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Phylogenetic analysis of flatfish species (Teleostei, Pleuronectiformes) based on cytochrome oxidase 1 (Co-1) and cytochrome b (Cyt-b) genes Sharina S.N., Kartavtsev Y.P. INTRODUCTION The phylogenetic relationships within the order are still problematic Fish mitochondrial DNA is now widely used with phylogenetic purposes Most popular in phylogenetics are sequences of cytochrome b (Cyt-b) and cytochrome oxidase 1 (Cо-1) genes, which used for taxa comparison at the speciesfamily level. Aim: Molecular phylogenetic investigation of relationships among order Pleuronectiformes To make reconstruction the phylogenetic relationships based on Co-1 and Cyt-b gene sequencing among some flatfish species were made. To confirm or disprove the monophyly of the flatfish family Pleuronectidae and genus Pleuronectes several tree types were built. Materials and methods Samples of the collection Kievka Bay Vostok Bay (Kartavtsev et al., 2007) Flatfish-cytb-F – ATGGCCAACCTCCGTAAATCCCACCCCCTTC Flatfish-cytb-R – CTGGGGCTCTGGACGCTGAGCTACTAGTGC (~1441 bp) 20 spesies of Pleuronectiformes for Co-1 gene 47 species of Pleuronectiformes for Cyt-b gene 2 species of Gadiformes as outgroup Flatfish-1-F Flatfish-1-R Flatfish-2-F Flatfish-2-R TCTTTAGATTTGCAATCTAACATGTA GTCAGTAGCATTGTAATTCCTGCGGC TGGGCCCATCACATATTTACAGTCGG CAGAGCGGTTATGTGGTTGGCTTGAA (~1500 bp) Rooted consensus (50%) MP-tree showing phylogenetic interrelationships on the basis of Co-1 sequence data. In the nodes bootstrap support (n=1000) for MP and NJ (MP/NJ) are shown. The tree was rooted with the sequences of two out-group species: Gadiformes. Rooted consensus (50%) BA-tree showing phylogenetic interrelationships on the basis of Cyt-b sequence data. In the nodes bootstrap support (n=1000) for NJ and MP and repetition frequencies for n=106 simulated generations for BA (BA/MP/NJ) are shown. The tree was built based on the TrN+I+G model and was rooted with the sequences of two out-group species: Gadiformes. Hippoglossinae Hippoglossoidinae Pleuronectinae Conclusions: Family Pleuronectidae with high confidence may be considered as monophyletic. According the last systematical revision, we have sampled in the genus Pleuronectes only one species P. pinnifasciatus. Other species, placed in different genera Pseudopleuronectes, Liopsetta, and Lepidopsetta. The diagnostics of species (barcoding) based on Co-1 and Cyt- b gene sequences is highly effective because of low intraspecies and high interspecies variability of this markers. THANKS FOR ATTENTION!