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摘要：本研究旨在选出适宜外植体进行莪术的离体培养研究，以建立植株再生体系。研究中，主要选用莪术的根茎上的嫩芽或嫩叶作为外植体，接种于基本培养基，外加不同浓度激素组合进行无菌苗的培养、愈伤组织的诱导、丛生芽增殖和分化培养的研究，找出较为适合莪术组织培养的培养基配方，旨在摸索出较优化的再生体系的条件。以期为药用植物莪术的规模化生产、开发利用，以及莪术细胞培养的深入研究奠定基础。研究结果表明，莪术嫩芽在0.1%的HgCl2溶液中消毒10min，然后接种于MS+2.0 mg•L-1 6-BA+0.2 mg•L-1 NAA的培养基中，30d后能获得较好的莪术无菌苗。随后取无菌苗上的嫩叶，接种于MS+2.0 mg•L-12,4-D+0.2 mg•L-1 6-BA的培养基中诱导愈伤组织，诱导率高达80%。取出愈伤组织移至芽分化培养基中进行不定芽的诱导，适宜MS+2.0 mg•L-16-BA+0.2 mg•L-1 的NAA的培养基中诱导。22444
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毕业论文关键词：莪术；组织培养；愈伤组织；不定芽
Study on the Technique of Tissue Culture of Curcuma Zedoaria
Abstract: In order to select a suitable explant for the research In vitro culture of Curcuma Zedoaria, the study aims at establishing the system of plant regeneration. In the study, rhizome of buds or leaves were used as explant to be seeded in medium. And adding different concentrations of the hormone to conduct aseptic seedling culture, callus induction, induce cluster buds to multiply, to find the suitable medium for culture of curcuma, and the optimized conditions for regeneration system. Lay the foundation to large-scale production, development and utilization, and In-depth study of Curcuma cell culture. Result shows：To get the aseptic seedling after 30 days, the Curcuma Zedoaria buds should be immersed in 0.1% HgCl2 for 10 minutes. Then they were inoculated in the medium of MS +2.0 mg•L-1 6-BA + 0.2 mg•L-1 NAA . Then taking some leaves from aseptic seedling, and inculating in the medium of MS +2.0 mg•L-1 2,4-d + 0.2 mg•L-1 6-BA to get the callus, the callus induction rate was 80%.Finally, moving callus to buds medium to induct the adventitious buds. The better medium for adventitious buds is MS+2.0 mg•L-16-BA and 0.2 mg•L-1 NAA.
Keywords: Curcuma Zedoaria; tissue culture; callus; adventitious buds
目录
1 前言 1
1.1 莪术的研究现状 2
1.1.1 化学成分的研究 2
1.1.2 药理学的研究 2
1.1.3 植物资源概况 4
1.2 植物组织培养研究进展 5
1.2.1 植物的快速繁殖研究进展 5
1.2.2 植物组织和器官培养生产次生代谢产物的研究概况 7
1.3 莪术组织培养的研究进展 8
1.4 本论文的研究内容 9
2 实验材料与方法 10
2.1 实验材料 10
2.1.1 材料 10
2.1.2 主要试剂 10
2.1.3 主要仪器 10
2.2 实验方法 11
2.2.1 预备试验 11
2.2.2 技术路线 11
2.2.3 准备试验 11
2.2.4 无菌苗的获得 12
2.2.5 愈伤组织的诱导 13
2.2.6 不定芽的诱导 14
3 结果与分析 16