LAUSR

Fungal species have numerous important environmental functions. Where these functions occur will depend on how fungi are spatially distributed, but the spatial structures of fungal communities are largely unknown, especially in understudied hyperdiverse tropical tree canopy systems. Here we explore fungal communities in a Costa Rican tropical rainforest canopy, with a focus on local‐scale spatial structure and substrate specificity of fungi. Samples of ~1 cm3 were collected from 135 points along five adjacent tree branches, with intersample distances from 1 to 800 cm, and dissected into four substrates: outer host tree bark, inner bark, dead bryophytes and living bryophytes. We sequenced the ITS2 region to characterize total fungal communities. Fungal community composition and diversity varied among substrate types, even when multiple substrates were in direct contact. Fungi were most diverse in living bryophytes, with 39% of all operational taxonomic units (OTUs) found exclusively in this substrate, and the least diverse in inner bark. Fungal communities had significant positive spatial autocorrelation and distance decay of similarity only at distances less than 1 m. Similarity among samples declined by half in less than 10 cm, and even at these short distances, similarities were low with few OTUs shared among samples. These results indicate that community turnover is high and occurs at very small spatial scales, with any two locations sharing very few fungi in common. High heterogeneity of fungal communities in space and among substrates may have implications for the distributions, population dynamics and diversity of other tree canopy organisms, including epiphytic plants.