Siena and Padova have officially gone bankrupt after failing to register for the Serie B and Lega Pro seasons.
In Italy each club must prove its financial stability before beginning a new campaign and failure to do so results in not being registered for the season.
Siena had been on the verge of collapse for several years, as their main backers the Banca Montepaschi di Siena were caught up in a financial scandal.
President Massimo Mezzaroma resigned earlier this month in order to help find a new buyer quickly, but Siena could not be saved.
They missed today’s deadline to provide paperwork covering taxes and unpaid wages, as their debts amounted to €70m (£56m).
Siena had been founded in 1904 and were in Serie A most recently in 2012-13.
The club title can be bought by a new owner, but they’ll have to begin again from Serie D.
Padova failed to register for the Lega Pro campaign after their relegation, so will be able to resume under a new owner from the amateur Leagues.
Padova Calcio were founded in 1910 and were most recently in Serie A in 1995-96.
Grosseto were also at risk, but were able to prove their solidity before the deadline.
It now remains to be seen whether a club will be plucked from Lega Pro to fill in for Siena or the division reduced.