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本研究从设计多种诱导条件出发,探索建立稳定的柴胡毛状根诱导体系。发根农杆菌A4侵染北柴胡无菌苗叶基部,共培养3天后,转入抑菌培养,10天开始出现毛状根,经4～5周培养,转为液体摇培,获得大量毛状根,诱导产生的毛状根可再生成植株。毛状根再生成植株有两种方式:一种是液体摇培的毛状根自发脱落形成幼苗的继续培养,另一种是将毛状根产生脱落的类愈伤组织置于经筛选的再生固体培养基上再生成植株。柴胡毛状根及其植株再生体系的建立,为开展柴胡次生代谢研究,尤其是利用农杆菌介导的遗传转化研究和利用奠定了基础。由毛状根诱导出再生植株在新种质产生和品种改良选育方面具有应用价值。
In this study, starting from the design of a variety of conditions to explore and establish a stable Bupleurum hairy root induction system. Agrobacterium rhizogenes A4 invaded the base of Bupleurum chinense L. aseptic seedling leaves and co-cultured for 3 days. After being cultured in bacteriostatic culture for 10 days, hairy roots began to appear after 10 days. After 4 to 5 weeks of culture, Hairy roots, induced hairy roots can be regenerated into plants. Hairy roots regenerate into plants in two ways: one is the spontaneous shedding of the hairy roots of the liquid to form the seedlings, and the other is that the callus, which is shed by the hairy roots, is placed in the regenerated Regenerated plants on solid medium. The establishment of Bupleurum chinense hairy roots and their plant regeneration system lays a foundation for the research on the secondary metabolites of Bupleurum chinense, especially using Agrobacterium-mediated transformation and utilization. Induced by hairy roots of regenerated plants in the new germplasm generation and breeding of improved varieties has application value.