As a follow up of the mtDNA Haplogroup H tutorial, lets take a look at a fun topic and see how anthropologists have used mtDNA to help solve historical questions. Let's examine some historical figures who may have belonged to Haplogroup H, one of the largest European Haplogroups.
Who was she?
Empress Alexandra Feodorovna of Russia, the granddaughter of Queen Victoria was born on June 6, 1872 as Princess Alix of Hess and by Rhine in Darmstadt (Germany). She was the sixth child of Grand Duke Louis IV of Hesse and by Rhine and Princess Alice of the United Kingdom. She was the princess consort of Nicholas II, and the last Tsaritsa of Russia.
What were the anthropologists trying to find out? ie why was her DNA tested?
At the end of the February Revolution of 1917, Tsar Nicholas II was forced to abdicate. The Romanov family were imprisoned in Ipatiev House at Ekaterinburg in the Urals of Central Russia in 1918 with three servants and the family doctor. On the night of July 16, 1918, the entire Romanov family was shot and killed by the Bolshevik firing squad.
According to legend, two bodies were burnt and the others were buried in a roadside pit. To hinder identification, sulphuric acid was said to have been thrown into the open grave and a truck was driven back and forth over the grave.
The case remained a mystery until July 1991, when nine human skeletons were discovered in a shallow pit around 20 miles from Ekaterinburg, Russia. Historians speculated that the remains belonged to the Romanov family. To help solve the mystery, DNA testing was performed to identify the remains.
Who’s who in researching the history of the Romanov family:
Top peer reviewed research publications for Tsarina Alexandra and the Romanov family:
This table lists the most significant papers for the Romanov family and the DNA tests that were performed to identify their remains in peer reviewed journals. Links are provided to access the original articles.
These papers provide the extent of what is known today about the DNA type of Tsarina Alexandra and provides answers to the question of whether the bodies discovered in the grave belong to the Romanov Family.
|Name of Scientific Article||Scientific Journal|
|Identification of the remains of the Romanov family by DNA analysis. Gill et al, Forensic Science Service, Berkshire, UK. Click here to read abstract||Nat Genet. 1994 Feb; 6(2):130-5.|
|DNA forensics. Buried, recovered, lost again? The Romanovs may never rest.Stone et al. This publication is published in Science magazine, a subscriptions only magazine. Click here to go to Science magazine.||Science. 2004 Feb 6; 303(5659):753.|
|Molecular, forensic and haplotypic inconsistencies regarding the identity of the Ekaterinburg remains. Knight et al, Stanford University, Stanford, USA. Click here to read the article||Ann Hum Biol. 2004 Mar-Apr; 31(2):129-38.|
|Ongoing controversy over Romanov remains.Hofreiter et al, Gill et al, Knight et al. This publication is published in Science magazine, a subscriptions only magazine. Click here to go to Science magazine.||Science. 2004 Oct 15;306(5695):407-8, 408-9, 409-10|
As more articles become available, they will be listed here.
Solving the case: Collecting DNA samples from the Ekaterinburg remains
The first step in solving this case is to test the DNA of the Ekateringburg remains. After the DNA type of the remains is known, it can be compared to the DNA type of known relatives in the royal family. Since Tsarina Alexandra is the maternal granddaughter of Queen Victoria (Queen Victoria is her mother’s mother), Tsarina Alexandra must have inherited her mtDNA from Queen Victoria. A living descendent of Queen Victoria who carries Queen Victoria’s mtDNA type is Prince Philip. The following diagram shows in purple how the mtDNA is passed down from Queen Victoria to her descendents and which one of her descendents carry her mtDNA:
Individuals who inherited the mtDNA type of Queen Victoria are highlighted in purple.
The descendent tree for Queen Victoria indicates that Tsarina Alexandra should have the same mtDNA profile as Prince Philip as well as other living and deceased descendents from the same line. By collecting the DNA sample from known family members, scientists can find out the expected mtDNA type for Tsarina Alexandra and her children and use it to identify the remains.
Who was tested to solve this case?
Three Types of DNA tests were performed to confirm the identity of the Ekateringburg remains:
Step 1: Autosomal STR Testing: To determine whether the 9 skeletons are related to each other
Autosomal STR testing (the same technology used for paternity testing) was used to determine whether the nine skeletons are from the same family.
Autosomal DNA Markers
|3||15, 16||8, 10||5, 7||12, 13||11, 32||Child|
|4||15, 16||7, 10||7, 7||12, 12||11, 32||Parent|
|5||15, 16||7, 8||5, 7||12, 13||11, 36||Child|
|6||15, 16||8, 10||3, 7||12, 13||32, 36||Child|
|7||15, 16||8, 8||3, 5||12, 13||32, 36||Parent|
The results of the autosomal test confirmed the following:
This information is consistent with the skeletons belonging to members of the Romanov family.
Step 2: Determining the gender of the skeletons
The next set of tests sets out to determine the gender of the skeletons.
|Skeleton #||Sample Type||Sample Quality||Gender||Relationship|
The results of the gender determination tests showed that four of the skeletons are male and one was female. The combined results of the autosomal test and gender test leads to the speculation that skeleton #4 is the father, skeleton #7 is the mother, and skeletons 3, 5, and 6 are their daughters.
Step 3: What if the skeletons belonged to the Romanov family?
If the remains truly belong to the Romanov family, then:
|Skeleton #||Relationship||Speculated Identity (if the Romanovs)|
|4||Father||Tsar Nicholas II|
The next step is to confirm the speculated identities by comparing each skeleton to living royal family members.
Step 4: Solving the mystery. Confirming the identity of Skeleton #7, Tsarina Alexandra and her daughters, Skeletons 3, 5, and 6
Now that the preliminary studies indicate the possible identities of each of the skeletons, the next step is to confirm the identity by comparing to the DNA of living royal family members. Tsarina Alexandra is the grandaughter of Queen Victoria, and her mtDNA should be identical to the mtDNA of Prince Philip, a living descendent of Queen Victoria.
The next step is to test the mtDNA of Prince Philip and compare it to the mtDNA of the skeletons.
Prince Philip’s mtDNA:
|Name||HVR1 Region||HVR2 Region||Coding Region||Publication|
|Prince Philip||16111,16357||263,315.1||Not tested||Gill et al 1994|
Prince Philip is a direct descendent of Queen Victoria and he carries the mtDNA type of Queen Victoria.
The mtDNA of the Skeletons:
|Name||HVR1 Region||HVR2 Region||Coding Region||Publication|
|Skeleton #3:Putative daughter||16111,16357||263,315.1||Not tested||Gill et al 1994|
|Skeleton #4:Putative Tsar Nicholas II||16126,16169Y,16294,16296||73,263,315.1||Not tested||Gill et al 1994|
|Skeleton #5:Putative daughter||16111,16357||263,315.1||Not tested||Gill et al 1994|
|Skeleton #6:Putative daughter||16111,16357||263,315.1||Not tested||Gill et al 1994|
|Skeleton #7:Putative Tsarina Alexandra||16111,16357||263,315.1||Not tested||Gill et al 1994|
Conclusion: The results of the mtDNA test confirm the following:
In summary, the DNA studies confirm that five out of the nine skeletons discovered truly belonged to the Romanov family, namely Tsar Nicholas II, Tsarina Alexandra, and 3 of their 4 daughters. The skeleton of one of their daughters and their only son, Alexei, remain missing.
The Aftermath: Doubts and Controversies
Since the findings, the skeletons were reburied with honors in the imperial-era capital of St. Peterburg. The two remaining children remain missing.
The scientific findings and results have since been challenged by Knight et al, suggesting irregularities and sample handling deficiencies.
Recent Findings from the summer of 2007
In the summer of 2007, using metal detectors and metal rods as probes, two burned partial skeletons were discovered at a bonfire site near Ekaterinburg, 900 miles east of Moscow. 15 intact bone fragments and more than 40 pieces of charred bone were discovered.
The site appeared to match the site described in Yurovsky’s (the Bolshevfik officer in charge of the Romanovs’ captivity) memoirs. According to his memoirs, the bodies of nine victims were doused with sulphuric acid and buried along a road. Alexei’s body and one of his sister’s bodies was burned and left in a pit nearby
Preliminary analysis of the bones showed that the remains were from a boy roughly between the ages of 10 to 13 years old at the time of death, the putative Alexei and a young woman roughly between the ages of 18 to 23 years old at the time of death, attributed to one of the Romanov daughters, likely Maria or Anastasia.
Genetic investigations were performed in the Sverdlovsk Regional Forensic Medicine Bureau, a lab in Moscow, and a US laboratory, the University of Massachusettes Medical School.
On April 30, 2008, the groups claimed that DNA testing (mainly mitochondrial DNA analysis) proved that the remains belonged to Tsarevich Alexei and one of his sisters (likely Maria) were released in several major newspapers and news sources (including New York Times, BBC, and MSNBC).
|Bones turn up in hunt for last czar’s son||2007|
|Probe reopened into death of last Russian czar||2007|
|DNA tests may solve mystery of last czar’s heirs||2008|
|Report: DNA proves link to last czar’s kids||2008|
|Russia dig finds ‘tsar’s family’||2007|
|Lost Romanov bones ‘identified’||2007|
|Tsar’s lost children identified||2008|
|Experts May Have Found Remains of Czar’s Children||2008|
|Amateurs Unravel Russia’s Last Royal Mystery||2007|
|DNA Tests Confirm the Deaths of the Last Missing Romanovs||2007|
What is the haplogroup of Queen Victoria’s descendents?
Now that the mtDNA type of Queen Victoria’s line is known, amateur genealogists from around the world have used the data to compare to their own families to see if they may have links to royalty.
Although the original researchers never attempted to use the data from the study to determine the haplogroup of this royal family line, amateur genealogists have tried t use the data from the study to determine the mtDNA haplogroup of Queen Victoria’s line, concluding that Queen Victoria and her descendent Tsarina Alexandra belonged to Haplogroup H. Let’s take a look at the raw data and see how accurate they are.
Step 1: Click here to download and print the mtDNA Haplogroup map so that you can follow along with the discussion.
Step 2: Identify the presence and absence of HVR1 markers on the map.
Tsarina Alexandra only had two HVR1 markers, namely 16111 and 16357. On the map, all HVR1 markers are in blue. Starting from the CRS, move outwards and cross off all of the HVR1 markers that Tsarina Alexandra does not have, circle the ones that she does have and put a question mark next to the ones that are unknown:
The results of the HVR1 test helps to eliminate the haplogroups that Tsarina Alexandra definitely does not belong to, and shows that Tsarina Alexandra must belong to either Haplogroup R, Pre-HV, HV, H, or the CRS branch of H.
Step 3: Identify the presence or absence of HVR2 markers on the map:
Tsarina Alexandra has 2 markers in her HVR2 region, namely 263 and 315.1. On the map, all HVR2 markers are red. Starting from the CRS, move outwards and cross off all of the HVR2 markers that Tsarina Alexandra does not have and circle all of the HVR2 markers that she does have:
The results of the HVR2 test helps to eliminate Haplogroup R and shows that Tsarina Alexandra must belong to either Haplogroup Pre-HV, HV, or H. Tsarina Alexandra has marker 263 which brings her away from CRS, indicating that she is unlikely to belong to CRS, but the presence of 263 cannot eliminate her from CRS as such a conclusion woudl require more data from the coding region.
In conclusion, the results of the scientific studies show that Tsarina Alexandra can belong to any one of the following Haplogroups:
These results are similar to those of Marie Antoinette, even though the exact mtDNA mutations that they carry are slightly different. To confirm whether Tsarina Alexandra is really a member of Haplogroup H, one would need to test the coding region of her mtDNA to confirm that she does not have markers 7028 and 14766. However, no coding region data was ever available for Tsarina Alexandra or for any of her living or deceased relatives, so there is no way to confirm that she is a member of Haplogroup H. Since the original researchers were aiming to identify her remains rather than to determine her haplogroup, the coding region was never examined and in this particular case, coding region SNP test is required to confirm her haplogroup.
In conclusion, it is pre-mature to conclude that Tsarina Alexandra is a member of Haplogroup H, but the data from the studies to date do serve to narrow down her possible haplogroups to H, HV, and Pre-HV. Further studies woud need to focus on markers 7028 and 14766 in the coding region which would provide a determination of which Haplogroup Tsarina Alexandra actually belongs to, and if she does in fact belong to Haplogroup H, further H subclade testing of markers in the Coding Region should confirm which branch of H she falls into.