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The aim of the presented study was to compare the diversity and composition of fungal communities associated with the roots and the rhizosphere soil of P. abies and Larix sp. in mid-age and mature managed forest stands in Lithuania. We also aimed to assess the presence of fungi–host-specific associations, i.e., whether Larix sp. stands could provide habitats for soil fungi currently associated with P. abies. The study sites were 10 Larix sp. and 10 P. abies forest stands in Lithuania. For the study, 100 root samples and 10 organic and 10 mineral soil samples were collected in P. abies stands as well as the same number in Larix sp. stands, and DNA was isolated, amplified using ITS2 rDNA as a marker and subjected to high-throughput sequencing. The results showed that the Shannon diversity index of fungal communities was similar between the two tree species when compared either between root (H = 4.26 P. abies and H = 3.82 Larix sp.), organic soil (H = 5.12 P. abies and H = 5.13 Larix sp.) or mineral soil (H = 4.71 P. abies and H = 4.29 Larix sp.) samples. Multivariate analysis showed that the fungal community composition in the organic and mineral soil samples of both P. abies and Larix sp. were similar, and thus, overlapping. The analysis also showed that the distribution of fungal species was denser in the roots and organic soil but more scattered in mineral soil. However, several fungi in the roots of either P. abies or Larix sp. showed a certain host specificity.

We investigated the diversity and occurrence of wood wasps in Lithuania and determined communities of associated fungi. Trapping of wood wasps resulted in three different species, including Sirex juvencus, Urocerus gigas, and Tremex fuscicornis. Fungal culturing from adult females of T. fuscicornis mainly resulted in fungi from the genera Penicillium and Trichoderma. High-throughput sequencing of ITS2 rDNA resulted in 59,797 high-quality fungal sequences, representing 127 fungal OTUs. There were 93 fungal OTUs detected in U. gigas, 66 in S. juvencus, and 10 in T. fuscicornis. The most common fungi were Fusarium sporotrichioides (63.1% of all fungal sequences), Amylostereum chailletii (14.9%), Penicillium crustosum (7.8%), Microascus sp. 2261_4 (5.0%), and Pithoascus ater (2.1%). Among these, only A. chailletii was found in all three insect species with the highest relative abundance in U. gigas (15.2%), followed by S. juvencus (7.7%), and the lowest in T. fuscicornis (0.3%) (p < 0.0003). Correspondence analysis of fungal communities showed a distant placement of different species of wood wasps, indicating that fungal communities in each of these were largely different. In conclusion, the study showed that the economically important tree pathogen A. chailletii was among the most common fungal OTUs associated with siricid wood wasps.

In 2019, the bark beetle Ips amitinus (native to central Europe) was identified in south-western Siberia at a distance exceeding 2500 km east of its previously known easternmost location in the European part of Russia. In Siberia, its invasive populations are characterised by high abundance and harmfulness. Here, I. amitinus accomplishes primary attacks on standing vital trees of Pinus sibirica with a lethal outcome. This invasion has already resulted in massive dieback in stands of pine over a large geographic territory. By, 2021, the invaded area was estimated to cover at least 31,200 km2. The objectives of this study were to investigate fungi associated with/vectored by I. amitinus in its invasive area in south-western Siberia and wood decay fungi that cause root and butt rots to P. sibirica. This led to the following conclusions: (i) DNA analysis of sixty adult beetles of Ips amitinus collected from P. sibirica in south-west Siberia revealed the presence of 143 fungal taxa; (ii) species richness was significantly higher in beetles collected from dead branches than from (more recently infested) dying branches; (iii) fungal communities were >90% dominated by yeasts, among which the most common were Nakazawaea holstii, Kuraishia molischiana, and N. ambrosiae; (iv) entomopathogenic Beauveria bassiana s.l. was the most common fungus isolated from dead/mycosed beetles of I. amitinus, followed by Lophium arboricola and four Ophiostoma spp.; and (v) Heterobasidion parviporum was the most common decay fungus detected, which was causing heart rot in stems of P. sibirica.

Abies concolor was introduced to Europe in the 19th century and commonly planted as an ornamental tree. In 2018, after several very warm and dry vegetation seasons, which has likely caused abiotic stress in the trees, a sudden dieback of A. concolor was observed in the city of Aranđelovac in Serbia. The external symptoms were the rapid discoloration of needles in a larger part of the crown and eventually the dieback of the trees. The aim of this study was to identify the possible agents of damage by analysing the fungal communities associated with symptomatic needles using high-throughput sequencing as no symptoms of damage were seen on the branches or stems. Symptomatic needles were collected from the lower branches of five standing trees. Half of the needles were surface sterilised and the remaining were left unsterilised. DNA was extracted, amplified using ITS2 rDNA as a marker and sequenced. Quality filtering showed the presence of 6 191 high quality reads, which, following a clustering analysis, was found to represent 79 non-singletons fungal taxa. The most common fungi in both surface sterilised and unsterilised needles were the pathogens Cytospora friesii (59.7%) and Sydowia polyspora (20.6%). The results demonstrated that C. friesii and S. polyspora can cause a rapid decline of A. concolor trees subjected to abiotic stress.

The aim of this study was to obtain a better understanding on short-distance basidiospore dispersal of the wood-decay fungus Fistulina hepatica, thereby providing valuable knowledge for the conservation management of this protected species. Specifically, the study was expected to reveal site-specific patterns of basidiospore release and spread in oak stands during one fruiting season under north European conditions. The trapping of fungal spores was carried out between August and October 2022 using passive spore traps placed in three oak stands (>200-year-old) in central Lithuania. The average daily temperature was recorded throughout the period of spore trapping. Collected samples were analyzed using high-throughput sequencing of fungal ITS2 rDNA. The results showed that the relative abundance of F. hepatica reads was influenced by the time of fruitbody maturation, but not by the average daily temperature. Although there was a certain variation among different study sites, the results showed that a great majority of F. hepatica spores were deposited within 50 m from the fruitbody, showing that the fungus to a large extent is dependent on local habitats for colonization.

We investigated fungal communities in oak wounds to determine how fungal species richness and community composition changes depending on the age of wounds. The sampling of wood cores was carried out from 10-, 20-, 30-, 40-, and 50-year-old wounds. The fungal community was analyzed using high-throughput sequencing of ITS2 rDNA. Sequence analysis showed the presence of 534 fungal OTUs, which were 83.4% Ascomycota, 16.3% Basidiomycota, and 0.3% Mucoromycota. The fungal OTU richness changed over time: it increased as compared between 10- and 20-year-old wounds, remained similar in 20- to 40-year-old wounds, and decreased in 50-year-old wounds. The fungal community composition also changed over time with the largest differences detected between 10-year-old and older wounds (p < 0.001). The most common representatives of Basidiomycota were Laetiporus sulphureus (34.7%), Mycena galericulata (17.0%), and Cylindrobasidium evolvens (6.5%), and the most common of Ascomycota were Aposphaeria corallinolutea (13.6%), Sclerostagonospora cycadis (7.6%), and Cadophora malorum (5.8%). In conclusion, oak wounds of different ages were colonized by a high diversity of fungi including oak-associated species. Fungal communities in oak wounds underwent qualitative and quantitative changes over time, which led to the gradual shift from fungal generalists in young wounds to oak specialists in older wounds.

We assessed the diversity and composition of fungal communities in different functional tissues and the rhizosphere soil of Pinus sylvestris and Picea abies stands along the latitudinal gradient of these tree species distributions in Europe to model possible changes in fungal communities imposed by climate change. For each tree species, living needles, shoots, roots, and the rhizosphere soil were sampled and subjected to high-throughput sequencing. Results showed that the latitude and the host tree species had a limited effect on the diversity and composition of fungal communities, which were largely explained by the environmental variables of each site and the substrate they colonize. The mean annual temperature and mean annual precipitation had a strong effect on root fungal communities, isothermality on needle fungal communities, mean temperature of the warmest quarter and precipitation of the driest month on shoot fungal communities, and precipitation seasonality on soil fungal communities. Fungal communities of both tree species are predicted to shift to habitats with a lower annual temperature amplitude and with increasing precipitation during the driest month, but the suitability of these habitats as compared to the present conditions is predicted to decrease in the future.

Diplodia sapinea is a common forest pathogen on Pinus spp. in a large part of the world. In 2013, disease caused by this pathogen on Scots pine (Pinus sylvestris) trees in Sweden was reported for the first time. In this study, we report the first detection of D. sapinea on diseased seedlings of P. sylvestris from two Swedish forest nurseries. Infected seedlings were collected July-November 2019. Diplodia sapinea was identified by morphological characteristics of fungal structures on plant tissues and from culture grown on Hagem agar media, followed by sequencing of fungal ITS rDNA. The result emphasizes the susceptibility of P. sylvestris seedlings. More research is needed to better understand the risk for disease spreading within forest nurseries and into the forest through infected plant material.

The genus Phytophthora, with 326 species in 12 phylogenetic clades currently known, includes many economically important pathogens of woody plants. Different Phytophthora species often possess a hemibiotrophic or necrotrophic lifestyle, have either a broad or narrow host range, can cause a variety of disease symptoms (root rot, damping-off, bleeding stem cankers, or blight of foliage), and occur in different growing environments (nurseries, urban and agricultural areas, or forests). Here, we summarize the available knowledge on the occurrence, host range, symptoms of damage, and aggressiveness of different Phytophthora species associated with woody plants in Nordic countries with a special emphasis on Sweden. We evaluate the potential risks of Phytophthora species to different woody plants in this geographical area and emphasize the increasing threats associated with continued introduction of invasive Phytophthora species.

The aim of the present work was to study seasonal and site-specific patterns of airborne fungal diversity, focusing on plant pathogens. The sampling of fungal spores was carried out for twelve months, i.e., between September 2017 and August 2018, using passive spore traps that were placed at three different sites in western (Lenkimai), central (Dubrava), and eastern (Labanoras) Lithuania. Samples were collected every 7–10 days, resulting in 146 samples altogether. Following DNA isolation, samples were individually amplified using ITS2 rRNA as a marker and subjected to high-throughput sequencing. Clustering and taxonomic classification of 283,006 high-quality reads showed the presence of 805 non-singleton fungal taxa. The detected fungi were 53.4% Ascomycota, 46.5% Basidiomycota, and 0.1% Mucoromycota. The most common fungal taxon at Labanoras and Lenkimai was Hannaella coprosmae (23.2% and 24.3% of all high-quality fungal sequences, respectively), while at Dubrava it was Cladosporium macrocarpum (16.0%). In different sites, plant pathogenic fungi constituted between 1.6% and 14.6% of all fungal taxa and among these the most common were Protomyces inouyei (4.6%) and Sydowia polyspora (1.9%). The results demonstrated that the diversity of airborne fungi was mainly determined by the surrounding vegetation and climatic factors, while the occurrence of pathogenic fungi was affected by the availability of their hosts.

In Europe, Pinus sylvestris and Picea abies are the most common coniferous tree species used in commercial forestry, which rely on high‐quality reproductive material for successful reforestation. Clear‐cut harvested forest sites are often replanted using tree seedlings, which are produced in forest nurseries using seeds from seed orchards. However, incidences of fungal diseases in seedling production show that a better knowledge of seedborne fungi, including fungal pathogens, is needed to manage diseases in forest nurseries. This study aimed to assess seedborne fungal communities associated with commercial seeds of P. abies, P. sylvestris and Larix sp. seeds originated from geographically separated regions in Sweden, Belarus, Finland and Poland. Fungal communities were obtained first from the seed surface and then from the seed tissue. These were analysed using high‐throughput sequencing of the ITS2 rDNA region. The results showed that fungal diversity and community composition differed between the seed surface and the seed tissue. Picea abies accommodated a higher fungal diversity than P. sylvestris. In addition, a strong host affinity of the fungal community composition on the seed surface and a weaker association in the seed tissue was found. Fungal communities on P. abies and P. sylvestris seed surface differed significantly between geographical regions, whereas no regional differences were found in the seed tissue. The seedborne fungal communities included a high proportion of plant pathogens, among which the most abundant were Sydowia polyspora (13.3%), Phoma herbarum (11.2%) and Sirococcus conigenus (3.8%). In conclusion, the results showed (a) characteristic fungal diversity and community composition between the seed surface and the seed tissue; (b) a host‐specific fungal community composition on the seed surface and in the seed tissue; (c) regional difference in fungal communities on P. abies and P. sylvestris seed surface, thus the movement of seeds between different regions can contribute to the spread of fungal diseases; and (d) the presence of a high incidence of seedborne fungal pathogens which suggest a potential need of preventative or control measures to reduce the occurrence of these fungi on the seed surface. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate fungal communities associated with leaves and roots of healthy-looking and declining U. glabra trees. The study was expected to demonstrate whether and how the diversity and composition of fungal communities change in these functional tissues following the infection by Dutch elm disease-causing fungi. The study sites included six U. glabra sites in Lithuania, where leaves and roots were sampled. DNA was isolated from individual samples, amplified using ITS2 rRNA as a marker, and subjected to high-throughput sequencing. The sequence analysis showed the presence of 32,699 high-quality reads, which following clustering, were found to represent 520 non-singleton fungal taxa. In leaves, the fungal species richness was significantly higher in healthy-looking trees than in diseased ones (p < 0.05). In roots, a similar comparison showed that the difference was insignificant (p > 0.05). The most common fungi in all samples of roots were Trichocladium griseum (32.9%), Penicillium restrictum (21.2%), and Unidentified sp. 5238_7 (12.6%). The most common fungi in all samples of leaves were Trichomerium sp. 5238_8 (12.30%), Aureobasidium pullulans (12.03%), Cladosporium sp. 5238_5 (11.73%), and Vishniacozyma carnescens (9.86%). The results showed that the detected richness of fungal taxa was higher in samples collected from healthy-looking trees than from diseased ones, thereby highlighting the negative impact of the Dutch elm disease on the overall fungal diversity.

This study aimed to assess the changes in chemical properties and fungal communities in the upper mineral soil layer in managed Scots pine (Pinus sylvestris L.) ecosystems. Study sites were located in the three largest P. sylvestris massifs in Lithuania, and six sampling sites, representing different development stages of the P. sylvestris forest ecosystem, were selected in each of them: mature P. sylvestris forest stands; clear-cuts of former P. sylvestris mature stand; and the P. sylvestris plantations aging from the 1st to 4th year. High-throughput sequencing was performed to evaluate the soil fungus community at clear-cuts, early-stage reforested sites, and mature forests in Lithuania. This study has shown that, among other chemical soil parameters, the mean concentrations of mineral nitrogen (N), total phosphorus (P), and P2O5 were slightly higher in the clear-cut sites, and significantly higher in the 1st year plantations compared to the mature forests. The quality filtering after PacBio sequencing showed the presence of 60,898 high-quality fungal sequences, and 1143 fungal operational taxonomic units (OTUs). The most abundant fungal OTU in our study was Archaeorhizomyces sp. 5425_1. In total, 70 mycorrhizal fungal OTUs were found in the soil samples at the studied sites. The most abundant ectomycorrhizal fungus identified was Amanita fulva (Schaeff.) Fr. The highest amount of ectomycorrhizal fungal OTUs was found in the clear-cut sites and in the mature forests. The concentrations of mineral N and P2O5 in the upper mineral soil layer did not significantly affect fungal OTUs diversity. Conversely, a relatively strong correlation was obtained between the number of mycorrhizal fungal OTUs and the concentrations of total N and soil organic carbon (SOC), as well as between the numbers of saprotrophic fungal OTUs and the concentration of magnesium ions (Mg2+).

In Lithuania, the dieback of European ash (Fraxinus excelsior L.), caused by alien ascomycete Hymenoscyphus fraxineus, started in the mid-1990s, resulting in a large-scale decline of F. excelsior and its dominated forest habitats. Nevertheless, the recent inventories show the presence of several hundred hectares of naturally regenerated F. excelsior stands. We used seven naturally regenerated sites and three planted progeny trials of F. excelsior to collect leaves, shoots, roots, and the surrounding soil to study ash-associated fungal communities based on high-throughput sequencing. Results showed that fungal communities associated with F. excelsior in re-emerging stands in post-dieback areas were composed of 1487 fungal taxa. Among these, 60.5% were Ascomycota, 37.5%—Basidiomycota, 1.7%—Zygomycota, and 0.2% were Chytridiomycota. Revealed mycobiota was largely composed of endophytic fungal communities as these were dominated by Cladosporium sp., Fraxinicola fraxini (syn. Venturia fraxini) and Vishniacozyma foliicola. Identified mycobiota also included a range of ash-specific fungal taxa. Hymenoscyphus fraxineus occurred in all stands but was not frequent. Cladosporium sp. showed strongest negative correlation with the presence of H. fraxineus. This ascomycete, given its dominance in leaves, shoots and in the organic soil layer, might be the limiting factor for the infection rate or spread of H. fraxineus. Although fungal communities in asymptomatic and symptomatic samples of F. excelsior differed significantly from each other, the majority of the most frequently found fungal taxa were not host-specific, suggesting that these were negligibly affected by ash dieback. Investigated stands in natural F. excelsior habitats exhibited larger diversity of fungal taxa (especially ash-specific), than progeny trials planted on former grasslands, indicating the importance of natural habitats in F. excelsior restoration programs.

The plant- and soil-associated microbial communities are critical to plant health and their resilience to stressors, such as drought, pathogens, and pest outbreaks. A better understanding of the structure of microbial communities and how they are affected by different environmental factors is needed to predict and manage ecosystem responses to climate change. In this study, we carried out a country-wide analysis of fungal communities associated with Pinus sylvestris growing under different environmental conditions. Needle, shoot, root, mineral, and organic soil samples were collected at 30 sites. By interconnecting the high-throughput sequencing data, environmental variables, and soil chemical properties, we were able to identify key factors that drive the diversity and composition of fungal communities associated with P. sylvestris. The fungal species richness and community composition were also found to be highly dependent on the site and the substrate they colonize. The results demonstrated that different functional tissues and the rhizosphere soil of P. sylvestris are associated with diverse fungal communities, which are driven by a combination of climatic (temperature and precipitation) and edaphic factors (soil pH), and stand characteristics.

Fifty-nine fungal taxa, isolated from re-emerging Fraxinus excelsior sites in Lithuania, were in vitro tested against three strains of Hymenoscyphus fraxineus on agar media to establish their biocontrol properties. All tested fungi were isolated from leaves and shoots of relatively healthy Fraxinus excelsior trees (H. fraxineus growth by tested fungal taxa ranged between 16–87%. Occasionally isolated fungal taxa such as Neonectria coccinea, Nothophorma quercina, and Phaeosphaeria caricis were among the most effective fungi inhibiting the growth of H. fraxineus cultures. Among the more commonly isolated fungal taxa, Cladosporium sp., Fusarium sp., Malassezia sp., and Aureobasidium pullulans showed a strong growth inhibition of H. fraxineus.

Recent observations suggest that climate change affects the growth conditions and range of tree species distribution in Europe. This may also have a major effect on communities of different organisms associated with these tree species. We aimed to determine whether Larix sp. could provide suitable habitats to insects and lichens associated with P. abies to conserve their biodiversity under climate change. The study sites were 10 Larix sp. and 10 P. abies forest stands in Lithuania. Both living and dead trees were included. Sticky traps, bark sheets, and exit hole methods were used for the assessment of insects. Independent plots on tree stems were established for the assessment of lichens. There were 76 and 67 different insect species on dead and living P. abies, respectively, using sticky traps. Similarly, there were 64 and 68 on dead and living Larix sp., respectively. The overall community of xylophagous insects consisted of nine and eight species, which were detected using the bark sheet and exit hole methods, respectively. The bark area colonized by lichens was 34.3% on dead P. abies and 63.2% on dead Larix sp., and 40.4% on living P. abies and 78.0% on living Larix sp. Taken together, the results demonstrate that native P. abies and introduced Larix sp. support similar diversity of stem-associated insect and lichen species.