Venus Williams followed Serena to the French Open exit door, falling to Agnieszka Radwanska in the second round.
Making her first Grand Slam appearance after being diagnosed with Sjogrens syndrome, Williams struggled through her first round to set up a meeting with the third seed, but was no match for her Polish opponent. Radwanska eased past Venus 6-2, 6-3. It was the second day in a row that a Williams lost in the French Open, after sister Serena was shocked by Virginie Razzano in the opening round on Tuesday.
Venus, who looked tired after the match, said: "I felt like I played. I wasn't at my best today, yeah.
"This tournament for me was all about getting to the Olympics, if that happens to me, I think the chances are good, then I come out OK.
"She played really well. She put the ball in the court and chased down a lot of shots, that's what you have to do on this surface."
Five other American women - Melanie Oudin, Vania King, Alexa Gatch, Bethanie Mattek-Sands and Irina Falconi -- also saw their Roland Garros campaign come to a premature end.
The only American to win was Sloane Stephens who beat Mattek-Sands 6-1, 6-1 in an All-American clash.
Number one seed Victoria Azarenka eased to the third round with a 6-1, 6-1 victory over Dinah Pfizemaier.
Home favorite and eighth seed Marion Bartoli, however, crashed out to Petra Matic 6-2, 3-6, 6-3.
In the mens draw, Roger Federer defeated Adrian Ungur 6-3, 6-2, 6-7 (6-8), 6-3 for a record 234th victory in Grand Slam matches. Federer had tied with Jimmy Connors for 233 wins after winning his first round match, but victory in the second round saw the Swiss master move ahead.
Novak Djokovic also had no problems, dispatching Blaz Kavcic 6-0, 6-4, 6-4.