Abstract: ObjectiveFlowering habit of water chestnut and its fruiting characteristics after artificial supplementary pollination were studied, in order to provide references for crossbreeding of water chestnut. MethodUsing Guiti 2 as materials, the growth situation of stalk and inflorescence of water chestnut was observed, so as to understand flowering regulation of water chestnut and growth of stamen and pistil. And after artificial supplementary pollination and spontaneous pollination, the inflorescences were observed with microscope, respectively, and then the fruiting situations of water chestnut were compared. ResultThe results showed that, the stamen and pistil on the same scape reached peak when the lengths of scape were about 37.00 and 46.90 cm, respectively, both height difference was 9.84 cm, and time difference was 28.5 h. Furthermore, 9:00-11:00 am was the optimal period for artificial supplementary pollination, pistil instantly withered about 7.0 h after artificial supplementary pollination. The seed setting rates after artificial supplementary pollination and spontaneous pollination was 43.0%and 15.5%, respectively. ConclusionThe artificial supplementary pollination can significantly improve seed setting rates of water chestnut. Furthermore, there still exist time differences among characteristics, growth and full bloom of stamen and pistil, so the time difference was key factor for artificial supplementary pollination.