| Competition: | Segunda División |
| Season: | 2000–01 |
| Winners: | Sevilla FC |
| Promoted: | Sevilla Real Betis CD Tenerife |
| Relegated: | SD Compostela Universidad de Las Palmas CF Getafe CF UE Lleida |
| League Topscorer: |  Salva Ballesta |
| Matches: | 462 |
| Total Goals: | 1016 |
| Prevseason: | 1999–2000 |
| Nextseason: | 2001–02 |
The 2000–01 Segunda División season saw 22 teams participate in the second flight Spanish league. The teams that promoted to La Liga (top division) were Sevilla FC, Real Betis and CD Tenerife. The teams that relegated to Segunda División B were SD Compostela, Universidad de Las Palmas CF, Getafe CF and UE Lleida.
| Team | Home city | Stadium | Capacity | |
|---|---|---|---|---|
| Albacete | Albacete | Carlos Belmonte | 18,000 | |
| Atlético de Madrid | Madrid | Vicente Calderón | 54,907 | |
| Badajoz | Badajoz | Nuevo Vivero | 15,198 | |
| Real Betis | Seville | Manuel Ruíz de Lopera | 52,132 | |
| Compostela | Santiago de Compostela | San Lázaro | 12,000 | |
| Córdoba | Córdoba | Nuevo Arcángel | 21,822 | |
| Eibar | Eibar | Ipurua | 5,000 | |
| Elche | Elche | Martínez Valero | 36,017 | |
| Extremadura | Almendralejo | Francisco de la Hera | 11,580 | |
| Getafe | Getafe | Coliseum Alfonso Pérez | 17,393 | |
| Leganés | Leganés | Butarque | 8,138 | |
| Levante | Valencia | Ciutat de València | 26,354 | |
| Lleida | Lleida | Camp d'Esports | 13,500 | |
| Real Jaén | Jaén | La Victoria | 11,500 | |
| Real Murcia | Murcia | La Condomina | 16,000 | |
| Racing Ferrol | Ferrol | A Malata | 12,024 | |
| Recreativo de Huelva | Huelva | Colombino | 13,000 | |
| Salamanca | Villares de la Reina | Helmántico | 17,341 | |
| Sevilla | Seville | Ramón Sánchez Pizjuán | 45,500 | |
| Sporting de Gijón | Gijón | El Molinón | 25,885 | |
| Tenerife | Santa Cruz de Tenerife | Heliodoro Rodríguez López | 20,000 | |
| Universidad de Las Palmas | Las Palmas | Municipal de Maspalomas | 6,000 |
| Autonomous community | Number of teams | Teams | |
|---|---|---|---|
| 1 | 5 | Betis, Córdoba, Real Jaén, Recreativo, Sevilla | |
| 2 | 3 | Atlético Madrid, Getafe, Leganés | |
| 3 | 2 | Tenerife, Universidad de Las Palmas | |
| 2 | Badajoz, Extremadura | ||
| 2 | Compostela, Racing de Ferrol | ||
| 2 | Elche, Levante | ||
| 7 | 1 | Sporting | |
| 1 | Eibar | ||
| 1 | Albacete | ||
| 1 | Salamanca | ||
| 1 | Lleida | ||
| 1 | Murcia | ||