The 2000–01 Segunda División B season was the 24th edition of the tournament.
Playoffs de Ascenso: Relegated from Segunda División: Promoted from Tercera División: Relegated: Dissolved: Occupied the vacant spots by administrative relegations: Occupied the vacant spots by clubs dissolutions: Teams from Asturias, Canary Islands, Castile and León, Castilla–La Mancha, Community of Madrid and Galicia.
Teams from Aragon, Basque Country, Cantabria, Castile and León, La Rioja and Navarre.
Teams from Balearic Islands, Castilla–La Mancha, Catalonia, Region of Murcia and Valencian Community.
Teams from Andalusia, Castilla–La Mancha, Ceuta, Extremadura and Melilla.