[关键词]
[摘要]
目的建立莨菪Hyoscyamus niger的发根培养体系。方法分别用A4、LBA9402和C58C1三种发根农杆菌菌株转化莨菪子叶受体,获得发根;测定发根的生长曲线和比较不同因素对发根生长量的影响;PCR鉴定转化发根;莨菪发根中生物碱类含量的测定。结果首次利用发根农杆菌A4、LBA9402、C58C1三种菌株成功从莨菪中诱导出发根,并建立了莨菪发根最优的培养体系;经PCR鉴定证明Ri质粒的T-DNA已转化进莨菪的发根中。结论莨菪发根培养体系的建立为利用遗传代谢工程手段调节莨菪次生代谢物含量和进行药用活性成分的工业化生产奠定了基础。
[Key word]
[Abstract]
Objective To establish the cultural system for hairy root of Hyoscyamus niger. Methods Hairy root of H. niger was obtained from cotyledon explants after infected with Agrobacterium rhizogenes strains A4, LBA9402, and C58C1, respectively. The growth curves were determined and the effects of different factors on the growth of hairy root were compared. The transformation of Ri T-DNA was examined by PCR. And the contents of alkaloids in hairy root were determined. Results The hairy root of H. niger was successfully induced by using A. rhizogenes strains A4, LBA9402, and C58C1 for the first time and the best system of hairy root culture was established. The transformation of T-DNA from Ri plasmid to the hairy root was confirmed by PCR analysis. Conclusion The establishment of cultural system for hairy root of H. niger lays foundation for regulating the secondary metabolism of H. niger by genetic engineering technique and for the large-scale production of active drug components.
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[基金项目]
江苏省药用植物生物技术重点实验室开放基金(KJS03080)