F. excelsior is affected by dieback in the major part of its natural geographical range in Europe, which results in economic and ecological losses. The disease is caused by the ascomycetous fungus Hymenoscyphus fraxineus, a pathogen introduced to Europe most probably from East Asia. This paper presents data on fungi identified on F. excelsior trees representing two different stages of ash dieback in Poland. Fungal communities were identified in initial necrotic lesions on living stems and twigs using the classical method of isolation on malt extract agar and morphological and molecular analyses. In dead apical parts of stems and twigs, fungi were identified by microscopic analyses of fruit bodies formed in situ. Seventy-one fungal taxa were found in 720 samples with symptoms of initial or advanced necrosis. The most common fungus detected in initial necrotic lesions in each forest site was Hymenoscyphus fraxineus (59.2 % of analysed samples). Other frequently isolated fungi included Alternaria alternata, Diaporthe eres, Diplodia mutila, Fusarium avenaceum, F. lateritium and Phomopsis spp. Fruit bodies on dead apical parts of stems and twigs were produced mostly by Diaporthe eres, Diplodia mutila, Lophiostoma corticola, Phomopsis spp., Sirodothis sp. and Valsa cypri. Fungal communities from different sites were similar, as shown by high Sørensen similarity index values. Greatest variation in fungal community structure at the initial necrotic stage was realized by D. mutila and F. avenaceum, and at the advanced necrotic stage by D. eres, D. mutila and Phomopsis spp. Data show a close affinity of certain fungi to site, suggesting geographical relatedness. The ecological effects of distribution of the invasive H. fraxineus and of other fungi sporulating on diseased ash trees in Europe are discussed.
A total of 159 colonies of Chalara fraxinea were isolated between 2005 and 2006 from dying trees of European ash (Fraxinus excelsior L.) aged between 3 and 10 years. They derived from five regions of Poland differing by geographic location and climatic conditions. On the basis of 90 RAMS markers, pathogen intra-and inter-population variability, as well as its dependency on geographic distance and climatic conditions in the regions of strain origin, was analysed. The applied measures of intrapopulation genetic variability (genetic distance, Nei's unbiased diversity, Shannon's Information Index and percentage of polymorphic loci) allowed for differentiation of two strain groups: the first deriving from lowlands and the second from uplands and mountainous areas. Strains in lowlands were characterised by smaller number of markers, smaller number of polymorphic loci and smaller intrapopulation genetic variability. Positive and statistically significant correlation was shown between variability of isolates and elevation of regions above sea level. Pair-wise genetic distances between groups of isolates (Nei's unbiased genetic distance) from particular regions were not significantly correlated with the corresponding geographic distances. On the basis of AMOVA, it was shown that 85% of variability was within-region differences and 2% betweenregion differences, whereas differences between lowlands and uplands were 13%. Principal Components Analysis (PCA) for the investigated regions confirmed the results from Nei's genetic distance matrix.
Abstract• Microsatellite markers were used to describe the genetic structure and variability of early, intermediate and late phenological forms of European beech (Fagus sylvatica L.). Two hundred and seventy individuals from three populations located in southern Poland were divided into three forms according to the phenological criterion -bud burst, and analyzed for allelic variation at five highly polymorphic microsatellite loci.• Population differentiation was moderate and differed significantly among phenological forms. • Analysis of Molecular Variance (AMOVA) revealed different genetic structures characteristic of respective phenological forms of beech. The amount of within-population variability increased with the delay of the beginning of vegetation and amounted to 64%, 77% and 90% of total variability, depending on phenological form. A similar trend was found in average pairwise genetic distance between individuals belonging to a given phenological form (11.78, 11.85 and 12.22, from early to late forms).• Our results demonstrate the importance of late spring frosts as a factor influencing the genetic structure of beech, and as a cause of the decrease in genetic variability as well as the increase in population differentiation proportional to the degree of phenological earliness.
A large part of the area in Europe in which Fraxinus excelsior is native is currently affected by ash dieback, a threatening disease caused by the ascomycetous fungus Hymenoscyphus fraxineus. Fungi other than H. fraxineus also occur in large numbers on stems of the dying ash trees. To clarify their possible role in the dieback process, six fungal species common on dying stems and twigs of ash in Poland, i.e. Cytospora pruinosa, Diaporthe eres, Diplodia mutila, Fusarium avenaceum, F. lateritium and F. solani, were tested for pathogenicity using a test based on artificial wound inoculations of 6‐year‐old F. excelsior plants under field conditions, with H. fraxineus included for comparison. There were significant differences in index of pathogenicity among the fungi tested. Hymenoscyphus fraxineus (mean index 5.78) was the most pathogenic. Diplodia mutila (4.23) and C. pruinosa (4.02) were significantly less pathogenic than H. fraxineus, but significantly more than the other fungi. Diaporthe eres (2.43), F. avenaceum (1.92), F. solani (1.86) and F. lateritium (1.08) were the least pathogenic (P < 0.0001). The extent of disease symptoms caused by F. solani and F. lateritium was statistically similar to the control (P = 0.05). All tested fungi were successfully reisolated from inoculated stems. The contribution of the results to understanding the possible role of these fungi in the ash dieback process in F. excelsior, particularly in trees weakened after primary infection by H. fraxineus, is discussed.
The distribution of genetic diversity in a local population of Dothistroma septosporum was determined on a microspatial geographical scale. The study was conducted in a seed orchard of Pinus nigra divided into four plots, each of 0.84 ha in area. Ninety-two isolates were obtained from needles with red band needle blight symptoms. Molecular variance, Mantel test and autocorrelation spatial analyses were performed with the use of random amplified microsatellites markers to test the null hypothesis of a random distribution of genotypes. The groups of isolates from individual plots demonstrated small differences as regards intrapopulation variation. The mean contribution of polymorphic loci for isolate groups was almost 77%, genetic diversity 0.270, while the Shannon coefficient was 0.389. The Mantel test demonstrated a significant, positive correlation between Huff's genetic distance and geographical distance. Significant spatial genetic structure was detected with autocorrelation coefficients being significant in the first two distance classes up to about 8-12 m.
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