Manchester United's Wayne Rooney has withdrawn from the England squad with a knee injury, the Football Association announced on Monday.
Rooney will return to his club and won't be available for England's friendly against Spain of Tuesday after missing Three Lions training on Monday morning with a "minor" issue.
Ryan Bertrand, who also missed training, will remain with the national team, said the FA, who did not immediately name any replacement for Rooney.
Interim manager Gareth Southgate had said earlier on Monday that both players would be assessed to see if they will be available for selection.
After missing today's training session with a knee problem, @WayneRooney has withdrawn from the #ThreeLions squad: https://t.co/tWGx6xW3a1
- England (@England) November 14, 2016
"They weren't ready to train today so they must be a doubt, but we'll have to look later," Southgate said. "I wouldn't start Wayne in this game anyway, so we'll decide which route to go.''
Jordan Henderson -- who has stood in as skipper before -- will wear the captain's armband in place of Rooney.
If Rooney is not fit before the weekend, that would leave Manchester United manager Jose Mourinho without his top two strikers, as Zlatan Ibrahimovic is suspended.
Mourinho could be forced to start either Anthony Martial or Marcus Rashford as the central striker against Arsenal on Saturday.
While Southgate's agreed tenure expires at full-time on Tuesday, the odds are shortening on him staying on.
The team's comfortable standing in World Cup qualifying, allied to the feelgood factor of Friday's 3-0 win over old rivals Scotland, have given him a firm claim to the job.
He steadfastly refuses to discuss his own candidacy but admits that his mindset since picking up the baton from Sam Allardyce has been to plan for the future.
Rather than shore up his position with safe decisions he has shown flashes of courage -- not least in his treatment of Rooney but also in the progressive style he attempted to impose on the British derby.
And that will continue against a Spain side who appear to outgun their hosts for sheer quality.
"We've got to make the correct decisions, not the convenient decisions," he said. "To make what we believe are the right decisions for the right reasons.
"It would have been easy to say [of Scotland], 'OK, we have to win this match... we'll go solid, soak up pressure, hit teams on the break and play all our experienced players.'
"Or, do we do what we did... try to build from the back, play with some risks, involve some younger players who we think are going to be the future, work longer term.
"My view of the game is you manage every game like you're going to be there forever and make decisions for the long term.
"Tomorrow what's the plan? Go selfish, shut up shop, try to eke out a 1-0 or do we say 'no lads, let's play with belief, go with what we think is the right way to play.'
"We'll get some of those things wrong but we'll get a lot right. We have to think further forward."
Southgate said he would understand if those at the Football Association who will determine his future looked toward other options before decidiing.
"I think it's right to for everyone to reflect," he added. "If you're appointing a manager at any football club in my opinion you should take time to see what fits with your philosophy of what you want to do.
"You should speak to all the people you want to speak to because who knows, someone might emerge who you don't know so much about."