Carbohydrate sources are one of important factors associated with macro- and micro nutrients and phytohormones in vitro culture medium for shoot growth. The optimal carbohydrates for eight species of the genus Prunus which are economically important fruit crop was evaluated at the initiation and elongation stages. All carbohydrate seemed utilized for the bud break and leaf growth at the early stage of culture. However, shoot elongation and fresh weight of species tested were superior in the medium containing 90 mM of fructose or glucose rather than sucrose. There was no difference between glucose and fructose. Adventitious shoots from the axillary buds were induced in most species but no significant differences were observed except for two species (P. salicina ‘Shiro’ and P. tomentosa). These result demonstrated that glucose and fructose were suitable carbohydrate sources for diverse Prunus species than sucrose, although the response to the carbohydrates in the medium were slightly different in the species.