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目的研究紫锥菊毛状根的诱导及活性成分的产生。方法用发根农杆菌(Agrobacterium rhizogenes)A4,R1601,1025感染紫锥菊外植体。结果紫锥菊外植体被3种发根农杆菌感染后,外植体伤口处均能陆续分化生长出白色的毛状根。A4菌株的诱导效果明显好于其他2种菌株,A4对紫锥菊叶片的毛状根诱导率高达66.7%。紫锥菊毛状根的最佳诱导条件为:发根农杆菌A4,感染时间12 min,菌液A600值0.8,共培养温度24℃,共培养时间2 d,预培养时间2 d,培养基pH值6.0,此条件下紫锥菊毛状根的平均诱导率达到54%;对诱导出的毛状根进行PCR检测,证明其确为已转化的毛状根。检测得到紫锥菊毛状根中的多糖含量为15.54%,总酚的含量为2.491%。结论本实验所建立的紫锥菊毛状根培养系统,为研究紫锥菊毛状根大量培养生产活性成分奠定了基础。
Aim To study the induction of hairy roots and the generation of active components in Echinacea purpurea. Methods Agrobacterium rhizogenes A4, R1601 and 1025 were used to infect Echinacea explants. Results Echinacea explants were infected with three kinds of Agrobacterium rhizogenes, the hairy roots of the explants were differentiated and grew to white hairy roots. The induction effect of A4 strain was obviously better than the other two strains. A4 induced the hairy root induction rate of Echinacea purpurea as high as 66.7%. Echinacea hairy roots of the best induction conditions: Agrobacterium rhizogenes A4, infection time 12 min, the A600 value 0.8, co-culture temperature 24 ℃, co-culture time 2 d, pre-culture time 2 d, medium pH 6.0. Under these conditions, the average induction rate of Echinacea hairy roots reached 54%. The induced hairy roots were detected by PCR and proved to be transformed hairy roots. The content of polysaccharide in Echinacea hairy roots was 15.54% and the total phenolic content was 2.491%. Conclusion The hairy root culture system of Echinacea purpurea established in this experiment lays the foundation for the research on the active cultivation of Echinacea purpurea hairy roots in large quantities.