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The future James VI of Scotland was the only son of Mary I, Queen of Scots, and her second husband, Henry Stuart, Lord Darnley. Mary’s rule over Scotland was insecure, and she and her husband, being Roman Catholics, faced a rebellion by Protestant noblemen. During Mary’s and Darnley’s difficult marriage, Darnley secretly allied himself with the rebels and conspired in the murder of the Queen’s private secretary, David Rizzio, just three months before James’s birth.

James’s father, Darnley, was murdered on February 10, 1567 at Kirk o’ Field, Edinburgh, perhaps in revenge for the killing of Rizzio.

James inherited his father’s titles of Duke of Albany and Earl of Ross. Mary was already unpopular, and her marriage on May 15, 1567 to James Hepburn, 4th Earl of Bothwell, who was widely suspected of murdering Darnley, heightened widespread bad feeling towards her.

In June 1567, Protestant rebels arrested Mary and imprisoned her in Loch Leven Castle; she never saw her son again. She was forced to abdicate on July 24, 1567 in favour of the infant James and to appoint her illegitimate half-brother, James Stewart, Earl of Moray, as regent.

The infant King James VI was anointed King of Scotland at the age of thirteen months at the Church of the Holy Rude in Stirling, by Adam Bothwell, Bishop of Orkney, on July 29, 1567. The sermon at the coronation was preached by John Knox. In accordance with the religious beliefs of most of the Scottish ruling class, James was brought up as a member of the Protestant Church of Scotland.