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Large, well described gaps exist in both what we know and what we need to know to address the biodiversity crisis. Artificial intelligence (AI) offers new potential for filling these knowledge gaps, but where the biggest and most influential gains could be made remains unclear. To date, biodiversity-related uses of AI have largely focused on tracking and monitoring of wildlife populations. Rapid progress is being made in the use of AI to build phylogenetic trees and species distribution models. However, AI also has considerable unrealized potential in the re-evaluation of important ecological questions, especially those that require the integration of disparate and inherently complex data types, such as images, video, text, audio and DNA. This Review describes the current and potential future use of AI to address seven clearly defined shortfalls in biodiversity knowledge. Recommended steps for AI-based improvements include the re-use of existing image data and the development of novel paradigms, including the collaborative generation of new testable hypotheses. The resulting expansion of biodiversity knowledge could lead to science spanning from genes to ecosystems — advances that might represent our best hope for meeting the rapidly approaching 2030 targets of the Global Biodiversity Framework.