Multi-scale ancient DNA analyses confirm the western origin of Michelsberg farmers and document probable practices of human sacrifice

<div><p>In Europe, the Middle Neolithic is characterized by an important diversification of cultures. In northeastern France, the appearance of the Michelsberg culture has been correlated with major cultural changes and interpreted as the result of the settlement of new groups originating from the Paris Basin. This cultural transition has been accompanied by the expansion of particular funerary practices involving inhumations within circular pits and individuals in “non-conventional” positions (deposited in the pits without any particular treatment). If the status of such individuals has been highly debated, the sacrifice hypothesis has been retained for the site of Gougenheim (Alsace). At the regional level, the analysis of the Gougenheim mitochondrial gene pool (SNPs and HVR-I sequence analyses) permitted us to highlight a major genetic break associated with the emergence of the Michelsberg in the region. This genetic discontinuity appeared to be linked to new affinities with farmers from the Paris Basin, correlated to a noticeable hunter-gatherer legacy. All of the evidence gathered supports (i) the occidental origin of the Michelsberg groups and (ii) the potential implication of this migration in the progression of the hunter-gatherer legacy from the Paris Basin to Alsace / Western Germany at the beginning of the Late Neolithic. At the local level, we noted some differences in the maternal gene pool of individuals in "conventional" <i>vs</i>. "non-conventional" positions. The relative genetic isolation of these sub-groups nicely echoes both their social distinction and the hypothesis of sacrifices retained for the site. Our investigation demonstrates that a multi-scale aDNA study of ancient communities offers a unique opportunity to disentangle the complex relationships between cultural and biological evolution.</p></div

Localization of Gougenheim and of the sites anterior to 2,600 cal. BC. considered in the study.

<p>Central European groups (□); South European groups (○); French groups (Δ); MICH groups (◊). Hunter-gatherers (gray); Central European farmers (blue); South European farmers (red/orange); MNF farmers (pink); MICH farmers (green). Color gradient is given according to the chronology of each group. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179742#pone.0179742.s010" target="_blank">S6 Table</a> for details regarding the populations considered.</p

Principal component analysis (PCA) of the ancient mtDNA dataset.

<p>(a) PCA performed with haplogroup frequencies. (b) Circle of correlation. (c) Chronological overview of groups with the number of individuals between parentheses (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179742#pone.0179742.s010" target="_blank">S6 Table</a> for details). Colors are the same as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179742#pone.0179742.g002" target="_blank">Fig 2</a>. HGCN: Hunter-gatherers from Central Europe; HGW: Hunter-gatherers from West Europe; HGS: Hunter-gatherers from South Europe; STA: Starcevo-Körös; LBKT: Linearbandkeramik in Transdanubia; LBK: Linearbandkeramik; CRD: Cardial; EPI: EpiCardial Early Neolithic in Spain; RSG: Grossgartach/Planig-Friedberg/Rossen; MNF: Middle Neolithic in France; SCG: Schöningen; MICH: Michelsberg; BAC: Baalberge; LNS: Late Neolithic in Spain; SMZ: Salzmünde; BEC: Bernburg.</p

Spatial distribution of the mitochondrial (mtDNA) haplogroups within the Gougenheim site.

<p>(A) Inhumation repartition and mtDNA haplogroup distribution. Ellipses represent a circular pit; stars represent individuals in the "conventional position", whereas triangles represent individuals in the "non-conventional" position. (B) MtDNA haplogroup composition of "conventional" vs. "non-conventional" groups.</p

Pairwise <i>F</i>_ST distances.

<p><i>F</i>_ST measured between Gurgy and ancient European hunter-gatherers / farmer groups (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.s012" target="_blank">S4 Table</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.s018" target="_blank">S10 Table</a>). 'PRE_HG' for European hunter-gatherers anterior to 4,000 BC (N = 41), 'POST_HG' for European hunter-gatherers after 4,000 BC (N = 30), 'PRE_Central_F' for Central European Neolithic farmers anterior to 4,000 BC (N = 147), 'POST_Central_F' for Central European Neolithic farmers after 4,000 BC (N = 28), 'PRE_South_F' for Southern European Neolithic farmers anterior to 4,000 BC (N = 56), and 'POST_South_F' for Southern European Neolithic farmers after 4,000 BC (N = 49). Ancient groups anterior to 4,000 BC, which were sufficiently large for comparison at the population level are also shown (Derenburg, Halberstadt, Karsdorf, Salzmünde, from Germany [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref011" target="_blank">11</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref012" target="_blank">12</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref014" target="_blank">14</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref015" target="_blank">15</a>]; Los Cascajos from Spain [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref019" target="_blank">19</a>] and Les Treilles form southern France [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref054" target="_blank">54</a>]).</p

When the Waves of European Neolithization Met: First Paleogenetic Evidence from Early Farmers in the Southern Paris Basin

<div><p>An intense debate concerning the nature and mode of Neolithic transition in Europe has long received much attention. Recent publications of paleogenetic analyses focusing on ancient European farmers from Central Europe or the Iberian Peninsula have greatly contributed to this debate, providing arguments in favor of major migrations accompanying European Neolithization and highlighting noticeable genetic differentiation between farmers associated with two archaeologically defined migration routes: the Danube valley and the Mediterranean Sea. The aim of the present study was to fill a gap with the first paleogenetic data of Neolithic settlers from a region (France) where the two great currents came into both direct and indirect contact with each other. To this end, we analyzed the Gurgy 'Les Noisats' group, an Early/Middle Neolithic necropolis in the southern part of the Paris Basin. Interestingly, the archaeological record from this region highlighted a clear cultural influence from the Danubian cultural sphere but also notes exchanges with the Mediterranean cultural area. To unravel the processes implied in these cultural exchanges, we analyzed 102 individuals and obtained the largest Neolithic mitochondrial gene pool so far (39 HVS-I mitochondrial sequences and haplogroups for 55 individuals) from a single archaeological site from the Early/Middle Neolithic period. Pairwise <i>F</i>_ST values, haplogroup frequencies and shared informative haplotypes were calculated and compared with ancient and modern European and Near Eastern populations. These descriptive analyses provided patterns resulting from different evolutionary scenarios; however, the archaeological data available for the region suggest that the Gurgy group was formed through equivalent genetic contributions of farmer descendants from the Danubian and Mediterranean Neolithization waves. However, these results, that would constitute the most ancient genetic evidence of admixture between farmers from both Central and Mediterranean migration routes in the European Neolithization debate, are subject to confirmation through appropriate model-based approaches.</p></div

Location of the sites anterior to 4000 cal BC.

<p><b>PRE_HG</b> (purple): 1) La Braña, 2) La Pasiega, 3) La Chora, 4) Erralla, 5) Aizpea, 6) Oberkassel, 7) Reuland-Loschbour, 8) Bad Durrenberg, 9) Hohleinstein, 10) Hohler Fels, 11) Villabruna, 12) Paglicci, 13) Dolni Vestonice, 14) Spiginas, 15) Donkalnis, 16) Kretuonas, 17) Uznyi Oleni Ostrov, 18) Popovo, 19) Kostenki, 20) Chekalino, 21) Lebyazhinka. <b>PRE_South_F</b> (blue): 22) Los Cascajos, 23) Paternanbidea, 24) Chaves, 25) Can Sadurni, 26) Sant Pau del Camp, 27) Avellaner. <b>PRE_Central_F</b> (green): 28) Flomborn, 29) Wittmar, 30) Oberwiederstedt, 31) Unterwiederstedt, 32) Salzmünde, 33) Derenburg, 34) Halberstadt, 35) Naumburg, 36) Karsdorf, 37) Esperstedt, 38) Vedrovice, 39) Szarvas, 40) Ecsegfalva. References in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.s012" target="_blank">S4 Table</a>.</p

Principal Component Analysis (PCA) on the ancient mtDNA dataset.

<p>PCA performed with haplogroup frequencies and circle of correlation of the PCA. The ancient samples (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.s012" target="_blank">S4 Table</a>) included 'PRE_HG' for European Hunter Gatherers anterior to 4,000 BC (N = 41), 'PRE_Central_F' for Central European Neolithic farmers anterior to 4,000 BC (N = 147), and 'PRE_South_F' for Southern European Neolithic farmers anterior to 4,000 BC (N = 56). Some ancient groups anterior to 4,000 BC and sufficiently large enough for comparison at the population level are also shown (Derenburg, Halberstadt, Karsdorf, Salzmünde, from Germany [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref011" target="_blank">11</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref012" target="_blank">12</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref014" target="_blank">14</a>,<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref015" target="_blank">15</a>]; and Los Cascajos from Spain [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.ref019" target="_blank">19</a>]).</p

Multidimensional Scaling (MDS) on the ancient mtDNA dataset.

<p>MDS performed on the <i>F</i>_ST values between Gurgy and the ancient dataset (same as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0125521#pone.0125521.g001" target="_blank">Fig 1</a>).</p