In this publication, take stock of the knowledge and know-how induced by the BIODEV2030 project’s first step, which is to assess the main threats to biodiversity and provide elements to address the following specific questions: how were main threats to biodiversity identified and ranked and associated sectors selected in each country? Were the different methods converging in identifying top threats? What are their respective strengths, limitations and complementarities? What recommendations can we make to other actors (governments, NGOs, donors such as multilateral or bilateral development banks) wanting to follow similar approaches with the private sectors at national level? In each BIODEV2030 country, three main methods were typically combined to identify and rank main threats to biodiversity and then select (sub-)sectors: literature and data sources review, Species Threat Abatement and Restoration (STAR) metric and national experts’ elicitation. The results of the assessment and its processes led to a list of 12 key recommendations.