Abstract:This paper first reported the dynamic development of male gametophyte in autotetraploid broccoli using Ehrlichps hematoxylin staining-methyl salicylate clearing technique. Results showed that the male gametophyte in autotetraploid broccoli shared the similar development with that in diploid broccoli, which could give rise to one vegetative cell and one generative cell by one miotic division, while the generative cell could divide into two sperms by the othermiotic division and the mature pollen was 3-celled with four or three apertures. Abnormality appeared mainly in uninuclete stage of male gametophyte development with an average frequency of 12.14%.