The game opens with Diddy Kong standing on the deck of a pirate ship. In the ship's cabin, he finds a note stating that Donkey Kong has been kidnapped by Kaptain K. Rool, a pirate Kremling, who had demanded the entire Banana Hoard that he stole in the previous game for a ransom from the Kongs. Diddy Kong subsequently attempts to rescue Donkey Kong from the Kaptain, and accompanying him is his girlfriend, Dixie Kong. Together, they travel through Crocodile Isle and are helped on their way by an assortment of animals (called Animal Buddies) in a bid to defeat K. Rool and rescue Donkey Kong.
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