We investigate the range-wide population structure and phylogeography of thuriferous juniper (Juniperus thurifera L.), a species with a highly disjunct distribution in the western Mediterranean. We genotyped a total of 327 individuals from 20 populations using amplified fragment length polymorphisms (AFLP). Different analyses such as principal co-ordinate analysis (PCoA), nonmetric multidimensional scaling of F(ST) distances among populations, unweighted pair group method with arithmetic mean (UPGMA), and Bayesian clustering revealed that the Strait of Gibraltar acted as an efficient barrier against gene flow between the Moroccan and European populations for a very long time, and consequently support that the Moroccan populations should be recognised as a distinct subspecies (J. thurifera L. subsp. africana (Maire) Romo and Boratyńsky). The Algerian population was genetically more closely related to the European than to the Moroccan ones, probably due to dispersal events from Europe to Algeria. With respect to the mainland European populations, our data are not conclusive to reject any of the two following hypotheses: (1) the Iberian Peninsula was subdivided into different gene pools, and was the source for the colonisation of the Pyrenees and the Alps; and (2) the pattern we see today is partly the result of immigration into the Iberian Peninsula, e.g. from the Alps. Finally, the Corsican population was closely related genetically to two northern Iberian populations most probably due to relatively recent long-distance dispersal.