Coach Patrice Carteron has been forced to leave out regulars Diarra, Abdou Traore from Girondins Bordeaux, Mustapha Yatabare of Guingamp and Wolverhampton Wanderers midfielder Tongo Hamed Doumbia, who were all unable to take part in the final week of preparation in Bamako.
The coach, confirming his squad as Mali departed for the tournament on Wednesday, had a change of heart over Serie A striker Mamadou Samassa, who was also injured but is now expected to recover by the time Mali meet Niger in their opening Group B game in Port Elizabeth on January 20.
Samassa was replaced in the preliminary squad by French-born Mana Dembele at the weekend but is now restored to the final, 23-man list.
Diarra's absence means the influential Fulham midfielder misses a second successive tournament, having played in four finals before that. He hurt a knee playing for Mali in the qualifiers against Botswana in September.
Last week his club manager Martin Jol warned he would not make the finals. "He can't play for us and I don't think he'll be available for them," he told the Fulham website.
Paris St Germain's Sissoko, previously on the books at Liverpool and Juventus, has not played for Mali since the 2010 Nations Cup tournament in Angola.
This season he has competed in just three matches for his French club.
Mali also play the Democratic Republic of Congo and Ghana in their opening-round group.
Goalkeepers: Soumbeyla Diakite (Stade Malien), Mamadou Samassa (Guingamp,), Aly Yirango (Djoliba AC)
Defenders: Adama Coulibaly (AJ Auxerre, France), Idrissa Coulibaly (Al Lekihwiya), Ousmane Coulibaly (Stade Brest), Salif Coulibaly (Djoliba), Fousseiny Diawara (AJ Ajaccio), Mahamadou Ndiaye (Vitoria Guimaraes), Adama Tamboura (Randers), Molla Wague (SM Caen)
Midfielders: Samba Diakite (Queens Park Rangers), Seydou Keita (Dalian Aerbin), Momo Sissoko (Paris St Germain), Samba Sow (Racing Lens), Kalilou Traore (Sochaux), Mahamane Traore (Nice), Sambou Yatabare (Bastia)
Forwards: Cheick Tidiane Diabate (Girondins Bordeaux), Cheick Fantamady Diarra (Stade Rennes), Sigamary Diarra (AC Ajaccio), Modibo Maiga (West Ham United), Mamadou Samassa (Chievo Verona). (Reporting by Mark Gleeson in Cape Town; Editing by Clare Fallon)
- Sports & Recreation