The 31-year-old forward, who has lost his place in Manchester United's first team, was also left out by England earlier this year for matches against Germany and Lithuania.
Southgate named a 25-man squad which includes Tottenham Hotspur right back Kieran Trippier for the first time.
Sunderland striker Jermain Defoe, who scored on his return to the international scene against Lithuania in March after a long absence, retains his place in Southgate's plans.
