Ash decline induced by Hymenoscyphus pseudoalbidus is an emerging disease that severely affects Fraxinus excelsior stands in Europe. There has been an invasive spread of the disease from east to west in Europe over the last decade. Wood discoloration on infected trunks has been reported, but few data are available on the involvement of H. pseudoalbidus in such symptoms. Transport and trade of ash logs could introduce the pathogen into disease-free areas and therefore accelerate its dissemination. The aim of this study was to assess the prevalence and severity of H. pseudoalbidus in ash logs in infested areas located in the northeast of France and to clarify the role of secondary pathogens in ash decline. The results showed that prevalence of H. pseudoalbidus on collar lesions was high in the study area. The pathogen was able to produce conidia from infected wood. Thus, export of ash logs could represent a potential risk for spreading the disease. Involvement of Armillaria spp. in the decline process was confirmed, while no Phytophthora-induced collar lesions were found. Studying both disease prevalence and the age of callus tissues surrounding collar lesions in 60 ash stands enabled the origin of the disease in the study area to be determined.

Decline induced by Chalara fraxinea is an emerging disease that severely affects ash stands in Europe. The disease appears to have an invasive spread from East to West of Europe in the last decade. The teleomorphic stage, Hymenoscyphus pseudoalbidus, that occurs as apothecia on ash rachis in the litter was recently described. The origin of ash decline remains unclear as a cryptic species, H. albidus, a long-established fungus in Europe, could be present in the same niche, and as in Switzerland, H. pseudoalbidus was shown to have been present long before the recent epidemic outbreak. In France, the emerging disease is very recent and clearly restrained to Northeastern France. We thus collected isolates from infected hosts and from apothecia/ash rachis both inside and outside the infected area in France in order to compare them on the basis of pathogenicity towards ash seedlings and sequences of the ITS regions and of three single-copy genes. We showed that two population types exhibiting about 2% base pair polymorphism in the sequences analysed were present in Northern France. The first type, corresponding to H. pseudoalbidus, was present on rachis and infected hosts only in Northeastern France and showed strong pathogenicity towards ash seedlings in inoculation tests. By contrast, the second type, which corresponds to H. albidus, was present throughout Northern France and showed no pathogenicity towards ash seedlings. Our study confirms the results of Queloz et al. (2010) who presented molecular evidences for the existence of two cryptic species, H. albidus and H. pseudoalbidus. The results strongly suggest that Chalara fraxinea/ H. pseudoalbidus is a recent invader in France.

Hymenoscyphus fraxineus is an invasive fungus in Europe and causes a severe decline affecting ash, which began in the late 1990s. One of the symptoms associated with the disease is lesions in the outer bark of the collar area. However, the aetiology of these basal lesions, and in particular the relative roles of H. fraxineus and Armillaria species, is still controversial; moreover, little is known about the influence of environmental factors on the disease epidemiology. This study therefore surveyed 42 plots located in northeastern France, in an area affected by ash decline since 2008, in order to determine which environmental factors condition the severity of lesions associated with H. fraxineus on ash collar. The spatial pattern that is a consequence of the invasive spread of the disease was taken into account in the analysis, using a spatial hierarchical Bayesian model fitted by integrated nested laplace approximation (INLA). Results show that while basal lesions are tightly associated with H. fraxineus, their severity is influenced by the Armillaria species present in the plot. Sites with vegetation indicating moist conditions, or more humid topographical positions, were associated with more developed basal lesions.

International audienceThe use of next-generation DNA sequencing methods, which produce massive volumes of data, has transformed fungal molecular ecology. These technologies offer the opportunity to describe microbial communities in depth and to investigate fungal diversity at a large geographical scale. In particular, due to the emergence of studies conducted at a large scale (countries, continents), the definition of a compromise between the optimal and representative description of local diversity (plot scale) and the management of a greater number of sites for these studies is required. This work was performed to explore the local richness and the structure of the ectomycorrhizal (EcM) fungal and oomycete communities in a temperate beech plot using 454 pyrosequencing technology. The internal transcribed spacer 1 (ITS1) region was amplified and sequenced from fine roots and soil sampled from the two upper horizons. Our analyses suggest that soil could be a good substitute for fine roots in studying EcM fungi. The fungal assemblage exhibited a vertical distribution in the soil profile. Only four Pythiaceae were identified, which was insufficient to study the spatial distribution of this group. The fungal and particularly the ectomycorrhizal (EcM) richness was not correlated with any soil variable (pH, C, N or P), but the spatial structure was explained by the C:N ratio and phosphorus content. Our data reveal the importance of maintaining a minimal representative sampling at the plot scale for comparative geographical studies

Ash dieback is caused by an invasive pathogen Hymenoscyphus fraxineus, which emerged in Europe in the 1990s and jeopardizes the management of ash stands. Although the biological cycle of the pathogen is well understood, its dispersal patterns via airborne spores remain poorly described. We investigated the seasonal and spatial patterns of dispersal in France using both a passive spore-trapping method coupled with a real-time PCR assay and reports of ash dieback based on symptom observations. Spores detection varies from year to year with a detection ability of 30-47% depending on meteorological conditions, which affect both production of inoculum and efficiency of the trapping. Nevertheless, our results are consistent and we showed that the sporulation peak occurred from June to August and that spores were detected up to 50-100 km ahead of the disease front, proving the presence of the pathogen before any observation of symptoms. The spore dispersal gradient was steep, most of inoculum remaining within 50 m of infected ashes. Two dispersal kernels were fitted using Bayesian methods to estimate the mean dispersal distance of H. fraxineus from inoculum sources. The estimated mean distances of dispersal, either local or regional scale, were 1.4 km and 2.6 km, respectively, the best fitting kernel being the inverse power-law. This information may help to design disease management strategies.

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Summary Collybia fusipes is a common cause of root rot on oak in the north of France. Collybia fusipes basidiomes can be as frequent on oaks in stands where no decline of the trees occurs compared with stands where the decline is chronic. This might be explained by differences in the amount of roots damaged by the parasite. To test that hypothesis, 430 oak trees, Quercus petraea, Quercus robur and Quercus rubra, located in six forests were selected. Half of them showed C. fusipes basidiomes at the trunk base. The association between presence of basidiomes and decline of affected trees depended on the forest. The level of infection of each tree by C. fusipes, as well as the crown appearance, the tree height : diameter at breast height ratio, age and sapwood width were determined. The presence of C. fusipes basidiomes was always associated with significant root infections. The crowns of the trees deteriorated with increasing level of root infection and the decline was severe only when the root damage was heavy. Although the decline of trees that were heavily damaged by C. fusipes was severe in some of the stands, in others, it was only mild, and so the differences in tree decline between the stands could not be attributed solely to differences in root infection severity. Trees damaged by C. fusipes seemed not to be subjected to more competition than their undamaged neighbour as reflected by a similar tree height: diameter at breast height ratio.

Forest declines are usually complex multifactorial phenomena that involve interactions between different factors. The possible interaction between different types of mycelial pathogens was investigated through artificial inoculation of oak seedlings, involving two root rot basidiomycetes, Collybia fusipes and Armillaria mellea, and two Phytophthora species, P. cinnamomi and P. cambivora. These pathogens were inoculated onto young Quercus robur saplings in greenhouse conditions, either alone or combining a root rot basidiomycete with a Phytophthora species. Three out of the four Phytophthora spp.*root rot basidiomycete combinations tested resulted in significantly greater damage to the oak host than the sum of the damages induced by the individual pathogens. This positive interaction could be significant in oak decline syndrome.