In 1561, nineteen-year-old Mary Stuart, Catholic Queen of Scotland, returns to her home country from France following the death of her husband, Francis II, to take up her throne, where she is received by her half brother, the Earl of Moray. In neighbouring England, Mary's twenty-eight-year-old cousin Elizabeth is Protestant Queen of England – unmarried, childless, and threatened by Mary's potential claim to her throne. Mary dismisses the cleric John Knox from her court. A Protestant and leader of the Scottish Reformation, Knox views Mary as a danger to the kingdom's Protestant supremacy.
Seeking to weaken her cousin's threat to her sovereignty, Elizabeth arranges for Mary, whom English Catholics recognise as their rightful Queen, to be married to an Englishman. She chooses Robert Dudley, whom she secretly loves; although he and Mary are unwilling, news of Elizabeth's smallpox convinces Mary to accept, provided she is named Elizabeth's heir apparent. Reluctant to let go of Dudley, Elizabeth sends Lord Darnley to Scotland with the pretence of living under their religious freedom. Despite sensing the ulterior motive, Mary grows fond of Darnley and eventually accepts his proposal.
Mary's impending marriage causes a constitutional crisis within both realms: Elizabeth is advised to oppose the marriage for fear that Darnley, an English noble, will elevate Mary's claim to the Crown; Mary's council is suspicious of Darnley, fearing an English takeover. Both kingdoms demand his return to England but Mary refuses, leading Moray to mount a rebellion against her. Mary marries Darnley, only to discover him in bed with her friend and private secretary, David Rizzio. Mary quashes the rebel forces but spares Rizzio and Moray, and demands that Darnley give her a child. When a child is conceived, Mary declares the child is "heir to Scotland and England" – offending the English.
Moray colludes with Darnley's father Matthew Stewart, 4th Earl of Lennox, to undermine Mary. They spread rumours that her child was illegitimately fathered by Rizzio, driving John Knox to vehemently denounce Mary as an adulteress. Fearing these accusations and the possible discovery of his homosexuality, Darnley is coerced by the underminers to join them in murdering Rizzio, and reluctantly delivers the final blow.
Discovering the plot, Mary agrees to pardon the conspirators if presented with evidence that Darnley took part. She ultimately forgives Moray, and asks Elizabeth to be her child's godmother. They agree the child is heir presumptive, despite the English court's hostility. Mary banishes Darnley but refuses to divorce him, despite the appeals of her council, which approaches her adviser and protector, the Earl of Bothwell, to have him killed.
After Darnley's death, Mary is forced to flee without her child. Bothwell advises that her council has decided she must marry a Scotsman immediately—and that Scotsman should be Bothwell himself. Mary resists, suspecting he was involved in Darnley's murder, but after he threatens her and subsequently rapes her, she consents. This induces Knox to preach to the Scots that Mary is a "harlot" who had her husband killed, leading Moray and her court to demand her abdication. Despite her objections, Mary eventually abdicates and flees to England.
Elizabeth arranges a clandestine meeting, where Mary asks for help to take back her throne. Unable to go to war on behalf of a Catholic, Elizabeth instead promises a safe exile in England as long as Mary does not aid her enemies. Mary responds that if she does, it will only be because Elizabeth forced her to do so, and threatens that should Elizabeth murder her, she should remember that she "murdered her own sister and queen". Placing Mary under house arrest, Elizabeth receives compelling evidence that Mary conspired with her enemies to have her assassinated, and orders Mary's execution. As Mary is walked to the scaffold, a remorseful Elizabeth cries for Mary, who reveals a bright red dress, implying herself a martyr. In her final thoughts, Mary wishes her son James well and hopes for peace upon his reign.
A post-script reveals that upon Elizabeth's death in 1603, James became the first monarch to rule both Scotland and England.
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