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Our planet is facing many complex environmental challenges, including the loss of biodiversity and rapidly changing climate conditions, driven by intensifying human-nature interactions worldwide.Dramatic advances in the biological sciences over recent years are made possible by new tools to study life at many scales, from identifying mutations in a single gene to monitoring changes in plants, animals, and microbes over an entire continent. These tools have the potential to usher in a new era of continental-scale biology (CSB) in which researchers can combine data from various realms across organizational, spatial, and temporal scales, addressing questions on biological processes and patterns that cannot be answered by observations at either small or large scales alone. This report, prepared at the request of the National Science Foundation, sets out a vision for the development of CSB and identifies the research areas that could most benefit from multi-scale approaches. Advancing the use of CSB to address a wide range of biological and societal challenges will require the development of integrated conceptual frameworks and theories to guide research, deployment of emerging technologies, and development of a skilled workforce to synthesize the vast amounts of data from various sources.