Genetic transitions in the Neolithic and Bronze Age at Mas d'en Boixos (Catalonia, Spain)

Xavier Roca-Rada^{1

2}, Daniel R Cuesta-Aguirre³, Diana C Vinueza-Espinosa³, Roberta Davidson¹, Shyamsundar Ravishankar¹, Leonard Taufik^{1

4

5}, Núria Armentano^{3

6}, Xavier Esteve⁷, Yassine Souilmi^{1

4

8

9}, João C Teixeira^{1

10

11

12}, Assumpció Malgosa³, Bastien Llamas^{1

4

8

9

10}, Cristina Santos³

Affiliations

¹ Australian Centre for Ancient DNA, School of Biological Sciences, University of Adelaide, Adelaide, SA, Australia.
² Faculty of Arts and Humanities, University of Coimbra, Coimbra, Portugal.
³ Unitat Antropologia Biològica and GREAB - Grup de Recerca en Antropologia Biològica, Universitat Autònoma de Barcelona, Barcelona, Spain.
⁴ Indigenous Genomics, The Kids Research Institute Australia, Adelaide, SA, Australia.
⁵ Mochtar Riady Institute for Nanotechnology, Tangerang, Indonesia.
⁶ Museu d'Arqueologia de Catalunya, Barcelona, Spain.
⁷ Servei de Patrimoni Arqueològic i Paleontològic-Generalitat de Catalunya, ERAAUB, IAUB, Barcelona, Spain.
⁸ Environment Institute, University of Adelaide, Adelaide, SA, Australia.
⁹ National Centre for Indigenous Genomics, Australian National University, Canberra, ACT, Australia.
¹⁰ Centre of Excellence for Australian Biodiversity and Heritage (CABAH), University of Adelaide, Adelaide, SA, Australia.
¹¹ Evolution of Cultural Diversity Initiative, Australian National University, Canberra, ACT, Australia.
¹² Centre for Interdisciplinary Studies, University of Coimbra, Coimbra, Portugal.

PMID: 40687794
PMCID: PMC12273517
DOI: 10.1016/j.isci.2025.112871

Genetic transitions in the Neolithic and Bronze Age at Mas d'en Boixos (Catalonia, Spain)

Xavier Roca-Rada et al. iScience. 2025.

. 2025 Jun 11;28(7):112871.

doi: 10.1016/j.isci.2025.112871. eCollection 2025 Jul 18.

Authors

Affiliations

¹ Australian Centre for Ancient DNA, School of Biological Sciences, University of Adelaide, Adelaide, SA, Australia.
² Faculty of Arts and Humanities, University of Coimbra, Coimbra, Portugal.
³ Unitat Antropologia Biològica and GREAB - Grup de Recerca en Antropologia Biològica, Universitat Autònoma de Barcelona, Barcelona, Spain.
⁴ Indigenous Genomics, The Kids Research Institute Australia, Adelaide, SA, Australia.
⁵ Mochtar Riady Institute for Nanotechnology, Tangerang, Indonesia.
⁶ Museu d'Arqueologia de Catalunya, Barcelona, Spain.
⁷ Servei de Patrimoni Arqueològic i Paleontològic-Generalitat de Catalunya, ERAAUB, IAUB, Barcelona, Spain.
⁸ Environment Institute, University of Adelaide, Adelaide, SA, Australia.
⁹ National Centre for Indigenous Genomics, Australian National University, Canberra, ACT, Australia.
¹⁰ Centre of Excellence for Australian Biodiversity and Heritage (CABAH), University of Adelaide, Adelaide, SA, Australia.
¹¹ Evolution of Cultural Diversity Initiative, Australian National University, Canberra, ACT, Australia.
¹² Centre for Interdisciplinary Studies, University of Coimbra, Coimbra, Portugal.

PMID: 40687794
PMCID: PMC12273517
DOI: 10.1016/j.isci.2025.112871

Abstract

Mas d'en Boixos is a key prehistoric site in Northeastern Iberia spanning from the Early Neolithic to the Late Iron Age. We analyzed genome-wide data from eight individuals and ten mitogenomes, dated to the Middle Neolithic and Early Bronze Age, alongside three previously published Iron Age individuals. Two Middle Neolithic individuals buried together were first-degree maternal relatives and carried Western Hunter-Gatherer, Anatolian, and residual Magdalenian-associated ancestries. Conversely, six Early Bronze Age individuals buried in a hypogeum exhibited distinct mitochondrial lineages. Among them, three were third-degree relatives, and all males shared a Y-chromosome lineage, consistent with a collective burial of an extended family within a patrilocal society practicing possible female exogamy. These individuals showed genetic continuity with additional Steppe-related ancestry, which displayed a subtle southward gradient across Iberia. We also identified an Eastern European mitochondrial lineage-challenging the proposed male-driven Bronze Age transition-and Mediterranean gene flow-suggesting dynamic interactions across the sea.

Keywords: Biological sciences; Human genetics; Paleobiology.

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Conflict of interest statement

The authors declare no competing interests.

Figures

**Figure 1**
Location and topography of Mas d’en Boixos in the Catalan pre-littoral depression of the Penedès region (Barcelona, Catalonia, Spain) and the positions of the archaeological structures discussed in this study

**Figure 2**
The Early Bronze Age hypogeum (E35) from MDB

**Figure 3**
Genetic structure and ancestry transitions in ancient Iberia and MDB (A) PCA of present-day Western Europeans (light gray) with projected ancient Iberians as colored circles (Upper Palaeolithic (UP) and Mesolithic (M) Iberian Hunter-Gatherers, N: Neolithic; C: Chalcolithic; BA: Bronze Age; and IA: Iron Age), including the MDB individuals as colored diamonds. (B) Iterations of outgroup f₃-statistics of the form f₃ (Mbuti; Ind1, Ind2) visualized in a multidimensional scaling analysis of 1 - f₃. (C) f₄-statistics are displayed as the mean ± 1-SD with gray bars representing Z scores ≤3 and black bars Z scores ≥3. Results show an increased affinity to Russia_Samara_EBA_Yamnaya (absent in MDB_MN) which implies the presence of Steppe-related ancestry in MDB_EBA and MDB_IA. This plot also shows the turnover in Y-chromosome lineages in male individuals (color-filled squares) and the origin of their mtDNA haplogroups.

**Figure 4**
Ancestry proportions estimated with a supervised ADMIXTURE at k = 4, with the four predefined ancestries being Western Hunter-Gatherers (WHG), Turkish individuals from the Neolithic representing the Anatolian Farmers ancestry (Anatolia_N), Bronze Age Eurasians from the Yamnaya culture representing the Steppe-related ancestry (Russia_Samara_EBA_Yamnaya) and present-day African individuals being proxy for Sub-Saharan ancestry (Mbuti) with ancient Iberians from the pre-Neolithic (hunter-gatherers; HG), Neolithic (N), Chalcolithic (C), Bronze Age (BA) and Iron Age (IA)

**Figure 5**
Steppe-related ancestry in ancient Iberia and MDB (A) Ancestry proportions for MDB temporal groups using *qpAdm*; p values are provided inside each column. (B) The Russia_Samara_EBA_Yamnaya ancestry proportion in Iberia during the Bronze Age computed with f₄-ratios colored from highest to lowest using a color scale. The MDB individuals are represented as triangles. The Russia_Samara_EBA_Yamnaya ancestry proportion decreases southwards. (C) Outgroup f₃-statistics of the form f₃(Mbuti; Pop, X) showing the amount of shared drift between the ancient individuals (X) with Turkish individuals from the Neolithic representing the Anatolian Farmers ancestry (Anatolia_N; x-axis) and Bronze Age Eurasians from the Yamnaya culture representing the Steppe ancestry (Russia_Samara_EBA_Yamnaya; y-axis). Clusters of individuals are represented in shades of blue, pink, and amber from the top-left to the bottom-right. The MDB individuals are represented as colored diamonds.

See this image and copyright information in PMC

References

1. Bouso M., Esteve X., Farré J., Feliu J.M., Palomo A., Rodríguez A., Senabre M.R., Mestres J. Del Bronze inicial a la depressió prelitoral catalana: Can Roqueta (Sabadell, Vallès Occidental) i Mas d’en Boixos-1 (Pacs del Penedès, Alt Penedès) CYPSELA. 2004;15:73–101.
1. Pascual M.P., Mercadé J.M. Vol. 2011. Jornades d’Arqueologia del Penedès; 2016. Mas d’en Boixos (Pacs del Penedès, Alt Penedès) pp. 137–148. (Resultats de la campanya d’intervenció arqueològica de l’any 2008).
1. Farré, J., Mestres, J., Senabre, M.R., and Feliu, J.M. El jaciment de Mas d’en Boixos (Pacs del Penedès, Alt Penedès). Un espai utilitzat des del Neolític fins a l’època ibèrica, Generalitat de Catalunya.
1. Broushaki F., Thomas M.G., Link V., López S., van Dorp L., Kirsanow K., Hofmanová Z., Diekmann Y., Cassidy L.M., Díez-del-Molino D., et al. Early Neolithic genomes from the eastern Fertile Crescent. Science. 2016;353:499–503. doi: 10.1126/science.aaf7943. - DOI - PMC - PubMed
1. Skoglund P., Malmström H., Raghavan M., Storå J., Hall P., Willerslev E., Gilbert M.T.P., Götherström A., Jakobsson M. Origins and Genetic Legacy of Neolithic Farmers and Hunter-Gatherers in Europe. Science. 2012;336:466–469. doi: 10.1126/science.1216304. - DOI - PubMed

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Genetic transitions in the Neolithic and Bronze Age at Mas d'en Boixos (Catalonia, Spain)

Affiliations

Genetic transitions in the Neolithic and Bronze Age at Mas d'en Boixos (Catalonia, Spain)

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

LinkOut - more resources

Full Text Sources

Research Materials