Companies that provide parking spaces for employees will be forced to pay thousands of pounds a year if a pilot scheme gets the go-ahead.
Nottingham City Council wants to charge companies an annual fee of £185 for each of their staff car parking spaces.
If the authority gets the green light that charge will rise to £350 a year by 2014 - and business leaders fear it could then be rolled out by other councils across the country.
John Dowson, head of policy and representation at Derbyshire and Nottinghamshire Chamber of Commerce, said: "The city council is looking at being a trailblazer and showing this as the way forward for transport in the future.
"We are concerned that if they are allowed to get away with it then other local authorities will go ahead because legally they can."
The council says the levy will raise millions of pounds needed to fund the expansion of the city's tram network.
Firms with 10 spaces or fewer will be exempt, and employers can pass the charge on to their workforce. But research has shown the economic impact on the city could be severe.
So far 61 companies are threatening to relocate, while a further 74 are considering partial relocation or are shelving plans to expand in Nottingham.
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