The long-awaited opening aims to bring an end to years of hazardous travel along the existing route, which has been plagued by accidents.
If owners fail to refute the evidence collected by AADE, they will be fined 10,000 euros, along with the payment of annual road taxes plus any late fees.
From Tuesday onwards, those crossing Central Greece will pay reduced tolls, ranging from 2.8 to 3 euros.