High-throughput sequencing of fungal communities across the perennial ice block of Scărișoara Ice Cave

This survey presents the first high-throughput characterisation of fungal distribution based on ITS2 Illumina sequencing of uncultured microbiome from a 1500 years old perennial ice deposit in Scărișoara Ice Cave, Romania. Of the total of 1 751 957 ITS2 sequences, 64% corresponded to 182 fungal operational taxonomic units, showing a low diversity, particularly in older ice strata, and a distinct temporal distribution pattern. Ascomycota was the major phylum in all ice samples, dominating the 400 and 1500 years old ice strata deposited during the cold Little Ice Age (LIA) and Dark Ages Cold Period, while Basidiomycota was mostly present in 900-years old ice formed during the Medieval Warm Period (MWP). Chytridiomycota and Mucoromycota phyla were present in recently formed and 400-years old ice, respectively. Among the 80 identified genera, Cryptococcus victoriae, commonly found in glacial habitats, was identified in all strata. A positive correlation between fungal distribution and ice conductivity, Ca, Na and Sr concentrations was observed across the ice block, with pH values trailing climate variations during LIA and MWP, respectively. Our record highlighted the presence of a complex climate and environmental-driven fungal community in perennial ice strata accumulated during the last 1500 years in Scărișoara Ice Cave.