Mitochondrial DNA Analysis of Ancient Domestic Goats in the Southern Caucasus: A Preliminary Result from Neolithic Settlements at Göytepe and Hacı Elamxanlı Tepe

This study presents preliminary results of mitochondrial DNA analyses of modern and ancient domestic goats in the southern Caucasus in order to examine their phylogenetic relationship with modern and ancient goats. For this purpose, seven ancient samples were obtained from two early agricultural villages in west Azerbaijan (Göytepe and Hacı Elamxanlı tepe, dated to ca. 6000–5500 cal bc , the Pottery Neolithic period), in addition to five modern goat samples in the same region. In the study, mitochondrial DNA segments of the control region (216 bp for the Neolithic samples and 481 bp for the modern samples) were amplified, and phylogenetic analyses were performed using previously published reference DNA sequences. As a result, all the haplotypes found in this study were grouped in the haplogroup A of goats. The finding of the haplogroup A among domestic goats in the southern Caucasus in the early sixth millennium bc can be interpreted as part of the geographic expansion of this lineage from the areas of initial domestication to surrounding areas that include also South and Southeast Europe. In the southern Caucasus, the haplogroup A probably continued to be a major lineage among domestic goats since their emergence in this area to the present. In contrast, this lineage has not been detected among local wild goats including Capra aegagrus , indicating the external origin of domestic goats. This possibility is consistent with archaeological records that indicate sudden appearance of agricultural lifeways in the southern Caucasus and cultural connections with northern Mesopotamia. Copyright © 2016 John Wiley & Sons, Ltd.     

Ancient DNA,a Neolithic legging from the Swiss Alps and the early history of goat

Ancient DNA from a Neolithic legging (1st half of the 3rd millennium BC) found at Lenk, Schnidejoch (2750 m a.sl.) in the Swiss Alps has demonstrated, that modern distribution of genetic variation does not reflect past spatio-temporal signatures. The legging was made from the skin of a domestic goat (Capra hircus), belonging to the caprine haplogroup B1, which is marginal in Europe today, but represents a third highly diverse goat haplogroup entering Europe already in the Neolithic. Population expansion of lineage B therefore happened more than 4500 years ago, but their members were at some point almost completely replaced by goats of today's common A and C haplogroups.     

Ancient DNA provides new insights into the origin of the Chinese domestic horse

Domestic horses played a pivotal role in ancient China, but their exact origin remains controversial. To investigate the origin of Chinese domestic horses, we analyzed mitochondrial DNA (mtDNA) from 35 horse remains, aged between 4000 and 2000 years, excavated from nine archaeological sites in northern China. The Chinese ancient horses exhibited high matrilineal diversity, falling into all the seven haplogroups (A–G) observed in modern horses. These results suggest that several maternal lines were introduced into the gene pool of Chinese horses in the past. Haplogroups A and F were more prevalent in ancient horses than the other haplogroups. Interestingly, only haplogroups A and F were present in the samples older than 4000 years, while the more recent horses (between 2000 and 3000 years BP) fell into all seven haplogroups. Comparison with DNA data of present-day horses suggests that haplogroup F is like to be an ancient haplogroup of East Asian origin. These analyses also suggest that the origin of Chinese domestic horses is complex, and external mtDNA input occurred after initial domestication. Our results indicate that the Chinese ancient horses are more related to the modern Mongolian horses. Lastly, our results cannot support the previous hypothesis that early Chinese domestic horses were derived from the Przewalski horse.     

Mitochondrial DNA diversity in a Transbaikalian Xiongnu population

Xiongnu was a confederation of nomadic pastoral tribes (~200 bc–100 ad) that founded the first nomadic empire in Central Asia. According to archeological and historical data, the tribes played a key role in ethnic and cultural processes in Central Asia and adjacent regions of Eurasia. Genetic studies of the Xiongnu published to date have focused on remains from burial grounds in present-day Mongolia, in the southern part of the ancient Xiongnu area. However, paleoanthropological materials from numerous Xiongnu cemeteries and settlements in Transbaikalia (the southern region of Eastern Siberia, Russia) in the northern part of the Xiongnu Empire have not been examined genetically. Here, we analyzed mitochondrial DNA variation in a Transbaikalian Xiongnu population based on ancient DNA obtained from skeletal remains (n = 18) at four burial grounds to complement available Xiongnu genetic diversity data. We detected 16 mitochondrial DNA haplotypes belonging to seven East Eurasian haplogroups (A, B5, C, D4, G2a, N9a, and Y) in the Transbaikalian Xiongnu series. We observed substantial similarity between Transbaikalian and Mongolian Xiongnu series with respect to main haplogroup composition and frequencies. We observed several mitochondrial DNA clusters (N9a, Y, B5, and A16) and 11 of 16 haplotypes that were previously undetected in the Xiongnu gene pool. We also observed high similarity between the Xiongnu and contemporary indigenous populations of eastern Central Asia, particularly Mongolian-speaking groups. These findings extend our knowledge of Xiongnu genetic diversity.     

Mitochondrial haplogroup M discovered in prehistoric North Americans

We analyzed two mid-Holocene (∼5000 years before present) individuals from North America that belong to mitochondrial DNA (mtDNA) haplogroup M, a common type found in East Asia, but one that has never before been reported in ancient or living indigenous populations in the Americas. This study provides evidence that the founding migrants of the Americas exhibited greater genetic diversity than previously recognized, prompting us to reconsider the widely accepted five-founder model that posits that the Americas were colonized by only five founding mtDNA lineages. Additional genetic studies of prehistoric remains in the Americas are likely to reveal important insights into the early population history of Native Americans. However, the usefulness of this information will be tempered by the ability of researchers to distinguish novel founding lineages from contamination and, as such, we recommend strategies to successfully accomplish this goal.     

Wild or domesticated: DNA analysis of ancient water buffalo remains from north China

Recent zooarchaeological studies on water buffalo (Bubalus sp.) remains from China and south Asia question the traditional view that water buffalo were first domesticated in Neolithic China over 7000 years ago. The results from several recent population genetic studies of modern domesticated buffalo (Bubalus bubalis) are not consistent with each other, placing the original center of buffalo's domestication in south Asia, southeast Asia, or China. This paper reports a study using an ancient DNA approach to analyze water buffalo remains from Neolithic sites in north China to investigate their affinities with modern domesticated water buffalo, and to shed light on the origin of modern domesticated water buffalo in China.A 169 bp fragment of D-loop mitochondrial DNA was successfully amplified and verified for 13 of 24 bone samples obtained from seven archaeological sites along the Wei River valley in Shaanxi Province, China. The bone samples which yielded positive DNA can be dated to 8000–3600 cal. BP. The phylogenetic analysis of the obtained DNA sequences along with modern water buffalo sequences indicated that the ancient water buffalos were not the direct ancestor of modern domesticated water buffalo. However, the phylogenetic analysis, along with BLAST searches of these ancient DNA sequences, did demonstrate their relatedness to water buffalo more so than to any other bovid species, confirming the existence of indigenous wild (but now extinct) water buffalo species (B. mephistopheles) in ancient China.The DNA analysis of these ancient remains failed to establish direct links between modern domesticated water buffalo (B. bubalis) and indigenous water buffalo (B. mephistopheles) from ancient China. If further DNA studies of more ancient remains from other regions of China confirm the observation of solely indigenous water buffalo species in ancient China, it would suggest modern water buffalo might not have been first domesticated in China.     

Ancestral Puebloan mtDNA in context of the greater southwest

Ancient DNA (aDNA) was extracted from the human remains of seventy-three individuals from the Tommy and Mine Canyon sites (dated to PI-II and PIII, respectively), located on the B-Square Ranch in the Middle San Juan region of New Mexico. The mitochondrial DNA (mtDNA) haplogroups of 48 (65.7%) of these samples were identified, and their frequency distributions were compared with those of other prehistoric and modern populations from the Greater Southwest and Mexico. The haplogroup frequency distributions for the two sites were statistically significantly different from each other, with the Mine Canyon site exhibiting an unusually high frequency of haplogroup A for a Southwestern population, indicating the possible influence of migration or other evolutionary forces. However, both sites exhibited a relatively high frequency of haplogroup B, typical of Southwestern populations, suggesting continuity in the Southwest, as has been hypothesized by others (8, 9, 29, 45 and 57). The first hypervariable region of twenty-three individuals (31.5%) was also sequenced to confirm haplogroup assignments and compared with other sequences from the region. This comparison further strengthens the argument for population continuity in the Southwest without a detectable influence from Mesoamerica.     

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.     

DNA analysis of archaeological sheep remains from China

Recent research has thrown considerable light on the history of the domestic sheep, but has not extended to ancient sheep specimens. In the present study, ancient DNA analysis was carried out on eight archaeological sheep remains recovered from Erlitou archaeological site in Henan Province (ca. 2100–1800 B.C.) to explore the genetic structure of ancient sheep and the phylogenetic relationship between ancient and modern sheep. We analyzed the control region sequences and coding regions of mitochondrial DNA from the remains by direct sequencing and restriction fragment length polymorphism analysis, respectively. Our results reveal that all ancient sheep belong to lineage A defined by modern sheep sequences. Phylogenetic analysis shows that neither argali (Ovis ammon) nor urial (Ovis vignei) mtDNA is closely related to Erlitou ancient sheep. In addition, our results suggest that ancient DNA analysis can serve as a powerful tool in tracing prehistoric population movement.     

First ancient bovine DNA evidence from India: difficult but not impossible

Ancient DNA isolation from the tropical countries has been shown to be very difficult in the past. Here for the first time we have been successful in isolating ancient DNA from Indian cattle samples. We were able to obtain DNA and sequence the partial mitochondrial D-loop in 3 of the 15 bovine fossil samples ranging in age from 2000 BC to 1000 AD, and were able to further identify the most recent sample as being of Bos indicus origin. Our results on ancient DNA extraction from India will encourage other researchers in this field to carry out further studies of ancient DNA from Indian bovine samples. Our results represent the first successful extraction and amplification of bovine ancient DNA from India, and thus may pave the road for a better understanding of demographic and historical processes of cattle domestication that has taken place in this region.     

The relationship of the Mimbres to other southwestern and Mexican populations

Previous aDNA research in the Southwest provides a framework to understand the Mimbres population which maintains a unique identity in the archeology of the desert Southwest of North America. Both the mitochondrial DNA (mtDNA) haplogroups and haplotypes of a sample of 46 Mimbres individuals were identified from skeletal remains and compared to those of other ancient and modern populations in the American Southwest and Mexico. The haplogroup frequency distribution of the Mimbres resembles that of other Southwestern populations, suggesting a close relationship between the Mimbres and the neighboring populations and population continuity for a long period of time. However, their relatively low frequency of haplogroup A (11% n = 5) is consistent with a clinal distribution of haplogroup A from Mesoamerica to the Southwest. Moreover, unlike other Southwestern populations, the Mimbres share haplotypes with populations to the south, such as the Cora, Huichol, and Nahua, suggesting gene-flow from Mesoamerica. Here we discuss this data in light of the origin of the Mimbres population and their relationship to other Southwestern and Mexican populations.     

Holey Goats: Multiple Cases of Supratrochlear Foramina in the Humerus of Caprines from the New Kingdom Pharaonic Town of Amara West,Northern Sudan

Supratrochlear foramina (STF) were recorded in fifteen per cent of goat and sheep/goat humeri from the New Kingdom pharaonic town of Amara West, in modern northern Sudan. To the authors’ knowledge, this trait has never before been reported in the published literature for goats or sheep, whether from archaeological or modern contexts. The aim of this work is twofold: to contribute to the growing corpus of studies addressing the incidence and aetiology of STF, and to raise awareness for their possible presence in caprines, thus encouraging their identification and recording in archaeological assemblages.     

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.     

Ancient DNA insights from the Middle Neolithic in Germany

Genetic studies of Neolithic groups in central Europe have provided insights into the demographic processes that have occurred during the initial transition to agriculture as well as in later Neolithic contexts. While distinct genetic patterns between indigenous hunter-gatherers and Neolithic farmers in Europe have been observed, it is still under discussion how the genetic diversity changed during the 5,000-year span of the Neolithic period. In order to investigate genetic patterns after the earliest farming communities, we carried out an ancient mitochondrial DNA (mtDNA) analysis of 34 individuals from Wittmar, Germany representing three different Neolithic farming groups (ca. 5,200–4,300 cal bc) including Rössen societies. Ancient DNA analysis was successful for six individuals associated with the Middle Neolithic Rössen and observed haplotypes were assigned to mtDNA haplogroups H5, HV0, U5, and K. Our results offer perspectives on the genetic composition of individuals associated with the Rössen culture at Wittmar and permit insights into genetic landscapes in central Europe at a time when regional groups first emerged during the Middle Neolithic.     

Characterising the potential of sheep wool for ancient DNA analyses

The use of wool derived from sheep (Ovis aries) hair shafts is widespread in ancient and historic textiles. Given that hair can represent a valuable source of ancient DNA, wool may represent a valuable genetic archive for studies on the domestication of the sheep. However, both the quality and content of DNA in hair shafts are known to vary, and it is possible that common treatments of wool such as dyeing may negatively impact the DNA. Using quantitative real-time polymerase chain reaction (PCR), we demonstrate that in general, short fragments of both mitochondrial and single-copy nuclear DNA can be PCR-amplified from wool derived from a variety of breeds, regardless of the body location or natural pigmentation. Furthermore, although DNA can be PCR-amplified from wool dyed with one of four common plant dyes (tansy, woad, madder, weld), the use of mordants such as alum or iron leads to considerable DNA degradation. Lastly, we demonstrate that mtDNA at least can be PCR-amplified, cloned and sequenced from a range of archaeological and historic Danish, Flemmish and Greenlandic wool textile samples. In summary, our data suggest that wool offers a promising source for future ancient mitochondrial DNA studies.     

Genetic analysis of archaeological wood remains: first results and prospects

A total of 51 ancient oak wood samples originating from various European archaeological sites, dating from the Neolithic period to the 18th century, were assayed for the presence of reproducible chloroplast (cp) DNA sequences. Five polymorphic chloroplast fragments were targeted. Only five of the samples could be fully genetically characterised, revealing four different oak cpDNA haplotypes. In all cases, the haplotypes detected on ancient woods and the haplotypes characterised from fresh samples from the same localities matched. Overall, this congruence is consistent with a genetic continuity between ancient and modern European oaks, confirming the hypothesis that the mapped genetic patterns largely reflect the original structure that established during the post-glacial. This stability of the genetic structure implies that, in the future, the technique could be used to infer or confirm the transport of wood by man, providing interesting perspectives for the genetic analysis of ancient woods.     

Ancient DNA evidence for genetic continuity in arctic dogs

The domestic dog fulfills many functions for their human companions, such as hauling, guarding, and protection. Consequently, humans have taken dogs to nearly every corner of the globe. Recent translocations of Western dogs stemming from the Victorian era dog fancy have erased some of the ancient genetic signatures of these earlier migrations. Here we used DNA of ancient and modern dogs from the North American Arctic of Alaska and Greenland to assess their genetic continuity in time and space. We successfully sequenced 23 archaeological (ca. AD 1250–1910) and recent (ca. AD 1930–1990) surface-collected Canis bone and tooth samples and compared them to 51 modern Inuit Sled Dogs and to published sequences of modern Alaskan Malamutes (and additional Inuit Sled Dogs) to test for evidence of lineage replacement or genetic continuity through time. Ancient samples from Alaska and Greenland and modern sequences from Greenland all contained a high frequency of haplotype A31, which was previously described only in modern North American Arctic dogs. Thus, A31 was a common thread tying the entire North American Arctic together prior to European colonization and, in the Eastern Arctic, indicates genetic continuity between past and present dogs as well. However, A31 is rare in modern Alaskan dogs, consistent with post-colonization replacement by Eurasian matrilines.     

Minimally destructive DNA extraction from archaeological artefacts made from whale baleen

Here we demonstrate the successful extraction and amplification of target species DNA from artefacts made of whale baleen collected from excavations of past palaeo-Eskimo and Inuit cultures in Greenland. DNA was successfully extracted and amplified from a single baleen bristle of 1.5 cm length dated based on archaeological context to the period of the Saqqaq culture, more than 4000 years ago and following decades of storage at room temperature at the National Museum. The results reveal ancient baleen in archaeological material as a potential source of DNA that can be used for population genetic studies. We conclude that genetic investigation of historical baleen collections can contribute to our knowledge of the prehistoric population genetics of baleen whales, for example by quantifying the impact of modern whaling on the genetic diversity of bowhead whales.     

DNA FROM HUMAN ANCIENT BACTERIA: A NOVEL SOURCE OF GENETIC EVIDENCE FROM ARCHAEOLOGICAL DENTAL CALCULUS

We report a molecular methodology to obtain and analyse ancient bacterial DNA from archaeological dental calculus. Recent and archaeological DNA samples, as old as 4000 bp , were successfully extracted and amplified with species‐specific PCR primers. We propose this approach in order to: detect the presence of specific bacterial species infecting past human populations; compare the composition of ancient oral microbiomes among human populations; and analyse the genetic variability and covariation of bacteria and human host populations. Genomic analysis of bacteria from dental calculus is a promising source of evidence for palaeopathological and micro‐evolutionary studies, focused either on micro‐organisms or their human hosts.