To determine the phylogenetic relatedness of Amanita species, internal transcribed spacers (ITSs) and the 5.8S ribosomal RNA gene were amplified by polymerase chain reaction and then sequenced according to the dideoxy chain termination method using an automatic DNA sequencer. The ITS region provided sufficient variability for phylogenetic analyses within the species. Analyses of the ITS sequence data by distance and parsimony methods revealed that the Amanita species are composed of three distinct groups whose main branch is strongly supported by bootstrap analysis. The Singerian system did not fully correspond to present phylogenetic results based on molecuar data. The amyloidd nature of the spores was still phylogenetically significant and the type of volva as well as the cap color were proved to be additional important characters in Amanita phylogenetics.