Phylogenetic overview of the genus Genea (Pezizales, Ascomycota) with an emphasis on European taxa.

We constructed a comprehensive phylogeny of the genus Genea, with new molecular data from samples collected in several countries in temperate and Mediterranean Europe, as well as North America. Type specimens and authentic material of most species were examined to support identifications. The molecular identity of the most common species in Genea was compared with nuc rDNA internal transcribed spacer (ITS), D1-D2 domains of 28S nuc rDNA (28S rDNA) and translation elongation factor 1-α ene (TEF1) profiles of 10 recently proposed taxa, G. brunneocarpa, G. compressa, G. dentata, G. fageticola, G. lobulata, G. oxygala, G. pinicola, G. pseudobalsleyi, G. pseudoverrucosa and G. tuberculata, supporting their status as distinct species. Genea mexicana and G. thaxteri on the one hand and G. sphaerica and G. lespiaultii on the other are closely related. Multiple lineages were recorded for G. verrucosa and G. fragrans, but we found no morphological traits to discriminate among them, so we tentatively interpreted them as cryptic species. A key to species of the genus Genea is provided to facilitate identification. We provide macroscopic images of fresh specimens and of representative spores of most species. Finally, we conducted a molecular analysis of the divergence time for Genea and discuss the implications of our results.

Sequestrate fungi from Patagonian Nothofagus forests: Cystangium (Russulaceae, Basidiomycota).

Six species of Cystangium, a genus of sequestrate taxa related to Russula, were collected in Patagonia (Argentina and Chile) during autumn 2001. Two species, C. depauperatum Singer & A.H. Sm. and C. nothofagi (E. Horak) Trappe, Castellano & T. Lebel, were already known from this region, while four new species, C. domingueziae, C. gamundiae, C. grandihyphatum and C. longisterigmatum, are described, illustrated and a key to the species is provided. In addition, sequences of the ITS (rDNA) region were obtained to explore the phylogenetic relationships of our South American Cystangium species.

Morchella australiana sp. nov., an apparent Australian endemic from New South Wales and Victoria.

An abundant fruiting of a black morel was encountered in temperate northwestern New South Wales (NSW), Australia, during a mycological survey in Sep 2010. The site was west of the Great Dividing Range in a young, dry sclerophyll forest dominated by Eucalyptus and Callitris north of Coonabarabran in an area known as the Pilliga Scrub. Although the Pilliga Scrub is characterized by frequent and often large, intense wildfires, the site showed no sign of recent fire, which suggests this species is not a postfire morel. Caps of the Morchella elata-like morel were brown with blackish ridges supported by a pubescent stipe that became brown at maturity. Because no morel has been described as native to Australia, the collections were subjected to multilocus molecular phylogenetic and morphological analyses to assess its identity. Results of these analyses indicated that our collection, together with collections from NSW and Victoria, represented a novel, genealogically exclusive lineage, which is described and illustrated here as Morchella australiana T. F. Elliott, Bougher, O'Donnell & Trappe, sp. nov.

Rare but widespread: A systematic revision of the truffle-forming genera Destuntzia and Kjeldsenia and the formation of a new genus, Hosakaea.

Here we present the results of taxonomic and systematic study of the rare truffle-forming genera Destuntzia and Kjeldsenia. Truffle-forming fungi are difficult to study due to their reduced morphological features and their cryptic, hypogeous fruiting habits. The rare occurrence of Destuntzia and Kjeldsenia further compounds these difficulties due to the lack of adequate material for study. Recent forays in North Carolina and Tennessee recovered new specimens of another rarely collected fungus, Octaviania purpurea. Morphological and phylogenetic analysis revealed that Octaviania purpurea is a member of the genus Destuntzia, and this led us to reevaluate the taxonomic status and systematic relationships of other Destuntzia species. We performed a multilocus phylogenetic analysis of Destuntzia specimens deposited in public fungaria, including all available type material, and environmental sequences from animal scat and soil. Our analyses indicate that Destuntzia is a member of the family Claustulaceae within the order Phallales and is a close relative of Kjeldsenia. Results of our phylogenetic analysis infer that three species originally described in the genus Destuntzia are members of the genus Kjeldsenia. We propose three new combinations in Kjeldsenia to accommodate these species as well as a new combination in Destuntzia to accommodate Octaviania purpurea. We also describe a new genus in Claustulaceae, Hosakaea, to accommodate a closely affiliated species, Octaviania violascens. Finally, we transfer the genus Destunzia into the family Claustulaceae and emend the description of the family. The newly proposed combinations in Destuntzia and Kjeldsenia significantly expand the known geographic ranges of both genera. The data from metabarcode analysis of scat and soil also reveal several additional undescribed species that expand these ranges well beyond those suggested by basidiomata collections. Systematic placement of Destuntzia in the saprotrophic order Phallales suggests that this genus is not ectomycorrhizal, and the ecological implications of this systematic revision are discussed.

New North American truffles (Tuber spp.) and their ectomycorrhizal associations.

Recent surveys of belowground fungal biodiversity in México and USA have revealed many undescribed truffle species, including many in the genus Tuber. Here we describe seven new species: Tuber beyerlei, T. castilloi, T. guevarai, T. lauryi, T. mexiusanum, T. miquihuanense and T. walkeri. Phylogenetic analyses place these species within the Maculatum group, an understudied clade of small truffles with little apparent economic value. These species are among the more taxonomically challenge-ing in the genus. We collected Tuber castilloi, T. mexiusanum and T. guevarai as fruit bodies and ectomycorrhizae on Quercus spp. in forests of eastern México. Tuber mexiusanum has a particularly broad geographic range, being collected in eastern USA under Populus deltoides and in Minnesota and Iowa in mixed hardwood forests. T. walkeri is described from the upper midwestern USA, and T. lauryi and T. beyerlei occur in the western USA.

Australasian sequestrate fungi 18: Solioccasus polychromus gen. & sp. nov., a richly colored, tropical to subtropical, hypogeous fungus.

Solioccasus polychromus gen. & sp. nov., the most brightly colored hypogeous fungus known, is described from Papua New Guinea and tropical northern Australia south into subtropical forests along the Queensland coast and coastal mountains to near Brisbane. Phylogenetic analysis of molecular data places it as a sister genus to Bothia in the Boletineae, a clade of predominantly ectomycorrhizal boletes. Ectomycorrhizal trees, such as members of the Myrtaceae (Eucalyptus, Corymbia, Lophostemon, Melaleuca spp.) and Allocasuarina littoralis, were present usually in mixture or in some cases dominant, so we infer some or all of them to be among the ectomycorrhizal hosts of S. polychromus.

The enigmatic truffle Fevansia aurantiaca is an ectomycorrhizal member of the Albatrellus lineage.

Fevansia aurantiaca is an orange-colored truffle that has been collected infrequently in the Pacific Northwest of the USA. This sequestrate, hypogeous fungus was originally thought to be related to the genera Rhizopogon or Alpova in the Boletales, but the large, inflated cells in the trama and the very pale spore mass easily segregated it from these genera. To date, no molecular phylogenetic studies have determined its closest relatives. F. aurantiaca was originally discovered in leaf litter beneath Pinaceae, leading Trappe and Castellano (Mycotaxon 75:153-179, 2000) to suggest that it is an ectomycorrhizal symbiont of various members of the Pinaceae. However, without direct ecological or phylogenetic data, it is impossible to confirm the trophic mode of this truffle species. In this study, we combined phylogenetic analysis of the ITS and 28S ribosomal DNA with data on microscopic morphology to determine that F. aurantiaca is a member of the Albatrellus ectomycorrhizal lineage (Albatrellaceae, Russulales).

Cryptolepiota, a new sequestrate genus in the Agaricaceae with evidence for adaptive radiation in western North America.

Phylogenetic analyses based on nLSU and ITS sequence data indicate that the sequestrate genus Gigasperma is polyphyletic. Gigasperma cryptica, which is known only from New Zealand, has affinities with the Cortinariaceae whereas G. americanum and two additional undescribed taxa from western North America are derived from Lepiota within the Agaricaceae. The three North American taxa appear to be recently evolved and are closely related. They occur in similar environments and form a well supported clade indicating that adaptive radiation has occurred within this group of fungi. An independent genus with sequestrate fructifications, Cryptolepiota is proposed to accommodate the three species in this clade. Cryptolepiota microspora and C. mengei are described as new, and G. americanum is transferred to Cryptolepiota. Gigasperma cryptica is illustrated and compared with the species of Cryptolepiota.

The dawn of symbiosis between plants and fungi.

The colonization of land by plants relied on fundamental biological innovations, among which was symbiosis with fungi to enhance nutrient uptake. Here we present evidence that several species representing the earliest groups of land plants are symbiotic with fungi of the Mucoromycotina. This finding brings up the possibility that terrestrialization was facilitated by these fungi rather than, as conventionally proposed, by members of the Glomeromycota. Since the 1970s it has been assumed, largely from the observation that vascular plant fossils of the early Devonian (400 Ma) show arbuscule-like structures, that fungi of the Glomeromycota were the earliest to form mycorrhizas, and evolutionary trees have, until now, placed Glomeromycota as the oldest known lineage of endomycorrhizal fungi. Our observation that Endogone-like fungi are widely associated with the earliest branching land plants, and give way to glomeromycotan fungi in later lineages, raises the new hypothesis that members of the Mucoromycotina rather than the Glomeromycota enabled the establishment and growth of early land colonists.

Terfezia disappears from the American truffle mycota as two new genera and Mattirolomyces species emerge.

Reexamination and molecular phylogenetic analyses of American Terfezia species and Mattirolomyces tiffanyae revealed that their generic assignments were wrong. Therefore we here propose these combinations: Mattirolomyces spinosus comb. nov. (≡ Terfezia spinosa), Stouffera longii gen. & comb. nov. (≡ Terfezia longii) and Temperantia tiffanyae gen. & comb. nov. (≡ Mattirolomyces tiffanyae). In addition we describe a new species, Mattirolomyces mexicanus spec. nov. All species belong to the Pezizaceae. Based on these results Terfezia is not known from North America, Mattirolomyces is represented by two species and two new monotypic genera are present.

Greetings from belowground: two new species of truffles in the genus Pachyphlodes (Pezizaceae, Pezizales) from México.

Pachyphlodes is a lineage of ectomycorrhizal, hypogeous, sequestrate ascomycete fungi native to temperate and subtropical forests in the Northern Hemisphere. Pachyphlodes species form ectomycorrhizae mainly with Fagales hosts. Here we describe two new species of Pachyphlodes, P.brunnea, and P.coalescens, based on morphological and phylogenetic analysis. Pachyphlodesbrunnea is distributed in the states of Tamaulipas and Nuevo León in northern México, occurring with Quercus and Juglans species. It is characterized by its dark brown peridium, white gleba, and spores with capitate columns. Pachyphlodescoalescens is distributed in the states of Michoacán and Tlaxcala in central and southwestern México co-occurring with Quercus and is distinguished by its reddish-brown peridium, light yellow gleba, and spore ornamentation. Both species, along with P.marronina, constitute the Marronina clade. This clade contains North American species characterized by a brown peridium and spores ornamented with capitate spines to coalesced spine tips that form a partial perispore.

A global meta-analysis of Tuber ITS rDNA sequences: species diversity, host associations and long-distance dispersal.

Truffles (Tuber) are ectomycorrhizal fungi characterized by hypogeous fruitbodies. Their biodiversity, host associations and geographical distributions are not well documented. ITS rDNA sequences of Tuber are commonly recovered from molecular surveys of fungal communities, but most remain insufficiently identified making it difficult to determine whether these sequences represent conspecific or novel taxa. In this meta-analysis, over 2000 insufficiently identified Tuber sequences from 76 independent studies were analysed within a phylogenetic framework. Species ranges, host associates, geographical distributions and intra- and interspecific ITS variability were assessed. Over 99% of the insufficiently identified Tuber sequences grouped within clades composed of species with little culinary value (Maculatum, Puberulum and Rufum). Sixty-four novel phylotypes were distinguished including 36 known only from ectomycorrhizae or soil. Most species of Tuber showed 1-3% intraspecific ITS variability and >4% interspecific ITS sequence variation. We found 123 distinct phylotypes based on 96% ITS sequence similarity and estimated that Tuber contains a minimum of 180 species. Based on this meta-analysis, species in Excavatum, Maculatum and Rufum clades exhibit preference for angiosperm hosts, whereas those in the Gibbosum clade are preferential towards gymnosperms. Sixteen Tuber species (>13% of the known diversity) have putatively been introduced to continents or islands outside their native range.

Kalapuya brunnea gen. & sp. nov. and its relationship to the other sequestrate genera in Morchellaceae.

Kalapuya is described as a new, monotypic truffle genus in the Morchellaceae known only from the Pacific northwestern United States. Its relationship to other hypogeous genera within Morchellaceae is explored by phylogenetic analysis of the ribosomal LSU and EF1alpha protein coding region. The type species, K. brunnea, occurs in Douglas-fir forests up to about 50 y old on the west slope of the Cascade Range in Oregon and in the Coastal Ranges of Oregon and northern California. It has a roughened, warty, reddish brown to brown peridium, a solid whitish gleba that develops grayish brown mottling as the spores mature, and produces a cheesy-garlicky odor at maturity. Its smooth, ellipsoid spores resemble those of Morchella spp. but are much larger. The four hypogeous genera known in the Morchellaceae, Kalapuya, Fischerula, Imaia and Leucangium, are distinct from the epigeous genera Morchella and Verpa, but it is uncertain whether they resulted from a single transition to a hypogeous fruiting habit or from multiple independent transitions. Kalapuya, locally known as the Oregon brown truffle, has been commercially harvested for culinary use.

Improved resolution of major clades within Tuber and taxonomy of species within the Tuber gibbosum complex.

Tuber gibbosum Harkn., described from northern California, originally was thought to be a single, variable species that fruited from autumn through winter to spring. It has become popular as a culinary truffle in northwestern USA, where it is commercially harvested. Morphological studies suggested it might be a complex that includes at least two species. We conducted morphological and phylogenetic studies of the complex to determine how many species it might contain and how they differed morphologically, geographically and seasonally. We also provide the first LSU phylogeny for the genus Tuber. Phylogenetic analyses resolve nine major clades in the genus with high bootstrap support and distinguish the Gibbosum clade from the Aestivum, Excavatum, Macrosporum, Magnatum, Melanosporum, Puberulum, Rufum and Spinoreticulatum clades. Further analyses of ITS and LSU regions revealed four distinct species in the Gibbosum complex. Although morphologically similar the four species differ in spore size and shape and in peridial anatomy. These species share the synapomorphy of having suprapellis hyphae with distinctive, irregular wall swellings at maturity; we have not seen this hyphal type in any other Tuber spp. worldwide. The three new species are named and described as T. bellisporum Bonito & Trappe, T. castellanoi Bonito & Trappe and T. oregonense Trappe, Bonito & Rawlinson.

Suillus quiescens, a new species commonly found in the spore bank in California and Oregon.

Suillus quiescens sp. nov. is common under Pinus muricata on Santa Cruz and Santa Rosa Islands in the northern Channel Islands of California, and we subsequently found it fruiting at Point Reyes National Seashore on the central coast of California. Sequences from the internal transcribed spacer region show that it is distinct from all 44 species of Suillus tested, and features of its morphology separate it from all other unsequenced species. Suillus quiescens has a broader distribution than coastal California because it also was encountered as ectomycorrhizae on roots of pine seedlings from the eastern Sierra Nevada, coastal Oregon and the southern Cascade Mountains. The reason it had not been identified from these areas might be due to its resemblance to S. brevipes at maturity or it might be a rare fruiter that persists in the spore bank.

The new truffle genus Babosia and a new species of Stouffera from semiarid grasslands of Hungary.

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.

Diversity and habitat relationships of hypogeous fungi. III. Factors influencing the occurrence of fire-adapted species.

Among the huge array of hypogeous ectomycorrhizal fungi so far documented from Australia, six genera and more than 30 species occur within the family Mesophelliaceae, all of which show various adaptations for surviving in fire-prone landscapes. These mostly endemic fungi are critical to postfire reestablishment of regenerating vegetation, and their fruit-bodies provide essential food resources for diverse ground-dwelling fauna. We developed habitat models for five common representatives of the Mesophelliaceae based on repeat collections of their fruit-bodies from 136 study plots situated along a series of environmental gradients across the south-eastern mainland of Australia. At a meso- or landscape scale, temperature influenced the occurrence of Castoreum radicatum, Mesophellia clelandii and Nothocastoreum cretaceum, with the type of response varying. Below a threshold, C. radicatum preferred sites with cooler mean annual temperatures. In contrast, M. clelandii and N. cretaceum had optimal ranges of temperature, above and below which the probability of detecting them dropped. Also at a landscape scale, C. radicatum was more likely to be detected at sites with lower levels of precipitation during the driest quarter of the year. At a micro-site scale, M. clelandii and N. cretaceum were more likely to occur in stands with an intermediate number of host eucalypt stems, likely relating to successional age of the stand. Sites with a higher number of large fallen trees were more likely to have N. cretaceum, while sites with intermediate litter depths were more likely to have C. radicatum and M. clelandii. Mesophellia glauca and M. trabalis showed no consistent patterns. They are apparently the most broadly adaptable in terms of the independent variables tested. Although fire has been previously suggested to be heavily implicated in the life cycle of several members of the Mesophelliaceae, we found no relationship between time since disturbance by fire and other factors and likelihood of occurrence. Instead, other habitat attributes appeared to be more important in explaining their distribution. The complex and differing responses of the species of Mesophelliaceae studied here, to features of their environment, reinforce the need to manage multiple-use forest landscapes across the region for a diversity of attributes.

Cyclooxygenase-2 inhibitory and antioxidant compounds from the truffle Elaphomyces granulatus.

The ethanol extract of fruiting bodies of Elaphomyces granulatus, a truffle-like fungus, was evaluated for cyclooxygenase-2 (COX-2) enzyme inhibitory and antioxidant activities. Inhibition of COX-2 activity was evaluated in mouse macrophages (RAW 264.7). The extract of E. granulatus caused a 68% inhibition of COX-2 activity at 50 microg/mL. Bioassay-guided investigation led to the isolation and identification of two active compounds, syringaldehyde and syringic acid. Syringaldehyde moderately inhibited COX-2 activity with an IC(50) of 3.5 microg/mL, while syringic acid strongly inhibited COX-2 activity with an IC(50) of 0.4 microg/mL. The antioxidant activity of the extract and isolated compounds was evaluated in HL-60 cells by the DCFH-DA method. The extract of E. granulatus showed a potent antioxidant effect, with an IC(50) of 41 microg/mL. Of the pure compounds, syringic acid displayed a strong antioxidant activity, with an IC(50) of 0.7 microg/mL, while syringaldehyde showed no activity in the assay.

Lanostane-type triterpenes from the mushroom Astraeus pteridis with antituberculosis activity.

Bioassay-guided fractionation of an EtOH extract of the truffle-mimiking mushroom Astraeus pteridis led to the isolation and identification of three new (3-5) and two known (1, 2) lanostane triterpenes and phenylalanine betaine (6). The structures of the isolates were elucidated on the basis of 1D and 2D NMR spectroscopic data, HRESIMS results, and X-ray crystallographic analysis. Compounds 5 and 1 showed moderate activity against Mycobacterium tuberculosis with MIC values of 34.0 and 58.0 microg/mL, respectively.

Imaia, a new truffle genus to accommodate Terfezia gigantea.

Originally described from Japan by Sanshi Imai in 1933, the hypogeous ascomycete Terfezia gigantea was subsequently discovered in the Appalachian Mountains of the USA. Morphological, electron microscopic, and phylogenetic studies of specimens collected in both regions revealed that, despite this huge geographic disjunction, (1) the Japanese and Appalachian specimens are remarkably similar both in morphology and the sampled rDNA sequences, (2) the species unambiguously falls into the Morchellaceae and is separated from the genus Terfezia in the Pezizaceae, (3) its spores are much larger than those of Terfezia spp. and are enclosed in a unique, electron-semitransparent, amorphous epispore that appears to be permeated with minute, meandering strands or canals. In addition to the molecular phylogenetic results, the numerous nuclei in ascospores, the dome shaped, striate ascus septal plugs and the long cylindric Woronin bodies also strengthen the family assignment to the Morchellaceae. Moreover, the species occurs in moist, temperate forests as opposed to the xeric to arid habitats of other Terfezia spp. We propose the new, monotypic genus Imaia to accommodate the species.