枇杷细胞悬浮培养生产熊果酸的调控

Regulation of Ursolic Acid Content in Suspension Culture of Eriobotrya japonica L.

以枇杷悬浮培养细胞的生长量及熊果酸(UA)的含量为考察指标,从蔗糖浓度、装液量、p H值、接种量、温度、光照强度、摇床转速、激素等开展培养参数的筛选。结果表明:接种量8 g/100 m L,p H6.0,蔗糖30 g/L,25℃,白炽灯400 lx,转速130 r/min,装液量130 m L/250 m L,MS+NAA 0.5 mg/L,15 d收获,细胞鲜重是起始的3.577倍,UA含量为34.993 mg/g DW,是自然界枇杷叶的约3.5倍。

Ursolic acid （UA） is an activate component found in loqaat and it has high economic value. The traditional UA extraction methods has limitations. A novel approach of producing UA using loquat cell suspension culture technique would have a significant benefit theoretically and practically. Different culture conditions on cell growth and UA accumulation in loquat cells, including sucrose concentration, plant growth regulators, pH, media volume, inoeulums quantity, temperature, light intensity and shaking speed were investigated. The results showed that the weight of fresh cell were 3.577 times under optimal protocol after harvesting for 20 days when compared with the initial inoeulums, and the UA content obtained reached 34.993 mg/g DW （Dry weight）. The optimal protocol was executed by transferring 8 g/100 mL of initial inoculums to a 130 mL liquid medium containing 30 g/L of carbon source, 0.5 mg/L plant hormone 1-Naphthaleneacetic acid （NAA） with pH6.0. Then loquat cell suspension was kept in a 25 ℃ shaker with 400 lx light intensity and shaking at 130 r/min for 15 days. The reulsts would provide foundations for future industrialized extracting UA from loquats in a large scale.