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No. Isabella of Castile was member of the Trastamara line, as her husband Ferdinand of Aragon. Her daughter, Juana, was the last Trastamara member who ruled in Spain. After she, her son, Charles I of Spain and V of the Holly Roman Empire was the first Habsburg monarch of Spain. His father was Philip of Habsburg, known as Philip the Handsome, son of the emperor Maximilian and heir of Burgundy.

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Q: Is Isabella of Castile considered a member of the Habsburg line?
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