Also in action on the first day at Roland Garros is men's second seed Alexander Zverev who meets Ricardas Berankis of Lithuania on Court Suzanne-Lenglen.
By losing, Ostapenko joins a ignominious group including, as well as Myskina, Steffi Graf, Jennifer Capriati, Svetlana Kuznetsova and Angelique Kerber who all lost as Grand Slam champions on the first run of their defences.
Sixth seed Elise Mertens fell to Kaia Kanepi 0-6, 6-0, 7-5 and seventh seed Julia Goerges lost to American Taylor Townsend 6-4, 7-6(0).
Novak Djokovic brushed past France's Adrian Mannarino 6-4 6-3 7-6(5) on Wednesday to book his place in the third round of Wimbledon. The defending champion, aiming for his third consecutive title, played well within himself and was rarely troubled during the second-round match. Mannarino, ranked 55, put up spirited resistance in the third set to force a tiebreak but was ultimately swept off Centre Court by the Serbian's laser-guided ground-strokes.
It wasn't just the tennis that amazed audiences at Roland Garros.
World No 2 Rafael Nadal brushed aside American Jared Donaldson 6-1, 6-1 and will next face Diego Schwartzman after the Argentine beat Roberto Carballes 6-3, 6-1.
World number one Serena Williams opened her US Open account with a tidy 6-3, 6-3 win over Ekaterina Makarova on Tuesday, to join her sister Venus in the second round of the year's final Grand Slam.
American teenage sensation Coco Gauff became the youngest player to win a WTA title in 15 years as she beat former French Open champion Jelena Ostapenko in the Linz Open final on Sunday.
The French Open witnessed a stunning upset on its very first day as 18-year-old Anastasia Potapova outclassed Angelique Kerber 6-4, 6-2 in the first round match on Sunday.
Jelena Ostapenko became only the sixth female Grand Slam champion to lose in the opening round of their title defence on Sunday when she lost to Kateryna Kozlova at the French Open.
Images from Day 1 of the Australian Open, in Melbourne, on Monday.
Images from Day 3 of the 2020 US Open in New York, on Wednesday.
Rafael Nadal left German qualifier Yannick Maden no hope as he booked his ticket for the third round of the French Open in ruthless fashion with a 6-1, 6-2, 6-4 victory.
A summary of Tuesday's play at the French Open.
Images from Day 1 of the US Open in New York, on Monday.
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