The women performed parts of "Coat of Many Colors," "Jolene" and "9 to 5" for the star-studded audience.
Parton, who earlier in the evening won the Tex Ritter Award for the success of her autobiographical TV movie Coat of Many Colors, was in the news recently after singer Rory Feek said Parton sent his dying wife, Joey, an inspiring video message.
Joey Feek was also honored at the ACMs.
"A few weeks ago, we lost a member of our country music family with the passing of Joey Feek," presenters Martina McBride and Darius Rucker took turns reading the monitor about halfway through the show, as photos of Joey appeared on a screen and her voice could be heard singing.
"Joey was more than a wonderful singer and an ACM-winning artist, along with her husband, Rory. She was a beloved mom, daughter, family member and friend. Of all of the different ways we could honor Joey, we think that what would mean the most to Joey and her family is just one more time, for all of us -- all of her fans and all of us who love her -- to give her a big round of applause."
The crowd, which included Joey's stepdaughter Hopie and a close family friend, cheered and gave a standing ovation for the recording artist, who died of cancer last month at age 40.