The identity of King Richard III’s remains confirmed using Ion Torrent technology

A king’s remains discovered in a car park

King Richard III had a short but notable reign (1483-1485), starting with the death of his brother, his 12-year old nephew Edward V declared an illegitimate heir to the throne, then Richard III ascending to the throne. His tumultuous reign ended when he was killed in battle by the forces of Henry VII.  At the age of 32, he was the last of English Royalty to die in battle, and the only king whose remains were not known.

His reputation as a villain was secured by Shakespeare, who wrote the play Richard III about a hundred years after his death.

A public-private partnership

A historical group called the Richard III Society was organized around furthering research ‘dedicated to reclaiming the reputation of a king of England who died over 500 years ago’. Based upon evidence uncovered by historians to suggest that he was buried on the site of the Grey Friers Friarage in Leicester, they raised funds for an archaeological dig. Working with the Leicester University, a unique private / public partnership began based upon the evidence that the friary was originally located underneath the Leicester City Council parking lot.

In September of 2011 three long trenches were dug. Early on the Grey Friarage Chapel boundaries were identified, and a skeleton unearthed labeled ‘Skeleton 1’. Subsequent analysis revealed via radiocarbon dating and physical forensic evidence (such as damage to the skull and scoliosis) led the archeologists in February of 2013 to conclude that this skeleton was indeed that of King Richard III.

A positive mitochondrial DNA sequence identification

Now in December of 2014 a genetic analysis of ‘Skeleton 1’ positively identifies it as the remains of Richard III, using the Ion Torrent PGM™ System to sequence the remain’s mitochondrial DNA (mtDNA). It is fortunate that two direct descendants from Richard III’s sister Anne of York have been identified, some seventeen or nineteen generations later, to make the maternal lineage link. (It is noteworthy that a detailed geneology of both ML1 and ML2 are included in the publication’s Supplement.) Of the two samples ML1 and ML2, one was a perfect match for the entire mtDNA genome, and the other was a single mismatch (the human mtDNA genome is about 16.6kb in length).

In addition, autosomal SNPs determining hair and eye color were also identified from this sample using Ion Torrent technology, using what is called HIrisPlex SNPs (HIrisPlex are eye- and hair-color determining SNPs described by Manfred Kayser and colleagues, who recently spoke at the Ion World Manchester event.). King Richard III’s eyes were determined to be blue, which agrees with the best scholarship of surviving portraiture. Many portraits of Richard III were either modified from existing portraits in the following years after his death (to make him more sinister-looking, either from the angle of his face or by making his fingers look claw-like), and later portraits were derivatives of these. His hair genetically was determined to be blond, which also differs from the portraiture; however there are instances where blond hair at birth changes in color during development.

One intriguing wrinkle arose in the analysis of the Y-chromosome; there was no match to five 19^th-generation descendants on the patrilineal side; the authors note that 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’. For those curious about how often this occurs, an estimated average false-paternity rate has been determined by other researchers to be on the order of ~1-2%.

This work was published recently in Nature Communications, titled “Identification of the remains of King Richard III”.

For those in Europe who are interested in forensics, Thermo Fisher Scientific will be sponsoring a two-day conference called “Human Identification Solutions Conference” in Madrid, Spain March 5-6 2014.

Reference: King TE, Fortes GG, Balaresque P, Thomas MG, Balding D, Delser PM, Neumann R,  Parson W, Knapp M, Walsh S, Tonasso L, Holt J, Kayser M, Appleby J, Forster P, Ekserdjian D, Hofreiter M, Schürer K. Identification of the remains of King Richard III. Nat Commun. 2014 5:5631

 

doi: 10.1038/ncomms6631