James Stewart, Duke of Rothesay (22 May 1540 – 21 April 1541) was a short-lived heir to the throne of the Kingdom of Scotland.He was the eldest son of James V and Mary of Guise, and nephew of his namesake James, Duke of Rothesay. At the time of his birth in St Andrews, James V had survived his own brothers. The newborn Duke of Rothesay and his father were the only living legitimate descendants of his paternal grandfather James IV of Scotland. The heir presumptive since 1536 was his second cousin James Hamilton, 2nd Earl of Arran.