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UNIVERSITATEA DE VEST TIMIŞOARA ARHEOVEST IV 2 -IN HONOREM ADRIAN BEJAN- Interdisciplinaritate în Arheologie și Istorie Timişoara, 26 noiembrie 2016 JATEPress Kiadó Szeged 2016

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  • UNIVERSITATEA DE VEST TIMIŞOARA

    ARHEOVEST

    IV2

    -IN HONOREM ADRIAN BEJAN-

    Interdisciplinaritate în Arheologie și Istorie

    Timişoara, 26 noiembrie 2016

    JATEPress Kiadó

    Szeged 2016

  • Coordonator volum: Dorel MICLE Editori: Dorel MICLE, Andrei STAVILĂ, Cristian OPREAN, Sorin FORȚIU Coperta: Alice DUMITRAȘCU Foto copertă: Milan ȘEPEȚAN Această lucrare a apărut sub egida:

    © Universitatea de Vest din Timișoara https://www.uvt.ro/

    ISBN 978-963-315-310-9 (Összes/General) ISBN 978-963-315-312-3 (II. kötet/volumul)

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    DVD-ROMul conține contribuțiile în varianta color precum și imaginile la rezoluția maximă trimisă de autor.

    SorinSticky NoteAvertisment Acest volum digital este o imagine cât se poate de fidelă a celui tipărit. Doar paginile albe din volumul tipărit au fost omise.

  • 631

    DNA ANALYSIS OF A NOUA CULTURE FUNERARY CONTEXT FROM EASTERN ROMANIA

    Neculai Bolohan*, Mitică Ciorpac**, Florica Mățău***, Dragoș-Lucian Gorgan****

    * Faculty of History, Alexandru Ioan Cuza University of Iași; [email protected] ** Interdisciplinary Research Department-Field Science, Alexandru Ioan Cuza University of Iași; [email protected] *** Interdisciplinary Research Department-Field Science, ARHEOINVEST Platform, Alexandru Ioan Cuza University of Iași; [email protected] **** Faculty of Biology, Alexandru Ioan Cuza University of Iași; [email protected] Rezumat. Acest studiu prezintă primele rezultate ADN mitocondrial vechi (amtDNA) obți-nute din analiza unor fragmente osteologice umane ce provin dintr-un context funerar atribuit Bronzului târziu care au avut ca scop identificarea haplogrupului precum și începutul consti-tuirii unei baze de date pentru ADN-ul mitochondrial vechi din estul României. Fragmentele osteologice provin de la un individ identificat în cadrul sitului arheologic de la Tăcuta (jud. Vaslui), care este localizat în partea central-nordică a Podișului Moldovenesc, în bazinul Dobro-vățului. Pe baza inventarului asociat, acest context funerar a fost atribuit culturii Noua din estul României.

    ADN-ul mitocondrial reprezintă un singur haplotip care este transmis de la mamă către urmași, ceea ce înseamnă că liniile mitocondriale pot fi identificate într-un mod mult mai sim-plu decât liniile nucleare. Extragerea ADN-ul mitocondrial din fragmentele osteologice implică două activități principale: prepararea probelor și protocolul de izolare. Pregătirea probelor a urmat protocolul standard pentru analize ADN, care implică utilizarea echipamentului de labo-rator (măști, mănuși, halate) într-un mediu steril și fără contaminare ADN/ARN.

    Analiza ADN-ului mitocondrial extras din fragmentele osteologice atribuite culturii Noua a permis identificarea coabitării, dar și a mobilității comunităților umane aparținând Bron-zului târziu din estul României. Cuvinte cheie: amtADN, haplogrup, context funerar, cultura Noua, estul României.

    1. Introduction The Noua culture represents the Late Bronze Age which developed during

    the second half of the 16th century and the 12th century BC in a large area, mainly, between the North and North-Western Black Sea and Western Carpathians region. The main cultural features are the ash-heap settlements and the very large number of funerary contexts, especially when compared to other LBA cultures. In terms of mate-riality (pottery, metal and bone artefacts) and funerary customs it is related to the Sabatinovka culture, which is spread in the northern part of the Black Sea region.

    SorinTypewritten Textreferințăbibliografică

    SorinSticky NoteNeculai Bolohan, Mitică Ciorpac, Florica Mățău, Dragoș-Lucian Gorgan, DNA analysis of a Noua Culture funerary context from Eastern Romania, În: ArheoVest, Nr. IV: In Honorem Adrian BEJAN, Interdisciplinaritate în Arheologie și Istorie, Timișoara, 26 noiembrie 2016, Vol. 1: Arheologie, Vol. 2: Metode Interdisciplinare și Istorie, Universitatea de Vest din Timișoara, JATEPress Kiadó, Szeged, 2016, Vol. 1: pp. 1–532 + DVD, Vol. 2: pp. 533–982, ISBN 978-963-315-310-9 (Összes/General), ISBN 978-963-315-311-6 (Kötet/Vol. 1), ISBN 978-963-315-312-3 (Kötet/Vol. 2); Vol. 2, pp. 631–638.

  • 632

    The Noua funerary practices included inhumations and cremations in mounds, intra-mounds and in flat burials, with a total of cca. 144 funerary contexts1.

    Fig. 1. The location of the funerary context from Tăcuta, Vaslui County, RO.

    Most of the burials are included in large cemeteries, but small groups of burials and isolated flat tombs exist, also. The funerary context (Tm7) identified at Tăcuta-Dealul Miclea/Paic (Vaslui County, RO) which consists in a flat burial was identified in a region reach in Noua settlements and funerary discoveries (Fig. 1).

    The osteological remains were not in a very good state of preservation (Fig. 2). The burial contains three vessels as grave goods which were attributed based on their typological characteristics to the Noua culture2.

    If we are interested in tracing the mobility of the LBA communities and/or individuals (when they moved? how many peoples moved? how many times they moved?) there is the need to get insights into the genetic data, mainly, the mitochon-drial DNA sequence. The DNA analysis will show us the patterns of variations in population and, then, we can compare this data with the patterns of variations in the materiality and funerary customs. In order to do this, we intend to create a database which will give us the possibility to connect our genetic approach and results with other European research programs which are targeting South-Eastern Europe.

    1 Motzoi-Chicideanu, 2011, p. 564-599. 2 Boghian et alii, 2013, p. 205-206.

  • 633

    Fig. 2. The funeray context from Tăcuta3.

    2. Methodology For a higher confidence degree, two extraction protocols were used: phenol-

    chlorophorm-isoamyl alcohol (PCI) and DNA IQ (Promega, USA). For both extrac-tion protocols blank control samples were used to assess the potential reagents or lab contamination. In order to identify any possible contamination that might have occurred in the different stages of the samples preparation and mainly in the aDNA isolation, at least two extraction blank controls and multiple PCR non-template controls were included in each amplification reaction.

    The mitochondrial hyper variable regions 1 and 2 (Hv1 and Hv2) were ampli-fied and direct sequenced.

    3 Boghian et alii, 2013.

  • 634

    3. Results The Hv1 and Hv2 sequences were concatenated analysed in BLAST module

    from the National Center for Biotechnology Information (NCBI). The comparison revealed that sample have a high similarity score with HV haplogroup.

    Sequence Predicted Haplogroup Total Variants Variants

    TM7 HV (HV0) 1 T16298C

    Table 1. Mitomaster DNA identity analysis.

    Human mitochondrial DNA can be divided in genealogical groups sharing a common maternal ancestor, called haplogroups, being used in population genetics to trace back the (pre)historical population movements. The HV haplogroup is consid-ered the most successful maternal lineage in Europe and the Near East (Fig. 3). Over half of the European population and between 25% and 40% of the Near Eastern pop-ulation descends from a single common female progenitor who lived between 25,000 – 40,000 years ago. This situation could also be associated with a repopulation event that began after the last glacial sequence. A recent phylogenetic reconstruction indi-cates that some lineages within haplogroup HV, which was previously included as a whole in the “mitochondrial Neolithic package” as a characteristic marker of the Linear Pottery culture in central Europe, might have left traces of an early diffusion also in the southern regions, without being able to clarify if this early diverging lineages would have arrived before or after the Neolithic diffusion4. This common ancestor is the found-er of the HV lineage, a characteristic of most Europeans as descends from a branch that was renamed haplogroup H. Another small but substantial European branch was called haplogroup V.

    Haplogroups Networks were constructed for H and HV haplogroups separately, and then they were combined to show the general topology. Reticulations were re-solved whenever possible by assuming all the back mutations in the less frequent haplotypes. Subclusters could be confidently identified in the HV1, HV2 network.

    The Tm7 amtDNA sequences have a high similarity with HV and HV0 clus-ters as shown by the network analysis (Fig. 4), which confirms the descendancy from a common ancestor.

    4 de Fanti et alii, 2015, p. e0144391.

  • 635

    Fig. 3. Identified HV haplogroup distribution in Europe

    and their relative frequency5.

    Fig. 4. Haplogroups network.

    5 http://www.eupedia.com/europe/Haplogroup_HV_mtDNA.shtml (accessed 15.10.2016).

  • 636

    Fig. 5. Evolutionary relationships of taxa.

    The evolutionary history presented in Fig. 5 was inferred using the Neighbor-Joining method6. The optimal tree with the sum of branch length = 0.09585898 is shown. The confidence probability (multiplied by 100) that the interior branch length is greater than 0, as estimated using the bootstrap test (500 replicates is shown next to the branches7. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Maximum Composite Likelihood method8and are in the units of the number of base substitutions per site. The analysis involved 23 nucleotide sequences. All positions containing gaps and missing data were eliminated. There were a total of 381 positions in the final dataset. Evolutionary analyses were conducted in MEGA69.

    6 Saitou, Nei, 1987, p. 406-425. 7 Dopazo, 1994, p. 300-304; Rzhetsky, Nei, 1992, p. 945-967. 8 Tamura et alii, 2004, p. 11030-11035. 9 Tamura et alii, 2013, p. 2725-2729.

  • 637

    4. Conclusions Genetic data for sample TM7 confirms previously analysed LBA samples

    from Romania which were assigned to Noua culture based on the funerary context and associated grave goods.

    The HV haplogroup on the present-day Romania territory was identified in samples belonging to Early Neolithic (Starčevo-Criș culture), Middle Neolithic (Zau culture) and in two other LBA samples selected from Transylvania10.

    Although some populations from LBA and Early and Middle Neolithic from Romania shared the same haplogroup, previous investigations concluded based on multidimensional scaling analysis that the LBA samples shows genetic similarity with the populations from Ukraine.

    Acknowledgements The financial support for this study was provided by the the PCCA 1153/2011

    Nr. 227/01.10.2012 Genetic Evolution: New Evidences for the Study of Interconnected Structures. A Biomolecular Journey around the Carpathians from Ancient to Medieval Times (GENESIS).

    We would like to express our gratitude to the members of the Tăcuta-Dealul Miclea/Paic archaeological team (Dumitru BOGHIAN, Sergiu-Constantin ENEA, Ciprian-Cătălin LAZANU, Andrei ASĂNDULESEI, Felix-Adrian TENCARIU) who provided the osteological samples from the excavation held in 2012.

    10 Hervella et alii, 2015, p. e0128810.

  • 638

    BIBLIOGRAPHY Boghian et alii, 2013

    Dumitru BOGHIAN, Sergiu-Constantin ENEA, Felix TENCARIU, Dia-na-Măriuca VORNICU, Andreea VORNICU, Loredana PRICOP, Ale-xandru GAFINCU, Radu-Gabriel FURNICĂ, Bogdan MUNTEANU, Tăcuta, com. Tăcuta, jud. Vaslui, In: Cronica Cercetărilor Arheolo-gice din România. Campania 2012, A XLVII-a Sesiune Națională de Rapoarte Arheologice, Craiova, 27‒30 mai 2013, Ministerul Culturii, Institutul Național al Patrimoniului, Comisia Naţională de Arheologie, Direcţia Patrimoniu Cultural, Muzeul Olteniei - Craiova, Ed. Universității “Alexandru Ioan Cuza”, Iași, 2013, 385 pg., ISSN 23-43-919X; ISSN-L 23-43-919X; p. 205-206.

    Motzoi-Chicideanu, 2011

    Ion MOTZOI-CHICIDEANU, Obiceiuri funerare în epoca bronzului la Dunărea Mijlocie și Inferioară, Vol. I, Ed. Academiei Române, Bucu-rești, 2011, 900 pg., ISBN 9732720468, 9789732720462.

    Dopazo, 1994 Joaquin DOPAZO, Estimating errors and confidence intervals for branch lengths in phylogenetic trees by a bootstrap approach, In: Journal of Molecular Evolution, 38, 1994, p. 300-304.

    de Fanti et alii, 2015

    Sara de FANTI, Chiara BARBIERI, Stefania SARNO, Federica SEVINI, Dario VIANELLO, Erika TAMM, Ene METSPALU, Mannis van OVEN, Alexander HÜBNER, Marco SAZZINI, Claudio FRANCESCHI, Davide PETTENER, Donata LUISELLI, Fine Dissection of Human Mitochon-drial DNA Haplogroup HV Lineages Reveals Paleolithic Signatures from European Glacial Refugia, In: PloS ONE, 10 (12), 2015, e0144391.

    Hervella et alii, 2015

    Montserrat HERVELLA, Mihai ROTEA, Neskuts IZAGIRRE, Mihai CONSTANTINESCU, Santos ALONSO, Mihai IOANA, Cătălin LAZĂR, Florin RIDICHE, Andrei Dorian SOFICARU, Mihai G. NETEA, Con-cepcion de-la-RUA, Ancient DNA from South-East Europe Reveals Different Events during Early and Middle Neolithic Influencing the European Genetic Heritage, In: PloS ONE, 10 (6), 2015, p. e0128810.

    Rzhetsky, Nei, 1992

    Andrey RZHETSKY, Masatoshi NEI, A Simple Method for Estimating and Testing Minimum-Evolution Trees, In: Molecular Biology and Evo-lution, 9, 1992, p. 945–967.

    Saitou, Nei, 1987

    Naruya SAITOU, Masatoshi NEI, The neighbor-joining method: a new method for reconstructing phylogenetic trees, In: Molecular Biology and Evolution, 4, 1987, p. 406-425.

    Tamura et alii, 2004

    Koichiro TAMURA, Masatoshi NEI, Sudhir KUMAR, Prospects for inferring very large phylogenies by using the neighbor-joining method, In: Proceedings of the National Academy of Sciences of the United States of America, 101, 2004, p. 11030-11035.

    Tamura et alii, 2013

    Koichiro TAMURA, Glen STECHER, Daniel PETERSON, Alan FILIPSKI, Sudhir KUMAR, MEGA6: Molecular Evolutionary Genetics Analysis version 6.0., In: Molecular Biology and Evolution, 30, 2013, p. 2725-2729.