show Abstracthide AbstractLions were once the most globally widespread mammal species, with distinct populations in Africa, Eurasia, and America. We generated a genomic dataset that included 2 extinct Pleistocene cave lions, 12 lions from historically extinct populations in Africa and the Middle East, and 6 modern lions from Africa and India. Our analyses show the level of population structure and admixture between lion clades, identifying the Pleistocene cave lion as maximally distinct with no evidence of hybridisation with other lion groups. We also confirm longterm divisions between other lion populations, and assess genetic diversity within individual lion samples. Our work clarifies the complex nature of the global lion species-complex and its evolution, and provides conservation data for modern lion regional populations.