Alien and cryptogenic fungi and oomycetes in Austria: an annotated checklist (2nd edition).

Fungal invasions can have far-reaching consequences, and despite increasing relevance, fungi are notoriously underrepresented in invasion science. Here, we present the second annotated checklist for alien and cryptogenic fungi and oomycetes in Austria. This list contains 375 taxa of which 278 are classified as established; compared to the first checklist from 2002, this amounts to an almost five-fold increase and the number of decade-wise first records is steadily rising since the mid-twentieth century. The introduction pathway is unclear for the vast majority of taxa, while the main means of spread within the country is unassisted secondary spread. Fungi were predominantly introduced from the Northern Hemisphere, especially North America and Temperate Asia. Rates of newly recorded alien fungi differ among phyla; the majority belongs to the Ascomycota, which experienced an 9.6-fold increase in numbers. Orders found most frequently are powdery mildews (Erysiphales, Ascomycota), downy mildews (Peronosporales, Oomycota), agarics (Agaricales, Basidiomycota), Mycosphaerellales (Ascomycota), rusts (Pucciniales, Basidiomycota) and Pleosporales (Ascomycota). The majority (about 80%) of the taxa are plant pathogens, while animal pathogens are few but severely affecting their native hosts. The dominance of pathogens in our checklist underlines the need of better tackling fungal invasions-especially in the light of emerging infectious diseases-and highlights potential knowledge gaps for ectomycorrhizal and saprobic alien fungi, whose invasion processes are often much more inconspicuous. Our results show that fungal invasions are a phenomenon of increasing importance, and collaborative efforts are needed for advancing the knowledge and management of this important group. Supplementary Information: The online version contains supplementary material available at 10.1007/s10530-022-02896-2.

New cytochalasans from an endophytic Xylaria species associated with Costa Rican Palicourea elata (Rubiaceae).

Four new leucine-derived cytochalasans, possessing a 5,6,5,8-ring (1) and a 5,6,11-ring core (2-4), were isolated from a cultivated endophytic fungus Xylaria sp. strain WH2D4 (Xylariaceae). This fungus was isolated from leaves of the neotropical tree species Palicourea elata (Sw.) Borhidi (Rubiaceae) collected in Costa Rica. The chemical structures were determined by employing IR, MS as well as 1D- and 2D-NMR experiments. The stereochemistry at C-15 of compound 4 was determined by quantum calculations. The isolated compounds did not affect germination and growth of Trichoderma reesei and the opportunistic human fungal pathogen T. longibrachiatum.

Note on the genus Nemania (Xylariaceae) - first records and a new species of the genus from Iran.

In a survey of xylarialean fungi in northern Iran, some specimens attributable to the genus Nemania were collected, cultured and sequenced. Morphological evidence and phylogenetic analyses of a combined ITS, LSU, RPB2 and TUB2 gene dataset confirmed the presence of Nemaniadiffusa and N.serpens in Iran for the first time. Furthermore, the new species N.hyrcana, which shows similarities to N.subaenea and its putative synonym N.plumbea, but significantly differs from the latter in its DNA sequences, was encountered. All species are illustrated, described and discussed. In the phylogenetic analyses, for the first time, the overlooked ex-type ITS sequences of the neotype of the generic type, N.serpens and that of the holotype of N.prava, were added to a multi-gene matrix of Nemania. This revealed that the two accessions of N.serpens (HAST 235 and CBS 679.86), for which multigene data are available in GenBank, are misidentified, while the Iranian accession of N.serpens has an almost identical ITS sequence to the neotype, confirming its morphological species identification. The two previously accepted species of Euepixylon, E.udum and E.sphaeriostomum, are embedded within Nemania and are revealed as close relatives of N.serpens, supporting the inclusion of Euepixylon in Nemania.

Resolution of the Hypoxylon fuscum Complex (Hypoxylaceae, Xylariales) and Discovery and Biological Characterization of Two of Its Prominent Secondary Metabolites.

Hypoxylon, a large, cosmopolitan genus of Ascomycota is in the focus of our current poly-thetic taxonomic studies, and served as an excellent source for bioactive secondary metabolites at the same time. The present work concerns a survey of the Hypoxylon fuscum species complex based on specimens from Iran and Europe by morphological studies and high performance liquid chromatography coupled to mass spectrometry and diode array detection (HPLC-MS-DAD). Apart from known chemotaxonomic markers like binaphthalene tetrol (BNT) and daldinin F, two unprece-dented molecules were detected and subsequently isolated to purity by semi preparative HPLC. Their structures were established by nuclear-magnetic resonance (NMR) spectroscopy as 3'-malonyl-daldinin F (6) and pseudofuscochalasin A (4). The new daldinin derivative 6 showed weak cytotoxicity towards mammalian cells but bactericidal activity. The new cytochalasin 4 was compared to cytochalasin C in an actin disruption assay using fluorescence microscopy of human osteo-sarcoma U2OS cells, revealing comparable activity towards F-actin but being irreversible compared to cytochalasin C. Concurrently, a multilocus molecular phylogeny based on ribosomal and proteinogenic nucleotide sequences of Hypoxylon species resulted in a well-supported clade for H. fuscum and its allies. From a comparison of morphological, chemotaxonomic and phylogenetic evidence, we introduce the new species H. eurasiaticum and H. pseudofuscum.

Discovery of a new species of the Hypoxylon rubiginosum complex from Iran and antagonistic activities of Hypoxylon spp. against the Ash Dieback pathogen, Hymenoscyphus fraxineus, in dual culture.

During a survey of xylarialean fungi in Northern Iran, several specimens that showed affinities to the Hypoxylon rubiginosum complex were collected and cultured. A comparison of their morphological characters, combined with a chemotaxonomic study based on high performance liquid chromatography, coupled with diode array detection and mass spectrometry (HPLC-DAD/MS) and a multi-locus phylogeny based on ITS, LSU, rbp2 and tub2 DNA sequences, revealed a new species here described as Hypoxylon guilanense. In addition, Hypoxylon rubiginosum sensu stricto was also encountered. Concurrently, an endophytic isolate of the latter species showed strong antagonistic activities against the Ash Dieback pathogen, Hymenoscyphus fraxineus, in a dual culture assay in our laboratory. Therefore, we decided to test the new Iranian fungi for antagonistic activities against the pathogen, along with several cultures of other Hypoxylon species that are related to H. rubiginosum. Our results suggest that the antagonistic effects of Hypoxylon spp. against Hym. fraxineus are widespread and that they are due to the production of antifungal phomopsidin derivatives in the presence of the pathogen.

Altitudinal upwards shifts in fungal fruiting in the Alps.

Many plant and animal species are changing their latitudinal and/or altitudinal distributions in response to climate change, but whether fungi show similar changes is largely unknown. Here, we use historical fungal fruit body records from the European Alps to assess altitudinal changes in fungal fruiting between 1960 and 2010. We observe that many fungal species are fruiting at significantly higher elevations in 2010 compared to 1960, and especially so among soil-dwelling fungi. Wood-decay fungi, being dependent on the presence of one or a few host trees, show a slower response. Species growing at higher elevations changed their altitudinal fruiting patterns significantly more than lowland species. Environmental changes in high altitudes may lead to proportionally stronger responses, since high-altitude species live closer to their physiological limit. These aboveground changes in fruiting patterns probably mirror corresponding shifts in belowground fungal communities, suggesting parallel shifts in important ecosystem functions.

European mushroom assemblages are darker in cold climates.

Thermal melanism theory states that dark-colored ectotherm organisms are at an advantage at low temperature due to increased warming. This theory is generally supported for ectotherm animals, however, the function of colors in the fungal kingdom is largely unknown. Here, we test whether the color lightness of mushroom assemblages is related to climate using a dataset of 3.2 million observations of 3,054 species across Europe. Consistent with the thermal melanism theory, mushroom assemblages are significantly darker in areas with cold climates. We further show differences in color phenotype between fungal lifestyles and a lifestyle differentiated response to seasonality. These results indicate a more complex ecological role of mushroom colors and suggest functions beyond thermal adaption. Because fungi play a crucial role in terrestrial carbon and nutrient cycles, understanding the links between the thermal environment, functional coloration and species' geographical distributions will be critical in predicting ecosystem responses to global warming.

Open-source data reveal how collections-based fungal diversity is sensitive to global change.

PREMISE OF THE STUDY: Fungal diversity (richness) trends at large scales are in urgent need of investigation, especially through novel situations that combine long-term observational with environmental and remotely sensed open-source data. METHODS: We modeled fungal richness, with collections-based records of saprotrophic (decaying) and ectomycorrhizal (plant mutualistic) fungi, using an array of environmental variables across geographical gradients from northern to central Europe. Temporal differences in covariables granted insight into the impacts of the shorter- versus longer-term environment on fungal richness. RESULTS: Fungal richness varied significantly across different land-use types, with highest richness in forests and lowest in urban areas. Latitudinal trends supported a unimodal pattern in diversity across Europe. Temperature, both annual mean and range, was positively correlated with richness, indicating the importance of seasonality in increasing richness amounts. Precipitation seasonality notably affected saprotrophic fungal diversity (a unimodal relationship), as did daily precipitation of the collection day (negatively correlated). Ectomycorrhizal fungal richness differed from that of saprotrophs by being positively associated with tree species richness. DISCUSSION: Our results demonstrate that fungal richness is strongly correlated with land use and climate conditions, especially concerning seasonality, and that ongoing global change processes will affect fungal richness patterns at large scales.

Fungal Systematics and Evolution: FUSE 5.

Thirteen new species are formally described: Cortinarius brunneocarpus from Pakistan, C. lilacinoarmillatus from India, Curvularia khuzestanica on Atriplex lentiformis from Iran, Gloeocantharellus neoechinosporus from China, Laboulbenia bernaliana on species of Apenes, Apristus, and Philophuga (Coleoptera, Carabidae) from Nicaragua and Panama, L. oioveliicola on Oiovelia machadoi (Hemiptera, Veliidae) from Brazil, L. termiticola on Macrotermes subhyalinus (Blattodea, Termitidae) from the DR Congo, Pluteus cutefractus from Slovenia, Rhizoglomus variabile from Peru, Russula phloginea from China, Stagonosporopsis flacciduvarum on Vitis vinifera from Italy, Strobilomyces huangshanensis from China, Uromyces klotzschianus on Rumex dentatus subsp. klotzschianus from Pakistan. The following new records are reported: Alternaria calendulae on Calendula officinalis from India; A. tenuissima on apple and quince fruits from Iran; Candelariella oleaginescens from Turkey; Didymella americana and D. calidophila on Vitis vinifera from Italy; Lasiodiplodia theobromae causing tip blight of Dianella tasmanica 'variegata' from India; Marasmiellus subpruinosus from Madeira, Portugal, new for Macaronesia and Africa; Mycena albidolilacea, M. tenuispinosa, and M. xantholeuca from Russia; Neonectria neomacrospora on Madhuca longifolia from India; Nothophoma quercina on Vitis vinifera from Italy; Plagiosphaera immersa on Urtica dioica from Austria; Rinodina sicula from Turkey; Sphaerosporium lignatile from Wisconsin, USA; and Verrucaria murina from Turkey. Multi-locus analysis of ITS, LSU, rpb1, tef1 sequences revealed that P. immersa, commonly classified within Gnomoniaceae (Diaporthales) or as Sordariomycetes incertae sedis, belongs to Magnaporthaceae (Magnaporthales). Analysis of a six-locus Ascomycota-wide dataset including SSU and LSU sequences of S. lignatile revealed that this species, currently in Ascomycota incertae sedis, belongs to Pyronemataceae (Pezizomycetes, Pezizales).

Explaining European fungal fruiting phenology with climate variability.

Here we assess the impact of geographically dependent (latitude, longitude, and altitude) changes in bioclimatic (temperature, precipitation, and primary productivity) variability on fungal fruiting phenology across Europe. Two main nutritional guilds of fungi, saprotrophic and ectomycorrhizal, were further separated into spring and autumn fruiters. We used a path analysis to investigate how biogeographic patterns in fungal fruiting phenology coincided with seasonal changes in climate and primary production. Across central to northern Europe, mean fruiting varied by approximately 25 d, primarily with latitude. Altitude affected fruiting by up to 30 d, with spring delays and autumnal accelerations. Fruiting was as much explained by the effects of bioclimatic variability as by their large-scale spatial patterns. Temperature drove fruiting of autumnal ectomycorrhizal and saprotrophic groups as well as spring saprotrophic groups, while primary production and precipitation were major drivers for spring-fruiting ectomycorrhizal fungi. Species-specific phenology predictors were not stable, instead deviating from the overall mean. There is significant likelihood that further climatic change, especially in temperature, will impact fungal phenology patterns at large spatial scales. The ecological implications are diverse, potentially affecting food webs (asynchrony), nutrient cycling and the timing of nutrient availability in ecosystems.

Fungal Systematics and Evolution: FUSE 3.

The present study introduces seven new species, one new combination, one new variety and several interesting taxonomical notes and/or geographical records. Most of the new taxa are Ascomycetes, but the study also includes a new variety of a Basidiomycete. Novel species include Gyromitra khanspurensis (Discinaceae, Pezizales, Pezizomycetes) from Pakistan growing near Cedrus deoadara and Paramyrothecium guiyangense and Paramyrothecium verruridum (Stachybotriaceae, Hypocreales, Sordariomycetes) both isolated from soil in China. New species from South Africa are Sclerostagonospora elegiae on culm litter of Elegia equisetacea, Sclerostagonospora fusiformis on culm litter of Thamnochortus spicigerus, Sclerostagonospora pinguis on culm litter of Cannomois virgata and Sclerostagonospora sulcata on culm litter of Ischyrolepis subverticellata (Phaeosphaeriaceae, Pleosporales, Dothideomycetes). Hapalocystis berkeleyi var. kickxii with its basionym Hypoxylon kickxii is shown to be a taxon on species level and thus recombined as Hapalocystis kickxii (Sydowiellaceae, Diaporthales, Sordariomycetes), and it is lecto- and epitypified. The new variety Pluteus romellii var. luteoalbus (Pluteaceae, Agaricales, Agaricomycetes) growing on a mossy fallen stem of a deciduous tree is described from Czech Republic. Cortinarius scaurocaninus (Cortinariaceae, Agaricales, Agaricomycetes) is new for Austria, Humicola grisea (Chaetomiaceae, Sordariales, Sordariomycetes) is an interesting new record for Chile. Two taxa are reported as new for Turkey: the lichenicolous fungus Opegrapha parasitica (Opegraphaceae, Arthoniales, Arthoniomycetes) growing partly immersed in the thallus of Aspicilia and the lichen Rinodina zwackhiana (Physciaceae, Teloschistales, Lecanoromycetes) from calcareous rock. Finally, Xerula strigosa (Physalacriaceae, Agaricales, Agaricomycetes), described from China, is confirmed to be present also in Pakistan.

Rapid genetic detection of ingested Amanita phalloides.

Mushrooms are often poorly digested by humans. Thus, their remains (tissues, spores) may persist in the gastrointestinal tract and can be detected in feces several days after mushroom consumption. In this report, we present protocols for the rapid PCR-based detection of fungal traces in a variety of complex samples. Novel primers were designed to amplify portions of ribosomal DNA from deadly poisonous European members of the genus Amanita, namely the death cap (A. phalloides), the destroying angel (A. virosa) and the fool's mushroom (A. verna), respectively. Assay sensitivity was sufficient to discover diluted DNA traces in amounts below the genomic content of a single target mushroom cell. Specificity testing was performed with DNA extracts from a variety of mushroom species. Template amplification was exclusively observed with intended targets and it was not compromised by a vast excess of non-target DNA (i.e. DNA from human and human fecal origin, respectively). A series of experiments was conducted with prepared specimens in order to follow the course of mushroom food processing and digestion. Amplification by direct PCR was successful with raw, fried and digested mixed mushrooms. To improve assay performance with fecal samples, a rapid protocol for sample pre-processing (including water-ether sedimentation and bead beating) and a modified PCR reaction mix were applied. Thereby, it was possible to detect the presence of A. phalloides DNA in spiked feces as well as in clinical samples (vomit, stool) from two independent cases of suspected mushroom poisoning.

Iteratively refined guide trees help improving alignment and phylogenetic inference in the mushroom family Bolbitiaceae.

Reconciling traditional classifications, morphology, and the phylogenetic relationships of brown-spored agaric mushrooms has proven difficult in many groups, due to extensive convergence in morphological features. Here, we address the monophyly of the Bolbitiaceae, a family with over 700 described species and examine the higher-level relationships within the family using a newly constructed multilocus dataset (ITS, nrLSU rDNA and EF1-alpha). We tested whether the fast-evolving Internal Transcribed Spacer (ITS) sequences can be accurately aligned across the family, by comparing the outcome of two iterative alignment refining approaches (an automated and a manual) and various indel-treatment strategies. We used PRANK to align sequences in both cases. Our results suggest that--although PRANK successfully evades overmatching of gapped sites, referred previously to as alignment overmatching--it infers an unrealistically high number of indel events with natively generated guide-trees. This 'alignment undermatching' could be avoided by using more rigorous (e.g. ML) guide trees. The trees inferred in this study support the monophyly of the core Bolbitiaceae, with the exclusion of Panaeolus, Agrocybe, and some of the genera formerly placed in the family. Bolbitius and Conocybe were found monophyletic, however, Pholiotina and Galerella require redefinition. The phylogeny revealed that stipe coverage type is a poor predictor of phylogenetic relationships, indicating the need for a revision of the intrageneric relationships within Conocybe.

Warming-induced shift in European mushroom fruiting phenology.

In terrestrial ecosystems, fungi are the major agents of decomposition processes and nutrient cycling and of plant nutrient uptake. Hence, they have a vital impact on ecosystem processes and the terrestrial carbon cycle. Changes in productivity and phenology of fungal fruit bodies can give clues to changes in fungal activity, but understanding these changes in relation to a changing climate is a pending challenge among ecologists. Here we report on phenological changes in fungal fruiting in Europe over the past four decades. Analyses of 746,297 dated and geo-referenced mushroom records of 486 autumnal fruiting species from Austria, Norway, Switzerland, and the United Kingdom revealed a widening of the annual fruiting season in all countries during the period 1970-2007. The mean annual day of fruiting has become later in all countries. However, the interspecific variation in phenological responses was high. Most species moved toward a later ending of their annual fruiting period, a trend that was particularly strong in the United Kingdom, which may reflect regional variation in climate change and its effects. Fruiting of both saprotrophic and mycorrhizal fungi now continues later in the year, but mycorrhizal fungi generally have a more compressed season than saprotrophs. This difference is probably due to the fruiting of mycorrhizal fungi partly depending on cues from the host plant. Extension of the European fungal fruiting season parallels an extended vegetation season in Europe. Changes in fruiting phenology imply changes in mycelia activity, with implications for ecosystem function.

Molecular studies on terricolous microfungi reveal novel anamorphs of two Tuber species.

This study reports novel terricolous mitosporic fungal morphs nested in the genus Tuber according to molecular phylogenetic analysis. Fungal DNA was amplified directly from field-collected anamorph samples. Nuclear ribosomal DNA (nrDNA) sequences including the ITS regions and the D1 and D2 domains of the LSU identify the anamorphs as mitosporic Tuber borchii and Tuber oligospermum. The link of the novel anamorphs to the genus Tuber is confirmed by the comparative analysis of five collections from four sampling sites. Ectomycorrhizas with characteristic features of Tuber borchii ectomycorrhizas were found in the soil volume collected with one of the mitosporic T. borchii collections. A nrDNA sequence amplified from these ectomycorrhizae is identical with the corresponding anamorph sequence. The possible role of the newly discovered anamorphs in the Tuber life-cycle and the potential significance of anamorphs for the propagation of ectomycorrhizal fungi are discussed.

Assemblage structure, species richness, abundance, and distribution of fungal fruit bodies in a seven year plot-based survey near Vienna.

Almost 900 species were encountered during a seven year survey of fungal fruit bodies in 13 forests and grasslands plots measuring about 1 ha. The data were comparable to those from a Swiss forest plot monitored for 21 years. Species richness and abundance were almost linearly correlated on a log/log scale. Abundance and yearly frequency of species were tightly correlated. The geographic range of species may also be related to these two parameters. Rare species scored low on abundance, yearly frequency and geographic range. About half of the species were rare, and occurred in only one out of seven years. The two major functional groups of saprotrophic and mycorrhizal species behaved similarly over the years, as in the Swiss study. Annual variation was high for species richness alone and for richness combined with abundances. The log transformed species richness of plots correlated well with parameters that take the abundances or yearly frequencies of species into account. If other mushroom assemblages show similar regularities, it may be possible to omit abundance counts in future surveys. The species composition of the plots varied strongly over the years. Many species did not reach their maximum abundance in the richest year. Species showed their maximum abundances in different plots in the same year more often than expected by chance. Thus the presence and abundance of species depended on factor(s) other than the general productivity of a year, and years showed (a) different aspect(s) than productivity alone. Long-term surveys are important for understanding the structure of mushroom assemblages and their biodiversity.