Statistical Methods for Identifying Demographic Structure in DNA Sequence Alignments

All life on Earth, from viruses and bacteria, trees and flowers, to birds and human beings, can be traced back to a single common ancestor. However, the evolutionary history that led to this diversity of life is a complicated story that we do not yet fully understand. Since the discovery of the structure of deoxyribonucleic acid (DNA) in 1953, and the development of DNA sequencing technology, researchers have been using similarities and differences in the genomes of organisms to better understand the relationships between species. However, due to the complexity of the evolutionary history of life, simplifying assumptions must be made to make mathematical models tractable. It must then be of paramount importance for researchers to be able to identify when the simplifying assumptions of a specific model are unreasonable. In this thesis we present two projects, and although they are different in implementation, both attempt to investigate simplifying assumptions in the closely related fields of population genetics and phylogenetics. However, we also present applications of our projects where the results of our work are not used in assessing assumptions for further analyses, but are of standalone interest to researchers. Our first project is concerned with the development of a method for constructing coordinate representations for single-copy DNA, such as mitochondrial DNA (mtDNA) or Y-chromosomal DNA, analogous to the use of PCA for nuclear DNA. We construct a coordinate system such that, given p informative sites in an alignment of n individuals, returns p-dimensional coordinates for each n individuals. We order the dimensions by the proportion of variability each dimension captures in the overall genetic diversity. From these coordinates in \genetic space" researchers may perform a number of down stream analyses. It is possible to optimally visualise high-dimensional sequence data in two or three dimensions. One may use our method to identify closely related individuals, identify sites in the alignment that are closely linked, or to use the same coordinate space to nd sites that are closely linked with groups of individuals. Finally, one may choose to test for significant relationships between the structure of the coordinates in genetic space, and metadata recorded on sequenced individuals, indicating demographic variables that are highly related to the evolutionary history of an alignment. This final application of our method, where one may test for demographic structure in sequence data, is of key importance to the theme of discovering when simplifying assumptions of analyses are not reasonable. Through the comparison of coordinates in gene space, and any demographic variables of interest, researchers may explore whether or not the individuals in the alignment indicate population substructure. For example, one may investigate if there appears to be a phylogeographic structure to the individuals forming distinct subpopulations, and if migration appears to occur between subpopulations. Through empirical data, we show that our method can readily recover tree-like structure, identify strong genetic groupings based on qualitative traits and show that we are able to recover phylogeographic signal given provenanced sampling information. We show that our method can even be used to suggest routes of migration based on mtDNA. Finally we apply our method to modern Aboriginal Australian mtDNA to show strong evidence for discrete geographic populations of Aboriginal Australian peoples that display permanence on the Australian landscape dating back to the original colonisation of Australia 50 thousand years before present (kya). Our second project is concerned with identifying departures from a tree-like evolutionary history at the species level. It is not uncommon for closely related species (Species A and C say) to still be capable of interbreeding, and producing viable \hybrid" offγspring (Species B say). Under these conditions, a phylogenetic tree cannot describe the evolutionary history of the hybrid species, and instead an admixture graph may be a better description. We begin by considering the evolutionary history of three species: a hybrid organism that has undergone some independent evolution (Species B), and two \parent" organisms, Species A and C. Relatively long, contiguous regions of the genome of Species B will have undergone no recombination since the admixture event. These regions will have been contributed by either Species A (and hence will be more closely related to Species A), or Species C. We aim to estimate the proportion of the genome contributed by Species A, and denote this by considering the proportion of informative site patterns that indicate evidence for the two possible ancestries. The mixing proportion is the parameter of interest in our analyses. However, due to the classical problem of the non-identifiability of mixing parameters in multinomial distributions, we describe two Bayesian methods for estimating γ. Our first method places prior distributions on the parameters of the model, and uses Approximate Bayesian Computation (ABC) to estimate the marginal posterior distribution of γ. Our second, closely related method, instead estimates the marginal posterior distribution of via numerical integration. We show via a simulation study that our methods can accurately estimate the true value of γ, and perform well under biologically reasonable scenarios. However, we also find that our methods suffer from a relatively small positive bias for small values of γ, i.e., when one species of the parent species contributes very little to the genome of the hybrid species. We compare the performance of our method to the popular method of the ratio of f4 statistics. We do this by estimating the proportion of Neanderthal ancestry in pre-ice age European human samples and comparing our results to the finding of Fu et al. [18]. We show that our method recovers extremely similar estimates of Neanderthal ancestry with no apparent systematic bias when compared to the results of Fu et al.. Finally we apply our method to the genomes of Late Pleistocene European bison (Bison bonasus) and Steppe Bison (Bison priscus) to understand the evolutionary history of bovid megafauna in Europe over the last seventy thousand years. It was thought that before 10 kya the only bovid present in Europe was the Steppe bison. However, from bone samples found dating from the present day, and back to approximately 70 kya, mtDNA indicated a second bison species was also roaming Europe before 10 kya, more closely related to modern cattle than the Steppe bison. After nuclear DNA was sequenced, we were able to show that this new species of bovid was actually a hybrid offspring of Aurochs (the ancestor of modern cattle) and Steppe bison, an event that occurred approximately 120 kya. We used our method, in concert with the ratio of f4 statistics, to show that the hybrid species contained approximately 10% Aurochs and 90% Steppe bison ancestry.Thesis (Ph.D.) -- University of Adelaide, School of Mathematical Sciences, 201

A Woman with a Sword? : Weapon Grave at Suontaka Vesitorninmäki, Finland

Publisher Copyright: © 2021 European Association of Archaeologists.In 1968, a weapon grave with brooches was found at Suontaka Vesitorninmäki, Hattula, Finland. Since then, the grave has been interpreted as evidence of powerful women, even female warriors and leaders in early medieval Finland. Others have denied the possibility of a woman buried with a sword and tried to explain it as a double burial. We present the first modern analysis of the grave, including an examination of its context, a soil sample analysis for microremains, and an aDNA analysis. Based on these analyses, we suggest a new interpretation: the Suontaka grave possibly belonged to an individual with sex-chromosomal aneuploidy XXY. The overall context of the grave indicates that it was a respected person whose gender identity may well have been non-binary.Peer reviewe

Intramural child burials in Iron Age Navarra: How ancient DNA can contribute to household archaeology

The transition from the Late Bronze to the Iron Age on the Iberian Peninsula saw a shift in mortuary customs from mainly inhumation to cremation of the deceased. The poor preservation characteristic of cremated skeletal remains has hindered molecular analyses (isotope analyses, ancient DNA) of the Iberian Final Bronze and Iron Age communities of Iberia. Incidentally, a limited number of young children, often newborns, were exempt from the predominant cremation ritual, in favour of intramural inhumations inside buildings at certain settlements. The discourse surrounding the mean- ing and interpretation of this particular burial rite has developed over a long time in Iberian archaeology but has always been hampered by the limited anthropological, archaeological, and molecular data from these intramural inhumations. Here, we study the genomes of 37 intramurally buried children found in three Early Iron Age settlements, dated between c. 800–450 BC. Population genetic analyses on the newly reported individuals extend our understanding of ancient Iberia by revealing previously unsampled genetic diversity as well as showing a lesser influence of Mediterranean ancestry than on previously published Iron Age individuals from northern Spain. We also provide insights into the sex and biological relatedness of the children, and in so doing, elucidate differ- ent aspects of the intramural burial ritual and building use in settlements. More broadly, the genetic data from these individuals fill an important gap in the archaeogenetic record of northern Spain and offer a unique opportunity to study the genetic makeup and population changes from the Bronze Age to Antiquity.This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement number 851511). It has also been supported by the research project »Convergence and interaction between complex Bronze Age societies« from the Academia program of the Institució Catalana de Recerca i Estudis Avançats (ICREA) of the Catalan Government and the Spanish Ministry for Science and Innovation (PID2020-112909GB-100)

A Woman with a Sword? – Weapon Grave at Suontaka Vesitorninmäki, Finland

In 1968, a weapon grave with brooches was found at Suontaka Vesitorninmäki, Hattula, Finland. Since then, the grave has been interpreted as evidence of powerful women, even female warriors and leaders in early medieval Finland. Others have denied the possibility of a woman buried with a sword and tried to explain it as a double burial. We present the first modern analysis of the grave, including an examination of its context, a soil sample analysis for microremains, and an aDNA analysis. Based on these analyses, we suggest a new interpretation: the Suontaka grave possibly belonged to an individual with sex-chromosomal aneuploidy XXY. The overall context of the grave indicates that it was a respected person whose gender identity may well have been non-binary.</p

A 1000-year-old case of Klinefelter's syndrome diagnosed by integrating morphology, osteology, and genetics

We thank the Municipality of Bragança, the University of Coimbra, the University of Adelaide, the Max Planck Society, and the Calouste Gulbenkian Foundation for the support provided. ST is supported by the Fundação para a Ciência e a Tecnologia (SFRH/BD/116363/2016). BL (FT170100448) and JCT (DE210101235) are supported by the Australian Research Council. ABR is supported by the European Research Council (771234-PALEoRIDER)

Genomic history of Neolithic to Bronze Age Anatolia, Northern Levant, and Southern Caucasus

Here, we report genome-wide data analyses from 110 ancient Near Eastern individuals spanning the Late Neolithic to Late Bronze Age, a period characterized by intense interregional interactions for the Near East. We find that 6th millennium BCE populations of North/Central Anatolia and the Southern Caucasus shared mixed ancestry on a genetic cline that formed during the Neolithic between Western Anatolia and regions in today’s Southern Caucasus/Zagros. During the Late Chalcolithic and/or the Early Bronze Age, more than half of the Northern Levantine gene pool was replaced, while in the rest of Anatolia and the Southern Caucasus, we document genetic continuity with only transient gene flow. Additionally, we reveal a genetically distinct individual within the Late Bronze Age Northern Levant. Overall, our study uncovers multiple scales of population dynamics through time, from extensive admixture during the Neolithic period to long-distance mobility within the globalized societies of the Late Bronze Age. Video Abstrac

Cases of trisomy 21 and trisomy 18 among historic and prehistoric individuals discovered from ancient DNA

Aneuploidies, and in particular, trisomies represent the most common genetic aberrations observed in human genetics today. To explore the presence of trisomies in historic and prehistoric populations we screen nearly 10,000 ancient human individuals for the presence of three copies of any of the target autosomes. We find clear genetic evidence for six cases of trisomy 21 (Down syndrome) and one case of trisomy 18 (Edwards syndrome), and all cases are present in infant or perinatal burials. We perform comparative osteological examinations of the skeletal remains and find overlapping skeletal markers, many of which are consistent with these syndromes. Interestingly, three cases of trisomy 21, and the case of trisomy 18 were detected in two contemporaneous sites in early Iron Age Spain (800-400 BCE), potentially suggesting a higher frequency of burials of trisomy carriers in those societies. Notably, the care with which the burials were conducted, and the items found with these individuals indicate that ancient societies likely acknowledged these individuals with trisomy 18 and 21 as members of their communities, from the perspective of burial practice

Dynamic changes in genomic and social structures in third millennium BCE central Europe

Europe’s prehistory oversaw dynamic and complex interactions of diverse societies, hitherto unexplored at detailed regional scales. Studying 271 human genomes dated ~4900 to 1600 BCE from the European heartland, Bohemia, we reveal unprecedented genetic changes and social processes. Major migrations preceded the arrival of “steppe” ancestry, and at ~2800 BCE, three genetically and culturally differentiated groups coexisted. Corded Ware appeared by 2900 BCE, were initially genetically diverse, did not derive all steppe ancestry from known Yamnaya, and assimilated females of diverse backgrounds. Both Corded Ware and Bell Beaker groups underwent dynamic changes, involving sharp reductions and complete replacements of Y-chromosomal diversity at ~2600 and ~2400 BCE, respectively, the latter accompanied by increased Neolithic-like ancestry. The Bronze Age saw new social organization emerge amid a ≥40% population turnover.Peer reviewe

Genomic transformation and social organization during the Copper Age-Bronze Age transition in southern Iberia

The emerging Bronze Age (BA) of southeastern Iberia saw marked social changes. Late Copper Age (CA) settlements were abandoned in favor of hilltop sites, and collective graves were largely replaced by single or double burials with often distinctive grave goods indirectly reflecting a hierarchical social organization, as exemplified by the BA El Argar group. We explored this transition from a genomic viewpoint by tripling the amount of data available for this period. Concomitant with the rise of El Argar starting ∼2200 cal BCE, we observe a complete turnover of Y-chromosome lineages along with the arrival of steppe-related ancestry. This pattern is consistent with a founder effect in male lineages, supported by our finding that males shared more relatives at sites than females. However, simple two-source models do not find support in some El Argar groups, suggesting additional genetic contributions from the Mediterranean that could predate the BA.This work was supported by the Max Planck Society (V.V.-M. and W.H.); European Research Council (ERC) grant 771234—PALEoRIDER (W.H.); Spanish Ministry of Economy, Industry and Competitiveness project HAR2017-85962-P (C.O., C.R.-H., M.I.F., E.C.B., C.V.-F., V.L., R.M., and R.R.); AGAUR 2017SGR1044 (C.O., C.R.-H., M.I.F., E.C.B., C.V.-F., V.L., R.M., and R.R.); ICREA Academia program (R.R.); John Templeton Foundation grant 61220 (D.R.); and Paul Allen Family Foundation (D.R.). D.R. is an Investigator of the Howard Hughes Medical Institute

Genomic transformation and social organization during the Copper Age-Bronze Age transition in southern Iberia

[EN]The emerging Bronze Age (BA) of southeastern Iberia saw marked social changes. Late Copper Age (CA) settlements were abandoned in favor of hilltop sites, and collective graves were largely replaced by single or double burials with often distinctive grave goods indirectly reflecting a hierarchical social organization, as exemplified by the BA El Argar group. We explored this transition from a genomic viewpoint by tripling the amount of data available for this period. Concomitant with the rise of El Argar starting similar to 2200 cal BCE, we observe a complete turnover of Y-chromosome lineages along with the arrival of steppe-related ancestry. This pattern is consistent with a founder effect in male lineages, supported by our finding that males shared more relatives at sites than females. However, simple two-source models do not find support in some El Argar groups, suggesting additional genetic contributions from the Mediterranean that could predate the BA.This work was supported by the Max Planck Society (V.V.-M. and W.H.); European Research Council (ERC) grant 771234-PALEoRIDER (W. H.); Spanish Ministry of Economy, Industry and Competitiveness project HAR2017-85962-P (C.O., C.R.-H., M.I.F., E.C.B., C.V.-F., V.L., R.M., and R.R.); AGAUR 2017SGR1044 (C.O., C.R.-H., M.I.F., E. C.B., C.V.-F., V. L., R.M., and R.R.); ICREA Academia program (R.R.); John Templeton Foundation grant 61220 (D.R.); and Paul Allen Family Foundation (D.R.). D.R. is an Investigator of the Howard Hughes Medical Institute