The 25-year-old has withdrawn with a thigh problem, meaning he will not feature in England's match in Moldova on Friday, or Tuesday's home game against Ukraine. The injury to the former Manchester City winger was confirmed after a scan, and he has now returned to Sunderland for treatment.
No replacement has been called up for Johnson, meaning Roy Hodgson's options on the right wing will now be limited to one of Arsenal duo Theo Walcott or Alex Oxlade-Chamberlain. James Milner is expected to start on the left wing.
Johnson becomes one of several England player to pick up an injury for the games, with Wayne Rooney injured, West Ham striker Andy Carroll already withdrawn after pulling his hamstring, and Ashley Cole also missing - although the Chelsea left-back could be available for the second game against Ukraine.