Abstract Key message J. sabina var. balkanensis, that is of hybrid origin, and its maternal progenitor J. sabina var. sabina are genetically distinct with respect to cpDNA, SNP, and SilicoDArT loci. Mostly non-overlapping distributions of the sabina and balkanensis varieties are the result of their different climatic requirements. Context Juniperus sabina L. is present in the Eurasian mountains, but its range is severely fragmented. In Europe, two varieties of J. sabina occur: var. sabina and var. balkanensis, the latter being an allotetraploid hybrid between the diploid var. sabina and a tetraploid ancestor of Juniperus thurifera L. The distribution of the two varieties is mostly disjunct. Aims Assess the taxonomic affiliation and genetic differentiation of the populations of var. sabina and var. balkanensis in Europe and Asia using cpDNA, SilicoDArT, and SNP markers. Identify climatic niches of both juniper varieties in Europe. Methods Altogether, 21,134 SilicoDArT, 8,579 SNP, and four cpDNA loci were used. Seven climatic variables were compared in sites inhabited by var. balkanensis and the two parental species. Results The SilicoDArTs and SNPs revealed a pattern of population differentiation that was congruent with the cpDNA analysis. The hybrid var. balkanensis occupies habitats with higher temperatures and intermediate levels of precipitation compared to both parental taxa. Conclusion The low genetic variation and significant genetic differentiation among J. sabina populations likely result from the restriction of gene flow imposed by the mountain ranges. The balkanensis variety is able to cope with hot and dry climates probably thanks to the admixture of J. thurifera genes.