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Justin Timberlake, left, and Chris Stapleton perform at the 49th annual CMA Awards at the Bridgestone Arena on Wednesday, Nov. 4, 2015, in Nashville, Tenn.Chris Pizzello

Justin Timberlake, who grew up in Memphis, Tennessee, tapped into his country roots with a show-stopping performance at the Country Music Association Awards.

Timberlake sang alongside popular songwriter and singer Chris Stapleton at the Bridgestone Arena in Nashville, Tennessee, on Wednesday night. They strummed their guitars while singing "Tennessee Whiskey," which Stapleton wrote, and followed that with Timberlake's tune, "Drink You Away."

The audience was in awe: Timberlake's wife, actress Jessica Biel, watched excitedly and bopped her head, while Keith Urban recorded the performance with his phone.

Stapleton went on to win new artist and album of the year for "Traveller."

"This is unbelievable. I don't know what to say. I don't know what to think," said Stapleton who has written for Kenny Chesney, George Strait and others. "Thank you so much for the love."

Carrie Underwood and Brad Paisley were also show-stoppers with their opening jokes – including about Blake Shelton and Miranda Lambert's divorce.

"And of course we can't ignore the breakup that rocked our world," Underwood said, as the camera panned to Miss Piggy and Kermit the Frog.

"On a completely unrelated topic, howdy Blake?" Paisley replied. The camera panned to Shelton who laughed and clapped his hands, along with the audience.

"Did you hear that duet that Blake did with Ashley Madison?" Paisley asked, earning laughs from the audience at the Bridgestone Arena in Nashville, Tennessee.

"That was with Ashley Monroe," Underwood replied.

Shelton confirmed Wednesday just hours before the awards show that he and his fellow "Voice" mentor Gwen Stefani are dating. He and Lambert announced their divorce in July.

Lambert, who is also at the CMAs, is nominated for entertainer of the year, while Shelton will perform later in the show.

Duets kicked off the awards show: Eric Church and Hank Williams Jr performed "Are You Ready for the Country" and John Mellencamp and Keith Urban strummed their guitars during "Pink Houses." Sam Hunt won over the audience with his performance of "Take Your Time," while Underwood sang "Smoke Break."

Little Big Town, who also earned applause for their performance of "Girl Crush," won the first award of the live telecast for single of the year for their No. 1 hit. The writers of "Girl Crush" won song of the year.

"This is about being brave and being bold," Karen Fairchild said onstage with her group. "Country radio, thanks for having the guts for playing it."

The foursome, along with Church, walked into the arena as the most nominated act with five. They lost music video of the year to Maddie & Tae's "Girl in a Country Song" and musical event of the year to Church and Urban. The group, which includes Kimberly Schlapman, Jimi Westbrook and Phillip Sweet, is also nominated for vocal group of the year, which it has won for the last three years.

Little Big Town is not up for the biggest honour: entertainer of the year. Church, who announced he released a surprise album called "Mr. Misunderstood" on Wednesday, is nominated for the top prize, pitting him against Garth Brooks, Lambert, Chesney and last year's winner, Luke Bryan.

Kacey Musgraves, Jason Aldean, Reba McEntire and Dierks Bentley will also perform during the three-hour show, airing live on ABC.

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