Biodiversity information is essential for understanding and managing the environment. However, identifying and providing the forms and types of biodiversity information most needed for research and decision-making is a significant challenge. While research needs and data gaps within particular topics or regions have received substantial attention, other information aspects such as data formats, sources, metadata, and information tools have received little. Focusing on the US southeast, a region of global biodiversity importance, this paper assesses the biodiversity information needs of environmental researchers, managers, and decision makers. Survey results of biodiversity information users' information needs, information-seeking behaviors and preferred information source attributes support previous conclusions that useful biodiversity information must be easily and quickly accessible, available in forms that allow integration and visualization and appropriately matched to users' needs. Survey results concerning additional information aspects suggest successful participation in both the creation and provision of biodiversity information include an increased focus on information search and other tools for data management, discovery, and description.