The game focuses on the first ever Mater-National race held in Radiator Springs by Lightning McQueen and features an improved world as well as new characters. New characters featured are Emma, a small white and pink rally car from England, Gudmund, a cocky 1980 Audi Quattro rally car from Sweden who has loads of headlights and loves driving through caves at night, Otto, a German tuner car who doesn't understand the concept of speed limits, Koji, a red, grey and white Japanese drift car who loves drifting, Yuri, (possibly named after Yuri Gagarin), a dark green Russian 1962 Toyota Crown, Philip, a red and white, over-excited British 1937 Cord Model 812 who has trouble remembering which side of the road to drive on in America, and Giovanni, a tough-looking but very nice red Ferrari Enzo from Italy. Ornament Valley, Radiator Springs, and Tailfin Pass have all been updated. In Radiator Springs, new rock formations have been cut, and Fillmore has opened a nature park. In Ornament Valley, there is a construction zone, an airport, and several new rock formations to explore. In Tailfin Pass, roads have been completely revamped, and there is a new mine and an abandoned western town to drive through. But some roads are blocked off and can only be played in races and as a result to this, the three areas are smaller than in Cars: The Video Game.
Before the launch, they were trying to copy all of the roads from Cars: The Video Game into Mater-National plus the newly designed roads for the three areas of Carburetor County. Unfortunately, there were some problems, if all was included the frame rate would be close to about 20 or 40. The only way to unlock the roads is to at least see to win and get all bolt banners in the game. If that isn't the case, try some cheats, if not, forget about it. Other issues where that for some reason [the issue is a good issue] you can drive around the areas as Monster McQueen which is faster than the regular McQueen. Other things, well the game has very little races and each unlocked area is a story mode. Sadly, there is only 4. The Stadium in Mater National is actually the Radiator Springs Raceway and for the first time ever, Chick Hicks was not in the game. Also Mater-National has border issues that don't connect but are shorter because of the designing issue. Other than that, that's all the issues.
- Radiator Springs Circuit
- Fillmore's Nature Preserve
- North Willy's Butte
- Ornament Valley Airport
- Inside the Turkey
- Rustbucket Grand Prix
- Wheel Well Circuit
- The Upper Mine
- Canyon Run
Luigi and Guido's Team Relay
- Luigi and Guido's Team Relay 1
- Luigi and Guido's Team Relay 2
- Luigi and Guido's Team Relay 3
- Luigi and Guido's Team Relay 4
Monster Truck Waypoint Races
- Mike and Sulley's Last Laugh (6 levels)
- Fillmore's Fuel Frenzy (9 levels)
- Doc and the Law's Race 'n' Chase (9 levels)
- Ghosting Mater (7 levels)
- Ramone's Rhythmic Rumble (7 levels)
- Tractor Tipping (6 levels)
- Snot Rod is the only one of the Delinquent Road Hazards to appear in the game.
- Although E.J. Holowicki returned for the game, DJ doesn't appear.
- Similar to Cars: The Video Game, the Radiator Springs Theme is used for the main menu.
- Unlike Cars: The Video Game and Cars Race-O-Rama, music doesn't play in exploration mode, except for when the player goes near an event.
- On the DS version, Lightning has a phrase, "Ka-chow" when he jumps, but it sounds as if it was done by a child.
- If Otto, Gudmund, Emma, Koji, and Giovanni were big champions in their home countries, they should likely have been in the World Grand Prix in Cars 2, but none of them did.
- In the Rustbucket Grand Prix end-scene, Gudmund and Mater ran away from each other. Mater's headlight stopped moving right after he ran away.
- In Cars Mater-National, he said, "Konichiwa!", the Japanese word for "Hello" and "Good afternoon". But in the Cars 2 In Flight to Japan video, when a forklift on the television screen in the plane said the word, Mater misheard her, saying, "Konichi-what?", as if he hasn't heard of the word.
- On the GameBoy Advance version, several signs that are supposed to be U.S. Route 66 signs are shown as U.S. Route 99 signs.