Five Star Movement (M5S) leader Luigi Di Maio and Matteo Salvini, head of the nationalist League party, will meet Italian President Sergio Mattarella for consultations at the presidential palace on Thursday afternoon.
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The talks began in the morning in Rome with the smallest groups in the Italian parliament.
The M5S is Italy's largest single party after picking up just under 33 percent of the vote in the March 4 election.
Salvini's right-wing coalition, which includes Berlusconi's Forza Italia party, is the largest group with 37 percent.
Both Di Maio and Salvini have said that they are ready to govern with one another but Di Maio is demanding that Salvini break his alliance with media magnate Berlusconi, something the 45-year-old League leader has so far refused to do.
Salvini will present a united right-wing front on Thursday, going to meet Mattarella with Berlusconi and his other coalition partner Giorgia Meloni -- leader of the far-right Brothers of Italy party -- after the three went separately to the first round of talks last week.
Impasse
Salvini needs to keep his coalition intact for regional elections on April 29 in northeastern Friuli-Venezia Giulia where the League is aiming for victory. In order to win, it has to maintain its alliance with Forza Italia.
On Wednesday Alessandro Di Battista, an important figure within the M5S, said Berlusconi represented everything that was wrong with Italy.
"They are the words of the many people who believe that Berlusconi belongs to the political past," Di Maio said on current affairs programme Porta a Porta.
However an agreement between Di Maio and Salvini is currently the best hope for a working parliamentary majority.
The Democratic Party (PD), the big loser of the election after its centre-left coalition gained just under 23 percent of the vote, has been courted by the M5S. The League has refused categorically to work with the PD.
The PD is deeply divided between those who prefer to remain in opposition rather than prop up a Di Maio government -- a faction led by former prime minister Matteo Renzi -- and those who want dialogue to stop the far-right League gaining power.
The issue may not be decided until April 21, when the PD will choose a new leader after Renzi's post-election resignation.
The M5S chief whip in the Chamber Giulia Grillo told Il Fatto Quotidiano newspaper on Thursday that "up to now the PD has not wanted any dialogue".
On Friday Mattarella will meet the speakers of both houses of the Italian parliament and his predecessor Giorgio Napolitano.
Mattarella could give one of the trio an exploratory mandate to try to move the discussions forward, or plan a third round of consultations.
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