to lose in the second round, falling 2-6 6-3 6-4 to Anna Schmiedlova of Slovakia.
The 29th seed, wearing long sleeves in temperatures of 14 Celsius, has never won the claycourt grand slam in 16 visits and, now 32, she may have to accept she never will.
Schmiedlova, aged 19 and ranked 56 in the world, wasted two match points on Williams’s serve before the American hit wide to set up a third, and the Slovak clinched the match with a fine backhand cross court shot.
“This is the biggest win of my life,” said Schmiedlova, who broke down when expressing how much Venus had meant to her when she was growing up.
Venus was diagnosed with an auto-immune disease in September 2011 and since losing in the fourth round of Wimbledon that year, has exited in the first or second round of every grand slam she has taken part in.
“I think she just played so well and just kept getting so many balls in the court,” said Venus. “It just wasn’t my day in the last couple of sets.”
Flavia Pennetta of Italy, seeded 12th, joined the Williams sisters in heading for the exit after going down 5-7 6-4 6-2 to Sweden’s Johanna Larsson.
Eugenie Bouchard, the 18th seed, recovered from a slow start to beat Julia Goerges of Germany in three sets.
On the men’s side, Russian Mikhail Youzhny, the 15th seed, lost 6-0 6-3 3-6 6-4 to Czech Radek Stepanek before Spain’s Marcel Granollers overturned a two-set deficit to beat 20th seed Alexandr Dologopolov of Ukraine 1-6 3-6 6-3 6-0 6-2.
Gilles Simon, the 29th seed, eased past Alejandro Gonzales of Colombia 6-4 6-0 6-2 and there were also straight-set wins for Spain’s Roberto Bautista Agut, the 27th seed, and Ernests Gulbis, the 18th seed from Latvia.