Wild boar is the most important large game species in Croatia, with populations subjected to hunting/management programs. There are indications from previous analyses of morphological variation and reproductive performance that population structure of wild boar in Croatia is strongly influenced by the Dinaric mountain range that separates Croatia in two biogeographic regions: Continental and Mediterranean. Of the two main wild boar haplogroups found in Europe, the most widespread is the E1 clade (present from Portugal to Russia), while the E2 clade is found within the Italian peninsula and on Sardinia. Outside Italy, the E2 clade has to date been found only in archaeological samples from the Mediterranean region of Croatia. The aims of this study were to investigate the hypotheses that (i) the Dinaric Mountains act as a natural boundary separating the Continental and Mediterranean wild boar subpopulations in Croatia; and (ii) the mtDNA E2 clade is present in Croatian wild boar. A total of 14 unlinked microsatellite loci were examined in 264 geo-referenced individuals, and the 486-bp fragment of the mtDNA control region sequence was examined in 69 individuals. The results suggest that the region of the Dinaric Mountains. represents a weak boundary between the Continental and Mediterranean subpopulations, and that the predominant factor causing the observed genetic structuring is isolation by distance. Based on these results, it can be proposed that the Croatian wild boar population in the mainland should be managed as a whole. Three variable sites and four haplotypes were obtained from the mtDNA sequences, all of which belonged to the major European clade (E1).