The factors inhibiting ovarian development and oviposition in worker honeybees Apis mellifera L. were investigated in four kinds of honeybee colonies; queen-caged colonies（tg1）, queenless colonies with broods（tg2）, queenless and broodless colonies（tg3） and normal queen colonies（CK）. Workers’ovarian development was observed under the microscope and a developmental index of oocyte development determined. The results show that, relative to CK, the degree of ovarian development was least in tg1 workers, higher in tg2 workers and greatest in tg3 workers. Differences in the ovarian development index were significant between the four groups after 31 days（P<0.05). The pre\|oviposition period of tg1, tg2 and tg3 workers were 35d, 22d and 17d, respectively. The comb capping time of tg2 and tg3 were 8d and 6d respectively. However, CK or tg1 workers did not cap combs, and CK workers didn’t oviposit. We conclude that the prolonged absence of a queen was more effective in triggering workers’ovarian development and oviposition than either a relatively short absence of a queen or an aging queen. The presence of a brood also inhibited worker’s ovarian development in queenless colonies, so addition of brood comb to such colonies could prevent workers from ovipositing.