Al Wakrah lost 2-0 to Qatar Stars League leaders Al Duhail - which left the team seven points off the pace - while Tractor lost ground on table-topper Sepahan after a 1-0 loss to Zob Ahan.
The match will be held in Group A at the Al Janoub Stadium on Wednesday.
The top two teams from each group qualify for the knockout stage.
The 2024–25 AFC Champions League Two group stage will be played from September 17 to December 5, 2024.