"The Court of Federal Justice has rejected the appeal filed by Roma and has confirmed the closure of the Curva Sud for the first home match...while also confirming a fine of 50,000 euros (£42,500)," the Italian Football Federation (FIGC) said on its website (www.figc.it) on Friday.
Roma were handed the punishment after hardcore fans in that section of the Stadio Olimpico racially abused AC Milan striker Balotelli during their last game of the season against Napoli.
The Italian international has long been a target for Serie A crowds with fans of Juventus, Inter Milan and Roma repeat offenders.
Roma's match at Milan last month was stopped for two minutes after monkey chants from the away supporters were aimed at former Manchester City forward Balotelli.
Warnings were broadcast over the public address system before play restarted.
Balotelli was also targeted by Fiorentina fans last month, prompting the striker to take to Twitter to complain.