AbstractFollowing the adoption of the Post-2020 Kunming-Montreal Global Biodiversity Framework (KM-GBF) of the Convention on Biological Diversity (CBD) there is a clear science-policy need to protect habitat connectivity and track its change over time to safeguard biodiversity and inform conservation planning. In response to this need we describe an analytical, multi-indicator and multispecies approach for the rapid assessment of habitat connectivity at fine spatial grain and at the extent of an entire ecoregion. Out of 68 connectivity indicators we found through a literature review, we identified a key-set of six indicators that align with the Essential Biodiversity Variables framework and are suitable to guide rapid action for connectivity and conservation targets in the KM-GBF. Using these selected indicators, we mapped and evaluated connectivity change from 2011 to 2021 across the ecoregion of the St-Lawrence Lowlands in Quebec (~ 30,000 km2) for seven ecoprofile species representing regional forest habitat needs. For most of these species, trends over the last decade indicate a decline in effective connected area and metapopulation carrying capacity, via a division of large contiguous habitat into smaller fragments, whereas on average, habitat area slightly increased. These results highlight that temporal changes in habitat area and connectivity are not necessarily correlated and the urgent need to conserve and restore connectivity to meet targets under the KM-GBF. We provide an R-tool to support our general approach, which enables a comprehensive evaluation of connectivity for regional spatial planning for biodiversity in regions with moderate to high human disturbance.