England 1536. Despite having given birth to the future Queen Elizabeth I, Anne Boleyn (Anna), the second wife of King Henry VIII (Enrico), has failed to give him a son and heir. His eye has strayed to Anne’s lady-in-waiting, Jane Seymour (Giovanna).
Scene 1: Queen’s Apartments
Courtiers discuss the state of royal affairs. Anna has summoned Giovanna and admits that she is troubled. She then asks her page Smeton to cheer people up with a song, but the song reminds Anna of her first love, which she betrayed to marry the king. Anna and the others depart, leaving Giovanna — who is the king’s new lover — alone and guilt-ridden. Enrico enters and declares his love for Giovanna, promising marriage.
Scene 2: A Park in Windsor
Anna’s brother, Lord Rochefort, is surprised to meet Richard Percy, whom the king has recalled from an exile imposed for being the queen’s former lover. Percy asks after Anna; Rochefort answers evasively.
Enrico arrives with a hunting party. Anna and her ladies-in-waiting follow. Enrico greets Percy coldly and tells him that Anna is responsible for his pardon. But in allowing Percy to return, Enrico has set a trap.
Scene 3: Near the Queen’s Apartments
Smeton, secretly in love with the queen, is coming to return a locket that he has stolen which contains Anna’s portrait. He hides when Anna and Rochefort appear. Rochefort wants Anna to persuade Percy to leave England and danger. Percy enters, still in love with Anna. Although she tells him that the king now loathes her, she urges him to leave England and when she refuses to see him again, he draws his sword to stab himself. Smeton, thinking Anna in danger, rushes out to confront Percy. Rochefort returns with a warning that the king is coming. Enrico bursts in. Smeton proclaims the queen’s innocence but the king snatches the locket as proof of his wife’s infidelity and has her arrested.
Scene 1: London, Antechamber of the Queen’s Apartments
Anna has been imprisoned in her apartments; the guards note that even Giovanna has avoided her. Anxiously awaiting news of the trial, Anna’s ladies tell her to place her trust in Heaven. The ladies are summoned to give evidence before the Council of Peers. Giovanna steals in to tell that Anna can only avoid death by admitting adultery, allowing the king to divorce her. Anna dismisses her, but reflects that the betrayal is Enrico’s, not Jane’s.
Scene 2: Outside the Council of Peers
Believing his confession will save her, Smeton has falsely confessed to being one of the queen’s lovers. Anna says that she is ready to die but begs to be spared the humiliation of a trial. Playing into the king’s hands, Percy in turn claims that he and Anna were married before she became the king’s wife. Both are led away.
Giovanna pleads for Anna’s life, but the Council’s verdict is that the royal marriage is dissolved and Anna and her accomplices are to be executed.
Scene 3: Tower of London
Percy and Rochefort are together in their cell. They resolve to die together with Anna. Anna, meanwhile, is delirious in her cell, imagining it is the day of her wedding to the king and her thoughts return to her first love for Percy, and finally her childhood at her family home. Her fellow prisoners are brought in. Smeton blames himself; Anna asks him why he is not playing his lute. At the sound of a cannon announcing the king’s new marriage, Anna regains her senses. Despite being the victim of a horrendous crime, Anna says she does not wish vengeance on Enrico and Giovanni and goes to face her execution.