Long-distance kinship in megalithic Europe

Long-distance kinship in megalithic Europe | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Biological Sciences - Article Long-distance kinship in megalithic Europe Ben Krause-Kyora, Nicolas Antonio da Silva, Almut Nebel, Daniel Kolbe, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6464608/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Across Neolithic Europe, megalithic monuments – including stone circles and gallery graves – were constructed by early farming communities, representing a supra-regional cultural phenomenon1. The collective graves are often viewed as symbols of shared ancestry, social identity and regional connectivity. Palaeogenetic studies of megalithic burial sites have identified patrilineal kinship and locally restricted female mobility (≤ 8 km)2–5. However, until now, close genetic ties between geographically distant populations have not been described. Here we analyse genome-wide data from 203 individuals associated with five Wartberg and one Western Funnel Beaker communities. Our results reveal long-distance first- and second-degree kinship ties across these megalithic sites, involving both females and males. Remarkably, we identify close relatives — such as a father and son — who were buried more than 200 kilometres apart and belonged to distinct cultures. Additionally, we uncover extended genetic networks that link the six communities. Together, they form a single population with genetic boundaries to other megalithic societies. Our findings suggest that mobility and mating networks spanned hundreds of kilometres, fostering strong intra-group cohesion while maintaining limited external contacts. This indicates that the supra-regional megalithic phenomenon may not reflect deep social integration, but rather a shared cultural expression without strong underlying biological connections. Biological sciences/Genetics/Genomics/Genome evolution Biological sciences/Evolution/Archaeology Biological sciences/Evolution/Anthropology/Biological anthropology Biological sciences/Genetics/Population genetics Figures Figure 1 Figure 2 Figure 3 Figure 4 Main From the Iberian Peninsula to Scandinavia, megalithic monuments formed a large-scale Neolithic phenomenon (ca. 4500-2800 BCE). Various agricultural groups constructed dolmens, passage graves, stone circles and avenues, among other structures 6,7 . While these monuments express distinct local and regional identities, they also signify broader supra-regional networks that connected communities across vast distances 1 . One of these farming communities in central Europe is represented by the Late Neolithic Wartberg culture (WBC; 3500-2800 BCE), spreading across the western low mountain range of present-day Germany (Figure 1) 8–10 . WBC is renowned for its subterranean gallery tombs containing up to 350 people and the oldest wall engravings of wagons in Europe 10 . WBC individuals are characterized by a high western hunter-gatherer (WHG) ancestry. A plausible scenario is that the WBC population formed through the admixture of Middle Neolithic Michelsberg farmers and local WHG groups 11,12 . Another contemporaneous megalithic society near the WBC is the Western Funnel Beaker Group (Trichterbecherkultur-West, TRB-West; Figure 1). Archaeological evidence suggests that the TRB-West population also originated from Middle Neolithic farmers, most likely with influences from the Michelsberg context 13 . However, to date, no genomic data of TRB-West individuals has been analysed, leaving questions about their genetic affinities and possible kinship relations with the WBC and other megalithic societies unanswered. Previous research on megalithic communities from Sweden, Ireland, England and France have reconstructed family trees and kinship networks, revealing a pattern of predominantly patrilineal descent 2–5 and female mobility within a radius of up to 8 km 2 . Population genetic structure We analysed genomes of 203 individuals, including 129 newly generated genomes, from five WBC (Altendorf, Niedertiefenbach, Rimbeck, Warburg, Züschen) and one TRB-West site (Sorsum) (Figure 1, Table 1, Supplementary Table 1; Supplementary Note 1). The samples represent a large proportion of all WBC-associated human remains uncovered so far 9 . Interestingly, principal component analysis reveals that the individuals from Sorsum cluster with the five WBC sites, rather than with those from the TRB-North or -East subgroups (Figure 2). In addition, the Sorsum individuals exhibit a high level of WHG ancestry (25-48%; mean: 33%) that falls within the range observed for WBC (22-56%; mean: 35%). They also show a mode and date of admixture (between Middle Neolithic farmers and WHG) that are similar to those of WBC populations (Supplementary Note 2; Supplementary Figures 1-10; Supplementary Table 2). Close kinship relationships across sites Given the close genetic affinities between Sorsum and the WBC populations, we investigated kinship relations within and across sites using a conservative approach that integrates results from three independent methods (READv2, KIN, and NGSRelate2) 14–16 . Surprisingly, we identified six cross-site relationships, five of which involve individuals from Sorsum, including a first-degree kinship between a father (NT30, Niedertiefenbach) and a subadult son (361.0201/b, Sorsum) (Figure 3; Supplementary Table 3). Sorsum is connected to both Niedertiefenbach and Warburg via two second-degree relationships each. The other cross-site kinship pair (second-degree) links Niedertiefenbach with Warburg. Sorsum is highly connected and lies in an area with contact between WBC and TRB-West (Figure 1). It is located approximately 225 km from Niedertiefenbach, making it the furthest distance between any of the six sites. It is also the longest distance separating first- and second-degree relatives reported for the Neolithic to date. Noteworthy, Sorsum and Niedertiefenbach predominantly consist of males carrying the I2c Y chromosome lineage, which also includes the father-son pair mentioned above (Figure 3; Supplementary Table 1). Out of the eleven individuals connected by the six close relationships, five are females, suggesting that both males and females moved across sites. Sorsum and WBC as one population Genotype imputation was applied for individuals with sufficient genomic coverage (n=682, including published contemporaneous samples) to estimate identity-by-descent (IBD) segments using ancIBD 17 (Supplementary Table 4; Supplementary Figure 11). We observed no strong IBD sharing (sum of IBD > 20cM) between Sorsum (TRB-West) and other TRB (North/East) populations (Supplementary Table 4). However, Sorsum and the five WBC sites share more IBD segments amongst each other than with any other Late Neolithic European group including megalithic communities (Figure 4; Supplementary Figure 12). In summary, the Sorsum and WBC communities are part of a single population, as demonstrated by their close genetic affinities in multiple analyses (Supplementary Note 2; Supplementary Figures 1-10) and direct kinship connections (Figure 3-4). Our results indicate that individuals from the WBC sites and Sorsum predominantly chose partners from within this single population rather than integrating people from the outside, thus forming a cohesive reproductive unit and maintaining strict genetic boundaries, likely through social and/or cultural norms. One possible explanation for their close connection is that Sorsum was the northern most location of a WBC site, adopting some TRB material culture while staying in contact with the WBC populations. Alternatively, TRB-West and WBC may have had a common origin, with Sorsum preserving genetic continuity and exchange with WBC over time, despite cultural differences. No close ties between WBC/Sorsum and other megalithic groups WBC/Sorsum exhibit their closest external relationship (60-75 cM, likely above fourth degree) with samples from the Blätterhöhle cave in northwestern Germany, who in turn are connected to outliers (characterized by high WHG ancestry) from Mont Aimé in the Paris Basin (Figures 2,4; Supplementary Table 4). The large number of IBD segments and Y chromosome haplogroups shared between the individuals implies that their high WHG ancestry may be due to a common genetic origin (i.e., they had the same WHG ancestors). Similarly, when considering additional published contemporaneous megalithic groups, we detected two other large clusters of individuals sharing IBD segments (Supplementary Figure 12). These two clusters are in England/Scotland and Sweden, respectively, and do not show connections to each other or to our Sorsum/WBC network. These findings indicate that while megalithic societies may have shared architectural styles and certain cultural practices, genetic exchange through direct kinship did not extend across vast distances or across the English Channel and the Baltic Sea. Instead, kinship networks appear to have been maintained within more localized or regionally structured populations. Intra-site kinship relationships Within the six communities analysed here, we identified 123 closely related pairs, comprising 44 first-degree and 79 second-degree relationships (Figure 3; Supplementary Table 3). In addition, we detected 39 third-degree related pairs (Supplementary Figure 13; Supplementary Table 3). About half (54%) of the 203 individuals have a close relative (first- or second-degree). This proportion is comparatively low when considering other megalithic populations 2,3,5 . In total, we identified 19 clusters of close relatives. Most clusters are relatively small, ranging from isolated pairs to groups of up to six individuals. Larger family networks are seen in Niedertiefenbach (12 individuals), Sorsum (two clusters with 11 and 20 individuals) and Warburg (13 individuals). In the Warburg necropolis, the family ties span two of the three burial complexes (Warburg I and Warburg III) 18 —indicating that they were used contemporaneously. The presence of single pairs or small clusters of individuals who are not closely related to the four big families could indicate that burial in the chambers was not restricted to them. Instead, WBC and TRB-West may have emphasized broader community membership, social affiliation or other forms of non-kin inclusion in their funerary practices. Patrilineal inheritance without constraints on female mobility To investigate sex-specific mobility patterns within WBC and Sorsum, we examined pairwise mismatches between same-sex pairs (male-male and female-female). Our results reveal no significant differences in relatedness between male and female pairs (Supplementary Figure 14), suggesting a relatively egalitarian mobility for the two sexes. This pattern contrasts with studies on other megalithic groups, such as those from the TRB-North or the British and Irish Neolithic, which show evidence for patrilocality with exogamous marriage practices 2,3 . Interestingly, despite this lack of strict patrilocality in WBC and Sorsum, we observed dominant paternal lineages within each community. The Y chromosome haplogroup I2c is prevalent in Niedertiefenbach, Altendorf and Sorsum, whereas I2a is most common in Warburg (Figures 2-3; Supplementary Table 1). This pattern suggests that the men involved in founding the different communities had distinct I lineages, which remained predominant over time. However, rather than suggesting dynastic rule or rigid male inheritance structures, it may reflect communal burial traditions where individuals were interred based on shared ancestry or social affiliation and not hierarchical status. Notably, females are underrepresented in all WBC and Sorsum sites, comprising only about 40% of all individuals (Supplementary Figure 15). Females might have been buried in different locations due to social status or ritual customs. A similar underrepresentation of females has also been observed in other megalithic grave communities with strong patrilinear family lineages 2–4 . Conclusion Our findings reveal a complex interaction pattern in central Europe during the Late Neolithic. The presence of first- and second-degree relationships spanning several sites, especially the father-son pair bridging Niedertiefenbach (WBC) with Sorsum (TRB-West), indicates that there was substantial contact between megalithic communities that were not only far apart (over 200 km), but also had cultural differences in ceramic styles. Their close relation is further substantiated by the archaeological record which shows similarities in burial construction (rock-cut chamber of Sorsum and WBC gallery graves) 19 and artefacts (hunting equipment—such as transverse and triangular arrowheads and bone points) 20–22 . Collectively our evidence points towards a much higher mobility than previously thought 2,23 . Despite this high mobility, WBC and Sorsum lacked close kinship ties with other megalithic populations, revealing restricted mating practices. These observations add an important layer to our understanding of megalithic societies. Although the use of large blocks and collective burial practices is a common feature across regions with megalithic structures, our findings also reveal notable differences, as reflected in the varying spatial ranges of kinship ties among the buried individuals. These differences imply that the same social sphere may have held distinct meanings for the various groups. In conclusion, our results point not only to a spatial dimension of kinship relations in the Neolithic but also to the complex nature of the megalithic phenomenon. Methods Inclusion & ethics statement We followed the guidelines for ethical ancient DNA (aDNA) research on human remains 24 . Sampling, DNA extraction, library preparation and whole genome sequencing The sampling and DNA extraction procedures were carried out in a specialized ancient DNA facility at Kiel University, as outlined previously 25 . We collected a total of 479 commingled skeletal elements from four Late Neolithic collective burial sites located in present-day Germany: Altendorf, Sorsum, Warburg and Züschen. Some individuals are represented by more than one skeletal element. The DNA was extracted from bone fragments (specific bone source for each sample that entered our analysis is detailed in Supplementary Table 1) and converted into partial uracil-DNA glycosylase (UDG) double-stranded libraries 26 . The libraries were submitted for whole genome shotgun sequencing on the Illumina HiSeq 6000 (2x100) platform at the Institute of Clinical Molecular Biology in Kiel. Raw read processing and ancient DNA authentication Adapter sequences were trimmed and paired-end reads were merged using ClipAndMerge version 1.7.7 27 . The resulting reads were then aligned to the human reference genome (build hg19) using BWA version 0.7.15 28 , with relaxed mapping stringency (flag -n 0.01) to account for the expected mismatches in aDNA. If published and newly generated datasets were available for a given individual, they were merged into a single BAM file with Samtools v1.12 29 . Duplicate reads were removed using DeDup version 0.12.1 27 . All samples were evaluated for deamination with DamageProfiler version 1.1 30 . Additionally, contamination was estimated based on mitochondrial DNA (mtDNA) using Schmutzi version 1.5.5.5 31 and the X chromosome in male samples using ANGSD version 0.935 32 . Generation of pseudo-haploid genotype datasets The two terminal positions of reads in the BAM files were trimmed using bamUtil version 1.0.15 33 to decrease the impact of aDNA deamination prior to genotyping. Next, we used SequenceTools (https://github.com/stschiff/sequenceTools) version 1.2.2 to create pseudo-haploid genotypes at positions from the 1240K and Human Origins (HO) panels. Samples with fewer than 20,000 SNPs covered in the 1240K panel were removed from population genetic analyses. The filtered genotype dataset comprising 129 samples was integrated with the Allen Ancient DNA Resource (AADR) reference panels (version 62.0 of the 1240K and HO panels) – a comprehensive collection of published aDNA data 34 . In addition, we incorporated previously published data from Altendorf (n = 15) and Warburg (n = 17), along with samples from Niedertiefenbach (n = 40) and Rimbeck (n = 2) 11,12 . This brought the total number of individuals associated with the WBC and Sorsum communities analysed in this study to 203 (Table 1). Inference of genetic sex and uniparental haplogroups The genetic sex of the samples was determined using two methods that assess the ratio of sequences aligning to sex chromosomes relative to autosomes 35,36 . The mtDNA haplogroup of each sample was inferred with HaploGrep2 version 2.4.0 37 . For male individuals, Y chromosome haplogroups were determined using yHaplo v1.1.2 38 , based on either the full set of ISOGG SNPs or a filtered set restricted to transversions only. The haplogroup assignment with the highest resolution was retained for each individual. For the genetic sex determination and uniparental haplogroup inference, a mapping and base quality threshold of 30 was applied. Ancestry inference: PCA, f-statistics and admixture modelling A principal component analysis (PCA) was performed on the merged HO dataset with smartpca version 16000 39 using the option “lsqproject: YES”. The ancient samples were projected onto the genetic variation of modern West Eurasian populations from the HO panel. We computed f4 statistics with qpDstat version 970 40 with the option “f4mode: YES”. In the f4 analyses (results described inSupplementary Note 2), the 1240K dataset was used, and the Mbuti was included as the outgroup. We conducted a hypothesis-free ancestry deconvolution analysis using ADMIXTURE version 1.3.0 41 in unsupervised mode, exploring 3 to 12 components with 200 bootstraps (Supplementary Note 2). The analysis was performed on a pruned HO dataset using PLINK version 1.90b6.21 with the command “--indep-pairwise 200 25 0.4” 42 . Hypothesis-driven admixture modelling with two sources was performed with qpAdm version 1201 40 using the option “allsnps: NO” (Supplementary Note 2). The set of sources and outgroups used for qpAdm modelling is available in the Supplementary Table 2header. Additionally, we used DATES version 753 43 to model the most recent time of admixture, considering a single pulse that led to the formation of Sorsum and the WBC communities (Supplementary Note 2). The following parameters were used in DATES: 'binsize: 0.001’, 'maxdis: 1.0’, 'seed: 77’, 'jackknife: YES’, 'qbin: 10’, 'runfit: YES’, 'afffit: YES’, 'lovalfit: 0.45’, 'minparentcount: 1’. Assessing sex-biased admixture We assessed the possibility of sex-biased admixture by estimating the ancestry proportions on autosomes and the X chromosome (results in Supplementary Note 2). To this end, we relied on the 1240K SNP panel, which we initially pruned using PLINK version 1.90b6.21 (command “--indep-pairwise 200 25 0.4”) 42 . After linkage disequilibrium pruning, 569,582 autosomal and X chromosomal 6,737 SNPs remained that were subsequently used for supervised ADMIXTURE modelling (version 1.3.0) 41 . Only samples with sufficient coverage for at least 1000 X chromosomal SNPs were included in the analysis. Due to the small number of variants available on the X chromosome, we resampled the autosomal SNPs 1000 times and matched the number of X chromosomal SNPs. For each individual, we calculated the ratio of WHG ancestry on the X chromosomes to the WHG ancestry on autosomes (R X/A ) and performed a log 2 transformation of this statistic to better visualize deviations from the null hypothesis (i.e., no imbalance between the ancestral proportions of autosomes and X chromosomes). A Wilcoxon signed-rank test was performed to determine whether the differences in estimated WHG ancestry between autosomes and X chromosomes were statistically significant 44 . Kinship analysis for Sorsum and the WBC communities To assess kinship among individuals from Sorsum and the WBC sites, we used the following three programs: READv2, KIN, and NGSRelate2 14–16 . As we observed a difference in the baseline mismatch rate between pairs from Sorsum (0.274) and those from WBC populations (0.252) (Supplementary Figure 16), which may lead to under- or overestimation of relatedness, we conducted separate analyses to ensure accurate kinship assessment. First, we only considered Sorsum samples to identify kinship within this population. Second, we only analysed WBC samples to assess kinship exclusively within this group. Finally, we combined samples from both Sorsum and WBC sites to identify kinship between the two populations. The analyses were conducted using SNPs from the 1240K panel, applying the following filters: kinship estimates were disregarded for a pair if fewer than 2,000 shared SNPs were available (for READv2 and NGSRelate2) or if the log-likelihood ratio was below 1 (for KIN). All three methods yielded highly consistent results, attesting to the reliability of the estimates (Supplementary Figure 17). To conservatively construct the close kinship network, we used a majority-rule consensus approach, where kinship assignments were determined based on agreement from at least two of the three methods. For example, if one program classified a pair as first-degree relatives while the other two assigned them as second-degree, we considered them second-degree kin. Pairs for which all three methods produced conflicting results were excluded from the kinship network. To ensure robustness, we primarily focused on first- and second-degree relationships as kinship estimates beyond this range carry greater uncertainty. For completeness, third-degree kinship estimates were also included in the Supplementary Information (Supplementary Figure 13; Supplementary Table 3). Identifying IBD segments We employed ancIBD version 0.3a1 17 to identify identity-by-descent (IBD) segments. These segments were primarily utilized to explore more distant relationships between individuals from Sorsum, WBC and other contemporaneous groups in the region (4000 – 2000 BCE; Supplementary Table 4). Additionally, the ancIBD results were also used to validate the kinship established through the majority-rule consensus method outlined above, whenever possible. For this analysis, we first imputed data from high-coverage ancient genomes (mean depth ≥ 0.1X) using GLIMPSE version 2.0.0 45 . Imputation was done with the 1000 Genomes Project reference panel. ancIBD was run with the default parameters. Since most biological relatives up to the sixth degree share at least two long IBD segments 46 , we included only pairs with three or more IBD segments larger than 12 cM in the IBD networks. Pedsim version 1.4 46 was used to simulate IBD segments across several kinship scenarios for a comparison against the IBD estimates observed in the ancient populations. Estimating ROH and N e We analysed runs of homozygosity (ROH) and effective population size (Ne) using HapROH version 0.3a4 47 on high-coverage samples (≥ 400,000 SNPs from the 1240K panel covered) from Sorsum, WBC and other published groups between 4000 and 2000 BCE (results described in Supplementary Note 2). The program was executed with default parameters. Declarations Data availability All the sequenced data generated in this study is deposited to the European Nucleotide Archive (accession: PRJEB88326). Acknowledgements This study was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) through the CRC 1266 – project number 290391021 and the Clusters of Excellence ROOTS (EXC 2150 ID 390870439). Author contributions B.K.-K. and J.Mü. designed the research. F.K., I.G., K.S. and M.R. provided the archaeological materials and context information. Laboratory work was supervised by B.K.-K. N.A.d.S. primarily performed the analyses, with assistance from D.K. and D.A.M. Ch.R., J.Me. and J.Mü. contributed to embedding the findings into the broader chronological and archaeological framework. K.F. and Ch.M. curated the samples. N.A.d.S., A.N., J.Mü. and B.K.-K. interpreted the results and wrote the manuscript, with all the authors reviewing. A.N. and B.K.-K. secured funding and supervised the project. Competing interests The authors declare no competing interests Materials & Correspondence Correspondence to Ben Krause-Kyora ( [email protected] ) References Schulz Paulsson, B. Time and Stone : The Emergence and Development of Megaliths and Megalithic Societies in Europe . (Archaeopress Publishing, Oxford, 2017). Seersholm, F. V. et al. Repeated plague infections across six generations of Neolithic Farmers. Nature 632 , 114–121 (2024). Fowler, C. et al. A high-resolution picture of kinship practices in an Early Neolithic tomb. Nature 601 , 584–587 (2022). Cassidy, L. M. et al. 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Parental relatedness through time revealed by runs of homozygosity in ancient DNA. Nat. Commun. 12 , 5425 (2021). Rinne, C., Drummer, C. & Hamann, C. Collective and individual burial practices. Changing patterns at the beginning of the third millennium BC: The megalithic grave of Altendorf. J. Neolit. Archaeol. 75–88 (2019) doi:10.12766/jna.2019.4. Tables Table 1. Overview of archaeological sites analysed in this study. N: number of individuals with whole genome sequencing data. Archaeological site Dating (BCE) N (previously published) N (newly generated) N total Altendorf 3250 – 3100 48 15 13 28 Niedertiefenbach 3350 – 3250 40 - 40 Rimbeck 3350 – 3075 * 2 - 2 Sorsum 3350 – 3100 - 56 56 Warburg 3350 – 2950 17 54 71 Züschen 3500 – 3150 - 6 6 Total 74 129 203 * Data provided as personal communication from C. Rinne. Additional Declarations There is NO Competing Interest. Supplementary Files Supplementarytables.xlsx Supplementary Table 1-4 WartbergSI.docx Supplementary Information Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6464608","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":452279355,"identity":"2287e6b9-e239-4d9c-bf9f-ed3865b1adb1","order_by":0,"name":"Ben Krause-Kyora","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0001-9435-2872","institution":"Kiel University","correspondingAuthor":true,"prefix":"","firstName":"Ben","middleName":"","lastName":"Krause-Kyora","suffix":""},{"id":452279356,"identity":"63fba978-1c7c-47fb-b883-4a277d7e3ffe","order_by":1,"name":"Nicolas Antonio da Silva","email":"","orcid":"https://orcid.org/0000-0002-1537-3286","institution":"Kiel University","correspondingAuthor":false,"prefix":"","firstName":"Nicolas","middleName":"Antonio da","lastName":"Silva","suffix":""},{"id":452279357,"identity":"0948b3a3-7226-4003-a847-bfd362d737dd","order_by":2,"name":"Almut Nebel","email":"","orcid":"","institution":"Christian-Albrechts-University","correspondingAuthor":false,"prefix":"","firstName":"Almut","middleName":"","lastName":"Nebel","suffix":""},{"id":452279358,"identity":"5333ba64-2d69-406e-86f7-b5ff8bb1646f","order_by":3,"name":"Daniel Kolbe","email":"","orcid":"","institution":"Kiel University","correspondingAuthor":false,"prefix":"","firstName":"Daniel","middleName":"","lastName":"Kolbe","suffix":""},{"id":452279359,"identity":"1ab6550e-a794-4496-900e-f16a4553a472","order_by":4,"name":"Daniel Myburgh","email":"","orcid":"https://orcid.org/0009-0006-2380-7490","institution":"Kiel University","correspondingAuthor":false,"prefix":"","firstName":"Daniel","middleName":"","lastName":"Myburgh","suffix":""},{"id":452279360,"identity":"86798078-3c87-40ed-876a-6f0ba594016b","order_by":5,"name":"Florian Klimscha","email":"","orcid":"","institution":"Georg-August-Universität zu Göttingen \u0026 Nds. Landesmuseum Hannover","correspondingAuthor":false,"prefix":"","firstName":"Florian","middleName":"","lastName":"Klimscha","suffix":""},{"id":452279361,"identity":"1d7f2aac-f512-47cd-9114-34d505267057","order_by":6,"name":"Irina Görner","email":"","orcid":"","institution":"Hessen Kassel Heritage","correspondingAuthor":false,"prefix":"","firstName":"Irina","middleName":"","lastName":"Görner","suffix":""},{"id":452279362,"identity":"51b7e67b-6fc5-4572-ba40-7fa96433f0f2","order_by":7,"name":"Katharina Fuchs","email":"","orcid":"https://orcid.org/0000-0003-0570-9337","institution":"Kiel University","correspondingAuthor":false,"prefix":"","firstName":"Katharina","middleName":"","lastName":"Fuchs","suffix":""},{"id":452279363,"identity":"666a029b-3b4c-45d2-bf50-349dd4ea6744","order_by":8,"name":"Christian Meyer","email":"","orcid":"","institution":"Kiel University","correspondingAuthor":false,"prefix":"","firstName":"Christian","middleName":"","lastName":"Meyer","suffix":""},{"id":452279364,"identity":"efcf30ba-9db8-4fce-9016-3a4fe052a7e1","order_by":9,"name":"Kerstin Schierhold","email":"","orcid":"","institution":"LWL-Archäologie für Westfalen","correspondingAuthor":false,"prefix":"","firstName":"Kerstin","middleName":"","lastName":"Schierhold","suffix":""},{"id":452279365,"identity":"5c859497-82f4-4739-9f2c-e1bba67cd9b8","order_by":10,"name":"Michael Rind","email":"","orcid":"","institution":"LWL-Archäologie für Westfalen","correspondingAuthor":false,"prefix":"","firstName":"Michael","middleName":"","lastName":"Rind","suffix":""},{"id":452279366,"identity":"2805ce73-eaf8-443a-b2ff-4344bbdef30c","order_by":11,"name":"John Meadows","email":"","orcid":"https://orcid.org/0000-0002-4346-5591","institution":"Centre for Baltic and Scandinavian Archaeology","correspondingAuthor":false,"prefix":"","firstName":"John","middleName":"","lastName":"Meadows","suffix":""},{"id":452279367,"identity":"01cc2b89-e2a4-4fa6-b4a3-ce7edaf3f729","order_by":12,"name":"Christoph Rinne","email":"","orcid":"https://orcid.org/0000-0002-9829-6182","institution":"Kiel University, Institute of Pre- and Protohistoric Archaeology","correspondingAuthor":false,"prefix":"","firstName":"Christoph","middleName":"","lastName":"Rinne","suffix":""},{"id":452279368,"identity":"cced7e2b-8d05-49a3-a20b-8d7fb9d5965d","order_by":13,"name":"Johannes Müller","email":"","orcid":"https://orcid.org/0000-0002-3538-197X","institution":"Institute of Pre- and Protohistoric Archaeology, Kiel University","correspondingAuthor":false,"prefix":"","firstName":"Johannes","middleName":"","lastName":"Müller","suffix":""}],"badges":[],"createdAt":"2025-04-16 14:40:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6464608/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6464608/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":82129036,"identity":"44ff4a3b-0fbf-4ebd-8c99-2bfb3c07df45","added_by":"auto","created_at":"2025-05-07 05:00:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":91850,"visible":true,"origin":"","legend":"\u003cp\u003eMap of north-western Europe showing the locations of the six archaeological sites analysed in this study. The broader distribution areas of WBC and TRB-West to which these sites belong are shown. Other relevant archaeological groups are highlighted. Additionally, the locations of characteristic megalithic structures are marked. The sites of Blätterhöhle and Mont Aimé are also indicated.\u003c/p\u003e","description":"","filename":"Fig1.MainMap.png","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/4fe6de8713ded4c2bebdf97c.png"},{"id":82128587,"identity":"b51a63fa-6bbd-4a09-ae9d-50744f74e7a8","added_by":"auto","created_at":"2025-05-07 04:52:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":220225,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e,\u003cstrong\u003e \u003c/strong\u003ePrincipal component analysis (PCA) of ancient individuals projected onto the genetic variation of present-day West Eurasian populations (light grey). \u003cstrong\u003eb\u003c/strong\u003e, Zoomed-in view of the space occupied by individuals from WBC and TRB communities. Samples from Blätterhöhle and Mont Aimé are highlighted.\u003c/p\u003e","description":"","filename":"Fig2.PCAv2.png","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/16e58646540ef850df376e45.png"},{"id":82128588,"identity":"d7f79301-1c54-4d31-b1de-918df6221597","added_by":"auto","created_at":"2025-05-07 04:52:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":61502,"visible":true,"origin":"","legend":"\u003cp\u003eKinship network of individuals identified as first- and second-degree relatives. Each node represents an individual, and the edge width denotes the degree of the genetic relationship. Clusters are positioned approximately according to the geographic locations of their respective sites, illustrating the spatial distribution of close kinship ties across the region. Male individuals are depicted with coloured borders indicating their Y chromosome haplogroups. Single letters mark the precise location of the sites.\u003c/p\u003e","description":"","filename":"Fig3.Kinshipmap.png","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/808be35d4c587ed9cf0949a1.png"},{"id":82128585,"identity":"395f0f65-d81e-421c-bee8-216e2395e6d4","added_by":"auto","created_at":"2025-05-07 04:52:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":102413,"visible":true,"origin":"","legend":"\u003cp\u003eIdentity-by-descent (IBD) network showing genetic connections between individuals analysed in this study and previously published samples. Each node represents an individual, with the edge width indicating the maximum shared IBD segment length (in centimorgans, cM). Nodes are positioned roughly according to the geographic locations of their respective sites, illustrating the spatial distribution of genetic ties. Single letters mark the precise location of the sites. Individuals from Blätterhöhle and Mont Aimé who are directly or indirectly connected to members of the six burial communities are shown. We display only those pairs related up to approximately the sixth degree (i.e., those sharing at least three IBD segments \u0026gt; 12 cM). Male individuals are outlined with coloured borders indicating their Y chromosome haplogroups.\u003c/p\u003e","description":"","filename":"Fig4.IBDmap.png","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/0d828bd1af24acb52c713a6b.png"},{"id":84160354,"identity":"468959ef-612b-4708-9ec9-369f9ef7553f","added_by":"auto","created_at":"2025-06-08 13:49:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1379728,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/5ac68f07-1602-4d9e-9690-40d43f2c28ad.pdf"},{"id":82128590,"identity":"5de47456-22bc-42ff-bded-5beceddf6118","added_by":"auto","created_at":"2025-05-07 04:52:39","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1299678,"visible":true,"origin":"","legend":"Supplementary Table 1-4","description":"","filename":"Supplementarytables.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/3925875ee0dc98d2be4d088f.xlsx"},{"id":82128589,"identity":"c4e80815-1fb3-439c-9e89-4cb14a5530a5","added_by":"auto","created_at":"2025-05-07 04:52:39","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":1295313,"visible":true,"origin":"","legend":"Supplementary Information","description":"","filename":"WartbergSI.docx","url":"https://assets-eu.researchsquare.com/files/rs-6464608/v1/f5437ff474046ecd39a81365.docx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Long-distance kinship in megalithic Europe","fulltext":[{"header":"Main","content":"\u003cp\u003eFrom the Iberian Peninsula to Scandinavia, megalithic monuments formed a large-scale Neolithic phenomenon (ca. 4500-2800 BCE). Various agricultural groups constructed dolmens, passage graves, stone circles and avenues, among other structures\u003csup\u003e6,7\u003c/sup\u003e. While these monuments express distinct local and regional identities, they also signify broader supra-regional networks that connected communities across vast distances\u003csup\u003e1\u003c/sup\u003e. One of these farming communities in central Europe is represented by the Late Neolithic Wartberg culture (WBC; 3500-2800 BCE), spreading across the western low mountain range of present-day Germany (Figure 1)\u003csup\u003e8–10\u003c/sup\u003e. WBC is renowned for its subterranean gallery tombs containing up to 350 people and the oldest wall engravings of wagons in Europe\u003csup\u003e10\u003c/sup\u003e. WBC individuals are characterized by a high western hunter-gatherer (WHG) ancestry. A plausible scenario is that the WBC population formed through the admixture of Middle Neolithic Michelsberg farmers and local WHG groups\u003csup\u003e11,12\u003c/sup\u003e. Another contemporaneous megalithic society near the WBC is the Western Funnel Beaker Group (Trichterbecherkultur-West, TRB-West; Figure 1). Archaeological evidence suggests that the TRB-West population also originated from Middle Neolithic farmers, most likely with influences from the Michelsberg context\u003csup\u003e13\u003c/sup\u003e. However, to date, no genomic data of TRB-West individuals has been analysed, leaving questions about their genetic affinities and possible kinship relations with the WBC and other megalithic societies unanswered. Previous research on megalithic communities from Sweden, Ireland, England and France have reconstructed family trees and kinship networks, revealing a pattern of predominantly patrilineal descent\u003csup\u003e2–5\u003c/sup\u003e and female mobility within a radius of up to 8 km\u003csup\u003e2\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePopulation genetic structure\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe analysed genomes of 203 individuals, including 129 newly generated genomes, from five WBC (Altendorf, Niedertiefenbach, Rimbeck, Warburg, Züschen) and one TRB-West site (Sorsum) (Figure 1, Table 1, Supplementary Table 1; Supplementary Note 1). The samples represent a large proportion of all WBC-associated human remains uncovered so far\u003csup\u003e9\u003c/sup\u003e. Interestingly, principal component analysis reveals that the individuals from Sorsum cluster with the five WBC sites, rather than with those from the TRB-North or -East subgroups (Figure 2). In addition, the Sorsum individuals exhibit a high level of WHG ancestry (25-48%; mean: 33%) that falls within the range observed for WBC (22-56%; mean: 35%). They also show a mode and date of admixture (between Middle Neolithic farmers and WHG) that are similar to those of WBC populations (Supplementary Note 2; Supplementary Figures 1-10; Supplementary Table 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClose kinship relationships across sites\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGiven the close genetic affinities between Sorsum and the WBC populations, we investigated kinship relations within and across sites using a conservative approach that integrates results from three independent methods (READv2, KIN, and NGSRelate2)\u003csup\u003e14–16\u003c/sup\u003e. Surprisingly, we identified six cross-site relationships, five of which involve individuals from Sorsum, including a first-degree kinship between a father (NT30, Niedertiefenbach) and a subadult son (361.0201/b, Sorsum) (Figure 3; Supplementary Table 3). Sorsum is connected to both Niedertiefenbach and Warburg via two second-degree relationships each. The other cross-site kinship pair (second-degree) links Niedertiefenbach with Warburg. Sorsum is highly connected and lies in an area with contact between WBC and TRB-West (Figure 1). It is located approximately 225 km from Niedertiefenbach, making it the furthest distance between any of the six sites. It is also the longest distance separating first- and second-degree relatives reported for the Neolithic to date. Noteworthy, Sorsum and Niedertiefenbach predominantly consist of males carrying the I2c Y chromosome lineage, which also includes the father-son pair mentioned above (Figure 3; Supplementary Table 1). Out of the eleven individuals connected by the six close relationships, five are females, suggesting that both males and females moved across sites.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSorsum and WBC as one population\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGenotype imputation was applied for individuals with sufficient genomic coverage (n=682, including published contemporaneous samples) to estimate identity-by-descent (IBD) segments using ancIBD\u003csup\u003e17\u003c/sup\u003e (Supplementary Table 4; Supplementary Figure 11). We observed no strong IBD sharing (sum of IBD \u0026gt; 20cM) between Sorsum (TRB-West) and other TRB (North/East) populations (Supplementary Table 4). However, Sorsum and the five WBC sites share more IBD segments amongst each other than with any other Late Neolithic European group including megalithic communities (Figure 4; Supplementary Figure 12).\u003c/p\u003e\n\u003cp\u003eIn summary, the Sorsum and WBC communities are part of a single population, as demonstrated by their close genetic affinities in multiple analyses (Supplementary Note 2; Supplementary Figures 1-10) and direct kinship connections (Figure 3-4). Our results indicate that individuals from the WBC sites and Sorsum predominantly chose partners from within this single population rather than integrating people from the outside, thus forming a cohesive reproductive unit and maintaining strict genetic boundaries, likely through social and/or cultural norms. One possible explanation for their close connection is that Sorsum was the northern most location of a WBC site, adopting some TRB material culture while staying in contact with the WBC populations. Alternatively, TRB-West and WBC may have had a common origin, with Sorsum preserving genetic continuity and exchange with WBC over time, despite cultural differences.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNo close ties between WBC/Sorsum and other megalithic groups\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWBC/Sorsum exhibit their closest external relationship (60-75 cM, likely above fourth degree) with samples from the Blätterhöhle cave in northwestern Germany, who in turn are connected to outliers (characterized by high WHG ancestry) from Mont Aimé in the Paris Basin (Figures 2,4; Supplementary Table 4). The large number of IBD segments and Y chromosome haplogroups shared between the individuals implies that their high WHG ancestry may be due to a common genetic origin (i.e., they had the same WHG ancestors).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSimilarly, when considering additional published contemporaneous megalithic groups, we detected two other large clusters of individuals sharing IBD segments (Supplementary Figure 12). These two clusters are in England/Scotland and Sweden, respectively, and do not show connections to each other or to our Sorsum/WBC network. These findings indicate that while megalithic societies may have shared architectural styles and certain cultural practices, genetic exchange through direct kinship did not extend across vast distances or across the English Channel and the Baltic Sea. Instead, kinship networks appear to have been maintained within more localized or regionally structured populations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIntra-site kinship relationships\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWithin the six communities analysed here, we identified 123 closely related pairs, comprising 44 first-degree and 79 second-degree relationships (Figure 3; Supplementary Table 3). In addition, we detected 39 third-degree related pairs (Supplementary Figure 13; Supplementary Table 3). About half (54%) of the 203 individuals have a close relative (first- or second-degree). This proportion is comparatively low when considering other megalithic populations\u003csup\u003e2,3,5\u003c/sup\u003e. In total, we identified 19 clusters of close relatives. Most clusters are relatively small, ranging from isolated pairs to groups of up to six individuals. Larger family networks are seen in Niedertiefenbach (12 individuals), Sorsum (two clusters with 11 and 20 individuals) and Warburg (13 individuals). In the Warburg necropolis, the family ties span two of the three burial complexes (Warburg I and Warburg III)\u003csup\u003e18\u003c/sup\u003e—indicating that they were used contemporaneously. The presence of single pairs or small clusters of individuals who are not closely related to the four big families could indicate that burial in the chambers was not restricted to them. Instead, WBC and TRB-West may have emphasized broader community membership, social affiliation or other forms of non-kin inclusion in their funerary practices.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePatrilineal inheritance without constraints on female mobility\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo investigate sex-specific mobility patterns within WBC and Sorsum, we examined pairwise mismatches between same-sex pairs (male-male and female-female). Our results reveal no significant differences in relatedness between male and female pairs (Supplementary Figure 14), suggesting a relatively egalitarian mobility for the two sexes. This pattern contrasts with studies on other megalithic groups, such as those from the TRB-North or the British and Irish Neolithic, which show evidence for patrilocality with exogamous marriage practices\u003csup\u003e2,3\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eInterestingly, despite this lack of strict patrilocality in WBC and Sorsum, we observed dominant paternal lineages within each community. The Y chromosome haplogroup I2c is prevalent in Niedertiefenbach, Altendorf and Sorsum, whereas I2a is most common in Warburg (Figures 2-3; Supplementary Table 1). This pattern suggests that the men involved in founding the different communities had distinct I lineages, which remained predominant over time. However, rather than suggesting dynastic rule or rigid male inheritance structures, it may reflect communal burial traditions where individuals were interred based on shared ancestry or social affiliation and not hierarchical status. Notably, females are underrepresented in all WBC and Sorsum sites, comprising only about 40% of all individuals (Supplementary Figure 15). Females might have been buried in different locations due to social status or ritual customs. A similar underrepresentation of females has also been observed in other megalithic grave communities with strong patrilinear family lineages\u003csup\u003e2–4\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eOur findings reveal a complex interaction pattern in central Europe during the Late Neolithic. The presence of first- and second-degree relationships spanning several sites, especially the father-son pair bridging Niedertiefenbach (WBC) with Sorsum (TRB-West), indicates that there was substantial contact between megalithic communities that were not only far apart (over 200 km), but also had cultural differences in ceramic styles. Their close relation is further substantiated by\u0026nbsp;the archaeological record which shows similarities in burial construction (rock-cut chamber of Sorsum and WBC gallery graves)\u003csup\u003e19\u003c/sup\u003e and artefacts (hunting equipment—such as transverse and triangular arrowheads and bone points)\u003csup\u003e20–22\u003c/sup\u003e. Collectively our evidence points towards a much higher mobility than previously thought\u003csup\u003e2,23\u003c/sup\u003e. Despite this high mobility, WBC and Sorsum lacked close kinship ties with other megalithic populations, revealing restricted mating practices. These observations add an important layer to our understanding of megalithic societies. Although the use of large blocks and collective burial practices is a common feature across regions with megalithic structures, our findings also reveal notable differences, as reflected in the varying spatial ranges of kinship ties among the buried individuals. These differences imply that the same social sphere may have held distinct meanings for the various groups. In conclusion, our results point not only to a spatial dimension of kinship relations in the Neolithic but also to the complex nature of the megalithic phenomenon.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eInclusion \u0026amp; ethics statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe followed the guidelines for ethical ancient DNA (aDNA) research on human remains\u003csup\u003e24\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSampling, DNA extraction, library preparation and whole genome sequencing\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe sampling and DNA extraction procedures were carried out in a specialized ancient DNA facility at Kiel University, as outlined previously\u003csup\u003e25\u003c/sup\u003e. We collected a total of 479 commingled skeletal elements from four Late Neolithic collective burial sites located in present-day Germany: Altendorf, Sorsum, Warburg and Z\u0026uuml;schen. Some individuals are represented by more than one skeletal element. The DNA was extracted from bone fragments (specific bone source for each sample that entered our analysis is detailed in Supplementary Table 1) and converted into partial uracil-DNA glycosylase (UDG) double-stranded libraries\u003csup\u003e26\u003c/sup\u003e. The libraries were submitted for whole genome shotgun sequencing on the Illumina HiSeq 6000 (2x100) platform at the Institute of Clinical Molecular Biology in Kiel.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRaw read processing and ancient DNA authentication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAdapter sequences were trimmed and paired-end reads were merged using ClipAndMerge version 1.7.7\u003csup\u003e27\u003c/sup\u003e. The resulting reads were then aligned to the human reference genome (build hg19) using BWA version 0.7.15\u003csup\u003e28\u003c/sup\u003e, with relaxed mapping stringency (flag -n 0.01) to account for the expected mismatches in aDNA. If published and newly generated datasets were available for a given individual, they were merged into a single BAM file with Samtools v1.12\u003csup\u003e29\u003c/sup\u003e. Duplicate reads were removed using DeDup version 0.12.1\u003csup\u003e27\u003c/sup\u003e. All samples were evaluated for deamination with DamageProfiler version 1.1\u003csup\u003e30\u003c/sup\u003e. Additionally, contamination was estimated based on mitochondrial DNA (mtDNA) using Schmutzi version 1.5.5.5\u003csup\u003e31\u003c/sup\u003e and the X chromosome in male samples using ANGSD version 0.935\u003csup\u003e32\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGeneration of pseudo-haploid genotype datasets\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe two terminal positions of reads in the BAM files were trimmed using bamUtil version 1.0.15\u003csup\u003e33\u003c/sup\u003e to decrease the impact of aDNA deamination prior to genotyping. Next, we used SequenceTools (https://github.com/stschiff/sequenceTools) version 1.2.2 to create pseudo-haploid genotypes at positions from the 1240K and Human Origins (HO) panels. Samples with fewer than 20,000 SNPs covered in the 1240K panel were removed from population genetic analyses. The filtered genotype dataset comprising 129 samples was integrated with the Allen Ancient DNA Resource (AADR) reference panels (version 62.0 of the 1240K and HO panels) \u0026ndash; a comprehensive collection of published aDNA data\u003csup\u003e34\u003c/sup\u003e. In addition, we incorporated previously published data from Altendorf (n\u0026nbsp;=\u0026nbsp;15) and Warburg (n\u0026nbsp;=\u0026nbsp;17), along with samples from Niedertiefenbach (n\u0026nbsp;=\u0026nbsp;40) and Rimbeck (n\u0026nbsp;=\u0026nbsp;2)\u003csup\u003e11,12\u003c/sup\u003e. This brought the total number of individuals associated with the WBC and Sorsum communities analysed in this study to 203 (Table 1).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cstrong\u003eInference of genetic sex and uniparental haplogroups\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe genetic sex of the samples was determined using two methods that assess the ratio of sequences aligning to sex chromosomes relative to autosomes\u003csup\u003e35,36\u003c/sup\u003e. The mtDNA haplogroup of each sample was inferred with HaploGrep2 version 2.4.0\u003csup\u003e37\u003c/sup\u003e. For male individuals, Y chromosome haplogroups were determined using yHaplo v1.1.2\u003csup\u003e38\u003c/sup\u003e, based on either the full set of ISOGG SNPs or a filtered set restricted to transversions only. The haplogroup assignment with the highest resolution was retained for each individual. For the genetic sex determination and uniparental haplogroup inference, a mapping and base quality threshold of 30 was applied.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAncestry inference: PCA, f-statistics and admixture modelling\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA principal component analysis (PCA) was performed on the merged HO dataset with smartpca version 16000\u003csup\u003e39\u003c/sup\u003e using the option \u0026ldquo;lsqproject: YES\u0026rdquo;. The ancient samples were projected onto the genetic variation of modern West Eurasian populations from the HO panel. We computed f4 statistics with qpDstat version 970\u003csup\u003e40\u003c/sup\u003e with the option \u0026ldquo;f4mode: YES\u0026rdquo;. In the f4 analyses (results described inSupplementary Note 2), the 1240K dataset was used, and the Mbuti was included as the outgroup. We conducted a hypothesis-free ancestry deconvolution analysis using ADMIXTURE version 1.3.0\u003csup\u003e41\u003c/sup\u003e in unsupervised mode, exploring 3 to 12 components with 200 bootstraps (Supplementary Note 2). The analysis was performed on a pruned HO dataset using PLINK version 1.90b6.21 with the command \u0026ldquo;--indep-pairwise 200 25 0.4\u0026rdquo;\u003csup\u003e42\u003c/sup\u003e. Hypothesis-driven admixture modelling with two sources was performed with qpAdm version 1201\u003csup\u003e40\u003c/sup\u003e using the option \u0026ldquo;allsnps: NO\u0026rdquo; (Supplementary Note 2). The set of sources and outgroups used for qpAdm modelling is available in the Supplementary Table 2header. Additionally, we used DATES version 753\u003csup\u003e43\u003c/sup\u003e to model the most recent time of admixture, considering a single pulse that led to the formation of Sorsum and the WBC communities (Supplementary Note 2). The following parameters were used in DATES: \u0026apos;binsize: 0.001\u0026rsquo;, \u0026apos;maxdis: 1.0\u0026rsquo;, \u0026apos;seed: 77\u0026rsquo;, \u0026apos;jackknife: YES\u0026rsquo;, \u0026apos;qbin: 10\u0026rsquo;, \u0026apos;runfit: YES\u0026rsquo;, \u0026apos;afffit: YES\u0026rsquo;, \u0026apos;lovalfit: 0.45\u0026rsquo;, \u0026apos;minparentcount: 1\u0026rsquo;.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAssessing sex-biased admixture\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe assessed the possibility of sex-biased admixture by estimating the ancestry proportions on autosomes and the X chromosome (results in Supplementary Note 2). To this end, we relied on the 1240K SNP panel, which we initially pruned using PLINK version 1.90b6.21 (command \u0026ldquo;--indep-pairwise 200 25 0.4\u0026rdquo;)\u003csup\u003e42\u003c/sup\u003e. After linkage disequilibrium pruning, 569,582 autosomal and X chromosomal 6,737 SNPs remained that were subsequently used for supervised ADMIXTURE modelling (version 1.3.0)\u003csup\u003e41\u003c/sup\u003e. Only samples with sufficient coverage for at least 1000 X chromosomal SNPs were included in the analysis. Due to the small number of variants available on the X chromosome, we resampled the autosomal SNPs 1000 times and matched the number of X chromosomal SNPs. For each individual, we calculated the ratio of WHG ancestry on the X chromosomes to the WHG ancestry on autosomes (R\u003csub\u003eX/A\u003c/sub\u003e) and performed a log\u003csub\u003e2\u003c/sub\u003e transformation of this statistic to better visualize deviations from the null hypothesis (i.e., no imbalance between the ancestral proportions of autosomes and X chromosomes). \u0026nbsp;A Wilcoxon signed-rank test was performed to determine whether the differences in estimated WHG ancestry between autosomes and X chromosomes were statistically significant\u003csup\u003e44\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKinship analysis for Sorsum and the WBC communities\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo assess kinship among individuals from Sorsum and the WBC sites, we used the following three programs: READv2, KIN, and NGSRelate2\u003csup\u003e14\u0026ndash;16\u003c/sup\u003e. As we observed a difference in the baseline mismatch rate between pairs from Sorsum (0.274) and those from WBC populations (0.252) (Supplementary Figure 16), which may lead to under- or overestimation of relatedness, we conducted separate analyses to ensure accurate kinship assessment. First, we only considered Sorsum samples to identify kinship within this population. Second, we only analysed WBC samples to assess kinship exclusively within this group. Finally, we combined samples from both Sorsum and WBC sites to identify kinship between the two populations. The analyses were conducted using SNPs from the 1240K panel, applying the following filters: kinship estimates were disregarded for a pair if fewer than 2,000 shared SNPs were available (for READv2 and NGSRelate2) or if the log-likelihood ratio was below 1 (for KIN). All three methods yielded highly consistent results, attesting to the reliability of the estimates (Supplementary Figure 17). To conservatively construct the close kinship network, we used a majority-rule consensus approach, where kinship assignments were determined based on agreement from at least two of the three methods. For example, if one program classified a pair as first-degree relatives while the other two assigned them as second-degree, we considered them second-degree kin. Pairs for which all three methods produced conflicting results were excluded from the kinship network. To ensure robustness, we primarily focused on first- and second-degree relationships as kinship estimates beyond this range carry greater uncertainty. For completeness, third-degree kinship estimates were also included in the Supplementary Information (Supplementary Figure 13; Supplementary Table 3).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentifying IBD segments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe employed ancIBD version 0.3a1\u003csup\u003e17\u003c/sup\u003e to identify identity-by-descent (IBD) segments. These segments were primarily utilized to explore more distant relationships between individuals from Sorsum, WBC and other contemporaneous groups in the region (4000 \u0026ndash; 2000 BCE; Supplementary Table 4). Additionally, the ancIBD results were also used to validate the kinship established through the majority-rule consensus method outlined above, whenever possible. For this analysis, we first imputed data from high-coverage ancient genomes (mean depth \u0026ge; 0.1X) using GLIMPSE version 2.0.0\u003csup\u003e45\u003c/sup\u003e. Imputation was done with the 1000 Genomes Project reference panel. ancIBD was run with the default parameters. Since most biological relatives up to the sixth degree share at least two long IBD segments\u003csup\u003e46\u003c/sup\u003e, we included only pairs with three or more IBD segments larger than 12 cM in the IBD networks. Pedsim version 1.4\u003csup\u003e46\u003c/sup\u003e was used to simulate IBD segments across several kinship scenarios for a comparison against the IBD estimates observed in the ancient populations.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEstimating ROH and N\u003csub\u003ee\u003c/sub\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe analysed runs of homozygosity (ROH) and effective population size (Ne) using HapROH version 0.3a4\u003csup\u003e47\u003c/sup\u003e on high-coverage samples (\u0026ge; 400,000 SNPs from the 1240K panel covered) from Sorsum, WBC and other published groups between 4000 and 2000 BCE (results described in Supplementary Note 2). The program was executed with default parameters.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the sequenced data generated in this study is deposited to the European Nucleotide Archive (accession: PRJEB88326).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) through the CRC 1266 – project number 290391021 and the Clusters of Excellence ROOTS (EXC 2150 ID 390870439).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eB.K.-K. and J.Mü. designed the research.\u0026nbsp;F.K., I.G., K.S. and M.R.\u0026nbsp;provided the archaeological materials and context information. Laboratory work was supervised by B.K.-K. N.A.d.S. primarily performed the analyses, with assistance from D.K. and D.A.M. Ch.R., J.Me. and J.Mü. contributed to embedding the findings into the broader chronological and archaeological framework.\u0026nbsp;K.F. and Ch.M. curated the samples.\u0026nbsp;N.A.d.S., A.N., J.Mü. and B.K.-K. interpreted the results and wrote the manuscript, with all the authors reviewing. A.N. and B.K.-K. secured funding and supervised the project.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaterials \u0026amp; Correspondence\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCorrespondence to Ben Krause-Kyora ([email protected])\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSchulz Paulsson, B. \u003cem\u003eTime and Stone : The Emergence and Development of Megaliths and Megalithic Societies in Europe\u003c/em\u003e. (Archaeopress Publishing, Oxford, 2017).\u003c/li\u003e\n\u003cli\u003eSeersholm, F. 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Accurate sex identification of ancient human remains using DNA shotgun sequencing. \u003cem\u003eJ. Archaeol. Sci.\u003c/em\u003e \u003cstrong\u003e40\u003c/strong\u003e, 4477\u0026ndash;4482 (2013).\u003c/li\u003e\n\u003cli\u003eWeissensteiner, H. \u003cem\u003eet al.\u003c/em\u003e HaploGrep 2: mitochondrial haplogroup classification in the era of high-throughput sequencing. \u003cem\u003eNucleic Acids Res.\u003c/em\u003e \u003cstrong\u003e44\u003c/strong\u003e, W58-63 (2016).\u003c/li\u003e\n\u003cli\u003ePoznik, G. D. Identifying Y-chromosome haplogroups in arbitrarily large samples of sequenced or genotyped men. 088716 Preprint at https://doi.org/10.1101/088716 (2016).\u003c/li\u003e\n\u003cli\u003ePatterson, N., Price, A. L. \u0026amp; Reich, D. 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M. \u003cem\u003eet al.\u003c/em\u003e The formation of human populations in South and Central Asia. \u003cem\u003eScience\u003c/em\u003e \u003cstrong\u003e365\u003c/strong\u003e, eaat7487 (2019).\u003c/li\u003e\n\u003cli\u003eGoldberg, A., G\u0026uuml;nther, T., Rosenberg, N. A. \u0026amp; Jakobsson, M. Ancient X chromosomes reveal contrasting sex bias in Neolithic and Bronze Age Eurasian migrations. \u003cem\u003eProc. Natl. Acad. Sci.\u003c/em\u003e \u003cstrong\u003e114\u003c/strong\u003e, 2657\u0026ndash;2662 (2017).\u003c/li\u003e\n\u003cli\u003eRubinacci, S., Hofmeister, R. J., Sousa da Mota, B. \u0026amp; Delaneau, O. Imputation of low-coverage sequencing data from 150,119 UK Biobank genomes. \u003cem\u003eNat. Genet.\u003c/em\u003e \u003cstrong\u003e55\u003c/strong\u003e, 1088\u0026ndash;1090 (2023).\u003c/li\u003e\n\u003cli\u003eCaballero, M. \u003cem\u003eet al.\u003c/em\u003e Crossover interference and sex-specific genetic maps shape identical by descent sharing in close relatives. \u003cem\u003ePLOS Genet.\u003c/em\u003e \u003cstrong\u003e15\u003c/strong\u003e, e1007979 (2019).\u003c/li\u003e\n\u003cli\u003eRingbauer, H., Novembre, J. \u0026amp; Steinr\u0026uuml;cken, M. Parental relatedness through time revealed by runs of homozygosity in ancient DNA. \u003cem\u003eNat. Commun.\u003c/em\u003e \u003cstrong\u003e12\u003c/strong\u003e, 5425 (2021).\u003c/li\u003e\n\u003cli\u003eRinne, C., Drummer, C. \u0026amp; Hamann, C. Collective and individual burial practices. Changing patterns at the beginning of the third millennium BC: The megalithic grave of Altendorf. \u003cem\u003eJ. Neolit. Archaeol.\u003c/em\u003e 75\u0026ndash;88 (2019) doi:10.12766/jna.2019.4.\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003e\u003cbr /\u003e \u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1.\u0026nbsp;\u003c/strong\u003eOverview of archaeological sites analysed in this study. N: number of individuals with whole genome sequencing data.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eArchaeological site\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eDating (BCE)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eN (previously published)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eN (newly generated)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eN total\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAltendorf\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3250 – 3100\u003csup\u003e48\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eNiedertiefenbach\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3350 – 3250\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eRimbeck\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3350 – 3075\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eSorsum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3350 – 3100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eWarburg\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3350 – 2950\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e71\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eZüschen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3500 – 3150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003e74\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003e129\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003e203\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e* Data provided as personal communication from C. Rinne.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-6464608/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6464608/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Across Neolithic Europe, megalithic monuments – including stone circles and gallery graves – were constructed by early farming communities, representing a supra-regional cultural phenomenon1. The collective graves are often viewed as symbols of shared ancestry, social identity and regional connectivity. Palaeogenetic studies of megalithic burial sites have identified patrilineal kinship and locally restricted female mobility (≤ 8 km)2–5. However, until now, close genetic ties between geographically distant populations have not been described. Here we analyse genome-wide data from 203 individuals associated with five Wartberg and one Western Funnel Beaker communities. Our results reveal long-distance first- and second-degree kinship ties across these megalithic sites, involving both females and males. Remarkably, we identify close relatives — such as a father and son — who were buried more than 200 kilometres apart and belonged to distinct cultures. Additionally, we uncover extended genetic networks that link the six communities. Together, they form a single population with genetic boundaries to other megalithic societies. Our findings suggest that mobility and mating networks spanned hundreds of kilometres, fostering strong intra-group cohesion while maintaining limited external contacts. This indicates that the supra-regional megalithic phenomenon may not reflect deep social integration, but rather a shared cultural expression without strong underlying biological connections.","manuscriptTitle":"Long-distance kinship in megalithic Europe","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-07 04:52:35","doi":"10.21203/rs.3.rs-6464608/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"acc58e9e-ed7d-41e1-bd8c-1d4c8864a9ee","owner":[],"postedDate":"May 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":48099941,"name":"Biological sciences/Genetics/Genomics/Genome evolution"},{"id":48099942,"name":"Biological sciences/Evolution/Archaeology"},{"id":48099943,"name":"Biological sciences/Evolution/Anthropology/Biological anthropology"},{"id":48099944,"name":"Biological sciences/Genetics/Population genetics"}],"tags":[],"updatedAt":"2025-06-08T13:41:24+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-07 04:52:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6464608","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6464608","identity":"rs-6464608","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}