Information about the diversity of tropical microbes, including fungi is relatively scarce. This study addresses the diversity, spatial distribution and host preference of ectomycorrhizal fungi (EcMF) in a neotropical rainforest site in North East Ecuador. DNA sequence analysis of both symbionts revealed relatively low richness of EcMF as compared with the richness of temperate regions that contrasts with high plant (including host) diversity. EcMF community was positively autocorrelated up to 8.5+/-1.0-m distance-roughly corresponding to the canopy and potentially rooting area of host individuals. Coccoloba (Polygonaceae), Guapira and Neea (Nyctaginaceae) differed by their most frequent EcMF. Two-thirds of these EcMF preferred one of the host genera, a feature uncommon in boreal forests. Scattered distribution of hosts probably accounts for the low EcMF richness. This study demonstrates that the diversity of plants and their mycorrhizal fungi is not always related and host preference among EcMF can be substantial outside the temperate zone.