【作者】 李翠花；

【导师】 吴震；

【作者基本信息】 南京农业大学 ， 蔬菜学， 2007， 硕士

【摘要】 宜兴百合别名虎皮百合、黄百合，分类学定名为卷丹[Llium.lancifolium Thunb.（Lilium.tigrinum ker.-Gawl.）]，原产太湖一带的湖州和宜兴。其食用部位是鳞茎，不仅有着丰富的营养价值，而且有着重要的药用价值，被誉为“太湖参”、“中条参”。宜兴百合以无性繁殖为主，但传统的百合繁殖方法，繁殖系数较低，并且长期的营养繁殖，常造成病毒积累和种性退化，影响百合的产量和质量。组织培养技术广泛应用于百合引种栽培，优良品种快速繁殖，去除病毒和更新品种，能够加快百合快速繁殖的速度，缩短百合的生育周期。虽然国内外学者对百合组织培养做了大量的研究，但是对宜兴百合（卷丹）的关注还不够，并且百合种间基因型差异大，在离体培养中的表现也有一定的差异。因此，系统研究宜兴百合离体再生中的各影响因子，探讨不同离体再生途径对解决宜兴百合的种苗繁殖问题是十分必要的。本试验以宜兴百合为试验材料研究了不同外植体及处理方式对离体器官发生的影响，探讨了生长调节物质和培养基对形态发生的作用，并对试管鳞茎形成进行了初步的研究。主要研究结果如下：1、珠芽是宜兴百合一种理想的外植体来源。MS+NAA0.2mg／L+6-BA1.5mg／L+30g／L蔗糖+6g／L琼脂粉培养基中，珠芽不定芽直接再生效果最好，诱导率和分化系数分别达93.8％、5.32。而以试管苗基部为外植体于MS+2，4-D1.0mg／L+6-BA0.5mg／L+1.0g／LAC+30g／L蔗糖+6g／L琼脂粉培养基上培养40d，可获得较高的诱导率和分化系数。宜兴百合的离体培养中，不仅鳞片表现出位置效应，其珠芽和叶片在培养中也存在位置效应。因此，在宜兴百合离体培养中，应以所选器官的形态学下端作为外植体，更易获得成功。2、2，4-D是诱导宜兴百合鳞茎和试管鳞茎形成愈伤组织的必需物质，并且2，4-D浓度为2.0mg／L时，外植体愈伤组织诱导率最高；细胞分裂素种类对愈伤组织形成有很大影响，KT诱导胚性愈伤组织的效果比6-BA好。宜兴百合鳞茎及试管鳞茎鳞片均可在MS+2.0mg／L2，4-D+0.5KT+6L琼脂粉+30g／L蔗糖中，诱导产生胚性愈伤，并可进一步发育成苗。百合试管鳞茎鳞片离体再生过程中，生长调节物质的种类和浓度影响其再生器官的类型，KT、GA₃均可促进试管鳞茎鳞片直接形成小鳞茎。基本培养基类型影响宜兴百合鳞茎直接再生的器官类型，无论是培养基中KH₂PO₄加倍，还是除去NH₄NO₃加倍KNO₃，均有利于小鳞茎的直接发生。但以百合试管鳞茎鳞片为外植体，基本培养基对其再生器官的类型影响不大，以不定芽直接发生为主。此外，在百合试管鳞茎鳞片离体再生过程中，生长调节物质的种类和浓度，特别是KT、GA₃均可促进试管鳞茎鳞片直接形成小鳞茎。3、蔗糖对宜兴百合试管鳞茎的直径、鲜重都有很大的影响，90g／L蔗糖是诱导宜兴百合试管鳞茎形成和膨大的理想浓度。在一定时间范围内，适当延长百合继代周期，可以促进植株营养物质向试管鳞茎转移，进而促进试管鳞茎膨大。植物生长抑制剂PP₃₃₃在对植物茎叶生长产生抑制作用的同时可促进试管鳞茎的形成和膨大。以5.0mg／L诱导效果最佳。SA对植株叶片和根的生长产生明显的抑制作用，同时也有效的增大鳞茎的直径，形成鳞茎的整齐度增加，其中以0.1mg／L SA效果最好。但是添加SA与对照相比，鳞茎鲜重有所下降。因此，如何采取一定保护措施，保持SA对百合试管鳞茎形成有利影响的同时，提高试管鳞茎重量，有待进一步研究探讨。更多还原

【Abstract】 Yixing lily, also called ’tiger lily’ or ’yellow lily’, is denominated Lilium lancifolium Thunb （L. tigrinum ker.-Gawl.） according to the plant classification. Its origins are Huzhou and Yixing in Tai Lake area, Jiangsu province. The edible parts of Yixing lily are bulblets which not only has plentiful nutrition but also important medicinal value. Yixing lily is know as ’Tai Lake Ginseng’ or ’Zhongtiao Ginseng’. Yixing lily mainly depends on asexual propagation, i.e. vegetative propagation. Lower propagation coefficient, accumulation of viruses and varietal degeneration after long periods of vegetative propagation in the traditional reproduction method reduce the yield and quality of lily. Recently, tissue culture has been extensively applied in the introduction and cultivation, virus removal, variety regeneration, and rapid propagation of fine varieties of lily, expediting the propagation rate and shortening the growth period.Although there has a lot of work been done on lily, little attention has been paid to theYixing lily. Since the genotype differences among varieties of lily, differences also exist in tissue culture. It is very important to study on the factors related to the regeneration of Yixing lily in vitro and to discuss different methods in solving the problems in the production.In this study,Yixing Lily was used as explant to study effects of different explant and different treatment on in vitro regeneration pathways of Yixing Lily,meanwhile preliminary study on bulblet formation were curried out.The main results were as follow.1.Bulblet is an ideal source of explant.the medium contained MS+NAA0.2mg/L+6-BA1.5mg/L+30g/Lsucrose+6g/Lagar is more effective in inducing adventitious buds from bulblets,with inductivity 98.3% and differentiation coefficient 5.32. While 2,4-D1.0mg/L combined with 6-BA0.5mg/L,plantlet’s leaf can obtain high inductivity and differentiation coefficient.In vitro culture of Yixing Lily,not only bulbscale but bulblet and leaf have position effect. So the bottom part of the organ shoud be used as explant and it may success easily. 2.2,4-D is the essential plant growth regutor in inducing callus from bulbscale and bulblet of Yixing Lily.Different cytokinin has different effects on callus induction.KT has the better effect on callus induction than 6-BA.Both bulbscale and bulblet in vitro of Yixing Lily can be inducted to form embryogenic callus in the medium contained.Plant growth regulator especially KT and GA₃ may also influent the type of organ regenerated directly from bulblet in vitro,which can improve bulblet regeneration.Furthermore,Basic media influent organogenesis from bulbscale,not only to twice KH₂PO₄ but to remove NH₄NO₃ and twice KNO₃ can improve bulblet to regenerate directly.But basic media has less influence on the type of organs regenerated by organogenesis from bulblet in vitro,and adventitious buds regenerated directly in every test.3. Sucrose had great effect on the diameter,fresh mass of bulblet,90g/L was the ideal concentration in inducing and swelling bulblet from plantlet of Yixing Lily.Extending subculture cycle appropriately could improve nutritive material transferred to bulblet,which could improve bulblet swelling.Plant retardant PP₃₃₃ can restrain plant growth and at the same time it can promote bulblet forming and swelling.The best effect was 5mg/L PP₃₃₃.SA can retain the growth leaves and roots obviously.and it can increase the bulblet formation rate, the diameter and uniformity of bulblet. The best effect was 0.1 mg/LSA.But bulblet formed in the media with SA have less fresh mass than the CK.So it is a moot question how to take some protect measures to maintain the advantage of SA and enhance bulblet fresh mass.更多还原