DNA tests of bones found in parking lot reveal new details about King Richard III

Researchers know he died in battle and had back trouble. Now, DNA analysis has shed light on what Richard III looked like -- and raised surprising questions about the rightful passage of the crown after his tragically short reign ended in 1485.

In a study published Tuesday in Nature Communications, researchers said the DNA analyses found that the last English king to die in battle, some 500 years ago, most likely had blue eyes and blond hair, closing more of the historical gaps in what we know of the mysterious monarch.

A twisted skeleton was first discovered under a church parking lot back in 2012, setting off a CSI-like investigation into whether it was indeed Richard III. Five months later, DNA evidence proved it was the king.

The new research carved out previously unknown genetic details about the royal find.

"We were able to zoom in on 11 genes with 24 little differences in those genes that can help us predict what somebody's hair and eye color is going to be," said University of Leicester geneticist Turi King, who in earlier work had found matches between DNA from the skeleton and two direct descendants of Richard III on the female line.

"He has got a 96 percent probability that he had blue eyes - so a very high probability he had blue eyes - and a 77 percent probability of blond hair," she said. "This would have been childhood hair color. It can darken with age. But taking the genetic predictions, we would suggest the most closely matching portraits is the arch-framed portrait in the Society of Antiquaries of London [shown at left] because it has blue eyes and a lighter color of hair."

Other portraits, which depicted the king with dark eyes and dark hair, have been proved less accurate.

King Richard III was immortalized by Shakespeare as a villainous hunchback who had his brother and nephews killed to secure the throne for himself. The discovery of what was believed to be his long-lost grave two years ago, revealed a skeleton that did, in fact, suffer from scoliosis or curvature of the spine. In February 2013, DNA tests confirmed that the battle-scarred, curved bones were indeed those of Richard III.

And in September of this year, scientists released whole body CT scans and micro-CT imaging of injured bones that revealed how he died.

According to those findings the king sustained a total of 11 wounds at or near the time of his death at the Battle of Bosworth Field on August 22, 1485. Nine of the injuries were inflicted to the skull, which suggests that he had removed or lost his helmet. Two other wounds were found elsewhere on the body.

In the latest study, researchers produced the most complete DNA analyses of Richard III, which affirmed that there was a "perfect mitochondrial DNA match" between the sequence obtained from the skeleton and two living relatives, previously identified. Michael Ibsen and Wendy Duldig were linked to Richard III through a female-only line descended from the king's eldest sister Anne of York.

Prof. Kevin Schurer, of the University of Leicester Centre for English Local History, told CBS News that this shows beyond a shadow of a doubt that the theory is true: These are the bones of the king.

And yet it hasn't silenced detractors who remain unconvinced. Tracing the bones to relatives, they say, is not sufficient to support the statement.

"Does this solve the issue? No, because the tests prove the relationship of the bones to the modern relatives 17 generations away, not to Richard III alone," Michael Hicks, professor Emeritus of Medieval History University of Winchester, said in an e-mail interview. "I don't question the scientific quality of the DNA analysis, but the conclusions."

King responded that doubters "take all the strands of evidence isolation but don't put them together which is what we've done."

"It's like a missing persons case. You would never rely just on genetic evidence or anything," she said. "We do excavations and we find a youngish male age 30-34 in the [Church of the Grey Friars] with battle injuries, severe scoliosis of the spine and the radiocarbon dates match. Then, we get a mitochondrial DNA match and we put all those strands of evidence together and it's between 99.999 and 99.99999 percent chance that this was Richard."

(The king was last seen in the Church of the Grey Friars, died in battle at 32 and had one shoulder higher than the other.)

The researchers acknowledge one part of the puzzle that doesn't fit: The Y-chromosome of the remains didn't match those from male-line relatives. But they said that "wasn't remarkable" given that a false-paternity event "could have occurred in any of the intervening generations."

Though unremarkable in the context of a positive identification, the mismatch could have important implications about Richard III's line of succession.

Given that a Y-chromosome is passed from a man to his male descendants, and does not change over time, the researchers said this could "be of key historical significance" particularly if it "occurred in the five generations between John of Gaunt and Richard III."

"A false paternity between Edward III and John would mean that John's son, Henry IV and Henry's direct descendant would have had no legitimate claim to the crown," the researchers wrote. "This would hold true, indirectly, for the entire Tudor dynasty."

King said the next step is finishing the complete genome sequence of Richard III by next year. It would be the first time the genome of a historical figure was sequenced and could offer up more tantalizing details of his life.

"It depends on how much DNA we get back and what sort of coverage we get," King said. "We know there are genes involved in predisposition towards heart disease or diabetes and even section of the genome thought to be involved in predisposition towards Scoliosis. That would be an interesting thing to look at."