诺丽叶片的离体再生

摘要

以诺丽(Morinda citrifolia)叶片为外植体,在添加不同激素种类和浓度的MS培养基上进行离体培养,建立两种离体再生模式:模式Ⅰ为先脱分化愈伤组织,再分化不定根和不定芽;模式Ⅱ为直接培养生根后分化不定芽.结果表明:在模式Ⅰ中,诱导诺丽叶片产生愈伤组织的最优培养基为MS + 0.1 mg·L-1 6-BA + 2.0 mg·L-1 2,4-D;诱导叶片愈伤组织再分化出不定根和不定芽的最优培养基为MS +1.0 mg·L-16-BA+ 0.4 mg·L-1 NAA或MS +2.0 mg·L-16-BA+ 0.4 mg·L-1 NAA,其中MS + 1.0 mg·L-16-BA + 0.4 mg·L-1 NAA生根时间最早为10 d左右,根系较发达,而MS + 2.0 mg·L-16-BA + 0.4 mg·L-1 NAA生根时间在15 d左右,根系发达.在模式Ⅱ中,诱导叶片直接生根长芽的培养基为MS + 1.0 mg·L-16-BA + 0.4 mg·L-1 NAA.将模式Ⅰ和模式Ⅱ中,完成诺丽叶片离体再生的苗切下后接种到MS + 0.2 mg·L-1 NAA培养基中诱导生根,15 d左右分化出不定根,45 d获得完整植株.该研究结果为后续的遗传转化和基因改良研究奠定了基础.%Using noni (Morinda citrifolia) leaves as explant for culture in vitro on 3% MS medium with different types and concentrations of plant hormones,two kinds of in vitro regeneration modes were built.Mode Ⅰ: the callus was induced first,and then the adventitious roots and buds were induced;Mode Ⅱ: the adventitious roots were induced,and then the buds were induced directly.The results showed that the optimal medium of generating callus by noni leaves in Mode I was MS + 0.1 mg·L-1 6-BA + 2.0 mg·L-1 2,4-D;the optimal medium that induced leaf callus to generate adventitious roots and buds was MS + 1.0 mg·L-1 6-BA+ 0.4 mg·L-1 NAA or MS + 2.0 mg·L-1 6-BA + 0.4 mg·L-1 NAA,Thereinto,the solution of MS + 1.0 mg·L-1 6-BA + 0.4mg·L-1 NAA made the rooting time earler,about 10 d,and its root system was more developed.Instead,the solution of MS + 2.0 mg·L-1 6-BA + 0.4 mg·L-1 NAA made it 15 d.The optimal medium that induced leaves to generate roots and buds in mode Ⅱ was MS + 1.0 mg·L-1 6-BA + 0.4 mg·L-1 NAA.Cutting and transplanting the seedlings regenerated in vitro from model Ⅰ and Model Ⅱ into MS + 0.2 mg·L-1 NAA medium to induce rooting.Getting the differentiation of adventitious root about 15 d and complete plant 45 d.This study provides useful references for the breeding and the application of genetic transformation technique in noni.