大花金挖耳细胞悬浮系的建立及黄酮类化合物的产生

Optimization of Cell Suspension Culture of Carpesium macrocephalum for Flavonoid Production

研究不同培养基、蔗糖质量浓度、pH值、接种量、NAA、6-BA、VB1及培养时间对大花金挖耳细胞生长和黄酮类化合物合成的影响。结果表明:NT液体培养基在pH5.5、蔗糖质量浓度40g/L、接种量40g/L鲜质量细胞、NAA1.0mg/L+6-BA0.2mg/L时有利于细胞的生长和黄酮类化合物合成,向NT培养基中添加1.0～4.0mg/LVB1有抑制细胞褐化的作用,大花金挖耳悬浮细胞的生长及黄酮类化合物合成随培养时间的延长,表现出先升高后降低的趋势,在接种15～21d后,细胞生物量可达23.54～24.51g/L,黄酮类化合物得率为1.19%～1.23%。

The goal of the present study was to optimize the culture conditions for the establishment of suspension cell lines from Carpesium macrocephalum callus.The effects of medium type,concentrations of sucrose,VB1,NAA and 6-BA,initial pH value,inoculum quantity and culture time on cell biomass and flavonoid production were examined.The optimal conditions for culture in NT liquid medium conducive to both cell growth and flavonoid production were as follows：initial pH,5.5,inoculum quantity,40 g/L;NAA concentration,1.0 mg/L;and 6-BA concentration,0.2 mg/L.Cell browning was inhibited by adding 1.0-4.0 mg/L of VB1.Both the growth of Carpesium macrocephalum cells and the synthesis of flavonoids trended to first ascend and then descent with prolonged culture time.A cell biomass ranging from 23.54 to 24.51 g/L was achieved at 15-20 d post-inoculation,and the yield of flavonoids was between 1.19% and 1.23%.