Ash dieback, caused by the pathogen Hymenoscyphus pseudoalbidus, is an emerging lethal disease of Fraxinus excelsior in large parts of Europe. To develop a method for the early detection of H. pseudoalbidus, we designed primers for 46 microsatellites (simple sequence repeats, SSRs) of the pathogen. Seven pairs of primers (SSR38, SSR58, SSR114, SSR198, SSR206, SSR211 and SSR212) were found to bind only to the genome of H. pseudoalbidus, but not to the genome of H. albidus or to 52 different fungal endophytes isolated from F. excelsior and F. angustifolia. Using these seven primer pairs, H. pseudoalbidus was identified in fruiting bodies and different types of ash tissues including dead leaves, dead petioles and discoloured or non-discoloured wood. Along one twig, H. pseudoalbidus was detected at different levels of intensity, which depended on the distance from symptomatic tissue. The detection limit was 0.9-1.8 pg of genomic DNA per PCR. Of 50 analysed commercially available seedlings, six were infected with H. pseudoalbidus. Two SSR loci (SSR198 and SSR211) showed fragment length polymorphism. Our results showed that the new primers not only provide an easy and inexpensive means of detecting H. pseudoalbidus in ash tissues, but can also provide information on the genetic heterogeneity of the species.

Abstract:The present paper analyses the community structure of ectomycorrhiza (ECM) and arbuscular mycorrhiza (AM) fungi associated with seven different poplar clone types growing in a patch system on soil from four different former land use types, originating from spruce forest, poplar stand, grassland and cornfield. We determined the extent to which ECM and AM play a role on the studied factors (genotype, former land use type and host growth). The diversity of ECM and AM fungal communities was estimated by morphological and molecular analyses of the 18S and ITS of the rDNA genes. Fifteen ECM fungal taxa and four AM groups were distinguished in the roots of the poplars grown for 18 months on soil originating from the respective land use types. The poplar clones showed significantly different rates of shoot length and AM colonization, especially concerning the occurrence of Glomus intraradices and Scutellospora sp. Populus deltoides had significantly higher Scutellospora sp. abundance. Although ECM abundance and OPEN ACCESSForests 2014, 5 2981 diversity was high, no significant differences between the different land use types was found. However, some ECM fungi like Paxillus involutus, Laccaria proxima and Laccaria tortilis showed significant preferences for specific land use types. Our findings suggest that both factors, former land use type and poplar genotype, are important determinants of mycorrhizal colonization of the host plants.

Ectomycorrhiza (ECM) communities can be described on a species level or on a larger scale at an ecosystem level. Here we show that the species level approach of successional processes in ECM communities is not appropriate for understanding the diversity patterns of ECM communities at contaminated sites. An ecosystem based approach improves predictability since different biotic and abiotic factors are included. However, it still does not take into account the hierarchical structure of the ecosystem. We suggest that diversity patterns of ECMs communities in forests can best be investigated at three levels. This hypothetical approach for investigation can be tested at sites of secondary succession in areas contaminated with metals. Once the diversity patterns are appropriately described by a hierarchical ecosystem approach, to the species level is used to explain these patterns by populational and ecotoxicological mechanisms.

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