| Competition: | Serie B TIM |
| Season: | 2000–01 |
| Promoted: | Torino (3rd title) Piacenza Chievo Venezia |
| Relegated: | Pescara Ravenna Monza Treviso |
| League Topscorer: | Nicola Caccia (23 goals) |
| Matches: | 380 |
| Total Goals: | 937 |
| Prevseason: | 1999–2000 |
| Nextseason: | 2001–02 |
The Serie B 2000–01 was the sixty-ninth tournament of this competition played in Italy since its creation.
Siena, Crotone, Cittadella and Ancona had been promoted from Serie C, while Torino, Venezia FC, Cagliari and Piacenza had been relegated from Serie A.
| Team | Manager | Kit manufacturer | Shirt sponsor | |
|---|---|---|---|---|
| Ancona |  Fabio Brini | Ropam | Carilo/Banca Marche | |
| Cagliari | Giuseppe Materazzi | Uhlsport | Pecorino Sardo/Terra Sarda | |
| Chievo Verona | Luigi Delneri | Hummel | Paluani | |
| Cittadella | Ezio Glerean | Hummel | Imasaf | |
| Cosenza | Bortolo Mutti | Legea | Provincia di Cosenza | |
| Crotone | Giuseppe Papadopulo | Devis | Caffè Guglielmo | |
| Empoli | Silvio Baldini | Erreà | Computer Gross | |
| Genoa | Franco Scoglio | Kappa | None | |
| Monza | Gaetano Salvemini | Legea | ghostyclub.com | |
| Pescara | Delio Rossi | Puma | Gelati Gis | |
| Piacenza | Walter Novellino | Lotto | Copra Ristorazione Servizi (H)/Gruppo DAC Arco (A) | |
| Pistoiese | Walter Nicoletti | Erreà | Vannucci Piante | |
| Ravenna | Stefano Di Chiara | Erreà | Associazione italiana contro le leucemie-linfomi e mieloma | |
| Salernitana | Francesco Oddo | In-house | ICS Personal Computer/Compuprint/Exigo Jeans | |
| Sampdoria | Luigi Cagni | Asics | Dreamcast | |
| Siena | Antonio Sala | Lotto | Romagest | |
| Ternana | Andrea Agostinelli | Erreà | ONBanca | |
| Torino | Giancarlo Camolese | Kelme | Directa SIM | |
| Treviso | Mauro Sandreani | Lotto | Segafredo | |
| Venezia | Cesare Prandelli | Kronos | Emmezeta |