卵叶钓钟柳的离体再生及特殊中药成分的HPLC检测

Regeneration in vitro of Penstemon ovatus Plantlets and Determination of Their Special Chinese Medicinal Ingredients by HPLC

以卵叶钓钟柳无菌苗的下胚轴为外植体,通过不定芽诱导、增殖和生根建立离体再生体系,并采用HPLC检测试管苗中松果菊苷和毛蕊花糖苷含量.结果表明,在MS＋1.0mg/L BA＋0.5mg/L NAA培养基中,下胚轴20d不定芽诱导率为88.33%;在MS＋0.5mg/L BA＋0.2mg/L NAA培养基中,不定芽1~3代平均繁殖系数达7.5;在1/2 MS培养基中,不定芽20d生根率达100%;48株试管苗移栽到田间,30d成活46株,成活率达95.83%.试管苗全株含毛蕊花糖苷约40.37μg/g DW,不含松果菊苷.该试验成功建立了卵叶钓钟柳离体再生体系,为其快速繁殖和遗传转化奠定了基础.

Hypocotyls from germ‐free Penstemon ovatus plantlets were used as explants to establish a plant‐let regeneration system in v itro of P. ov atus via adventitious bud induction , proliferation and rooting , HPLC was employed to determine the contents of echinacoside and verbascoside in the plantlets .The results showed that the maximum regeneration frequency （88.33% ） of adventitious buds was achieved after the hypocotyls were cultured on MS＋1.0 mg/L BA＋0.5 mg/L NAA for 20 days;that on MS＋0.5 mg/L BA＋0.2 mg/L NAA the average multiplication coefficient of the adventitious buds was as high as 7.5 from 1 to 3 subcultures ;and that 100% of the adventitious buds rooted readily on 1/2 M S medium for 20 days .Forty‐eight plantlets w ere transplanted to a matrix soil and 46 survived after 30 days , the survival rate being 95.83% .The plantlets contained verbascoside by about 40.37 μg/g DW ,and no echinacoside was detected by HPLC .The plant regeneration system in vitro of P. ovatus is thus successfully estab‐lished .This procedure of the culture method may be very useful for rapid propagation of P. ovatus and for introduction of foreign genes into P. ov atus .