Concordance of multigene genealogy along with morphological evidence unveils five novel species and two new records of boletoid mushrooms (fungi) from India.

Agaricales, Russulales and Boletales are dominant orders among the wild mushrooms in Basidiomycota. Boletaceae, one of the major functional elements in terrestrial ecosystem and mostly represented by ectomycorrhizal symbionts of trees in Indian Himalaya and adjoining hills, are extraordinarily diverse and represented by numerous genera and species which are unexplored or poorly known. Therefore, their hidden diversity is yet to be revealed. Extensive macrofungal exploration by the authors to different parts of Himalaya and surroundings, followed by through morphological studies and multigene molecular phylogeny lead to the discovery of five new species of wild mushrooms: Leccinellum bothii sp. nov., Phylloporus himalayanus sp. nov., Phylloporus smithii sp. nov., Porphyrellus uttarakhandae sp. nov., and Retiboletus pseudoater sp. nov. Present communication deals with morphological details coupled with illustrations and phylogenetic inferences. Besides, Leccinellum sinoaurantiacum and Xerocomus rugosellus are also reported for the first time from this country.

Responses of a soil fungal community to severe windstorm damages in an old silver fir stand.

Forests are increasingly threatened by climate change and the Anthropocene seems to have favored the emergence and adaptation of pathogens. Robust monitoring methods are required to prevent biodiversity and ecosystems losses, and this imposes the choice of bioindicators of habitat health. Fungal communities are increasingly recognized as fundamental components in nearly all natural and artificial environments, and their ecosystem services have a huge impact in maintaining and restoring the functionality of ecosystems. We coupled metabarcoding and soil analyses to infer the dynamics of a fungal community inhabiting the old silver fir stand in Vallombrosa (Italy), which is known to be afflicted by both Armillaria and Annosum root rot. The forest was affected in 2015, by a windstorm which caused a partial falling and uprooting of trees. The remaining stand, not affected by the windstorm, was used as a comparison to infer the consequences of the ecosystem disturbance. We demonstrated that the abundance of pathogens alone is not able to explain the soil fungal differences shown by the two areas. The fungal community as a whole was equally rich in the two areas, even if a reduction of the core ectomycorrhizal mycobiome was observed in the wind-damaged area, accompanied by the increase of wood saprotrophs and arbuscular mycorrhizas. We hypothesize a reshaping of the fungal community and a potentially ongoing re-generation of its functionalities. Our hypothesis is driven by the evidence that key symbiotic, endophytic, and saprotrophic guilds are still present and diversified in the wind-damaged area, and that dominance of single taxa or biodiversity loss was not observed from a mycological point of view. With the present study, we aim at providing evidence that fungal communities are fundamental for the monitoring and the conservation of threatened forest ecosystems.

Four Novel Species and Two New Records of Boletes from India.

Repeated macrofungal explorations, followed by thorough examination of species through morphology and molecular phylogeny, have made it clear that European and American names of wild mushrooms were inadvertently misapplied quite often to Asian lookalikes by mycologists/taxonomists in the past. Therefore, in order to reveal this mushroom treasure, in recent years, taxonomical research on wild mushrooms has been intensified in Asian countries, including India, by undertaking a combined approach of morpho-taxonomy and multigene molecular phylogeny. Boletoid mushrooms (Boletaceae) are no exception. While working on boletoid mushrooms of the Indian Himalayas, authors recently came across six interesting species of boletoid mushrooms. In the present communication, four novel species, namely Leccinellum binderi, Cyanoboletus paurianus, Xerocomus uttarakhandae, and Xerocomellus himalayanus, are established based on morphology and molecular phylogenetic estimations. Moreover, Cyanoboletus macroporus and Xerocomus fraternus are also reported here for the first time in India.

New Insights into Lichenization in Agaricomycetes Based on an Unusual New Basidiolichen Species of Omphalina s. str.

The genus Omphalina is an ideal genus for studying the evolutionary mechanism of lichenization. Based on molecular phylogeny using ITS and nuLSU sequences by means of Bayesian and maximum likelihood analyses and morphological examination, combining the existence of green algae in basidiomata stipe and a Botrydina-type vegetative thallus, we described a bryophilous new basidiolichen species, Omphalina licheniformis, from a residential area of Jiangxi Province, China. This finding of unusual new basidiolichen species updated our understanding of the delimitation of Omphalina, indicating that both non-lichen-forming and lichen-forming fungal species are included simultaneously. The presence of algal cells in the basidiomata should receive more attention, as this would be helpful to distinguish more potential basidiolichens and explore the cryptic species diversity. This work provides new insights and evidence for understanding the significance of lichenization during the evolution of Agaricomycetes.

Reappraisal of the Genus Exsudoporus (Boletaceae) Worldwide Based on Multi-Gene Phylogeny, Morphology and Biogeography, and Insights on Amoenoboletus.

The boletoid genera Butyriboletus and Exsudoporus have recently been suggested by some researchers to constitute a single genus, and Exsudoporus was merged into Butyriboletus as a later synonym. However, no convincing arguments have yet provided significant evidence for this congeneric placement. In this study, we analyze material from Exsudoporus species and closely related taxa to assess taxonomic and phylogenetic boundaries between these genera and to clarify species delimitation within Exsudoporus. Outcomes from a multilocus phylogenetic analysis (ITS, nrLSU, tef1-α and rpb2) clearly resolve Exsudoporus as a monophyletic, homogenous and independent genus that is sister to Butyriboletus. An accurate morphological description, comprehensive sampling, type studies, line drawings and a historical overview on the nomenclatural issues of the type species E. permagnificus are provided. Furthermore, this species is documented for the first time from Israel in association with Quercus calliprinos. The previously described North American species Exsudoporus frostii and E. floridanus are molecularly confirmed as representatives of Exsudoporus, and E. floridanus is epitypified. The eastern Asian species Leccinum rubrum is assigned here to Exsudoporus based on molecular evidence, and a new combination is proposed. Sequence data from the original material of the Japanese Boletus kermesinus were generated, and its conspecificity with L. rubrum is inferred as formerly presumed based on morphology. Four additional cryptic species from North and Central America previously misdetermined as either B. frostii or B. floridanus are phylogenetically placed but remain undescribed due to the paucity of available material. Boletus weberi (syn. B. pseudofrostii) and Xerocomus cf. mcrobbii cluster outside of Exsudoporus and are herein assigned to the recently described genus Amoenoboletus. Biogeographic distribution patterns are elucidated, and a dichotomous key to all known species of Exsudoporus worldwide is presented.

Systemic effects of Tuber melanosporum inoculation in two Corylus avellana genotypes.

Roots of the European hazelnut (Corylus avellana L.), i.e., one of the most economically important nut species, form symbiosis with ectomycorrhizal (ECM) fungi, including truffles. Although physical interactions only occur in roots, the presence of mycorrhizal fungi can lead to metabolic changes at a systemic level, i.e., in leaves. However, how root colonization by ECM fungi modifies these processes in the host plant has so far not been widely studied. This work aimed to investigate the response in two C. avellana genotypes, focusing on leaves from plants inoculated with the black truffle Tuber melanosporum Vittad. Transcriptomic profiles of leaves of colonized plants were compared with those of non-colonized plants, as well as sugar and polyphenolic content. Results suggested that T. melanosporum has the potential to support plants in stressed conditions, leading to the systemic regulation of several genes involved in signaling and defense responses. Although further confirmation is needed, our results open new perspectives for future research aimed to highlight novel aspects in ECM symbiosis.

Localized reshaping of the fungal community in response to a forest fungal pathogen reveals resilience of Mediterranean mycobiota.

Mediterranean forests are facing the impact of pests such as the soilborne Phytophthora cambivora, the causal agent of Ink disease, and this impact is made more severe by global changes. The status and resilience of the soil microbial ecosystem in areas with such a disturbance are little known; however, the assessment of the microbial community is fundamental to preserve the ecosystem functioning under emerging challenges. We profile soil fungal communities in a chestnut stand affected by ink disease in Italy using metabarcoding, and couple high-throughput sequencing with physico-chemical parameters and dendrometric measurements. Since the site also includes an area where the disease symptoms seem to be suppressed, we performed several analyses to search for determinants that may contribute to such difference. We demonstrate that neither pathogen presence nor trees decline associate with the reduction of the residing community diversity and functions, but rather with microbial network reshaping through substitutions and new interactions, despite a conservation of core taxa. We predict interactions between taxa and parameters such as soil pH and C/N ratio, and suggest that disease incidence may also relate with disappearance of pathogen antagonists, including ericoid- and ectomycorrhizal (ECM) fungi. By combining metabarcoding and field studies, we infer the resilient status of the fungal community towards a biotic stressor, and provide a benchmark for the study of other threatened ecosystems.

Taxonomic Re-Examination of Nine Rosellinia Types (Ascomycota, Xylariales) Stored in the Saccardo Mycological Collection.

In a recent monograph on the genus Rosellinia, type specimens worldwide were revised and re-classified using a morphological approach. Among them, some came from Pier Andrea Saccardo's fungarium stored in the Herbarium of the Padova Botanical Garden. In this work, we taxonomically re-examine via a morphological and molecular approach nine different Roselliniasensu Saccardo types. ITS1 and/or ITS2 sequences were successfully obtained applying Illumina MiSeq technology and phylogenetic analyses were carried out in order to elucidate their current taxonomic position. Only the ITS1 sequence was recovered for Rosellinia areolata, while for R. geophila, only the ITS2 sequence was recovered. We proposed here new combinations for Rosellinia chordicola, R. geophila and R. horridula, while for R. ambigua, R. areolata, R. australis, R. romana and R. somala, we did not suggest taxonomic changes compared to the current ones. The name Rosellinia subsimilis Sacc. is invalid, as it is a later homonym of R. subsimilis P. Karst. & Starbäck. Therefore, we introduced Coniochaeta dakotensis as a nomen novum for R. subsimilis Sacc. This is the first time that these types have been subjected to a molecular study. Our results demonstrate that old types are an important source of DNA sequence data for taxonomic re-examinations.

Squamanitaceae and three new species of Squamanita parasitic on Amanita basidiomes.

The systematic position of the enigmatically mycoparasitic genus Squamanita (Agaricales, Basidiomycota) together with Cystoderma, Phaeolepiota, Floccularia, and Leucopholiota is largely unknown. Recently they were recognized as Squamanitaceae, but previous studies used few DNA markers from a restricted sample of taxa from the family and lacked a formal taxonomic treatment. In this study, with newly generated sequences of the type of the genus Squamanita, S. schreieri, and several additional species of the family, the phylogeny is reinvestigated with a concatenated (18S-5.8S-nrLSU-RPB2-TEF1-α) dataset. This study reveals that Cystoderma, Phaeolepiota, Squamanita, Floccularia, and Leucopholiota are a monophyletic clade with strong statistical support in Bayesian analysis and form Squamanitaceae. Phaeolepiota nested within Cystoderma; Squamanita, Leucopholiota, and Floccularia clustered together as two monophyletic subclades; and Squamanita was present as a monophyletic clade with strong statistical support in both Maximum Likelihood and Bayesian analyses. The family name Squamanitaceae is formally emended and a detailed taxonomic treatment is presented to accommodate the five genera. Meanwhile, another concatenated (18S-ITS-nrLSU-RPB2-TEF1-α) dataset is used to investigate phylogenetic relationships and species delimitation in Squamanita. Our data indicates that "S. umbonata" from the Northern hemisphere forms two species complexes, one complex includes six specimens from North America, Europe, and East Asia, the other includes two specimens from Central America and North America respectively. Futhermore, species of Squamanita can parasitize species of Amanita, besides other fungal species. Squamanita mira parasitizes A. kitamagotake (A. sect. Caesareae), while S. orientalis and S. sororcula are parasites of species belonging to the A. sepiacea complex (A. sect. Validae). "Squamanita umbonata" from Italy occurs on A. excelsa (A. sect. Validae). Three new species of Squamanita from East Asia, viz. S. mira, S. orientalis and S. sororcula are documented with morphological, multi-gene phylogenetic, and ecological data, along with line drawings and photographs, and compared with similar species. A key for identification of the global Squamanita species is provided.

Testing spore amyloidity in Agaricales under light microscope: the case study of Tricholoma.

Although species of the genus Tricholoma are currently considered to produce inamyloid spores, a novel standardized method to test sporal amyloidity (which involves heating the sample in Melzer's reagent) showed evidence that in the tested species of this genus, which belong in all 10 sections currently recognized from Europe, the spores are amyloid. In two species, T. josserandii and T. terreum, the spores are also partly dextrinoid. This result provides strong indication that a positive reaction of the spores in Melzer's reagent could be a character shared by all genera in Tricholomataceae s. str.

First Report of a Neotropical Agaric (Lepiota spiculata, Agaricales, Basidiomycota) Containing Lethal α-Amanitin at Toxicologically Relevant Levels.

A recent collection of Lepiota spiculata from the Dominican Republic is presented here. Macro- and micromorphological features of L. spiculata are described in detail, and its evolutionary (phylogenetic) position within Lepiota sect. Ovisporae, in the subincarnata/brunneoincarnata clade, is assessed on the basis of a combined nrLSU + nrITS + rpb2 + tef1 analysis. Additionally, high levels of deadly amatoxins were detected and quantified in L. spiculata for the first time by HPLC analysis; in particular, α-amanitin was found at concentrations up to approximately 4 mg/g dry weight, which render L. spiculata a potentially lethal mushroom, if ingested.

Outstanding Pinkish Brown-Spored Neotropical Boletes: Austroboletus subflavidus and Fistulinella gloeocarpa (Boletaceae, Boletales) from the Dominican Republic.

The occurrence of Austroboletus subflavidus and Fistulinella gloeocarpa is documented from the Dominican Republic. The latter species is reported for the first time outside its original locality in Martinique, extending the geographic range for this uncommon pinkish-spored bolete. A detailed morphological description is provided for each species and accompanied by color pictures of fresh basidiomes in habitat and line drawings of the main anatomical features. Both species represent independent lineages within their respective genera based on phylogenetic inference. In addition, A. subflavidus clusters in a sister lineage to the core Austroboletus clade (Austroboletus clade I) here named as Austroboletus clade II. In order to confirm the accuracy of species identification, their identity and relationships were subjected to multilocus phylogenetic analyses of three gene markers (ITS, nrLSU, RPB2) including genetic material already available in public databases. Austroboletus subflavidus is a widely distributed species in North and Central America, whereas F. gloeocarpa is apparently highly localized and seems to appear sparingly in the Dominican Republic, Martinque, and southern Florida. Comparisons with morphologically similar and molecularly inferred allied species are also presented and discussed.

Neoboletusantillanus sp. nov. (Boletaceae), first report of a red-pored bolete from the Dominican Republic and insights on the genus Neoboletus.

Neoboletusantillanus sp. nov. appears to be the only red-pored bolete known from the Dominican Republic to date. It is reported as a novel species to science based on collections gathered in a neotropical lowland mixed broadleaved woodland. A detailed morphological description, color images of fresh basidiomes in habitat and line drawings of the main anatomical features are provided and relationships with phylogenetically and phenotypically similar taxa are discussed. Three genomic regions (nrITS, nrLSU/28S and rpb2) have been sequenced in order to reinforce the recognition of the new species and to elucidate its taxonomic affiliation within Neoboletus.

Looking for Lepiotapsalion Huijser & Vellinga (Agaricales, Agaricaceae).

Lepiotapsalion is fully described based on a recent collection from Sardinia (Italy) and the holotype. NrITS- and nrLSU-based phylogeny demonstrates that sequences deposited in GenBank as "L.psalion" and generated from two Dutch and one Chinese collections are not conspecific with the holotype and represent two distinct, undescribed species. These species are here proposed as Lepiotarecondita sp. nov. and Lepiotasinorecondita ad int.

A new species of Rhodocybe sect. Rufobrunnea (Entolomataceae, Agaricales) from Italy.

Rhodocybe fumanellii is described from Italy as a new species based both on morphological and molecular nrITS/nrLSU data. It belongs in sect. Rufobrunnea and is characterised by massive tricholomatoid basidiomata with reddish-brown tinges, adnate and crowded lamellae, an enlarged stipe base with long rhizomorphs, long sinuose slender cheilocystidia, ellipsoid basidiospores and the presence of caulocystidia. Drawings of the main micromorphological features as well as a colour photograph of fresh basidiomata in situ are provided and its morphological relationships with allied species are discussed.

Diversity of polypores in the Dominican Republic: Pseudowrightoporia dominicana sp. nov. (Hericiaceae, Russulales).

The new species Pseudowrightoporia dominicana is described from the Dominican Republic based on morphological and molecular data (nrITS and nrLSU sequence analyses). It is mainly characterised by pileate basidiomata with a bright pinkish context and a di-trimitic hyphal system. Phylogenetically, it is sister to the African species P. gillesii and to the Asiatic P. japonica.

Phylogenetic studies in Genabea, Myrmecocystis, and related genera.

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.

Two new species and one new record of the genus Tylopilus (Boletaceae) from Indian Himalaya with morphological details and phylogenetic estimations.

Tylopilus himalayanus and T. pseudoballoui are described as new species from two Himalayan states (Sikkim and Uttarakhand) in India. Tylopilus himalayanus is characterised by a unique combination of features: reddish- or brownish-grey to purplish-grey then brown to reddish-brown or darker pileus, absence of olive or violet tinges on stipe surface, angular pores, stipe without reticulum or rarely with a faint reticulum restricted to the very apex, bitter taste of the context and positive macrochemical colour reaction of the stipe context with KOH (dark orange) and FeSO4 (dark green), medium sized (10.9-14.4 × 3.9-4.9 µm) basidiospores and occurrence under coniferous trees; T. pseudoballoui is distinguished by orange-yellow to brown-yellow sticky pileus, pale yellow pore surface with pinkish hues that turns pale to greyish-orange on bruising; angular pores, stipe concolorous to pileus with pruinose but never reticulate surface, ixocutis pattern of pileipellis and occurrence under broadleaf trees. Another species, T. neofelleus, which was reported earlier from China and Japan, was also collected from Sikkim and reported for the first time from India. All three species are described with morphological details and two-locus based (nrLSU and nrITS) phylogenetic data.

Gliophorusglutinosus sp. nov. (Hygrophoraceae, Agaricales) from Eastern Himalayan region of India.

An interesting species of the genus Gliophorus (sect.Glutinosae), collected from Sikkim Himalaya in India, is described here as G.glutinosus sp. nov. after thorough morphological examination and phylogenetic analysis. The species is mainly characterised by its strongly glutinous basidiomata throughout, particularly on the twisted stipe, decurrent lamellae with glutinous edge, gelatinised cheilocystidia, presence of pleuropseudocystidia and absence of clamps in hyphae of the pileipellis. This communication includes detailed morphological description, illustrations, comparison with the allied taxa, nrITS based phylogeny of this novel taxon and a key to the species under Gliophorussect.Glutinosae.