The pair clashed on the hour mark in the Stadio Olimpico in a row after the Portuguese star played the ball while Edin Dzeko was down and needed treatment.
Ronaldo made a gesture with his hand with the former Real Madrid star heard saying: “You’re too small to talk.”
The Portuguese player than seemed to apologise and to hug Florenzi.
Dzeko set up Florenzi for the first goal after 79 minutes with the Bosnian then slotting in the second two minutes into injury time as Juventus fell to just their third defeat this season.
The win keeps Roma’s Champions League hopes alive with Claudio Ranieri’s side sixth, but just one point off Inter Milan who are fourth, before they play Chievo on Monday.
The defeat came on the day that Juventus displayed their new strip which no longer features the club’s iconic black and white stripes in a breakaway from over a century of tradition.
Ronaldo’s hopes of finishing the season as the Serie A top scorer are slim as his 21 goals are five behind Sampdoria’s Fabio Quagliarella’s 26 with two games left.
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