Aim Comparison of Epipedobates bassleri (Myers, 1987), which occurs on high-altitude mountain ridges ('sky peninsulas') in the Andean transition zone and demonstrates high levels of divergence in colouration among populations, and Epipedobates hahneli (Schulte, 1999), which occurs throughout the lowland regions of the Amazon basin and is morphologically conserved, using phylogenetic analysis of mitochondrial sequence data and comparison of colour pattern. Location Central cordilleras of Peru (near Tarapoto, San Martin). Methods DNA was extracted from individuals of E. bassleri from the central cordilleras of Peru, and from individuals of E. hahneli from across Peru. The cytochrome b mitochondrial gene region was amplified and sequenced for individuals of each species, and phylogenetic analysis was carried out using Bayesian inference. Genetic distances among populations and geographic distances of each species were examined and compared using Mantel tests. Parametric bootstrapping was used to test the monophyly of E. bassleri. Results Epipedobates bassleri formed a well-supported monophyletic group and showed higher levels of genetic divergence among populations than was shown among populations of E. hahneli from the same region. Distinct clades representing different geographic regions were recovered for E. hahneli. Levels of divergence among more geographically distant populations of E. hahneli were higher than levels of divergence among E. bassleri populations. We found a significant correlation between genetic divergence and geographic distance as measured along a 1000-m contour line, but not as measured by direct routes (crossing putative biogeographical barriers). Main Conclusions Levels of genetic divergence were higher among populations of morphologically conservative E. hahneli than among populations of morphologically variable E. bassleri, suggesting rapid divergence in colouration among populations of E. bassleri. These patterns support previous arguments concerning the role of the montane transition zone between the high mountains and lowlands in divergence and speciation. High levels of both genetic and phenotypic divergence among populations of E. bassleri indicate that ecological or behavioural factors may be responsible for the high levels of colour variation seen among E. bassleri, but not among E. hahnleli, populations.