Azzurri team doctor Enrico Castellacci confirmed Motta had suffered a hamstring strain and, although he was able to train, it was not certain he would be fit to start at the Olympic Stadium in Kiev.
Castellacci said: "Motta is training with other players and we will assess
Brazil-born Motta started all three of the Azzurri's group games as their most advanced midfielder.
Italy have already lost their most experienced defender, Giorgio Chiellini, to injury for the England match.
He is battling to be available should Italy reach the Euro 2012 semi-finals.
Mario Balotelli was expected to be fit to face England despite suffering a slight back problem in training.