Genetic Variability of Hemiscorpius lepturus in Khuzestan Province, Iran, Using ISSR-PCR and Mitochondrial Cytochrome C Oxidase Subunit I Gene Sequencing

Background: Scorpion stings pose a serious public health concern, particularly in Khuzestan Province, Iran. The genus Hemiscorpius scorpions are a major cause of scorpion stings and related deaths, mostly in children. Objectives: This study aimed to study the potential intraspecific variability of Hemiscorpius lepturus in Khuzestan. Methods: We used inter-simple sequence repeat anchored-polymerase chain reaction (ISSR-PCR) and cytochrome c oxidase subunit ۱ (COI) to study potential intraspecies variability of this scorpion. Twenty-two specimens of H. lepturus scorpions were collected from ۵ geographically distinct regions of Khuzestan. Genomic DNA was extracted using the phenol/chloroform method. For phylogenetic analysis, target gene fragments were amplified using ISSR-PCR. By agarose gel electrophoresis of the PCR products, bands produced in each specimen were categorized using a zero and one system, and a dendrogram was drawn using the UPGMA algorithm.Results: ISSR-PCR generated ۵ bands ranging from ۰.۹ to ۲.۵ kb. The results showed that the specimen Hl۴Ch was clearly different within its group. To validate these findings, a ۶۳۷-nucleotide fragment of the COI gene was amplified and sequenced from ۵ genetically variable specimens. Out of ۵ sequences, Hl۱Ba and Hl۶Be have a relatively close relationship (۵۷%) with the H. lepturus reference sequence (KU۳۴۱۹۸۷). However, Hl۴Ch was placed with a relatively high distance (۷۲%) from the rest of them, next to the other reference sequence Hemiscorpius sp. (OP۴۳۳۷۶۲.۱). Conclusion: Although the scorpions of each region were mostly placed together in the phylogeny tree, no major genetic diversity related to regional differences was observed in the province. Based on the genetic distance of Hl۴Ch from other sequences (۱۲.۵%), it is definitely an intraspecies variation.

کلیدواژه ها:

dendrogram ، Inter simple sequence repeat (ISSR) ، Khuzestan ، phylogeny ، Scorpion