Ursula Peintner

Towards a unified paradigm for sequence‐based identification of fungi

The nuclear ribosomal internal transcribed spacer (ITS) region is the formal fungal barcode and in most cases the marker of choice for the exploration of fungal diversity in environmental samples. Two problems are particularly acute in the pursuit of satisfactory taxonomic assignment of newly generated ITS sequences: (i) the lack of an inclusive, reliable public reference data set and (ii) the lack of means to refer to fungal species, for which no Latin name is available in a standardized stable way. Here, we report on progress in these regards through further development of the UNITE database (http://unite.ut.ee) for molecular identification of fungi. All fungal species represented by at least two ITS sequences in the international nucleotide sequence databases are now given a unique, stable name of the accession number type (e.g. Hymenoscyphus pseudoalbidus|GU586904|SH133781.05FU), and their taxonomic and ecological annotations were corrected as far as possible through a distributed, third‐party annotation effort. We introduce the term ‘species hypothesis’ (SH) for the taxa discovered in clustering on different similarity thresholds (97–99%). An automatically or manually designated sequence is chosen to represent each such SH. These reference sequences are released (http://unite.ut.ee/repository.php) for use by the scientific community in, for example, local sequence similarity searches and in the QIIME pipeline. The system and the data will be updated automatically as the number of public fungal ITS sequences grows. We invite everybody in the position to improve the annotation or metadata associated with their particular fungal lineages of expertise to do so through the new Web‐based sequence management system in UNITE.

Multiple origins of sequestrate fungi related to Cortinarius (Cortinariaceae).

The aim of the present study was to investigate the phylogeny and evolution of sequestrate fungi (with gastroid or partially exposed basidiomes) in relation to their gilled relatives from the Cortinariaceae (Basidiomycetes). Phylogenetic analyses of 151 ITS sequences from 77 gilled species and 37 sequestrate taxa were performed using maximum parsimony and maximum likelihood methods. Results show that sequestrate basidiome forms occur in all three major ectomycorrhizal lineages of Cortinariaceae: the clades Cortinarius, Hebeloma/Hymenogaster/Naucoria, and Descolea. However, these forms do not appear within the saprobic outgroup Gymnopilus, indicating multiple origins of sequestrate forms from ectomycorrhizal ancestors. Additionally, within the Cortinarius clade sequestrate forms have multiple origins: emergent Cortinarius spp., Thaxterogaster, Quadrispora, Protoglossum, and two Hymenogaster spp. (H. remyi, H. sublilacinus) share common ancestors with Cortinarius spp., but these sequestrate genera are not closely related to each other (with exception of Thaxterogaster and Quadrispora). Hymenogaster sensu stricto, Setchelliogaster, and Descomyces were placed in the two other major clades. Thus, sequestrate taxa evolved independently many times within brown-spored Agaricales. Furthermore, emergent, secotioid, and gastroid forms have evolved independently from each other, and so are not necessarily intermediate forms. After their establishment, these apparently morphologically stable taxa show a tendency to radiate.

Ectomycorrhizal communities associated with Populus tremula growing on a heavy metal contaminated site

European aspen is one of the most widely distributed trees in Central Europe and is a typical early colonizer of poor and disturbed soils. However, little is known about ectomycorrhizal (ECM) fungi in these ecosystems. We examined the ECM community of European aspen growing on a heavily contaminated site in southern Austria by analysing ECM roots, sorting them into morphotypes, subjecting them to DNA extraction, PCR, and DNA sequencing. ECM root symbionts were sampled two times in 2004. During this time, the below-ground community structure was relatively stable; we found no evidence of taxa adapted to summer or autumn conditions and only two species varied widely in occurrence between soil horizons. The ECM fungal community was diverse (54 species), rich in Basidiomycota (43 species), and dominated by Cenococcum geophilum and fungi with corticoid basidiomes (e.g. Thelephoraceae).

Toward a better understanding of the infrageneric relationships in Cortinarius (Agaricales, Basidiomycota)

Research on the molecular systematics of Cortinarius, a species-rich mushroom genus with nearly global distribution, is just beginning. The present study explores infrageneric relationships using rDNA ITS and LSU sequence data. One large dataset of 132 rDNA ITS sequences and one combined da-taset with 54 rDNA ITS and LSU sequences were generated. Hebeloma was used as outgroup. Bayesian analyses and maximum-likelihood (ML) analyses were carried out. Bayesian phylogenetic inference performed equally well or better than ML, especially in large datasets. The phylogenetic analysis of the combined dataset with species representing all currently recognized subgenera recovered seven well-supported clades (Bayesian posterior probabilities BPP > 90%). These major clades are: /Myxacium s.l., /subg. Cortinarius, the /phlegmacioid clade (including the subclades /Phlegmacium and /Delibuti), the /calochroid clade (/Calochroi, /Ochroleuci and /Allutus), the /telamonioid clade (/Telamonia, /Orellani, /Anomali), /Dermocybe s.l. and /Myxotelamonia. Our results show that Cortinarius consists of many lineages, but the relationships among these clades could not be elucidated. On one hand, the low divergence in rDNA sequences can be held responsible for this; on the other hand, taxon sampling is problematic in Cortinarius phylogeny. Because of the incredibly high diversity (~2000 Cortinarius species), our sampling included <5% of the known species. By choosing type species of subgenera and sections, our sampling is strongly biased toward Northern Hemisphere taxa. More extensive taxon sampling, especially of species from the Southern Hemisphere, is essential to resolve the phylogeny of this important genus of ectomycorrhizal fungi.

Biodiversity and concentration of airborne fungi in a hospital environment

The biodiversity and concentration of airborne fungi were monitored over a period of 6 months in a special-care unit of a hospital. Air sampling was performed in a corridor that was also accessible to visitors and in an adjacent bone-marrow transplantation (BMT) unit using an air sampler and two isolation media. Altogether, 98 fungal species could be identified, among them Aspergillus fumigatus and A. terreus as well as 48 other species reported as potential pathogens. The average contamination values of the corridor air ranged from 124 to 485 cfu m−3. Neither the degree of fungal air contamination nor the species composition inside the special care unit differed from those found in the corridor. By means of data obtained with a light-activated sensor, a possible influence of human activities on diurnal changes of fungal propagule concentration was shown.

Mycobionts of Salix herbacea on a glacier forefront in the Austrian Alps

Dwarf willows (e.g. Salix herbacea) are among the earliest ectomycorrhizal (EM) plants colonising primary successional sites such as glacier forefronts in the Tyrolean Alps. EM of S. herbacea were sampled at the Rotmoos glacier forefront (Ötz Valley, Austria) three times a year during the growing season and once a year during winter when plants were covered with snow in 2005 and 2006. EM were investigated using morphological methods and by sequencing the rDNA ITS region. The degree of EM mycorrhization was high throughout both years (93%). We distinguished 21 EM morphotypes and identified 19 fungal species. Cenococcum geophilum, Sebacina spp., Tomentella spp. and Cortinarius spp. dominated the mycobiont community of S. herbacea. The observed species richness in this about 150-year-old soil was at least 59% of the estimated species richness. Fungal communities differed significantly between consecutive years, and spatial heterogeneity was high. These differences made it difficult to detect seasonal impacts. Abundances of C. geophilum EM increased throughout the 2-year sampling period. Sebacina incrustans EM were very abundant in 2005, but nearly disappeared in 2006, whilst its fruitbodies were still frequent in the sampling area. This suggests that the mycorrhizae were displaced from the roots by an outcompeting species, whereas the mycelium was still present in the soil.

The iceman's fungi

Among the numerous items of equipment with the ‘Iceman’, who died more than 5000 years ago on an alpine glacier, were three fungal objects: two different shaped fruitbody pieces of the polypore Piptoporus betulinus, each mounted separately on a leather thong, and, found in his girdle bag, a relatively large quantity of tinder material prepared from the ‘true tinder bracket’ Fomes fomentarius. A full description of these items and a chronological report on their identification is given. The question about the possible use of the fungi is discussed on the basis of a comprehensive collection of ethnomycological and pharmacological literature data.

First records of ectomycorrhizal Cortinarius species (Agaricales, Basidiomycetes) from tropical India and their phylogenetic position based on rDNA ITS sequences

Three new Cortinarius species, Cortinarius conopileus, C. keralensis, and C. phlegmophorus spp. nov., are described from Kerala State in southern India. This is the first record of ectomycorrhizal Cortinarius spp. in the tropical part of India. In addition to distinct morphological characters, the comparative analysis of rDNA ITS sequences of the collections from India and morphologically similar species support the recognition of these taxa as new species. Phylogenetic analyses demonstrate that the three Indian Cortinarius spp. belong to both larger subclades of the genus Cortinarius, clade/cortinarius and clade/telamonia. As supported by morphological and molecular data, C. phlegmophorus belongs to Cortinarius subgen. Myxacium sect. Defibulati. Based on classical morphological characters, both C. keralensis and C. conopileus are representatives of subgen. Telamonia. However, C. conopileus belongs to clade/obtusi, which is a well-supported subclade of clade/cortinarius. Thus, in contrast to classical taxonomy, the clade/obtusi represents an independent evolutionary origin of telamonioid taxa. This result is also reflected by the distinct morphological characters of taxa of clade/obtusi, namely the lamellar trama with ellipsoid inflated hyphae and the presence of cystidia. In contrast, C. keralensis is a typical member of clade/telamonia. Within/telamonia, only relationships of closely related taxa are resolved due to the low genetic divergence found in ITS sequences. Based on morphological and molecular criteria, C. keralensis is a distinct taxon of sect. Saturnini.

European medicinal polypores – A modern view on traditional uses

ETHNOPHARMACOLOGICAL RELEVANCE In particular five polypore species, i.e. Laetiporus sulphureus, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus, and Laricifomes officinalis, have been widely used in central European folk medicines for the treatment of various diseases, e.g. dysmenorrhoea, haemorrhoids, bladder disorders, pyretic diseases, treatment of coughs, cancer, and rheumatism. Prehistoric artefacts going back to over 5000 years underline the long tradition of using polypores for various applications ranging from food or tinder material to medicinal-spiritual uses as witnessed by two polypore species found among items of Ötzi, the Iceman. The present paper reviews the traditional uses, phytochemistry, and biological activity of the five mentioned polypores. MATERIALS AND METHODS All available information on the selected polypore taxa used in traditional folk medicine was collected through evaluation of literature in libraries and searches in online databases using SciFinder and Web of Knowledge. RESULTS Mycochemical studies report the presence of many primary (e.g. polysaccharides) and secondary metabolites (e.g. triterpenes). Crude extracts and isolated compounds show a wide spectrum of biological properties, such as anti-inflammatory, cytotoxic, and antimicrobial activities. CONCLUSIONS The investigated polypores possess a longstanding ethnomycological tradition in Europe. Here, we compile biological results which highlight their therapeutic value. Moreover, this work provides a solid base for further investigations on a molecular level, both compound- and target-wise.

Phylogeny of Rozites, Cuphocybe and Rapacea inferred from ITS and LSU rDNA sequences

Phylogenetic relationships of Rozites, Cuphocybe, and Rapacea were assessed using molecular phylogenetic approaches. These three genera are placed in Cortinariaceae and have been regarded as closely related to Cortinarius. Rozites includes more than 20 species, which are characterized by having both a membranaceous partial veil in the form of a persistent annulus and a membranaceous universal veil. Cuphocye (4 species) lacks an annulus or cortina, but has pigmented veil fibrils or scales. The monotypic genus Rapacea accommodates a distinct taxon with pale, nearly smooth and thick-walled basidiospores. We analyzed 56 sequences of the internal transcribed spacer region (ITS1, ITS2, and the intervening 5.8S rRNA gene) for nine species of Rozites, three species of Cuphocybe, 28 species of Cortinarius, Rapacea mariae and Protoglossum luteum. Two species of Hebeloma were used as outgroup. Large subunit (LSU) rDNA sequences from selected taxa were also analyzed. The results clearly demonstrate that Rozites species are nested within the clade/Cortinarius, and that Rozites is polyphyletic, suggesting that membranaceous veils have evolved several times in the genus Cortinarius. Also Rapacea and Cuphocybe are nested within Cortinarius, making the latter genus paraphyletic. Based on phylogenetic studies, Rozites, Cuphocybe and Rapacea are artificial genera and do not reflect natural relationships.