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Abstract: The energy fuels used for in the Greek transport sector are made up of gasoline consumed by automobiles, diesel oil consumed by taxis, trucks, maritime transport and railroads, and jet fuel used in the aircrafts. All these fuels are hydrocarbons that emit great amounts of CO2 which has a major impact in the global warming phenomenon. The issues relating to climate change, the soaring energy prices, and the uncertainty of future oil supplies, have created a strong interest in alternative transportation fuels. During the past decade biofuels in the form of blended gasoline and biodiesel have begun to find place in energy economy. The Greek car market shows a remarkably low rate in the penetration of biodiesel compared to the average European Union market. This work compares the environmental impacts of the use of gasoline, diesel and biodiesel in Greece using as a tool for the comparison the Life Cycle Assessment (LCA) methodology. The environmental impacts taken into consideration include: organic respiratory effects, inorganic respiratory effects, fossil fuels, acidification – eutrophication, greenhouse effect, ecotoxicity and carginogenic effects. From the environmental point of view, biodiesel appears attractive since its use results in significant reductions of GHG emissions in comparison to gasoline and diesel. It also has lower well-to-wheel emissions of methane. However, the use of biodiesel as transportation fuel increases emissions of PM10, nitrous oxide, nitrogen oxides (NO x ) as well as nutrients such as nitrogen and phosphorous; the latter are the main agents for eutrophication. This study can be considered as an opportunity for further research and evaluate the available options for a sustainable transportation system planning in Greece. [Copyright &y& Elsevier]