Roads can influence all modes of evolutionary change—natural selection, mutation, genetic drift, and gene flow. Roads can be strong sources of selection for wildlife through direct collision, pollution, and modification of surrounding habitats. In addition to the immediate challenge posed to persistence, road-induced selection can drive phenotypic and genetic divergence between populations across small spatial scales and after just a handful of generations. Some animal populations may rapidly evolve in response to selection, locally adapting to the many impacts of roads. Yet, other populations might fail to respond to selection, for instance if heritability of existing adaptive variation is low. Furthermore, roads may induce phenotypic plasticity, and generalist species may be more resilient than habitat specialists to the habitat alterations caused by roads. Roads can also increase genetic fragmentation of animal populations, leading to a significant reduction of their adaptive potential by decreasing gene flow and genetic diversity, therefore increasing the risk of inbreeding depression, especially for more isolated populations. Overall, evolutionary perspectives are critically needed in studies of road effects on animal populations.