Abstract
Western European coastal sand dunes are highly important for nature conservation. Communities of the creeping willow (Salix repens) represent one of the most characteristic and diverse vegetation types in the dunes. We report here the results of the first kingdom-wide fungal diversity assessment in S. repens coastal dune vegetation. We carried out massively parallel pyrosequencing of ITS rDNA from soil samples taken at ten sites in an extended area of joined nature reserves located along the North Sea coast of the Netherlands, representing habitats with varying soil pH and moisture levels. Fungal communities in Salix repens beds are highly diverse and we detected 1211 non-singleton fungal 97% sequence similarity OTUs after analyzing 688,434 ITS2 rDNA sequences. Our comparison along a north-south transect indicated strong correlation between soil pH and fungal community composition. The total fungal richness and the number OTUs of most fungal taxonomic groups negatively correlated with higher soil pH, with some exceptions. With regard to ecological groups, dark-septate endophytic fungi were more diverse in acidic soils, ectomycorrhizal fungi were represented by more OTUs in calcareous sites, while detected arbuscular mycorrhizal genera fungi showed opposing trends regarding pH. Furthermore, we detected numerous red listed species in our samples often from previously unknown locations, indicating that some of the fungal species currently considered rare may be more abundant in Dutch S. repens communities than previously thought.
Highlights
Fungi represent one of the largest groups of living organisms
Ascomycota was the dominant phylum and accounted for 33.18% of the OTUs, followed by Basidiomycota (22.73%) Glomeromycota (5.29%), Mucoromycotina, incertae sedis (1.94%), and Chytridiomycota (0.53%), while unidentified fungal OTUs with most similar sequences to other environmental sequences without assignment to phylum accounted for 36.33% (Figure 2a)
In non-metric multidimensional scaling (NMDS) ordinations, soil pH had high Pearson correlation values with the dominant ordination axis and overrode the effect of the geological district (Wadden vs. Renodunal) on community composition in both the NMDS and the multiresponse permutation procedure (MRPP) analyses (Figures 3 and 4). This is in agreement with the findings of Van der Heijden et al [25] that soil pH significantly contributed to the variation explained in ECM morphotype composition of communities associated with S. repens
Summary
Fungi represent one of the largest groups of living organisms. fungi are still poorly understood and appreciated compared to plants and animals and our knowledge of fungal diversity lags far behind [1]. The task of discovering a significant portion of yet unknown species has only recently become possible with the advent of highthroughput DNA sequencing of environmental samples that has enormous potential to further boost data acquisition in biodiversity research [5],[6],[7],[8],[9],[10],[11],[12],[13],[14],[15],[16] This methodology involves identification of multiple species from environmental samples using targeted loci selected with the purpose of identification. These loci are typically the same ones that are used in specimen-based large-scale DNA barcoding efforts (e.g., [17],[18]), providing both appropriate power of resolution for the group of interest and maximum amount of reference sequences available from vouchered specimens
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