Scientists are now 99.999 per cent sure that the skeleton with a twisted spine found in a Leicester car park in 2012 is that of the last Plantagenet king.
But while DNA analysis has confirmed two female relatives of the king living today, new findings show evidence of at least one break in the male line, which threatens to shake the foundations of the Tudor dynasty.
The skeleton’s male Y chromosome, which is passed only from father to son, did not match that of five living individuals who claim a paternal link with King Richard via their shared ancestor, Henry Somerset, the 5th Duke of Beaufort.
While a relatively recent break would have no royal significance, an illegitimacy dating back several centuries to the first Duke of Lancaster, John of Gaunt, or his son John Beaufort, Earl of Somerset, would cast doubt on the succession of a whole series of monarchs - including the Queen, whose ancestry can be traced back to the founder of the Tudor dynasty, King Henry VII, via James I and Mary, Queen of Scots.
But Professor Kevin Schurer, pro-vice chancellor of the University of Leicester, who co-led the research team, insisted: “We are not in any way indicating that Her Majesty should not be on the throne.”
John of Gaunt had two sons, John Beaufort and the first Lancastrian king Henry IV, whose direct descendants were Henry V and Henry VI. The Tudor line stems from John Beaufort who was Henry VII’s great grandfather.
Richard III was connected to these lineages through his great grandfather Edmund, Duke of York - John of Gaunt’s brother.
Prof Schurer said: “We don’t know where the break is, but if there’s one particular link that has more significance than any other, it has to be the link between Edward III and his son John of Gaunt.
“John of Gaunt was the father of Henry IV, so if John of Gaunt was not actually the child of Edward III, arguably Henry IV had no legitimate right to the throne, and therefore neither did Henry V, Henry VI, and, indirectly, the Tudors.”
Writing in the journal Nature Communications, the scientists said the claim to the crown of the “entire Tudor dynasty” partly rested on its members’ descent from John of Gaunt.
However, Prof Schurer stressed that the history of the British monarchy took “all kinds of twists and turns” and the Y chromosome discovery had no bearing on the present Queen’s right to rule. The Tudors took the crown essentially “by force” while using the blood line leading to John of Gaunt to back up their claim, he said.
Richard III was killed at the Battle of Bosworth in 1485, the last significant clash between the forces of the Houses of Lancaster and York in the War of the Roses.
In their paper, the researchers compared the investigation to a missing person case that becomes more difficult over time - in this case, 527 years.
According to historical records he was buried in Grey Friars Church, Leicester, which once stood on the site of the car park where his bones were found. Descendents of the King lost their passionate campaign to have the skeleton reinterred in York, rather than Leicester, in May. He will be buried at Leicester Cathedral in March.
Scientists from the University of York played a crucial role in helping to crack “the UK’s oldest forensic case”.
Dr Turi King, the University of Leicester’s lead researcher, carried out part of her work at York. Genetic analysis showed a 96 per cent probability that Richard had blue eyes and a 77 per cent likelihood that he was blond, at least in childhood.
Ancient DNA expert, Professor Michi Hofreiter, an Honorary Professor of Biology at York, who worked on the analysis, said: “It’s amazing how much we can deduce from ancient DNA today.
“Making inferences about hair or eye colour of a person just from some DNA snippets obtained from a skeleton would have been unthinkable just a few years ago.”