Results of determination of the Y-DNA haplogroup for an individual from the Nikultsinsky burial ground of the Gavrilov-Yamsky district of the Yaroslavl region

Semenov Alexander Sergueevich PhD in Physics and Mathematics CEO, Deep Dive Group, project "DNA-history of Russia" 119334, Russia, Moscow, ul. Leninsky Prospekt, 43, sq. 161 semyonov1980@mail.ru Other publications by this author

Kazenkov Oleg Yur'evich Advisor to the Rector's Office, P.G. Demidov Yaroslavl State University 150003, Russia, Yaroslavskaya oblast', g. Yaroslavl', ul. Sovetskaya, 14 o.kazenkov@gmail.com

Introduction

In the present work on the study of the DNA of the burial found on the territory of  The Nikultsinsky burial ground (the famous burial ground of the Fatyanovo culture described in ^[1,2,3]) within the Gavrilov-Yamsky district of the Yaroslavl region on the right-bank plateau of theVondel, an attempt has been made to verify by Y-DNA analysis the hypothesis that this burial belongs to the Fatyanovo culture of the Bronze Age III-II thousand BC. At the moment, the monument belongs to the category of lost – destroyed by a quarry. Information about the burial and the possibility of conducting research was provided by the Associate Professor of the Faculty of History of the Yaroslavl State University named after P.G. Demidov Spiridonova E.V.

 Archaeological data

          The first finds at the Nikultsinsky monument were made in 1930 – three human bones and a stone drilled axe. In the early 1960s, two more burials and a drilled axe were found. Systematic excavations at the burial ground were conducted in 1964-65 by D.A. Krainov (Upper Volga expedition). Grave grave area of 3000 ^m2 was uncovered and 18 graves were found at a depth of 0.5-1.7 m. Many graves were partially destroyed. 5 male, 5 female and 6 children's graves were investigated. In two graves there were paired burials: a female with a child and a double child (a boy with a girl).  The burial inventory includes stone navicular and wedge–shaped axes, pendants made of bear, pig, dog fangs, etc. animals, flint knife-shaped plates and arrowheads, shell beads, bird bones and copper tubes, copper pendants, bone punctures and pins, ceramics (from one to three vessels for burial) ^[2].

Fig. 1. The territory of Gavrilov-Yamsky district on the map of Yaroslavl county

         In the 1970s, according to A.L. Nikitin, another burial with a stone drilled axe and ceramics was destroyed during earthworks ^{[2, p. 57]}.

         The human skull, stored in the Archaeology Room of the P.G. Demidov YarSU, got there in the late 1970s. It was found during the development of a sand quarry on the site of the Nikultsinsky burial ground. One of the workers brought it to the university and handed it to I.L. Stankevich, who then headed the University archaeological expedition (YAGUAE). According to him, the bucket of the excavator hooked the burial. The rest of the bones and possible burial equipment were lost during the work. I.L. Stankevich went to the place of discovery, but no traces of burial could be found, because the place where the burial was already completely destroyed by the quarry. According to her, a small amount of Fatyanovo ceramics was found in the dump.

Figure 2. The village of Nikultsino on the ancient map of the Yaroslavl region.

DNA research data

A tooth belonging to the human remains described above was examined as a sample. The study of these samples was carried out under contracts with DNA-Heritage LLC no. DNA-SAS/01-21 dated 01.07.2021, no. DNA-La/02-22 dated 01.02.2022.

          According to the contracts, the determination of the STR loci of the Y chromosome by capillary electrophoresis of PCR products was ordered. The subsequent analysis and conclusions are copyrighted.

All stages of work with the archaeological sample were carried out in a fume hood located in a clean laboratory equipped with ULPA filters and UV lamps. In order to avoid contamination, all instruments and work space were sterilized in a clean laboratory using chemicals and harsh UV irradiation for 24 hours.

The tooth samples were cleaned from the top layer of dirt on special equipment. Further, the cleaned pieces of the tooth were kept under UV irradiation on each side for 30 minutes. Then the pieces of the tooth were crushed to the state of bone powder in a mill. As a result , dental powder weighing 0.5 g was obtained from the archaeological sample . For the sample, the DNA of the tooth powder was isolated by a method based on columns with SiO ₂.

 The concentration of the isolated DNA was evaluated on Qubit (HS) and amounted to 0.4 ng/ml. Using the Yfiler™ Plus PCR Amplification Kit Reagents(Thermo Fisher Scientific) fragment analysis of 27 STR loci of the Y chromosome was carried out on a capillary sequencer AB3500xl in two passes. To improve the accuracy of the result, two sequencing passes were performed. The results on the Y-chromosome haplotype of the samples were obtained in the IDX v.1.4 Gene Mapper program.

As a result, 18 STR loci of the Y chromosome were identified for the sample, indicated in Table. 1. Determination of subclades by loci through a predictor (https://www.nevgen.org ) gave the following results (Fig.5). Locus 518 gave a signal of a lower degree of clarity. 

       Fig. 3. A fore-graph of the result of the Y-STR fragments pollination. Passage 1.

Fig. 4. A fore-graph of the result of the Y-STR fragments pollination. Passage 2.

Table 1. Haplotype of the sample from Nikultino (names and alleles of STR loci)

  The sequencing data show that due to DNA degradation, 18 out of 27 possible loci were identified at two stages of sequencing, which makes it necessary to use a predictor program to assign a haplotype to a haplogroup. The predictor used nevgen.org showed the probability of attributing the haplotype to haplogroup R1a, close to 100%.

Haplogroup R1a appeared in the Upper Paleolithic era, presumably in Southern Siberia about 20-15 thousand years ago and for many millennia moved westward. Between 10 and 8 thousand years R1a arrived in the Balkans and Central Europe. The distribution area of happogroup R1a is from Iceland, Germany to India, Iran and Altai.

Haplogroup R1a is believed to have been the dominant haplogroup among the northern and eastern Indo-European peoples, which partly passed into the Indo-Iranian (R1a1-Z93), partly into the branches of Central and Eastern European peoples (R1a-Z280, R1a-L664, R1a-M458).

Most carriers of haplogroup R1a belong to the subclades R1a1-M417. And within this haplogroup, the majority belongs to the R1a-Z645 population, whose representatives were native speakers of the first Indo-European languages. The Fatyanovo culture is a variant of the Indo-European culture of the Early Bronze Age on the Russian Plain.

In ^[4] there is a result of sequencing 24 representatives of the Fatyanovo culture (III millennium BC), and 14 individuals from the territory of the Yaroslavl region. All male burials belong to haplogroup R1a. For some of them (3 from the Yaroslavl region) The subclades Z93, which is now characteristic of the Indo-Iranian peoples and cultures of the Great Steppe, were identified, but no haplotypes were given in the article. Six individuals belong to the Nikultsinsky burial ground (excavations by D. Krainov), for two of the three male burials, haplogroup R1a is indicated without detailing.  The interpretation of the presence of Z93 in the graves of the Yaroslavl region may be the possibility of penetration and further spread of R1a-Z93 carriers through the Upper Volga to the east, into the forest-steppe and steppe regions in the III-II thousand BC, into the region of the Sintashta culture.

An important circumstance for the interpretation of the haplotype is that those representatives of the Fatyanovo culture for whom the subclades R1a-Z93 (2900-2500 BC) were identified are not much distant in time from the very appearance of R1a-Z93^[5] (3000 BC). Therefore, in the case of this subclades, it must have basal, archaic forms not far removed from the basal forms of other related subclades (primarily, Z280). The paper ^[1] pointed out the importance of the Fatyan culture for understanding the conditions of occurrence and divergence of R1a-Z280, and the possible presence of both R1a-Z280 and R1a-Z93 in this culture. The predictor data show that the haplotype from Niculcino is sufficiently close in the most diverse variants of R1a-Z645. Moreover, the Z93 subclades have a serious probability of similarity. The proximity to different subclades at the same time can probably be explained by the "basal", ancestral character of the haplotype, its proximity to the basal form for R1a-Z645, R1a-Z283, R1a-Z93.

Figure 5. Data nevgen.com assignment of the haplotype to subclades R1a (March 2022).

However, there is still not enough data for a final conclusion, and based on the highly probable attribution of the haplotype to R1a-Z93 and the simultaneous proximity to other variants, we can only conclude that, based on DNA data, the hypothesis that the burial belongs to the Fatyanovo culture of early Bronze cannot be rejected.

Thanks

The team of authors thanks Gennady Tsvetkov, Vasily Savransky, Vladimir Krupnov, Mikhail Voinov, Margarita Komova, Natalia Lipatnikova, Elena Oleinikova, Olga Mokrushina, Olga Zaitseva, Andrey Semenov, Alexey Simonov for financial assistance in conducting the study.