A study was carried out to clarify the role of the fungus Chalara fraxinea in decline of Fraxinus excelsior , which is observed on a large scale in central and northern Europe with high incidence of tree mortality. The aims of this work were: (i) to check for the presence of C. fraxinea in various tissues of declining F. excelsior by agar culture isolations and by direct analysis of plant tissues using molecular techniques (DNA extraction, ITS-PCR, cloning, ITS sequencing and T-RFLP); (ii) to study fungal communities inhabiting tissues with symptoms; and (iii) to test the pathogenicity of C. fraxinea to F. excelsior . Chalara fraxinea was isolated from 93% of stem cankers, 91% of necrotic leaf stalks, 27-28% of bark wounds and 30% of visually healthy leaf stalks. Molecular analyses of necrotic leaves, leaf stalks and bark revealed the presence of 25 different fungal taxa, 14 of which were detected in all three types of tissue sample. Chalara fraxinea was the second most common species (61% of samples), and only Cryptococcus foliicola occurred more often (70%). All eight of the tested C. fraxinea isolates induced necroses in bark and cambium on each of 86 inoculated trees, and all controls remained healthy. Average length of necroses caused by different C. fraxinea strains varied from 4·2 to 8·9 cm, but the differences were statistically insignificant. Instead, differences in resistance of individual trees to C. fraxinea were observed.

Currently, massive dieback of Fraxinus excelsior is observed in countries of eastern, northern and central Europe, and the reasons for it are unclear. The aims of the present work were (a) to study fungal communities in declining F. excelsior crowns; (b) to clarify role of fungi in the decline. Shoots from symptomatic crowns were collected in four localities in central Sweden, and distributed into the following categories: (a) visually healthy; (b) initial necroses; (c) advanced necroses; (c) dead tops. The most frequently isolated fungi were Gibberella avenacea, Alternaria alternata, Epicoccum nigrum, Botryosphaeria stevensii, Valsa sp., Lewia sp., Aureobasidium pullulans and Phomopsis sp., and these taxa were consistently found in shoots of all four symptomatic categories. Forty-eight taxa of other fungi were isolated, and fungal diversity was not exhausted by the sampling eVort. The same taxa of fungi were dominant in F. excelsior shoots of diVerent symptomatic categories, and moderate to high similarity of fungal communities was observed in shoots despite the symptoms. Forty-four isolates from 24 fungal taxa were used for artiWcial inoculations of 277 1-year-old F. excelsior seedlings in bare root nursery. After 2 years, only four fungi caused symptomatic necroses of bark and cambium: A. alternata, E. nigrum, Chalara fraxinea and Phomopsis sp. The most pathogenic was C. fraxinea, inducing symptoms on 50% of inoculated trees, while three other fungi caused necroses on 3-17% of inoculated trees. Infection biology of C. fraxinea and environmental factors determining susceptibility of F. excelsior to decline deserve future investigations.

Dynamic conservation of forest genetic resources (FGR) means maintaining the genetic diversity of trees within an evolutionary process and allowing generation turnover in the forest. We assessed the network of forests areas managed for the dynamic conservation of FGR (conservation units) across Europe (33 countries). On the basis of information available in the European Information System on FGR (EUFGIS Portal), species distribution maps, and environmental stratification of the continent, we developed ecogeographic indicators, a marginality index, and demographic indicators to assess and monitor forest conservation efforts. The pan-European network has 1967 conservation units, 2737 populations of target trees, and 86 species of target trees. We detected a poor coincidence between FGR conservation and other biodiversity conservation objectives within this network. We identified 2 complementary strategies: a species-oriented strategy in which national conservation networks are specifically designed for key target species and a site-oriented strategy in which multiple-target units include so-called secondary species conserved within a few sites. The network is highly unbalanced in terms of species representation, and 7 key target species are conserved in 60% of the conservation units. We performed specific gap analyses for 11 tree species, including assessment of ecogeographic, demographic, and genetic criteria. For each species, we identified gaps, particularly in the marginal parts of their distribution range, and found multiple redundant conservation units in other areas. The Mediterranean forests and to a lesser extent the boreal forests are underrepresented. Monitoring the conservation efficiency of each unit remains challenging; however, <2% of the conserved populations seem to be at risk of extinction. On the basis of our results, we recommend combining species-oriented and site-oriented strategies.

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Bakys R., Vasaitis R., Skovsgaard J.P. (2013): Patterns and severity of crown dieback in young even-aged stands of european ash (Fraxinus excelsior L.) in relation to stand density, bud flushing phenotype, and season. Plant Protect Sci., 49: 120-126.The extent and temporal pattern of crown damage (attributed to Hymenoscyphus pseudoalbidus) in even-aged stands of Fraxinus excelsior in relation to bud flushing phenotype, stand density, and season was investigated. Data were collected in 2007 in four statistically designed thinning experiments located in 12-15-years old plantations of ash in Denmark. The study included 21 plots of four contrasting, residual stand densities: (1) 1700-5500 trees/ha (unthinned control plots), (2) 1500 trees/ha, (3) 500 trees/ha, and (4) 100-150 trees/ha. Assessments included estimation of flushing phenotype in May, followed by evaluation of severity of crown damage (percentage of crown killed) in June and September. Simultaneously, for each tree, the presence or absence of crown wilt and dead tops were recorded. The seasonal pattern of disease severity (average crown damage) was similar in all stands, and disregarding stand density the extent of tree crown damage increased significantly towards the end of the growing season (P < 0.005). Disease severity was the worst in unthinned plots, but otherwise unrelated to stand density. Late-flushing trees were most severely affected (P < 0.001). The observed patterns of disease severity are probably associated with ecological features of the pathogen that still remain largely unknown.

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Ten juvenile Fraxinus excelsior half-sib families from two Lithuanian populations have been tested in the controlled environment for their response to ash dieback disease caused by Hymenoscyphus fraxineus, detecting changes of genetic variation and heritability, as well as estimating genotype by environment (G×E) interaction and phenotypic plasticity following artificial spring frost and summer drought treatments. In 2014, a batch of 200 four-year-old ash seedlings was used for each treatment and control (no treatment). Health condition, bud flushing phenology and height were assessed for each seedling, and disease incidence and survival ratios were assessed for each family both before (at the beginning of the vegetation season) and after the treatments (at the end of the vegetation season). Disease incidence ratio increased from 0.77-0.80 up to 0.90-0.95. Tree mortality rates during one vegetation season were significantly lower in the frost treatment (21%) than in the drought treatment (25%) or control (31%). None of the tested F. excelsior families were completely resistant to ash dieback, although significant among-family differences in disease incidence and damage rates suggest an additive mode of gene action and thus a quantitative resistance to the disease. Neither disease incidence rates, nor tree health condition scores differed significantly among the applied treatments (including control) indicating in general a negligible effect of the simulated adverse conditions on health status of the ash seedlings. However, G×E interaction was found to be significant (at P > 0.05) for disease incidence, length of necrotic shoots and tree survival, implying that susceptibility of ash families to the dieback disease unequally depends on environmental conditions, and indicating a presence of genetic variation in plasticity and reaction norms of the tested families across environments (treatments). Substantially increased coefficients of additive genetic variation and heritability in health condition following both frost and drought treatments and compared to control showed that simulated stress conditions may noticeably contribute to expression of differences among the tested F. excelsior families in their resistance traits, thus enabling a better evaluation of performance of different families, an effective family selection for resistance, and achievement of a marked genetic gain.

Ash dieback (ADB) caused by the pathogen Hymenoscyphus fraxineus is the cause of massive mortality of Fraxinus spp. in Europe. The aim of this work was to check for the presence of the molecular marker for ADB tolerance in mapped healthy‐looking F. excelsior trees, and to compare its occurrence in trees exhibiting severe ADB symptoms. Monitoring of 135 healthy‐looking F. excelsior on the island of Gotland, Sweden, showed that after 3–4 years 99.3% of these trees had 0%–10% crown damage, thus remaining in a similar health condition as when first mapped. After 5–6 years, 94.7% of these trees had 0%–10% crown damage. Molecular analysis of leaf tissues from 40 of those showed the presence of the molecular marker in 34 (85.0%) trees, while it was absent in 6 (15.0%) trees. Analysis of leaf tissues from 40 severely ADB‐diseased trees showed the presence of the molecular marker in 17 (42.5%) trees, but its absence in 23 (57.5%) trees (p < .0001). The results demonstrated that monitoring of healthy‐looking F. excelsior is a simple and straightforward approach for the selection of presumably ADB‐tolerant ash for future breeding. The cDNA‐based molecular marker revealed moderate capacity on its own to discriminate between presumably ADB‐tolerant and susceptible F. excelsior genotypes.