Czech Vondrousova, who became the first unseeded woman to lift the title last year, earned a far less welcome accolade after a 6-4, 6-2 defeat by Spain’s Jessica Bouzas Maneiro.
Odds will be stacked against the seven-time Wimbledon champion and the holder of 24 grand slam titles when the tournament gets under way following recent knee surgery.
Defending champion Carlos Alcaraz drawn to face top seed Jannik Sinner in the semi-finals while women’s top seed Iga Świątek is in same section as last year’s winner, Marketa Vondrousova.
Everything you need to know as tennis enters the grass court season – which means the greatest players flocking to London to dress in all white and play at the All England Club.
The 37-year-old Scot went under the knife on Saturday after withdrawing from his second-round match at the Queen’s warm-up event in London on Wednesday.
The former world number ones, plus local hope and US Open winner Emma Raducanu, were granted entry into the main draw after taking lengthy breaks from the sport.
The four-time grand slam winner, who lit the cauldron at opening of the 2021 Tokyo Games, returned to tennis late last year after a 15-month maternity break.
Spaniard, 21, claimed third grand slam with five-set victory over Alexander Zverev at Roland Garros which means he has now won a major on all three surfaces.
Carlos Alcaraz is a two-time grand slam winner, while Alexander Zverev is still seeking his first. However, this will be the first final at Roland Garros for either player.
Novak Djokovic withdrew from the French Open with an injured right knee on Tuesday, ending his title defence and meaning he will relinquish the No 1 ranking.
Big-hitting Belarusian won 6-2, 6-3 and has now dropped only 18 games over her four matches, while fourth-seed Elena Rybakina breezed past Elina Svitolina 6-4, 6-3.
Zhang, who made history in Paris 12 months ago, lost 6-3 6-3 6-1 to the Greek hitter on Friday with Tsitsipas winning the match in just 91 minutes to set up a meeting with Italian Matteo Arnaldi.
Goffin said a fan spat chewing gum as he defeated a French player, and follows acts of jeering this week and last year; French players say the fans ‘add spice’.
Nadal, the undisputed king of Roland Garros with 14 grand slam wins there, was beaten in straight sets in first round, raising suspicions it could have been his last appearance.
Zhang Zhizhen beats Australian Aleksandar Vukic 6-4, 4-6, 6-3, 7-5, while Wang Yafan downs Maria Timofeeva 6-3, 6-3. Naomi Osaka into second round and potential clash with Iga Świątek.