This study aimed to analyze spatial patterns of soil ectomycorrhizal fungal (EMF) communities at the local scale in a subtropical pine-oak forest located in the Nearctic-Neotropical transition in central Mexico, to underpin biodiversity conservation strategies in forest fragments of this region. We used a spatially-explicit nested square sampling design with the same sampling representativeness at all scales and replicated three times. We detected 674 EMF OTUs within 19,200m2 and 65 OTUs on average per sample. Seventy percent of OTUs were detected in only 1e4 samples. Average community similarity was below 5%, showed minor change within 14 and 339m distance and increased with the spatial grain used to compare the data. We found a high species-area relationship and beta diversity coefficients for soil fungi indicating that, at the local scale, increasing area by a constant factor of four represented an in- crease in OTU richness by a factor of two.