| Competition: | Regionalliga |
| Season: | 2000–01 |
| Winners: | 1. FC Union Berlin, Karlsruher SC |
| Promoted: | 1. FC Union Berlin, SV Babelsberg 03, Karlsruher SC, 1. FC Schweinfurt 05 |
| Relegated: | SV Wilhelmshaven, FC Sachsen Leipzig, Lüneburger SK, Borussia Dortmund (A), Tennis Borussia Berlin, TSV 1860 München (A), SC Pfullendorf, FC Carl Zeiss Jena |
| League Topscorer: | Daniel Teixeira – 32 (KFC Uerdingen 05 14 and 1. FC Union Berlin 18) |
| Prevseason: | 1999–2000 |
| Nextseason: | 2001–02 |
The 2000–01 season of the Regionalliga was the seventh season of the league at tier three (III) of the German football league system.
The Regionalliga was split into two divisions, the Regionalliga Nord and the Regionalliga Süd. The champions of each, 1. FC Union Berlin and Karlsruher SC, were promoted to the 2001–02 2. Bundesliga, along with northern runners-up SV Babelsberg 03 and southern third placed team 1. FC Schweinfurt 05. The southern runners-up, VfB Stuttgart II, was ineligible for promotion.
| Goals | Nat. | Player | Team |
|---|---|---|---|
| 18 |  | Marko Barlecaj | SC Pfullendorf |
| 16 |  | Antonio Di Salvo | FC Bayern München (Amateure) |
| 14 |  | Danny Fuchs | TSV 1860 München (Amateure) |
| Michael Fersch | SSV Jahn Regensburg | |
| 12 |  | Berkant Göktan | FC Bayern München (Amateure) |
 | Joseph Aziz | SV Eintracht Trier 05 | |
| Michael Petry | VfR Mannheim |
