Basidiomes of Amanita alboverrucosa, A. ochrophylla, and A. pyramidifera were collected from native mixed sclerophyll forest sites and of A. conicoverrucosa and A. punctata from planted stands of Eucalyptus maculata in New South Wales, Australia. DNA was extracted from stipe tissue and subjected to inter-simple sequence repeat (ISSR) PCR analysis conducted using the primers (GTG)5 and (GACA)4 in order to determine genotype distribution at each site. Two to nine genotypes of one of the species were identified at each field site. Genotypes of A. ochrophylla, A. conicoverrucosa and A. punctata were spread over areas of ca 10–60 m diam, suggesting vegetative spread via large below-ground mycelial genets. In contrast, genotypes of A. alboverrucosa were more spatially restricted, suggesting recent establishment via basidiospores and more limited below-ground vegetative spread. Two groups of A. pyramidifera basidiomes that were separated by ca 600 m were found to be of the same genotype. While this might reflect long distance spread of below-ground mycelium in this taxon, the proximity of the basidiomes to a roadway makes movement of vegetative basidiome tissue via vehicular activity and subsequent establishment equally plausible.