木蓝愈伤组织诱导与不定芽分化培养

Indigofera bungeana walp.:Callus Induction and Adventitious Bud Differentiation Culture

初步建立了木蓝(Indigofera bungeana Walp.)的离体再生培养体系,为其基因工程育种研究提供前期技术参考。通过种子无菌播种技术获得野生木蓝无菌实生苗,以胚轴、茎段和叶片为外植体,筛选木蓝愈伤组织诱导、不定芽分化、增殖、壮苗培养的最适培养基配方。结果显示,木蓝种子无菌播种发芽率达98%,最适培养基为1/2MS。3种外植体在不同培养基下均能诱导出愈伤组织,根据愈伤组织生长及质地色泽筛选出最适诱导培养基,其中,叶片为MS+0.5 mg/L 6-BA+0.5 mg/L 2,4-D+0.5 mg/L NAA;茎段为MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+0.1 mg/L NAA;胚轴为MS+0.1 mg/L 6-BA+0.1 mg/L2,4-D+0.5 mg/L NAA。相比较叶片和茎段,胚轴来源的愈伤组织适于不定芽分化,最适分化培养基为MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+0.1 mg/L NAA和MS+2.0 mg/L 6-BA。不定芽增殖培养基为MS+1.0 mg/L 6-BA+0.1 mg/L NAA,壮苗和生根的最适培养基均为1/2MS。

The system of in vitro regeneration of Indigofera bungeana walp.was established in order to provide a preliminary experimental basis for genetic-engineering breeding of Indigofera bungeana walp..Aseptic seedlings of Indigofera bungeana walp.were obtained through aseptic seeding.Using plumular axis,stem segments and leaves as explants,the optimum medium formula was selected for callus induction,adventitious bud differentiation,propagation and elongation.The results showed that the germination rate of the Indigofera bungeana walp.seeds reached 98%on the 1/2MS medium.The three explants could be induced callus in different media,and the suitable medium was selected according to the growth,texture and color of callus.For the leaves,the optimum medium was MS+0.5 mg/L 6-BA+0.5 mg/L 2,4-D+0.5 mg/L NAA,for the stem segment,the optimum medium was MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+0.1mg/L NAA,for the plumular axis,the optimum medium was MS+0.1 mg/L 6-BA+0.1 mg/L 2,4-D+0.5 mg/L NAA.Compare with stem segments and leaves,the callus from plumular axis were suitable for adventitious bud differentiation,and the optimum medium were MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+0.1 mg/L NAA and MS+2.0 mg/L 6-BA.The adventitious bud propagation medium was MS+1.0 mg/L 6-BA+0.1 mg/L NAA,and the optimum medium for elongation and rooting was 1/2MS.