Molecular Characterization and Phylogeny Analysis Based on Sequences of Cytochrome Oxidase gene From Hemiscorpius lepturus of Iran

Hemiscorpius lepturus is a medically important scorpion found along the Iranian borders, especially near to Khuzestan Province in the south-west of Iran. This is the only non-buthid scorpion which is potentially lethal in southern Iran and is responsible for severe dermonecrotic scorpionism. In this study, DNA fragment of the mitochondrial cytochrome c oxidase subunit I (COXI) gene of H. Lepturus for the molecular phylogenetic analysis was amplified. We amplified  a 624 bp gene fragment of cytochrome C oxidase subunit I (COXI) from H. lepturus collected from Khuzestan, Ahvaz by PCR. After sequencing of the PCR products, the phylogenetic analysis was performed using the neighbor-joining method with 1000 replicates of bootstrapping using the MEGA7 software. The results of phylogenetic analysis revealed four distinct clusters (A1, A2, B and C) belonging to the family of Hemiscorpionidae that were grouped together with bootstrap score between 77-96%. The gene fragment of Hl-Kh formed a cluster relative to the only scorpion of H. Lepturus (Hl) from Izeh, Iran with a good bootstrap score of 96. These two samples isolated from the Khuzestan province of Iran are closely related, as they clustered together as Cluster B. The genetic distances of Hl-Kh among the cluster A1, A2 and C ranged from 16 to19% and the lowest interspecific distance was in cluster B between Hl-Kh and Hl (6%). Hl-Kh isolated from the Khuzestan province of Iran are closely related to Hl, as they showed the greatest interspecific variation observed in Hemisccorpius genus in this study. Although the sample size in Cluster B is not large enough to draw a final conclusion, the percentage of sequence divergence was high enough for interspecific comparisons to provide separation of species.