Background There is a good knowledge of the existing fauna and flora in the Portuguese region of Alentejo. However one of the main transport and energy corridors linking Portugal to Spain lies in this region. This puts significant pressure on the area’s wildlife, as the mortality data for the linear infrastructure (roads and power lines) shows. Within this socioeconomic context, stakeholders and public and private bodies have actively participated with universities to solve the environmental problems caused by transport and energy networks. The situation for wildlife could be improved through a coordinated strategy, based on research done at the University of Évora in the field of functional and structural connectivity mapping, which takes into account the movements and habitat use/preferences of target fauna species. Objectives The main objectives of the LIFE LINES project are to evaluate and disseminate practices to mitigate the negative effects that transport and energy infrastructure have on wild fauna, and to simultaneously promote the creation of a demonstrative ‘green Infrastructure’ based on ecological corridors and stepping stones. Running alongside grey infrastructure e.g. roads, railways and energy networks, the enhanced green infrastructure will improve connectivity and the conservation of local and regional biodiversity. A green infrastructure strategy will be used, for the first time, as a decision-support tool for practical conservation aims. Expected results: The LIFE LINES project will implement actions and test solutions to make roads, abandoned railways and power lines (medium and high voltage) more sustainable for wildlife, by managing this infrastructure within the ‘green infrastructure’ concept. Specifically, the project is expected to: Increase landscape connectivity in the study area and with surrounding areas, and reduce wildlife mortality caused by grey infrastructure; Create a network of corridors and micro-reserves in the vicinity of linear infrastructure, including road embankments and around utilities’ pylons; Promote practices for the rapid detection and control of invasive alien plant species in grey infrastructure; Create a nursery and seed mixtures of native plants for use in habitat creation, recover of areas invaded by exotic flora, and create biodiverse flora communities within micro-reserves; Create a national wildlife mortality database that can be used as a common platform for all infrastructure operators and nature conservation institutions; Increase people’s awareness of, and get their input on the problem of wildlife mortality from vehicle collisions, through the development of a mobile application (tablet and smartphone); Test the efficiency of new designs of landing deterrents mounted on pylons of medium and high voltage power lines; and Test the efficacy of various ‘self-learning’ machines/ systems for the identification of amphibian and bird roadkill.