Cousins Bo, Luke, and Daisy Duke run a moonshine business for their Uncle Jesse in Hazzard County, Georgia. The cousins' primary mode of transportation is an orange 1969 Dodge Charger that the boys affectionately refer to as the "General Lee". Along the way, the family is tormented by corrupt Hazzard County Commissioner Jefferson Davis Hogg, widely known as "Boss Hogg", and his willing but dimwitted henchman, Sheriff Rosco P. Coltrane.
After Rosco has the General Lee vandalized after Bo and Luke's attempt to run away from a daughter of one of their many moonshine customers, Billy Prickett, a famous stock-car driver, enters Hazzard to participate in the rally. Meanwhile, Rosco plants a fake moonshine still in Uncle Jesse's barn and seizes the Duke property in the interest of eminent domain for Boss Hogg, forcing the family to temporarily reside with neighbor Pauline. She informs the Dukes that Rosco seized another farm on charges, so Bo and Luke investigate a local construction site and find their farm and neighboring farms marked on a map. They also discover a safe which they believe contains information that Boss Hogg wants to keep secret, unable to get the safe open they drag it to Sheev's. With his help they blow open the safe and discover geologic core samples but are unsure on what the samples are. Meanwhile, Cooter turns the General Lee into a hot rod in hopes of finally receiving payment for all the work he has done for the boys in the past if they win the race.
The Dukes go to Atlanta to visit a local university geology lab, meeting with Katie-Lynn Johnson, a Hazzard county girl. They discover Boss Hogg's intentions of turning the county into a strip coal mine. However, before they can return to Hazzard country, they are arrested. Boss Hogg visits their prison cell, where he informs the Dukes that the vote on Hogg's proposition is at the same time as the rally, with Billy Prickett providing an distraction. During a transfer from detainment, Daisy helps the boys escape from the patrol car, and they speed home to try to inform the townsfolk, escaping the Atlanta Police, and the Georgia State Patrol.
Upon returning home, the Dukes discover that Boss Hogg and Rosco had taken Uncle Jesse and Pauline hostage. The two race to the farmhouse to cause a distraction to the waiting Hazzard County Sheriff's deputies and Georgia State Troopers, while Daisy and Cooter rescue Jesse and Pauline. Meanwhile, the college girls head to the rally with Sheev to inform the townsfolk about the vote on the strip-mining ordinance, however; because Sheev is a known conspiracy theorist, no one listens to them. Bo leaves for the rally while Luke and Jesse team up to foil the county and state police. Upon crossing the finish line first before Billy, the two then continue racing across town, leading the townsfolk to the courthouse, where the proposal is rejected.
Boss Hogg attempt's to have the boys locked up but Daisy takes advantage of the governor of Georgia's presence and TV cameras to convince him to pardoning them. The final scene shows a cook-out at the Dukes' house where the Dukes and the townspeople celebrate.
Johnny Knoxville as Luke Duke
Seann William Scott as Bo Duke
Jessica Simpson as Daisy Duke
Burt Reynolds as Boss Hogg
Willie Nelson as Uncle Jesse Duke
David Koechner as Cooter Davenport
M. C. Gainey as Sheriff Rosco P. Coltrane
Michael Weston as Deputy Enos Strate
Lynda Carter as Pauline Powers
James Roday Rodriguez as Billy Prickett
Kevin Heffernan as Derek "Sheev" Sheevington
Nikki Griffin as Katie-Lynn Johnson
Jacqui Maxwell as Annette[2]
Alice Greczyn as Laurie Pullman
Junior Brown as The Balladeer (narrator)
Joe Don Baker as Governor Jim Applewhite
Barry Corbin as Bill Pullman
Andrew Prine as Angry Man
Brendan Schetter as Random Stoner
Michael Roof as Dil Driscoll
loosely based on the television series of the same name.
The film was the acting debut of pop singer Jessica Simpson.