Abstract: The influence exerted by tree communities, topography, and soil chemistry on the assembly of macrofungal communities remains poorly understood, especially in highly diverse tropical forests. Here, we used a large dataset that combines inventories of macrofungal Basidiomycetes fruiting bodies, tree species composition, and measurements for 16 soil physicochemical parameters, collected in 34 plots located in four sites of lowland rain forests in French Guiana. Plots were established on three different topographical conditions: hilltop, slope, and seasonally flooded soils. We found hyperdiverse Basidiomycetes communities, mainly comprising members of Agaricales and Polyporales. Phosphorus, clay contents, and base saturation in soils strongly varied across plots and shaped the richness and composition of tree communities. The latter composition explained 23% of the variation in the composition of macrofungal communities, probably through high heterogeneity of the litter chemistry and selective effects of biotic interactions. The high local heterogeneity of habitats influenced the distribution of both macrofungi and trees, as a result of diversed local soil hydromorphic conditions associated with contrasting soil chemistry. This first regional study across habitats of French Guiana forests revealed new niches for macrofungi, such as ectomycorrhizal ones, and illustrates how macrofungi inventories are still paramount to can be to understand the processes at work in the tropics. Abstract in Spanish is available with online material. © 2020 The Association for Tropical Biology and Conservation