- Mario - He is our favourite hero and a playable character from the start. He has ballanced skills overall. In his small form, he hasn't got his hat.
- Luigi - Mario's little brother is also playable from the start and has better jumping skills. He is the best jumper! In his small form, he hasn't got his hat.
- Birdo - She is also playable from the start. She can suck characters by pressing Y, then throw them by pressing Y again.
- Toad - Toad, a mushroom guy, is an unlockable character. He has good running skills, but lacks power and doesn't jump very high. By pressing Y, you can roll with his mushroom cap. Unlock him by defeating Petey Piranha in Pink Hills. He'll be surprised and join the party.
- Daisy - The tomboy princess is also a playable character, but she is unlockable. She has ballanced stats like Mario, even though, she lacks running skills. In her small form, she hasn't her crown on. Unlock her by defeating Pom Pom in Power Airship. She'll be surprised and join the party.
- Toadette - Toad's younger sister also joins Mario and his crew to restore peace to Mushroom World. Her stats are like Toad's. Curiously, she has better jumping skills. Her Y move is also the same as Toad's. Unlock her by finding her in King Boo's Haunted Hideway. She'll be grateful and join the party.
- Princess Peach - Peach was not kidnapped in this game. She is a bit worried about the darkness around her castle and asks Mario and his crew to find the Power Stars. She gives the player a item if they find her around the castle, but just once.
- Toads - The Toads host Toad houses, where they sell items. They might also give the player items and useful information. They are found on the areas and in the levels.
- Piantas - Piantas give the player information. They are found around the areas and levels.
- Nokis - Nokis are just like Piantas. They are found around the areas and levels. However, there is a pink Pianta who lost her guitar in the Crystal Lake level.
- Lubba - He is on various levels, but most notably Moonview Hotel, were he is searching for the other Lumas.
- Red Lakitu - He only appears in the level, Shooting Star Metropolis, searching for his cloud, stolen by Goomboss.
- Toadiko - She appears next to Toadette after she is found, on the areas. When you talk to Toadette, and she joins the party, Toadiko goes out. On Ukiki Hills, she is looking for a pink Ukiki.
- Lumas - Appear in the levels of space-theme, as informating characters. In Moonview Hotel, a bunch have been kidnapped by Mouser.
- Balloon Cheeper
- Petey Piranha
- Whomp King
- Boom Boom
- Big Bob-omb
- Pom Pom
- Gooper Blooper
- Bowser Jr.
- Rocket Koopa
- King Boo
- Giga Dry Bones
- Dark Mario
- Ukiki King
There are various different types of enemies:
- Standard - Almost everywhere
- Flying - Mostly Athletic levels
- Weapon - Almost everywhere
- Ice - Only ice-themed
- Floral - Almost everywhere
- Underwater - Only underwater levels
- Cave - Mostly cave levels
- Spooky - Mostly mansion/castle levels
- Castle - Mostly castle levels
||Your character is small and has 5 points of health. You revert back to it if you lose 5 points of health when powered-up. You can't break brick blocks nor spin attack.|
||Your character gets taller and has 10 points of health. If you lose 5 of them, you revert back to small character. You can break brick blocks and spin attack.|
||Your character gains the ability to shoot Fireballs. His/her clother might turn red or his/her colour will change. You can kill most enemies with fireballs. If you get hit 1 time, you revert back to Super character.|
||Your character gains the ability to shoot iceballs, that freeze the opponents for 5 seconds. If you touch them, they will die. If you get hit 1 time, you revert back to Super character.|
||Your character turns into Starman, being invincible from any attack. However, Starman may still fall off cliffs. This power-up is temporary.|
|Your character turns giant, destroying everything in your way. You still need to jump and it may fall off oversized cliffs. This power-up is temporary.|
|Your character gains the ability to shoot hammers. It starts with a amount of 10 hammers. If you find another Hammer Suit, you might get more 10. If you get hit 1 time, you revert back to Super Character.|
Your character gains the ability to shoot boomerangs. It starts with a amount of 10 boomerangs. If you find another Boomerang Suit, you might get more 10. If you get hit 1 time, you revert back to Super Character.
Your character is able to attack enemies with its tail by pressing B. You can also fly for a limited time by holding A while in mid-air. If you get hit 1 time, you revert back to Super Character.