Genetic variation was examined for the endangered banded newts Triturus vittatus vittatus from a number of breeding sites in northern and central Israel near the southern limit of the species' distribution. Altitudes of the sites ranged from 15–740 m asl. Variations in nucleotide sequences of the mitochondrial cytochrome b gene (413-bp-long) and the control region (Dloop) (569-bp-long) were analyzed, of which the former varied at 13 nucleotide sites whereas the latter at 15 sites. Specimens from Berekhya pond, which is located at the lowest altitude, most distant from the other sites, and with the least annual precipitation, were found to be most divergent due to accumulation of anagenetic sequence changes. Sequence analysis showed that the newts from Israel are genetically collectively different from conspecific populations of Syria (Damascus) and Turkey (European part). DNA variation might have possibly been enhanced by differences in ecological conditions that are seemingly most prominent in habitats near the southern limit of the species' distribution.