More than half of the global emissions due to energy use in agriculture derives from the combustion of fossil fuels to power tractors and machineries. Paying attention to energy sources and fossil fuels use in agriculture could offer significant opportunities for on-farm mitigation actions directly focused on CO2. With this regard, the reduction of fossil fuels, and the increasing use of energy sources which are not petroleum-based, are key factors for the development of energy security and the achievement of carbon emissions reduction in agriculture. In recent years, agricultural machinery manufacturers started investing in solutions powered by renewable energy sources and a first-generation of tractor prototypes fueled by e-fuel, biomethane or hydrogen, are currently under development. The present study aimed to investigate the actual adoption and intention to adopt alternative fuels to power agricultural machinery among Italian farmers, analyzing also the perceived barriers to adoption and their relationships with participants' sociodemographic characteristics. A questionnaire was administered to a sample of 409 Italian farmers asking them to indicate between e-fuel, biomethane and hydrogen which is the best alternative to fossil fuels in agriculture. Then, they were asked to rate their intention to adopt the selected energy source to power their machinery and the importance of barriers to adoption (costs, performance, safety, autonomy, refueling). Finally, they were asked if they had experience with agricultural machinery powered by alternative fuels. Results showed that the interest in using alternative energy machinery was higher for biomethane and e-fuel. Costs and refueling were perceived as themain barriers to adoption; Safety was indicated as a critical issue forhydrogen, while autonomy was relevant for e-fuel and biomethane. Furthermore, safety and refueling results correlated with the years of education. Implications for targeted interventions to support a wider adoption of these alternative fuels are discussed.