Marc Leishman has joined fellow Australian Adam Scott by withdrawing from Olympic consideration, citing health concerns for his wife who continues to recover from a near-fatal illness.
Leishman, who was in line for selection with world No.1 Jason Day after Scott declared he was unavailable, said it was a decision he did not take lightly.
"It is with deep regret that I have informed Golf Australia that I will be unable to represent my country in the upcoming 2016 Summer Olympic Games in Brazil," Leishman said.
"Many of you may know that last April my children and I almost lost my wife, Audrey, to toxic shock syndrome.
"Since then Audrey has been prone to infection and is far removed from 100 percent recovery of her immune system.
"We have consulted with Audrey's physician and due to her ongoing recovery from toxic shock and potential risks associated with the transmission of the Zika virus, it was a difficult yet easy decision not to participate."
Leishman missed the 2015 Masters to be by her side when she was given just five per cent chance of survival.
While Scott faced heavy criticism from some quarters, including golf legends Gary Player and Jack Nicklaus and Olympic legend Dawn Fraser, Leishman was hopeful people would understand his decision.
"I cannot risk placing her health in jeopardy," he continued.
"The Masters and the Olympics are the two biggest tournaments to which a golfer can be invited; however, my family will always come before golf.
"Finally, I hope that my fellow countrymen understand why I have made this decision. I hope none of them are ever in a position to have to do the same."
Golf returns to the Olympic games for the first time in 112 years with teams to be finalised on July 11 based on world rankings.
With Scott and Leishman on the sidelines, Marcus Fraser at 63rd is the new man in the hot seat.
Matt Jones (71st), Scott Hend (76th), Steven Bowditch (96th), John Senden (103rd) and Cameron Smith (109th) are the other players with a chance of making a move into the team.