The 23-time Grand Slam champion defeated fellow American Kristie Ahn 7-6 (7-2), 6-0 on Monday, but organisers confirmed her withdrawal ahead of her scheduled second-round encounter on Wednesday morning.
In a press conference, Williams confirmed that she is set to take the rest of the year off in order to let her body recover, with the stresses put on herself during her recent run to the US Open semi-finals taking its toll.
The 39-year-old picked up an Achilles injury during her semi-final defeat against Victoria Azarenka at Flushing Meadows earlier this month, and although she did not speak of any issues after her first-round victory in Paris, a decision was taken to withdraw from Roland Garros in order to recover.
"After I warmed up - it was a very short warm-up - I talked to my coach and I was like, 'What do you think?'” Williams said.
“We both thought about it and we really realised that it likely wasn't the best for me to play today.
"I love playing in Paris, I really wanted to give an effort here. My Achilles didn't have time to properly heal. I'm struggling to walk so that's kind of a tell-tale sign that I should try to recover."
Williams added that she believes it will take up to six weeks for the injury to recover, but while that will almost certainly end her season, it should not impact her preparations for the 2021 Australian Open in January, should the Grand Slam go ahead.