苦豆子愈伤组织培养及其药用成分含量测定

Callus Culture of Sophora Alopecuroides L.and Its Medicinal Components Determination

本研究利用组织培养技术考察外植体种类、激素水平、水解酪蛋白浓度等不同因子对愈伤组织的生长状态的影响;同时采用分光光度法和HPLC测定不同继代次数愈伤组织中总黄酮和槐定碱的含量。结果表明,苦豆子愈伤组织诱导的最适外植体是子叶;愈伤组织诱导的最佳培养基为MS+2,4-D0.5mg/L+6-BA1.0mg/L+NAA0.5mg/L,最佳继代培养基为MS+2,4-D1.0mg/L+6-BA4.0mg/L+NAA1.0mg/L+CH1000mg/L;总黄酮和槐定碱在第5次继代培养的愈伤组织中含量最高,分别为0.53%和0.0112%。因此,可选择第5次继代培养的愈伤组织作为苦豆子细胞悬浮培养的材料。

The effects of explants type,hormone levels,casein hydrolysate on the growth of callus were measured by tissue culture.Meanwhile,the contents of total flavonoids and sophoridine in callus of different subculture times were determined by ultraviolet spectrophotometry and HPLC.The results showed that the cotyledon was optimum explants of callus induction,the best medium of callus induction was MS＋2,4-D 0.5 mg/L＋6-BA 1.0 mg/L＋NAA 0.5 mg/L,and the best subculture medium was MS＋2,4-D 1.0 mg/L＋6-BA 4.0 mg/L＋NAA 1.0 mg/L＋CH 1000 mg/L.Contents of total flavonoids and sophoridine in callus of fifth subculture had the maximum,and 0.53% and 0.0112% respectively.So,the callus of fifth subculture was selected to use for material of cell suspension culture of S.alopecuroides.