First-seeded Margaret Smith was the four-time defending champion, and successfully defended her title, defeating Lesley Turner 6–3, 6–2 in the final to win the women's singles tennis title at the 1964 Australian Championships.
The seeded players are listed below. Margaret Smith is the champion; others show the round in which they were eliminated.
Margaret Smith(champion) Lesley Turner (finalist) Jan Lehane (semifinals) Robyn Ebbern (semifinals) Judy Tegart (quarterfinals) Madonna Schacht (quarterfinals) Jill Blackman (quarterfinals)
Rita Bentley (first round)