Cai G, Li G, Ye H, Li G
Biology Department of Beijing Normal University, China.
Chin J Biotechnol. 1995;11(4):227-35.
The hairy root culture system of the medical plant Artemisia annua L. was established by infection with Agrobacterium rhizogenes R1601. The transgenic state of transformed roots was confirmed by Southern blot hybridization with TL-DNA of pFw302. The expression of NPTII gene was confirmed by enzymic assay. The important secondary metabolites-artemisinin was obtained in the hairy root culture. The effects of various physical and chemical factors on the growth of the hairy roots and production of artemisinin were studied. Artemisinin could be detected in hairy roots cultures in the light. The optimum pH value of the medium was 5.4. Fast growth of the hairy roots and maximal production of artemisinin was observed in the presence of 3% sucrose. Low concentration of naphthylacetic acid (0.025 mg/L) enhanced the growth of the roots but inhibited the production of artemisinin. The growth and artemisinin production in hairy root cultures were greatly promoted by the addition of gibberellin (GA3) to the medium. Its optimum concentration was 4.8 mg/L.
通过发根农杆菌R1601感染建立了药用植物青蒿的毛状根培养体系。用pFw302的TL-DNA进行Southern杂交，证实了转化根的转基因状态。通过酶法测定证实了NPTII基因的表达。在毛状根培养中获得了重要的次生代谢产物青蒿素。研究了各种物理和化学因素对毛状根生长及青蒿素产量的影响。在光照条件下，毛状根培养物中可检测到青蒿素。培养基的最适pH值为5.4。在3%蔗糖存在下，观察到毛状根快速生长且青蒿素产量最高。低浓度的萘乙酸（0.025 mg/L）促进根的生长，但抑制青蒿素的产生。向培养基中添加赤霉素（GA3）极大地促进了毛状根培养物的生长和青蒿素的产生。其最适浓度为4.8 mg/L。
