Rugby Park will host Scotland's autumn Test meeting with Tonga on 22 November.
The Ayrshire ground is the home of Kilmarnock Football Club and recently had an artificial pitch installed.
Argentina will be at Murrayfield for Vern Cotter's first home match as Scotland coach on 8 November, a week before New Zealand visit Edinburgh.
Scottish Rugby Union chief executive Mark Dodson said: "We are delighted to bring international rugby to Ayrshire for the first time."
Tonga won on their last trip to Scotland, a 21-15 victory at Aberdeen FC's Pittodrie Stadium.
But Cotter won three of his four opening Tests with Scotland this summer, including a defeat of Argentina.
"It's an area where clubs, schools and local authorities are all working together to provide opportunities for our young people to play rugby," said Dodson.
"We know, from the matches we've taken to Aberdeen in the recent past, that international rugby can excite and enthral a whole new audience and as a means of boosting playing numbers it certainly helps the teachers, coaches and volunteers in all they do to encourage participation."
The 18,000-capacity Rugby Park has previously been used for Scotland football internationals and will become the nine venue in the country to host the national rugby team.
"Clearly it's home to Kilmarnock FC but it was the original home of Kilmarnock Rugby Club, so I suppose you could say that rugby is coming home," Dodson added.