Novel fire regimes are emerging worldwide and pose substantial challenges to biodiversity conservation. Addressing these challenges and mitigating their impacts on biodiversity will require developing a wide range of fire management practices. In this paper, we leverage research across taxa, ecosystems and continents to highlight strategies for applying fire knowledge in biodiversity conservation. First, we define novel fire regimes and outline different fire management practices in contemporary landscapes from different parts of the world. Next, we synthesize recent research on fire use and biodiversity, and provide a decision-making framework for biodiversity conservation under novel fire regimes. We recommend that fire management strategies for preserving biodiversity should consider both social and ecological factors, iterative learning informed by effective monitoring, and developing and testing new management actions. An integrated approach to learning about fire and biodiversity will help to navigate the complexities of novel fire regimes and preserve biodiversity in a rapidly changing world.