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Mario lands in the Mushroom Kingdom, ruled by Princess Peach,
while Luigi lands in the Dark Lands, ruled by the evil Koopa king Bowser.
Bowser seeks to marry Peach and will destroy the Mushroom Kingdom using a Super Star if she refuses. He imprisons Luigi to threaten Mario, whom he sees as competition for Peach's love. Mario meets Toad, who takes him to Peach. Peach plans to ally with the primate Kongs to help repel Bowser and allows Mario and Toad to travel with her. In the Jungle Kingdom, King Cranky Kong agrees to help on the condition that Mario defeats his son, Donkey Kong, in a fight. Despite Donkey Kong's immense strength, Mario defeats him using a Cat Suit.

Mario, Peach, Toad, and the Kongs use karts to drive back to the Mushroom Kingdom, but Bowser's army ambushes them on Rainbow Road.