摘要：小蔓长春花具有很高药用价值，其所含成分长春胺对缺血性心血管疾病的治疗尤其有效，被广泛应用于医药领域。但随着其用途的扩展，其资源面临着严峻局势。植物细胞培养则是解决资源短缺问题的理想途径之一。本研究选取小蔓长春花嫩叶和茎段为材料，对小蔓长春花愈伤组织诱导和细胞悬浮培养进行研究。研究结果表明，取小蔓长春花无菌苗上的茎段，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%。取出愈伤组织进行继代培养后移至MS+2.0 mg•L-12,4-D+0.2 mg•L-1 6-BA的培养基上进行细胞悬浮培养，30天后可得稳定的细胞悬浮体系。所建立的悬浮细胞系呈“S”型曲线，培养12d达到最大生长量，培养中，pH变化趋势为先下降后上升，细胞数量则呈现出先上升后下降的变化趋势。33044
毕业论文关键词： 小蔓长春花；愈伤组织；细胞悬浮培养
Study on the Technique of Cell Culture of Vinca minor L.
Abstract: The Vinca minor L. is very valuable in medical field and its active ingredient vincamine is effective on the treatment of ischemic cardiovascular disease, and it is widely used in medicine field. However, with its demand expansion, the resources are facing severe situation. Plant cell culture is one of the ideal way to solve the problem of resource shortage.This study aims to select the leaves and stem segments as materials for callus induction and cell suspension culture. The results shows that the segments of the young plants were inoculated with the MS culture with 2.0mg L-12, 4-D+0.2 mg L-1 6-BA, and the induction rate was 80%. After subculture, the cell suspension was cultured on the medium of MS of 2.0mg L-12, 4-D+0.2 mg L-1 6-BA +2.0, and we can get the stable suspension system after 30 days. The establishment of suspension cell curve is represented as "s" curve and cultured for 12 days to reach the maximum amount of growth. The value of pH decreased at first and then increased. The number of cell increased at first and then decreased.
Keywords: Vinca minor L.; Callus; Cell Suspension Culture
目录
1 前言 1
1.1 小蔓长春花的研究现状 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 植物细胞悬浮培养研究进展 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 小蔓长春花细胞培养的研究进展 9
1.4 本论文的研究内容 9
2 实验材料与方法 11
2.1 实验材料 11
2.1.1 材料 11
2.1.2 主要试剂 11
2.1.3 主要仪器 11
2.2 实验方法 12
2.2.1 预备试验 12
2.2.2 技术路线 12
2.2.3 准备试验 12
2.2.4 不同激素配比对小蔓长春花无菌苗培养的影响试验 13
2.2.5 愈伤组织的诱导 13
2.2.6 细胞悬浮培养 15
3 结果与分析 17
3.1 不同激素配比对小蔓长春花无菌苗获得的影响 17
3.2 愈伤组织诱导 18
3.2.1 不同外植体对愈伤组织诱导的影响 18
3.2.2 不同激素的配比对小蔓长春花叶片愈伤组织诱导的影响 18
3.2.3 光照对诱导小蔓长春花愈伤组织的影响 21
3.3 细胞悬浮培养 21
3.3.1 不同激素配比对细胞悬浮培养的影响 21 小蔓长春花细胞悬浮培养的研究:http://www.youerw.com/shengwu/lunwen_29902.html