Farming was first introduced to Europe in the mid-7th millennium BCE–associated with migrants from Anatolia who settled in the Southeast before spreading throughout Europe. To understand the dynamics of this process, we analyzed genome-wide ancient DNA data from 225 individuals who lived in southeastern Europe and surrounding regions between 12,000 and 500 BCE. We document a West-East cline of ancestry in indigenous hunter-gatherers and–in far-eastern Europe–early stages in the formation of Bronze Age Steppe ancestry. We show that the first farmers of northern and western Europe passed through southeastern Europe with limited hunter-gatherer admixture, but that some groups that remained mixed extensively, without the male-biased hunter-gatherer admixture that prevailed later in the North and West. Southeastern Europe continued to be a nexus between East and West, with intermittent genetic contact with the Steppe up to 2000 years before the migrations that replaced much of northern Europe’s population.

Farming was first introduced to southeastern Europe in the mid-7th millennium BCE – brought by migrants from Anatolia who settled in the region before spreading throughout Europe. To clarify the dynamics of the interaction between the first farmers and indigenous hunter-gatherers where they first met, we analyze genome-wide ancient DNA data from 223 individuals who lived in southeastern Europe and surrounding regions between 12,000 and 500 BCE. We document previously uncharacterized genetic structure, showing a West-East cline of ancestry in hunter-gatherers, and show that some Aegean farmers had ancestry from a different lineage than the northwestern Anatolian lineage that formed the overwhelming ancestry of other European farmers. We show that the first farmers of northern and western Europe passed through southeastern Europe with limited admixture with local hunter-gatherers, but that some groups mixed extensively, with relatively sex-balanced admixture compared to the male-biased hunter-gatherer admixture that prevailed later in the North and West. Southeastern Europe continued to be a nexus between East and West after farming arrived, with intermittent genetic contact from the Steppe up to 2,000 years before the migration that replaced much of northern Europe’s population.

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The cemetery is located in the south-west of Pottenbrunn, on plot "Steinfeld" (15°41´05"/48°13´55"). Discovered in 1930, it had already yielded objects dating to the early La Tène period. In 1981, road construction revealed further finds which initiated rescue excavations by the Bundesdenkmalamt (State Office for Protection of Historical Monuments) under the guidance of J.-W. Neugebauer (Ramsl 2002a(Ramsl , 13) in 1981(Ramsl and 1982. A total of 42 graves with 45 burials (single and double inhumations, and cremations) have been documented. Some burials were severely disturbed (by ancient activities such as grave robbing and/or contemporary construction work), and some were set within fenced enclosures ("Grabgärten"). Three (of 22) samples of charcoal and bone fragments taken by Peter Stadler (Department of Prehistory, Natural History Museum Vienna) in the course of the FWFproject "Absolute Chronology for Early Civilisations in Austria and Central Europe" returned AMS dates of 410-200 cal BCE (grave 520), 550-200 cal BCE (grave 565) and 380-350 cal BCE (grave 1005) (Ramsl 2002b, 359). The cremation burials were not included in the initial osteological analysis, but 31 inhumed individuals were studied (Gerold 2002). Petrous bones from three of these were successfully analyzed for aDNA. Sample I11699 (female) derived from an individual (inv. no. 26.238) aged c. 20 years in grave 89 which, despite disturbance in antiquity, was accompanied by fibulae and ceramic vessels. Sample I11701 (male) derived from an individual (inv. no. 26.249) aged c. 18 years in grave 570, which also included shears, fibulae, and ceramic vessels. Evidence for bone porosity in the mandible and maxilla suggest possible Vitamin C deficiency, while enamel hypoplasia points to malnutrition or illness during childhood. Sample I11708 (female) derived from an individual (inv.no. 26.250) aged c. 25-35 years in grave 574/2, who was richly adorned with fibulae, bronze, iron and silver-rings, an amber ring, a bracelet, a glass bead, and a worked bone artefact.