Anemone fishes are a group of 28 species of coral reef fishes belonging to the family, Pomacentridae; subfamily, Amphiprioninae and all have an obligate symbiotic relationship with sea anemones. Two species of these small ornamental fishes have been identified in the Persian Gulf including Amphiprion clarkii and Amphiprion sebae. The phylogenetic relationship between Amphiprion species of the Persian Gulf was studied by collecting 15 samples from three Iranian islands, Larak, Farur and Kish. DNA was extracted from each sample and a part of mtDNA was amplified. Two pairs of primers were designed to amplify a final target of 400 bp by nested-PCR. Each amplican was sequenced, aligned and genetic diversity among samples was investigated by phylogenetic analysis. Results show that there is no significant genetic variation among A. clarkii individuals; however, A. sebae individuals from Larak were different from other fishes of the same species. Most probably this is due to the ability of A. clarkii to be symbiotant with all 10 species of host sea anemones which enables it to spread its own population in the 3 islands. However, A. sebae is observed to be symbiotant only with one host in the sea, therefore, has one option that reduces its distribution.