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Ophiostomatales (Ascomycota) contains many species, most of which are associated with bark beetles. Some members of this order are plant or animal pathogens, while others colonize soil, different plant tissues, or even carpophores of some Basidiomycota. However, little is known about soil-inhabiting Ophiostomatales fungi. A survey of these fungi associated with soil under beech, oak, pine, and spruce stands in Poland yielded 623 isolates, representing 10 species: Heinzbutinia grandicarpa, Leptographium procerum, L. radiaticola, Ophiostoma piliferum, O. quercus, Sporothrix brunneoviolacea, S. dentifunda, S. eucastaneae, and two newly described taxa, namely Sporothrix roztoczensis sp. nov. and S. silvicola sp. nov. In addition, isolates collected from fallen shoots of Pinus sylvestris that were pruned by Tomicus sp. are described as Sporothrix tumida sp. nov. The new taxa were morphologically characterized and phylogenetically analyzed based on multi-loci sequence data (ITS, β-tubulin, calmodulin, and translation elongation factor 1-α genes). The Ophiostomatales species were especially abundant in soil under pine and oak stands. Leptographium procerum, S. silvicola, and S. roztoczensis were the most frequently isolated species from soil under pine stands, while S. brunneoviolacea was the most abundant in soil under oak stands. The results highlight that forest soil in Poland has a wide diversity of Ophiostomatales taxa, but further studies are required to uncover the molecular diversity and phylogenetic relationships of these fungi, as well as their roles in soil fungal communities.

Sporothrix (Sordariales, Ascomycota) is a well-supported monophyletic lineage within the Ophiostomatales, species of which occur in a diverse range of habitats including on forest trees, in the soil, associated with bark beetles and mites as well as on the fruiting bodies of some Basidiomycota. Several species have also been reported as important human and animal pathogens. During surveys of insect- and wound-associated Ophiostomatales from hardwood trees in Poland, many isolates with affinity to Sporothrix were recovered. In the present study, six undescribed Sporothrix spp. collected during these surveys are characterized based on their morphological characteristics and multi-locus phylogenenetic inference. They are described as Sporothrix cavum, Sporothrix cracoviensis, S. cryptarchum, S. fraxini, S. resoviensis, and S. undulata. Two of the Sporothrix spp. reside in the S. gossypina-complex, while one forms part of the S. stenoceras-complex. One Sporothrix sp. is a member of lineage F, and two other species grouped outside any of the currently defined species complexes. All the newly described species were recovered from hardwood habitats in association with sub-cortical insects, wounds or woodpecker cavities. These species were morphologically similar, with predominantly asexual states having hyaline or lightly pigmented conidia, which produce holoblastically on denticulate conidiogenous cells. Five of the new taxa produce ascomata with necks terminating in long ostiolar hyphae and allantoid ascospores without sheaths. The results suggest that Sporothrix species are common members of the Ophiostomatales in hardwood ecosystems of Poland.

Bark beetles belonging to the genus Dryocoetes (Coleoptera, Curculionidae, Scolytinae) are known vectors of fungi, such as the pathogenic species Grosmannia dryocoetidis involved in alpine fir (Abies lasiocarpa) mortality. Associations between hardwood-infesting Dryocoetes species and fungi in Europe have received very little research attention. Ectosymbiotic fungi residing in Ceratocystiopsis and Leptographium (Ophiostomatales, Sordariomycetes, Ascomycota) were commonly detected in previous surveys of the Dryocoetes alni-associated mycobiome in Poland. The aim of this study was to accurately identify these isolates and to provide descriptions of the new species. The identification was conducted based on morphology and DNA sequence data for six loci (ITS1-5.8S, ITS2-28S, ACT, CAL, TUB2, and TEF1-α). This revealed two new species, described here as Ceratocystiopsis synnemata sp. nov. and Leptographium alneum sp. nov. The host trees for the new species included Alnus incana and Populus tremula. Ceratocystiopsis synnemata can be distinguished from its closely related species, C. pallidobrunnea, based on conidia morphology and conidiophores that aggregate in loosely arranged synnemata. Leptographium alneum is closely related to Grosmannia crassivaginata and differs from this species in having a larger ascomatal neck, and the presence of larger club-shaped cells.

Relict charcoal hearths (RCHs) increases soil fertility in forest ecosystems. However, the effects of RCHs on the activity and abundance of soil microorganisms remain unknown. In this paper, we analysed the impact of relict charcoal production on the soil enzymatic activity and composition of soil bacterial and fungal communities in Scots pine forests of the Manowo Forest District in northern Poland. Moreover, we determined the effect of relict charcoal production on the soil properties. Our research was conducted by comparing the physical, chemical, enzymatic and microbiological properties of charcoal-enriched and charcoal-free soils. Significant differences in physical properties were found between these two soil types in terms of their structure and water holding capacity. As expected, horizons enriched with charcoal were characterised by a significantly higher organic carbon content (4.7% on average compared to 2.2% in control horizons), and also by a considerably higher content of available phosphorus (an average of 64.07 mg·kg−1 compared to 36.21 mg·kg−1 in the control). Similarly, RCH horizons displayed a higher pH and higher contents of Ca and Na cations. These results indicated that RCH soils provided more favourable conditions for the soil microbiome, as reflected by the higher enzymatic activity and diversity of the microorganisms. Moreover, bacterial and fungal communities in RCH soils were more diverse and had greater species/genera richness, especially in the case of fungi. Members of the genus Rhodoplanes dominated the bacterial community at both RCH and non-RCH sites, followed by Streptomyces, Burkholderia, Skermanella, Tsukamurella and Candidatus Solibacter. Both culture- and next generation sequencing (NGS)-based analyses showed that soil fungal communities were dominated by Ascomycota, with Penicillium as the most abundant genus. Our results showed that hearth soils may represent a significant C pool in the forest ecosystem. This study supports the strategy of safeguarding such charcoal-enriched soils as precious C reservoirs and ecologically important biodiversity hotspots. Moreover, the application of charcoal may effectively increase the microbial diversity of forest soils, especially during the reforestation or re-cultivation of disturbed habitats.

Members of the genus Fusarium and related genera are important components of many ecosystems worldwide and are responsible for many plant diseases. However, the structure of beech litter-inhabiting Fusarium communities and their potential role in reducing the natural regeneration of European beech are not well understood. To address this issue, we examined Fusarium communities in the litter of uneven-aged, old-growth beech-dominated forests in the Carpathians (Poland) and in the Alps (Austria), and in a managed beech stand (Poland). The fungi inhabiting beech litter were investigated using beechnuts and pine seedlings as bait. The pathogenicity of the most common species was investigated by inoculating beech germinants. Fusarium spp. were identified based on morphology and DNA sequence comparisons of RPB2 and TEF1-α genes, combined with phylogenetic analyses. Twelve fungal species were identified from 402 isolates, including nine known and three currently undescribed species. The isolates resided in three species complexes within the genus Fusarium. These were the F. oxysporum (one taxon), F. sambucinum (three taxa), and F. tricinctum (six taxa) species complexes. In addition, one isolate was assigned to the genus Neocosmospora, and one isolate could be placed within the genus Fusicolla. The most frequently isolated fungi from beechnuts and beech germinants were F. avenaceum (Fr.) Sacc., F. sporotrichioides Sherb. and Fusarium sp. B. The structure and abundance of species within Fusarium communities varied by beech forest type. The species richness of Fusarium spp. was greatest in old-growth beech-dominated stands, while abundances of Fusarium spp. were higher in managed beech-dominated stands. Pathogenicity tests showed that all four Fusarium species isolated from beechnuts and beech germinants could cause germinants to rot beech, suggesting that these fungi may play a negative role in the natural beech regeneration.

Globally, tree-holes are important ecological component of forest and woodlands. Numerous microorganisms rely on cavities, both natural and those excavated by primary cavity nesting birds, mainly by woodpeckers, for their survival and reproduction. However, the fungi occurring in cavities are not well characterized. Specifically, very little is known about the fungal communities inhabiting the woodpecker nest cavities. Therefore, in this study, we investigated the fungal diversity of cavities in southern Poland. The samples were collected from freshly excavated woodpecker nest cavities using a nondestructive method (ND). The spatial distribution of fungal communities within the cavities was evaluated by sampling different parts of a single cavity using a destructive method (D). We detected 598 fungal isolates that included 64 species in three phyla and 16 orders using the ND method. Most of the fungi isolated from the cavities represented the phylum Ascomycota (73.9% of the isolates) with 11 orders, and Microascales was the predominant order (30% of the isolates). The most common species detected was Petriella musispora, which was isolated from 65% of the cavities. A total of 150 isolates (25%) were members of Basidiomycota, with Hymenochaetales being the dominant order (16% of the isolates). The basidiomycetous fungi were isolated from 55% of the cavities. Several taxa closely related to the pathogenic fungi and associated with secondary animal infections were detected in the wood of cavities. We identified different fungal communities in the three cavity parts using the D method. The cavity entrance had more number of species than the middle and bottom parts. The results of this study advanced our current knowledge on the mycobiota in woodpecker nest cavities and provided preliminary evidence for tree cavities being the hotspot for fungal diversity.

Geosmithia species (Ascomycota: Hypocreales) are common ectosymbionts of scolytine bark and ambrosia beetles that feed on coniferous and deciduous trees in different forest ecosystems. Geosmithia morbida is the canker pathogen that causes extensive mortality of Juglans nigra. Because little is known regarding the Geosmithia species on European silver fir (Abies alba), we have investigated the diversity and abundance of these fungi associated with insects infesting European silver fir in Poland. Samples associated with eight beetle species were collected from three fir forests. Fungi were isolated from beetles and galleries. Isolates were identified based on morphological characteristics, DNA sequence comparison for three gene regions (ITS, ßT, TEF1-α), and phylogenetic analyses. Geosmithia was detected in 33% of the total 531 beetle samples obtained from A. alba. Two undescribed species of Geosmithia were distinguished, Geosmithia sp. 9 and Geosmithia sp. 16. Associations of Pityokteines spp. with Geosmithia fungi were recorded for the first time. Pityokteines vorontzowi and Pityophthorus pityographus appear to be regular vectors for Geosmithia sp. 9 and Geosmithia sp. 16, respectively. Pityokteines curvidens and Cryphalus piceae were associated with Geosmitha sp. 9 at lower frequencies.

The purpose of this study was to determine the species composition of communities of fungi inhabiting stumps of Pinus nigra depending on the period of their exposure. After 6 and 18 months of stump exposure two species of Basidiomycota distinctly dominaled: Stereum sanguinolentum and Phanerochaete gigantea. After 30 months of exposure the communities of fungi were the most diversified. A longer period of stump exposure resulted in the decrease of numbers of isolates of Ph. gigantea and S. sanguinolentum. However, other species belonging to Basidiomycota increased their numbers. As the wood decompositionn progressed, however, the fungi belonging to Deuteromycota were isolated more often.

The mycobiota of the beetles of the phloem-feeding spruce bark beetle, Ips typographus was studied. The most important group of fungi were the ophiostomatoid fungi. Among them O. penicillatum was very frequent ophiostomatoid species. Other common fungi were O. ainoae, O. bicolor, O. piceaperdum and O. piceae. The ophiostomatoid fungi were often more frequent in beetles collected in galleries than in the beetles caught With a trap. Generally the ophiostomatoid fungi were more ofien isolated from the beetle.s bathed in sterile water for 30 seconds. However C. polonica, O. ainoae, and O. minutum occurred most abundantly in the beetles disinfected in 96% ethyl alcohol for 15 and 30 seconds.

Ophiostomatoid fungi are known to be associated with Tomicus spp. on Pinus sylvestris. However, very little is actually known about the fungi present in the pine shoots damaged by these insects. The aim of this study was to survey species of Ophiostoma s.lato associated with fallen shoots of Scots pine pruned by Tomicus spp. in Poland. The study was conducted in four pure Scots pine stands in southern and south-western part of the country. Fungi were identified based on morphology and DNA sequence comparison for two gene (the ITS rDNA region and ß–tubulin). In total, 64 isolates obtained, represent seven species of ophiostomatoid fungi, including their asexual states of the genera. Two of them, Ophiostoma sp. 1 and Sporothrix sp. 1 probably represent new taxa. All species were found at very low frequencies. Among them, Ophiostoma minus, Ophiostoma sp. 1 and Sporothrix sp. 1 were the most frequently isolated, with a frequency of 2.0%. Occasionally, isolated species included Leptographium piriforme, Ophiostoma canum, Ophiostoma floccosum and Grosmannia cucullata-like. Association of species of Ophiostoma s. lato with Tomicus spp. and the taxonomic status of two new species are discussed.

The pine-infesting weevils are known to be effective vectors of ophiostomatoid fungi. To understand more about fungal virulence of these fungi, inoculation studies were conducted on Scots pine (Pinus sylvestris L.). Two-year-old seedlings were wound-inoculated with one of eleven ophiostomatoid fungi associated with pine-infesting weevils. After 11 weeks, a darkened lesion, extending from the point of inoculation, was observed in all species, except for Ophiostoma cf. abietinum Marm. & Butin, Ophiostoma quercus (Georgev.) Nannf., and Sporothrix inflata de Hoog. Seedling mortality was observed in seedlings inoculated with Leptographium truncatum (M.J. Wingf. & Marasas) M.J. Wingf., Leptographium lundbergii Lagerb. & Melin, Leptographium procerum (W.B. Kendr.) M.J. Wingf., Grosmannia radiaticola (J.J. Kim, Seifert & G.H. Kim) Zipfel, Z.W. de Beer & M.J. Wingf., Ophiostoma floccosum Math.-Käärik, Ophiostoma minus (Hedgc.) Syd. & P. Syd., and Ophiostoma piliferum (Fr.) Syd. & P. Syd. Ophiostoma minus and L. truncatum caused the largest lesions and sapwood blue-stain in Scots pine. Grosmannia radiaticola, Ophiostoma piceae (Münch) Syd. & P. Syd., O. floccosum, O. piliferum, L. lundbergii,and L. procerum produced significantly smaller lesions and sapwood blue-stain than O. minus and L. truncatum, while O. cf. abietinum, O. quercus and S. inflata did not cause any lesions.

This study dealt with the species distribution and frequency of ophiostomatoid fungi associated with the bark beetle Ips typographus on Norway spruce and Scots pine in north-eastern Poland. At all locations high spruce bark beetle damage has occurred in 2002-2003. Fungi were isolated from beetles and from brood systems of trees infested by the spruce bark beetle. The ophiostomatoid fungi were represented by 13 species. A similar spectrum of ophiostomatoid fungi as that recorded from Picea abies was associated with I. typographus on Pinus sylvestris trees. The most frequent ophiostomatoid species isolated from beetles, phloem and sapwood of Norway spruce were O. bicolor and O. penicillatum. The frequency of occurrence of ophiostomatoid fungi varied significantly among the examined locations. O. bicolor was the most frequently found species on Scots pine infested by I. typographus. The potential role of ophiostomatoid fungi in the epidemiology of I. typographus is discussed. Additionally, we also recorded how the ophiostomatoid fungi associated with spruce bark beetle could grow into phloem and sapwood of Pinus sylvestris trees.

The assemblage and abundance of fungal species associated with Hylurgops palliatus on Pinus sylvestris, were studied in Poland. Fungi were isolated from beetles of H. palliatus and their galleries collected from three populations. In total, 1832 fungal isolates, including 38 species, were obtained. The most important group of fungi were the ophiostomatoid fungi. Among the seven species of ophiostomatoid fungi isolated, Leptographium lundbergii and an unknown species of Graphium sp. code-named "W" were the dominant species. Ophiostoma minus and O. piceae were also relatively common. All ophiostomatoid species, except L. lundbergii occurred more frequently on the overwintered adults than in gallery systems. Ophiostoma minus and O. piliferum are documented here as new associates of H. palliatus.

Phytopythium and Pythium species are known to be soil-born oomycete pathogens of forest trees in Europe. Little is known, however, about the presence of these micro-organisms in Polish oak forests. During the period 2007–2009 a comprehensive study of Phytophthora species in soils of oak forests in southern Poland was conducted using baiting technique. In this study, baits were also colonized by oomycete resembling Pythium species. Based on morphological characteristics and the ITS sequences comparisons, 10 species of Phytopythium and Pythium were isolated from the soil-root samples, including three putative new species belonging to the genus of Phytopythium. The most commonly encountered Pythium species was Pythium anandrum. The present study demonstrates for the first time that Phytopythium citrinum and Pythium diclinum can also act as soil-borne organisms in oak forests. In addition, these species were reported for the first time in Poland.