Molecular Phylogeny Determined by Mitochondrial Genes 16S rRNA, COX1 and Cytb of Eleven Anemonefish Species

Sequences of mitochondrial 16S rRNA, COX1 and Cytb genes were determined to establish the phylogenetic relationship among 11 species of anemonefish using the neighbor-joining (NJ), maximum-likelihood (ML) and Bayesian methods with the divergence and genetic distances determined as well. The 3 genes had a combined length of 4 388 bp that consisted of 1 028 polymorphic and 616 parsimonious-information sites. The ratio of transition to transversion was 2.12, and the average A+T content was 54.7%, which was higher than 45.3% for C+G in the genes. It was found that (1) the subfamily Amphiprioninae was monophyly, but the genus Amphiprion was not monophyletic, implying Premnas should be reclassified; (2) the phylogenetic trees grouped the percula complex into a clade that formed the ancestral taxon of anemonefish, but the phylogenetic relationship was uncertain in regard to Premnas biaculeatus; and, while the Bayesian tree placed the percula complex and P. biaculeatus in a same clade, NJ and ML tree classified P. biaculeatus in a clade with subgenera Amphiprion, Paramphiprion and Phalerebus; and, (3) the genetic distance between species suggested a possibility of P. biaculeatus and Premnas epigrammata being identical.