Abstract: The effects of different factors on the callus induction and plant regeneration from mature embryos of 27 winter wheat genotypes of Zhoumai were studied. The shoot differentiation rate of callus from mature embryos with cross cutting was 66.7%, 2.1 and 4.3 times higher than that from fragmented embryos and whole embryos, respectively. Although Dicamba and 2,4-D induced callus from explants with high frequency (96.3%-97.7%), Dicamba was more effective for the induction of embryogenic callus, while the callus differentiation rate from 2, 4-D was 4.0 times greater. When the concentration of Dicamba was 2.0-4.0 mg/L, the rate of callus induction and differentiation were both higher. Supplement of additional cytokinins to callus differentiation medium had different effects on the rate of callus differentiation. The best rate of shoot formation was observed with addition of 2.0 mg/L 6-BA (45.8%). When contact of explants with callus induction medium was two or three weeks the rate of shoot formation was high. Callus drying treatment of 6 h increased the differentiation rate slightly, but there was no significant difference compared with no desiccation.