摘要
目的:建立以七叶树种胚为外植体进行组织培养生产七叶皂苷的体系。方法:采用正交试验法探讨了超声波法提取七叶树种胚愈伤组织中七叶皂苷的最佳工艺;通过改变激素类型和浓度,筛选出适合七叶树种胚愈伤组织增殖的最佳培养基,并对愈伤组织细胞生长与次生代谢物积累的规律进行初步分析;利用水杨酸Fenton试剂法和邻苯三酚自氧化法对愈伤组织提取液的抗氧化性进行初步探索。结果:七叶树种胚愈伤组织中七叶皂苷的最佳提取工艺为:超声温度30℃,超声时间40 min,乙醇浓度60%,液料比70∶1。愈伤组织增殖培养的最佳培养基为MS+2,4-D 1.0 mg/L+6-BA 8.0 mg/L+NAA 0.5 mg/L,在最佳增殖培养基上,愈伤组织的最佳继代和收获期均为25 d。对愈伤组织比生长速率与七叶皂苷比合成速率进行分析,发现七叶树种胚愈伤组织细胞生长与次生代谢产物的合成没有一定的规律性。愈伤组织提取液对超氧自由基和羟自由基都具有一定的清除作用。结论:该研究建立了七叶树种胚细胞培养生产七叶皂苷的体系。
Objective： To establish the system of producing escin by tissue culture of Aesculus chinensis embryos.Methods： Study on the best extraction process of escin in callus of Aesculus chinensis embryos by ultrasonic method using the orthogonal test.The optimal media for callus proliferation and high content of escin were filtered and the growth of callus and accumulation of secondary metabolites were preliminarily analyzed.The antioxidant activity of escin extract from callus of Aesculus chinensis embryos were studied by salicylic acid reagent and pyrogallol autoxidation method.Results： The optimum conditions were as followed： ultrasonic temperature 30 ℃,ultrasonic time was 40 min,ethanol concentrations was 60% and liquid-solid ratio was 70∶ 1.The optimal medium for callus proliferation of embryos in Aesculus chinensis embryos was MS ＋ 2,4-D 1.0 mg/L ＋ 6-BA 8.0 mg/L ＋ NAA 0.5 mg/L.In the optimal medium,the best subculture period and the best harvest time were 25 d.The callus specific growth rate and escin specific synthesis rate were analyzed which indicated that the growth of callus cell and the synthesis of secondary metabolites had no consistency.The extract of callus had a certain scavenging effect on superoxide radical and hydroxyl radical.Conclusion： Aesculus chinensis embryo cell culture system is established to produce escin in this study.
出处
《中药材》
CAS
北大核心
2017年第7期1552-1557,共6页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(31470668)
关键词
七叶树种胚
愈伤组织
七叶皂苷
提取工艺
激素配比
抗氧化性
Aesculus chinensis Bge. embryos
Callus
Escin
Extraction process
Hormone proportion
Antioxidant activity
