Background: Environmental issues, e.g. climate change, fossil resource depletion have triggered ambitious national/regional policies to develop biofuel and bioenergy roles within the overall energy portfolio to achieve decarbonising the global economy and increase energy security. With the 10 % binding target for the transport sector, the Renewable Energy Directive confirms the EU's commitment to renewable transport fuels especially advanced biofuels. Imola is an elite poplar clone crossed from Populus deltoides Bartr. and Populus nigra L. by Research Units for Intensive Wood Production, Agriculture Research Council in Italy. This study examines its suitability for plantation cultivation under short or very short rotation coppice regimes as a potential lignocellulosic feedstock for the production of ethanol as a transport biofuel. A life cycle assessment (LCA) approach was used to model the cradle-to-gate environmental profile of Imola-derived biofuel benchmarked against conventional fossil gasoline. Specific attention was given to analysing the agroecosystem fluxes of carbon and nitrogen occurring in the cultivation of the Imola biomass in the biofuel life cycle using a process-oriented biogeochemistry model (DeNitrification-DeComposition) specifically modified for application to 2G perennial bioenergy crops and carbon and nitrogen cycling. Results: Our results demonstrate that carbon and nitrogen cycling in perennial crop-soil ecosystems such as this example can be expected to have significant effects on the overall environmental profiles of 2G biofuels. In particular, soil carbon accumulation in perennial biomass plantations is likely to be a significant component in the overall greenhouse gas balance of future biofuel and other biorefinery products and warrants ongoing research and data collection for LCA models. We conclude that bioethanol produced from Imola represents a promising alternative transport fuel offering some savings ranging from 35 to 100 % over petrol in global warming potential, ozone depletion and photochemical oxidation impact categories. Conclusions: Via comparative analyses for Imola-derived bioethanol across potential supply chains, we highlight priority issues for potential improvement in 2G biofuel profiling. Advanced clones of poplar such as Imola for 2G biofuel production in Italy as modelled here show potential to deliver an environmentally sustainable lignocellulosic biorefinery industry and accelerate advanced biofuel penetration in the transport sector.