Twilight Trail first starts off as an grassland; its colors are orange and brown. Twilight Trail takes place during nightfall. When Mario reaches the forest area of Twilight Trail, it turns dark and the area's color is blue and purple.
Mario and his friends had to venture through Twilight Trail several times throughout Chapter 4. This is mainly because Mario had his identity stolen by Doopliss and Mario had to go back and forth through Twilight Trail to chase Doopliss and find the secret to his name.