金银花组织培养研究.docxVIP

摘要： 为了加快金银花产业的发展。提高金银花的繁殖系数，避免生态环境被破坏。以药用 金银花的当年生幼叶为外植体，用MS基本培养基。通过不同的生长激素和细胞分裂素浓 度梯度，筛选适合的培养浓度和培养条件。实验结果表明，随着培养基6-BA浓度的升 高，褐变率随Z提高，褐变现象出现的时间也越早；反Z,较低浓度的6-BA适宜幼叶的 分化生长，褐变反应慢。另外，培养时间过长，未及时转接，也出现了褐变现象。采用3 X3 （6?BA,NAA各3个平行组）正交实验的方法,找出最优水平组合，得出最佳的激素 配比纽合。培养条件：26°C恒温培养，光照12001xo实验结果表明，诱导愈伤组织的增殖 最佳培养基：MS+6-BA0.5mg/L+NAA0.5 mg/L;诱导芽分化的最佳培养基：MS+6-BA1.0 mg/L+NAAO」mg/L。 关键词：金银花；外植体；生长激素；细胞分裂素 Abstract To quicken the development of Honeysuckle industry, improve the proliferation rate of Honeysuckle, avoid the damage of ecological environment, we choose current-year-young leaves of Jiangsu medicinal Honeysuckle to establish aseptic system for explants.In the whole cultivation process, we choose MS medium. The disinfection method: 1.Sterilization with alcohol of 75% concentration for 45s, 2.Sterilization with HgC12 of 0.15% concentration for 8niin, 3.Washing with aseptic water for 4 or 5 times, 4.Drying with aseptic filter pape匚There two kinds of medium: with activated carbon and without activated carbon. Experimental results demonstrated that a little of activated carbon can decrease Browning. With comparison of different cultivation time and concentration of growth hormone(NAA) and cytokinin(6-BA), we discover that the higher of the 6-BA concentration, the higher Browning rate and the earlier of Browning. On the contrary, young leaves growth better and later Browning with lower concentration of 6-BA? We also found Browning with too long cultivation time. By 3 X3 Orthogonal Experiment, We find optimal combination of hormone. Cultivation environment: constant temperature with 26 °C, light irradiaton 12001x. The main results: LThe best multiplication medium for callus: MS+6-BA0.5mol/L+NAA0.5 mol/L.2 The best medium for meristtem culture: MS+6-BA1.0 mol/L+NAA0.1 mol/L. Key words： Honeysuckle, aseptic system, growth hormone, cytokinin 1前言 1.1形态特征及生物学特征 金银花（拉丁学名：Lonicera Japonica,英文名：Japanese Honeysuckle）别称：忍冬、 金银藤、银藤、二色花藤、二宝藤、右转藤、了风藤、鸳鸯藤。属于植物界，被了植物门 Angiospermso 合瓣花亚纲 core eudicot