Serena Williams' bid for the Italian Open was squashed in her first match on Tuesday when the American second seed lost 6-2, 2-6, 6-1 to Swiss Patty Schnyder.
Williams appeared out of sorts from the early stages of her match, which started after 2300 local (2100 GMT) because of rain delays and the length of the preceding matches.
She dropped serve twice in the first set, the second time to love, then came back strongly in the second set only to slump out in the decider.
Champion Jelena Jankovic repelled a gutsy challenge from Argentina's Gisela Dulko to move into the third round with a 6-3, 7-6 win and world number one Dinara Safina also had to work hard to beat France's Virginie Razzano 7-6, 6-1.
Jankovic, who has slipped to number four in the rankings after ending 2008 top, had to save four set points to hold serve for 5-5 in the second set before a storm halted play for three and half hours.
The Serb, who is seeking her third consecutive title in Rome, wasted four match points in the tiebreak before clinching it 8-6 to book a clash with Kateryna Bondarenko, who knocked out French 13th seed Alize Cornet, last year's runner-up, 6-4, 6-1.
"I complicated my life out there," Jankovic told a news conference. "I had everything under control and all of a sudden she's on top of me and she has set points.
"It was difficult but I managed to somehow stay positive and stay strong and win that second set."
Razzano's power and clever use of the drop shot on the damp clay caused plenty of problems for Safina.
BREAK POINTS
The Russian twice had to recover the loss of her serve in the first set and saved three break points in the 11th game before winning the tiebreak 7-1.
Safina then ran away with the second to set up a clash with Chinese 14th-seed Zheng Jie.
The top seed took a nasty tumble in the first set and was not happy about the wet surface.
"I was just so angry they made us play on this court because it's not perfect," said Safina. "You can get injured. It's not funny.
"Thank God I didn't break anything. They have to have a little respect for us players."
French Open champion Ana Ivanovic beat Italian Francesca Schiavone 6-3, 6-4 and seventh seed Svetlana Kuznetsova of Russia had a comfortable 6-3, 6-3 win over Slovak Daniela Hantuchova.
