The genetics and genealogy research led by the University of Leicester discovered that the male line of descent is broken at one or more points in the line between Richard III and Henry Somerset, 5th Duke of Beaufort (1744-1803).
This provides scientific evidence for the first time of a possible question mark over medieval lines of succession in the monarchy, the researchers said.
Critically, there is no evidence whatsoever on when the break occurred, however, it is statistically far more likely that the break in the male line occurred lower down the chain - and therefore not affecting any Royal lineage – including that of the present day.
Historically Royal succession has taken a number of twists and turns over the centuries and is not based on straight linear inheritance, researchers said.
However, the research does pose a question mark over where the break might have occurred - and the potential implications for the historical monarchy.
"There are one or more breaks in the chain from Richard III to Henry 5th Duke of Beaufort," Professor Kevin Schurer said.
There are 19 links in that chain, so there is an equal probability of any one of those 19 being broken. From a historical view point, where the break occurred has differential consequences.
"There are five links in that chain between Richard, up through Edward, and then to John of Gaunt. And then the remaining majority of the links are down through the Somersets and the Beaufort line," said Schurer.
"Now if - and it is a very big IF - the break in the chain is one of those five, between John of Gaunt and Richard III, historians could ask questions - theoretically – about the inheritance of a number of the Plantagenet monarchs.
And since Henry Tudor's mother was also a Beaufort descended from John of Gaunt, there is a question here too. "However, statistically speaking, the break is far more likely to have occurred in the larger part of the chain which does not affect any of the different Royal lines of succession
at all," Schurer said.
Schurer, who is the Pro-Vice-Chancellor for Research at the University of Leicester, said "there was absolutely no evidence that a link in the Royal chain of succession had actually been broken."
"The fact that we do not find a match between the living male-line relatives and the skeletal remains is not at all surprising to me," Dr Turi King said.
"We knew from work that I, and others, have carried out in the past that the incidence of false-paternity, where the biological father is not the supposed father, is historically in the region of 1-2 percent per generation," said King. The finding was published in the journal Nature Communications.