King Richard III, considered one of England's most notorious rulers, still delivers shocks to his country 529 years after his death, his twisted skeleton threatening to shake the foundations of the royal dynasty.
Genetic analysis of the bones found two years ago under a parking lot in Leicester have proven with 99.999 percent certainty that the remains do belong to the last Plantagenet King. However, some other royally embarrassing questions were raised in the process.
While mitochondrial genomes, inherited through the maternal line, showed a genetic match between the skeleton and two modern female-line relatives, it wasn't possible to trace a living relative on the male line through the Y chromosome, which is passed on from father to son.
It's Really Richard: DNA Confirms King's Remains
"The Y-chromosome haplotypes from male-line relatives and the remains do not match, which could be attributed to a false-paternity event occurring in any of the intervening generations," Turi King, a geneticist at the University of Leicester, and colleagues wrote in the journal Nature Communications.
In other words, somewhere along the more than 500-year-old male line, a king may have been cuckolded, his wife giving birth to another man's child.
The century old royal sex scandal could potentially undermine not only Richard III's claim to the throne, but also the British royal line of succession up to the current Queen Elizabeth II.
The analysis shows the non paternity, or break, occurred somewhere along the male line linking Richard III and Henry Somerset, the 5th Duke of Beaufort (1744-1803), who is the common ancestor of the living male-line relatives.
This Is What Killed Richard III
As Richard III died without any male heirs, King and colleagues had to trace the paternal line backwards to Edward III (1312-1377), Richard III's great great grandfather, and then move forward through the ancestral line of the Houses of Lancaster and Tudor. Overall, there are 19 generations between Richard III and the male-line related individuals who are alive today.
At least one of the 19 men in the chain was illegitimate.
"We don't know where in the chain the break occurs and therefore are not able to say whether it would have affected historical royal succession," King told Discovery News.
While a relatively recent break would question only the non-royal family of the current dukes of Beaufort, an illegitimacy near the top of the tree would raise question marks over a whole series of monarchs, including the reigning Queen Elizabeth II, whose ancestry can be traced back to King Henry VII, the founder of the Tudor dynasty, via James I and Mary, Queen of Scots.
King Richard III Feasted on Wine and Swans
"If there is 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," Kevin Schürer, professor of English Local History and Pro-Vice-Chancellor for Research at the University of Leicester, said.
Indeed, John of Gaunt was rumored to be illegitimate, with gossips suggesting he was the son of a Flemish butcher.
"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," Schürer said.
He stressed, however, that the study is not in any way questioning the current Queen's right to throne.
"The British monarchy took all kinds of twists and turns," he said.
The only way to solve the puzzle would be to exhume skeletons along the 19-link chain and check their DNA.
Richard III May Have Had a Brummie Accent
Richard III's remains are to be reburied in Leicester in March 2015.
Depicted by William Shakespeare as a bloodthirsty usurper, the medieval king ruled England from 1483 to 1485. He was killed in 1485 in the Battle of Bosworth, which was the last act of the decades-long fight over the throne known as War of the Roses.
England's last king to die in battle, Richard III was defeated by Henry Tudor, who became King Henry VII.
Image: King Richard III's skeleton as it was found in the car park two years ago. Credit: University of Leicester.