[关键词]
[摘要]
目的 研究由石蒜愈伤组织建立细胞悬浮培养体系的条件及其生物碱的累积。方法 以石蒜鳞茎为外植体进行愈伤组织的诱导,筛选出优质的愈伤组织进行悬浮培养,考察了激素、接种量、肌醇和条件培养对石蒜悬浮细胞系建立的影响,并利用HPLC检测石蒜悬浮细胞和培养基中生物碱的量。结果 石蒜悬浮细胞系培养的最佳条件为:MS为基本液体培养基,添加2, 4-D 0.5 mg/L+KT 0.1 mg/L,肌醇300 mg/L,接种量15%;对数期的培养基能明显缩短细胞系生长的延迟期;细胞系和培养基中加兰他敏分别是石蒜鳞茎中的4.18和0.69倍。结论 利用石蒜愈伤组织建立了较好的细胞悬浮培养体系,且石蒜悬浮细胞系是一种获取加兰他敏的较好途径。
[Key word]
[Abstract]
Objective To investigate the culture conditions for the establishment of cell suspension culture system for Lycoris radiata callus and its alkaloid accumulation. Methods Using the bulbs of L. radiata as explants for callus induction, calli with good quality were selected for suspension cultivation. The factors influencing the establishment of L. radiata cell suspension culture system, such as hormone, inoculum amount, inositol, and conditional culture, were investigated. The alkaloid content in cell suspension culture system of L. radiata was determined by HPLC. Results The optimal medium for the cell suspension culture was MS + 0.5 mg/L 2, 4-D + 0.1 mg/L KT + 300 mg/L inositol and the inoculum size was 15%. The lag phase of the cell suspension culture system could be evidently shortened by the medium of logarithmic growth. The galanthamine contents in the cell suspension culture system and the medium were 4.18 and 0.69 times of those in the bulbs of L. radiata, respectively. Conclusion The cell suspension culture system of L. radiata is established and it is a feasible way to gain galanthamine.
[中图分类号]
[基金项目]
安徽省自然科学基金面上项目(1208085mc48);国家科技支撑计划项目(2013BAJ10B12)