Archaeological DNA data of the XII century from ancient Klyazma settlements. Part 2

Semenov Alexander Sergueevich PhD in Physics and Mathematics CEO, project "DNA-history of Russia" 119334, Russia, Moscow, ul. Leninsky Prospekt, 43, sq. 161 semyonov1980@mail.ru Other publications by this author     Kabaev Danil Andreevich Leading Specialist, LLC “Vladimir Region Center for Archaelogy of the Vladimir State University” 600009, Russia, Vladimir region, Vladimir, Mira str., 9 d.kabaev@mail.ru Other publications by this author     Chernyaeva Larisa Leonidovna CEO, LLC "Vladimir Region Center for Archaelogy of the Vladimir State University" 600009, Russia, Vladimir region, Vladimir, Mira str., 9 galchuk@list.ru Other publications by this author     Chernov Serguey Zaremovich Doctor of History Leading Researcher, Moscow Rus' Archaeology Department, Institute of Archaeology of the Russian Academy of Sciences 117292, Russia, Moscow, Moscow, ul. D. Ulyanova, 19 chernovsz@mail.ru Other publications by this author

Goncharova Natalia Nikolaevna PhD in Biology Assistant Professor, Faculty of Biology, Moscow State University 119234, Russia, Moscow, Moscow, Leninskye Gory str., 1/12 1455008@gmail.com Other publications by this author

paleoDNA testing is becoming one of the most complex areas of modern biotechnology. Modern technologies make it possible to qualitatively extract DNA and test it by various methods, including the determination of the Y-chromosome haplogroup and mitochondrial DNA ^[1]. This method of applied historical research has been implemented for 20 years, but data on Slavic and other populations of the era of the birth of the Russian state are still few. The review is given in [2-6,10-14]. We know the DNA of no more than 20 ancient Russian burial grounds, so at the current stage there is a primary accumulation of information. Now, for the territory of our country, the stage of initial "data mining" is still ongoing and the first generalization can be made after 40-50 studies. Thus, the formation of a list of data on the ancient DNA of the largest burial grounds within the borders of Ancient Russia with a sample of ancient DNA is relevant.

The main purpose of this study was to determine the haplogroup in two medieval graves of the XII century of the middle course of the Klyazma River and to search for their possible analogues in other medieval DNA samples. Two graves were chosen as the object, examined in 2016-2017 at the medieval cemetery of the XII-XIV centuries in Vladimir, the necropolis is located on the territory of the Patriarchal Garden complex. This part describes burial No. 25 of the XII century. Burial No. 26 was described in the first part of this work ^[14]. The material for research was obtained as a result of archaeological excavations in the area of reconstruction of the Patriarch's Garden complex. The burial ground and the adjacent settlement are located in the southwestern part of the historical center of Vladimir, on the slope of the floodplain terrace on the left bank of the Klyazma River. The excavations were carried out by employees of LLC "Vladimir Regional Center of Archeology at VlSU" S.V. Ocheretina and D.A. Kabaev. In total, about 180 burials were examined on an area of more than 1000 m2.

The new (or Earthen) city begins to form in the XII century, the construction of its fortifications dates back to 1158-1164 under Andrei Bogolyubsky. Active development of this part of the city begins in the XII-XIII centuries. The Church of the Savior is being built here, then the Spaso-Zlatovratsky Monastery appears, which existed until 1784. In the XIV-XV centuries. in the western part of the studied area there were craft workshops for the production of iron locks and keys to them. Then here, on the southern slopes of the ravine, the Patriarchal Garden appears, which was assigned to the Nativity Monastery and had different names – Episcopal, Synodal, Patriarchal, depending on the subordination of the monastery itself.

The burial ground, discovered during excavations, dates from clothing and ceramic material, its functioning falls on the period of the XII-XIV centuries. Most of the burials are uninventional, but in the earliest burials there are elements of clothing that allow dating these burials with a narrower time interval.

The burial complex of the XII century is particularly distinguished, and in it there is a female burial No. 25. Among the details of the burial clothes from burial No. 25, elements of gold–woven clothing were found - a silk ribbon with an ornament, a fragment of a gold-woven collar with three bronze buttons and a birch bark lining, possibly a bib with gold thread embroidery on wool and silk fabric. Similar items were found during excavations in the cities of Dnieper Russia and in the burial mounds of Northeastern Russia, and date back to the XI-XIII centuries ^[15].

Gold-woven ribbons were in use among the population of Ancient Russia during the XI-XIII centuries, they were sheathed at the collar of the dress and sleeve cuffs. These products are most likely of Byzantine origin, most of the items of this kind were Byzantine. Despite the idea that gold-woven clothing was a rarity, M.V. Fechner writes that more than 80% of finds of this kind come from ordinary burial mounds of rural and urban population of Ancient Russia ^[16].

Copper mushroom-shaped buttons are found in most ancient Russian cities. In the monuments of Vladimir-Suzdal Rus, they belong to the pre-Mongol time and date from the XI-beginning of the XIII century ^[17].

Anthropological material from the burial is rather scarce. Small fragments of the skull and long leg bones have been preserved. It is impossible to establish the lifetime length of the body from the available fragments of long bones, the bone material allows us to say only that the bones belong to a woman, the age of death is 25-30 years.  The face is low, the nose is relatively wide, but at the same time the nose protrudes quite strongly. The fragmented nature of the remains allows us to give only these small descriptions. All the more important is the information that the study of DNA data on burial can give in this case.

Burial No. 26 was located not far away. Burial No. 26 was oriented according to the Orthodox tradition with its head to the west. The hands are folded on the stomach and pelvis. It dates from the XII-XIII centuries and is described in the article ^[14].

The work on determining the mito-haplogroup of the ancient burial was carried out at DNA-Heritage LLC in September 2019 - July 2020 under contracts no.DNA-La/11-19 dated 08.11.2019 and no.DNA-La/08-20 dated 14.08.2020. The interpretation of the result was carried out by the authors.

Materials and methods

DNA from the teeth was extracted by the SiO ₂ column method. The concentration of the isolated DNA was measured on a Qubit 2 fluorimeter (Thermo Fisher Scientific, USA). 

The concentration of the sample taken for mito-DNA analysis from burial No. 25 was 1.12 ng/ml of powder weighing 1 g. As a result of NGS sequencing (MiSeq platform, Illumina (USA), according to Illumina protocols)  The following data were obtained from the HVR1 and HVR2 regions of mitochondrial DNA: HVRI - 16558A, 16566A and HVR2 - 146C, 263G. NGS sequencing was performed using PowerSeq CRM (Promega) reagents.

Data and interpretations

As a result of the work, the following data were obtained. Based on the coincidence of HVR1 with the Cambridge reference sequence at the site up to 16519C and the presence of two mutations in HVR2, it was concluded that the mitochondrial haplotype of a woman was assigned to haplogroup H1-146C with a high probability (a combination of CRS and the presence of 263G in HVR2). According to the FTDNA table of defining mutations (https://www.familytreedna.com/mtdna-haplogroup-mutations.aspx ), mutation 146C indicates the H1m1 branch.

 Discussion and analysis

GenBank databases (http://www.ianlogan.co.uk/sequences_by_group/h1m_genbank_sequences.htm ) They show that the H1 branch belongs to Central and Eastern Europe, and H1m belongs, rather, to the Central European region and the Western Baltic and the shores of the North Sea (carriers are known in modern Poland, Germany, medieval England). The West Baltic and West Slavic regions also belong to the subclades I1-Z58, found in burial No. 26 (analogous to the West Slavic burial of Krakauer Berg ^[14]).  This assessment correlates with the conclusions made for individual No. 26 about the probable roots from the territory of the southwestern Baltic.

The testing data of contemporaries conducted by the DNA-History of Russia project showed the rarity of the mitotype of burial No. 25 in the Center of Russia.  However, for several tested families from Central Russia, it was detected, for example, in carriers with roots from the village of Chermny in the Kadomsky district of the Ryazan region (Tamara Evdokimovna Chugunova, born in 1940, and, consequently, her mother Anisya Ilyinichna Larkina, born in 1897). This may indicate the preservation of this line among the modern population of the Volga-Oka interfluve.

 Conclusion

The article presents the results of the research conducted in 2019-2020. The first evidence of the possible presence of H1m1 and I1-Z58 among the Klyazma population of Northeastern Russia in the XII century not only confirms the opinion that the presence of mito-haplogroup H1 on all medieval Russian lands (inhabited by descendants of Eastern Slavs), but also indicates that a certain genetic unity with the western parts of the Slavic area could exist at that time (since the most serious concentrations of I1-Z58 and the presence of H1m1 are observed in the western part of the Slavic area and in even more western territories). This hypothesis was discussed in previous works of the authors ^[4,5,14] in the context of articles ^[7-9].

Thanks

This research was carried out at the expense of LLC "Vladimir Regional Center of Archeology at VLSU", the project "DNA-history of Russia", patrons V.P. Savransky, K. S. Neverova, V.A. Krupnov, V.A.Prokhorov.