Championship leader Maverick Viñales led qualifying on Saturday in Mugello, beating Yamaha team-mate Valentino Rossi by just two-tenths in a very close session.
The Spaniard beat Rossi on his home soil ahead of third-placed Ducati's Andrea Dovizioso, with Italians lining up second, third and fourth as Ducati stand-in Michele Pirro was just 0.043 off his team-mate, giving the locals something to cheer for.
The Honda pair of Dani Pedrosa and Marc Marquez complete the second row in fifth and sixth.
Heading up row three is one of Mugello’s most successful riders, Ducati's Jorge Lorenzo, ahead of compatriot Alvaro Bautista as the top independent rider for Aspar.
Pramac's Danilo Petrucci had seemed set for a front row start before having his fastest laptime cancelled for exceeding track limits, and the Italian dropped from a provisional P2 to take ninth on the grid at home – in front of a best ever for Marc VDS rider Tito Rabat to complete the top ten.
Yamaha Tech 3's Johann Zarco had a more difficult session to take eleventh, with Aleix Espargaro crashing out early and ending the session in P12.
It was a very close session with the top 12 covered by less than a second.
A Q1 susprise came when Cal Crutchlow failed to make the cut for LCR Honda and will line up 13th ahead of Hector Barbera and Jonas Folger, 15th.
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