Summary: Effective natural resource management requires knowledge exchange between researchers and managers to support evidence‐based decision‐making. To achieve this, there is a need to align research with management and policy needs. This project aimed to identify the flow‐related ecological knowledge needs for freshwater fish to better inform environmental water management in the Murray–Darling Basin, south‐eastern Australia. Our major objective was to provide an up‐to‐date assessment of scientific research and integrate this with the knowledge requirements of relevant managers to guide future research. We reviewed the contemporary scientific literature and engaged managers specifically responsible for delivering flows for fish outcomes via a questionnaire and workshop. Research on fishes of the MDB has generally evolved from single locations and/or times to larger spatio‐temporal scales, including multiple sites, rivers and catchments. There has also been a trend from single life stage studies to incorporation of multiple life stages and population processes. There remain, however, significant deficiencies in knowledge for most native species, many of which are threatened. Four agreed key knowledge gaps were derived from the literature review and managers' suggestions: (i) population dynamics, (ii) movement, dispersal and connectivity, (iii) survival and recruitment to adults and (iv) recruitment drivers. To inform policy and management, managers desired timely advice, based on robust research and monitoring. Fish species of most relevance to managers were those highly regarded by community stakeholders and whose life histories and population dynamics are potentially influenced by flow. Populations of these mostly large‐bodied, angling species (e.g. Murray Cod, Golden Perch and Silver Perch) have declined, often due to river regulation and, in conjunction with managers' priorities, are relevant candidates for research to support the management of flow to rehabilitate fish populations in the MDB. [ABSTRACT FROM AUTHOR]