真菌诱导子与吸附树脂对新疆紫草毛状根中萘醌积累的影响

Influence of fungal elicitor and macroporous resin on shikonin accumulation in hairy roots of Arnebia euchroma (Royle) Johnst

摘要:

通过考察真菌诱导子与吸附树脂对新疆紫草毛状根中萘醌积累的影响，获得真菌诱导子与吸附树脂对萘醌类物质积累的最佳处理，为规模化生产提供依据。以新疆紫草毛状根为试验材料，将黑曲霉、米曲霉诱导子及其混合诱导子、大孔吸附树脂添加到M-9培养基中，采用分光光度法测定毛状根总萘醌含量。试验结果表明：在毛状根培养10 d时以2.5∶50的比例添加混合诱导子，总萘醌含量是对照的2.28倍；在此结果基础上，在培养第0天添加大孔吸附树脂NKA-9，总萘醌含量最高是对照的3.71倍；黑曲霉诱导子与米曲霉诱导子有协同效应；在生物反应器中添加混合诱导子及大孔吸附树脂NKA-9，其总萘醌含量是对照的4.17倍。米曲霉诱导子、混合诱导子对毛状根增殖有促进作用；同时添加大孔吸附树脂NKA-9及混合诱导子能提高毛状根总萘醌含量。生物反应器培养毛状根为今后利用新疆紫草毛状根规模化生产总萘醌提供了理论参考。

Abstract:

In order to investigate the effects of fungal elicitor and macroporous adsorption resin on shikonin accumulation in hairy roots of arnebia euchroma (Royle) Johnst, we used spectrophotometry to determine the total naphthoquinone content of the hairy roots, by adding different volume ratio of Aspergillus niger elicitor, Aspergillus oryzae elicitor, and the macroporous resin into the M-9 liquid medium at different culture time. The results show that the total naphthoquinone content was 2.28 times higher than the control when we added mixed elicitors of Aspergillus niger and Aspergillus oryzae at the ratio of 2.5:50 in the 10th day of hairy roots cultivating. The total naphthoquinone content was 3.71 times higher than that of the control, when we added macroporous adsorption resin NKA-9. Aspergillus niger elicitor exhibited synergistic effect with Aspergillus oryzae elicitor to enhance the naphthoquinone. Also, the total naphthoquinone level was 4.17 times higher than that of the control by adding mixed fungal elicitor and macroporous adsorption resin NKA-9 in the bioreactor. Aspergillus oryzae and mixed elicitor could promote the hairy roots proliferation, and macroporous adsorption resin NKA-9 and mixed elicitor increased the total naphthoquinone content. In summary, the measure developed for Arnebia euchroma (Royle) Johnst hairy roots cultivating in bioreactors may potential for large-scale production of naphthoquinone.