The Wallabies will face Japan in Oita on Saturday 23 October, as confirmed by Japan Rugby Union.
The match will kick off the Spring Tour, which will see Australia head to Europe to face Scotland, England and Wales.
It will be their first meeting for the two nations since November 2017, when the Wallabies defeated the Brave Blossoms 63-30 in Yokohama.
“We’re really looking forward to squaring off against Japan. They showed everyone at the World Cup in 2019 that they are a top nation and can match it with the best," Wallabies coach Dave Rennie said in a statement.
“Jamie (Joseph) and Tony (Brown) have built a formidable squad that plays a really exciting, up-tempo style. We need to continue to test ourselves with this variety of play, so this is an important match for us as we continue to build ahead of the next World Cup in France.
“It’s been a really big year already for our squad with some starting back in February for Super Rugby AU so we will have to continue to manage the lads over the next month or two and into the Spring Tour in the UK.”
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“Today’s announcement is tremendous news that will be welcomed by Brave Blossoms fans in Japan and around the world. As a team, we’re very much aware of the significance of this match, being almost two years to the day that Japan last played international rugby at home, in our Rugby World Cup quarter final," his Japanese counterpart Jamie Joseph said.
“The opportunity to play a test match on home soil, against a team the quality of Australia, is a hugely important moment for us to reconnect with our fans and the wider Japanese public, following the success of the 2019 World Cup. “The Wallabies are a strong team, with quality players throughout their squad, like Japan, they look to play an exciting brand of fast, attacking, running rugby.
"The match will be a great opportunity for us to play our style of rugby against a side that brings something different to the test matches we have played in recent years. Both Japan and Australia will be heading onto Europe following the match, so both teams we will be looking to make the best possible start to their upcoming tours."
2021 Spring Tour (all times local)
Japan v Wallabies at Oita Stadium, Oita on Saturday 23 October at 1.45pm JST
Scotland v Wallabies at Murrayfield, Edinburgh on Sunday 7 November at 2.15pm GMT
England v Wallabies at Twickenham, London on Saturday 13 November at 5.30pm GMT
Wales v Wallabies at Principality Stadium, Cardiff on Saturday 20 November at 5.30pm GMT