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• Comprehensive phenology from the onset of the primordial bud to the senescence phase, is documented for Macrotyloma uniflorum. • Floral organs exhibits ultrastructural variation in Macrotyloma uniflorum. • Macrotyloma uniflorum accessions from Republic of Benin are non-echinate; other accessions exhibit spinuliferous pollen. • There is a positive correlation between days to flowering and most floral traits. • Pollens of Macrotyloma uniflorum are viable during the day. The absence of information on the reproductive biology of some nutritive legumes has been an obstacle to their genetic improvement. Gathering information on reproductive biology contributes significantly to the interest of breeders to have adequate tools for initiating the improvement program through crosses among high performing genotypes. To contribute to this, we examined the flowering phenology, pollen biology, stigma receptiveness at bud dehiscence and anthesis phases, and the in-vitro germination of the pollens in horsegram (Macrotyloma uniflorum). Pollens of horsegram were found to be either tricolporate or triporate, echinates in all accessions except in the collections from Benin. Ovaries in horsegram showed great variation in the number of ovules and activities of peroxidase indicated the receptiveness of stigma at bud dehiscence phase and during anthesis. A high number of ovules per ovary was found in accessions from Australia (Leichardt 1 and 2), Mali (LeichardtMali) and India (CPI22679). Pollen viability analysis implies the possibility of better results when pollinating during the day and immediately after the flower opens. The study provides information on the reproductive biology of horsegram for future hybridization program in order to enhance the performance of the crop and increase production. It also reveals differences in accessions from Benin compared with all other accessions from Mali, India or Australia. [ABSTRACT FROM AUTHOR]