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A major challenge for biodiversity conservation is to mitigate the effects of future environmental change, such as land use, in important areas for biodiversity conservation. In the United States, recent conservation efforts by The Nature Conservancy and partners have identified and mapped the nation's Areas of Biodiversity Significance (ABS), representing the best remaining habitats for the full diversity of native species and ecosystems, and thus the most important and suitable areas for the conservation of native biodiversity. Our goal was to understand the potential consequences of future land use changes on the nation's ABS, and identify regions where ABS are likely to be threatened due to future land use expansion. For this, we used an econometric-based model to forecast land use changes between 2001 and 2051 across the conterminous U. S. under alternative scenarios of future land use change. Our model predicted a total of similar to 100,000 to 160,000 km(2) of natural habitats within ABS replaced by urban, crop and pasture expansion depending on the scenario (5% to 8% habitat loss across the conterminous U.S.), with some regions experiencing up to 30% habitat loss. The majority of the most threatened ABS were located in the Eastern half of the country. Results for our different scenarios were generally fairly consistent, but some regions exhibited notable difference from the baseline under specific policies and changes in commodity prices. Overall, our study suggests that key areas for conserving United States' biodiversity are likely threatened by future land use change, and efforts trying to preserve the ecological and conservation values of ABS will need to address the potential intensification of human land uses.