Examining the fine-scale spatial structure of fungal populations can tell us much about how individual species reproduce and disperse throughout natural landscapes. Here we study the fine-scale genetic structure of Tricholoma matsutake, a prized edible and medicinal mushroom, by systematic sampling of mycorrhizas within fairy rings in 50-y-old and old-growth forests in two villages. Using single nucleotide polymorphism DNA markers we show that mycorrhizas in both forest age classes in both villages showed high levels of genotypic diversity, consistent with a reproductive life history predominated by outcrossing via basidiospore dispersal. Both the percentage of polymorphic loci within fairy rings, as well as genotype diversity were higher in old-growth compared to 50-y-old forests. Fifty-year-old forests showed significant spatial autocorrelation between pairs of mycorrhizas up to 42m, and a pattern consistent isolation-by-distance structure. Spatial patterns in old-growth forests were random. Furthermore, AMOVA analysis indicates that 11% of molecular variance in 50-y-old forests is partitioned between villages, whereas no significant variance is partitioned between villages in old-growth forests. We conclude that populations of T. matsutake in 50-y-old forests are the result of a founder effect maintained by local inoculation sources. This pattern attenuates as forests age and accumulate inocula from more distance sources. We speculate on how genetic mosaicism within T. matsutake fairy rings may structure populations within a chronosequence. Finally, we discuss how population spatial dynamics and dispersal strategy in T. matsutake contrast with other ectomycorrhizal species.