Abstract Background Sesame is an ancient oil crop that has been cultivated for centuries. It is an important source worldwide for food, industry and edible oil. Sesame plants are attacked by many pathogens during different stages of growth. Macrophomina phaseolina is considerable one of the most dangerous fungi that attacks sesame plants during their different growth stages. The impact of Streptomyces violaceoruber and Streptomyces hirsutus in comparison with Topsin-M fungicide on damping-off and charcoal rot caused by M. phaseolina and also on the growth of sesame plants was studied. Results In general, S. hirsutus was effective more than S. violaceoruber. S. hirsutus reduced the linear growth of M. phaseolina in vitro by 70.83%, while reduction with S. violaceoruber reached 53.89%. S. hirsutus isolate reduced pre-, post-emergence damping-off, and charcoal rot incidence by 58.3, 56.6, and 50%, under greenhouse conditions, in 2021 growing season, while the percentage reduction of disease was 43, 56.4, and 71.2% for 2021 and 2022 growing seasons, respectively, under field conditions at Fayoum Governorate, Egypt. Streptomyces treatments increased concentration of nitrogen, phosphorus, and potassium (NPK) in plant leaves, seed yield, and seed oil concentration more than Topsin-M treatment and untreated plants. Conclusions S. violaceoruber and S. hirsutus have proven that they can be used to combat soil-borne diseases, as well as improve growth parameters and increase yields.