The 1964–65 Southern Football League season was the 62nd in the history of the league, an English football competition.
Weymouth won the championship, whilst Corby Town, Hereford United, Poole Town and Wimbledon were all promoted to the Premier Division.
Nine Southern League clubs applied to join the Football League at the end of the season, but none were successful.
Division One consisted of 22 clubs, including 17 clubs from the previous season and five new clubs: Also, Yiewsley changed name to Hillingdon Borough.
Alongside the four League clubs facing re-election, a total of 14 non-League clubs applied for election, including nine Southern League clubs.