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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.
The hypocotyls of the sterile seedlings of C. angustifolia were used as explants to establish an in vitro regeneration system by inducing, multiplying and rooting of adventitious buds, and the contents of echinacoside and verbascoside in test-tube seedlings were determined by HPLC.The results showed that, In MS + 1.0mg / L BA + 0.5mg / L NAA medium, the induction rate of adventitious buds on hypocotyls 20d was 88.33%. In MS + 0.5mg / L BA + 0.2mg / L NAA medium, adventitious buds The average breeding coefficient of 1 ~ 3 generations reached 7.5; in 1/2 MS medium, the rooting rate of adventitious buds reached 100% at 20 days; 48 test-tube seedlings were transplanted into the field and 46 survived 30 days, the survival rate was 95.83%. The whole plant contains verbascoside about 40.37μg / g DW, excluding echinacoside.This experiment successfully established the regeneration system of Eucalyptus willdiensis, which laid the foundation for rapid propagation and genetic transformation.