Detectives from Victoria Police's major collision unit are investigating the crash at Diamond Creek, in Melbourne's outer northeast.
Emergency services were called to Chute Street about 10pm on Monday after reports of a collision between a 4WD and an electric motorcycle.
The rider of the bike, a 14-year-old boy from Diamond Creek, was treated at the scene but could not be revived.
The vehicle's driver, a 22-year-old from Eltham North, is assisting police as officers investigate the circumstances surrounding the collision.
The boy's death is the latest in a spate of fatal e-bike crashes across the nation, which have triggered calls for a safety crackdown on the popular means of transport.
In Sydney, an e-bike rider died a week earlier after being hit by a garbage truck. He was not wearing a helmet.
On the Sunshine Coast, a 15-year-old boy was charged over an e-bike crash that killed an eight-year-old boy in October.
State politicians have been pleading with the federal government to clamp down on the import and sale of e-bikes able to reach dangerously high speeds amid a spate of crashes.
On Tuesday, the Victorian Automotive Chamber of Commerce (VACC) criticised the state governments decision to reject a coroner's recommendation for e-bike registration.
In July 2025, Coroner Audrey Jamieson recommended the government consider methods to identify non-compliant e-bicycles, including requiring the registration of all e-bicycles following her investigation into the death of Nitin Haldipur Prabhu, who was riding a non-compliant e-bike.
However, in formal responses to the Coroners Court, both Victoria Police and the Department of Transport and Planning (DTP) rejected the implementation of an e-bike registration scheme.
In her response, DTP Acting Executive Director Fiona Green stated that registration "is not being actively considered at this time" because it "would be costly and may be a major disincentive to riding, consequently discouraging active transport".
VACC CEO Peter Jones on Tuesday said the decision represented a concerning reluctance to act on a coroners safety recommendation.
"Victoria has always led the nation on road safety initiatives. We shouldn't be waiting for other states when we have a clear recommendation from a coroner," he said.
The Chamber's research partnership with Monash University recently documented the scope of non-compliant e-bike use in Melbourne's CBD.
The Monash University study, published in August, found many e-bike delivery riders in Melbourne were breaking the law, including some reaching speeds of up to 54km/h.
Most of the 27,000 e-bikes observed were not using pedals, indicating they were throttle-powered or illegally modified, according to the research.
It urged authorities to look at the legal definition of e-bikes, enforce existing road rules for non-compliant devices, and close import loopholes.
WeRide, a group representing bicycle and e-bike companies, said leadership from the federal government was needed to ensure e-bikes entering Australia met local standards.
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