Jaguares and Sunwolves will make their Super Rugby debuts in 2016 as Highlanders seek to defend their title in the competition's new format.
As the competition expands to 18 teams from five countries, the South African Conference splits in two to accomodate Argentine franchise Jaguares, Sunwolves of Japan and the returning Southern Kings.
The finals series has also increased from six to eight teams, with the winners of each of the four conferences guaranteed a seeded berth.
The Australian and New Zealand conferences share three further spots, with the final place going to the best runner-up across the two South African pools.
Amid the changes, last year's champions Highlanders will be aiming to claim back-to-back titles.
The only notable losses from last year's squad are back-rowers Nasi Manu and John Hardie, but the Highlanders are boosted by the addition of Luke Whitelock and Liam Squire.
However, the absence of utility back Richard Buckman is a blow for the reigning champions, after he was ruled out for the season following a shoulder injury sustained in a pre-season match against French side Racing 92.
"It is obviously pretty disappointing, I was looking forward to another big season trying to defend that title," he said.
"But it is just one of those things you can't do too much about now and look forward to the future and see the positives and get all my injuries right."
Highlanders start the campaign with an away clash against the Blues, while the other game on Friday sees the Brumbies host last year's runners-up the Hurricanes, who have lost World Cup winners Ma'a Nonu and Conrad Smith since that run to the final.
Crusaders failed to make the finals for the first time since 2001 last season - they must now pick themselves up without the help of the retired Richie McCaw and Dan Carter, who has joined Racing 92.
The new format means the South African sides have less travel throughout the season, and the Stormers will be hoping they can build on their finals appearance last year.
As for the Australian sides, the Brumbies signed experienced Argentina scrum-half Tomas Cubelli to replace Montpellier recruit Nic White, and they will be pushing to go one better this season after their semi-final loss to the eventual runners-up in 2015.
They will face some fierce competition from fellow semi-finalists the NSW Waratahs, however, with Daryl Gibson looking to impress as new coach after taking over from Wallabies boss Michael Cheika.