Wharton exited the field of play after an hour of Sunday's 5-2 win over West Ham United, but Glasner confirmed the midfielder was fit at his pre-match press conference.
Holding returned to the first-team squad for the first time in 2024 in that game, but will be out of contention at Selhurst Park against Eddie Howe’s side.
“Unfortunately Rob Holding is injured again,” the manager confirmed at his pre-match press conference.
“It is tough because he is just back from injury. We don’t know the degree but it is something with his hamstring, so we don’t know how long it will take.
“He will definitely miss tomorrow's game and I think Saturday against Fulham. That is the bad news, but the other players are fit.”
Glasner says the rest of the squad will rally around Holding after what has been a frustrating season.
“[Frustrating] for all of us, especially him,” the manager said of the situation. “When you come back after a long injury and work so hard, and then see him smiling in training with the team and his first time in the squad…
“But sometimes it happens, especially if you are not used to team training or your body isn’t used to it.
“It happened, and we have to accept and learn from it, so it doesn’t happen again.”