Amelie Mauresmo, seeded six, has pulled out of Wimbledon, her spokeswoman Marie-Claude Previtali said on Friday.
The Frenchwoman is struggling with a rib injury which is hindering her service.
Mauresmo reached the semi-finals last year and was due to play a qualifier in the first round next week.
Her place in the draw will be taken by Russia's Svetlana Kuznetsova, who has been seeded 33rd. Lucky loser Seda Noorlander of the Netherlands will fill Kuznetsova's spot.
Mauresmo, the French number one, missed the Australian Open in January because of a knee injury.
Doubts about Wimbledon had surfaced after she was beaten in the second round of the Ordina Open in Den Bosch on Wednesday.
Previtali told Reuters by telephone Mauresmo had undergone tests on Thursday before making her decision on Friday morning after discussion with coach Loic Courteau.
"Amelie is looking to resume training to be ready in time for the Fed Cup which takes place after Wimbledon," she said.
France have reached the Cup quarter-finals where they meet Spain on July 19-20.
Mauresmo won the Warsaw Cup event in early May, beating Venus Williams in the final where the American was forced to retire with stomach pains when trailing 3-0 in the final set.
{{/usCountry}}Mauresmo won the Warsaw Cup event in early May, beating Venus Williams in the final where the American was forced to retire with stomach pains when trailing 3-0 in the final set.
{{/usCountry}}The Frenchwoman went on to beat Serena Williams in the Rome Masters semi-finals only to lose to Belgium's Kim Clijsters in the final.
She then suffered desperately in front of her own crowd at Roland Garros when Serena took a 6-1 6-2 revenge in the French Open quarter-finals.