The standings leader started from pole but was required to fend off a strong challenge from Yamaha’s Jorge Lorenzo, who finish second.
Valentino Rossi was able to secure a podium place on his 300th career race but the stage belonged to Marquez as he picked up his first win in Mugello.
Andrea Iannone made much of the early running but was quickly reeled in by Lorenzo.
Marquez was the next to overtake Iannone but he was made to toil for the lead and did not edge ahead until the 17th lap and, even then, the pair regularly swapped places over the next few circuits.
Having regained the lead by the final lap, Repsol Honda’s Marquez was able to hold off Lorenzo and move to the 150-point mark for the season – 53 clear of Rossi in second.
It was also enough to deny six-time MotoGP world champion Rossi a 107th career win in his landmark race on home soil.
Dani Pedrosa was able to salvage fourth place while early pacesetter Iannone slipped to seventh by the end of the race.
The only sour note of the race was a collision between Cal Crutchlow and Stefan Bradl, although both were later confirmed to be unharmed.
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