Variation in the SSUrDNA of the Genus Protostelium Leads to a New Phylogenetic Understanding of the Genus and of the Species Concept for Protostelium mycophaga (Protosteliida, Amoebozoa).

Members of the genus Protostelium (including P. mycophaga, P. nocturnum, and P. okumukumu) are protosteloid amoebae commonly found in terrestrial habitats on dead plant matter. They, along with the closely allied nominal genus Planoprotostelium, containing the single species Pl. aurantium, all have an amoeboid trophic stage with acutely pointed subpseudopodia and orange lipid droplets in the granuloplasm. These amoebae form stalked fruiting bodies topped with a single, usually deciduous spore. The species are identified based on their fruiting body morphologies except for Pl. aurantium which looks similar to P. mycophaga in fruiting morphology, but has amoebae that can make flagella in liquid medium. We built phylogenetic trees using nuclear small subunit ribosomal DNA sequences of 35 isolates from the genera Protostelium and Planoprotostelium and found that (1) the nonflagellated P. nocturnum and P. okumukumu branch basally in the genus Protostelium, (2) the flagellate, Pl. aurantium falls within the genus Protostelium in a monophyletic clade with the nominal variety, P. mycophaga var. crassipes, (3) the cultures initially identified as Protostelium mycophaga can be divided into at least three morphologically recognizable taxa, P. aurantium n. comb., P. apiculatum n. sp., and P. m. rodmani n. subsp., as well as a paraphyletic assemblage that includes the remainder of the P. mycophaga morphotype. These findings have implications for understanding the ecology, evolution, and diversity of these amoeboid organisms and for using these amoebae as models for other amoeboid groups.

Chitosan nanoparticles-loaded Citrus aurantium essential oil: a novel delivery system for preserving the postharvest quality of Agaricus bisporus.

BACKGROUND: One of the main problems in the button mushroom industry is the rapid deterioration of fruit bodies after harvest. Today, nanotechnology has become a more reliable technique to improve the quality of products in food packaging. In the present study, the effectiveness of chitosan nanoparticles containing Citrus aurantium essential oil on postharvest quality of white button mushroom was examined and compared to essential oil fumigation and control treatments. RESULTS: Based on high-resolution transmission electron microscopy and dynamic light scattering, nanoparticles exhibited a spherical shape of 20-60 nm diameter. The results revealed that the application of chitosan nanoparticles loaded with C. aurantium oil significantly decelerated the rate of color change, weight loss and firmness compared to fumigation with essential oil and control treatments. Treatment of fruit bodies with chitosan nanoparticles loaded with C. aurantium oil promoted the accumulation of phenolic compounds and ascorbic acid, and resulted in increases in catalase and superoxide dismutase and a decrease in polyphenol oxidase activities, as the highest antioxidant capacity was observed after 15 days of cold storage. CONCLUSION: This present research demonstrates that gradual release of C. aurantium essential oil from chitosan nanoparticles could be an effective and practical method for extending the shelf life of white button mushroom up to 15 days without significant decrease in antioxidant capacity. © 2018 Society of Chemical Industry.

Mushroom Emergence Detected by Combining Spore Trapping with Molecular Techniques.

Obtaining reliable and representative mushroom production data requires time-consuming sampling schemes. In this paper, we assessed a simple methodology to detect mushroom emergence by trapping the fungal spores of the fruiting body community in plots where mushroom production was determined weekly. We compared the performance of filter paper traps with that of funnel traps and combined these spore trapping methods with species-specific quantitative real-time PCR and Illumina MiSeq to determine the spore abundance. Significantly more MiSeq proportional reads were generated for both ectomycorrhizal and saprotrophic fungal species using filter traps than were obtained using funnel traps. The spores of 37 fungal species that produced fruiting bodies in the study plots were identified. Spore community composition changed considerably over time due to the emergence of ephemeral fruiting bodies and rapid spore deposition (lasting from 1 to 2 weeks), which occurred in the absence of rainfall events. For many species, the emergence of epigeous fruiting bodies was followed by a peak in the relative abundance of their airborne spores. There were significant positive relationships between fruiting body yields and spore abundance in time for five of seven fungal species. There was no relationship between fruiting body yields and their spore abundance at plot level, indicating that some of the spores captured in each plot were arriving from the surrounding areas. Differences in fungal detection capacity by spore trapping may indicate different dispersal ability between fungal species. Further research can help to identify the spore rain patterns for most common fungal species.IMPORTANCE Mushroom monitoring represents a serious challenge in economic and logistical terms because sampling approaches demand extensive field work at both the spatial and temporal scales. In addition, the identification of fungal taxa depends on the expertise of experienced fungal taxonomists. Similarly, the study of fungal dispersal has been constrained by technological limitations, especially because the morphological identification of spores is a challenging and time-consuming task. Here, we demonstrate that spores from ectomycorrhizal and saprotrophic fungal species can be identified using simple spore traps together with either MiSeq fungus-specific amplicon sequencing or species-specific quantitative real-time PCR. In addition, the proposed methodology can be used to characterize the airborne fungal community and to detect mushroom emergence in forest ecosystems.

The genus Retiboletus in China.

Species of the genus Retiboletus (Boletaceae, Boletales) in China are investigated based on morphology and phylogenetic analyses of DNA sequences from nuc rDNA internal transcribed spacer (ITS) and partial 28S regions and sequences from the translation elongation factor 1-a gene (tef1a). Six lineages are recovered among the collections studied. Five of these are documented and presented in the present paper, including three new species and two new combinations. The remaining species is not described due to the paucity of material. The specimens from China identified as "R. ornatipes" or "R. retipes" are in fact R. sinensis or R. kauffmanii, those labeled "R. griseus" are either R. fuscus or R. pseudogriseus A key to all known taxa of the genus is provided. Phylogenetic relationships of taxa within Retiboletus are partially resolved. A preliminary biogeographical analysis shows that allied species of Retiboletus between eastern Asia and North/Central America are common but there are no Retiboletus species common to both continents. Species of Retiboletus in Japan and southern China are conspecific or closely related.

Systematics of the ectomycorrhizal genus Lactarius in the Rocky Mountain alpine zone.

Lactarius (Russulales) is an important component of ectomycorrhizal fungal communities in cold-dominated contiguous arctic and disjunct alpine habitats where it associates primarily with Betula, Dryas and Salix However, little is known of this genus in the central and southern Rocky Mountain alpine zone (3000-3900 m) of North America. Molecular phylogenetic analyses of nuc rDNA ITS1-5.8S-ITS2 (ITS barcode) and the second largest subunit of the RNA polymerase II gene (RPB2) partial sequences in conjunction with detailed morphological examination confirm at least six species occurring above treeline. Most have intercontinental distributions in North America and Eurasia according to molecular comparison with type material and collections from Europe, Fennoscandia, Svalbard and Alaska. Rocky Mountain collections of L. lanceolatus (subgenus Russularia), along with the type from Alaska are paraphyletic with respect to L. aurantiacus and North American taxa L. luculentus and L. luculentus v. laetus Rocky Mountain collections of L. nanus, L. glyciosmus, L. repraesentaneus and L. salicis-reticulatae (subgenus Piperites) all form clades with European material from type localities and other arctic-alpine habitats. The arctic-alpine L. pseudouvidus/L. brunneoviolaceus group appears to be a complex containing additional taxa. North American material originally described as part of this group is well-separated phylogenetically and is described here as L. pallidomarginatus sp. nov. Lactarius lanceolatus, L. nanus and L. salicis-reticulatae appear largely restricted to arctic-alpine habitats with Salix Lactarius glyciosmus and L. repraesentaneus occur in arctic-alpine, subalpine and boreal habitats with Betula and also Picea and possibly Salix for the latter. Species distributions are hypothesized to be shaped by host ranges, glaciation and long distance dispersal.

The evolution of defense mechanisms correlate with the explosive diversification of autodigesting Coprinellus mushrooms (Agaricales, Fungi).

Bursts of diversification are known to have contributed significantly to the extant morphological and species diversity, but evidence for many of the theoretical predictions about adaptive radiations have remained contentious. Despite their tremendous diversity, patterns of evolutionary diversification and the contribution of explosive episodes in fungi are largely unknown. Here, using the genus Coprinellus (Psathyrellaceae, Agaricales) as a model, we report the first explosive fungal radiation and infer that the onset of the radiation correlates with a change from a multilayered to a much simpler defense structure on the fruiting bodies. We hypothesize that this change constitutes a key innovation, probably relaxing constraints on diversification imposed by nutritional investment into the development of protective tissues of fruiting bodies. Fossil calibration suggests that Coprinellus mushrooms radiated during the Miocene coinciding with global radiation of large grazing mammals following expansion of dry open grasslands. In addition to diversification rate-based methods, we test the hard polytomy hypothesis, by analyzing the resolvability of internal nodes of the backbone of the putative radiation using Reversible-Jump MCMC. We discuss potential applications and pitfalls of this approach as well as how biologically meaningful polytomies can be distinguished from alignment shortcomings. Our data provide insights into the nature of adaptive radiations in general by revealing a deceleration of morphological diversification through time. The dynamics of morphological diversification was approximated by obtaining the temporal distribution of state changes in discrete traits along the trees and comparing it with the tempo of lineage accumulation. We found that the number of state changes correlate with the number of lineages, even in parts of the tree with short internal branches, and peaks around the onset of the explosive radiation followed by a slowdown, most likely because of the decrease in available niches.

Understanding the evolutionary processes of fungal fruiting bodies: correlated evolution and divergence times in the Psathyrellaceae.

Fruiting body evolution is one of the central topics in fungal evolutionary biology. A number of hypotheses have been developed to explain the contemporary diversity of fruiting body forms, but their evaluation has been hampered by the lack of well-sampled data sets and suitable statistical methods. Phylogenetic evidence of the physiological changes that accompany switches in fruiting body type is lacking, and very little is known about the age of major events of fruiting body evolution. Based on a new multigene phylogeny, by using Bayesian methods, we demonstrate the existence of correlation between a number of morphological features and switches from nondeliquescent to deliquescent (autodigesting) fruiting bodies in the mushroom family Psathyrellaceae. Our results show that switches in the anatomy of two types of spacer cells (cystidia and pseudoparaphyses) and basidia (bimorphic or monomorphic) as well as the structure of the mushroom cap follow the evolution of deliquescent fruiting bodies, which suggests strong functional linkage between these traits. We performed Bayes factor-based tests, referred hereafter to as evolutionary pathway test (EPT), to decide which of the correlated characters were gained first during evolution. The EPTs strongly suggest that deliquescence was gained first, followed after short waiting times by the other morphological features. Bayesian relaxed molecular clock analyses suggest that the various events of switching between fruiting body types occurred independently at various ages during the history of the family. The utility of two mushroom fossils (Archaemarasmius and Protomycena), the only ones with unambiguous taxonomic positions, for the calibration of agaric trees were also examined. Based on our results, we suggest that the evolutionary benefit of deliquescence may be prevention against desiccation via accelerated ontogeny of the fruiting body. Hypotheses regarding the functional significance of the correlated evolution are presented and discussed. Further, we argue that the changes in fruiting body types in mushrooms in general can be attributed to independent events (e.g., dispersal and adaptation) and not to particular geologic ages.

Molecular phylogeny of Heteroplacidium, Placidium, and related catapyrenioid genera (Verrucariaceae, lichen-forming Ascomycota).

PREMISE OF THE STUDY: Verrucariaceae is a fascinating lineage of lichenized fungi for which generic and species delimitation is problematic due to the scarcity of discriminating morphological characters. Members of this family inhabit rocks, but they further colonize soils, barks, mosses, and other lichens. Our aim is to contribute to the DNA-based inference of the Verrucariaceae tree of life and to investigate characters that could be useful for proposing a more natural classification. We focused on catapyrenioid genera, which are often part of biological soil crusts, a cryptogam-dominated ecosystem contributing to soil formation and stabilization in arid environments. Understanding their evolution and taxonomy is essential to assess their roles in these fragile and important ecosystems. METHODS: A multigene phylogeny of Verrucariaceae including catapyrenioid genera is presented. We further examined the phylogenetic relationships among members of Heteroplacidium and Placidium. The evolution of selected characters was inferred using the latter phylogeny. KEY RESULTS: Anthracocarpon and Involucropyrenium were closely related to Endocarpon. Placidium comprised two monophyletic clades sister to Heteroplacidium. Inferred ancestral states of diagnostic characters revealed that the type of medulla and the pycnidia location were homoplasious within the Placidium clade. In contrast, the presence of rhizines was a synapomorphy for Clavascidium. CONCLUSIONS: Our results provide new information on the usefulness of characters for delineating groups in Verrucariaceae. Taxonomic changes are proposed to reflect more natural groupings: Heteroplacidium podolepis is transferred to Placidium, and Clavascidium is recognized as a different genus. Eight new combinations are proposed for Clavascidium.

Basidiomycetous yeasts from boletales fruiting bodies and their interactions with the mycoparasite Sepedonium chrysospermum and the host fungus Paxillus.

Interactions between mushrooms, yeasts, and parasitic fungi are probably common in nature, but are rarely described. Bolete fruiting bodies are associated with a broad spectrum of microorganisms including yeasts, and they are commonly infected with filamentous mycoparasites of the genus Sepedonium (teleomorph Hypomyces). We report the isolation of 17 yeast strains from Paxillus and Xerocomus, 16 of which were obtained from the surface tissue, the primary site of Sepedonium infection. Phylogenetic analyses with the D1/D2 region of the 28S ribosomal gene and the internal transcribed spacers placed the yeasts as Rhodotorula, Rhodosporidium, and Mastigobasidium from the Pucciniomycotina, Cryptococcus, Cystofilobasidium, Holtermanniella, and Trichosporon from the Agaricomycotina, and Kluyveromyces from the Saccharomycotina including the first isolation of Rhodotorula graminis from Europe. To investigate the influence of the yeast strains on the mycoparasite and the host fungus, in vitro assays were conducted with Sepedonium chrysospermum and Paxillus involutus. Both S. chrysospermum growth inhibitory and stimulating yeast strains were detected among the isolates. The number of S. chrysospermum inhibitory yeast strains increased and the number of S. chrysospermum stimulatory yeast strains decreased in the presence of P. involutus in co-cultures. Low nutrient levels in the culture medium also led to an increased number of S. chrysospermum inhibitory yeast strains and ten yeasts inhibited the mycoparasite in spatial separation by a crosswall. Six yeast strains inhibited P. involutus in dual culture, and the inhibitory P. involutus yeast interactions increased to nine in the presence of S. chrysospermum. Our results suggest that the bolete-associated yeasts influence the growth of the mycoparasitic fungus, which may affect the health of the fruiting bodies.

Two new species of Hydnum with ovoid basidiospores: H. ovoideisporum and H. vesterholtii.

Two new species of Hydnum, characterized by slender Hydnum rufescens-like basidiomes and ovoid to broadly ellipsoid basidiospores, are described from the Iberian Peninsula based on morphological and ITS molecular data. Hydnum ovoideisporum is distinguished by pilei with deep orange tones and strong preference for calcareous soil. It is widespread in the Iberian-Mediterranean area. Hydnum vesterholtii is characterized by its ocher to light ocher pileus, and nearly all the collections were made in the Pyrenees. Both ovoid-spored species are monophyletic well supported groups in the maximum parsimony and Bayesian ITS phylogenies, while the remainder of the samples assigned to H. rufescens s.l. and having globose basidiospores split into six well supported clades. The need to typify the name Hydnum rufescens is discussed, and a provisional key is given for the European taxa of Hydnum.

A new species of Melanospora on truffles from China.

The new species Melanospora subterranea is described from China, based on morphological and phylogenetic analyses. This is the first record of Melanospora species parasitizing Chinese black and white truffles (Tuber indicum and T. huidongense), and its host range indicates that the new species might be a disease threat to commercially exploited European truffles including Tuber melanosporum and Tuber magnatum. Apparent polyphyly within the Ceratostomataceae can be explained at least in part by inadvertent sequencing of the host fungus instead of the parasite.

Lactarius subg. Plinthogalus: the European taxa and American varieties of L. lignyotus re-evaluated.

The European species Lactarius subg. Plinthogalus were subjected to a molecular phylogenetic analysis based on ITS, LSU and rpb2 sequences. Morphological characters of the species are discussed in the light of the phylogenetic results. In addition to a broad sampling within Europe, some Asian and North American taxa also were included in the analysis. Eight European species are confirmed molecularly: L. lignyotus, L. acris, L. azonites, L. pterosporus, L. ruginosus, L. romagnesii, L. fuliginosus and L. picinus. Except the sibling species L. fuliginosus and L. picinus, all are morphologically distinct. Our results suggest that L. fuliginosus is associated exclusively with broadleaf trees and L. picinus with conifers, but this putative difference in host specificity needs to be investigated further. Lactarius subruginosus turns out to be a synonym of either L. pterosporus or L. ruginosus. The position of Lactarius terenopus remains to be clarified. The North American taxa that are closely related to the European L. lignyotus (L. fallax, L. lignyotus var. canadensis, var. nigroviolascens, var. marginatus) are not resolved. Intercontinental conspecificity was demonstrated between Europe and northern Asia but was not found between Europe and southern Asia or between Europe and North America. A taxonomic subdivision of L. subg. Plinthogalus based on the height of the spore ornamentation should be rejected.

Corneroboletus, a new genus to accommodate the southeastern Asian Boletus indecorus.

Corneroboletus was erected in the Boletaceae to accommodate Boletus indecorus originally described from southeastern Asia. The mucilaginous surface of the basidioma, the ixohyphoepithelium pileipellis and the irregularly warty to irregularly bacillate ornamentation of basidiospores distinguish this fungus from other described species in Boletaceae. Phylogenetic placement of this fungus was investigated further with molecular data including LSU rRNA and concatenated alignment of nrLSU, 5.8S rRNA and rpb2 genes, and an independent lineage among existing genera of Boletaceae was suggested by our phylogenetic results. Consequently a description, illustrations and a comparison of Corneroboletus with allied taxa are presented.

A new Cortinarius of section Calochroi (Basidiomycota, Agaricomycetes) from Mediterranean Quercus woodlands (Italy).

A new species of Cortinarius, C. flavoaurantians sp. nov., is described from Italian Quercus woods based on both morphological and ITS rDNA data. This taxon is characterized by a yellowish pileus and cortina, a white universal veil and a pileipellis that reacts yellow-orange with KOH. Illustrations of the key micromorphological features and fresh basidiomata in situ are provided. Closely related species are discussed.

Cantharellaceae of Guyana I: new species, combinations and distribution records of Craterellus and a synopsis of known taxa.

Members of the Cantharellaceae (Cantharellales, Basidiomycota) are common ectomycorrhizal associates of the leguminous genus Dicymbe in the Pakaraima Mountains of Guyana. Eight distinct species or morphospecies currently are recognized in Craterellus Pers. or Cantharellus Adans. ex Fr. from Guyanese Dicymbe-dominated forests. We evaluated the systematics of these taxa with phylogenetic analyses of DNA sequence data from the nuclear ribosomal regions of the internal transcribed spacer (ITS) and 28S large subunit (LSU). The results of these analyses along with careful assessment of morphology let us described two new species, Craterellus atratoides sp. nov. and Craterellus strigosus sp. nov., redescribe Craterellus atratus (Corner) Yomyart et al. based on new material from Guyana, and propose a new combination in Craterellus for Cantharellus pleurotoides T.W. Henkel, Aime & S.L. Mill. Macroscopic illustrations are provided for two additional cantharelloid morphospecies confirmed in Craterellus, as well as the regionally endemic Cantharellus guyanensis Mont. Macromorphological, micromorphological and habitat data are provided for C. atratoides, C. strigosus and C. atratus, and ITS and LSU sequence data are provided for each of the eight known Guyanese taxa.

Three new ascomycetes from freshwater in China.

Three new freshwater ascomycetes, Diaporthe aquatica sp. nov. (Diaporthaceae), Ophioceras aquaticus sp. nov. (Magnaporthaceae) and Togninia aquatica sp. nov. (Togniniaceae), are described and illustrated based on morphological and molecular data (ITS, 18S, 28S rDNA sequences). Diaporthe aquatica is characterized by globose to subglobose, black ascomata with long necks, broadly cylindrical to obclavate asci, and small, ellipsoidal to fusiform, one-septate, hyaline ascospores; it is unusual among Diaporthe species in the fact that it lacks a stroma and has freshwater habitat. Ophioceras aquaticus is characterized by globose ascomata with a long beak, cylindrical, eight-spored asci with J- subapical rings and 3-5-septate filiform ascospores with slightly acute ends. Togninia aquatica is characterized by globose ascomata with long necks, clavate and truncate asci clustered on distinct ascogenous hyphae, and small, reniform, hyaline ascospores. Differences among the new taxa and similar species are discussed.

First report of the ectomycorrhizal status of boletes on the Northern Yucatan Peninsula, Mexico determined using isotopic methods.

Despite their prominent role for tree growth, few studies have examined the occurrence of ectomycorrhizal fungi in lowland, seasonally dry tropical forests (SDTF). Although fruiting bodies of boletes have been observed in a dry tropical forest on the Northern Yucatan Peninsula, Mexico, their occurrence is rare and their mycorrhizal status is uncertain. To determine the trophic status (mycorrhizal vs. saprotrophic) of these boletes, fruiting bodies were collected and isotopically compared to known saprotrophic fungi, foliage, and soil from the same site. Mean δ(15)N and δ(13)C values differed significantly between boletes and saprotrophic fungi, with boletes 8.0‰ enriched and 2.5‰ depleted in (15)N and (13)C, respectively relative to saprotrophic fungi. Foliage was depleted in (13)C relative to both boletes and saprotrophic fungi. Foliar δ(15)N values, on the other hand, were similar to saprotrophic fungi, yet were considerably lower relative to bolete fruiting bodies. Results from this study provide the first isotopic evidence of ectomycorrhizal fungi in lowland SDTF and emphasize the need for further research to better understand the diversity and ecological importance of ectomycorrhizal fungi in these forested ecosystems.

An axenic culture system for fruiting body formation by an edible bolete phylogenetically related to culinary-medicinal penny bun mushroom, Boletus edulis Bull.:Fr. strains from China.

The ability of two freshly isolated Boletus stains to fruit under axenic conditions was tested using different solid and liquid nutrient media. One strain (YNCX04) produced numerous primordia from which fruiting bodies, 12 mm and 10 mm in length, with grey, convex pilei, and yellow-white, clavate stipes developed between 15 and 30 d after inoculation of fungal mycelium onto a solid medium consisting of mineral salts, thiamine, glucose, potato, an extract of Cunninghamia lanceolata root, and agar. The other strain (YNB200) produced numerous primordia but no sporophores. Strain YNCX04 lost the ability to form fruiting bodies in axenic culture 6 mo after initial isolation but retained the ability to form primordia for up to 18 mo. Based on internal transcribed spacer sequencing data, strains YNB200 and YNCX04 formed a sub-cluster together with four previously designated Boletus edulis strains from China. Phylogenetic analysis placed the Chinese strains closer to B. aestivalis than to European and North American strains of B. edulis, although a 29-bp fragment specific to all the B. aestivalis strains was absent from all the Chinese strains. Furthermore, partial 18S rDNA sequences from strains YNB200 and YNCX04 exhibited 98% similarity with an 18S rDNA sequence from B. edulis strain Be3. Further molecular studies are indicated to more accurately establish the taxonomic positions ofF3 and F4-3, as well as the Chinese strains designated as B. edulis.

Can wood-decaying urban macrofungi be identified by using fuzzy interference system? An example in Central European Ganoderma species.

Ganoderma is a cosmopolitan genus of wood-decaying basidiomycetous macrofungi that can rot the roots and/or lower trunk. Among the standing trees, their presence often indicates that a hazard assessment may be necessary. These bracket fungi are commonly known for the crust-like upper surfaces of their basidiocarps and formation of white rot. Six species occur in central European urban habitats. Several of them, such as Ganoderma adspersum, G. applanatum, G. resinaceum and G. pfeifferi, are most hazardous fungi causing extensive horizontal stem decay in urban trees. Therefore, their early identification is crucial for correct management of trees. In this paper, a fast technique is tested for the determination of phytopathologically important urban macrofungi using fuzzy interference system of Sugeno type based on 13 selected traits of 72 basidiocarps of six Ganoderma species and compared to the ITS sequence based determination. Basidiocarps features were processed for the following situations: At first, the FIS of Sugeno 2 type (without basidiospore sizes) was used and 57 Ganoderma basidiocarps (79.17%) were correctly determined. Determination success increased to 96.61% after selecting basidiocarps with critical values (15 basidiocarps). These undeterminable basidiocarps must be analyzed by molecular methods. In a case, that basidiospore sizes of some basidiocarps were known, a combination of Sugeno 1 (31 basidiocarps with known basidiospore size) and Sugeno 2 (41 basidiocarps with unknown basidiospore size) was used. 84.72% of Ganoderma basidiocarps were correctly identified. Determination success increased to 96.83% after selecting basidiocarps with critical values (11 basidiocarps).

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

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