Kinship between burials from Grave Circle B at Mycenae revealed by ancient DNA typing

The richness of the burials in Grave Circle B at Mycenae, Greece indicates that the 35 people interred there held elite status during their lifetimes 3500 years ago. It has been speculated that the groups of burials represent different dynasties or branches of the same family. To test this hypothesis, we carried out an exhaustive ancient DNA (aDNA) study of 22 of the skeletons. We were unable to identify nuclear aDNA in any specimen, but we obtained authentic mitochondrial aDNA sequences for four individuals. The results were compared with facial reconstructions and interpreted within the archaeological context represented by the organisation of the graves and the positions of the burials within the graves. We conclude that the contemporaneous male Γ55 and female Γ58 skeletons, which both possess the UK mitochondrial haplogroup, were brother and sister. The implication is that Γ58 was buried in Grave Circle B not because of a marital connection but because she held a position of authority by right of birth. The results illustrate the difficulty in using aDNA to study kinship relationships between archaeological specimens, but also show that aDNA can advance understanding of kinship when used to test hypotheses constructed from other evidence.     

Ancient DNA analysis of desiccated wheat grains excavated from a Bronze Age cemetery in Xinjiang

Wheat has been one of the most important crop in Eurasia since the Neolithic period. Understanding the spread of wheat cultivation is crucial to understanding the spread of agriculture as a whole and the interactions between prehistoric populations across the Eurasian continent. However, the routes by which wheat cultivation spread eastwards have been poorly understood to date, due to the scarcity of plant remains recovered from archaeological sites. Desiccated wheat grains excavated from the Xiaohe cemetery in Xinjiang, and dated to the early Bronze Age, show excellent DNA preservation. Here we present an ancient DNA (aDNA) analysis of wheat (Triticum sp.) grains excavated from Xiaohe and provide the first definitive evidence for bread wheat in China during the Bronze Age. The nuclear ribosomal DNA internal transcribed spacer regions (ITS1 and ITS2) and the intergenic spacer region (IGS) were amplified. The IGS region within the D genome of wheat has a 71 bp insertion that is absent from corresponding regions in the A and B genomes. The results showed that the Xiaohe wheat showed most sequence similarity to hexaploid bread wheat (Triticum aestivum), including the characteristic insertion into the D genome. The presence of bread wheat at the Xiaohe cemetery is discussed in relation to it having spread into Xinjiang by the Bronze Age, providing new insight into the origins of bread wheat in East Asia.     

Ancient DNA typing shows that a Bronze Age mummy is a composite of different skeletons

Excavations at Cladh Hallan, a Bronze Age-Iron Age settlement on South Uist in the Outer Hebrides off the west coast of Scotland, revealed the skeletons of two adults, a sub-adult and a child buried beneath the foundations of three roundhouses. Osteological and isotopic evidence has shown that the male adult skeleton is a composite made up of parts of at least three different individuals. To test the hypothesis that the female skeleton was also a composite we examined ancient DNA from four of its components: the skull, mandible, right humerus and right femur. Seven polymerase chain reactions (PCRs) were attempted, these covering positions 15,996–16,420, 16–132 and 232–368 of the mitochondrial DNA hypervariable I and II regions. Three PCRs were successful for each sample and a total of 55 sequences were obtained from the cloned products. After exclusion of possible contaminating sequences, the remaining 34 were compared. It was concluded that the mandible, humerus and femur come from different individuals. Insufficient data were obtained to draw conclusions regarding the origin of the skull. The presence of two composite skeletons at Cladh Hallan indicates that the merging of identities may have been a deliberate act, perhaps designed to amalgamate different ancestries into a single lineage.     

Grave Circle B at Mycenae in the Context of Links Between the Eastern Mediterranean and Scandinavia in the Bronze Age

Artifacts from royal burial graves Gamma and Omicron of grave circle B at Mycenae attest to cultural ties between the Eastern Mediterranean elite and that of the Scandinavian Early Bronze Age (mid- and late 2nd millennium BC). The appearance of the running spiral motif and representations of ships with rams in Scandinavia coincide with the beginning of the Mycenaean civilization. These facts, along with the ?nds of Baltic amber only in the royal burials at Mycenae but not in Crete, suggest that a principal role in the introduction of these cultural elements in Scandinavia during the Scandinavian Bronze Age (periods I–III according to Montelius) was played by the Mycenaean elite.     

The limits of biomolecular palaeopathology: ancient DNA cannot be used to study venereal syphilis

To determine whether ancient DNA (aDNA) can be used to study the palaeopathology of venereal syphilis, we carried out a comprehensive analysis of the preservation of human and pathogen DNA in a set of 46 bones of various ages, most of which displayed osteological indications of the disease. Bones came from seven English cemetery sites that were in use during the 9th–19th centuries. Twelve of the 46 bones consistently yielded mitochondrial DNA (mtDNA) sequences after replicate polymerase chain reactions (PCRs), and a further 13 bones yielded mtDNA sequences with less reproducibility. The sequence data enabled tentative mitochondrial haplogroups to be assigned to nine of the bones, and the identities and frequencies of these haplogroups were compatible with the geographical origins of the bones. Twenty-one bones consistently gave negative results with all mtDNA PCRs, indicating that at least these bones were not contaminated with modern human DNA, and those bones that gave positive results only yielded one sequence each, again suggesting that widespread modern contamination had not occurred. Mycobacterium tuberculosis sequences were obtained from seven bones, including three of five bones with tuberculous lesions. The cloned and direct sequences obtained from both the mtDNA and M. tuberculosis PCR products showed features typical of degraded aDNA. All of these results suggest that at least some of the 46 bones that we studied were suitable for aDNA analysis. All 46 bones were tested with nine different treponemal PCRs, each optimised to give a detection limit of ≤5 genomes. Although various bones gave PCR products of the expected size with one or more of these PCRs, sequencing showed that none of these products were authentic treponemal amplicons. Our failure to detect treponemal DNA in bones that were suitable for aDNA analysis, using highly sensitive PCRs, suggests that treponemal DNA is not preserved in human bone and that it is therefore not possible to use aDNA analysis to study venereal syphilis. Any past or future paper claiming detection of treponemal aDNA should therefore be accompanied by a detailed justification of the results.     

Improved methodology for extraction and amplification of DNA from single grains of charred wheat

Comparisons were made between two commonly used methods for the extraction of ancient DNA from charred plant remains. Using artificially charred wheat seeds, we show that silica-binding is the most efficient method for extraction of DNA. We describe a improved silica-binding procedure, including pre-incubation with N-phenacylthiazolium bromide and increased washing of bound DNA, which yields amplifiable DNA from seeds heated at 200 °C for up to 8 h, conditions which promote the formation of Maillard products which often copurify with aDNA and inhibit subsequent PCRs. We believe that this method will be effective in ancient DNA extraction with most types of charred archaeobotanical material. Both cold- and hot-start PCR procedures gave good amplicon yields with extracts prepared in this way, but cold-start PCRs also resulted in synthesis of short artefact products. Addition of bovine serum albumin to PCRs, an inert carrier substance thought to enhance amplification efficiency by binding contaminants, had no advantageous effect and in fact reduced amplicon synthesis.     

The technology Of the yamnaya (pit grave) ceramic production And its relevance to the population history Of the volga-ural region in the early bronze age

Based on ceramic assemblages from the Yamnaya (Pit Grave) culture burials, the technology of pottery manufacture practiced by the Early Bronze Age people of the Middle Volga–Southern Ural region is described. The analysis follows the methodology developed by A.A. Bobrinsky. The results are used for reconstructing the origin and evolution of the Pit Grave culture. The role of ceramics in Early Bronze Age funerary ritualism is discussed.     

Post-mortem DNA damage hotspots in Bison (Bison bison) provide evidence for both damage and mutational hotspots in human mitochondrial DNA

Post-mortem damage-driven mutations are a phenomena associated with ancient DNA (aDNA) studies. A previous study has demonstrated that the distribution of such mutations in human mitochondrial DNA is not random, but is concentrated in ‘hotspots’ that correlate with sites of elevated mutation rate in vivo. However, as the previous study was undertaken on human samples, it is possible for a critic to argue that the results might be biased through the presence of modern contaminant DNA sequences among the ancient DNA extracts. In this study we confirm the phenomena of DNA damage hotspots using a data set that is unlikely to be affected by contamination – cloned mitochondrial control region sequences extracted from 81 ancient bison (Bison bison). Furthermore, using published data from modern bovid specimens, we confirm that the damage hotspots correlate with sites of in vivo hypermutation. In conclusion, the aDNA sequences from archaeological specimens provide evidence that structural elements of mitochondrial DNA confer a degree of in vivo and post-mortem protection from sequence modification. This in turn provides useful insights into the debate as to whether mutational hotspots or mitochondrial recombination might best explain homoplasies observed on phylogenetic trees of human mitochondrial sequences.     

COMPLEX RELATIONSHIPS BETWEEN MITOCHONDRIAL AND NUCLEAR DNA PRESERVATION IN HISTORICAL DNA EXTRACTS*

‘Historical’ DNA obtained from specimens preserved in natural history collections has proven useful for addressing a wide variety of questions, such as the spread of domesticated species or changes in genetic diversity. With the development of high‐throughput sequencing techniques, there is an increasing focus on acquiring genetic information encoded by single‐copy nuclear DNA from historical DNA extracts. The development of efficient techniques to determine the level of nuclear DNA preservation in candidate specimens is necessary to maximize the data obtained from these analyses. Although current evidence suggests that a sample's mitochondrial DNA preservation predicts its nuclear DNA preservation, we show that the relationships between mitochondrial and nuclear DNA preservation are complex.     

I Will Follow You into the Dark: Death and Emotion in a Mycenaean Royal Funeral

Despite many years of intensive research into burial and funeral practices in Late Bronze Age (LBA) Greece, emotion remains largely absent from the discussion. Yet death and the emotions it provoked would have been familiar aspects of daily life in Mycenaean Greece. The dead had to be dealt with and moved on through various rites until they became a safe part of the landscape and memory of individuals and communities. In addition to anthropological and psychological research on death and grieving, we have several categories of evidence that can allow us a glimpse of the emotional world of death in LBA Greece: the Homeric poems the Iliad and Odyssey, iconographical representations of death, funeral practices and mourning, and the archaeological material itself – the tombs and offerings. In this paper, I introduce the various categories of evidence and draw on them to support an imaginative reconstruction of an event that happened, but which is not recorded in any historical sources – the death and burial of a great king of Mycenae. I argue that using emotion as a lens through which to view the evidence can sometimes allow deeper interpretation and enable fuller historical reconstructions of lived lives and experiences.     

THE ORIGIN OF THE PICTORIAL KRATER FROM THE ‘MYCENAEAN’ TOMB AT TEL DAN

A Late Bronze Age Mycenaean pictorial krater, decorated with a chariot procession, from tomb 387 at Tel Dan, Israel, was examined by instrumental neutron activation analysis. The origin of the krater is placed in the Argolid, very likely in the region of Mycenae or Berbati.     

DNA in cremated bones from an early bronze age cemetery cairn

The discovery of polymeric DNA in charred wheat grains and other burnt plant remains prompted us to examine a set of cremated bones for the presence of ancient DNA. Extracts of cremated bones from an early Bronze Age cemetery cairn were analysed by hybridization probing and the polymerase chain reaction. Hybridization with two probes targeting human DNA resulted in strong positive signals. Polymerase chain reactions directed at a multicopy sequence in human nuclear DNA were successful, extracts of five burials giving amplification products of the expected size. Exploitation of ancient DNA in cremated bones could be of great value in archaeology as it may enable information pertaining to sex and kinship to be obtained from fragmented material.     

A provenance study of Mycenaean pottery from Northern Israel

The occurrence of imported Mycenaean pottery in the Late Bronze Age southern Levant is one of the most conspicuous aspects of Eastern Mediterranean trade connections during this period. A group of 183 Mycenaean pottery vessels from 14 sites in northern Israel, from both coastal and inland settlement contexts were analyzed by Neutron Activation Analysis. The results indicate that the vast majority of these vessels have a similar profile and can be provenanced to the north-eastern Peloponnese or more specifically, to the Mycenae/Berbati workshop in the Argolid. Possible interpretations of these results are presented and discussed against the historical and cultural background of the Late Bronze Age Eastern Mediterranean.     

Sheep wool in Bronze Age and Iron Age Europe

This study presents the results of a series of wool measurements from Bronze Age and Iron Age skins and textiles from Hallstatt, and Bronze Age textiles from Scandinavia and the Balkans. A new method of classification that was set up and applied on mostly mineralised Iron Age material has now been applied to a large body of non-mineralised material from the Bronze and Iron Ages. Three types of microscopes were used and their advantages and disadvantages assessed. The results of the investigation cast new light on sheep breeding and fibre processing in prehistoric Europe, and suggest that different sheep breeds existed in Bronze Age Europe.     

Neolithic and Bronze Age Agriculture in Southern Scandinavia – Recent Archaeobotanical Evidence from Denmark

Abstract

This paper presents archaeobotanical studies from the Danish regions of Thy, northern Schleswig and Djursland. The data are discussed in the light of developments in the landscape and in house architecture; comparisons are made with the contemporary situation in southern Sweden. Pollen analysis reveals that Thy was more or less treeless by the end of the Neolithic, whereas Djursland maintained its forests for a further 1500 years; the situation in northern Schleswig lies somewhere in between. Developments in house architecture are very similar in the three areas. The shift from two-aisled to three-aisled houses occurred in period I/II of the Bronze Age and phosphate analyses suggest that the earliest Danish byre dates from the beginning of period II.

Crop plant assemblages are dominated by naked barley and emmer and remain remarkably stable from the Single Grave Culture to the Late Bronze Age in Thy, from the Middle Neolithic to the middle of the Bronze Age in northern Schleswig, and from the Late Neolithic to the Late Bronze Age on Djursland. Other crops come and go – einkorn, bread wheat, spelt, millet, flax, oats and gold of pleasure. Hulled barley is largely conspicuous by its absence. Well-developed arable weed floras appear first in the Early Bronze Age – arable weeds are very scarce at Neolithic sites. There is evidence of improvement of arable soils using fen peat and household refuse and manure. The situation appears somewhat more complex in Denmark than that described for Sweden. The most striking difference is seen in the behaviour of hulled barley, which becomes massively dominant in Sweden in the course of the Bronze Age, whereas its role in Denmark is much more modest.     

An Analysis Of Mitochondrial Dna From The Pakhomovskaya Population Of The Late Bronze Age,Western Siberia

This article presents the results of an analysis of mitochondrial DNA extracted from bone samples from Stary Sad – a burial ground representing the eastern variant of the Late Bronze Age Pakhomovskaya culture in the Baraba forest-steppe, Western Siberia. Comparison with mitochondrial DNA data from earlier populations of the region and also with archaeological facts, points to the origins of the Pakhomovskaya people. Certain components of their gene pool were evidently derived from the local pre-Andronovo populations, others from the actual Andronovo (Fedorovka) population and also from later immigrants. In this article an integrative reconstruction based on biological and cultural facts is proposed.     

On the Trail of the Oldest Domestic Cat in Poland. An Insight from Morphometry,Ancient DNA and Radiocarbon Dating

It is believed on the basis of archaeozoological research that the domestic cat appeared in Central Europe during the Roman Period. In Poland, the domestic cat is a common species in medieval deposits. Only a few finds of cat remains of pre‐medieval age have been reported from Poland to date, including several specimens from deposits older than the Roman Period, dated to the pre‐Roman Period and even the Bronze Age. To clarify the earliest history of the domestic cat in Poland, the paper presents a review of the available published cat remains and adds some data about newly discovered remains. Combined methods of morphometry and ancient DNA were applied to enable distinction of wildcats and domestic cats. The domestic cat remains were radiocarbon dated. In six cases of domesticated cat reported in the literature, five were positively taxonomically verified, both by morphology and by genetic analysis, and one was recognised as a European wildcat. According to radiocarbon chronology, the oldest studied find is dated to the fourth–third century bc and represents a wildcat. Only two individuals of domestic cat – skeletons from Łojewo and Sławsko Wielkie, both from Kuyavia region (central‐northern Poland) – represent the Roman Period (first–third century ad ), and they are the oldest confirmed domestic cats in Poland. Copyright © 2015 John Wiley & Sons, Ltd.     

Ancient DNA analysis on Clonorchis sinensis eggs remained in samples from medieval Korean mummy

In the present study, Clonorchis sinensis (C. sinensis) ancient DNA (aDNA) was successfully extracted from human remains discovered in a tomb dating to the medieval Joseon dynasty of Korea. The presence of C. sinensis eggs was confirmed by microscopic observation, after which a PCR-based aDNA analysis was performed using primer sets designed for the amplification of nuclear ribosomal internal transcribed spacer 1 (ITS1), 2 (ITS2) and mitochondrial cytochrome c oxidase 1 (CO1) genes. The sequences obtained were 100% homologous to some contemporary C. sinensis gene sequences reported from Korea and other East Asian countries. We believe that the results of our analysis expand the temporal and geographical scopes of research on the history of C. sinensis infection in different human populations.     

The survival of PCR-amplifiable DNA in cow leather

We have investigated the survival of PCR-amplifiable mitochondrial and nuclear DNA in a small number of modern and medieval bovine leather samples. The results of this preliminary investigation demonstrate that, while no nuclear DNA can be PCR-amplified from any of the specimens, mitochondrial DNA can be amplified from all samples. To investigate this contrasting pattern of DNA survival further, we have vegetable-tanned cow skin in our own laboratory, and directly assayed the survival of PCR-amplifiable mitochondrial and nuclear DNA at each step of the process. The results indicate that nuclear DNA is reduced to sub-amplifiable levels as a result of the tanning baths, whereas amplifiable mitochondrial DNA survives the complete process. Our results suggest that old and archaeological bovine leather may represent a useful source of genetic information, although this information will most likely be limited to that which can be gained from mitochondrial DNA.     

Evidence of Social Stratification in Bronze Age Denmark

Abstract

The Late Bronze Age hoards (12th–6th centuries B.C.) from Denmark are examined as evidence of the existence of social ranking in that prehistoric society. The hoards contain bronze weapons and ornaments which seem to function as sumptuary goods and appear to be ranked according to regular rules. The hoards also represent economic wealth and include objects of ritual importance. This intersection, in single finds, of material reflections of the political, religious, and economic systems in the society, along with the inferred existence of social ranking, suggests the presence of a prehistoric chiefdom in Denmark in the Late Bronze Age.