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Jungle Land is a vast jungle located somewhere on Mushroom World. Miles of tropical trees make up the primary atmosphere of Jungle Land, along with rivers, ponds, and the occasional quicksand pit. Animals that live in Jungle Land include apes, lions, Shy Guy Lions, Albatosses, Trouters and Fryguys. The ape couple June and Ward are two notable residents who live in a society populated by anthropomorphic primates. Fabled treasure hunter Indiana Joe also resides in another part of Jungle Land, where he lives in a hut built over small pond. Jungle Land is also home to the dreaded Temple of Koopa.
Mario, Luigi, Princess Toadstool and Toad initially travel through Jungle Land as means of a shortcut, but are soon ambushed by King Koopa and an Albatoss, which kidnaps Mario. Mario manages to escape from the Albatoss' grasp, and finds himself landing headfirst on the front porch of June and Ward's treehouse, causing him to lose his memory. June and Ward choose to raise Mario as their own child, rename him to "Marzan", and teach him how to behave like an ape. After mingling with some of the local monkeys, Mario beats up a lion, and proclaims himself as "Marzan, King of the Jungle".
In their attempt to rescue Mario, King Koopa orders his Shy Guys to attack Luigi, Princess Toadstool and Toad. Their attacks lure the heroes into a river, where they are almost devoured by a school of hungry Trouters. Mario swings down on a vine and rescues them, but fails to recognize his friends and leaves to return to his adoptive parents' home for dinner.
Luigi, Princess Toadstool and Toad follow Mario's trail of banana peels to his home, and proceed to climb up after him. Before they can restore Mario's memory, King Koopa orders his Shy Guys to chop down the entire tree, but Mario single-handedly beats them with his jungle ethics. Luigi inadvertently restores Mario's memory right after, but he is still able to finish his duty and send King Koopa and his men away from the jungle.
Ages ago, King Koopa's great-great grandkoop travelled to Jungle Land, where he discovered the wish-fufilling Lost Mushroom. What he used the Lost Mushroom for remains unknown, but according to King Koopa, it was supposedly left to him. Before dying, his Great-Great Grandkoop built the Temple of Koopa to hide the Lost Mushroom from the rest of the world, where it remained for many years.
Treasure hunter Indiana Joe moved his base of operations to Jungle Land to search for lost treasure of all sorts, and soon learned of the location of each and everyone one of them, in addition to all shortcuts leading to them. However, many of these treasures have yet to be found.
One day, Mario, Luigi, Princess Toadstool and Toad return to Jungle Land in a hot air balloon, as they are hoping to get ahold of the Lost Mushroom before King Koopa does. After sabotage from an Albatoss, they soon find themselves sinking in a quicksand pond. Indiana Joe rescues them, and offers to lead them to the Lost Mushroom.
Kolonel von Koop, not knowing the location of the treasure, lets Indiana Joe lead the way and steals it once it is found, locking Mario and his friends up in a tree hut with a Shy Guy Lion afterwards.
After taming the Shy Guy Lion, the group finds Indiana Joe at home and convince him to help them fight Kolonel von Koop despite his Koopaphobia.
In the meantime, Kolonel von Koop uses the Lost Mushroom to wish for a castle made of solid gold, but it is soon stolen back by Mario. Accompanied by Indiana Joe, Mario's group fights off against and defeats Kolonel von Koop and the Koopa Pack. With Kolonel von Koop gone from Jungle Land for good, its citizens can now live in peace.