The Laperrine's olive (Olea europaea subsp. laperrinei) is an emblematic species of the Sahelo-Saharan Mountains. Populations of this tree are locally threatened by extinction due to climatic vicissitudes and human activities, particularly in Niger and Algeria. In order to study the spatial genetic structure and the dynamics of O. e. laperrinei populations, we sampled trees in four isolated mountain ranges (Tassili n'Ajjer and Hoggar (Algeria), Tamgak and Bagzane (Niger)). A total of 237 genets were identified using nuclear microsatellites. Phylogenetic reconstruction based on plastid DNA data supported a maternal origin of O. e. laperrinei populations in South Algeria, where a higher allelic richness was observed. Based on nuclear microsatellite data, two levels of structure were revealed: first, individuals from Niger and Algeria were separated in two distinct groups; second, four less differentiated clusters corresponded to the four studied mountain ranges. These results give support to the fact that desert barriers have greatly limited long distance gene flow. Within populations, pairwise kinship coefficients were significantly correlated to geographical distance for Niger populations but not for Algerian mountains. Historical factors and habitat heterogeneity may explain the differences observed. We conclude that the Hoggar acts as an important genetic reservoir that has to be taken into account in future conservation programmes. Moreover, very isolated endangered populations (for example, Bagzane) displaying evident genetic particularities have to be urgently considered for their endemism.