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Aim Polar and alpine ecosystems appear to be particularly sensitive to increasing temperatures and the altered precipitation patterns linked to climate change. However, little is currently known about how these environmental drivers may affect edaphic organisms within these ecosystems. In this study, we examined communities of plant root-associated fungi (RAF) over large biogeographical scales and along climatic gradients in the North Atlantic region in order to gain insights into the potential effects of climate variability on these communities. We also investigated whether selected fungal traits were associated with particular climates. Locations Austria, Scotland, Mainland Norway, Iceland, Jan Mayen and Svalbard. Taxa Root fungi associated with the ectomycorrhizal and herbaceous plant Bistorta vivipara. Methods DNA metabarcoding of the ITS1 region was used to characterize the RAF of 302 whole plant root systems, which were analysed by means of ordination methods and linear modelling. Fungal spore length, width, volume and shape, as well as mycelial exploration type (ET) of ectomycorrhizal (ECM) basidiomycetes were summarized at a community level. Results The RAF communities exhibited strong biogeographical structuring, and both compositional variation as well as fungal species richness correlated with annual temperature and precipitation. In accordance with general island biogeography theory, the least species-rich RAF communities were found on Jan Mayen, a remote and small island in the North Atlantic Ocean. Fungal spores tended to be more elongated with increasing latitude. We also observed a climate effect on which mycelial ET was dominating among the ectomycorrhizal fungi. Main conclusions Both geographical and environmental variables were important for shaping root-associated fungal communities at a North Atlantic scale, including the High Arctic. Fungal OTU richness followed general biogeographical patterns and decreased with decreasing size and/or increasing isolation of the host plant population. The probability of possessing more elongated spores increases with latitude, which may be explained by a selection for greater dispersal capacity among more isolated host plant populations in the Arctic.