In this second 'Dukes of Hazzard' reunion film, the opening scene shows with Bo and Luke Duke partaking in the Hazzard County festival which features actual country music star Toby Keith as himself to raise money for a new hospital to be built in Hazzard. When the musical number is over, an old enemy of the deceased J.D. Hogg, Ezra Bushmaster, arrives. He offers to donate a significant amount of money ($3 million) to the hospital fund, on the condition that the people of Hazzard are able to raise the rest ($500,000) by a certain amount of time. Just then, a man who introduces himself as B.B. Bascomb, a music promoter on vacation in Hazzard, says he'll buy the master tapes of the performances of music stars who performed in Hazzard. So the Dukes decide to travel to Hollywood to sell some country music recordings to raise the rest of the money.
They are joined by Sheriff Rosco P. Coltrane in his RV accompanied by a fake basset hound named Einstein. Cooter Davenport and Cletus Hogg follow in a truck carrying Daisy's Motorcycle. On the way there they are shot at by a one eyed hitman, named Patch, who was hired by Ezra to stop the Dukes from arriving in California.
Once they get to Hollywood, they set up a camp in Hollywood Hills. Shortly thereafter, Enos Strate, now a full detective of the LAPD, joins them but informs them they need permits to camp next to the Hollywood Sign.
The next day, as Enos, Daisy, and Cooter drive to downtown L.A.'s city hall to get the permits, Patch and his partner steal the RV... along with all of the raised money and audio tapes as well. In an attempt to recover the RV and the music that was inside it, Bo and Luke save a Mexican teenager from a group of gang members. Soon, the boy's older brother arrives with friends in a lowrider convertible. They thank Bo and Luke and, after seeing what they can do with The General Lee and a ramp, invite them to a block party where Bo falls in love with Gabriela, a cousin of Cipriano (the older brother).
Meanwhile, Enos, Daisy, and Cooter arrive at city hall where it turns out that the deputy mayor, is in fact an old love interest in Enos since he arrived back in L.A. whom she owes a favor. She grants the camping permits and implies to Enos that she will demand a favor in the future. After leaving city hall, Enos, while driving Daisy and Cooter back to the campsite is called to settle a hostage situation where an armed man has taken a person hostage in a store. Daisy also sees that Enos knows an attractive female uniformed officer, named Tami, and knows the holdup man as an old school friend from Hazzard whom he talks into surrendering.
The next morning, Bo and Luke get information that the RV is at a chop shop. By the time they get there, the RV has been chopped for parts. But the music recordings and the money were not inside, so they continue looking when The General Lee's transmission goes out. Just then, they are confronted by a black woman, named Miss Deacon, who threatens to take their car which she thinks has a racing engine she wants, which they don't have.
After being confronted by the bag of rattlesnakes on The General Lee's seat, she asks them, "You boys aren't from New Jersey, are you?" It is revealed that the two hitmen that stole the RV went to her, and she threatened them the same way, and they told her that the boys had the racing engine she was looking for. She fixes The General Lee in exchange for the recipe to the late Uncle Jesse's old barbecue sauce.
In the meantime, Daisy gets work as a stuntwoman on a Hollywood film set, and Rosco meets up with an unfriendly loan shark, named Sharon, to try to get his money back in a loan, but the hitman, Patch, steals the money to deliver it to a local Russian mobster, named Igor the Terrible, who wants the audio tapes that Patch has in order to make bootlegged tapes for his distribution empire.
When Sharon and her henchmen arrive at the Duke's camp and threaten to kill Rosco to demand their money back, Enos arrives and it turns out that Sharon is in fact another old love interest in Enos whom he talks to into leaving out of fear of her parole being revoked. It is here were after Enos learns about the music promoter, that he explains that B.B. Bascomb was a big old fraud that had tangled with Enos in the past. Bo and Luke, aided by Enos, break into Bascomb's house who tells them about Igor the Terrible.
The boys then stake out the nightclub here Igor and Patch frequent where they attempt to steal back the tapes, only to get caught trying to get away. They are taken to a warehouse where Igor plots to kill both of them, but the day is saved when Gabriela, who had tagged along with Bo and Luke, calls her cousin's gang who assault the warehouse and defeat all of the Igor's men in a gunfight. Bo and Luke then capture Patch as he tries to escape with the tapes and Enos arrives with the police to arrest him.
With everything resolved, Bo and Luke and the rest of the gang return to Hazzard County and celebrate the groundbreaking of the new Hazzard Hospital. Gabriela and her family also show up for the festivities. But it is here were the boys finally discover that Ezra Bushmaster, the man who donated the money for the hospital, was the same man behind the theft all along. But with no evidence to arrest him, Ezra tells Rosco that he plans to run against him for city commissioner in the next election and will be the new 'Boss' of Hazzard. But he neverless shows that he still has a heart by allowing the festivities to proceed, as Bo and Luke and the rest of the gang realizes that they will now have a new nemesis to deal with in the future as the film comes to a close.