The world's most famous painting should be returned to the Uffizi museum where was displayed early in the 20th Century, according to the National Committee for Historical, Cultural and Environmental Heritage, which organised the petition, the 'Daily Mail' reported.
Committee President Silvano Vincenti said he has made a formal request to the French minister of culture, Aurelie Filippetti, for the painting to be given back.
He said the return of the painting would be of 'high historical value, both 'symbolic' and 'moral'.
But the Louvre museum itself has already snubbed the committee. And Florence's claims on the Renaissance masterpiece, known by Italians as La Gioconda, might not be that straightforward.
Leonardo is thought to have begun work on the enigmatic portrait of Lisa del Giocondo, the wife of a wealthy Tuscan silk merchant, in Florence in 1503. But art historians think he took it with him when he moved to France in 1516.
The French Royal family then acquired it and following a spell at Versailles it ended up at the Louvre museum after the French Revolution.
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It was stolen from the Paris museum in 1911 and was discovered two years later at the Florence home of the Italian patriot and former Louvre employee Vincenzo Peruggia.
However, the painting was exhibited only briefly in the Uffizi and then in Rome before it was returned to the Louvre that year.
Mona Lisa is a 16th century portrait painted in oil on a poplar panel by da Vinci during the Italian Renaissance.
The work is owned by the French government and hangs in the Louvre museum in Paris.
The painting is generally acknowledged to be the most famous work of art in the world and debate has raged for years over the reason for her famously enigmatic smile.