The Australian, who won the women’s singles title in Paris in 2019, lost the first set 6-1 and was level at 2-2 in the second before deciding to retire with a left hip issue.

"It’s heartbreaking," said Barty who struggled with the injury during her first round victory over American Bernarda Pera.

"I mean, we have had such a brilliant clay court season, and to kind of get a little bit unlucky with timing more than anything to have something kind of acute happen over the weekend and just kind of run out of time against the clock is disappointing.

"It won't take away the brilliant three months that we have had, as much as it hurts right now."

While Barty bowed out, Poland’s Iga Świątek underlined her ambitions to win a second successive French Open crown with a 6-1, 6-1 thrashing of Sweden’s Rebecca Peterson.

Sofia Kenin of the United States, who lost to Świątek in last year’s final, was forced to play on Court 14 but produced a professional performance to beat compatriot Hailey Baptiste, a qualifier who won four matches in Paris in her French Open debut, 7-5, 6-3.

Kenin will now face another American in Jessica Pegula, who eased to a 6-3, 6-3 win over Tereza Martincova of the Czech Republic.

Fifth seed Elina Svitolina of Ukraine also cruised through to the third round, defeating Ann Li of the US 6-0, 6-4 in one hour and 14 minutes, while Tunisia’s 25th seed Ons Jabeur thumped Australian wildcard Astra Sharma 6-2, 6-4.

Seeded ninth, former world number one Karolína Plíšková of the Czech Republic exited the tournament following a 7-5, 6-1 loss to American Sloane Stephens.

Either Marta Kostyuk of Ukraine or Varvara Gracheva of Russia will reach the second week of a Grand Slam for the first round after the youthful pair set up a third round meeting.

Kostyuk, 18, backed up her first round upset of Spanish 12th seed Garbiñe Muguruza with a 6-3, 6-4 win over Zheng Saisai of China, while Gracheva, 20, overcame Camila Giorgi 7-5, 1-6, 6-2 to book her place in the third round of a major for the second time, following the 2020 US Open.

American teenager Coco Gauff’s superb clay court season continued with a 6-3, 7-6 (7-1) success against Wang Qiang of China.

Gauff will next become involved in an all-American showdown in the third round where she will take on 2021 Australian Open runner-up Jennifer Brady who outlasted the top-ranked Frenchwoman Fiona Ferro, 6-4, 2-6, 7-5, in two hours and 11 minutes.

There were also victories for Estonia’s Anett Kontaveit, Belgium’s Elise Mertens, Greece’s Maria Sakkari and Czech pair Barbora Krejčíková and Karolína Muchová.

In the men’s draw, Spain's Rafael Nadal, who is hunting a 14th Roland Garros title, and a fifth in a row, was involved in the night match on Court Philippe-Chatrier.

Nadal produced a masterful display as he overpowered France's Richard Gasquet 6-0, 7-5, 6-2.

Twenty time Grand Slam champion Roger Federer found another level to battle past former world number three Marin Cilic on Court Philippe-Chatrier.

The Swiss great prevailed 6-2, 2-6, 7-6 (7-4), 6-2 against the 2014 US Open champion from Croatia to move through to the next round.

Federer, who landed in Novak Djokovic’s half of the draw, will next face Germany’s Dominik Koepfer who defeated American 30th seed Taylor Fritz 6-3, 6-2, 3-6, 6-4.

Serbia’s Djokovic beat Uruguayan clay court specialist Pablo Cuevas 6-3, 6-2, 6-4 to continue his quest for a first Roland Garros title since 2016.

He will next play Lithuania's Ricardas Berankis, who beat James Duckworth of Australia 7-5, 2-6, 7-6 (7-4), 6-0.

"I'm ready to go deep in this tournament," Djokovic said.

"Hopefully that's going to be the case."

Matteo Berrettini of Italy completed his 20th match win of the year for a place in the Roland Garros third round for the third time as he saw off Argentina’s Federico Coria 6-3, 6-3, 6-2.

Italians Jannik Sinner, Lorenzo Musetti and Marco Cecchinato joined compatriots Berrettini and Fabio Fognini in the third round.

Sinner defeated another Italian in Gianluca Mager 6-1, 7-5, 3-6, 6-2, Musetti struck 39 winners, including four aces, against Yoshihito Nishioka of Japan in a 7-5, 6-3, 6-2 victory and Cecchinato beat Alex de Minaur of Australia 6-4, 6-1, 3-6, 6-1.

Sweden’s Mikael Ymer downed French 14th seed Gael Monfils 6-0, 2-6, 6-4, 6-3, while promising Spanish player Carlos Alcaraz ousted Georgian 28th seed Nikoloz Basilashvili 6-4, 6-2, 6-4.

There were also wins for Argentine 10th seed Diego Schwartzman, Britain’s Cameron Norrie, South Korean Kwon Soon-woo and German duo Jan-Lennard Struff and Philipp Kohlschreiber.