Assembly of fungal communities remains poorly understood in part because of the daunting range of spatial scales that may be involved in this process. Here, we use individual leaves as a natural sampling unit, comprising spatially distinct habitat and/or resource patches with unique histories and suites of resources. Spatial patterns in fungal beta diversity were tested for consistency with the metacommunity paradigms of species sorting and neutral dynamics. Thirty senesced leaves were collected from the forest floor (O horizon) in replicate upland forest, riparian forest and vernal pool habitats. We quantified spatial distance between leaves, and fungal community composition was assayed by terminal restriction fragment length polymorphism. Significant distance-decay relationships were detected at all but one upland site. This is the first study where changes in fungal community composition were quantified across discrete adjacent habitat patches, providing evidence that fungal distance decay is operational at a scale of centimetres. Although leaves of differing lignin contents were sampled from each site, leaf type was not consistently important in explaining variation in fungal community composition. However, depth of a leaf within the forest floor significantly influenced community composition at five of six sites. Environmental heterogeneity associated with depth could include moisture gradients, relative influence of soil or spore colonization, and impact of forest floor biotic community (i.e. collembola and earthworms). Because the influence of spatial distance and depth on fungal community composition could not be disentangled, both species-sorting and neutral processes may be embedded within the distance-decay relationships that we found.