牡丹和芍药组织培养的研究(1)

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1、河南科技大学 硕士学位论文 牡丹和芍药组织培养的研究 姓名：李海刚 申请学位级别：硕士 专业：植物学 指导教师：孔祥生 20100501 摘 要 I 论文题目：论文题目： 牡丹和芍药组织培养的研究牡丹和芍药组织培养的研究 专专 业：业： 植植 物物 学学 研研 究究 生：生： 李李 海海 刚刚 指导教师：指导教师： 孔孔 祥祥 生生 教教 授授 摘 要摘 要 本试验以牡丹和芍药为材料，主要研究了牡丹的胚培养、芽培养和影响试管 苗玻璃化的因素，并且研究了芍药的休眠芽培养、愈伤组织的诱导和褐化的防 止，结果如下： 1牡丹胚培养 以凤丹成熟胚为外植体，研究发现不同的培养基对牡丹胚萌发率有明显 的影响

2、，MS 可以作为凤丹胚培养的基本培养基，凤丹胚在培养基 MS+6-BA 1.0 mg/L+GA3 1.0 mg/L 上的萌发率最高，MS+6-BA 1.0mg/L+KT 1.0 mg/L 是适合试管苗继代培养的培养基，试管苗在培养基 1/2MS+IBA 2.0 mg/L 上 的生根率达到 43.2%，牡丹种子经过 45d 的低温处理，胚萌发率有明显的提高。 2牡丹芽培养 以牡丹休眠芽为材料，采用 0.1% HgCl2 10min 进行消毒既可减少污染又能 减少试管苗的玻璃化。不同浓度的 6-BA 对不同牡丹品种休眠芽萌发率的影响不 同。利用正交设计对洛阳红试管苗增殖试验进行了研究，结果表明 6

3、-BA 和 KT 是洛阳红试管苗增殖过程中重要的生长调节物质，洛阳红试管苗增 殖的最优组合为 MS+6-BA 1.0 mg/L+KT 1.0 mg/L，增殖系数为 2.15。不同品种 在相同培养基上的增殖系数有明显差异。洛阳红试管苗在培养基 1/2MS+IBA 1.0 mg/L+蔗糖 30 g/L+琼脂 6 g/L 中的生根状况较好，生根率可以达 到 82.5%，两步生根法可以提高试管苗的生根率。 3牡丹试管苗玻璃化的防止 以 MS 为基本培养基，研究牡丹品种、琼脂、6-BA、大量元素和蔗糖浓 度、光照等因素对试管苗玻璃化的影响表明，不同品种的牡丹试管苗在组织培养 过程中出现不同程度的玻璃化，

4、增加培养基中琼脂浓度、降低 6-BA 和大量元素 的浓度、增加光照强度或采用自然光照均可以有效地降低牡丹试管苗玻璃化的发 生。 4芍药休眠芽培养 以芍药休眠芽为材料，2 月份是芍药休眠芽采集的最佳时期。不同品种和不 II 同培养基对芍药休眠芽萌芽率的影响有明显的不同，1/2MS 比较适合作为诱导芍 药休眠芽萌发的基本培养基。种生粉试管苗增殖的最优组合为 MS+6-BA 1.0 mg/L+KT 0.5 mg/L，增殖系数为 2.39。不同浓度的 IBA 对试管苗的生根率、根 长和生根数都有一定的影响，芍药试管苗在培养基 1/2MS+IBA 2.0 mg/L 中的生 根效果最好，生根率可以达到 5

5、8.0%。 5芍药愈伤组织的诱导和愈伤组织褐化的防止 以种生粉为试验材料，比较不同外植体，不同基本培养基类型，不同激 素组合对愈伤诱导率的影响表明，以试管苗叶柄为外植体，在MS+6-BA 1.0 mg/L2,4-D 0.5 mg/L+蔗糖30g/L+琼脂6 g/L的培养基中诱导率最高可达80.0； 抑制褐化的研究表明，1/2WPM为最佳基本培养基，AgNO3的防止褐化效果最 好；黑暗和低温都能不同程度的降低褐化的程度。 关关 键键 词：词：牡丹，芍药，试管苗，愈伤组织，玻璃化，褐化 论文类型：论文类型：应用基础研究 摘 要 III Subject: The research on tissue

6、 culture of Paeonia suffruticosa and Paeonia lactiflora Pall Specialty: Botany Name: Li Hai gang Supervisor: Prof. Kong Xiang sheng ABSTRACT This test was based on Paeonia suffruticosa and Paeonia lactiflora Pall for materials. Paeonia suffruticosa was mainly studied on embryo culture, shoot culture

7、 and factors that affected the vitrification, and Paeonia lactiflora Pall was studied on dormant shoot culture, callus induction and prevention of browning. The results were as follows: 1. Embryo culture of peony By using Feng Dan mature embryos as explants, a procedure for rapid propagation of matu

8、re embryo culture was established. The results of experiments showed that the effect of different media on peony bud induction was significant, MS can be used as Feng Dan basic embryo culture medium, the germination rate of Feng Dan embryos in medium MS+6-BA 1.0 mg/L+GA3 1.0 mg/L was best, the mediu

9、m MS+6-BA 1.0mg/L+KT 1.0mg/L was suitable for subculture medium, the rooting rate of seedlings in the medium 1/2MS+IBA 2.0 mg/L was up to 43.2%,the induction of germination rate increased significantly after 45d of cold treatment. 2. Dormant bud culture of peony By using dormant buds of Paeonia suff

10、ruticosa as explants, Disinfection with 0.1% HgCl2 10min was better to reduce pollution and vitrification. The effect of different concentrations of 6-BA on germination rate of dormant buds of different varieties was different. Orthogonal design of the Luo Yang Hong in vitro multiplication was studi

11、ed, the results showed that 6-BA and KT were important growth regulators of Luo Yang Hong in vitro multiplication, the optimal combination of Luo Yang Hong in vitro multiplication was MS+6-BA 1.0 mg/L+KT 1.0 mg/L, multiplication coefficient was 2.15.The multiplication coefficient of different variet

12、ies on same medium was significantly different. Luo Yang Hong plantlets rooted well in the medium 1/2MS+ IBA 1.0 mg/L+sucrose 30g/L + agar 6g/L, rooting rate can reach 82.5%, rooting rate can be improved by two steps rooting method. 摘 要 IV 3. Prevention of vitrification of peony plantlet MS medium w

13、as chosen as the basic medium and the main factors such as cultivars, agar, 6-BA, macroelement, sucrose and intensity of illumination were investigated on vitrification of plantlets from Paeonia suffruticosa. The results showed that different cultivars of Paeonia suffruticosa in tissue culture proce

14、ss appeared vitrification in degree. Increasing the concentration of agar in culture medium, reducing the concentration of 6-BA, reducing the concentration of macroelement and the increasing intensity of illumination or using the natural light were possible to reduce the vitrification of plantlets e

15、ffectively. 4. Dormant bud culture of herbaceous peony By using dormant buds of Paeonia lactiflora Pall, acquisition time of dormant bud affected the germination rate to certain extent; February was the best time of the herbaceous peony dormant buds acquisition. Effects of different medium on germination rate of dormant buds of different varieties were significantly different; 1/2MS was quite suitable basic culture medium for induction of germination herbaceous peony dormant bud. the optimal combination of Zhong Sheng Fen in vitr