Mark and Kim Liddiard, who have lived on Strathallan Estate for eight years, said they have not been consulted about the new venue and only found out about the move through media reports.
T in the Park – which attracts 85,000 revellers each year – is moving to the 1,000-acre estate in Perthshire after safety fears were raised about a pipeline under the former site at Balado.
The couple, along with other residents in the 30 homes on the estate, believe the move could have a “significant effect” on local wildlife.
The area is home to bats, nesting ospreys and kites; and the Machany Water, which runs through the estate, is an important site for wild salmon spawning.
Mrs Liddiard, 56, said yesterday: “It was a real shock to us – we were really concerned as to what was going on. Nobody had a conversation with us.
“There’s real concern the Machany Water is going to get severely damaged. The reason some people come to the area is for wild salmon fishing.
“We’ve got nesting ospreys on the estate. They would also be disturbed by a concert.”
David Summers from the Tay District Salmon Fisheries Board, said: “The Machany Water is indeed a salmon spawning stream.
“We would like to know what things are planned around the stream and what precautions would be taken to ensure there is no pollution.”
Festival organisers DF Concerts has not been required by Perth & Kinross Council to lodge a planning application for the annual event.
Any landowner can temporarily change the use of their land if it is for less than 28 days in a year without needing planning permission, unless it has a “significant effect on the local environment”.
The couple are also concerned about the roads leading to the estate. Strathallan is three miles from the A9 but the main routes to the estate have single-track roads and listed small bridges, which they do not think could take the level or weight of traffic likely to go to the festival.
The Liddiards have now written to Scottish ministers asking them to review the situation and are prepared to take the matter to the Court of Session for a judicial review.
Sandy Telfer, head of planning at law firm DLA Piper Scotland, said: “The last thing that our clients want to do is to derail T in the Park.
“They and the significant number of other local residents who are concerned about developments at Strathallan are only too aware of how iconic an event it has become in Scotland.
“The problem is that they have no idea how the festival will impact on their personal and professional lives during the six to eight weeks in the summer of each year going forward that it takes to set the site up and take it down at the end of the three days of live music.”
A spokeswoman for Perth and Kinross Council said: “An environment impact assessment screening has been conducted at the identified site. This concluded that any environmental impact from the event was unlikely to be significant.”
A spokeswoman from T in the Park said: “Building a positive relationship with the local community in Strathallan is extremely important to us.
“In the short timescale we’ve had, we’ve written to, spoken to and met with as many of the residents closest to the new site as we can, and this process is ongoing.
“The feedback we’ve had so far from the people we’ve spoken to has been extremely positive and this has established excellent foundations for going forward.”