Chance or Necessity-The Fungi Co-Occurring with Formica polyctena Ants.

Studies on carton nesting ants and domatia-dwelling ants have shown that ant-fungi interactions may be much more common and widespread than previously thought. Until now, studies focused predominantly on parasitic and mutualistic fungi-ant interactions occurring mostly in the tropics, neglecting less-obvious interactions involving the fungi common in ants' surroundings in temperate climates. In our study, we characterized the mycobiota of the surroundings of Formica polyctena ants by identifying nearly 600 fungal colonies that were isolated externally from the bodies of F. polyctena workers. The ants were collected from mounds found in northern and central Poland. Isolated fungi were assigned to 20 genera via molecular identification (ITS rDNA barcoding). Among these, Penicillium strains were the most frequent, belonging to eight different taxonomic sections. Other common and widespread members of Eurotiales, such as Aspergillus spp., were isolated very rarely. In our study, we managed to characterize the genera of fungi commonly present on F. polyctena workers. Our results suggest that Penicillium, Trichoderma, Mucor, Schwanniomyces and Entomortierella are commonly present in F. polyctena surroundings. Additionally, the high diversity and high frequency of Penicillium colonies isolated from ants in this study suggest that representatives of this genus may be adapted to survive in ant nests environment better than the other fungal groups, or that they are preferentially sustained by the insects in nests.

New Endohyphal Relationships between Mucoromycota and Burkholderiaceae Representatives.

Mucoromycota representatives are known to harbor two types of endohyphal bacteria (EHB)-Burkholderia-related endobacteria (BRE) and Mycoplasma-related endobacteria (MRE). While both BRE and MRE occur in fungi representing all subphyla of Mucoromycota, their distribution is not well studied. Therefore, it is difficult to resolve the evolutionary history of these associations in favor of one of the following two alternative hypotheses explaining their origin: "early invasion" and "late invasion." Our main goal was to fill this knowledge gap by surveying Mucoromycota fungi for the presence of EHB. We screened 196 fungal strains from 16 genera using a PCR-based approach to detect bacterial 16S rRNA genes, complemented with fluorescence in situ hybridization (FISH) imaging to confirm the presence of bacteria within the hyphae. We detected Burkholderiaceae in ca. 20% of fungal strains. Some of these bacteria clustered phylogenetically with previously described BRE clades, whereas others grouped with free-living Paraburkholderia Importantly, the latter were detected in Umbelopsidales, which previously were not known to harbor endobacteria. Our results suggest that this group of EHB is recruited from the environment, supporting the late invasion scenario. This pattern complements the early invasion scenario apparent in the BRE clade of EHB.IMPORTANCE Bacteria living within fungal hyphae present an example of one of the most intimate relationships between fungi and bacteria. Even though there are several well-described examples of such partnerships, their prevalence within the fungal kingdom remains unknown. Our study focused on early divergent terrestrial fungi in the phylum Mucoromycota. We found that ca. 20% of the strains tested harbored bacteria from the family Burkholderiaceae Not only did we confirm the presence of bacteria from previously described endosymbiont clades, we also identified a new group of endohyphal Burkholderiaceae representing the genus Paraburkholderia We established that more than half of the screened Umbelopsis strains were positive for bacteria from this new group. We also determined that, while previously described BRE codiverged with their fungal hosts, Paraburkholderia symbionts did not.

Identification of Entomopathogenic Fungi as Naturally Occurring Enemies of the Invasive Oak Lace Bug, Corythucha arcuata (Say) (Hemiptera: Tingidae).

The oak lace bug (OLB), Corythucha arcuata (Hemiptera: Tingidae), was first identified as an invasive pest in Europe in northern Italy in 2000 and since then it has spread rapidly, attacking large forested areas in European countries. The OLB is a cell sap-sucking insect that is native to North America, with Quercus spp. as its main host. Its rapid expansion, successful establishment in invaded countries, and observations of more damage to hosts compared to native areas are most likely due to a lack of natural enemies, pathogens and competitors. In its native area, various natural enemies of OLBs have been identified; however, little is known about the occurrence and impact of OLB pathogens. None of the pathogenic fungi found on OLBs in natural conditions have been identified until now. In this study, we provide evidence of four entomopathogenic fungi that are naturally occurring on invasive OLBs found in infested pedunculate oak forests in eastern Croatia. On the basis of their morphology and multilocus molecular phylogeny, the fungi were identified as Beauveria pseudobassiana, Lecanicillium pissodis, Akanthomyces attenuatus and Samsoniella alboaurantium. The sequences generated for this study are available from GenBank under the accession numbers MT004817-MT004820, MT004833-MT004835, MT027501-MT27510, and MT001936-MT0011943. These pathogenic species could facilitate biological control strategies against OLBs.

The XVIII Congress of European Mycologists: Conference Report.

The 18th Congress of European Mycologists took place from 16 to 21 September 2019 in Warsaw and Bialowieza, Poland (Figure 1) [...].

New fossils of ascomycetous anamorphic fungi from Baltic amber.

Three new fossils of saprotrophic fungi are presented and described from Baltic amber, dated to Eocene epoch (Paleogene, upper to mid-Eocene). All belong to Ascomycota and are represented by hyphae as well as asexual reproduction structures allowing to assign them to present genera, respectively Periconia, Penicillium and Scopulariopsis. These material provide both the first and the oldest known fossil record of the mentioned taxa, making these data valuable for the knowledge about the evolutionary history of the Ascomycota.

Aureoboletus projectellus (Fungi, Boletales) - Occurrence data, environmental layers and habitat suitability models for North America and Europe.

Aureoboletus projectellus is a bolete species native to eastern North America, which has been introduced to Central Europe. Here we present summarized data about occurrence of the fungus in both disjunctive ranges based on (1) de novo georeferencing of herbarium specimens and occurrence reports; (2) information from peer-reviewed articles, mycological forums and websites; (3) personal observations and (4) from queries sent to Forest Districts and National Parks in Poland. Corresponding background data were acquired from public databases and include range of genus Pinus - obligatory mycorrhizal partner of A. projectellus - and WorldClim bioclimatic data. Both datasets were fit for purpose of range modelling, i.e. were represented as spatially compatible equal-area raster grids encompassing temperate forest biom in eastern North America and Europe. Additionally, maps of habitat suitability, reflecting association between occurrence and background data, were obtained using maximum entropy approach implemented in MaxEnt.

Psychronectria hyperantarctica, gen. nov., comb. nov., epitypification and phylogenetic position of an Antarctic bryophilous ascomycete.

The holotype of Thyronectria hyperantarctica was re-examined, redescribed, and compared with new collections of an Antarctic bryophilous ascomycete from a similar area. Because the condition of type material was insufficient for molecular studies, the authors designated an epitype from newly collected material with a high degree of morphological similarity to the holotype and paratype material. Phylogenetic analysis of the epitype revealed that its closest phylogenetic affinity was with the family Tilachlidiaceae and it formed a monophyletic group in this lineage within other collections of the species. Therefore, the new monotypic genus Psychronectria is described to accommodate Thyronectria hyperantarctica. The fungus is superficially similar to Thyronectria species in Nectriaceae, but the ascospores differ in color, size, and type of septation.

Metschnikowia cf. typographi and other pathogens from the bark beetle Ips sexdentatus - Prevalence, histological and ultrastructural evidence, and molecular characterization.

Ips sexdentatus (six-spined engraver beetle) from Austria and Poland were dissected and examined for the presence of pathogens. Specimens collected in Austria were found to contain the ascomycetous fungus Metschnikowia cf. typographi. Infection rates ranged from 3.6% to 26.8% at different collection sites. M. cf. typographi infected midguts were investigated by histological, ultrastructural and molecular techniques. Extraordinary ultrastructural details are shown, such as ascospores with bilateral flattened flanks resembling alar rims at both sides of their attenuating tube-like ends. These have not yet been described in other yeast species. Molecular investigations showed a close phylogenetic relationship to the fungi Metschnikowia agaves and Candida wancherniae. Presence of the entomopathogenic fungus Beauveria bassiana found in Austria was confirmed both morphologically and molecularly. The eugregarine Gregarina typographi was diagnosed most frequently. Infection rates of all I. sexdentatus specimens ranged from 21.4% to 71.9% in Austria and 54.1% to 68.8% in Poland. Other entomopathogenic protists, bacteria, or viruses were not detected.

Preliminary studies on the evolution of carbon assimilation abilities within Mucorales.

Representatives of Mucorales belong to one of the oldest lineages of terrestrial fungi. Although carbon is of fundamental importance for fungal growth and functioning, relatively little is known about enzymatic capacities of Mucorales. The evolutionary history and the variability of the capacity to metabolize different carbon sources among representatives of the order Mucorales was studied using Phenotypic Microarray Plates. The ability of 26 strains belonging to 23 nonpathogenic species of Mucorales to use 95 different carbon sources was tested. Intraspecies variability of carbon assimilation profiles was lower than interspecies variation for some selected strains. Although similarities between the phylogenetic tree and the dendrogram created from carbon source utilization data were observed, the ability of the various strains to use the analyzed substrates did not show a clear correlation with the evolutionary history of the group. Instead, carbon assimilation profiles are probably shaped by environmental conditions.

New insights on the phylogeny and biology of the fungal ant pathogen Aegeritella.

This paper evaluates the phylogenetic position of the ectoparasitic fungus Aegeritella tuberculata Balazy & Wisniewski, and broadly discusses its presence on ants in southern Poland. Field work was conducted in the Silesian Beskid Mountains in 2011-2013. The fungus was found on four species of ants: Lasius niger L., Lasius brunneus Latr., Formica lemani Bondr. and Formica fusca L. The first three species have not been noted previously in the literature as hosts of Aegeritella fungi. The infection rate ranged from 1% for Formica lemani to 21% for L. brunneus. Molecular analysis based on ITS and SSU rDNA sequences revealed close relationships between Aegeritella and Trichosporon isolates. We conclude that the genus Aegeritella-inceratae sedis until now, should be placed within the fungal group Basidiomycota, Tremellomycetes, Tremellomycetidae, Tremellales, Trichosporonaceae.

Biodegradation of nonylphenol by a novel entomopathogenic Metarhizium robertsii strain.

The biodegradation of nonylphenol (NP) by a newly isolated form of the larva fungal strain Metarhizium robertsii IM 6519 was investigated in this study. This isolate was capable of degrading 4-n-NP, and multiple metabolites were detected. The coexistence of parallel degradation pathways with versatile hydroxylation in different positions of the alkyl chain is a unique feature of this strain. Moreover, several metabolites previously described only in higher eukaryotes were detected in the fungal cultures. The degradation process led to the mineralization of 4-n-NP (with an efficiency of 36%), a great advantage of this strain that results in complete removal of toxic substrate from the environment.

Bringing Laboulbeniales into the 21st century: enhanced techniques for extraction and PCR amplification of DNA from minute ectoparasitic fungi.

Laboulbeniales is one of the most peculiar orders of Ascomycota. These fungi are characterized by an ectoparasitic life-style on arthropods, determinate growth, lack of an asexual stage, high species richness, and intractability to culture. The order Laboulbeniales, sister to Pyxidiophorales, has only recently been assigned a separate class, the Laboulbeniomycetes, based on very few ribosomal DNA sequences. So far, DNA isolations and PCR amplifications have proven difficult. Here, we provide details of isolation techniques and the application of commercially available kits that enable efficient and reliable genetic analyses of these fungi. We provide 43 newly generated Laboulbeniales ribosomal DNA sequences, among which are the first published sequences for species in the genera Gloeandromyces, Herpomyces, Laboulbenia, Monoicomyces, and Polyandromyces. DNA extractions were possible using from 1 to 30 thalli from hosts preserved in ethanol (70-100 %). In two cases, we successfully isolated DNA from thalli on dried insect collections. Laboulbeniales molecular systematics could be substantially enhanced through these improved methods by allowing more complete sampling of both taxa and gene regions.

The diversity of endophytic fungi in the above-ground tissue of two Lycopodium species in Poland.

Endophytes are a large and diverse group of fungi that colonize healthy plant tissues without causing any symptoms. The majority of studies have focused on angiosperm and conifer hosts and few have examined the endophytes of lycophytes. In the present study, we characterized culturable endophytic fungi in two closely related Lycopodium species (L. annotinum and L. clavatum) from pine, beech, oak and spruce forests across Poland. More than 400 strains were isolated but only 18 Ascomycete species were identified. Members of the Dothideomycetes dominated the fungal endophyte communities in Lycopodium. The most abundant taxa cultured were Phoma brasiliensis (from L. clavatum) and Paraconiothyrium lycopodinum (from L. annotinum). Five taxa were isolated exclusively from L. annotinum, but only two of them (Paraconiothyrium lycopodinum and Mycosphaerella sp.) were relatively abundant. Two taxa were only found in L. clavatum, namely: Stagonospora pseudovitensis and an unidentified Dothideomycete. The taxon assigned as Ascomycota 2 (SH219457.06FU) was isolated only from strobili of both host species. Direct PCR and cloning from L. annotinum shoots revealed a substantially greater endophyte richness compared with the results from culturing.

Utilization of 4-n-nonylphenol by Metarhizium sp. isolates.

Nonylphenol (4-NP) is a xenobiotic classified as an endocrine disrupting compound with an ability to interfere with hormonal systems of numerous organisms including humans. It is widely distributed not only in aquatic but also in terrestrial systems. The aim of this study was to evaluate the ability of cosmopolitan fungus Metarhizium (commonly persistent in soil as a facultative insect pathogen, controlling populations of arthropods in natural environment) to degrade 4-n-nonylphenol. All isolates examined in this work were identified to a species rank based on five, independent genetic markers. Among eight Metarhizium strains; six of them have been identified as M. robertsii, and two others as M. brunneum and M. lepidiotae. All investigated Metarhzium isolates were found to eliminate 4-n-NP with significant efficiency (initial xenobiotic concentration 50 mg L(-1)). The degradation process was very effective and at 24h of incubation 50-90% of 4-n-NP was eliminated by certain strains, while extended incubation resulted in further utilization of this compound. At the end of the experiments 64-99% of 4-n-NP was removed from the culture medium. Additionally, in all tested cultures three major metabolites were detected: 4-hydroxybenzoic acid; 2-(4-hydroxyphenyl)acetic acid and 4-hydroxyphenylpentanoic acid. The obtained results indicate that Metarhizium sp. possesses an ability to degrade NP and can serve as a potential candidate for further biodegradation studies.

Diversity of acaropathogenic fungi in Poland and other European countries.

The occurrence, species diversity and some aspects of taxonomical affinity and host selectivity of acaropathogenic fungi associated with phytophagous, saprotrophic and predacious mites in Poland and other European countries were investigated on wild and cultivated plants, in insect feeding sites under the bark and in decayed wood. From among 33 species of fungi affecting mites only five species of Entomophthorales were separated and the most numerous were Neozygites floridana mostly on Tetranychus urticae, N. abacaridis on a few eriophyid species, and Conidiobolus coronatus attacking gamasid mites most frequently of the genus Dendrolaelaps. The most frequent mite pathogens occurring in mite communities on plants and in wood infested by insects were of the genus Hirsutella. Until now 13 of their form-species have been recognized in these habitats, but only H. kirchneri, H. necatrix and H. thompsonii (including its variety synnematosa) can be treated as exclusive oligophagous pathogens of phytophagous mites, though their potential host range seems to embrace only selected eriophyid or tarsonemid mites. Taxonomical differentiation of fungal strains was based on close morphological observations and molecular analysis of ITS region sequences. Two new species of acaropathogenic fungi were described in these studies. Hirsutella danubiensis sp. nov. was found in the tetranychid T. urticae, whereas H. vandergeesti sp. nov. affected phytoseiid mites of the genera Amblyseius, Neoseiulus, Seiulus and Typhlodromus, and the tarsonemid Tarsonemus lacustris.

Laboratory trials to infect insects and nematodes by some acaropathogenic Hirsutella strains (Mycota: Clavicipitaceous anamorphs).

Laboratory assays have been carried out to artificially infect insect larvae of the birch bark-beetle (Scolytus ratzeburgi Jans.-Coleoptera, Scolytidae) and codling moth Cydia pomonella L. -Lepidoptera, Tortricidae) as well as the potato cyst nematode-Globodera rostochiensis Wollenweber, sugar beet nematode-Heterodera schachtii Schmidt and root-knot nematode-Meloidogyne hapla Chif (Nematoda, Heteroderidae), by the phialoconidia of some fungal species of the genus Hirsutella. From among four species tested on insects only H. nodulosa Petch infected about 20% of S. ratzeburgi larvae, whereas H. kirchneri (Rostrup) Minter, Brady et Hall, H. minnesotensis Chen, Liu et Chen, and H. rostrata Balazy et Wisniewski did not affect insect larvae. Only single eggs of the root-knot nematode were infected by H. minnesotensis in the laboratory trials, whereas its larvae remained unaffected. No infection cases of the potato cyst nematode (G. rostochiensis) and sugar beet nematode eggs were obtained. Comparisons of DNA-ITS-region sequences of the investigated strains with GenBank data showed no differences between H. minnesotensis isolates from the nematodes Heterodera glycines Ichinohe and from tarsonemid mites (authors' isolate). A fragment of ITS 2 with the sequence characteristic only for H. minnesotensis was selected. Two cluster analyses indicated close similarity of this species to H. thompsonii as sister clades, but the latter appeared more heterogenous. Insect and mite pathogenic species H. nodulosa localizes close to specialized aphid pathogen H. aphidis, whereas the phytophagous mite pathogens H. kirchneri and H. gregis form a separate sister clade. Hirsutella rostrata does not show remarkable relations to the establishment of aforementioned groups. Interrelated considerations on the morphology, biology and DNA sequencing of investigated Hirsutella species state their identification more precisely and facilitate the establishment of systematic positions.