1. Introduction
The present work will shed some light on the ancestry and physical appearance of Princess Leonor of Castile. To this commitment, we have applied ancient DNA techniques. We understand ancient DNA, which has a low quantity and quality of DNA molecules due to its significant age. Not always antiquity is the main cause of DNA degradation, but when we face up to any potential critical sample, like in this case, it must be implemented special analysis conditions and authenticity criteria (Gomes et al. 2019).
In a colloquium about kinship and legitimation, the Portuguese historian J. Mattoso (2011) referred to the importance of the Medieval research of words and language to understand the modalities and evolution of discourse, since those voices would be “a bit like mitochondrial DNA and chromosome Y, which is transmitted from generation to generation, and therefore, always keep some vestiges of previous generations”. He was referring to the fact that these genetic lineage markers are transmitted unaltered from generation to generation to trace people’s genealogy.
Beyond the fortunate metaphor of Mattoso, in the field of Medieval and Modern History, the application of Genetics has also changed, with the DNA analysis of individuals found in the necropolis, but also, for obvious reasons, of sets of remains from royal pantheons, for example from Aragon (Martínez Jarreta 2018) or Hungary (Olasz et al. 2019). Works oriented to the identification of controversial characters are already frequent (Salerno et al. 2005; Haeusler et al. 2016) or to establish kinship among individuals buried together (Gomes et al. 2020; Gamba et al. 2011), without neglecting the detection of individuals related to skeletons with signs of serious diseases (Alves-Cardoso et al. 2022). Nowadays, it is even possible to determine the physical appearance of individuals by the genetic analysis of their skeletal remains (Gomes et al. 2017, 2020). Today it can be said, as Larmuseau and Bodner (2018) have pointed out, that genealogical genetic analyses have, above the merely anecdotal, significative biological, historical, social, and educational importance.
On this occasion, we present the study carried out on the remains of the Infanta Leonor, daughter of King Alfonso X the Wise, as a part of a project for the conservation, restoration, and dissemination of her remains and sepulcher.
Leonor of Castile, an Unknown Princess
The Spanish Princess Leonor of Castile (1256/1257–1275) was the fourth daughter of eleven legitimate sons and daughters of Alfonso X “The Wise” (Spain, 1221–1284) and the Queen Violante of Aragón (1236–1300) (de Salazar Acha 1990). On one hand, there is certain conflicting information about the life of Princess Leonor from different sources (González González and Iriarte 1993; Gutierrez Baños 2014). On the other hand, no registered graphical documents about her physical appearance have survived our days.
Concerning her biogeographical origin, the genealogy of her family is summarized in Figure 1. In summary, in this figure (Figure 1), it is possible to observe that, according to the historical sources, princess Leonor of Castile was fundamentally European and to some extent with Eastern Mediterranean ancestry.
However, it is interesting to analyze in more depth the maternal lineage of Lady Leonor due to the exclusively maternal lineage inheritance of one of the genetic markers studied in this work, mitochondrial DNA (mtDNA). Most of the maternal lineage of Lady Leonor is summarized in Figure 2.
The princess died when she was 19 years old of unknown causes (Martínez Santamarta and Robert 2010). She probably died in Montpellier (France); however, Alfonso X ordered that Leonor’s remains be buried in the Royal Monastery of Saint Dominic of Caleruega (Burgos, Spain), where they were located up to the beginning of this research (Gutierrez Baños 2014).
The present study revolves around two objectives: on one hand, to determine the biogeographical origin of the remains to prove if the predicted biogeographical origin is consistent with the supposed European Princess’ ancestry; and, on the other hand, to determine the most likely pigmentation phenotype (skin, eyes, and hair pigmentation) of Lady Leonor of Castile.
2. Materials and Methods
On June the 10th, 2014, the opening of the sarcophagus containing the mummified remains of Lady Leonor of Castile was carried out. Previously, the coffin had been subjected to a Helical Computed tomography and EDX analysis (Energy Dispersive X-Ray Analysis, an x-ray technique used to identify the elemental composition of materials) (Pastor et al. 2021). Subsequently, the coffin was opened and the remains found wrapped in a piece of white fabric. Through the first stage of the research, the clothing and other objects were removed to be analyzed and reconstructed.
To perform the genetic analysis, there were selected two well-preserved teeth: the lower left second premolar 1LDC1 and the lower left first molar 1LDC2 (Figure 3).
Genetic analyses were performed in two specialized aDNA laboratories, hereinafter, Laboratory 1 (at Health Legislation, Psychiatry, and Pathology Department. Medicine Faculty. Complutense University of Madrid) and Laboratory 2 (at Medicine and Odontology Faculty, Santiago de Compostela University); both yearly tested and certified by GHEP-ISFG (Spanish and Portuguese speaking group of the International Society of Forensic Genetics).
DNA extraction was performed in Laboratory 1 following Gomes et al. (2015). Following this method, sample 1LDC1 and sample LDC2 were analyzed in separate processes. Sample 1LDC1 was pulverized, and the DNA from 1LDC2 was extracted without the physical destruction of the piece. And hereinafter, both sample DNA extracts were processed separately to replicate results.
-
(a). The amplification of the DNA regions of interest supplied us with two different kinds of information: biogeographical origin and externally visible characteristics (EVCs).
Biogeographical origin analysis: To study the individuals’ most probable origin, it was taken into account two kinds of information: nuclear and mitochondrial DNA (mtDNA). To predict the most probable biogeographic origin with nuclear information, the SNPforID 34-Plex forensic ancestry test was used to analyze 34 SNPs (Phillips et al. 2007; Fondevila et al. 2013). The PCR conditions are collected in Table 1. After the analysis in an ABI 3500 Genetic Analyzer (ThermoFisher SCIENTIFIC), biogeographic SNPs allele information was analyzed using GeneMapper™ Software 5 (ThermoFisher SCIENTIFIC, Waltham, MA, USA).
Finally, statistical parameters and the most probable biogeographical region were obtained with the software The Snipper app. suite v.2.5 (Phillips et al. 2007), considering the dataset for three major populations: Europe, Africa, and East Asia. This analysis was performed in Laboratory 2.
To analyze the maternal origin by mtDNA and determine the most probable haplogroup(s), both HV1 and HV2 regions were analyzed in Laboratory 1.
There were performed the amplification of 294 bp of the Hypervariable region I (HVRI) (positions 16,105–16,399) (Fernández 2005) and 345 bp of the Hypervariable region II (HVRII) (positions 55–400) (Martínez-Labarga and Rickards 1999). These regions were selected and incorporated into four overlapping PCRs using the Multiplex PCR kit of Qiagen® (Hilden, Germany). The analysis was performed in duplicate for each of the selected samples. The amplification results were tested by electrophoresis on agarose gels and positive amplification results were purified with ExoSAP-IT™ PCR Product Cleanup Reagent (Thermo Fisher Scientific, Waltham, MA, USA) (Exo-SAP®). Sequencing products were visualized on an ABI3730 (Life Technologies: Carlsbad, CA, USA) automated capillary electrophoresis sequencer.
The capillary electrophoresis results were analyzed with the aid of the software Chromas® (
-
(b). For the external phenotypic prediction, 35 SNPs were analyzed in Lab 2 to calculate the most probable hair, skin, and eye color pigmentation, using the primers and PCR conditions of Ruiz et al. (2013) (Maroñas et al. 2014). The EVCs PCR conditions are collected in Table 2. For each externally visible characteristic (EVC), the hypotheses considered were (a) iris pigmentation (brown, intermediate, and blue); (b) hair pigmentation (fair versus dark, and for the color pigmentation (red, blond, brown, and black); and (c) skin pigmentation (white, intermediate, and black) (Maroñas et al. 2014). Statistical analysis was performed with the online Snipper app suite v2.5 software (Phillips et al. 2007).
Along with the full analytical procedure, pre-PCR, PCR, and post-PCR procedures were performed in three physically separated and isolated areas. Ancient DNA laboratories were equipped with UV light lamps (254 nm) to ensure a sterile environment for the reliable analysis of ancient DNA samples. Additionally, all laboratory equipment was routinely irradiated with UV light and thoroughly cleaned with bleach (70%) before and after each experiment. All experimental analyses were conducted by a single researcher to reduce the chances of staff DNA contamination; additionally, the access to these laboratories was limited to two people during the whole time the analyses were carried out. All laboratory procedures (sample preparation, extraction, and PCR) were performed wearing disposable laboratory coveralls (masks, caps, glasses, shoe covers, and gloves). DNAse and RNase-free reagents and consumables were also employed. The possibility of modern DNA contamination was monitored with extraction blanks and at least three PCR-negative controls were included per DNA amplification round. So, authenticity criteria were followed, considering the laboratories’ infrastructures, methodology, and interpretation of the results. Also, before the genetic analysis, two biochemical assays on macromolecular preservation were performed by the archaeological team, as recommended by Pääbo (Pääbo et al. 2004). To monitor exogenous mitochondrial DNA contamination of the samples, genetic profiles were recovered from all the people involved in sample manipulation (Table 3).
3. Results
At Laboratory 1, all the PCR reactions performed have provided quality mtDNA sequences without molecular damage or trace of contamination. Furthermore, all the obtained results were consistent among them. This fact provides further strength to the obtained results. Combining the different PCR amplifications has allowed us to determine the next consensus mtDNA haplotype: 16224C 16311C 73G 195C 263G 315.1C (range: 16,105–16,569; 1–390). This result was obtained at least twice from each sample and mtDNA fragment, always with the same result. Subsequently, the obtained results were analyzed to determine the mitochondrial haplogroup, and it was determined as K1a4a1a, with a private mutation on position number 195 of HRVII.
Concerning the amplification of 34 autosomal SNPs for biogeographical assignment carried out at Laboratory 2, results can be seen in Table 4. This table also shows the obtained LR results performed to discern if the individual belongs to a European, Sub-Saharan Africa, East of Asia, American or Oceanian population.
The autosomal SNPs analysis suggests a European origin in the same way as the mtDNA analysis.
Finally, the results of the EVC genetic analysis carried out in Laboratory 2 are summarized in Table 5. To determine each one of the physical characteristics studied, we performed the same methodology employed in the biogeographical origin analysis.
In light of these results, the most probably physical appearance of the Princes was black hair, green-hazel eyes, and white skin.
4. Discussion
We have determined that the human remains studied belong to mtDNA haplogroup K1a4a1a, a typical European haplogroup nowadays, this finding indicates a typical European or Middle East biogeographical ancestry; this is consistent with the European maternal lineage origin of the Princess. As regards ancient DNA, the earliest reported appearance of this haplogroup was dated around 8000 years B.P. in Tell Ramad (Siria) (Fernandez et al. 2014). Also, human remains belonging to haplogroup K have been discovered, which date around 7500–7300 B.P. One of the most popular findings was Ötzi (5000 B.P.), which precisely belongs to the K1 haplogroup (Rollo et al. 2006). Through searching on the AmtDB database of the K1a4a1a haplogroup, no individuals were found with the same haplogroup. Nevertheless, also in AmtDB, the closest haplogroup found in Medieval populations was located in Hungary (Vai et al. 2019). K1a clade (group of individuals who share the same lineage, K1a) was also present during Middle Ages in Germany (Veeramah et al. 2018). If we search the broader K1, it was also present during Middle Ages in Italy (Amorim et al. 2018; Vai et al. 2019), Spain (Olalde et al. 2019), and Poland (Stolarek et al. 2018); but the largest presence of K1 mtDNA haplogroup was located in Hungary. These results are consistent with the mtDNA family tree of Leonor Princess (Figure 2).
Moreover, autosomal SNPs analysis supports this European origin too. This second analysis furthermore provides a broader view of the Princess’ origins, since the analyzed SNPs have been inherited through both the maternal and the paternal lineage and are consistent too with the historical genealogy of Lady Leonor´s family, with mainly European roots (Figure 1 and Figure 2). The matching results for the prediction of biogeographical origin obtained through the two different methodologies employed, provide greater strength to the concluded ancestry.
It could be interesting to compare Lady Leonor´s mtDNA haplotype to that of any person belonging to this maternal genealogy tree. However, it has not been possible to find any previously generated genetic data from other individuals from this maternal lineage. However, the mtDNA and YSTRs profiles of her grandmother´s grandfather, the Hungarian king Bela III, is well known (Olasz et al. 2019); unfortunately, this information is not comparable to our results. Nevertheless, the newly obtained data can provide relevant information to complete the genetic familiar genealogy.
Moreover, the feature that genetic evidence supports as most likely white skin pigmentation seems also to support a possible European origin for the studied remains.
From the paleogenetic point of view, this work confirms the feasibility of genetics to establish the kinship and physiognomic features of individuals. Determining that she is most likely to have white skin, green-hazel eye color, and black hair. Thereby, the Princess obscured in the documentation acquires some visibility. In addition, its haplogroup, K1a4a1a, has been obtained, establishing an eventual link with other royal houses currently under investigation (Olasz et al. 2019).
Certainly, from a historical point of view, the knowledge of notable characters is not today one of the main objectives of investigations, but the realization of restoration and conservation works constitutes new potential sources, usable from different disciplines and prospects. In this sense, the present study has an experimental nature, of exploration possibilities, with which some bases are laid whose use will be greater when other similar ones that allow comparative approaches are published.
Working with these royalties sometimes allows for written descriptions of their physical appearance and sometimes even preserved remains—such as the nails of the blonde Norwegian princess Kristín Hákonardóttir, during some time Leonor’s mother’s rival, whose tomb was opened last century (Vargas Blanco 1968), and sometimes there are also some “portraits”, such as the miniatures of the monarchs, queens or princess collected in the cardboards of the cathedrals from Oviedo and Compostela, which confront genetic inferences, but, as has been said, none of Lady Leonor seems to be known until now. In general, the possibilities of identification will be greater from the Late Middle Ages, with the progressive display of the pictorial portrait.
The viability of the comparisons also grows in the case of investigations such as those mentioned above of the royal pantheons of Hungary and Aragon. Although there were partial pantheons in San Isidoro (León, Spain) and Las Huelgas (Burgos, Spain), for complex historical reasons (Arias Guillén 2015) a single royal pantheon was not formed in the whole of the Crown of Castile that could have facilitated this type of genetic research, it is not impossible to carry it out, taking advantage—as in the present case—projects of rehabilitation of scattered graves. The location of the characters of the dynasties—for example, Leonor’s maternal grandmother and transmitter of her mitochondrial DNA, Violante of Hungary, Aragonese Queen buried in the Royal Monastery of Santa María de Vallbona (Lérida, Spain)—may serve for the constitution of a genetic database. In this sense, the genetic analysis of Leonor of Castile comes to join those already available for some of his relatives, such as Bela III of Hungary and Agnes de Châtillon, grandparents precisely from the aforementioned maternal grandmother of Leonor (Olasz et al. 2019).
Conceptualization, J.F.P., E.A.-P., C.G. and S.P.-D.; methodology, S.P.-D., C.G. and M.S.F.; software, S.P.-D., C.G. and M.S.F.; validation, S.P.-D., C.G., M.S.F. and A.M.L.-P.; formal analysis, S.P.-D. and C.G.; investigation, S.P.-D., C.G.; resources, S.P.-D., Á.E.-A., J.F.P.; data curation, S.P.-D., C.G., M.S.F. and A.M.L.-P.; writing—original draft preparation, S.P.-D.; writing—review and editing, S.P.-D., C.G., M.S.F., Á.E.-A., A.M.L.-P., M.V.L., E.A.-P. and J.F.P.; visualization, S.P.-D., C.G., M.S.F., Á.E.-A., A.M.L.-P., M.V.L., E.A.-P. and J.F.P.; supervision, S.P.-D., C.G., M.S.F., and E.A.-P.; project administration, J.F.P.; funding acquisition, J.F.P. All authors have read and agreed to the published version of the manuscript.
Not applicable.
Not applicable.
All available data is in this article.
We declare that the authors have no competing interest as defined by MDPI, or other interest that might be perceived to influence the results and/or discussion reported in this paper.
Footnotes
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.
Enlarge this image.Figure 2. Maternal lineage Biological tree of Lady Eleanor of Castile. The scheme shows all the family members within Princess Eleanor’s maternal lineage with whom they share the same mitochondrial DNA.
The SNPforID 34-Plex PCR conditions.
| PCR Conditions | Cycling Protocol | ||||
|---|---|---|---|---|---|
| Final Concentration | Volume (mL) | ||||
| Buffer | 1 X | 0.615 | |||
| BSA | 3.2 mg/mL | 0.615 | 95 °C | 15 min | |
| Cl2Mg | 6.3 mM | 1.615 | 95 °C | 30 s | 35 cycles |
| dNTPs | 0.625 mM | 0.4 | 60 °C | 50 s | |
| Primer | Variable | 2 | 65 °C | 40 s | |
| TaqGold | 0.5 U | 0.1 | 65 °C | 6 min | |
| DNA | n/a | 1–4 mL | |||
EVCs PCR conditions.
| PCR Conditions | Cycling Protocol | ||||
|---|---|---|---|---|---|
| Final Concentration | Volume (mL) | ||||
| buffer | 1 X | 0.625 | 95 °C | 15 min | |
| BSA | 3.2 mg/mL | 0.625 | |||
| Cl2Mg | 6.3 mM | 1.625 | 95 °C | 30 s | 35 cycles |
| dNTPs | 0.625 mM | 0.43 | 60 °C | 50 s | |
| primer | Variable | 1.5 | 65 °C | 40 s | |
| TaqGold | 0.5 U | 0.1 | |||
| DNA | n/a | 1–4 uL | 65 °C | 6 min | |
Researchers’ mitochondrial DNA profiles.
| Laboratory Staff | Mitochondrial DNA Haplotype |
|---|---|
| RESEARCHER A | 16362C 55G 150T 239C 263G 309.1C 309.2C 315.1C |
| RESEARCHER B | 263G 315.1C |
| RESEARCHER C | 16145A 16172C 16222T 16261T 16305T 73G 242T 263G 295T 309.1C 315.1C |
| RESEARCHER D | 263G 315.1C |
| RESEARCHER E | 16126C 16294T 16296T 16304C 73G 263G 315.1C |
| RESEARCHER F | 73G 263G 315.1C |
Autosomal SNPs results for the Biogeographical origin analysis and Likelihood Ratio results of the statistical comparison of the alleged origins one against another. In columns, we can observe the SNP (single nucleotide polymorphisms) analyzed and the obtained results obtained for each one. At the bottom of the table, it is possible to view the Likelihood ratio (LR) results obtained to discern if the individual belongs to a European, Sub-Saharan Africa, East of Asia, American or Oceanian population, showing that the most probable biogeographical origin is European, attending to the different origin hypothesis.
| SNP | RESULT | SNP | RESULT | SNP | RESULT |
|---|---|---|---|---|---|
| rs5997008 | C | rs773658 | C | rs2026721 | G |
| rs2304925 | GT | rs10141763 | A | rs4540055 | A |
| rs917118 | G | rs182549 | CT | rs1335873 | A |
| rs1321333 | T | rs1573020 | A | rs16891982 | C |
| rs2814778 | C | rs896788 | C | rs730570 | T |
| rs1024116 | A | rs2065160 | A | rs1886510 | NN |
| rs7897550 | C | rs2572307 | G | rs5030240 | C |
| rs722098 | AG | rs2303798 | C | rs3827760 | A |
| rs10843344 | CT | rs2065982 | A | ||
| rs12913832 | AG | rs3785181 | C | ||
| rs239031 | NN | rs881929 | NN | ||
| rs2040411 | AG | rs1498444 | A | ||
| rs1978806 | T | rs1426654 | T | ||
| LIKELIHOOD RATIO RESULTS | |||||
| HYPOTHESIS | LR | ||||
| H1: European origin vs. H2: American origin | LR = 1.95 × 1016 vs. 1 | ||||
| H1: European origin vs. H2: Asia origin | LR = 1.02 × 1018 vs. 1 | ||||
| H1: European origin vs. H2: North-African origin | LR = 10382,18 vs. 1 | ||||
| H1: European origin vs. H2: Sub-Saharian origin | LR = 2.18 × 1031 vs. 1 | ||||
EVCs SNPs results and Likelihood Ratio results of the statistical comparison of the EVCs one against another. In columns, we can observe the SNP (single nucleotide polymorphisms) analyzed and the obtained results for each. At the bottom of the table, it is possible to view the Likelihood ratio (LR) results obtained to discern the different external visible characteristics (EVCs) studied: eyes, skin, and hair colors.
| Iris Eye Color System | Hair Color System | Skyn Color System | |||
|---|---|---|---|---|---|
| SNP | RESULT | SNP | RESULT | SNP | RESULT |
| rs12913832 | AG | rs1129038 | NN | rs10777129 | NN |
| rs1129038 | NN | rs11547464 | NN | rs13289 | C |
| rs11636232 | CT | rs12913832 | AG | rs1408799 | G |
| rs12203592 | C | rs12931267 | C | rs1426654 | A |
| rs12896399 | A | rs1805006 | C | rs1448484 | A |
| rs1393350 | G | rs1805007 | NN | rs16891982 | C |
| rs1667394 | CT | rs1805008 | G | rs2402130 | A |
| rs16891982 | C | rs1805009 | G | rs3829241 | C |
| rs1800407 | C | rs28777 | NN | rs6058017 | NN |
| rs4778232 | G | rs35264875 | A | rs6119471 | C |
| rs4778241 | GT | rs4778138 | CT | ||
| rs7183877 | A | rs7495174 | CT | ||
| rs8024968 | G | ||||
| LIKELIHOOD RATIO RESULTS | |||||
| HYPOTHESIS | LR | HYPOTHESIS | LR | HYPOTHESIS | LR |
| H1: Dark hair vs. H2: Light hair | LR = 487.06 vs. 1 | H1: Green-hazel eyes vs. | LR = 26.71 vs. 1 | H1: White skin vs. | LR = 7.54 × 105 vs. 1 |
| H2: Brown eyes | H2: Intermediate skin | ||||
| H1: Black hair vs. H2: Brown hair | LR = 2.65 vs. 1 | H1: Green-hazel eyes vs. | LR = 2.64 × 106 vs. 1 | H1: White skin vs. | LR = 1.98 × 1010 vs. 1 |
| H2: Blue eyes | H2: Black skin | ||||
| H1: Black hair vs. H2: Red hair | LR = 215.63 vs. 1 | ||||
References
Alves-Cardoso, Francisca; Palomo-Díez, Sara; Alonso-Conde, Alejandro; Gomes, Cláudia; Casimiro, Silvia; Silva, Rodrigo Banha da; Arroyo-Pardo, Eduardo. Metagenomic Research of Infectious Diseases in Archealogical Contexts: Evidence from the Hospital Real de Todos-os-Santos (Portugal). Applied Sciences; 2022; 12, 6069. [DOI: https://dx.doi.org/10.3390/app12126096]
Amorim, Carlos Eduardo G.; Vai, Stefania; Posth, Cosimo; Modi, Alessandra; Koncz, István; Hakenbeck, Susanne; La Rocca, Maria Cristina; Mende, Balazs; Bobo, Dean; Pohl, Walter et al. Understanding the 6th-century barbarian social organization and migration through paleogenomics. Nature Communications; 2018; 9, pp. 1-11. [DOI: https://dx.doi.org/10.1038/s41467-018-06024-4] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/30206220]
Anderson, S.; Bankier, Alan T.; Barrell, B. G.; De Bruijn, M. H.; Coulson, Alan R.; Drouin, Jacques; Eperon, Ian C.; Nierlich, D. P.; Roe, B. A.; Sanger, Fred et al. Sequence and organization of the human mitochondrial genome. Nature; 1981; 290, pp. 457-65. [DOI: https://dx.doi.org/10.1038/290457a0]
Andrews, Richard M.; Kubacka, Iwona; Chinnery, Patrick F.; Lightowlers, Robert N.; Turnbull, Douglass M.; Howell, Neil. Reanalysis and revisión of the Cambridge reference sequence for human mitochondrial DNA. Nature Genetics; 1999; 23, 147. [DOI: https://dx.doi.org/10.1038/13779]
Arias Guillén, Fernando. Enterramientos regios en Castilla y León (c. 842–1504). La dispersión de los espacios funerarios y el fracaso de la memoria dinástica. Anuario de Estudios Medievales; 2015; 45, pp. 643-75. [DOI: https://dx.doi.org/10.3989/aem.2015.45.2.01]
de Salazar Acha, Jaime. «Precisiones y nuevos datos sobre el entorno familiar de Alfonso X el Sabio, fundador de Ciudad Real». Cuaderno de estudios manchegos; 1990; 20, pp. 211-31.
Fernández, Eva. Polimorfismos de ADN mitocondrial en poblaciones antiguas de la cuenca mediterránea. Ph.D. thesis; Universidad Central de Barcelona: Barcelona, Spain, 2005.
Fernandez, Eva; Pérez-Pérez, Alejandro; Gamba, Cristina; Prats, Eva; Cuesta, Pedro; Anfruns, Josep; Molist, Miquel; Arroyo-Pardo, Eduardo; Turbón, Daniel. Ancient DNA analysis of 8000 B.C. near eastern farmers supports an early neolithic pioneer maritime colonization of Mainland Europe through Cyprus and the Aegean Islands. PLoS Genetics; 2014; 10, e1004401. [DOI: https://dx.doi.org/10.1371/journal.pgen.1004401]
Fondevila, Manuel; Phillips, Christopher; Santos, Cristina; Aradas, Ana Freire; Vallone, P. M.; Butler, John M.; Lareu, María Victoria; Carracedo, Ángel. Revision of the SNPforID 34-plex forensic ancestry test: Assay enhancements, standard reference sample genotypes and extended population studies. Forensic Science International: Genetics; 2013; 7, pp. 63-74. [DOI: https://dx.doi.org/10.1016/j.fsigen.2012.06.007]
Gamba, Cristina; Fernandez, Eva; Tirado, Mirian; Pastor, Francisco; Arroyo-Pardo, Eduardo. Ancient Nuclear DNA and Kinship Analysis: The Case of a Medieval Burial in San Esteban Church in Cuellar (Segovia, Central Spain). American Journal of Physical Anthropology; 2011; 144, pp. 485-91. [DOI: https://dx.doi.org/10.1002/ajpa.21451]
Gomes, Cláudia; Remolins, Gerard; López-Parra, Ana María; Gibaja, Juan Francisco; Fondeliva, M.; De Angelis, Francesco; Veltre, V.; Subirá, María Eulália; Baeza-Richer, Carlos; Guerrero, D. et al. Paleogenetic evidence of a Pyrenean Neolithic family: Kinship, Physical appearance, and biogeography multidisciplinary analysis. Journal of Archaeological Science; 2020; 123, 105226. [DOI: https://dx.doi.org/10.1016/j.jas.2020.105226]
Gomes, Cláudia; Fondevila, Manuel; Magaña-Loarte, Concepción; Fernández-Jiménez, J.; Fernández-Serrano, José; Palomo-Díez, Sara; Baeza-Richer, Carlos; López-Parra, Ana María; Arroyo-Pardo, Eduardo. An unusual kinship case from the Spanish Civil War (1936–1939): Ancient versus degraded sample’s investigation. Forensic Science International: Genetics Supplement Series; 2019; 7, pp. 690-91. [DOI: https://dx.doi.org/10.1016/j.fsigss.2019.10.140]
Gomes, Cláudia; Fondevila, Manuel; Palomo-Díez, Sara; Pardiñas, Antonio F.; López-Matayoshi, César; Baeza-Richer, Carlos; López-Parra, Ana. María; Lareu, María Victoria; López, B.; Arroyo-Pardo, Eduardo. Phenotyping the ancient world: The physical appearance and ancestry of very degraded samples form a chalcolithic human remains. Forensic Science International: Supplement Series; 2017; 6, pp. e484-e486. [DOI: https://dx.doi.org/10.1016/j.fsigss.2017.09.188]
Gomes, Cláudia; Palomo-Díez, Sara; Roig, Joan; López-Parra, Ana; Baeza-Richer, Carlos; Espara-Arroyo, Angel; Gibaja, Juan; Arroyo-Pardo, Eduardo. Nondestructive extraction DNA method from bones or teeth, true or false?. Forensic Science International: Genetics Supplement Series; 2015; 5, pp. e279-e282. [DOI: https://dx.doi.org/10.1016/j.fsigss.2015.09.111]
González González, Carmen; Iriarte, Cándido Aniz. Real Monasterio de Santo Domingo de Caleruega; 1st ed. Editorial San Esteban: Salamanca, 1993; ISBN 978-84-87557-56-9
Gutierrez Baños, Fernando. El sepulcro de la infanta doña Leonor, hija de Alfonso X el Sabio, en el Real Monasterio de Santo Domingo de Caleruega (Burgos); Publicaciones de la Universidad de Santiago de Compostela. Quintana No. 13 2014 Universidad de Santiago de Compostela: Santiago, 2014; pp. 185-97.
Haeusler, Martin; Haas, Cordula; Lösch, Sandra; Moghaddam, Negahnaz; Villa, Igor M.; Walsh, Susan; Kayser, Manfred; Seiler, Roger; Ruehli, Frank; Janosa, Manuel et al. Multidisciplinary Identification of the Controversial Freedom Fighter Jörg Jenatsch, Assassinated 1639 in Chur, Switzerland. PLoS ONE; 2016; 11, e0168014. [DOI: https://dx.doi.org/10.1371/journal.pone.0168014]
Huber, Nicole; Parson, Walter; Dür, Arne. Next generation database search algorithm for forensic mitogenome analyses. Forensic Science International: Genetics; 2018; 37, pp. 204-14. [DOI: https://dx.doi.org/10.1016/j.fsigen.2018.09.001] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/30241075]
Larmuseau, Maarten H. D.; Bodner, Martin. The biological relevance of a medieval king’s DNA. Biochemical Society Transactions; 2018; 46, pp. 1013-20. [DOI: https://dx.doi.org/10.1042/BST20170173]
Lee, Hwan Young; Song, Injee; Ha, Eunho; Cho, Sung-Bae; Yang, Woo Ick; Shin, Kyoung-Jin. mtDNAmanager: A Web-based tool for the management and quality analysis of mitochondrial DNA control-region sequences. BMC Bioinformatics; 2008; 9, 483. [DOI: https://dx.doi.org/10.1186/1471-2105-9-483]
Maroñas, Olalla; Phillips, Chris; Söchtig, Jens; Gomez-Tato, Antonio; Cruz, Raquel; Alvarez-Dios, José; de Cal, María Casares; Ruiz, Yarimar; Fondevila, Manuel; Carracedo, Ángel et al. Development of a forensic skin colour predictive test. Forensic Science International: Genetics; 2014; 13, pp. 34-44. [DOI: https://dx.doi.org/10.1016/j.fsigen.2014.06.017] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/25082135]
Martínez Jarreta, María Begoña. Los estudios antropológicos de los restos óseos conservados. En Panteones Reales de Aragón; Gobierno de Aragón: Zaragoza, 2018; pp. 240-53.
Martínez Santamarta, H. Salvador; Robert, J. Alfonso X, the Learned: A biography. Volumen 146 de Studies in the History of Christian Traditions; 1st ed. BRILL: Leiden, 2010; ISBN 9789004181472
Martínez-Labarga, Cristina; Rickards, Olga. La utilización del DNA antiguo en la investigación de la historia evolutiva humana; Revista Española de Antropología Biológica. Sociedad Española de Antropología Biológica, Universidad de León, España: León, 1999; vol. 20, pp. 195-213.
Mattoso, José. Introdução: Legitimação e linhagem, e-Spania. 2011; Available online: http://journals.openedition.org/e-spania/20258 (accessed on 17 May 2019).
Mutation Surveyor® software. n.d. (Copyright 2011–2012). Available online: http://www.softgenetics.com/mutationSurveyor.html (accessed on 9 January 2023).
Olalde, Íñigo; Mallick, Swapan; Patterson, Nick; Rohland, Nadin; Villalba-Mouco, Vanessa; Silva, Marina; Dulias, Katharina; Edwards, Ceiridwen; Gandini, Francesca; Pala, Maria et al. The genomic history of the Iberian Peninsula over the past 8000 years. Science; 2019; 363, pp. 1230-34. [DOI: https://dx.doi.org/10.1126/science.aav4040]
Olasz, Judit; Seidenberg, Verena; Hummel, Susanne; Szentirmay, Zoltán; Szabados, György; Melegh, Béla; Kásler, Miklós. DNA profiling of Hungarian King Béla III and other skeletal remains originating from the Royal Basilica of Székesfehérvár. Archaeological and Anthropological Sciences; 2019; 11, pp. 1345-57. [DOI: https://dx.doi.org/10.1007/s12520-018-0609-7]
Pääbo, Svante; Poinar, Hendrik; Serre, David; Jaenicke-Despres, Viviane; Hebler, Juliane; Rohland, Nadine; Kuch, Melanie; Krause, Johannes; Vigilant, Linda; Hofreiter, Michael. Genetic analyses from ancient DNA. Annual Review of Genetics; 2004; 38, pp. 645-79. [DOI: https://dx.doi.org/10.1146/annurev.genet.37.110801.143214] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/15568989]
Parson, Walther; Dür, Arne. EMPOP-A Forensic mtDNA database. Forensic Sci International: Genetics; 2007; 1, pp. 88-92. [DOI: https://dx.doi.org/10.1016/j.fsigen.2007.01.018]
Pastor, Juan Francisco; De Paz, Felix; Barbosa, Mercedes; García, Natividad; Sanjosé, Isabel; Palomo-Díez, Sara; Arroyo-Pardo, Eduardo. Natural mummifications in Castile & Leon (Spain). European Journal of Anatomy; 2021; 25, pp. 221-32.
Phillips, Chris; Salas, Antonio; Sánchez, Juan; Fondevila, Manuel; Gómez-Tato, Antonio; AntonioAlvarez-Dios, José; Calaza, Manuel; de Cal, María Casares; Ballard, David; Lareu, María Victoria et al. Inferring ancestral origin using a single multiplex assay of ancestry-informative marker SNPs. Forensic Science International: Genetics; 2007; 1, pp. 273-80. [DOI: https://dx.doi.org/10.1016/j.fsigen.2007.06.008] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/19083773]
Rollo, Franco; Ermini, Luca; Luciani, Stefania; Marota, Isolina; Olivieri, Cristina; Luiselli, Donata. Fine characterization of the Iceman’s mtDNA haplogroup. American Journal of Physical Anthropology; 2006; 130, pp. 557-64. [DOI: https://dx.doi.org/10.1002/ajpa.20384]
Ruiz, Yarimar; Phillips, Chris; Gomez-Tato, Antonio; Alvarez-Dios, José Antonio; de Cal, Manuel Casares; Cruz, R.; Maroñas, Olalla; Söchtig, Jens; Fondevila, Manuel; Rodriguez-Cid, M. J. et al. Further development of forensic eye color predictive tests. Forensic Science International: Genetics; 2013; 7, pp. 28-40. [DOI: https://dx.doi.org/10.1016/j.fsigen.2012.05.009] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/22709892]
Salerno, Alfredo; Lago, Giampietro; Berti, Andrea; Calì, Francesco; Mirisola, Mario; Carta, Patrizia; Ferro, Carmen; Romano, Valentino. mtDNA analysis of the human remains buried in the sarcophagus of Federico II. Journal of Cultural Heritage; 2005; 6, pp. 313-19. [DOI: https://dx.doi.org/10.1016/j.culher.2005.07.001]
Stolarek, Ireneusz; Juras, Anna; Handschuh, Luiza; Marcinkowska-Swojak, M.; Philips, A.; Zenczak, M.; Debski, A.; Kócka-Drenz, H.; Piontek, Janusz; Kzlowski, Piotr et al. A mosaic genetic structure of the human population living in the South Baltic region during the Iron Age. Scientific Reports; 2018; 8, 2455. [DOI: https://dx.doi.org/10.1038/s41598-018-20705-6]
Vai, Stefania; Brunelli, Andrea; Modi, Alessandra; Tassi, Francesca; Vergata, Chiara; Pilli, Elena; Lari, Martina; Susca, Roberta Rosa; Giostra, Caterina; Baricco, Luisella Pejrani et al. A genetic perspective on Longobard-Era migrations. European Journal of Human Genetics; 2019; 27, pp. 647-56. [DOI: https://dx.doi.org/10.1038/s41431-018-0319-8]
van Oven, Mannis; Kayser, Manfred. Updated Comprehensive Phylogenetic Tree of Global Human Mitochondrial DNA Variation. Human Mutation; 2009; 30, pp. E386-E394. [DOI: https://dx.doi.org/10.1002/humu.20921]
van Oven, Mannis. PhyloTree Build 17: Growing the human mitochondrial DNA tree. Forensic Science International: Genetics Supplement Series; 2015; 5, pp. 9-11. Available online: http://www.phylotree.org (accessed on 9 January 2023). [DOI: https://dx.doi.org/10.1016/j.fsigss.2015.09.155]
Vargas Blanco, Rufino. En torno a la boda del Infante don Felipe de Castilla y doña Cristina, Infanta de Noruega. Boletín de la Institución Fernán González; 1968; 171, pp. 380-81.
Veeramah, Krishna R.; Rott, Andreas; Groß, Melanie; Dorp, Lucy van; López, Saioa; Kirsanow, Karola; Sell, Christian; Blöcher, Jens; Wegmann, Daniel; Link, Vivian et al. Population genomic analysis of elongated skulls reveals extensive female-biased immigration in Early Medieval Bavaria. Proceedings of the National Academy of Sciences; 2018; 115, pp. 3494-99. [DOI: https://dx.doi.org/10.1073/pnas.1719880115] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/29531040]
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
Details
; Gomes, Cláudia 1
; Fondevila, María Sonia 2 ; Esparza-Arroyo, Ángel 3 ; López-Parra, Ana María 1
; Lareu, María Victoria 2 ; Arroyo-Pardo, Eduardo 1
; Pastor, Juan Francisco 4
1 Health Legislation, Psychiatry, and Pathology Department, Medicine Faculty, The Complutense University of Madrid, 28040 Madrid, Spain; Forensic Sciences Group, Genetics and Toxicology, San Carlos Research Institute (IdISSC), Clínico San Carlos Hospital, 28040 Madrid, Spain
2 Medicine and Odontology Faculty, Santiago de Compostela University, 15705 Santiago de Compostela, Spain
3 GIR ‘PrehUSAL’, Department of Prehistory, Ancient History and Archaeology, University of Salamanca, 37007 Salamanca, Spain
4 Anatomy and Embriology Department, Medicine Faculty, Valladolid University, 47003 Valladolid, Spain