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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 roztoczensissp. nov. and S. silvicolasp. nov. In addition, isolates collected from fallen shoots of Pinus sylvestris that were pruned by Tomicus sp. are described as Sporothrix tumidasp. 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.

The genus Ceratocystiopsis (Ophiostomatales, Ascomycota) includes 21 species, which can be found mainly in association with bark beetles in the Northern Hemisphere. A survey of Ceratocystiopsis species associated with bark beetles infesting Picea abies and Pinus sylvestris in Norway yielded 126 isolates, representing Ceratocystiopsis neglecta and Ceratocystiopsis rollhanseniana, and four species described herein as Ceratocystiopsis chalcographii, Ceratocystiopsis debeeria, Ceratocystiopsis norroenii and Ceratocystiopsis troendelagii. The new taxa were morphologically characterised and phylogenetically analysed on the basis of sequence data of multiple loci (ITS, LSU, beta-tubulin (TUB2), calmodulin (CAL) and translation elongation factor 1-alpha (TEF1) genes). Ceratocystiopsis norroenii and C. rollhanseniana were the most frequently isolated species, and the latter species had the wider vector range.

During surveys of insect-associated mycobiomes in Norway, Poland, and Russia, isolates with affinity to Graphilbum (Ophiostomatales, Ascomycota) were recovered. In this study, eight known Graphilbum species as well as the newly collected isolates were compared based on morphology and DNA sequence data for four gene regions. The results revealed seven new species, described here as G. acuminatum, G. carpaticum, G. curvidentis, G. furuicola, G. gorcense, G. interstitiale, and G. sexdentatum. In addition to these species, G. crescericum and G. sparsum were commonly found in Norway. All new species were recovered from conifers in association with bark beetles, cerambycid beetles, and weevils and were morphologically similar, predominantly with pesotum-like asexual morphs. Where sexual morphs were present, these were small ascomata with short necks and rod-shaped ascospores having hyaline sheaths. The results suggest that Graphilbum species are common members of the Ophiostomatales in conifer ecosystems.

Blue stain is a type of discoloration that considerably lowers the aesthetic qualities of wood. During harvesting, fungal inoculum finds its way into the wood through places of cutting, delimbing, and damage to the tree stem. The development of peripheral blue stain in the logs of Scots pine (Pinus sylvestris L.) due to damage by a harvester head was investigated, using logs felled and delimbed with a chainsaw as reference. The development of blue stain was related to the age class of the stand, the date of felling, and the microclimatic conditions of the wood storage place. The mean surface area of blue stain in logs produced with the harvester was almost 35 cm2, approximately 30% larger than that of the reference logs. In stands of the fourth age class (diameter at breast height 32 cm), the surface of blue stain was more than two times larger than that of stands of the third age class (diameter at breast height 24 cm). The mean area of blue stain in wood stored after spring felling (19.3 cm2) was about 50% of that after summer felling (38.2 cm2). In particular, the development of blue stain in logs obtained with a harvester was visible between weeks 6 and 9 after spring felling and between weeks 3 and 6 after summer felling. For both felling dates, the range of blue stain depended less on air temperature around the wood stack and depended more on the duration of storage.

The genus Ceartocystiopsis includes ascomycetes fungi belonging to the Ophiostomatales that are not well recognized in the world. Described species so far have been found mainly in association with bark beetles in the Northern Hemisphere. The aims of this study were therefore to survey of distribution of species of Ceratocystiopsis associated with bark beetles infesting Picea abies and Pinus sylvestris in Norway. Adults for 22 different bark beetle species were collected from 13 stands in Norway. During this study, we recovered 126 isolates showing affinities to Ceratocystiopsis representing six species, including two described and four undescribed taxa. The four undescribed taxa collected during this work were characterised based on their morphological characteristics and multi-gene phylogenies. Herein, we describe these new species as Ceratocystiopsis chalcographii sp. nov., Ceratocystiopsis debeeria sp. nov., Ceratocystiopsis norroenii sp. nov. and Ceratocystiopsis troendelagii sp. nov. Ceratocystiopsis norroenii and C. rollhanseniana were the most frequently isolated species although the latter species had a much wider vector range. This study expands our knowledge about the taxonomy and species diversity of Ceratocystiopsis and beetle-fungus relationships.

Fungi in the orders Ophiostomatales and Microascales (Ascomycota), often designated as ophiostomatoid fungi, are frequent associates of scolytine bark and ambrosia beetles that colonize hardwood and coniferous trees. Several species, e.g., Ophiostoma novo-ulmi, are economically damaging pathogens of trees. Because little is known regarding the ophiostomatoid fungi in Europe, we have explored the diversity of these fungi associated with hardwood-infesting beetles in Poland. This study aims to clarify the associations between fungi in the genera Ambrosiella, Graphium (Microascales), Graphilbum, Leptographium, Ophiostoma and Sporothrix (Ophiostomatales) and their beetle vectors in hardwood ecosystems. Samples associated with 18 bark and ambrosia beetle species were collected from 11 stands in Poland. Fungi were isolated from adult beetles and galleries. Isolates were identified based on morphology, DNA sequence comparisons for five gene regions (ITS, LSU, sT, TEF 1-α, and CAL) and phylogenetic analyses. In total, 36 distinct taxa were identified, including 24 known and 12 currently unknown species. Several associations between fungi and bark and ambrosia beetles were recorded for the first time. In addition, associations between Dryocoetes alni, D. villosus, Hylesinus crenatus, Ernoporus tiliae, Pteleobius vittatus and ophiostomatoid fungi were reported for the first time, and Sporothrix eucastanea was reported for the first time outside of the USA. Among the species of Ophiostomatales, 14 species were in Ophiostoma s. l., two species were in Graphilbum, nine species were in Sporothrix, and seven species were in Leptographium s. l. Among the species of Microascales, three species were in Graphium, and one was in Ambrosiella. Twenty taxa were present on the beetles and in the galleries, twelve only on beetles, and four only in galleries. Bark and ambrosia beetles from hardwoods appear to be regular vectors, with ophiostomatoid fungi present in all the beetle species. Most ophiostomatoid species had a distinct level of vector/host specificity, although Ophiostoma quercus, the most frequently encountered species, also had the greatest range of beetle vectors and tree hosts. Plant pathogenic O. novo-ulmi was found mainly in association with elm-infesting bark beetles (Scolytus multistriatus, S. scolytus, and P. vittatus) and occasionally with H. crenatus on Fraxinus excelsior and with Scolytus intricatus on Quercus robur.

Ophiostoma spp. (Ophiostomatales, Ascomycota) are well-known fungi associated with bark and ambrosia beetles (Curculionidae: Scolytinae, Platypodinae). Fungi in the Ophiostomatales include serious tree pathogens as well as agents of timber blue-stain. Although these fungi have been extensively studied in the northern hemisphere, very little is known regarding their occurrence on hardwoods in Europe. The aims of the present study were to identify and characterize new Ophiostoma spp. associated with bark and ambrosia beetles infesting hardwoods in Norway and Poland, and to resolve phylogenetic relationships of Ophiostoma spp. related to the Norwegian and Polish isolates, using multigene phylogenetic analyses. Results obtained from five gene regions (ITS, LSU, β-tubulin, calmodulin, translation elongation factor 1-α) revealed four new Ophiostoma spp. These include Ophiostoma hylesinum sp. nov., O. signatum sp. nov., and O. villosum sp. nov. that phylogenetically are positioned within the Ophiostoma ulmi complex. The other new species, Ophiostoma pseudokarelicum sp. nov. reside along with Ophiostoma karelicum in a discrete, well-supported phylogenetic group in Ophiostoma s. stricto. The results of this study clearly show that the diversity and ecology of Ophiostoma spp. on hardwoods in Europe is poorly understood and that further studies are required to enrich our knowledge about these fungi.

Acidophilic fungi are characterized by their ability to tolerate very low pH conditions and to grow optimally below pH 5. The genus Acidiella (Dothideomycetes, Capnodiales, Teratosphaeriaceae) comprises acidophilic and acidotolerant fungi that are commonly distributed in highly acidic soils, drainages and waters worldwide. This study revises Acidiella taxonomy and provides alternative DNA barcodes. The morphological characteristics were revaluated by the investigation of conidial ontogeny, and knowledge of the ecology and biogeography of the genus is summarized. The new species Acidiella polonica isolated from highly acid (pH 2.7–3.8) technogenic soil developed on mine spoils in Poland is described here. This species is distinguishable by genetic markers, morphology and its ability to grow at pH 1. Based on DNA sequence and morphological comparisons, we also show that Acidiella uranophila (syn. Fodinomyces uranophilus) is a synonym of Acidiella bohemica. Among all species, A. bohemica is the most commonly isolated and is widely distributed over four continents. Interestingly, its distribution at the global scale is limited to extremely acidic substrates (pH

Geosmithia species (Hypocreales, Ascomycota) are associates of bark beetles and other arthropods. One species, Geosmithia morbida, is a virulent tree pathogen of Juglans nigra. To date, 10 Geosmithia spp. from conifer-infesting, and at least 23 species from hardwood associated bark beetles have been reported from Europe. The aim of this study was to survey Geosmithia spp. associated with 18 bark and ambrosia beetle species in hardwood ecosystems in Poland. In addition, we evaluated the pathogenicity of the six Geosmithia species by inoculating Acer, Fagus, Quercus, Tilia and Ulmus seedlings. Our surveys yielded a total of 1060 isolates from 2915 beetles and 1887 galleries. We identified isolates using morphology and ITS, β-tubulin and TEF1-α sequences. Altogether we identified 11 species including nine previously known and two new species described here as Geosmithia fagi sp. nov. and G. pazoutovae sp. nov. In addition, a sister species G. longistipitata sp. nov., associated with Picea trees, is described here. Bark beetles from hardwoods, with exeption of Dryocoetes alni, D. villosus, Scolytus ratzeburgi and ambrosia beetles, appear to be regular vectors of Geosmithia spp. Like in other parts of the world, most Geosmithia taxa exhibited a distinct level of vector/host specificity. None of Geosmithia isolates induced any disease symptoms under the conditions of our experiment. This study highlights the need for more intensive surveys across additional areas of Central and Northern Europe, insect vectors and host tree species in order to elucidate the Geosmithia species diversity in this region.

The physiological and behavioural activities of animals have far-reaching impacts on the characteristics and functioning of soil. This includes vertebrates, which are capable of modifying the physicochemical and biochemical properties of soil. To date, however, no species is known to be responsible for the entire process of soil formation, modification and maintenance. Large-bodied birds build nests which they then use for several years or even decades. During nest construction or renovation, birds gather and transport to the nesting site organic and mineral matter that includes tree branches of various sizes, twigs, turf, straw and hay. Over time, during subsequent breeding events, adult birds supply further loads of organic matter to the nest, such as food remains, excrement, pellets, feathers, egg shells and other materials. Taking the White Stork Ciconia ciconia as an example, we have shown that the materials deposited in the nests of large-bodied birds gradually produce ornithogenic soils over the years, with distinguishable layers having different physicochemical characteristics and biochemical activities. The tested nesting substrate met the criteria for ornithogenic material; the layers had appropriate thickness and phosphorus pentoxide (P2O5) content. Results of the study indicates that the material contained in White Stork nests have the characteristics of Histosols. Moreover, such nests harbour assemblages of fungi and arthropods that contain species typical of soil mycobiota and fauna, respectively. This study is the first to describe a soil that is formed, modified and maintained entirely by vertebrates and is physically isolated from the ground. Our results highlight the fact that the nests of large birds are unique structures in ecosystems and provide a habitat for a rich and diverse assemblage of organisms.

During surveys of insect-associated mycobiomes in Norway, Poland, and Russia, isolates with affinity to

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 synnematasp. nov. and Leptographium alneumsp. 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.

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 synnematasp. nov. and Leptographium alneumsp. 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.

Dieback of young Abies alba seedlings was identified as an important limitation for natural regeneration of this species in the mountain regions of Central Europe. Approximately 2–4 weeks after emergence, during cool and wet weather, necrotic spots developed on seedling cotyledons. Later they expanded and merged to finally cover all above-ground tissues. Symptomatic seedlings of A. alba were sampled at 16 stands, and fungi were isolated from the cotyledons, stems, and roots. Morphological and molecular methods based on ITS rDNA sequencing were then used for isolate identification. In total, 105 distinct taxa were identified, with Ascomycota dominating, representing 97.4% of the isolates. The most frequently isolated taxa were Allantophomopsis lycopodina, Gyoerffyella rotula, Leptosphaeria sp., Phialocephala fortinii, Pseudaegerita sp., Scirrhia aspidiorum, Sydowia polyspora, Peyronellaea sp., Phomopsis sp., and Varicosporium elodeae. The frequency of colonization by individual fungal species differed among the three plant organs and among forest types. In symptomatic cotyledons, the dominant species were G. rotula, S. polyspora, and Pseudaegerita sp. whose pathogenicity, together with that of Gyoerffyella sp., V. elodeae, and Hormonema carpetanum, was tested on young A. alba seedlings. Only G. rotula and Gyoerffyella sp. caused serious dieback of above-ground seedling tissues and necrotic symptoms on cotyledons. This study is one of the few comprehensive reports showing that aquatic hyphomycetes commonly occur in forests beyond their preferred habitat.

Summary Cylindrocarpon-like fungi are globally distributed plant pathogens and have a wide range of host species. However, very little is known about the species that live in the topsoil of forests and their potential role in reducing the natural regeneration of tree species, particularly of forest trees that produce abundant fruit only once every few years. To enhance our understanding of the species that inhabit the topsoil, we studied the diversity and pathogenicity of Cylindrocarpon-like fungi inhabiting the litter in old-growth mixed-beech forests in the Carpathians (Poland) and in the Alps (Austria), and in a managed beech stand in the Krakowsko-Czestochowska Highland (Poland). The fungi inhabiting the beech litter were investigated using beechnuts and pine seedlings as bait. Isolates were identified based on morphology and DNA sequencing. The pathogenicity of the most common species was investigated by inoculating beech germinants. A wide range of Cylindrocarpon-like fungi were associated with the beech litter: 718 cultures representing 12 species were isolated. Five taxa were identified down to species level: namely Ilyonectria crassa, I. pseudodestructans, I. rufa, Neonectria candida and N. obtusispora, and seven species were identified to genus level (Neonectria or Ilyonectria species). Ilyonectria destructans, which is considered to be the sexual morph of ‘Cylindrocarpon destructans’, was not found. There were qualitative and quantitative differences between the different forest sites in terms of Neonectria and Ilyonectria species composition in beech litter. The isolation frequency and species richness of Cylindrocarpon-like fungi were greatest in beech litter taken from old-growth mixed-beech forests. Neonectria and Ilyonectria species were capable of killing beech germinants, suggesting that they may play a negative role in natural beech regeneration.

Fungi under the order Ophiostomatales (Ascomycota) are known to associate with various species of bark beetles (Coleoptera: Curculionidae: Scolytinae). In addition this group of fungi contains many taxa that can impart blue-stain on sapwood and some are important tree pathogens. A recent survey that focussed on the diversity of the Ophiostomatales in the forest ecosystems of the Czech Republic and Poland uncovered four putative new species. Phylogenetic analyses of four gene regions (ITS1-5.8S-ITS2 region, ß-tubulin, calmodulin, and translation elongation factor 1-α) indicated that these four species are members of the genus Ophiostoma. All four newly described species can be distinguished from each other and from closely related species based on DNA sequence comparisons, morphological characters, growth rates, and their insect associations. Based on this study four new taxa can be circumscribed and the following names are provided: Ophiostoma pityokteinis sp. nov., Ophiostoma rufum sp. nov., Ophiostoma solheimii sp. nov., and Ophiostoma taphrorychi sp. nov. O. rufum sp. nov. is a member of the Ophiostoma piceae species complex, while O. pityokteinis sp. nov. resides in a discrete lineage within Ophiostoma s. stricto. O. taphrorychi sp. nov. together with O. distortum formed a well-supported clade in Ophiostoma s. stricto close to O. pityokteinis sp. nov. O. solheimii sp. nov. groups within a currently undefined lineage A, which also includes Ophiostoma grandicarpum and Ophiostoma microsporum. This study highlights the need for more intensive surveys that should include additional countries of Central Europe, insect vectors and host tree species in order to elucidate Ophiostoma species diversity in this region. Electronic supplementary material The online version of this article (10.1007/s10482-019-01277-5) contains supplementary material, which is available to authorized users.

Ophiostoma quercus (Ascomycota, Ophiostomatales) is a globally widespread, insect-vectored fungus that colonizes a wide diversity of hardwood and conifer hosts. Although the fungus is considered to be non-pathogenic, it is closely related to the fungi that cause Dutch elm disease. We examined the global diversity of O. quercus based on a ribosomal RNA marker and three unlinked gene regions. The fungus exhibited substantial morphological diversity. In addition, O. quercus had high genetic diversity in every continent from which it was collected, although the fungus was most diverse in Eurasia. There was no evidence of geographical clustering of haplotypes based on phylogenetic and network analyses. In addition, the phylogenetic trees generated based on the different markers were non-congruent. These results suggest that O. quercus has been repeatedly moved around the globe, because of trade in wood products, and that the fungal species most likely outcrosses regularly. The high genetic diversity of the fungus, as well as its ability to utilize a wide variety of arthropod vectors and colonize a tremendous diversity of tree host species makes O. quercus truly unique among ophiostomatoid fungi.

Species of Leptographium are generally characterized by mononematous conidiophores and are commonly associated with bark beetles and weevils. These species are responsible for sapstain and in some cases serious diseases on a range of primarily coniferous trees. In comparison with coniferous trees, the occurrence of Leptographium species on hardwood trees has been poorly studied in Europe. During a survey of ophiostomatoid fungi on various tree species in Norway and Poland, three unusual species, which fit the broader morphological description of Leptographium spp., were found in association with Scolytus ratzeburgi, Dryocoetes alni and Trypodendron domesticum on a variety of hardwoods, and from wounds on Tilia cordata. Phylogenetic analyses of sequence data for three gene regions (ITS2-LSU, β-tubulin, and TEF1-α) showed that these Leptographium species are phylogenetically closely related to each other and form a well-supported lineage that included Grosmannia grandifoliae and Leptographium pruni. The first species could be distinguished from the other Leptographium species based on conidiophores arising from spiral hyphae, chlamydospore-like structures and a hyalorhinocladiella-like synanamorph in culture. The second species differs from the previous one by having distinctly shorter conidiophores and smaller conidia. This species also produces a well-developed sporothrix-like synanamorph with denticulate conidiogenous cells. Based on these unusual morphological characteristics and distinct DNA sequences, these fungi were recognised as new taxa for which the names Leptographium trypodendri sp. nov. and L. betulae sp. nov. are provided. The third group of isolates belonged to Grosmannia grandifoliae, representing the first report of this species outside of the USA. The newly defined G. grandifoliae complex is the first species complex in Leptographium s.l. consisting of only hardwood-infecting species.

Bark beetles (Coleoptera, Scolytinae) are known to be associated with fungi, particularly species of the orders Ophiostomatales and Microascales. However, very little is known about other ectosymbionts of phloeophagous bark beetles on Pinaceae. In this study, we examined the Geosmithia species associated with eight bark beetle species infesting Picea abies and Pinus sylvestris branches in Poland. Fungi were isolated from 1 731 samples collected from 14 study sites. We identified a total of 653 isolates that were sorted into nine taxa based on their phenotypic similarity and phylogeny of their ITS-LSU regions of rDNA, β-tubulin, elongation factor 1α and the second-largest subunit of the RNA polymerase II gene. They represented nine species without formal names. There were large quantitative and qualitative differences in the composition of Geosmithia communities between P. sylvestris and P. abies trees. The proportion of samples infested with Geosmithia species suggests that this association is more widespread among bark beetles infesting branches of P. sylvestris . In addition, these beetles were vectors of different Geosmithia species compared with than the beetles that colonize P. abies . In mixed-conifer forests, the Geosmithia communities were more diverse and richer than in pure spruce or pine stands, where the insects Pityogenes chalcographus and Pityophthorus pityographus with low host-specificity play a distributing role for various Geosmithia species. Among eight bark beetle species examined, only P. bidentatus , P. pityographus , P. chalcographus and Polygraphus poligraphus acted as effective vectors for Geosmithia species. The following hypothesis emerges from these studies: changes in the composition of ectosymbionts of pine- and spruce-infesting bark beetles in Central Europe run along a gradient of thickness of the wood substrata preferred by insects.

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.

Fungi from the genus Geosmithia (Ascomycota: Hypocreales) are associated with bark beetles (Coleoptera: Scolytinae), though little is known about ecology, diversity, and distribution of these fungi across beetle and its host tree species. This study surveyed the diversity, distribution and vector affinity of Geosmithia isolated from subcortical insects that colonized trees from the family Pinaceae in Central and Northeastern Europe. Twelve Geosmithia species were isolated from 85 plant samples associated with 23 subcortical insect species (including 14 bark beetle species). Geosmithia community composition was similar across different localities and vector species; although the fungal communities associated with insects that colonized Pinus differed from that colonizing other tree species (Abies, Larix, and Picea). Ten Geosmithia species from four independent phylogenetic lineages were not reported previously from vectors feeding on other plant families and seem to be restricted to the vectors from Pinaceae only. We conclude that presence of such substrate specificity suggests a long and stable association between Geosmithia and bark beetles.

Bark weevils are consistently associated with various fungi. They act as effective vectors for root-rot fungi, the rust pathogen and the ophiostomatoid fungi. In comparison with bark beetles, the interaction between ophiostomatoid fungi and bark weevils has been poorly studied in Europe. This study aims to clarify the ties among ophiostomatoid fungi and their weevil vectors in Pinus sylvestris. Samples associated with three bark weevils, including Pissodes castaneus, P. piniphilus and P. pini were collected from seven pine stands in Poland. Fungi were isolated from laboratory-reared and field-collected beetles, larvae and galleries of weevils. Isolates were identified based on morphology, DNA sequence comparison for two gene regions (ITS, s-tubulin) and phylogenetic analyses. Fourteen morphological and phylogenetic ophiostomatoid species were identified among the 1,219 isolates. These 14 species included 11 novel associations between fungi and bark weevils. The most commonly encountered fungal associates of bark Pissodes species were Leptographium procerum, Ophiostoma quercus and O. minus. The spectrum of fungal associates was similar in three bark weevil species, despite some differences between species. Except for P. castaneus, all species of bark weevil were frequently associated with ophiostomatoid fungi. Pissodes castaneus does appear to be a consistent vector of L. procerum and Sporothrix inflata while O. minus, O. quercus and O. cf. rectangulosporium were often found in association with P. piniphilus and P. pini.

Bark beetles are known to be associated with fungi, especially the ophiostomatoid fungi. However, very little is known about role of pine weevils, e.g., Hylobius abietis, as a vector of these fungi in Europe. The aims of our study were to demonstrate the effectiveness of H. abietis as a vector of ophiostomatoid fungi in Poland and to identify these fungi in Scots pine seedlings damaged by weevil maturation feeding. Insects and damaged Scots pine seedlings were collected from 21 reforestation sites in Poland. The fungi were identified based on morphology, DNA sequence comparisons for two gene regions (ITS, β-tubulin) and phylogenetic analyses. Sixteen of the ophiostomatoid species were isolated and identified. In all insect populations, Leptographium procerum was the most commonly isolated fungus (84 %). Ophiostoma quercus was also found at a relatively high frequency (16 %). Other ophiostomatoid fungi were found only rarely. Among these rarer fungi, four species, Leptographium lundbergii, Ophiostoma floccosum, Ophiostoma piliferum and Sporothrix inflata, were isolated above 3 %. L. procerum was isolated most frequently and was found in 88 % of the damaged seedlings. S. inflata was isolated from 26 %, while O. quercus occurred in 10 % of the seedlings. This study confirmed that L. procerum and O. quercus were common associates of H. abietis, while others species were found inconsistently and in low numbers, indicating causal associations. H. abietis also acted as an effective vector transmitting ophiostomatoid species, especially L. procerum and S. inflata, to Scots pine seedlings.

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.

Appropriate management of invasive fungi requires adequate understanding of their global diversities and movement histories. The fungus Leptographium procerum is associated with root-colonizing forest insects in pine forests throughout the world, and may have contributed to the aggressive behaviour of the red turpentine beetle (Dendroctonus valens) in the beetle's invasive range in China. We used microsatellites and mating type loci to investigate the global diversity of L. procerum and the source population of L. procerum associated with D. valens in China. Clustering analyses supported the separation of the fungal data set into three genetically and geographically-distinct clusters: Europe, North America, and China. The fungus had the highest genetic diversity in Europe, followed by North America and China. Analyses using Approximate Bayesian Computation supported Europe as the most likely source of the North American and Chinese populations. Overall, the results suggested that Europe is the global centre of diversity of L. procerum. Furthermore, they suggested that L. procerum most likely arrived in China independently of D. valens and adopted this beetle as a vector after its introduction.

Summary Ophiostomatoid fungi are carried by various bark beetles. However, very little is known about the role of these fungi in conifer roots. We studied ophiostomatoid fungi in roots of dying and dead Pinus sylvestris trees and tested the potential phytotoxicity of some isolates using a sensitive bioassay with Lepidium sativum in Poland. Fungi were identified based on their morphology and DNA sequencing. Three ophiostomatoid fungi, Leptographium procerum, Sporothrix inflata and Ophiostoma pallidulum, were isolated from the roots. The most abundant soil-borne fungus, S. inflata, and relatively rare O. pallidulum were isolated for the first time from roots of dying and dead pine trees. The frequency of S. inflata and O. pallidulum correlated with tree decline. The fungi were isolated more frequently from roots of dead than dying trees. Sporothrix inflata and O. pallidulum slightly reduced the stem and root growth of L. sativum. Leptographium procerum reduced more significantly root than stem growth. This species reduced root elongation 32–54% after 10–17 days of incubation.

Two species of blue-stain fungi with similar morphologies, Ophiostoma brunneo-ciliatum and Ophiostoma clavatum, are associates of bark beetles infesting Pinus spp. in Europe. This has raised questions whether they represent distinct taxa. Absence of herbarium specimens and contaminated or mistakenly identified cultures of O. brunneo-ciliatum and O. clavatum have accentuated the uncertainty regarding their correct identification. The aim of this study was to reconsider the identity of European isolates reported as O. brunneo-ciliatum and O. clavatum by applying DNA-based identification methods, and to provide appropriate type specimens for them. Phylogenetic analyses of the ITS, βT, TEF-1α and CAL gene sequences revealed that the investigated isolates represent a complex of seven cryptic species. The study confirmed that ITS data is insufficient to delineate species in some Ophiostoma species clusters. Lectotypes and epitypes were designated for O. clavatum and O. brunneo-ciliatum, and three new species, Ophiostoma brunneolum, Ophiostoma macroclavatum and Ophiostoma pseudocatenulatum, are described in the newly defined O. clavatum-complex. The other two species included in the complex are Ophiostoma ainoae and Ophiostoma tapionis. The results suggest co-evolution of these fungi in association with specific bark beetles. The results also confirm the identity of the fungus associated with the pine bark beetle Ips acuminatus as O. clavatum, while O. brunneo-ciliatum appears to be mainly associated with another pine bark beetle, Ips sexdentatus.

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.

Interakcje pomiędzy kalusem trzech genotypówAbies albaa grzybami o różnym statusie ekologicznym

Leptographium piriforme Greif, Gibas & Currah, described in 2006 from western Canada has recently been collected in Jaroszowiec in Poland. The ophiostomatoid fungus L. piriforme was isolated from fallen Pinus sylvestris L. shoots infested by Tomicus spp. Isolates were identified to species using morphological characteristics and nuclear ribosomal DNA sequences. Two-year-old P. sylvestris seedlings were wound-inoculated with one isolate of L. piriforme. Leptographium piriforme produced necrotic lesions in the phloem of the seedlings, but none of the plant inoculated with this fungus died. This is the first report of L. piriforme from Poland and entire Europe.

Summary The aim of this study was to identify fungi associated with Cryphalus piceae on European silver fir(Abies alba) in Poland and to test the pathogenicity of selected isolates. Fungi were isolated from fivepopulations of overwintered adults and their galleries. A great diversity of taxa was associated withC. piceae. In total, 2487 isolates, including 58 species distributed in 25 genera, were obtained. The twomost frequently isolated fungi, an undescribed species of the genus Geosmithia and Ophiostomapiceae,appeared to be specifically associated with C. piceae, whereas Pesotum fragrans, Pesotum sp. andSporothrix sp. were sporadically associated. Two-year-old seedlings of silver fir were wound-inoculated with three species of fungi (Geosmithia sp., O. piceae and Pesotum sp.) recovered fromC. piceae. Only Pesotum sp. showed pathogenic ability, but we do not consider it to be an importantpathogen of A. alba. 1 Introduction The European silver fir (Abies alba Mill.) is a native species of the mountains of southernand central Europe. It is also found in Belarus, Asia and the Caucasus Mountains. InPoland, silver fir occurs in the Sudeten, the Carpathian and the S´wie˛tokrzyskie Mountains,but only forms pure stands in the eastern Carpathians (Boratyn´ski 1983). For many years,a gradual decline in its vigour has been observed in the Polish mountains, particularly in theSudeten Mountains (Bernadzki 1983). Recent studies have shown that, although abundantregeneration of the silver fir occurs in these areas (Jaworski et al. 1995; Filipiak andUfnalski 2004), significant tree damage is present. The fir stands, weakened by manyabiotic, biotic and anthropogenic factors, are attacked by phloem-feeding insects. Amongstthe bark beetles, which are secondary pests in these forests, considerable economic damageis caused by the silver fir bark beetle Cryphalus piceae (Ratz.) (Coleoptera: Scolytidae)(Michalski and Mazur 1999).Cryphalus piceae occurs in Europe and Asia, affecting several species of fir (Abies), buthas also been recorded from species of Pinus, Picea and Larix. It is usually found in themountainous regions of Poland where A. alba occurs. The fir bark beetle has a singlegeneration per year in mountain areas. Adults fly in May and June, and larvae develop inthe summer. Cryphalus piceae mainly attacks the branches and trunks of weakened, dyingand wind-fallen trees and speeds up the dieback process in weakened fir stands (Michalskiand Mazur 1999).Many bark beetle species carry ophiostomatoid fungi capable of causing blue-stain infreshly cut logs and, sometimes, damage to living trees. In some cases, the relationship isextremely close, as between Ips sexdentatus (Bo¨rn.) and Ophiostoma ips (Rumb.) Nannf.For. Path. 40 (2010) 133–144 doi: 10.1111/j.1439-0329.2009.00620.x 2009 Blackwell Verlag GmbH

Bark beetles (Coleoptera: Scolytidae) infesting Norway spruce trees are known to be associated with fungi, especially species of Ophiostoma sensu lato and Ceratocystis. However, very little is known about these fungi in Poland. In this study, we examined the ophiostomatoid species associated with seven species of bark beetles developing in Norway spruce (Picea abies). Fungi were isolated from the beetles and their galleries at ten sites in southern Poland. We identified a total of 2,769 fungal isolates that belong to 19 ophiostomatoid species. These 19 species included a total of 16 associations between fungi and bark beetles that had not been previously recorded. The isolated fungal species were similar to those previously reported. The species spectrum and relative abundance of fungal associates were similar in seven bark beetle species, despite some differences between species. Except for Pityophthorus pityographus, all bark beetles species were frequently associated with ophiostomatoid fungi. The most commonly encountered fungal associates of bark beetles were the following: Ophiostoma ainoae, O. bicolor, O. piceae sensu lato and Grosmannia piceiperda. The results support the hypothesis that pathogenic ophiostomatoid fungi are important for aggressive bark beetles to kill trees. The most virulent fungal associate of bark beetles, Ceratocystis polonica was most commonly associated with Ips typographus. The most frequent ophiostomatoid species isolated from the beetles and galleries of P. pityographus were O. ainoae and Graphium fimbriisporum.

Wstępne badania nad patogenicznością trzech grzybów siniznowych związanych zTetropiumspp. na świerku pospolitym w Polsce

The study identified and measured frequency of fungal species associated with Tomicus minor (Hart.) on Pinus sylvestris L. (Scots pine) in Poland. Additionally, fungal succession in P. sylvestris sapwood was investigated during a 12 week period following an attack by this insect. Fungi were isolated from five populations of overwintered adult beetles and their galleries with 59 species of fungi being represented among the 2880 cultures obtained. The most frequent species, Ophiostoma canum (Münch) Syd. & P. Syd., Hormonema dematioides Lagerb. & Melin, and Ambrosiella tingens (Lagerb. & Melin) L.R. Batra, appeared to be specifically associated with T. minor. The succesional changes in species composition during a 12 week period following an attack by T. minor were observed. The pattern of fungal succession in P. sylvestris sapwood essentially agreed with a general scheme of fungal succession in tree sapwood infested by bark beetles. Ambrosiella tingens was the first invader of sapwood and occurred most frequently in its deeper layers. Ophiostoma canum, H. dematioides, and other molds were also often isolated from the sapwood; however, they were most common at a depth of 5 mm during the initial phase of fungal colonization. Later, Ophiostoma canum followed A. tingens in the sapwood invasion.

Summary The association between Pityogenes bidentatus and fungi was studied in young, managed Pinus sylvestris stands in Poland. Fungi were isolated from emerged adults and their galleries collected from four populations. In total, 2089 fungal isolates including 42 species, were obtained. Penicillium sp. 1 and Geosmithia sp. 1 were the most commonly isolated fungi from beetles (49% and 41% of beetles respectively). Geosmithia sp. 1 species was the dominant species in P. bidentatus galleries with a frequency of occurrence of 57.9%. Hormonema dematioides was the second most abundant fungus in gallery systems (17.1% of wood samples). Two of the isolated Geosmithia species were previously undescribed. Pityogenes bidentatus also vectored three ophiostomatoid species: Ophiostoma minus, O. piceae and Graphium sp. ‘W’. These species were occasionally isolated from beetles and their galleries, suggesting a non-specific relationship.

The species composition of fungi associated with Ips cembrae was studied in the Raciborskie forests, Poland. The fungi were isolated from overwintered adults, larvae, new adults and from galleries at various stages of development. The results showed that there was a great diversity of fungi associated with this insect. We isolated 2877 cultures, including 61 species. The most important group of fungi were ophiostomatoid species. From these, Ceratocystis laricicola, Ophiostoma brunneo-ciliatum and Graphium laricis were commonly detected, whereas eight other species occurred less frequently or sporadically. Ophiostoma ips is documented here as a new associate of I. cembrae. Ceratocystis laricicola was shown to be the primary invader occurring most frequently at early stages of brood development, particularly in the sapwood. Ophiostoma brunneo-ciliatum and G. laricis were secondary invaders following C. laricicola. In the later stages of brood development other ophiostomatoid fungi appeared.

The association between Tomicus piniperda L. (Col., Scolytinae) and fungi was studied in a Pinus sylvestris L. forest in Mielec-Mościska. Fungi were isolated from overwintered adult beetles taken from two stands situated in different distance from timber yard. Two media were used for isolation. The results showed great diversity of fungi associated with T. piniperda: 1895 cultures, representing 64 species, were isolated. Penicillia and Hormonema dematioides were the dominant species, found in 20.2% and 17.8% of all beetles, respectively. A frequently isolated ophiostomatoid fungi was Ophiostoma minus. Qualitative and quantitative differences in the mycobiota composition of this insect between two stands were detected. The highest richness and diversity of fungal species appeared in the samples taken from the location where the trees were heavily damaged by shoot-feeding of T. piniperda. Differences were most clear for the pathogenic O. minus, which was a common fungal associate of the insects in this stand.

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.

The species composition and virulence of fungi associated with Tomicus piniperda were studied at eight locations in Poland. The fungi were isolated from phloem and sapwood samples taken from insect galleries and then identified. The virulence of the most common ophiostomatoid species was evaluated through inoculations using two-year-old Scots pine seedlings. A great diversity of fungi were found associated with T. piniperda, including 4 837 cultures and 67 species. The most important groups of fungi were the ophiostomatoids and moulds, including mainly Penicillium, Trichoderma and Mucor species. Among ophiostomatoid fungi, Ophiostoma minus and O. piceae dominated, with a frequency of occurrence of 32.4 and 11.5% of inspected galleries, respectively. Occasionally isolated species included Leptographium lundbergii, L. procerum, L. wingfieldii, Graphium pycnocephalum and Graphium sp. 'W'. In general, the frequency of the ophiostomatoid species was highly variable among locations. Leptographium wingfieldii and O. minus, were the only species capable of killing whole plants and penetrated deeper into the sapwood than other species (87-100% mortality during the 11 week incubation period). Other fungi, including O. piliferum, O. piceae and L. procerum, were considerably less virulent. associated fungi / Leptographium / Ophiostoma / Pinus sylvestris / Tomicus piniperda / virulence