The Arsenal man is suffering from an illness and has returned to his club. Chelsea striker Daniel Sturridge missed training on Monday morning due to stomach cramps but the England medical team remain hopeful he will be fit for the Group H encounter.
With his options now looking thin - Chelsea centre-half John Terry also withdrew on Sunday - England coach Roy Hodgson added three players to his squad in the form of Liverpool teenager Sterling, Tottenham's Livermore and Southampton's Lallana.
Hodgson admitted Walcott was too poorly to be considered for Tuesday night's match but is more hopeful over Sturridge's fitness.
"Theo has picked up some sort of virus," he said. "He was quite violently ill last night.
"Daniel trained yesterday [Sunday] with stomach cramps but today woke up with the same stomach cramps, and the doctor advised us to keep him out of training and see how things are. The doctor is quite confident he will be okay, though.
"I can only hope nothing is going round the squad. That was why we were quite keen to get Theo home as soon as possible and also keep Daniel away from the squad.
"I have to hope it's just an isolated case with the two players."
Sterling, Livermore and Lallana will now be given the chance to gain some valuable experience with the senior squad, according to Hodgson.
"It was a good opportunity to invite some young players to come in and be part of the atmosphere," he said.
"We still have enough good players to be able to deal with the game, but it's always good when you have the chance to invite these players to come in and see how things are."
Hodgson's only team dilemma is who will replace Terry at centre-back, with both Gary Cahill and Phil Jagielka in the running.
And the manager admits both players are more than capable of plugging the gap, although he has already made up his mind which one will run out against Ukraine.
"Both Cahill and Jagielka are good players," he said. "Whichever player I chose I'll be happy the team is well covered and that John's absence will be well covered.
"I trust them both, both are good enough. It's up to me now to choose which one I think is the better player for this match.
"I've done that but I don't want to announce anything until I've spoken to the players."