Gareth Barry was ruled out of the European Championship on Monday because of a stomach injury, robbing England of one of its two holding midfielders less than two weeks before the start of the tournament.
Barry underwent a scan on the problem he picked up in the 1-0 friendly win over Norway on Saturday and results showed he has torn a muscle in his lower abdomen, the Football Association said.
The Manchester City player was replaced in the 23-man squad by Everton center back Phil Jagielka, meaning versatile defender Phil Jones is likely to act as cover for England in central midfield.
England manager Roy Hodgson also had Liverpool midfielder Jordan Henderson on the standby list but preferred Jagielka, who started the Norway match alongside Joleon Lescott at center back.
"I'm very disappointed to lose Gareth ... he has featured prominently in the England team over the last few years," Hodgson said.
"I know how much he wanted to be part of the tournament but I'm sure he will still have a part to play with England after the Euros."
Barry was one of City's best players at the end of the Premier League season as the club clinched its first title in 44 years.
Without him, Hodgson is likely to start Scott Parker alongside captain Steven Gerrard in central midfield. Frank Lampard and James Milner are the other players in the squad who can play central midfield, while Jones has played there on a number of occasions for Manchester United and also for England against Sweden in a friendly this year.
England's first match at the Euros is against France on June 11. Sweden and co-host Ukraine are also in the Group D.