Response of juveniles of seven forest tree species and their populations to different combinations of simulated climate change-related stressors: spring-frost, heat, drought, increased UV radiation and ozone concentration under elevated CO2 level.

The study aimed to assess response of juvenile progeny of seven forest tree species, Pinus sylvestris, Picea abies, Betula pendula, Alnus glutinosa, Populus tremula, Quercus robur and Fraxinus excelsior, and their populations to different combinations of climate change-related multiple stressors, simulated in a phytotron under elevated CO2 concentration: (1) heat + elevated humidity (HW); (2) heat + frost + drought (HFD); (3) heat + elevated humidity + increased UV-B radiation doses + elevated ozone concentration (HWUO); and (4) heat + frost + drought + increased UV-B radiation doses + elevated ozone concentration (HFDUO). Effects of the complex treatments, species and species-by-treatment interaction were highly significant in most of the growth, physiological and biochemical traits studied, indicating general and species-specific responses to the applied treatments. For deciduous trees, height increment was much higher under HW treatment than in ambient conditions (control) indicating a positive effect of elevated temperature and better water and CO2 availability. HFD treatment caused reduction of height increment in comparison to HW treatment in most species except for Q. robur and F. excelsior which benefited from lower humidity. Treatments HWUO and HFDUO have caused substantial damages to leaves in fast growing deciduous P. tremula, A. glutinosa and B. pendula, and resulted in their lower height increment than in HW treatment, although it was the same or even higher than that in the control. Rates of photosynthesis in most of the tree species were greatest in HFD treatment. A lower photosynthetic rate (compared to control) was observed in B. pendula, P. tremula and F. excelsior in HW treatment, and in most species-in HWUO treatment. Compared to control, intrinsic water use efficiency in all treatments was significantly lower in P. tremula, A. glutinosa and F. excelsior and higher in conifers P. sylvestris and P. abies. Significant population-by-treatment interactions found for most traits showed variation in response of populations, implying that this reflects adaptive potential of each tree species. The observed responses may not always be considered as adaptive as deteriorating growth of some populations or species may lead to loss of their competitiveness thus compromising regeneration and natural successions.

Population genetic analysis of a parasitic mycovirus to infer the invasion history of its fungal host.

Hymenoscyphus fraxineus mitovirus 1 (HfMV1) occurs in the fungus Hymenoscyphus fraxineus, an introduced plant pathogen responsible for the devastating ash dieback epidemic in Europe. Here, we explored the prevalence and genetic structure of HfMV1 to elucidate the invasion history of both the virus and the fungal host. A total of 1298 H. fraxineus isolates (181 from Japan and 1117 from Europe) were screened for the presence of this RNA virus and 301 virus-positive isolates subjected to partial sequence analysis of the viral RNA polymerase gene. Our results indicate a high mean prevalence (78.7%) of HfMV1 across European H. fraxineus isolates, which is supported by the observed high transmission rate (average 83.8%) of the mitovirus into sexual spores of its host. In accordance with an expected founder effect in the introduced population in Europe, only 1.1% of the Japanese isolates were tested virus positive. In Europe, HfMV1 shows low nucleotide diversity but a high number of haplotypes, which seem to be subject to strong purifying selection. Phylogenetic and clustering analysis detected two genetically distinct HfMV1 groups, both present throughout Europe. This pattern supports the hypothesis that only two (mitovirus-carrying) H. fraxineus individuals were introduced into Europe as previously suggested from the bi-allelic nature of the fungus. Moreover, our data points to reciprocal mating events between the two introduced individuals, which presumably initiated the ash dieback epidemic in Europe.

Genetic Structure in the Northern Range Margins of Common Ash, Fraxinus excelsior L.

During post glacial colonization, loss of genetic diversity due to leading edge effects may be attenuated in forest trees because of their prolonged juvenile phase, allowing many migrants to reach the colonizing front before populations become reproductive. The northern range margins of temperate tree taxa in Europe are particularly suitable to study the genetic processes that follow colonization because they have been little affected by northern refugia. Here we examined how post glacial range dynamics have shaped the genetic structure of common ash (Fraxinus excelsior L.) in its northern range compared to its central range in Europe. We used four chloroplast and six nuclear microsatellites to screen 42 populations (1099 trees), half of which corresponded to newly sampled populations in the northern range and half of which represented reference populations from the central range obtained from previously studies. We found that northern range populations of common ash have the same chloroplast haplotypes as south-eastern European populations, suggesting that colonization of the northern range took place along a single migration route, a result confirmed by the structure at the nuclear microsatellites. Along this route, diversity strongly decreased only in the northern range, concomitantly with increasing population differentiation and complex population substructures, a pattern consistent with a leading edge colonization model. Our study highlights that while diversity is maintained in the central range of common ash due to broad colonizing fronts and high levels of gene flow, it profoundly decreases in the northern range, where colonization was unidirectional and probably involved repeated founder events and population fluctuations. Currently, common ash is threatened by ash dieback, and our results on northern populations will be valuable for developing gene conservation strategies.

Genetic variation and relationships in Laetiporus sulphureus s. lat., as determined by ITS rDNA sequences and in vitro growth rate.

The aim of this study was to characterise the genetic variation and molecular relationships of the brown rot polypore, Laetiporus sulphureus s. lat., from Europe, South America, Africa, and Asia, using ITS sequences of the nu-rDNA and by comparing the growth rate in vitro. In a NJ analysis of the sequences of 130 individuals of L. sulphureus s. lat., eight distinct clusters emerged, supported by BS values of 70-100%. Within each cluster, the ITS rDNA sequence variation was below 3%. The sequences were also analysed together with Laetiporus sequences available from GenBank. Results demonstrated the possible presence of L. huroniensis in Europe (invalidly named L. montanus) and L. gilbertsonii in South America, and provided more information on the Pan-American and European distribution of one of the clades, currently known in North America as L. sulphureus. L. conifericola formed a separate distinct clade. Moreover, the analysis revealed two unknown Laetiporus taxa in Korea, one in South Africa, and one in Europe. As L. sulphureus is described from Europe (France), and we show that more than one taxon exist here, it is presently not possible to define L. sulphureus s. str. Certain biological differences between some of the clades (in vitro growth rates, chemical composition, and pigmentation) were demonstrated and discussed.

Airborne fungal colonisation of coarse woody debris in North Temperate Picea abies forest: impact of season and local spatial scale.

Coarse woody debris is important for mycodiversity in forest ecosystems, but its availability in managed stands is reduced. Leaving dead wood during felling is suggested as an option to sustain and restore the diversity. However, little is known what fungi would colonise freshly cut wood left on managed sites, and how the colonisation process is influenced by ecological factors. During summer and autumn, 120 freshly cut Picea abies stem sections over 8 cm in diameter were placed upright in mapped locations over two discrete plots separated by 100 m in a north-temperate forest. After seven weeks the sections were collected, and isolation and identification of fungi was done from their upper surfaces. In all 943 fungal strains were isolated, representing 97 species. Species richness in the summer survey was 42.5% higher than during the autumn survey. Low species similarity characterized the different seasons (Sorensen indices: S(S) = 0.36 and S(N) = 0.34) and for 21 species (22%) observation frequency was significantly affected by season. As a result, community structures in summer and autumn differed notably (z-test; P < 0.001). Species richness between the two plots differed by less than 10%, but there were 64 species (66%) found only in one of them, thus qualitative similarity was low (S(S) = 0.49). Quantitative similarity was higher (S(N) = 0.63), indicating that the dominant species colonised wood to a similar extent in both areas. Fungal community structure differed significantly among the two plots (z-test; P < 0.001). Our data showed that freshly cut CWD contributed to mycodiversity in managed north-temperate forest, providing habitats for numerous individuals from over 100 species. The fungal community within a single stand differed markedly both across small distances and over the seasons. In order to sustain and enhance mycodiversity in managed stands. coarse wood should always be left during harvesting. This study also demonstrates the importance of molecular identification and ITS sequence databases for exploring fungal diversity in natural communities.

Clonality in the postfire root rot ascomycete Rhizina undulata.

The aim of the present work was to investigate the potential for territorial and dispersive clonality in natural populations of the postfire root rot ascomycete Rhizina undulata. Population studies based on vegetative compatibility tests were done with strains isolated from individual sporocarps at five burned sites in three different localities (separated by 20-40 km) in the Curronian Spit of western Lithuania. Among a total of 103 strains, the tests identified 14 distinct vegetative compatibility groups (VCGs) of R. undulata, 13 of which were represented by 2-48 strains and three were encountered at 2-4 different sites. Occurrence on spatially separated sites of the same VCG of the fungus indicated a presence of dispersive clonality in R. undulata populations. On a local scale clusters of vegetative compatible sporocarps usually occupied discrete territories, implying territorial clonality. The two largest VCGs covered areas up to 7 and 3 m across. The results show that both dispersive and territorial clones are characteristics of natural populations of the fungus.

Ecology and molecular characterization of dark septate fungi from roots, living stems, coarse and fine woody debris.

The aim of the present work was to determine the identity and molecular relationships between 127 strains of dark septate (DS) fungi isolated from healthy root tips, decayed coarse roots, live healthy-looking stems, coarse (stumps, snags and logs) and fine (tree branches and tops) woody debris in temperate-boreal forests in Sweden and Lithuania. Sequence analysis of ITS rDNA was used to identify the fungi. In a neighbour-joining similarity tree, all sequences were grouped into five distinct clusters. Within each of these, ITS rDNA sequence variation consisted of 2-18 nucleotides, corresponding to 1-3% of their total length. The four least variable clusters were supported with high bootstrap values of 86-100%. Comparisons with the sequences in the GenBank database showed that all our strains had a 95-100% homology with identified Phialocephala species, and they were thus assigned to this genus. The representatives of two clusters were identified, as P. fortinii and P. dimorphospora. The representatives of three remaining clusters were defined as Phialocephala sp. 35, Phialocephala sp. 6 and Phialocephala sp. 18. Within each of these clusters, ITS rDNA sequence uniformity was higher than that observed within P. fortinii and P. dimorphospora. Consequently, their clusters were most discrete, supported with bootstrap values of 100%. Genetic variation in the five distinguished Phialocephala species and their possible ecological roles are discussed. Phialocephala sp. 6 was confined to healthy root tips of conifers. P. dimorphospora was only associated with dead woody tissue of P. abies. P. fortinii, Phialocephala sp. 18 and sp. 35 were isolated from both dead and living conifers and Betula pendula. In conclusion, the present study revealed the ability of fungi from the genus Phialocephala to colonise and persist in live and dead trees under strikingly different ecological conditions.