Evidence of introgression in endemic frogs from the campo rupestre contradicts the reduced hybridization hypothesis

The campo rupestre ecosystem is considered an old, climatically buffered, infertile landscape. As a consequence, long-term isolation is thought to have played an important role in the diversification of its biota. Here, we tested for hybridization between two endemic leaf frogs from the campo rupestre. We used sequence markers and coalescent models to verify haplotype sharing between the species, to test the existence and direction of gene flow, and to reconstruct the spatiotemporal dynamics of gene flow. Additionally, ecological niche modelling (ENM) was used to assess for potential co-occurrence by overlapping the climatic niche of these species since the middle Pleistocene. We found haplotype sharing and/or lack of differentiation in four nuclear fragments, one of them associated with introgression. The coalescent models support introgressive hybridization unidirectionally from Pithecopus megacephalus to P. ayeaye, occurring ∼300 kya. ENM corroborates this scenario, revealing areas of potential environmental niche overlap for the species at about 787 kya. These results contradict the expectation of reduced hybridization, while ENM suggests climatic fluctuation rather than stability for the two species. The reduced hybridization hypothesis needs to be further investigated because our results suggest that it may have unrealistic premises at least for animals.