A Study on the Specialization of Anemone Fishes Using Molecular Genetics Techniques

This research investigated the specialization of anemone fishes and the phylogenetic relationships among 12 species of the genus Amphiprion and between two species of the genus Premnas using the mtDNA D-LOOP sequence. Following DNA extraction from caudal fins of 14 fish samples, a 450 bp fragment of the D-LOOP mitochondrial (mt) DNA was amplified through polymerase chain reaction (PCR) using a pair of specific primer. The amplified products were tested on agarose gel (1%). The obtained sequences were registered in the Genbank. The arrays of nucleotides obtained from the samples were corresponded to those available in Genbank using BLAST tool in NCBI database. The results showed significant correlations with D-loop nucleotide arrangements of common anemone fish (Amphiprion ocellaris) and those of other fish types indicating the accurate selection of the desired fragment of mtDNA. Resultant phylogenetic tree revealed species abundance together with distribution of many species located in the end of the tree probably classified as older species. Accordingly, a region at a longitude between the Philippines and large sea cliffs at latitude between Sumatra and Melanesia was determined to be the possible area of origin for Amphiprioninae, which was identified as the most important center of biodiversity and evolution.