Pufferfishes belonging to the genus Tetraodonform one of the largest groups in the family Tetraodonti-dae. This group consists of more than 20 species distributedacross freshwater, as well as brackish and coastal waters, ofAfrica and Southeast Asia. However, their phylogeneticand evolutionary relationships are still ambiguous. In thepresent study, mitochondrial genes encoding 16S rRNAand cytochrome b were sequenced for 17 Tetraodon spe-cies collected from various areas mentioned above