Kyle Walker, Kalvin Phillips and Harry Maguire named in familiar looking England squad
Brentford striker Ivan Toney, who had two loans at Barnsley, is included despite being charged with breaching Football Association betting rules.
He is the only uncapped player as a new post-World Cup cycle begins with European Championship qualifiers against Italy and Ukraine.
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Southgate called it "a complicated situation" with “no conclusive evidence to leave him out."
Toney was overlooked for the World Cup but Southgate commented: "He is available for his club and there has not been a trial or judgement yet. I don’t know on what basis we wouldn’t pick him."
Phillips has struggled since leaving home-town club Leeds United for Manchester City. Since Qatar he has played only 55 Premier League minutes plus stoppage time.
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Hide AdFormer Sheffield United and Hull City centre-back Maguire has played 199 league minutes.
But Southgate is nothing if not loyal to those who have served him well. Ten of the 25 players selected were at the 2018 World Cup. It would have been 11 but for an injury to Raheem Sterling.
Ben White, once of Leeds, missed the cut having flown home early from the World Cup for personal reasons. Southgate argued it was better for Liverpool's Trent Alexander-Arnold to "re-energise" than be England's third-choice right-back.
Marc Guehi is preferred at centre-back to ex-Sheffield United loanee Conor Coady and former Hull City loanee Fikayo Tomori, in good form for AC Milan. Barnsley-born John Stones keeps his place.
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Hide AdThe Three Lions are in Italy on Thursday before hosting Ukraine at Wembley on Sunday.
SQUAD: Pickford, Pope, Ramsdale; Chilwell, Dier, Guehi, James, Maguire, Shaw, Stones, Trippier, Walker; Bellingham, Gallagher, Henderson, Maddison, Mount, Phillips, Rice; Foden, Grealish, Kane, Rashford, Saka, Toney.