卡那霉素对紫花苜蓿愈伤组织诱导和分化的影响

Effects of Kanamycin on Induction and Differentiation of Medicago sativa L Callus

[目的]研究紫花苜蓿叶片在诱导愈伤组织阶段、愈伤组织分化阶段对卡那霉素的敏感程度。[方法]确定卡那霉素对紫花苜蓿叶片愈伤组织形成的影响,并在形成愈伤组织的基础上,进一步确定卡那霉素对愈伤组织分化的影响。[结果]不同浓度的卡那霉素对紫花苜蓿叶片诱导愈伤与分化均有一定的抑制作用,而且随浓度升高,抑制作用增强;卡那霉素对愈伤组织分化阶段的敏感程度比诱导愈伤阶段高;分化阶段选择压力要控制在25 mg/L以下;延迟加入卡那霉素或在分化阶段降低卡那霉素浓度是对筛选转化苗的可取办法。[结论]该研究为利用农杆菌介导法转化紫花苜蓿奠定了基础。

[Objective] The aim of the research was to study the sensitivity of Medicago sativa L leaves to kanamycin in the period of callus induction and differentiation.[Method] Effect of kanamycin on callus formation of M.sativa leaves was confirmed and its effect on callus differentiation was also ascertained based on callus formation.[Result] Different concentrations of kanamycin had some restraining effects on callus inducement and differentiation in M.sativa leaves,and the restraining effect enhanced along with the rising concentration of kanamycin.The sensitivity of kanamycin to callus in differentiation phase was higher than that in induction phase.The selective pressure should be controlled under 25 mg/L in differentiation phase.Delaying the adding of kanamycin or reducing the concentration of kanamycin in differentiation phase were advisable methods for screening transformation seedlings.[Conclusion] This study established the foundation for using agrobacterium-mediated method in transforming M.sativa.