三叶木通体细胞胚胎发生及植株再生研究

Somatic Embryogenesis and Plantlet Regeneration of Akebia trifoliate

  • 摘要: 以三叶木通成熟种子的幼胚为材料,探讨不同浓度2,4−D(1.0、2.0、4.0 mg/L)、NAA(0.05、0.1、0.2、0.5、1.0 mg/L)、6−BA(0.1、0.2、0.5、1.0 mg/L)、KT(0.2、0.5、1.0 mg/L)的植物生长调节剂对三叶木通愈伤组织的诱导及体细胞胚胎发生的影响,分别筛选出诱导愈伤组织、体细胞胚、次生胚增殖、植株再生的最适培养基,以此间接途径建立三叶木通体细胞胚胎发生及植株再生体系。结果表明:三叶木通种子最佳灭菌处理方式为0.1 % HgCl2灭菌18 min,无菌水冲洗干净后接种在添加了0.05 % PPM的愈伤诱导培养基上;三叶木通幼胚诱导愈伤组织的最佳培养基是 MS+2.0 mg/L 2,4−D+0.2 mg/L NAA,愈伤诱导率为95.67%;胚性愈伤诱导三叶木通体胚发生的最佳培养基为MS+0.1 mg/L NAA+0.2 mg/L 6−BA,体胚诱导率为55%;次生胚增殖的最佳培养基为MS+0.05 mg/L NAA+0.1 mg/L 6−BA,增殖倍数达19.84,体胚随继代次数增多逐渐分化,次生胚增殖活力变弱;成熟的体胚分化后一部分可再生成苗,在MS中能起到壮苗作用,再生植株转移至人工光照培养箱中炼苗3~5 d再转入V(泥炭土)∶V(蛭石)∶V(珍珠岩)为2∶1∶1的基质中能够稳定生根成活且形态正常。

     

    Abstract: The young embryos of the mature seed of Akebia trifoliate were used to study the effect of different concentrations of plant growth regulators: 2, 4−D (1.0, 2.0, 4.0 mg/L), NAA (0.05, 0.1, 0.2, 0.5, 1.0 mg/L), 6−BA (0.1, 0.2, 0.5, 1.0 mg/L), KT (0.2, 0.5, 1.0 mg/L) on callus induction and somatic embryogenesis of A. trifoliate. And select the medium for inducing callus, somatic embryo, proliferation of secondary embryos and plant regeneration. The system of embryogenesis and plant regeneration of A. trifoliate was established through indirect induction. The results show that the optimal sterilization treatment was 0.1 % HgCl2 for 18 min, washed with sterile water and inoculated on callus induction medium supplemented with 0.05 % PPM; the optimal medium for callus induction from immature embryos of A. trifoliate was MS supplemented with 2.0 mg/L 2, 4-D+0.2 mg/L NAA, and the callus induction rate was 95.67 %. The optimal medium for somatic embryogenesis was MS supplemented with 0.1 mg/L NAA+0.2 mg/L 6−BA, and the somatic embryo induction rate was 55%. The optimal medium for the proliferation of secondary embryos was MS supplemented with 0.05 mg/L NAA+0.1 mg/L 6-BA, and the proliferation ratio reached 19.84. Somatic embryos differentiated gradually with the increase of subculture times and the proliferation activity of secondary embryos became weak. Part of mature somatic embryos can be regenerated into seedlings after differentiation, the regenerated plants can grow strong in MS, and they were transferred to an artificial light incubator for 3–5 days, and then transferred to a 2∶1∶1 matrix of peat: vermiculite: perlite. The roots were stable and the morphology was normal.

     

/

返回文章
返回