The M5S rules the country in coalition with Matteo Salvini’s far-right League, which won a majority in Italy’s EU parliamentary vote on the back of a nationalist “Italians First” campaign.
The confidence vote, which poses the question: “Do you confirm Luigi Di Maio as the political head of the M5S?” will take place on Thursday on the party’s online platform, Rousseau.
“You decide. I’m asking to put my role as party leader to the vote,” Di Maio said on the platform on Wednesday.
“If the Movement renews its faith in me, we’ll get to work... with even more commitment and dedication.”
The EU vote results confirmed a change in the balance of power between the ruling parties, with M5S - which polled 32.5 per cent at the general election in 2018 - garnering just 17 per cent on Sunday compared to the League’s 34 per cent.
Di Maio has blamed the M5S’ poor performance on low voter turnout, as well as a mud-slinging campaign by the League against which it was slow to retaliate.
In the event of Di Maio being ousted as leader of the M5S, it is unclear who would replace him.
M5S often turns to Rousseau to make decisions and craft policy.
The last vote was in January, when members chose to block a criminal case against Matteo Salvini, the co-deputy prime minister and leader of the League, for refusing to allow migrants to disembark from a rescue ship.