Almost 1,800 people have signed a petition against plans to build a mosque close to the Leaning Tower of Pisa, meaning a vote will have to take place on the project.
Egyptian-born politician, journalist and Christian convert Magdi Allam, who set up the petition called the number of signatures "the Italian revolution".
Opponents of the planned mosque claim it is too close to the Tuscan city's famed monument and tourist attraction, is a potential hotbed of radical Islam, and say its finances are murky.
"Pisa's city council has sold a plot of land to the Union of Italian Islamic Communities [Ucoii - Italy's largest Muslim grouping] which ideologically is close to the radical Muslim Brotherhood Movement," said Allam.
"The funding is from a foundation in Qatar, which has given Ucoii 25 million euros to build 33 mosques."
The referendum's supporters claim the majority of Pisa's citizens are against the planned mosque.
"A survey showed that 57 percent of are Pisans are opposed," said Gianluca Gambini, vice-chairmain of the committee set up to fight plans to be build the mosque.
"We won't stop here and are counting on getting at least 2,000 signatures, putting us in a strong position with the city council."
Less than a week ago, Italian authorities expelled a Tunisian supporter of the Islamic State jihadist group suspected of plotting an attack on the Leaning Tower of Pisa, which receives over six million visitors a year.
However Pisa's centre-left mayor Marco Filippeschi has slammed moves to block the planned mosque, saying they are discriminatory and against freedom of worship.
"It would be inadmissible discrimination to ask citizens to vote on building a mosque, so the question will be put as urban planning issue," he said.
"It's a mess intended to get around articles 3, 8 and 19 of Italy's constitution, which uphold religious freedom. And it contravenes international anti-discrimination norms," Filippeschi added.
Pisans have never complained about the small mosque where for the past 20 years the city's Muslims have prayed, he said.