BackgroundCurrent recommendations for physical exercise include information about the frequency, intensity, type, and duration of exercise. However, to date, there are no recommendations on what time of day one should exercise. The aim was to perform a systematic review with meta-analysis to investigate if the time of day of exercise training in intervention studies influences the degree of improvements in physical performance or health-related outcomes.MethodsThe databases EMBASE, PubMed, Cochrane Library, and SPORTDiscus were searched from inception to January 2023. Eligibility criteria were that the studies conducted structured endurance and/or strength training with a minimum of two exercise sessions per week for at least 2 weeks and compared exercise training between at least two different times of the day using a randomized crossover or parallel group design.ResultsFrom 14,125 screened articles, 26 articles were included in the systematic review of which seven were also included in the meta-analyses. Both the qualitative synthesis and the quantitative synthesis (i.e., meta-analysis) provide little evidence for or against the hypothesis that training at a specific time of day leads to more improvements in performance-related or health-related outcomes compared to other times. There was some evidence that there is a benefit when training and testing occur at the same time of day, mainly for performance-related outcomes. Overall, the risk of bias in most studies was high.ConclusionsThe current state of research provides evidence neither for nor against a specific time of the day being more beneficial, but provides evidence for larger effects when there is congruency between training and testing times. This review provides recommendations to improve the design and execution of future studies on this topic.Registration: PROSPERO (CRD42021246468).Supplementary InformationThe online version contains supplementary material available at 10.1186/s40798-023-00577-5.