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Verteba Cave (VC) in western Ukraine dates to the Eneolithic period (c. 5500 YBP), and contains the largest collection yet found of human skeletal remains associated with the Cucuteni–Tripolye culture. The subsistence economy of this people was based on agropastoralism, and included some of the largest and densest Middle Neolithic settlement sites in all of Europe. To understand further the evolutionary history of the Tripolye people, we examined population genetics patterns in mitochondrial DNA from ancient human remains excavated from VC chambers. From five commingled and secondary burial sites within the cave, we obtained 368 bp mtDNA HVR 1 sequences from 22 individuals assignable to eight haplogroups: H (three haplotypes), HV (two haplotypes), W, K, and T. Overall nucleotide diversity is low (π = 0.00621). The two largest samples, from Chamber G3 and Site 7, were significantly differentiated with respect to haplotype composition: G3 (n = 8) is dominated by haplotype W (π = 0), whereas Site 7 (n = 15) is dominated by H haplotypes (π = 0.00439). Tajima’s D as an indication of population expansion was not significantly negative for the complete sample (D = –1.37) or for sites G3 (D = –0.973) and 7 (D = –1.35), which were analyzed separately. Individuals from the Tripolye culture buried at VC c. 5500 YBP had predominantly haplogroup H and related haplotypes. This contrasts with predominantly haplogroup U individuals in pre Eneolithic peoples from the same area, which suggests lack of genetic continuity in a site that has been dated to the Mesolithic. Peoples of the Tripolye culture are more closely related to other early European farmers than to Mesolithic hunter-gatherers and/or preEneolithic cultures.

Analysis of ancient human mitochondrial DNA from Verteba Cave, Ukraine: insights into the Late Neolithic-Chalcolithic Cucuteni–Tripolye culture