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The utilization of heterosis is consided to be one of the most effective ways to increase crop yield.Oil rape is of obvious heterosis for biomass and seed yield.BnRabGDI3 is a gene differentially expressed in the sterile and fertile anther of B.napus two type lines J03AB(Msmsrfrf * msmsrfrf).BnRabGDI3 was cloned and compared with AtRabGDI3 by bioinformatics analysis.In this study,amiRNA interference technology was used to study the function and molecular mechanisms of BnRabGDI3.Most of transgenic plants displayed complete or partial male sterility: shrinking anthers with no pollen or with dramatically reduced microspores and significantly reduced number of pods.AtRabGDI3 expression in the transgenic plants was effectively silenced by 50-80%as analyzed by Q-PCR.Promoter was cloned and analysis by GUS staining which shows that the promoter of BnRabGDI3 expressed in the pollen specifically.Scanning and transmission electron microscopy demonstrated that aborted microspores were stuck together by exine.Based on the above results,we concluded that RabGDI3 is a new functional gene which regulates microspore development.These studies open a new window for gametogenesis regulation.