The Old Lady have named former Italy manager the veteran manager as their manager until the end of the season.
The experienced coach takes over from his predecessor, who was dismissed after the Serie A side's defeat by Lazio on Sunday.
Tudor arrived at the club in March as Thiago Motta's replacement but departed after seven months after an eight-game winless streak.
Spalletti led Napoli to the Serie A title in 2022-23 prior to assuming the Italy role, which missed out on the 2022 World Cup during Roberto Mancini's tenure.
He helped them qualify for Euro 2024 exiting in the last 16 stage against Switzerland.
Yet, the Italian FA let Spalletti go in June after a World Cup qualifying defeat by Norway, as Italy faced the possibility of not making a third World Cup in a row.
He has previously coached Inter Milan, Roma, and Udinese in Italy's top flight and secured two Russian Premier League titles with Zenit St Petersburg.
Juventus were temporarily led by Massimo Brambilla in the midweek 3-1 triumph over Udinese moving them to seventh position in the league - six points adrift of Napoli at the top after nine rounds.
Their Champions League campaign sees them in 25th place in the group stage, having lost once and drawn twice in their opening three matches.