1 Background: The Eneolithic (~ 5,500 yrBP) site of Verteba Cave in Western Ukraine contains 2 the largest collection of human skeletal remains associated with the archaeological 3 Cucuteni-Tripolye Culture. Their subsistence economy is based largely on agro-pastoralism and 4 had some of the largest and most dense settlement sites during the Middle Neolithic in all of 5 Europe. To help understand the evolutionary history of the Tripolye people, we performed 6 mtDNA analyses on ancient human remains excavated from several chambers within the cave.7Results: Burials at Verteba Cave are largely commingled and secondary in nature. A total of 68 8 individual bone specimens were analyzed. Most of these specimens were found in association 9 with well-defined Tripolye artifacts. We determined 28 mtDNA D-Loop (368 bp) sequences 10 and defined 8 sequence types, belonging to haplogroups H, HV, W, K, and T. These results do 11 not suggest continuity with local pre-Eneolithic peoples, but rather complete population 12 replacement. We constructed maximum parsimonious networks from the data and generated 13 population genetic statistics. Nucleotide diversity (π) is low among all sequence types and our 14 network analysis indicates highly similar mtDNA sequence types for samples in chamber G3.
15Using different sample sizes due to the uncertainly in number of individuals (11, 28, or 15), we 16 found Tajima's D statistic to vary. When all sequence types are included (11 or 28), we do not 17 find a trend for demographic expansion (negative but not significantly different from zero); 18 however, when only samples from Site 7 (peak occupation) are included, we find a significantly 19 negative value, indicative of demographic expansion.20 Conclusions: Our results suggest individuals buried at Verteba Cave had overall low mtDNA 21 diversity, most likely due to increased conflict among sedentary farmers and nomadic 22 pastoralists to the East and North. Early Farmers tend to show demographic expansion. We find 23 different signatures of demographic expansion for the Tripolye people that may be caused by 24 existing population structure or the spatiotemporal nature of ancient data. Regardless, peoples of 25 the Tripolye Culture are more closely related to early European farmers and lack genetic 26 continuity with Mesolithic hunter-gatherers or pre-Eneolithic groups in Ukraine.28 29 30 31 32 33 3 Background 1 Profound cultural transitions accompanied the Neolithic and Bronze Age in Europe. 2 These transitions were catalyzed initially through large-scale migrations of agriculturists from 3 the West Asia (modern day Anatolia) during the Neolithic, and later through the migrations of 4 steppe herders from the Pontic-Caspian and Volga regions of modern Ukraine during the 5 Bronze Age [1-4]. A continuing question of debate among researchers is how these migrations 6 affected the genetic composition of modern-day Europe [5, 6]. A key question in this debate is 7 whether these migrations that brought about cultural changes were the results of mo...