Villa captain Grealish was a surprise absence from Gareth Southgate’s initial squad for the Nations League matches after his impressive form helped his club avoid relegation from the Premier League last season.
But the withdrawals of Manchester United forward Rashford and Tottenham midfielder Winks created a vacancy the 24-year-old will fill.
Grealish, recently linked with a move to Manchester United, has previously represented England at Under-21 level and was a member of Southgate’s Toulon youth tournament-winning squad in 2016.
Southgate had already added Wolves defender Conor Coady and Arsenal midfielder Ainsley Maitland-Niles to his squad on Saturday after dropping Manchester United captain Harry Maguire following the guilty verdict over in his Greek court case.
Maguire, who had pleaded not guilty, is appealing after being handed a suspended prison sentence after the street brawl.
Rashford has been nursing an ankle problem suffered towards the end of last season and he tweeted: “Gutted. Never want to let this team down but these fixtures just came a little too soon for me.
“I tried my best but I have to focus on starting the season at my strongest for club and country. Good luck boys, I’ll be cheering you on from home.”
England’s players met up at their St George’s Park training camp on Monday ahead of the clash with Iceland in Reykjavik on September 5 and the trip to face Denmark in Copenhagen three days later.