瓦布贝母组织培养及生物碱的测定

为缓解川贝母的市场供需矛盾,本研究利用组织培养技术生产瓦布贝母再生鳞茎,探究了适宜的组织培养条件,并采取酸性染料比色法测定总生物碱.结果发现,鳞茎采用75%酒精约10 s和0.1%升汞15 min灭菌,外植体污染率最低;不同种类和浓度范围的激素能够影响外植体的启动和生长状况,NAA和6-BA浓度范围分别控制在0.5～2.0 mol/L,均能诱导出再生鳞茎;不同激素培养条件下,瓦布贝母再生鳞茎总生物碱存在差异,有超过一半的再生鳞茎总生物碱含量达到中华药典的规定要求,因此,组织培养是获取瓦布贝母活性成分的一种有效途径.

Tissue culture and alkaloids determination of Fritillaria unibracteata Hsiao et K.C.Hsia var. wabuensis

In order to alleviate the contradiction between market supply and demand of Fritillaria cirrhosa, the regenerated bulbs of Fritillaria unibracteata Hsiao et K. C. Hsia var. wabuensis were produced by tissue culture technology in this study, the suitable tissue culture conditions were also explored, and the total alkaloids were determined by acid dye colorimetry. The results showed that the bulbs were sterilized with 75 % ethanol for about 10 s and 0.1 % HgCl2 for 15 min with the lowest explants contamination rate. Different types and concentrations hormones can affect the initiation and growth of explants, the concenteation range of NAA and 6-BA were controlled at 0.5~2.0 mol/L, respectively, and both can promote the growth of regenerated bulbs. The contents of total alkaloids in regenerated bulbs of Fritillariaunibracteata Hsiao et K. C. Hsia var. wabuensis were determined. Under different hormone culture conditions, there were differences in the total alkaloids of regenerated bulbs of Fritillaria unibracteata Hsiao et K. C. Hsia var. wabuensis, and the total alkaloids of more than half of the regenerated bulbs in tissue culture met the requirements of Chinese Pharmacopoeia. Therefore, tissue culture is an effective way to obtain the active ingredients of Fritillaria unibracteata Hsiao et K. C. Hsia var. wabuensis.Fritillaria unibracteata Hsiao et K. C.