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诺丽叶片的离体再生

2017-05-30黄奥丹蓝增全吴田

广西植物 2017年6期

黄奥丹 蓝增全 吴田

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·L1 6BA + 2.0 mg·L1 2,4D;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·L16BA+ 0.4 mg·L1 NAA或MS +2.0 mg·L16BA+ 0.4 mg·L1 NAA,其中MS + 1.0 mg·L16BA + 0.4 mg·L1 NAA生根時间最早为10 d左右,根系较发达,而MS + 2.0 mg·L16BA + 0.4 mg·L1 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·L16BA + 0.4 mg·L1 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·L1 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。

关键词: 诺丽, 植物激素, 愈伤组织, 再生培养

中图分类号: Q943.1

文献标识码: A

文章编号: 10003142(2017)06074908

Abstract: 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·L1 6BA + 2.0 mg·L1 2,4D; the optimal medium that induced leaf callus to generate adventitious roots and buds was MS + 1.0 mg·L1 6BA+ 0.4 mg·L1 NAA or MS + 2.0 mg·L1 6BA + 0.4 mg·L1 NAA, Thereinto, the solution of MS + 1.0 mg·L1 6BA + 0.4mg·L1 NAA made the rooting time earler, about 10 d, and its root system was more developed. Instead, the solution of MS + 2.0 mg·L1 6BA + 0.4 mg·L1 NAA made it 15 d. The optimal medium that induced leaves to generate roots and buds in mode Ⅱ was MS + 1.0 mg·L1 6BA + 0.4 mg·L1 NAA。Cutting and transplanting the seedlings regenerated in vitro from model Ⅰ and Model Ⅱ into MS + 0.2 mg·L1 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.

Key words: noni, plant hormones, callus, regeneration of culture

诺丽(Morinda citrifolia)又称海滨木巴戟、海巴戟天、檄树(吴田和蓝增全,2011;曹艳花,2014),为茜草科(Rubiales)巴戟天属(Morinda)植物(杨小波2013),是一种热带常绿多年生阔叶灌木或小乔木。主要生长于温暖潮湿的热带、亚热带地区,在国外主要分布于南太平洋诸岛,如大溪地、夏威夷、印度尼西亚等,在我国主要生长于海南岛、西沙群岛、云南西双版纳及台湾南部等地区(何明霞和杨清,2006)。现代医学研究证明, 诺丽果汁中具有免疫调节及抗肿瘤作用的多糖成分(张学梅,2000), 果实单用或与其他植物合用可治疗多种疾病,被医学研究者称为当代神奇的治疗物质(赖茂良,2014)、(段文荣等,2009),有较高的经济价值。……

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