The 2000–01 season of the Regionalliga was the seventh season of the league at tier three (III) of the German football league system.
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.