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The effects of the microspore developmental stage, hormones and culture condition on anther in vitro culture of lily (Lilium spp.) were discussed. The results showed that when the flower buds were about 23-26 mm long, the microspores were at the uninucleate stage wluch was suitable for culture and the culture under the darkness would promote the callus induction of anther. The induction frequency could reach 42.5% in the optimized medium which was MS+[6-BA(0.5)+KT(2.0)+2,4-D(1.0)]mg·L-1.The rate of callus differentiation could reach 31.57% in the optimized medium wluch was MS+NAA(1.5,2.0)mg·L-1.