Mary Pierce defeated Conchita Martínez in the final, 6–2, 7–5 to win the women's singles tennis title at the 2000 French Open.

It was her second and last major singles title.

Pierce became the first Frenchwoman to win the title since Françoise Dürr in 1967, and remains the most recent Frenchwoman to win the tournament.

Steffi Graf was the reigning champion, but she retired from professional tennis in August 1999.

This marked the French Open main-draw debut of future champion Anastasia Myskina; she lost to Cara Black in the first round.