| Champ: |  Serena Williams |
| Runner: | Madison Keys |
| Score: | 7–6(7–5), 6–3 |
| Draw: | 56 (8 / 3) |
| Seeds: | 16 |
See main article: 2016 Italian Open (tennis).
Serena Williams defeated Madison Keys in the final, 7–6(7–5), 6–3 to win the women's singles tennis title at the 2016 Italian Open. This was Williams's second title in Rome won without dropping a set, and her fourth Italian Open title overall.
Maria Sharapova was the reigning champion, but could not defend her title because of her provisional suspension after failing a drug test at the Australian Open.[1]
The top eight seeds received a bye into the second round.