以'皖甜1号'甜叶菊花蕾为试验材料,利用正交设计L9(34),分别以MS、B5和N6为基本培养基,研究了不同浓度的NAA、6-BA以及蔗糖和低温预处理、暗处理时间等条件,对甜叶菊花药愈伤组织诱导的影响以及不同培养基对甜叶菊花药愈伤组织分化影响.试验得出甜叶菊花药愈伤组织最佳诱导的培养基配方为MS+3 mg/L 6-BA+0.5 mg/L NAA+60 g/L蔗糖+6 g/L琼脂,低温预处理1 d、暗培养14 d有利于甜叶菊愈伤组织的诱导,甜叶菊愈伤组织分化诱导的最佳培养基配方为MS+1 mg/L6-BA+0.2mg/L NAA+30g/L蔗糖+6 g/L琼脂,不定芽在不加任何植物生长调节剂的MS培养基上伸长后,接种在1/2MS+0.1 mg/L NAA+20g/L蔗糖+6 g/L琼脂培养基上生根效果好,生根率可达93.33％.该试验初步建立了甜叶菊花药离体培养技术体系,获得了甜叶菊花药培养再生植株,为甜叶菊新种质的创制和新品种的选育提供了帮助.“,”By using alabastrum of Stevia rebaudiana vr. 'wantian No.1' as test materials, the effects of different treatments including different concentrations of NAA, 6-BA and sucrose, cold pretreatment and dark treatment times on callus induction and callus differentiation of stevia anther and were studied in the basic mediums of MS, B5, and N6 by orthogonal design L9(34) respectively. The results showed that the best medium for stevia callus induction was selected as MS, 0.5 mg/L NAA, 3 mg/L 6-BA,60 g/L sucrose and 6 g/L agar. The stevia callus induction was also benefited after cold pretreating for 1 d and darkness culture for 14 d. And the optimum medium components for callus differentiation induction were selected as MS, 1 mg/L 6-BA, 0.2 mg/L NAA, 30 g/L sucrose and 6 g/L agar. After elongation on MS culture medium without any plant growth regulator, the induced adventitious buds were transferred into agar culture medium with 0.1 mg/L NAA, 20 g/L sucrose and 6 g/L agar, and showed good rooting effect which the rooting rate reached 93.33％. The regeneration system for anthers culture in vitro of Stevia rebaudiana was preliminary established in this study and anther culture regenerated plants were obtained, which might provide help for the creation and breeding of new germplasms of stevia.