三叶青细胞悬浮培养体系的条件优化和总黄酮含量的测定研究  

Study on condition optimization of Cymbidium trifolium cell suspension culture system and determination of total flavonoids content

作　　者：何海燕 付跃[1,2,3] 覃拥灵 张健 HE Hai-yan;FU Yue;QIN Yong-ling;ZHANG Jian

机构地区：[1]河池学院化学与生物工程学院,广西河池546300 [2]微生物及植物资源开发利用广西高校重点实验室,广西河池546300 [3]河池学院农业生物技术应用研究中心,广西河池546300 [4]广西卫生职业技术学院,广西南宁530023

出　　处：《饲料研究》2023年第7期77-80,共4页Feed Research

基　　金：广西自然科学基金(项目编号:2020GXNSFAA297218)。

摘　　要：试验旨在探讨使用三叶青细胞悬浮培养生产三叶青黄酮的可行性。试验以野生三叶青叶片和茎段为外植体诱导愈伤组织,测定增殖、单细胞培养及其总黄酮含量,悬浮培养细胞经不同液体培养基配方、光照方式、摇床转速、接种量、pH值、培养天数培养,以乙醇为溶剂,回流提取总黄酮,使用NaNO_(2)-Al(NO_(3))_(3)显色,在500 nm处用紫外分光光度计测定总黄酮含量。结果显示,三叶青最佳培养基为MS+1.5 mg/L 6-BA+2.5 mg/L 2-4D+3 mg/L NAA+0.4 mg/L 6-KT,8 h/d光照,摇床转速110 r/min,接种量25 m L/L,pH值5.8,培养时间为13 d。研究表明,三叶青细胞培养可生产黄酮,最优培养条件下三叶青细胞悬浮培养体系黄酮积累量可达20.524 mg/g。The experiment was to investigate the feasibility of producing trifolium flavone from Cymbidium trifolium cell suspension culture.The callus was induced by leaves and stems of wild Cymbidium trifolium as explants,and the proliferation,single cell culture and total flavonoid content were measured.The suspension cultured cells were cultured with different liquid medium formulations,light mode,shaking table speed,inoculum amount,pH value and culture days.The total flavonoids were extracted by reflux with ethanol as solvent,and NaNO_(2)-Al(NO_(3))_(3) was used for color development.Total flavonoid content was determined by UV spectrophotometer at 500 nm.The results show that the optimal medium for Cymbidium trifolium was MS+1.5 mg/L 6-BA+2.5 mg/L 2-4D+3 mg/L NAA+0.4 mg/L 6-KT,8 h/d illumination,shaking speed 110 r/min,inoculation dosage 25 mL/L,pH value 5.8,incubation time was 13 d.The results show that flavonoids could be produced by Cymbidium trifolium cell culture,and the accumulation of flavonoids in the suspension culture system of Cymbidium trifolium cell could reach 20.524 mg/g under optimal culture conditions.

关 键 词：三叶青 细胞培养 总黄酮 

分 类 号：S816.7[农业科学—饲料科学]