When the Mario Bros. reach the ruined Castle Town, they immediately go to the castle, where they meet Lady Lima. She tells them to fix the castle's plumbing. When they do, the Beanstar's protective barrier vanishes and "Lady Lima" reveals herself to be Cackletta. When the Mario Bros. reach the room, she forces Queen Bean to attack them and leaves before the fight.
Her next appearance is in Woohoo Hooniversity, where she apparently turned all the scientists into monsters. After Peach's voice fails to awaken the Beanstar (because, as it turns out, the voice that Cackletta had stolen at the start of the game was actually that of Birdo's), the Mario Bros. come in and defeat her. Fawful sucks her spirit up with his headgear and prepares to blast Mario and Luigi. Prince Peasley then knocks Fawful and Cackletta's spirit to Stardust Fields.
There, a battered and beaten Fawful finds Bowser's unconscious body, and allows Cackletta to possess him. She then becomes Bowletta, a rather frightening hybrid of Bowser and Cackletta herself. She then takes control of both Beanbean and Koopa troops in a final battle against the Mario Brothers. After an initial battle with Bowletta, the Mario Brothers are taken inside of Bowletta, to confront Cackletta's angry spirit. After the spirit's defeat, Cackletta is defeated once and for all.