Linear infrastructure networks such as roads, railways, navigation and irrigation canals, and power lines have grown exponentially since the mid-20th century. Most of these networks built before the 1990s have a significant impact on the environment. While there is no doubt that humanity needs infrastructure to ensure safe, secure and sufficient access to food, water and energy, it is essential to prevent the loss of biodiversity and ecosystems which are also at the basis of the provision of such fundamental services. Those complex, interconnected issues cannot be tackled without research and innovation, both in the fields of biodiversity and of infrastructure. IENE (Infrastructure Ecology Network Europe) was set up in 1996 to meet this need. Its mission is to promote the exchange of knowledge, experience and best practice in safe and sustainable pan-European transport infrastructure. With a status of an association today, this independent network has more than 400 members consisting of researchers, engineers, decision makers and infrastructure operators. IENE functions as an international and interdisciplinary forum. It supports cross-border cooperation in research, mitigation, planning, design, construction and maintenance in the field of biodiversity and transport infrastructure. Every two years, IENE organises an international conference to present cutting-edge research, identify pressing issues and problems, discuss effective solutions and map out future activities in the field of transport ecology and infrastructure. We are very glad to present you in this special issue some of the best scientific outcomes of the IENE 2020 conference, hoping that it will contribute to further breakthroughs in science and uptake in policy-making and practices on the ground. We commend the organising team of the University of Evora, Portugal, for their excellent programming of the conference and for having gathered exceptional scientists on the topic of biodiversity and infrastructure. They managed to host a high-quality event, despite the many adjustments that had to be done because of the covid-19, including postponing the conference to January 2021 and holding it entirely online. The topic of IENE conference 2020 was “Linear Infrastructure Networks with Ecological Solutions” and the motto was “working together”. This means that every stakeholder has a role to play, and that biodiversity should be considered at all governance scales and during all phases of the set-up of infrastructure. The papers selected here are of particular interest to follow the path set forth in the conference’s final declaration, that is included in this issue.