The 23-year-old Montenegrin international has been a long-term target for the Premier League champions, who tried to bring Jovetic to England during the summer.
Citizens boss Roberto Mancini failed to land a string of transfer priorities before the start of the season, including Javi Martinez and Robin van Persie - who opted instead to join local rivals Manchester United.
City were planning to renew interest in Jovetic in January, but their hopes of luring him to the Etihad Stadium in the New Year have been dealt a significant blow, after he revealed his intentions to remain in Italy.
"Leave Florence? This is not what I am thinking about," said the Serie A star. "I'd rather stay in Italy because I know the language and my family is happy here."
Manchester City have slipped six points behind Manchester United in the Premier League title race, and having been humiliated in the Champions League after being knocked out at the group stage for the second consecutive season, Mancini will ask to be backed when the window reopens next month.
However, he will likely have to turn his attention to alternative targets, with Jovetic committing his future to the Fiorentina cause.