Non-native pests, climate change, and their interactions are likely to alter relationships between trees and tree-associated organisms with consequences for forest health. To understand and predict such changes, factors structuring tree-associated communities need to be determined. Here, we analysed the data consisting of records of insects and fungi collected from dormant twigs from 155 tree species at 51 botanical gardens or arboreta in 32 countries. Generalized dissimilarity models revealed similar relative importance of studied climatic, host-related and geographic factors on differences in tree-associated communities. Mean annual temperature, phylogenetic distance between hosts and geographic distance between locations were the major drivers of dissimilarities. The increasing importance of high temperatures on differences in studied communities indicate that climate change could affect tree-associated organisms directly and indirectly through host range shifts. Insect and fungal communities were more similar between closely related vs. distant hosts suggesting that host range shifts may facilitate the emergence of new pests. Moreover, dissimilarities among tree-associated communities increased with geographic distance indicating that human-mediated transport may serve as a pathway of the introductions of new pests. The results of this study highlight the need to limit the establishment of tree pests and increase the resilience of forest ecosystems to changes in climate.

The species diversity of culturable endophytic fungi was studied in the leaves and twigs of symptomatic and asymptomatic Fraxinus excelsior trees. Endophytic mycobiota was dominated by Ascomycota species, with Pleosporales (44.17%) and Diaporthales (23.79%) endophytes being the most frequently observed in the tree samples. The number of endophytic isolates and species richness varied depending on the sampling date (May and October) and tissue location. Of the 54 species identified based on ITS sequences, 14 were classified as dominant. The most frequently isolated species were Diaporthe eres, followed by Alternaria alternata, Dothiorella gregaria, and Fraxinicola fraxini. The inhibitory effect of 41 species (75 isolates) of endophytes on the radial growth of a Hymenoscyphus fraxineus isolate was studied under in vitro conditions (dual cultures). The radial growth of H. fraxineus was the most inhibited by four endophytic fungi from twigs (Fusarium lateritium, Didymella aliena, Didymella macrostoma, and Dothiorella gregaria). The inhibitory effect of the four isolates was also studied under in planta conditions. The isolates artificially inoculated into the trunks of ash trees reduced the length of necroses formed by H. fraxineus co-inoculated in the same trunks. This effect depended on the isolate, and the inhibition was most prominent only on trunks inoculated with F. lateritium and D. aliena. Although the total length of necrotic lesions formed by the H. fraxineus infection was shorter in the ash trunks co-inoculated with the endophytes, the difference was not significant.

The health of 34 different Fraxinus taxa in association with the pathogenic fungus Hymenoscyphus fraxineus was assessed in four Slovak arboreta. Averaged across all arboreta, nearly one-quarter (24.9%) of all evaluated trees showed ash dieback symptoms. The damage was most serious on the common ash F. excelsior, a native species. The percentage of dead trees did not exceed 2% for all evaluated trees. Generally, ash trees of all ages were affected, though the intensity of the damage varied among the sites. The identity of H. fraxineus was confirmed by conventional PCR targeting the internal transcribed spacer (ITS) sequences of the nuclear ribosomal DNA, as well as the 18S gene/ITS-2 region of the rDNA operon. In Slovakia, the pathogen has expanded its host range from native species not only to their ornamental cultivars, but also to introduced North American (F. cinerea, F. latifolia, F. pennsylvanica, F. quadrangulata) and Asian (F. bungeana, F. chinensis ssp. rhynchophylla, F. mandshurica) ash species. H. fraxineus was also observed on the previous year’s leaf petioles of the native European species F. ornus, considered a weakly susceptible host. In Slovak arboreta, H. fraxineus was found on 23 Fraxinus taxa; 21 of them represent first records for the country. F. bungeana is recorded as a new host species of H. fraxineus.

In temperate regions of Europe, the large pine weevil, Hylobius abietis, is a major pest of coniferous forests mostly at sites where clear-felling is followed by planting of saplings. Control measures against this pest are based on silvicultural techniques, an application of physical barriers on stems of saplings and insecticide treatments. To avoid the use of insecticides, alternative measures such as biological control have been investigated. The goal of the present study was to obtain local strains of entomopathogenic fungi (Ascomycota, Hypocreales) from natural populations of H. abietis, and to investigate their efficacy against the weevil. A survey on entomopathogenic fungi was undertaken at clear-felled areas of spruce forests in northern Slovakia. Two Beauveria species, B. bassiana and B. pseudobassiana, were identified, and 22 in vitro strains were obtained. Mean prevalence of infected adults was low (2.10% ± 0.67%) and the mycosis was mostly recorded during May and June. Virulence of Beauveria strains against the weevil was tested in laboratory. B. bassiana strain AMEP20 was significantly most virulent (LC₅₀ of 0.65 ± 0.10 × 10⁸ conidia mL⁻¹). Treatment with conidia of AMEP20 strain affected feeding damage by the weevil on bark of Scots pine twigs. Daily bark consumption by B. bassiana-treated weevils was lower than by untreated individuals and decreased with increasing conidia concentration used for the treatment. In the outdoor experiment, AMEP20 strain killed weevils that fed on spruce saplings treated with conidia suspensions. Mortality due to mycosis on weevils exposed to the conidia-treated saplings reached 30.0%–76.5% and 55.0%–88.2% after 32 and 46 days, respectively.