The visitors lost Douglas Costa and Miralem Pjanic to suspected hamstring and thigh injuries respectively before half-time and never really got going at Stadio Artemio Franchi.
Fiorentina were without a point heading into this match - and without a Serie A win since February - but looked the more likely to score against a lacklustre Juventus, who move one point clear of Inter and Torino at the summit after playing a game more.
Franck Ribery was handed his full debut for the home side and fired in a low drive 16 minutes in that was saved by Wojciech Szczesny, who survived an earlier scare when his clearance ricocheted off Federico Chiesa and flew over the bar.
Fiorentina keeper Bartlomiej Dragowski kept out efforts from Blaise Matuidi and Pjanic in the space of a minute, but Szczesny was also kept busy at the other end as Dalbert came close with his back-post header.
The hosts continued to look the more likely to score in the early stages of the second half as Dalbert's angled drive was palmed away by Szczesny, moments before Gaetano Castrovilli scuffed wide from a good position.
Chiesa's deflected effort skimmed the crossbar 65 minutes in and Nikola Milenkovic then headed straight at Szczesny from a good position as the match finished scoreless.
What does it mean? Juve far from full flow
Juve's 4-3 win against Napoli followed a slender 1-0 victory against Parma on the opening weekend.
Kalidou Koulibaly's own goal may have rescued the champions last time out but there was no such luck today, giving the chasing pack a chance to capitalise on this first slip-up of the campaign.
Ribery still has it
Used as a substitute in Fiorentina's first two games, the two-week break gave Ribery a chance to reach peak fitness and he was very lively on his first start.
The 36-year-old had a couple of efforts on target in the first half, created a glorious chance for Dalbert and left the field to a standing ovation 69 minutes in.
De Ligt needs time to adapt
Matthijs de Ligt struggled on his Serie A debut against Napoli two weeks ago and he again looked shaky at times here as Fiorentina took the game to their opponents - though Juve did at least keep a clean sheet on this occasion.
The Netherlands centre-back put Szczesny under pressure with a poor back-pass in the first half and was cautioned 35 seconds into the second period for bringing down Ribery.
Key Opta Stats
- Juventus are winless in their last three games away from home in Serie A following an International break (D3 L1).
- Prior to today the last time Juventus made two substitutions in a first half in Serie A was in January 2011, against Parma.
- Fiorentina have not won in their last 13 home Serie A games, their longest run in the top flight.
- Juventus have won only one of their last six Serie A away games, after five wins in the previous six.
- Juventus conceded 18 shots: the last time the Bianconeri conceded more in Serie A was in March against Napoli (21).
- Fiorentina have had 18 shots in all three Serie A games this season.