In the movie Luke is first seen laying in his hammock outside when his wife Ellie Mae comes out of the house and tells him to wake up because their friend Orville the Albatross is in trouble, to which Luke drinks his moonshine which wakes him up. As he is walking down the steps of his house he is accidentally knocked over by Orville. When Bernard and Bianca fall in the swampy water Ellie Mae helps Bianca while Luke helps Bernard and gives Bernard a swig of his moonshine. He then asks Bernard where he and Bianca are from to which Bernard says that they're from New York City. The 4 of them spot the evil Madame Medusa to which Luke and Ellie say that they and the rest of the Swamp Folks would like to run her clean out of their Bayou, They also spot the kidnapped little girl Penny who is being taken back to Medusa's boat by Medusa's two pet crocodiles Brutus and Nero. Luke and Ellie Mae have their friend Evinrude the dragonfly take Bernard and Bianca to the boat while they round up the neighbors.
Later on after rounding up all the Swamp Folks Ellie Mae spots Luke asleep and tells him to wake up to which Luke replies "I was just waiting for someone to holler CHARGE!"
Later on, when Evinrude finally returns and is worn out of breath from escaping a gang of hungry bats, Luke gives him a sip of his moonshine which makes Evinrude recover his strength and urges and the Swamp Folks to follow him to the River Boat. Luke then takes another drink of his moonshine and yells "CHARGE!" while joining the Swamp Folks. Once aboard the River Boat the Swamp Folks manage to fight off Medusa with Luke that stabbed her bottom with his pitchfork. After trapping Brutus and Nero in a cage, Luke, Ellie Mae, and the other Swamp Folks get on Medusa's Swampmobile with Penny, Bernard, and Bianca. When Bernard finds out that the engine won't start due to lack of gas, Ellie Mae tells Luke to fill up the engine with his moonshine. Luke reluctantly sprinkles a few drops until his wife forces him to pour the whole thing in, which finally gets the engine going. Medusa manages to grab a hold of the back of the rope of the Swampmobile but luckily, they manage to turn around causing the rope to snap and Medusa to run into the River Boat's smokestack. Luke, Ellie Mae, and the others then celebrate their victory while escaping the Swamp with Penny, Bernard, and Bianca on the Swampmobile.