Persian leopards Panthera pardus saxicolor have been extirpated from over 84% of their historic range and are now limited to rugged landscapes of West Asia and the Caucasus. Understanding and maintaining genetic diversity and population connectivity is important for preventing inbreeding and genetic drift, both of which can threaten population viability. All previous analyses of intraspecific genetic variation of West Asian leopards based on the NADH dehydrogenase subunit 5 gene have reported low mitogenomic diversity. In the current study, we sequenced 959 bp of the mtDNA cytochrome b gene to describe the spatial genetic structure of 22 wild Persian leopards across Iran, which hosts most of the subspecies extant range. The findings based on phylogenetic trees and median-joining network indicated that leopards from Iran formed a distinct subclade, i.e., P. p. saxicolor. The AMOVA analysis showed significant differentiation (88.55%) between the subclades of Persian leopards and other Asian leopards. The lowest levels of haplotype (0.247) and nucleotide (0.00078) diversity were estimated in Persian leopards from Iran. Mitochondrial genome sequencing revealed only two closely related haplotypes. There was no evidence for recent sudden demographic expansion scenario in Persian leopards. The low diversity in cytochrome b gene could potentially be brought about by selective pressure on mitochondria to adapt to oxidative stress and higher metabolic rates in cold environments
