Orthochirus sp. (Family: Buthidae) is widespread throughout the world. There are 17 species, 7 of which have been reported in Khuzestan Province of Iran, including the most diverse species related to the scorpion. In the present study, specimens of O. iranus, O. farzanpay, O. zagrosensis, and O. stockwelli were collected from different areas in this province. Phylogenetic analysis was conducted through duplicating and sequencing the cytochrome c oxidase subunit 1 (COXI) and 16SrRNA genes and comparing their genetic diversity. Orthochirus iranus had the highest genetic distance from O. stockwelli, while the lowest genetic distance was observed between O. farzanpay and O. stockwelli. In addition, the highest interspecific divergence was reported between the species of O. iranus and O. zagrosensis, while the lowest interspecific divergence was found between the species of O. farzanpay and O. stockwelli. The results indicated that cytochrome c oxidase (COXI) and 16SrRNA genes have species identification value.
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