The Azzurri take on Croatia in an important Euro 2016 qualifier at San Siro on Sunday, but will now be unable to call upon the services of the Liverpool striker.
A statement released by the Italian Football Association (FIGC) on Saturday morning announced that the 24-year-old had picked up a "hamstring strain" and "mild groin" pain in training on Friday.
Balotelli - who has yet to score in the Premier League for Liverpool - received his first call-up to the national team under new coach Antonio Conte, but will now have to wait to return to the international fray.
Juventus defender Angelo Ogbonna similarly suffered a hamstring injury and will sit out the forthcoming fixtures as a precaution.
Both Italy and Croatia have 100 per cent records in qualifying so far and are joint-leaders of Group H with nine points each.
Conte's men picked up a slender 1-0 win at Malta last time out, which was preceded by victories at home to Azerbaijan and away to Norway.
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