2000–01 Cypriot Fourth Division

The 2000–01 Cypriot Fourth Division was the 16th season of the Cypriot fourth-level football league. Sourouklis Troullon won their 1st title.

Format
Fourteen teams participated in the 2000–01 Cypriot Fourth Division. All teams played against each other twice, once at their home and once away. The team with the most points at the end of the season crowned champions. The first three teams were promoted to the 2001–02 Cypriot Third Division and the last three teams were relegated to regional leagues.

Point system
Teams received three points for a win, one point for a draw and zero points for a loss.

Changes from previous season
Teams promoted to 2000–01 Cypriot Third Division
 * MEAP Nisou
 * Elia Lythrodonta
 * THOI Lakatamia
 * AMEP Parekklisia

Teams relegated from 1999–2000 Cypriot Third Division
 * Achyronas Liopetriou
 * Ellinismos Akakiou
 * Doxa Paliometochou

Teams promoted from regional leagues
 * Anagennisi Lythrodonta
 * Anagennisi Prosfigon Lemesou
 * ASPIS Pylas
 * Sourouklis Troullon

Teams relegated to regional leagues
 * AEK Kythreas
 * Orfeas Nicosia
 * PAOK Kalou Choriou