Poole Town Football Club is currently based at Tatnam Park.
Plans to turn part of a recreation ground into a football stadium have been refused.
Poole Town Football Club had applied to build a 160-seat facility at Branksome Rec in the town.
The borough council's planning committee voted unanimously against the plans, at a meeting at the Port of Poole ferry terminal.
A planning officer's report recommended councillors refuse the scheme because of the loss of open space.
Officers also felt the scheme would harm the character of the recreation ground.
Leisure councillor Peter Adams said: "As the landowner, the council was approached by Poole Town Football Club with a proposal for Branksome recreation ground.
"While the council broadly accepted the principle of the proposal, its support was subject to the football club being able to achieve a number of conditions, including planning permission.
"The application was unable to meet the criteria required to satisfy local planning policies or members of the planning committee."
A petition organised by residents, who set up Branksome Rec Action Group, collected more than 500 signatures against the plan.
Last season Poole Town won the Wessex League Premier Division, but were denied promotion to the Southern League because their current facilities at Tatnam Park, School Lane, do not meet Football Association regulations.
The club said it is already in talks with another landowner elsewhere in the borough about submitting a new application.
Chris Reeves, vice chairman of Poole Town FC, said: "The officer's report was a shock so instinctively I knew we were going to lose.
"I think our plan B will be well-received and will have cross-party support. The landowner is very keen on putting together a joint venture with Poole Town Football Club.
"This football club is not going to lay down and die."