The Ligue 1 match between Nice and Marseille on Wednesday was paused after homophobic banners were displayed.
Two signs including homophobic language were seen in the crowd at Allianz Riviera, leading referee Clement Turpin to stop the game.
A delay of around five minutes followed before the match got back under way, with Nice player Wylan Cyprien critical of the interruption.
“We can’t stop a game every time that stupid people act like that,” he said to Canal+. “I am against every discrimination, homophobia or racism.
“But we can’t stop games for that. It’s ridiculous.”
Referees in France have been instructed to stop matches if homophobic banners are displayed, or homophobic chanting is heard, during the 2019-20 season.
The Ligue 2 match between Nancy and Le Mans on August 16 was the first to be temporarily suspended, while a referee threatened to do the same during the meeting of Monaco and Nimes in the top flight on Sunday.
The controversy at Nice came in the week British billionaire Jim Ratcliffe completed his long-awaited takeover.
Ratcliffe is the president of petrochemicals company INEOS, which this year bought the cycling outfit formerly known as Team Sky.
INEOS already owns Swiss Challenge League side Lausanne Sport, which it acquired in November 2017, and the company will remain on as Nice’s main sponsor for the 2019-20 season.
The club lost their first match of the campaign on Wednesday, with Marseille finishing as 2-1 winners away from home.
Marseille originally took the lead in the first half on a goal from Dario Benedetto before Cyprien levelled the match from the spot in the 66th minute.
However, a Dimitri Payet penalty in the 73rd minute sealed the match for Marseille, who picked up the club’s first win of the season having amassed one point from their first two matches.
Next up for Nice is a visit to Rennes on Sunday while Marseille visit Saint-Etienne on Sunday as well.