Taxon-specific fungal primers reveal unexpectedly high diversity during leaf decomposition in a stream

Abstract

Traditional techniques for studying the fungal community composition in streams favour the detection and identification of aquatic hyphomycetes. Our objective was to use molecular techniques to determine the presence and contributions of other fungal groups. We designed primers specific for the ITS regions in Ascomycota, Basidiomycota, Chytridiomycota, Zygomycota and Oomycota. The primers were used to amplify DNA from linden, maple, and beech leaves, and birch wood submerged in a stream for 4 weeks in summer, autumn, winter and spring. The amplification products were separated by denaturing gradient gel electrophoresis. Ascomycota were present in large phylotype numbers (up to 21) on all substrates and all dates and represented ≥ 75 % of the fungal biomass. Basidiomycota were the second most abundant group in summer and autumn (up to 13 % on wood) and were absent only on linden and maple in spring. There were consistently large numbers of phylotypes from Chytridiomycota and their relative contribution to the microbial community peaked in winter on all substrates. Oomycota were present in summer and abundant only on wood. Zygomycota were present in low numbers and their estimated contribution to fungal biomass was ≤ 1%. Using primers to target individual groups facilitates a more balanced approach to studying fungal diversity in freshwater ecosystems.