白花蛇舌草叶片离体培养及试管无性系的建立

In-vitro culture of Hedyotis diffusa leaves and establishment of regeneration system

以白花蛇舌草叶片为材料,建立了白花蛇舌草叶片高效不定芽发生、植株再生体系。研究了不同激素及其组合对外植体不定芽发生的效应。结果显示,在MS基本培养基中单纯添加6BA,当6BA浓度为0.1mg/L和0.5mg/L时,不能诱导离体叶片发生不定芽,6BA浓度在1.0～5.0mg/L范围内,诱导率随BA浓度升高而增加,适宜浓度为3.0mg/L;当6BA与NAA配合使用时,其诱导率随着培养基中6BA与NAA的相对比值的提高而提高;以MS+BA3mg/L+NAA0.01mg/L作为继代培养基,建立起白花蛇舌草高效、稳定的试管无性系,为白花蛇舌草遗传转化研究奠定了基础。

High frequency adventitious shoot formation and plant-let regeneration system were established through in-vitro culture of Hedyotis diffusa leaves. The results showed that leaf adventitious shoots could not be induced on basal MS medium supplemented with 6-BA 0. 1 mg/L or 6-BA 0.5 mg/L. When 6-BA 1.0-5.0 mg/L was added in MS medium, adventitious shoots generated. The higher the concentration of 6-BA was, the higher the regeneration rate and number of adventitious shoot per leaf were obtained. However, MS medium added with 6-BA 3.0 mg/L was suitable for leaf adventitious shoot formation and growth. The combination of 6-BA and NAA promoted the regeneration efficiency of leaf in-vitro. This study indicated that it was the ratio of 6-BA/NAA rather than the quantity that influenced the regeneration of adventitious shoot. The optimal ratio of 6-BA/NAA was 10. MS basal medium supplemented with BA 3 mg/L and NAA 0. 01 mg/L was used as subculture medium and a high frequency and stable regeneration system was built, which laid a foundation for Agrobacterium-mediated genetic transformation.