Abstract To explore the role of brain tyramine in reproductive worker honeybees, its effects after injection and oral treatment on brain dopamine levels and ovarian development in queenless worker honeybees are determined. Both tyramine injection and oral treatment in 10‐day‐old queenless bees leads to tyramine transportation into the brain and significantly elevates brain dopamine levels as a function of the tyramine concentration. Ovarian diameters are significantly larger in 10‐day‐old queenless bees treated with tyramine compared with queenless bees of the same age without tyramine treatment. Results on yolk formation in the ovary support the finding of increased ovarian diameter, suggesting that oral tyramine treatment accelerates ovarian development through dopamine effects and/or direct effects of tyramine on the ovary in queenless bees. Thus, tyramine has potential effects on the enhancement of brain dopamine levels and the acceleration of ovarian development for the transition of normal workers to reproductive worker honeybees.