The 2000–01 National First Division was the 5th season of the second tier of South African soccer since the reorganisation in 1996.
It consisted of two streams, an Inland stream of 16 teams, and a Coastal stream of 14 teams.
The Coastal stream was won by AmaZulu and the Inland stream by Black Leopards, with both earning promotion to the 2001–02 Premier Soccer League.
[1][2] Arcadia Shepherds were relegated just short of 100 years after their formation, while Real Rovers were relegated after earning just two wins and six points all season, with a goal difference of minus 68.
Sides received R50 000 each for winning the stream, with the winner of the championship playoff receiving a further R25 000.