Biodiversity supports sustainable food production, although recognition of its roles has been relatively neglected in the sustainable intensification literature. In the current study, the roles of biodiversity in sustainable food production are considered, assessing how these roles can be measured, the current state of knowledge and opportunities for intervention. The trajectory of global food production, and the challenges and opportunities this presents for the roles of biodiversity in production, are also considered, as well as how biodiversitybased interventions fit within wider considerations for sustainable food systems. The positive interactions between a diverse array of organisms, including annual crops, animal pollinators, trees, micro-organisms, livestock and aquatic animals, support food production globally. To support these interactions, a range of interventions related to access to materials and practices are required. For annual crops, major interventions include breeding crops for more positive crop–crop interactions, and the integration of a wider range of crops into production systems. For animal pollinators, major interventions include the introduction of pollinator populations into production landscapes and the protection and improvement of pollinator habitat. For trees, a major required intervention is the greater integration of perennial legumes into farmland. For micro-organisms, the implementation of agronomic practices that support beneficial crop-microbe interactions is crucial. For livestock production, breed and crop feedstock diversification are essential, and the implementation of improved methods for manure incorporation into cropland. Finally, in the case of aquatic production, it is essential to support the wider adoption of multi-trophic production systems and to diversify crop- and animal-based feed resources. These and other interventions, and the research needs around them, are discussed. Looking to the future, understanding the drivers behind trends in food systems is essential for determining the options for biodiversity in supporting sustainable food production. The increased dominance of a narrow selection of foods globally indicates that efforts to more sustainably produce these foods are crucial. From a biodiversity perspective, this means placing a strong emphasis on breeding for resource use efficiency and adaptation to climate change. It also means challenging the dominance of these foods through focusing on productivity improvements for other crop, livestock and aquaculture species, so that they can compete successfully and find space within production systems. New biodiversity-based models that support food production need not only to be productive but to be profitable. Thus, as well as describing appropriate production system management practices that enhance production and support the environment, the labour, knowledge, time required to operationalize, and other costs of new production approaches, must be considered and minimized. To support the future roles of biodiversity in sustainable food production, we recommend that particular attention be given to the longitudinal analysis of food sectors to determine how the diversity of foods consumed from these sectors has changed over time. Analysis is already available for crops, but related research is needed for livestock and aquaculture sectors. This analysis will then support more optimal cross-sectoral interactions, in terms of the contributions each sector provides to supplying the different components of human diets. Additional meta-analyses and synthetic reviews of case studies are required as an evidence base for biodiversity-based food production system interventions, but future studies should pay more attention to articulating the potential biases in case study compilation (the problem of ‘cherry picking’ positive examples) and the measures that have been taken to minimize such effects.