Ajla Tomljanovic will play only her second grand slam tournament in almost two years after being awarded a French Open wildcard.
Adam Walton also received a wildcard into the men's main draw, boosting Australia's representation in the singles at Roland Garros to at least 11 players.
Another 13 Aussies have entered qualifying for the Paris clay-court major starting on May 26.
Tomljanovic, a former world No.32 and three-time grand slam singles quarter-finalist, made a winning return at a WTA 125 tournament in Parma, Italy, this week after being sidelined since February.
The 30-year-old is looking forward to her ninth main-draw appearance in Paris.
"I'm excited to be back playing and so grateful to have the opportunity to play Roland Garros. I'm looking forward to getting out there and doing my best," Tomljanovic said.
The world No.219 has an impressive record at Roland Garros, with her best result a fourth-round appearance in 2014.
Tomljanovic has also recorded two top-five wins in Paris, including beating fifth seed Anett Kontaveit in the opening round in her most recent appearance in 2022.
Walton, sitting at a career-high world ranking of No.111, has been recognised for his rapid improvement this season.
The 25-year-old from Brisbane has tallied 32 match wins and contested four ATP Challenger finals in 2024, boosting his ranking up more than 60 places since January.
Walton's season highlights include scoring two top-100 victories to qualify at an ATP 250 tournament in Adelaide, and making his ATP Masters 1000 main-draw debut in Miami as a qualifier.
"This will be my first time at Roland Garros and I'm incredibly thankful for the opportunity to play main draw," said Walton, a winner of 14 of his past 18 matches.
Tomljanovic joins compatriot Daria Saville in the women's singles main draw, while Walton bolsters the number of Australians in the men's event to nine.
World No.11 Alex de Minaur will lead the charge, supported by Jordan Thompson, Alexei Popyrin, Chris O'Connell, Aleksandar Vukic, Rinky Hijikata, Max Purcell and Thanasi Kokkinakis.
James Duckworth, Marc Polmans, Li Tu, Omar Jasika, Dane Sweeny, Arina Rodionova, Astra Sharma, Taylah Preston, Olivia Gadecki, Kimberly Birrell, Talia Gibson, Destanee Aiava and Priscilla Hon will all try to qualify from May 20.