玫瑰叶片离体培养胚状体的发生

Somatic Embryogenesis from the Leaves of Rosa rugosa in vitro Culture

[目的]为玫瑰转基因提供基础物质。[方法]以玫瑰试管苗叶片为外植体,以MS为基本培养基,研究了不同浓度6苄-基腺嘌呤(6-BA),腺嘌呤硫酸盐(Ad),萘乙酸(NAA)及其组合对胚状体发生的影响,同时也进行了不同蔗糖浓度对胚状体形成的研究。[结果]6-BA对胚状体的诱导效果较Ad好,NAA与二者配合使用可提高胚状体的发生率,其最佳浓度组合为6-BA 0.4 mg/L+Ad 0.2 mg/L+NAA 0.2 mg/L。蔗糖20～30 g/L有利于胚状体的形成。其组织学观察表明,胚状体起源于叶片气孔附近的上表皮细胞或上表皮下方的叶肉组织细胞,为单细胞起源。[结论]玫瑰叶片诱导胚状体的最佳培养基为:1/2 MS+6-BA 0.4 mg/L+Ad 0.2 mg/L+NAA 0.2mg/L+蔗糖20～30 g/L。

[ Objective ] The study was to provide-the fundamental material for transgenic research of Rosa rugosa. [ Method ] The leaves of Rosa rugosa plantlets were used as explants and cultured in MS basic medium. The effects of different concentrations of three kinds of plant growth regulators （6-BA, Ad, NAA） and their combinations on somatic embryogenesis were studied in vitro culture. Meanwhile, the influences of different concentrations of sucrose on somatic embryogenesis were studied. [ Result ] The induction effects of cytokinins 6-BA was better than Ad in embryoid induction. The auxin NAA combined with given concentrations of 6-BA and Ad could effectively raise the frequency of somatic embryogenesis. The optimum plant growth regulator combination for somatic embryogenesis was 0.4 mg/L 6-BA ＋ 0.2 mg/L Ad ＋ 0.2 mg/L NAA. 20 - 30 g/L sucrose added in the medium was beneficial to somatic embryogenesis. The results of histological observations indicated that somatic embryos originated from upper epidermis and mesophyll histiocyte single cell nearby stoma. [ Conclusion] The optimum medi- um for the somatic embryogenesis from leaf explants of Rosa rugosa was 1/2 MS ＋ 0.4 mg/L 6-BA ＋ 0.2 mg/L Ad ＋ 0.2 mg/L NAA ＋ 20 - 30 g/L sucrose.