The Maharashtra Navnirman Sena (MNS) chief Raj Thackeray chief will be leading the stir at the Vashi toll plaza in adjoining Navi Mumbai.
Police took MNS workers into custody at several places including Pune, Ahmednagar and Vasai.
Tyres were burnt in Thane, while in Pune MNS workers deflated tyres of some vehicles.
The stir has commenced despite police serving a notice to Raj warning against participating in the agitation.
Undeterred by the notice, Thackeray, whose supporters vandalised scores of toll plazas across the state over the last fortnight, said traffic on the state highways was stopped from 9 AM.
However, in some relief for commuters, Raj Thackeray had said yesterday that life in cities would not be affected. "We have decided to exclude schools and colleges from the stir in view of exams. The cities won't be affected by the stir," he said.
"A person from the state government was in touch with me on the phone requesting me to call off the stir," the MNS leader has said yesterday.
Maharashtra Police has decided to press into service contingents of Rapid Action Force and State Reserve Police to deal with the stir.
As a preventive measure, police stations across the state had served notices to MNS leaders and workers under section 149 of CrPC, which empowers police officers to take steps to prevent cognisable offences.