Mike Gillislee ran for a pair of second- half touchdowns to lead No. 10 Florida past fourth-ranked LSU, 14-6.
Gillislee finished with 146 yards on 34 carries for the Gators (5-0, 4-0 SEC), who racked up 176 yards on the ground to remain undefeated.
Jeff Driskel completed 8-of-12 passes for 61 yards and was backed by a stifling defense that held LSU to 1-for-13 on third-down conversions.
Zach Mettenberger went 11-for-25 through the air for 161 yards and an interception for the Tigers (5-1, 1-1), who had their 18-game regular-season win streak snapped.
LSU had not dropped a game in the regular season since a 31-23 decision to Arkansas in the 2010 finale.
The Tigers' Odell Beckham paced all receivers with 81 yards on four receptions in defeat.
"We beat an outstanding football team. They have won 18 straight regular season games for a reason," Florida coach Will Muschamp said. "You saw two very physical teams go at it today on both lines of scrimmage and you could see the improvements that we have made."
LSU received the ball to start the game and quickly moved down the field to take the early lead. Mettenberger had completions of 20 and 13 yards to Spencer Ware and Nic Jacobs, respectively, before Kenny Hilliard reeled off a 13-yard rush to move the ball deep into Florida territory.
But the Tigers would not record another first down and Drew Alleman capped the 10-play march with a 31-yard field goal.
The teams traded punts from there until Jaylen Watkins made a leaping interception of an errant Mettenberger pass on LSU's first touch of the second quarter, giving Florida the ball at the Tigers' 38.
Florida would not capitalize on the turnover, as after Gillislee's 5-yard run on first down, back-to-back sacks by Kevin Minter pushed the Gators out of field-goal range and they were forced to punt.
The Gators regained possession following an LSU punt and drove the ball down to the Tigers' 22-yard line, but Frankie Hammond Jr. had the ball punched out of his hands during a short reception and LSU's Kwon Alexander recovered the fumble.
The Tigers were held to a three-and-out on the ensuing drive, but later used another Florida turnover to add to their lead when Driskel fumbled during a sack and Barkevious Mingo jumped on ball at the Gators' seven, resulting in Alleman's 21-yard field goal for a 6-0 advantage at the break.
Florida got on the board on its second possession of the third quarter with an 8-play, 85-yard scoring drive that did not include a third down.
Gillislee carried the ball four times for 29 yards on the march, including a 12-yard touchdown on a run to the right side for a 7-6 lead with 5:15 left in the third.
The Gators had managed just 49 yards of offense prior to their go-ahead drive.
LSU looked to quickly answer as Mettenberger connected with Beckham three plays later on a deep pass down the right sideline for a 56-yard gain, but Matt Elam stripped the ball away from Beckham on the play and Florida recovered at its own 23-yard line.
Florida took advantage of the turnover with an 11-play drive consisting entirely of runs and was finished off with Gillislee's 12-yard score with 13:18 remaining in the game.
The Tigers would not threaten down the stretch, as their offense recorded just two first downs and did not advance the ball further than their own 43-yard line.
"The opportunities we had -- if we had kept possession on the reception in the second half -- certainly could have made a difference in the game," LSU coach Les Miles said. "With that being said, we will make corrections and understand where we are at. I think our football team is sick, sick with knowledge that they could have played better."
It was just the fourth time in the all-time series that each team entered the contest ranked in the top 10. Florida leads the series, 31-25-3 ... Ware finished with 21 yards on eight carries, and 20 yards on one catch ... LSU totaled just 42 rushing yards in the contest ... Florida went 7-for-14 on third-down conversions.