葡萄柚不定芽再生体系建立研究

为建立葡萄柚上胚轴离体再生体系,且为遗传转化技术在葡萄柚上的应用提供产参考,以葡萄柚种子实生苗上胚轴为材料,研究了不同激素种类与浓度对上胚轴愈伤组织诱导与分化及无菌苗生根的影响。结果表明:在添加KT 1.5～2.0 mg/L的MT培养基上,上胚轴切口愈伤发生率及对应的不定芽发生率均高于对照及其它处理;在KT 2.0 mg/L的基础上,添加浓度为0.2～0.3 mg/L的NAA对愈伤分化无增加作用,但对不定芽分化率有较大的提高作用,不定芽分化率达87.23%;3种激素比较,ABT 1.0 mg/L的生根效果高于其它处理,生根率达81.3%;卡那霉素对上胚轴的选择压力为60mg/L。综上所述,MT基本培养基中添加KT 2 mg/L及NAA 0.2～0.3 mg/L对葡萄柚上胚轴的不定芽发生率有显著的促进作用;ABT是无菌苗生根的较合适激素;卡那霉素对上胚轴的选择压力为60 mg/L。

Study on Regeneration System of Adventitious Bud from Grapefruit

In order to construct the in vitro regeneration system of epicotyls from grapefruit,and to provide useful references for the application of genetic transformation technique in grapefruit,the author used the epicotyls of seedlings from grapefruit seeds as explants in tissue culture,and studied the effects of different hormone kinds and concentrations on the inducement and differentiation of epicotyls callus as well as the rooting of aseptic seedlings.The results showed that the callus occurrence rate of epicotyls and its adventitious bud occurrence rate in MT culture medium supplemented with KT 1.5 ～ 2.0 mg/L were higher than those in the control and other media.Adding NAA 0.2 ～ 0.3 mg/L into MT culture medium with KT 2.0 mg/L had no a promotive function on the callus differentiation,but had a great increasing function on the differentiation rate of adventitious buds,reaching 87.23%.Among three kinds of hormones,adding ABT 1.0 mg/L obtained higher rooting rate(81.3%) than that of the others.The selection pressure of Kanamycin to epicotyls was 60 mg/L.In conclusion,the addition of KT 2.0 mg/L and NAA 0.2 ～ 0.3 mg/L in MT basic culture medium could remarkably improve the differentiation rate of adventitious buds from the epicotyls of grapefruit.ABT was more suitable for the rooting of aseptic seedlings.The selection pressure of Kanamycin to epicotyls was 60 mg/L.