three new combinations: Dufourea ottolangei, D. volkmarwirthii , and Villophora erythrosticta. Scutaria andina is reported as new to Bolivia. A critical revision of the subfamily Brownlielloideae confirmed recent findings that it is an artifactual taxon based on a "chimeric" data set, with the type genus being part of Teloschistoideae.

Three species of Xenosporium producing ellipsoidal or ovoid conidia were collected from decaying culms of Miscanthus floridulus submerged in freshwater streams of Alishan area, Chiayi County, Taiwan. Xenosporium formosiforme and X. parvulum are described and illustrated as new species. Xenosporium africanum is described in this paper as a new record for Taiwan. The phylogenetic relationships of Xenosporium species and related taxa were sought by comparing the sequences of their internal transcribed spacer (ITS) barcode, 28S rDNA, and RNA polymerase II second largest subunit ( RPB2 ) gene segments. Phylogenetic analysis of these Xenosporium species supports their placement in the Tubeufiaceae. A synopsis of Xenosporium species producing ellipsoidal or ovoid conidia is given. The criteria used to differentiate species are discussed.

Dictyochaeta (Chaetosphaeriaceae) is a phialidic dematiaceous hyphomycete with teleomorphs classified in Chaetosphaeria . It is associated with significant variability of asexual morphological traits, which led to its broad delimitation. In the present study, six loci: nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS barcode), nuc 18S rDNA (18S), nuc 28S rDNA (28S), DNA-directed RNA polymerase II second largest subunit gene ( RPB2 ), translation elongation factor 1-α ( TEF1-α ), and β-tubulin ( TUB2 ), along with comparative morphological and cultivation studies, are used to reevaluate the concept of Dictyochaeta and establish species boundaries. Based on revised species, morphological characteristics of conidia (shape, septation, absence or presence of setulae), collarettes (shape), and setae (presence or absence) and an extension of the conidiogenous cell proved to be important at the generic level. The dual DNA barcoding using ITS and TEF1-α , together with TUB2 , facilitated accurate identification of Dictyochaeta species. Thirteen species are accepted, of which seven are characterized in this study; an identification key is provided. It was revealed that D. fuegiana , the type species, is a complex of three distinct species including D. querna and the newly described D. stratosa . Besides, a new species, D. detriticola , and two new combinations, D. callimorpha and D. montana , are proposed. An epitype of D. montana is selected. Dictyochaeta includes saprobes on decaying wood, bark, woody fruits, and fallen leaves. Dictyochaeta is shown to be distantly related to the morphologically similar Codinaea , which is resolved as paraphyletic. Chaetosphaeria talbotii with a Dictyochaeta anamorph represents a novel lineage in the Chaetosphaeriaceae; it is segregated from Dictyochaeta , and a new genus Achrochaeta is proposed. Multigene phylogenetic analysis revealed that D. cylindrospora belongs to the Vermiculariopsiellales, and a new genus Tubulicolla is introduced.

Megacoelomyces (type species: Megacoelomyces sanchezii ), an ascomycete asexual morph infecting Myrcia fenzliana (Myrtaceae) from the Brazilian Cerrado, is described as a new genus in the Phaeosphaeriaceae (Pleosporales, Dothideomycetes, Ascomycota), based on multilocus phylogeny (three nuclear ribosomal DNA and two protein-coding genes) in addition to morphological (light and scanning electron microscopy) and ecological data.

This paper describes and illustrates a new species of Laboulbeniales (Ascomycota, Laboulbeniomycetes) recovered from Mastoptera guimaraesi bat flies (Diptera, Streblidae) in Ecuador and Panama. Bat fly-associated Laboulbeniales are still unexplored in the Neotropics, with only four described species of Gloeandromyces and one species of Nycteromyces known. Morphological characteristics and phylogenetic analyses support placement of the new taxon in Gloeandromyces and its recognition as an undescribed species. Gloeandromyces hilleri sp. nov. is easily recognized by 2-3 longitudinal rows of undulations at its perithecial venter. Phylogenetic reconstructions of the large subunit (LSU) ribosomal DNA and the translation elongation factor 1α ( TEF1 ) both resolve G. hilleri and G. nycteribiidarum as sister species. We discuss the utility of LSU and TEF1 as secondary barcodes in Laboulbeniomycetes taxonomy.

We are becoming increasingly aware of the dazzling diversity of fungi-new taxa are being discovered from poorly sampled habitats and integrative approaches point at (near-)cryptic species in many groups. The class Laboulbeniomycetes, which is composed of three orders (Herpomycetales, Laboulbeniales, Pyxidiophorales), is no exception. However, still, in the orders Herpomycetales and Laboulbeniales, species are predominantly described based on morphology alone. Here, we present a new species of Herpomyces from Argentina parasitic on Periplaneta fuliginosa , the smokybrown cockroach. Herpomyces spegazzinii , sp. nov., is characterized based on morphology and molecular data. Phylogenetic inference based on internal transcribed spacer (ITS, consisting of ITS1-5.8S-ITS2) barcode region supports the status of this fungus as a separate species, sister to the recently described H. shelfordellae .

Historically, thallus-forming Laboulbeniomycetes, including the orders Laboulbeniales and Herpomycetales, were set apart because of their distinctive morphology and ecology. Although some biologists correctly interpreted these arthropod ectoparasites as fungi, even ascomycetes, others thought they were worms, red algae, or members of taxa described especially for them. Speculation on the evolution of the group involving red algae, the morphology-based Floridean Hypothesis, persisted deep into the 20th century, in part because valid alternatives were not presented. Although the distinctive features of Laboulbeniales clearly set them apart from other fungi, the difficulty was in the absence of characters grouping them among the fungi. Thaxter considered the Laboulbeniales to be ascomycetes, but he avoided phylogenetic discussions involved in the Floridean Hypothesis all of his life. Eventually, developmental studies of the life history of Pyxidiophora species, hyphal perithecial ascomycetes with 2-celled ascospores, revealed characters connecting Laboulbeniales to other ascomycetes. The distinctive morphological features of Laboulbeniales (absence of mycelium, a thallus developed from 2-celled ascospores by cell divisions in several planes, arthropod parasitism) can be best understood by comparison with Pyxidiophora . The development of a 3-dimensional thallus composed of true parenchyma occurs not only in Laboulbeniales, but also in Pyxidiophora species. The life history of arthropod ectoparasitism of Laboulbeniales as well as mycoparasitism and phoretic dispersal by arthropods of Pyxidiophora species can be explained by Tranzschel's Law, originally applied to rust fungi. Molecular analyses including other arthropod-associated fungi have contributed to a better understanding of an enlarged class, Laboulbeniomycetes, which now includes a clade comprising Chantransiopsis, Tetrameronycha , and Subbaromyces . A two-locus phylogenetic tree highlights evolutionary and life history questions with regard to the placement of Herpomycetales as the first diverging lineage of the Laboulbeniomycetes. The sister group for all the Laboulbeniomycetes remains to be discovered.

Many animals have been shown to eat fungi and most truffle-like fungi depend on animals for spore dispersal via mycophagy. Although these interactions are widespread, they are understudied in many habitats. In this study, we show that bonobos ( Pan paniscus ) forage and feed on an undescribed truffle species in the rainforests of the Democratic Republic of Congo. Based on morphological and molecular assessment of collections, we show that the species eaten by bonobos is a previously undescribed taxon described here as Hysterangium bonobo . This species is known in the local Bantu language (Bongando) as simbokilo and is used for baiting traps to catch several species of small mammals. Our findings highlight the need for further research into mycophagy and systematics of sequestrate fungi in Africa.

Environmentally damaging invasive plants can also serve as reservoir hosts for agricultural pathogens. Microstegium vimineum is an invasive C4 annual grass that is present throughout the midwestern and eastern United States. It can reach high densities in disturbed areas such as crop-forest interfaces, which creates the potential for pathogen spillover from M. vimineum to agricultural crops and native plants. A previous study that surveyed disease on M. vimineum found a large-spored Bipolaris species that was widespread on M. vimineum and also isolated from co-occurring native grasses. Here, we report that the large-spored fungus isolated from M. vimineum and the native grass Elymus virginicus is Drechslera gigantea , based on comparison with published descriptions of morphological traits, and establish that D. gigantea is a pathogen of M. vimineum and E. virginicus . We review the phylogenetic placement and taxonomic history of D. gigantea and propose that it be reassigned to the genus Bipolaris as Bipolaris gigantea.

Truffles in the genus Tuber are hypogeus fungi that have a worldwide distribution. Despite this, knowledge about their diversity in the Middle East is very limited. In recent years, large quantities of truffles have been imported from Iran for being sold in Italy. While analyzing certain commercial batches of T. aestivum from Iran, we found some ascomata that resembled T. excavatum but had macro- and micromorphological features that were distinct from this species. They were subglobose, or depressed to slightly irregular, with a conspicuous basal cavity, grayish brown, brown, or pinkish gray, with a minutely papillose peridium. The gleba was pinkish gray in youth, brown at maturity, marbled with cream branched veins. Ascospores were broadly ellipsoid, with an irregular reticulum and distinctive long crests along the longitudinal axis, up to 9 µm high. Analysis of internal transcribed spacer (ITS) and large subunit (LSU) rDNA sequences showed that these specimens form a monphyletic and well-supported taxon within the Excavatum clade. Morphological and molecular analyses supported the proposal of the new species T. iranicum .

Truffles with distinct morphological and anatomical features were collected during a study of hypogeous fungi of semiarid sandy grasslands of the Great Hungarian Plain in Hungary, representing the westernmost localities of the Eurasian steppe belt. None of the ascomata were collected near ectomycorrhizal plant species, and none were identified as ectomycorrhizal during previous surveys in the collection area. We studied morphoanatomical characteristics of these truffles with light and scanning electron microscopy and investigated their phylogenetic positions based on analyses of different nuclear loci. The truffles were found to represent two novel lineages that grouped with the Marcelleina-Peziza gerardii clade of the Pezizaceae. One formed a distinct lineage, for which we propose a new genus Babosia with a new species Babosia variospora characterized by diverse spore ornamentation varying even within one ascus. The truffles in the other lineage clustered with the rarely collected American truffle Stouffera longii and share with it similar spore ornamentation and habitat features. However, our material differs from S. longii by geographic origin, the quick and strong coloration of the ascomata to dark gray at cut surface or bruised area, varying spore number in asci, and smaller spore size; thus, we describe it as a new species, Stouffera gilkeyae .

Cladosterigma clavariellum has been treated as a basidiomycete since its first description by Spegazzini in 1886 as Microcera clavariella . After further morphological studies, between 1919 and 2011, it remained among the basidiomycetes, most recently as incertae sedis in the order Cryptobasidiales. Our studies, based on light and scanning electron microscopy, supported by multilocus phylogenetic analyses-second-largest subunit of RNA polymerase II ( RPB2 ), translation elongation factor 1-alpha ( TEF1 ), small subunit (18S), large subunit (28S), and nuclear internal transcribed spacers (ITS1-5.8S-ITS2 = ITS) of the nuclear rDNA sequences, and mitochondrial rDNA small subunit (mtSSU)-finally determined the phylogenetic placement of Cladosterigma as the first nonlichenicolous mycoparasitic member of the Gomphillaceae within the Graphidales, an ascomycete order previously composed predominantly of lichen-forming fungi.

Four new species of Laboulbenia (Laboulbeniales, Ascomycota) occurring on Gerridae (Hemiptera, Insecta), a new host family, are described from six Central and South American countries: Bolivia, Brazil, Ecuador, Panama, Peru, and Venezuela. The new species are Laboulbenia brachymetrae, L. cylindrostethi, L. neogerris , and L. tachygerris .

Fly-speck fungi reproduce via thyriothecia that consist of sporogenous tissue appressed to cuticle surfaces of plant leaves and covered by a shield-like scutellum. Thyriothecial scutella likely evolved repeatedly in Dothideomycetes (Ascomycota), and their morphology varies by lineage. Fly-speck fungi have an exceptionally good fossil record that begins in the Mesozoic. The interpretation of scutellum characters in fossils may provide insights into origins of Dothideomycetes and help calibrate the timing of ascomycete evolution. From sediments of the Lower Cretaceous (125-112 Ma) Potomac Group of Virginia, from Dutch Gap Canal, lower Zone 1, we found scutella similar to those of extant Aulographaceae (Dothideomycetes), attached to a single piece of dispersed coniferous cuticle. We analyze hyphae and scutellum development among four extant Aulographaceae species for comparison with the fossil. The excellent preservation of fungi on the leaf cuticle surface allows us to infer a developmental sequence for the fossil. Scutellum development begins with coordinated growth of multiple neighboring generator hyphae and continues with hyphae producing two-dimensional pseudomonopodial, dichotomous, radial growth. Asci and ascospores were not found. We coded states for seven morphological characters using direct observations of the fossil and eight extant taxa, and using the literature for 28 others. We inferred a phylogeny using nuclear 18S and 28S rDNA of 36 extant taxa, 34 Dothideomycetes and two Arthoniomycetes. The phylogeny includes newly determined sequences from five species, two from Aulographaceae. With a branch-and-bound search, we inferred the most parsimonious placements of the fossil given the molecular tree topology. The parsimony analysis constrained by the rDNA phylogeny places the fossil taxon among stem lineages near Aulographaceae or among the known living members of Aulographaceae. We describe the fossil morphotype as Protographum luttrellii , gen. et sp. nov. The fossil provides the oldest evidence of morphological characters restricted among extant fungi to Aulographaceae.

The Diaporthales (Sordariomycetes) includes many important families of plant pathogenic fungi, such as the notorious Cryphonectriaceae. The aim of the present study was to reevaluate this family, along with other families in Diaporthales. Based on phylogenetic analyses using combined sequence data of the internal transcribed spacer (ITS) region, large subunit of the nrDNA (28S), and the translation elongation factor 1-alpha ( tef1-α ) and DNA-directed RNA polymerase II second largest subunit ( rpb2 ) genes, Cryphonectriaceae is separated into two subclades, comprising 21 genera and 55 species. Foliocryphiaceae, fam. nov., is morphologically and phylogenetically its closest relative but is distinct due to its phylogeny and dimorphic conidia. Mastigosporellaceae, fam. nov., is distinguished from other families in Diaporthales by owning apical conidial appendages. Neocryphonectria , gen. nov., within the family Foliocryphiaceae, with two species associated with Carpinus canker in China, is characterized by fusoid, aseptate macroconidia. Additionally, two new combinations are proposed, namely, Cryphonectria citrine , based on Chromendothia citrine , and Cytospora viridistroma , based on Endothia viridistoma . Based on results obtained in this study, 31 families are accepted into Diaporthales.

Podosphaera tridactyla (s. lat.) is a powdery mildew species occurring on a wide range of Prunus spp. almost worldwide. We have investigated the phylogeny of the Po. tridactyla complex, with special emphasis on potential aspects of cryptic speciation. The results suggested that Po. tridactyla represents a species complex consisting of at least 12 different species. Based on detailed morphological examinations and molecular sequence analyses, we propose dividing Po. tridactyla s. lat. into 10 species, encompassing 7 new species ( Po. ampla, Po. pruni-avium, Po. pruni-cerasoidis, Po. prunigena, Po. pruni-lusitanicae, Po. prunina , and Po. pruni-japonicae ) and 3 known species ( Po. longiseta, Po. salatai , and Po. tridactyla s. str.). Oidium passerinii on Pr. laurocerasus is confirmed as a synonym of Po. tridactyla s. str. Epitypes are designated for Po. tridactyla and Oidium passerinii.

Blueberry, an increasingly cultivated fruit crop in Portugal, is known to be susceptible to twig blight and dieback caused by species of Diaporthe . The diversity of Diaporthe species associated with symptomatic and asymptomatic Vaccinium corymbosum plants in Portugal was assessed. A multilocus sequence analysis of the rDNA internal transcribed spacer (ITS) region and the translation elongation factor 1-alpha ( tef1-α ), β-tubulin ( tub2 ), calmodulin ( cal ), and histone 3 ( his3 ) genes revealed the presence of Diaporthe foeniculina, Diaporthe rudis , and four new species, which are described as Diaporthe crousii, Diaporthe phillipsii, Diaporthe rossmaniae , and Diaporthe vacuae . These new species were characterized in terms of their morphology, mating strategies, and temperature growth requirements. In artificial inoculation trials of V. corymbosum cv. Bluecrop plants, all Diaporthe species caused minor symptoms. Further, no differences in aggressiveness were apparent between species. This study provides the first survey of Diaporthe species associated with blueberry twig blight and dieback in Portugal. It disclosed the occurrence of a diverse assemblage of Diaporthe species, whose status and impact as pathogens of blueberry is not yet fully understood.

The genus Magnaporthiopsis of Magnaporthaceae (Magnaporthales, Sordariomycetes, Ascomycota) contains species that are predominantly necrotrophic pathogens, often producing simple hyphopodia and dark, ectotrophic runner hyphae on plant roots and stems during colonization. Fungal isolates from turfgrass roots with dark and ectotrophic runner hyphae were examined and identified based on morphological, biological, and phylogenetic analyses. Maximum likelihood and Bayesian methods were implemented to obtain phylogenetic trees for partial sequences of the 18S nuc rDNA, ITS1-5.8S-ITS2 nuc rDNA internal transcribed spacer, and 28S nuc rDNA regions, and of the minichromosome maintenance complex 7 ( MCM7 ), largest subunit of RNA polymerase II ( RPB1 ), and translation elongation factor 1-alpha ( TEF1 ) genes. Our isolates consistently formed a distinct and highly supported clade within Magnaporthiopsis . These findings were reinforced by common and distinctive biological and morphological characters. Additionally, we conducted pathogenicity evaluations and demonstrated the ability of this fungus to colonize roots of ultradwarf bermudagrass, one of its native hosts, via ectotrophic, dark runner hyphae, causing disease symptoms including root discoloration and reduced root and shoot mass. Altogether, our discoveries enabled recognition and description of a new species, Magnaporthiopsis cynodontis , which has widespread distribution in the United States.

Corynespora cassiicola (Burk. & M.A. Curtis) C.T. Wei. is an anamorphic fungus that affects more than 530 plant species, including economically important crops. Several lineages of this pathogen have been recognized, but the classification of isolates into clades is time-consuming and still sometimes leads to unclear results. In this work, eight major phylogenetic clades (PhL1-PhL8) including 245 isolates of C. cassiicola from 44 plant species were established based on a Bayesian inference analysis of four combined C. cassiicola genomic loci retrieved from GenBank, i.e., rDNA internal transcribed spacer (ITS), actin-1 , ga4 , and caa5 . The existence of PhL1-PhL5 and PhL7 as clonal lineages was further confirmed through the analysis of full-genome single-nucleotide polymorphisms of 39 isolates. Haplotypes of the caa5 locus were PhL specific and encode isoforms of the LDB19 domain of a putative α-arrestin N-terminal-like protein. Evolution of the Caa5 arrestin is in correspondence with the PhLs. ga4 and caa5 PhL consensus sequences and a cleaved amplified polymorphic sequence (CAPS) procedure were generated based on the conserved nucleotide sequences and enzyme restriction patterns observed among isolates from the same lineage, respectively. The CAPS method was validated in silico , and its practical use allowed us to differentiate between tomato and papaya isolates, as well as to reveal the prevalence of PhL1 among isolates infecting soybean in Brazil. This novel approach could be useful in the efforts to control the diseases associated with C. cassiicola .

Acrospermum is a poorly known genus of epibiotic and saprophytic species with a subcosmopolitan distribution. Here, we investigate the intriguing relationship between Acrospermum and its host plants in the fern family Polypodiaceae, where it occurs upon approximately 45 neotropical species. We conducted phylogenetic analyses using an eight-marker comprehensive ascomycete data set comprising 719 species representing all major lineages along with 23 new Acrospermum specimens sampled from ferns. We ask whether fern-dwelling Acrospermum are monophyletic, whether epibiotic Acrospermum have evolved independently from saprophytic ancestors, and identify anamorphic phases by incorporating sequences for all suspected taxa. Our results corroborate the placement of Acrospermales within the Dothideomycetes with strong support. However, the order remains incertae sedis due to weak support along the branches subtending the clade that includes the Acrospermales plus Dyfrolomycetales. Our results show a strong phylogenetic pattern in lifestyles but do not clearly identify an ancestral life history state. The first divergence in Acrospermaceae splits fungicolous taxa from taxa that inhabit plants; saprophytes and anamorphic phases found on angiosperms occur in both clades. Fungicolous species are monophyletic, whereas species with an epibiotic or necrotic life history upon plants are nonmonophyletic due to the position of the saprophyte A. longisporium . Previously, all Acrospermum collected from ferns were identified as A. maxonii . Our results indicate that this is not monophyletic due to the inclusion of Gonatophragmium triuniae . Two species are described herein as A. gorditum , sp. nov., and A. leucocephalum , sp. nov. We find no instances of co-cladogenesis; however, our ability to detect this is limited by the lack of resolution in the A. maxonii clade. Rather, we see that that the distribution of epibiotic Acrospermum is explained by the overlap between the ecological niche of the Acrospermum species and its host.

Species of Septoria are commonly associated with leaf spot diseases of a broad range of plant hosts worldwide. During our investigation of fungi associated with leaf spot diseases in northern and northwestern Iran, several Septoria isolates were recovered from symptomatic leaves on different herbaceous and woody plants in the Asteraceae, Betulaceae, and Salicaceae families. These isolates were studied by applying a polyphasic approach including morphological and cultural data and a multigene phylogeny using a combined data set of partial sequences of the 28S nuc rRNA gene (large subunit [28S]), internal transcribed spacer regions and intervening 5.8S nuc rRNA gene (ITS) of the nuc rDNA operon, actin ( actA ), translation elongation factor 1-α ( tef1 ), calmodulin ( cmdA ), β-tubulin ( tub2 ), and DNA-directed RNA polymerase II second largest subunit ( rpb2 ). Four novel species are proposed, namely, Septoria eclipticola on Eclipta prostrata, Septoria firouraghina on Cirsium arvense, Septoria guilanensis on Populus deltoides , and Septoria taleshana on Alnus subcordata . All species are illustrated, and their morphology and phylogenetic relationships with other Septoria species are discussed.

The internal transcribed spacer (ITS) DNA marker is routinely used for fungal identification but gives a clear result for only three out of four powdery mildew samples. A search for new markers indicates that some genes offer enhanced identification in comparison with ITS. Others fail due to amplification and sequencing difficulties and lack of informative variability. Powdery mildews (Ascomycota, Erysiphales) are biotrophic, fungal plant pathogens that commonly occur worldwide on a wide range of host plants. They are unsightly and greatly reduce the vigor of their hosts and have major impacts on crop and other cultivated plants. Species within this order are straightforward to spot, but difficult to identify. A citizen science scheme was run in 2013-2016 in the UK to gather a wide array of samples on which identification methods could be developed. Current techniques for identification and phylogenetic reconstruction show scope for improvement. In this paper, we review genes used in other fungal groups for discrimination at species level. Working protocols for amplification and sequencing of seven genes ( actin, β-tubulin, calmodulin , Chs , elongation factor 1-α [ EF1-α ], Mcm7 , and Tsr1 ) are developed with varying success; Mcm7 proves to be the most useful at differentiation between closely related, phylogenetically young powdery mildew species for phylogenetic reconstruction when used separately and in tandem with ITS. We therefore propose this as the most appropriate candidate gene to be used commonly in powdery mildew diagnostics alongside the ITS; furthermore, this could be transferred to similarly troublesome fungal clades.

In this paper, species of the genus Morchella are investigated in China. Based on morphological characteristics and molecular phylogenetic analyses of the nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and the combined data set ITS + nuclear large subunit rDNA (28S) + the translation elongation factor 1-α ( TEF1 ) gene + RNA polymerase II first largest subunit ( RPB1 ) + RNA polymerase II second largest subunit ( RPB2 ), six new phylogenetic species are illustrated and described: M. clivicola, M. confusa, M. odonnellii, M. owneri, M. yangii , and M. yishuica . Furthermore, two new record species, M. dunensis and M. palazonii , which were only known in Europe, are now reported for the first time from Asia. New species of morels will provide additional information on species diversity and genetic resource candidates for improving the cultivation of this economically important fungus.

Species of Phaeochorella are biotrophic leaf parasites with a tropical distribution, traditionally accepted in the family Phyllachoraceae, Phyllachorales in classifications based on morphological characters. Phylogenetic evidence presented here resolves the relationship of Phaeochorella within the Sordariomycetes, based on a multilocus analysis of partial nuc rDNA large subunit (28S) and internal transcribed spacers (ITS1-5.8S-ITS2 = ITS), the DNA-directed RNA polymerase II second largest subunit ( RPB2 ), and the translation elongation factor 1-α ( TEF1-α ) gene. Phylogenetic analyses indicate that Phaeochorella belongs to the Diaporthales rather than the Phyllachorales. Phaeochorella parinarii , the type species of the genus, present on native hosts from the Brazilian Cerrado, forms a unique clade with a species of Phaeoappendicospora with high support. Thus, a new family, Phaeochorellaceae, Diaporthales, including both genera, is herein proposed. With the exception of P. parinarii and P. zonata , all other species in Phaeochorella ( P. artocarpi, P. ciliata, P. machaerii ) were excluded from the genus.

Micarea is a lichenized genus in the family Pilocarpaceae (Ascomycota). We studied the phylogeny and reassessed the current taxonomy of the M. prasina group. We focused especially on the taxonomic questions concerning the type species M. prasina and, furthermore, challenges concerning type specimens that are too old for successful DNA barcoding and molecular studies. The phylogeny was reconstructed using nuc rDNA internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS), mitochrondrial rDNA small subunit (mtSSU), and replication licensing factor MCM 7 gene from 31 species. Fifty-six new sequences were generated. The data were analyzed using maximum parsimony and maximum likelihood methods. The results revealed four undescribed, well-supported lineages. Three lineages represent new species described here as M. fallax, M. flavoleprosa , and M. pusilla . In addition, our results support the recognition of M. melanobola as a distinct species. Micarea fallax is characterized by a vivid to olive green thallus composed of aggregated granules and whitish or brownish apothecia sometimes with grayish tinge (Sedifolia-gray pigment). Micarea flavoleprosa has a thick, wide-spreading yellowish green, whitish green to olive green sorediate thallus and lacks the Sedifolia-gray pigmentation. The species is mostly anamorphic, developing apothecia rarely. Micarea melanobola is characterized by a pale to dark vivid green granular thallus and darkly pigmented apothecia (Sedifolia-gray). Micarea pusilla is characterized by a whitish green to olive green thinly granular or membranous thallus, numerous and very small whitish apothecia lacking the Sedifolia-gray pigment, and by the production of methoxymicareic acid. Micarea fallax, M. flavoleprosa , and M. melanobola produce micareic acid. The reliability of crystalline granules as a character for species delimitation was investigated and was highly informative for linking the old type specimen of M. prasina to fresh material.

Species of Apiognomonia are some of the most ubiquitous leaf-associated fungi of broad-leaved trees of the northern temperate zone. Especially widespread and diverse is Apiognomonia errabunda , found mostly on beech, oak, and linden. This species and its closest relatives are known for their complicated phylogenetic relationships that have caused considerable confusion in the past. In the present paper, we describe the results of a three-gene-based reconstruction of phylogenetic relationships between A. errabunda, A. veneta , and A. hystrix for 373 isolates. Using combined information from the nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS barcode) and intron regions of actin ( ACT ) and calmodulin ( CAL ) genes for 263 isolates, we confirmed the occurrence of host-associated lineages within A. errabunda . However, the clustering of isolates by host species was incomplete: some isolates occurred on the "wrong" hosts and a number of isolates carried mixed genetic profile, indicating substantial level of inter-host group recombination. In addition, a number of isolates were identified as putative, pending further verification, interspecies hybrids between the much more divergent A. errabunda and A. hystrix .

Recent collections of Tuber malacodermum from Spain, Corsica, and Mexico were compared in an integrative morphological and molecular phylogenetic framework, including study of type material. Phylogenetic analyses of nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) and nuc 28S rDNA (28S) regions showed that specimens from Spain and Corsica form a monophyletic group closely related to T. melosporum and T. rufum , whereas Mexican specimens form a clade within the T. lyonii species complex. Peridium and ascospore morphology contribute clear morphological distinctions among specimens from Spain and Corsica, Mexico, and the type specimen of T. malacodermum . Based on results of the morphological and molecular phylogenetic analyses, we assigned the specimens from Spain and Corsica to Tuber pustulatum , sp. nov., and the Mexican specimens to Tuber theleascum , sp. nov. We restrict T. malacodermum to the sole type material. Formal descriptions and illustrations of these taxa are provided.

Powdery mildew of Prunus spp. is a significant disease in most cherry growing regions of Washington, USA. Powdery mildews on Prunus virginiana and Pr. avium were previously assigned to Podosphaera clandestina s. lat. (= Po. oxyacanthae ) or Po. prunicola . In this report, we confirm the presence of two distinct Podosphaera species on these hosts. Phylogenetic analyses of nuc rDNA sequences from the internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS) and 28S subunit confirmed the presence of two distinct species. A morphological comparison with type material of Po. prunicola and additional collections demonstrated that the powdery mildew on Pr. virginiana (including var. demissa and var. melanocarpa ) is in fact Po. prunicola . The powdery mildew on Pr. avium is genetically, morphologically, and biologically distinct from Po. prunicola and is described here as the new species Po. cerasi . Cross-inoculation experiments confirmed that these two species are host specific. Podosphaera prunicola was unable to colonize Pr. avium , whereas Po. cerasi was unable to colonize Pr. virginiana . Morphological reexamination of numerous specimens identified as Po. prunicola on a broad range of Prunus species suggests that Po. prunicola is probably confined to species in Prunus subgen. Padus (= Prunus subgen. Cerasus sect. Laurocerasus , including sect. Padus ), with Pr. virginiana as the principal host. Podosphaera cerasi occurs on hosts in Prunus subgen. Cerasus , and our work confirms a newly described species of powdery mildew on Pr. avium . This work also includes the first documented and genetically proven European record of Po. prunicola on Pr. serotina and its widespread occurrence in the United States.

Lichen-inhabiting fungi are highly specialized mycoparasites, commensals or rarely saprotrophs, that are common components of almost every ecosystem, where they develop obligate associations with lichens. Their relevance, however, contrasts with the relatively small number of these fungi described so far. Recent estimates and ongoing studies indicate that a significant fraction of their diversity remains undiscovered and may be expected in tropical regions, in particular in hyperdiverse fog-exposed montane forests. Here, we introduce the new genus Zhurbenkoa , from South America and Europe, for three lichenicolous fungi growing on thalli of the widespread lichen genus Cladonia (Lecanorales). Phylogenetic analyses based on combined sequence data of mt and nuc rDNA obtained from Andean populations (Bolivia) placed Zhurbenkoa as a member of Malmideaceae, a recently introduced family of lichen-forming fungi in the class Lecanoromycetes. Zhurbenkoa is closely related to the genera Savoronala and Sprucidea . The new genus is characterized by the development of grayish brown to almost black apothecia lacking an evident margin, an epihymenium interspersed with crystals (often seen as pruina), a strongly conglutinated hymenium made of noncapitate and sparsely branched paraphyses, a colorless exciple composed of radially arranged hyphae, a Lecanora / Micarea -like ascus type, and aseptate or 1-septate ellipsoidal colorless ascospores. Zhurbenkoa includes two Neotropical ( Z. cladoniarum, Z. latispora ) and one widespread ( Z. epicladonia ) species. The lichenicolous trophic mode is documented for the first time in the Malmideaceae, which until now included only lichen-forming associations between fungi and green algae.

True morels ( Morchella spp.) are a group of edible fungi that are in high demand worldwide. However, this group of fungi remains poorly understood in terms of its genetic structure, life cycle, and mating system. In the present study, we cloned the MAT locus in Morchella sp. Mes- 20 using long-range polymerase chain reaction (PCR) amplification. Our results showed that the MAT1-2 idiomorph in the single-ascospore isolate YAASMCB-3 was 7.5 kb in length, harboring a single MAT1-2-1 gene, whereas the MAT1-1 idiomorph in the isolate YAASMCB-15 was 7.8 kb, carrying both MAT1-1-1 and MAT1-1-10 genes. Mating-type diagnostic assays of single-ascospore populations confirmed that Mes- 20 is heterothallic. In addition, 42 collections belonging to 17 morel species (7 black morel species [Elata clade], 9 yellow morel species [Esculenta clade], and 1 species from the Rufobrunnea clade) were used to clone and characterize the MAT1-1-1 and MAT1-2-1 genes for development of the mating gene genealogies. In concordance with the multilocus phylogenetic trees, collections for the respective species were readily distinguished by well-supported lineages in mating gene genealogies. The topologies were consistent within the Elata clade, whereas the branching order and sister-group relationships slightly varied within the Esculenta clade. Our results show that species in the genus Morchella could be delimited by mating gene genealogies.

Three new species, Nycteromyces orientalis, Dimeromyces capensis, and D. streblidarum, belonging to the Laboulbeniales (Fungi, Ascomycota) and collected on bat flies (Diptera, Streblidae), are described and illustrated. The new species of Nycteromyces is the second described in this rarely collected genus, with the type species, N. streblidinus, previously collected only three times from Venezuela, Panama, and Honduras. The material for our new species comes from Australia, India, Philippines, and Tanzania, and morphological characters of these new collections require emendation of the original generic diagnosis. The distribution of N. streblidinus is also extended with additional collections from Costa Rica and Peru. We describe two new species of Dimeromyces from streblid hosts, the first report of any members of this large, diverse genus on this host family. Newly described taxa are compared with known species and the genera Nycteromyces and Dimeromyces discussed in relation to other known Laboulbeniales taxa.

Gyromitra species are known as "false morels" and produce cerebriform or discoid ascocarps. The genus is widely distributed in the Northern Hemisphere, and some species are poisonous. Infrageneric phylogenies based on 28S nuc rDNA (28S) are available, but molecular analyses derived from other genes are lacking. In this study, Gyromitra specimens deposited in HMAS were reexamined. Phylogenies inferred from 28S, nuc rDNA internal transcribed spacers (ITS1-5.8S-ITS2 = ITS), and the translation elongation factor 1-α (TEF1) gene were obtained, and four clades were revealed among 13 species. Two new cryptic species of Gyromitra, G. pseudogigas, sp. nov. and G. tianshanensis, sp. nov., are described. Gyromitra pseudogigas has saddle-shaped apothecia similar to those of G. infula and is sister to G. gigas in the phylogeny. Gyromitra tianshanensis clusters with G. infula and G. xinjiangensis but differs in morphology and sequence data.

Rock-inhabiting fungi (RIF) are adapted to thrive in oligotrophic environments and to survive under conditions of abiotic stress. Under these circumstances, they form biocoenoses with other tolerant organisms, such as lichens, or with less specific phototrophic consortia of aerial algae or cyanobacteria. RIF are phylogenetically diverse, and their plastic morphological characters hamper the straightforward species delimitation of many taxa. Here, we present a phylogenetic study of two RIF genera, Lichenothelia and Saxomyces. Representatives of both genera inhabit rather similar niches on rocks, but their phylogenetic relationships are unknown so far. The cosmopolitan genus Lichenothelia is recognized by characters of fertile ascomata and includes species with different life strategies. In contrast, Saxomyces species were described exclusively by mycelial characters found in cultured isolates from rock samples collected at high alpine elevations. Here, we use an extended taxon sampling of Dothideomycetes to study the phylogenetic relationships of both Lichenothelia and Saxomyces. We consider environmental samples, type species, and cultured isolates of both genera and demonstrate their paraphyly, as well as the occurrence of teleomorphs in Saxomyces. We applied three species delimitation methods to improve species recognition based on molecular data. We show the distinctiveness of the two main lineages of Lichenothelia (Lichenotheliales s. str.) and Saxomyces and discuss differences in species delimitation depending on molecular markers or methods. We revise the taxonomy of the two genera and describe three new taxa, Lichenothelia papilliformis, L. muriformis, and Saxomyces americanus, and the teleomorph of S. penninicus.

Morphological and molecular phylogenetic studies of true morels (Morchella) in North America, the Dominican Republic, Venezuela, Ecuador, and Peru led to the discovery of four undescribed species of Morchella. Two new species in the Elata clade, one from the Dominican Republic, initially distinguished by the informal designation Mel-18, and a newly discovered sister species from northern Arizona, are now recognized. Mel-18 is described as a novel phylogenetically distinct species, M. hispaniolensis. Its sister species from Arizona is described as M. kaibabensis, also recovered as an endophyte of Rocky Mountain juniper. Two additional species in the Esculenta clade, M. peruviana discovered in Peru and M. gracilis (previously reported as Mes-14) from the Dominican Republic, Venezuela, and Ecuador, are described as new. We also demonstrate that scanning electron microscopy (SEM) imaging of ascospores using rehydration/dehydration/critical point drying preparation techniques provides for enhanced resolution of spore wall surfaces, thereby increasing the number of morphological traits available to assess differences among otherwise closely related species.

Following the abandonment of dual nomenclature and the implementation of single-name nomenclature for pleomorphic fungi, Coryneum was considered to have priority over Pseudovalsa and was recommended for use. Currently, Coryneum is the only genus in the family Coryneaceae (Diaporthales). However, DNA sequence data are lacking for most Coryneum species, and no detailed phylogenetic analyses of the genus are yet available. In the present study, fresh Coryneum samples were collected from chestnut (Castanea) and oak (Quercus) trees in China and morphologically compared with accepted Coryneum species. Based on morphological characteristics, they were identified as one known species, Coryneum castaneicola, and three novel species described here as C. gigasporum, C. sinense, and C. suttonii. Conidial dimensions and host association were considered major characters for species distinction. The previously unknown sexual morph of C. castaneicola is reported and described. A phylogenetic analysis of nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) and large subunit (28S) sequence data of a representative matrix of Diaporthales confirmed Coryneaceae to represent a monophyletic clade. A phylogenetic analysis of a combined sequence matrix containing the ITS-28S rDNA, the translation elongation factor 1-α (TEF1α), and the second largest subunit of the RNA polymerase II (RPB2) of the four Chinese and four additional European Coryneum species was performed, confirming the distinctness of these novel species.

The ascomycete genus Huntiella (Microascales) has a cosmopolitan distribution and occurs on a wide range of woody plants. Little is known regarding the identity, diversity, origin, or impact of these fungi in China. Recently, isolates of Huntiella spp. were collected from stumps of freshly felled trees or wounds on plantation-grown Eucalyptus in Guangdong, Guangxi, Fujian, and Hainan provinces of southern China. Additional isolates were obtained from stumps of Acacia confusa near Eucalyptus plantations in Hainan Province. The aim of this study was to identify these Huntiella species and to test their pathogenicity on Eucalyptus seedlings. Morphology and multigene phylogenies of the nuclear rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) region and partial β-tubulin (BT1) and translation elongation factor 1α (TEF1α) genes revealed nine previously unknown Huntiella species, eight from Eucalyptus and one from A. confusa. The mating types of these species were determined, showing that seven are heterothallic, one is homothallic, and one is unisexual (MAT1-2-1 gene). Pathogenicity tests showed that the nine Huntiella species can produce lesions on Eucalyptus seedlings, larger than wounds caused by controls on these plants. This study provides a basic understanding of the distribution, diversity, and pathogenicity of Huntiella species in southern China.

Sooty blotch and flyspeck (SBFS) fungi infect the cuticle of fruit, including apple fruit, and produce pigmented colonies. A new member of this fungal complex in the genus Peltaster is described on the basis of molecular and morphological evidence. The SBFS complex is a diverse group of ectophytic fungi that reside primarily within the order Capnodiales. Sooty blotch and flyspeck isolates from apple orchards in the central United States were subjected to parsimony and Bayesian analyses based on the internal transcribed spacer regions of nuc rDNA, the partial translation elongation factor 1-α gene, and the partial mitochondrial small subunit rRNA gene. Phylogenetic analysis delineated a new species, Peltaster gemmifer, from P. cerophilus and P. fructicola. Peltaster gemmifer conidiophores bear primary conidia that produce secondary conidia either through budding or through microcyclic conidiation; these were not seen in cultures of P. cerophilus and P. fructicola. On cellulose membrane that was placed on water agar amended with apple juice, P. gemmifer produced brown to black pycnothyria in a superficial brownish mycelial mat, similar to the colonies produced on apple fruit. Findings from the present study add to the >80 named and putative SBFS species so far described worldwide.

Lophodermium is a large fungal genus consisting of over 100 named species, with ca. 38 of these commonly found as endophytes of pine needles. In this study, we use both morphological and sequencing data to describe a new Lophodermium species associated with haploxylon pines from the Pacific Northwest. This new species resembled the morphology of L. nitens, another commonly occurring species from the same geographic regions and host species. They both present dark subcuticular ascocarps without lips. However, the upper walls of their ascocarps are different, as the new species forms an inward V-shaped folding, not present in L. nitens. Phylogenies using nuc rDNA internal transcribed spacer barcodes (ITS1-5.8S-ITS2 = ITS), partial D1-D2 domains of nuc rDNA 28S, and partial sequences of the nuc actin gene confirmed that this species represents a unique lineage not closely related to L. nitens. We discuss the current state of the phylogeny in light of all currently available sequences from pine-associated Lophodermium species.

Four new species, Tuber crassitunicatum, T. lishanense, T. piceatum, and T. wanglangense, are described and illustrated based on both morphological and molecular data. Morphologically, T. crassitunicatum is diagnosed by its brown ascomata and ellipsoidal ascospores ornamented by crowded spiny reticulum with more than 10 meshes across the spore width. Tuber lishanense can be recognized by its whitish to pale brown ascomata with a basal cavity, and very broadly ellipsoidal to subglobose ascospores ornamented by densely isolated spines. Tuber piceatum differs in its pale white ascomata and ellipsoidal ascospores with isolated spines, whereas T. wanglangense is characterized by its broad ellipsoidal ascospores with short spines that are connected by lower ridges when examined under scanning electron microscope (SEM). Phylogenetic analyses inferred the four new species in the Rufum group. Each species had less than 96% similarity in the internal transcribed spacer (ITS1-5.8S-ITS2) nuc rDNA ITS sequence to other Tuber species and represented a unique terminal clade in the phylogenetic tree. Our research did not confirm the occurrence in China of the European T. rufum and T. nitidum and the North American T. lyonii, although they are often reported in the literature on Chinese Tuber species.

Two new genera, Rhamphoriopsis and Xylolentia, are described for lignicolous perithecial ascomycetes occurring in terrestrial habitats. Fresh material, living cultures, morphology, and DNA sequence data (nuc rDNA internal transcribed spacers [ITS1-5.8S-ITS2 = ITS], 18S and 28S genes, and second largest subunit of RNA polymerase II = RPB2) of these taxa and morphologically similar fungi were studied to determine their relationships. A monophyletic clade including Rhamphoria, Rhodoveronaea, a dematiaceous hyphomycete Linkosia multiseptum, and the two new genera was recovered in the Sordariomycetes based on the 18S-28S-RPB2 data set. It is introduced as the family Rhamphoriaceae and strongly supported by Bayesian and maximum likelihood methods. Its members are characterized by perithecial ascomata with a cylindrical or rostrate neck, the absence of stromatic tissue or clypeus, similar anatomy of two-layered ascomatal walls, cylindrical paraphyses, unitunicate asci with a distinct, nonamyloid apical annulus, and dictyoseptate or transversely septate, hyaline or brown ascospores. The mode of conidiogenesis is holoblastic, predominantly on polyblastic-denticulate conidiogenous cells. The Phaeoisaria-like anamorph has been linked to Rhamphoria and Rhamphoriopsis, whereas conidia and conidiophores of Idriella-like synanamorph were formed in vitro in two species of Rhamphoria. The Veronaea-like anamorph is associated with Rhodoveronaea. The anamorph of Xylolentia is a dematiaceous hyphomycete with conidiogenous cells with sympodially extending rachis. A key to members of the family is provided. The classification and nature of species boundaries in Rhamphoria are discussed, and diagnostic characters such as ascospore shape, number of transverse and longitudinal septa, a degree of constriction at the septa, and ability to produce ascoconidia are evaluated.

Apiosphaeria guaranitica, the causal agent of brown crust disease of several bignoniaceous hosts, among them Handroanthus and Tabebuia species, has been traditionally placed in Phyllachoraceae, based exclusively on morphological studies, without supporting molecular evidence. Here, we provide molecular data for the link between sexual and asexual states of the fungus and elucidate the phylogeny of A. guaranitica. The multilocus phylogenetic analyses employed sequences from the 18S subunit (18S), 28S subunit (28S), and nuclear internal transcribed spacers (ITS1-5.8S-ITS2 = ITS) of the nuc rDNA, second-largest subunit of RNA polymerase II (RPB2), and translation elongation factor 1-α (TEF1) genetic loci. Estimates of the divergence time of this lineage were supported by fossil calibration (FC) and secondary calibration (SC) strategies. Our results indicate a natural placement of Apiosphaeria within Diaporthaceae (Diaporthales), where it represents an ancient lineage of the crown group of Diaporthaceae, diverging during the late Paleocene at 61.15 (FC) and 60.63 (SC) million years ago. This divergence time estimate within Diaporthales is based on Spataporthe taylori, a diaporthaceous fossil.

A multigene phylogenetic analysis of Genabea, Myrmecocystis, and related genera in Pyronemataceae was conducted to establish genetically supported generic limits. The nuc rDNA 28S gene, translation elongation factor 1-α gene (TEF1), and RNA polymerase II second-largest subunit gene (RPB2) significantly supported the monophyly of several distinct genera of hypogeous Pyronemataceae. The genetic identity of Genabea fragilis is established, and the new species Genabea hyalospora, G. urbana, Myrmecocystis mediterranea, and M. microspora are proposed to accommodate undescribed lineages. Two rare species, M. sphaerospora and M. spinospora, are tentatively identified based on collections that are morphologically similar to the original descriptions. The genus Genea was found to be monophyletic, except for G. cazaresii, which is nested among epigeous species of Humaria. The new combination Humaria cazaresii is proposed to accommodate this species.

Fungal root endophytes, including the common group of dark septate endophytes (DSEs), represent different taxonomic groups and potentially diverse life strategies. In this study, we investigated two unidentified helotialean lineages found previously in a study of DSE fungi of semiarid grasslands, from several other sites, and collected recently from a pezizalean truffle ascoma and eggs of the cereal cyst nematode Heterodera filipjevi. The taxonomic positions and phylogenetic relationships of 21 isolates with different hosts and geographic origins were studied in detail. Four loci, namely, nuc rDNA ITS1-5.8S-ITS2 (internal transcribed spacer [ITS]), partial 28S nuc rDNA (28S), partial 18S nuc rDNA (18S), and partial RNA polymerase II second-largest subunit (RPB2), were amplified and sequenced for molecular phylogenetic analyses. Analyses of similar ITS sequences from public databases revealed two globally distributed lineages detected in several biomes from different geographic regions. The host interaction of isolates from nematodes was examined using in vitro bioassays, which revealed that the fungi could penetrate nematode cysts and colonize eggs of H. filipjevi, confirming observations from field-collected samples. This is the first report of a DSE, and we are not aware of other helotialean fungal species colonizing the eggs of a plant-parasitic nematode. Neither conidiomata and conidia nor ascomata formation was detected in any of the isolates. Based on molecular phylogenetic analyses, these isolates represent a distinct lineage within the Helotiales in the Hyaloscyphaceae. For this lineage, we propose here the new genus Polyphilus represented by two new species, P. sieberi and P. frankenii.

Collections of a species referred to Sarcosomataceae (Pezizomycetes) from eastern North America were studied both morphologically and using nuc rDNA internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2 = ITS) and approximately 800 bp from the 5' region of the nuc 28S rDNA (28S) to construct a phylogeny. The analyses indicate that these collections are Donadinia seaveri, a species previously known only from Bermuda. Because the associated tree, Juniperus bermudiana, has declined as a result of insect attack, it was thought that D. seaveri might be extinct. This work indicates that it is not extinct but is present in eastern North America. The species is described, new distributional records are given, and its association with the genus Juniperus is discussed.

Four new species of Prolixandromyces (Laboulbeniales, Ascomycota) found on Veliidae (Heteroptera) from Bolivia, Brazil, Peru, and Venezuela are described and illustrated. These four species, Prolixandromyces anseris, P. tritici, P. blackwelliae, and P. bromelicola, represent the first records of this genus from South America, and their discovery requires emendation of the original generic circumscription. The newly described fungi are compared with known species, and a new key to identification is provided.