| Champ: |  Serena Williams |
| Runner: |  Sara Errani |
| Score: | 6–3, 6–0 |
| Draw: | 56 |
| Seeds: | 16 |
See main article: 2014 Italian Open (tennis).
Defending champion Serena Williams defeated Sara Errani in the final, 6–3, 6–0 to win the women's singles tennis title at the 2014 Italian Open.
Maria Sharapova's loss in the third round ended her 16-match winning streak in the tournament dating back to 2008. The streak includes two Italian Open titles. She would go on to win her third and last title in Rome in following year.
The top eight seeds receive a bye into the second round.
See also: 1 and 1. Serena Williams (champion)
See also: 2 and 4. 
Li Na (quarterfinals)
See also: 3 and 3. 
Agnieszka Radwańska (quarterfinals)
See also: 4 and 2. 
Simona Halep (third round, withdrew because of an abdominal injury)
See also: 5 and 1. 
Petra Kvitová (second round)
See also: 6 and 3. 
Jelena Janković (semifinals)
See also: 7 and 4. Angelique Kerber (second round)
See also: 8 and 2. 
Maria Sharapova (third round)
See also: 9 and 1. 
Dominika Cibulková (first round)
See also: 10 and 4. Sara Errani (final)
See also: 11 and 2. Ana Ivanovic (semifinals)
See also: 12 and 3. Flavia Pennetta (third round)
See also: 13 and 2. 
Carla Suárez Navarro (quarterfinals)
See also: 14. Caroline Wozniacki (withdrew because of a knee injury)
See also: 15 and 4. Sabine Lisicki (first round)
See also: 16 and 1. Sloane Stephens (second round)
See also: 17 and 3. 
Eugenie Bouchard (first round)Click on the seed number of a player to go to their draw section.
María Teresa Torró Flor (first round) Jana Čepelová (first round)
Casey Dellacqua (qualified) Christina McHale (qualified)
Marina Erakovic (first round) Zheng Jie (first round) Vania King (first round) Lauren Davis (qualified)
Paula Ormaechea (qualifying competition, lucky loser) Urszula Radwańska (qualifying competition)
Virginie Razzano (qualifying competition, retired) Monica Niculescu (qualifying competition) Petra Cetkovská Casey Dellacqua Christina McHale Lauren Davis