The greenbelt development was being promoted by Judy Murray in the face of strong local opposition.
Planning officers at the council had earlier recommended that the Park of Keir application be refused.
Councillors said they had decided to reject the bid after carefully considering the arguments for and against the project.
The proposed development, between Dunblane and Bridge of Allan, would have included tennis and golf facilities along with a visitor centre and museum, all set in a new country park.
The plans also included luxury homes to help pay for the scheme.
But planning officers said the application should be refused because it was sited on greenbelt land.
They also said there was not enough affordable housing proposed and added the residential element was contrary to Scottish planning policy, because residents would have to travel for basic amenities and services.
The scheme had received high-profile support from former Manchester United manager Sir Alex Ferguson and golfer Colin Montgomerie.
However, campaigners said the loss of greenbelt land to the development was too high a price to pay. There were more than 1,000 objections to the scheme and only 45 in support.
Mark Ruskell, Scottish Green Party councillor for Dunblane and Bridge of Allan, spoke against the proposal at Tuesday morning's hearing on the plans.
Following the decision, Mr Ruskell said: "I'm delighted that this proposal has been rejected. It would have been totally wrong to rip up our democratically-agreed local plan to make room for executive housing on this treasured greenbelt area.
"For over 25 years, local communities have fought against development at Park of Keir. Judy Murray had a good idea but it was in completely the wrong location - I'm sure that sporting legacy can be secured elsewhere."
Ms Murray said on Friday that she wanted to leave a legacy to the success of her two sons, tennis players Andy and Jamie Murray.
A spokeswoman for Stirling Council said: "Having carefully considered and discussed extensively all the arguments for and against this proposal, Stirling Council's Planning and Regulation Panel has today refused the application for a development at Park of Keir."

Currently, they can start school at the beginning of the term in which they have their fourth birthday.
But Powys council's cabinet approved the plans on Tuesday, which will see children start school in the September.
The change will be introduced from September 2017 and will save £1.2m a year.
The council also voted to increase the hours of free pre-school provision from 10 hours per week to 12.5 hours.
Councillor Arwel Jones, cabinet member for schools, said: "There's no secret that we are proposing this revised policy to help in our bid to meet the £27m budget savings target over the next three financial years."
He added: "Today's decision will bring us in line with the majority of other councils in England and Wales where children start school in the September after their fourth birthday."
Councillors in Stirling have rejected plans for a controversial sports and housing development near Dunblane.
[NEXT_CONCEPT]
Children in Powys will only be able to start primary school after turning four years old, it has been decided.