Genetic investigation of the fetus found in grave 4, and its family relation to person II/52_3, identified as

Béla II (the Blind)

1. Archaeological data. In the immediate vicinity of the grave of Béla III and Anne of Antioch, at the feet of the royal couple, Érdy also found a fourth and a fifth grave (Figure 13). According to contemporary customs, only family members were buried next to royalty. In the fourth earth grave, in a wooden coffin sunk into the ground laid the skeletons of a woman and a fetus. The earth grave was around the same age as Béla III’s grave. The female skeleton buried in the coffin was lost, but we still have the fetus found among the pelvic bones, it is registered under number II/52_4 (by Éry and her working group). According to archaeologists’ opinions, it may have been Béla III’s grandchild in grave 4, but the current genetic investigation suggests that it was Princess Predslava of Kiev. The fragmented bones excavated next to the earth grave from a wooden coffin in a brick-lined grave were lost. In view of the contemporary burial customs, we believe that the person in the 5^th grave may have been Predslava’s husband, Prince Álmos of the Árpád Dynasty.

2. Genetic investigation of the fetus’ skeleton. Up to this point, the identities of the persons in the fourth and fifth grave were unknown, because only the skeleton of the fetus from the 4^th grave was not lost, and thus genetic investigations had to be carried out on it alone. The fetus was identified as male in the Göttingen laboratory. We confirmed twice that that the gender determination was incorrect and that in fact the fetus was female (Olasz, J and Csernák, E). A-STR studies were conducted on the fetus’ bones using PCR methods in Göttingen and

Budapest. In the Göttingen laboratory, the A1 and A2 alleles could only be determined in the case of a total of 3 markers (D5S818, FGA, TH01), each of these markers’ alleles matched one of skeleton II/52_3’s (maternal) alleles. In the other three cases (D10S, D18S51, D21S51), only one of the alleles of the A-STR markers could be determined, none of which were identical to skeleton II/52_3’s A-STR markers.

The A-STR marker investigation conducted in the Budapest-2 laboratory by Dr Erzsébet Csernák covered 15 markers, out of which in 11 cases we were able to detect both alleles, in two cases we were able to detect one allele, and in another 2 cases we did not arrive at any evaluable results. We evaluated the individual results obtained from the A-STR markers via PCR with next generation sequencing, and in the comparisons, we took into account the sequencing (NGS) data. After summarizing the results of the analyses conducted by the Göttingen and Budapest laboratories and supplementing them with the CSF1PO marker data examined by Dr Judit Olasz, it immediately becomes clear, that out of the 16 A-STR markers which can definitely be evaluated, only markers D10S1248, D21S11 and D22S1045 failed to match with either alleles of the II/52_3 consensus A-STR marker.

Extremely long A-STR alleles can be found at the non-matching D21S11 marker, similarly to the SE33 marker which was excluded due to PCR artifacts (see Table 16). The possibility of a PCR artifact arises with a high probability for marker D21S11 as well, and thus going forward we do not take these data into account either. Thus, ultimately we only evaluated the results of fifteen A-STR markers. In a ratio of 13/15, the aggregate A-STR marker results were identical to one of skeleton II/52’s alleles (the maternal one): this points to a close family relationship between the fetus and skeleton II/52 (Table 20).

Árpád Dynasty consensus II/52 A-STR

Fetus,

Göttingen Fetus, Budapest II/52 mtDNS haplogroup D2S441 10 10 n.a. n.a. 11.3 10** sequencing D2S1338 20 18 n.a. n.a. 17** 20** sequencing

D3S1358 n.a. n.a. 15* 16* 15* 16*

D3S1358 14 14 n.a. n.a. 14** 14** sequencing D5S818 10 12 12 13 n.a. n.a. n.a. n.a.

D7S820 8 9 n.a. n.a. 8** 9** sequencing D8S1179 12 14 n.a. n.a. n.a. 13* n.a. n.a.

D19S433 13 13 30.2 n.a. 13** 13** sequencing D21S11 30 32.2 30.2 - 29.2* 30.2* n.a. 30.2*

Table 20 Comparing the A-STR markers of the fetus with skeleton II/52_3’s similar markers. Matching alleles of the A-STR investigation conducted on the fetus’ skeleton in Göttingen and Budapest with the A-STR alleles of skeleton II/52_3 (highlighted in bold numbers and yellow). *: PCR-determined data; **: Sequencing data; ***: The A-STR marker CSF1PO was only investigated by Dr Olasz in the Budapest-1 laboratory, her data are displayed in the table; n.a.: no data; n.v.a.: no valid data; A1/2: allele.

The mtDNA HVR 1/2 genetic investigation of the fetus’ skeleton was conducted by Dr Olasz. She found that the fetus could not be Béla III’s grandchild (one of his daughter’s children), as its mitochondrial DNA differs from that of Anne of Antioch. The fetus does not have the 16240G specific to the Queen, and the Queen does not have 16294T and 16296T SNV found in the fetus.

According to Balázs Holczmann’s hypothesis, if the maternal alleles of the fetus match with the maternal alleles of skeleton II/52_3 (they match in a ratio of 13/15), or its mtDNA haplogroup is T2b2b1, then the female skeleton is Predslava of Kiev, while skeleton II/52_3 is Béla II (the Blind) (Holczmann 2019a). The sequence variations 16194T and 16296T found in the mtDNA HVR-1 region of skeleton II/52 determine the fetus’ T2b haplogroup by themselves (Pike et al.

2010). Thus, the fetus’ maternal genetic markers and mitochondrial haplogroup match the maternal markers of skeleton II/52. This is possible only if the mother of the fetus is Predslava of Kiev, and the fetus is Béla II’s sibling (see Figure 44). 16294T SNV can be found not only in the fetus, but in skeleton II/52 (Béla II) as well, which serves as further proof that the fetus is in fact Béla II’s sibling.

The Ukrainians are proud of their royal connections and published a series of stamps on the Kievan wives of European Kings. Among these, the wives of Hungarian Kings were the following: Euphemia of Kiev, wife of Kálmán; Anastasia of Kiev, wife of András I; Predslava of Kiev, wife of Álmos; and Euphrosyne of Kiev, wife of Géza II (Figure 43). Predslava married Prince Álmos of the Árpád Dynasty, their later fate is still unknown.