外源物质对新疆紫草毛状根次生代谢产物含量的影响

Effects of Exogenous Substances on the Content of Secondary Metabolites in Arnebia euchroma Johnst Hairy Roots

目的:探讨不同外源物质对新疆紫草毛状根中次生代谢产物含量的影响。方法:采用分光光度法测定Ag NO3、L-苯丙氨酸、乙酸钠影响下新疆紫草毛状根提取物中次生代谢产物的含量。结果:Ag NO3质量浓度为5 mg/L时,毛状根中紫草素、总多酚及总黄酮的含量分别较对照提高了5、0.3倍和0.8倍,而花青素的含量在Ag NO3质量浓度为3 mg/L时达峰值,较对照提高了0.4倍。在Ag NO3质量浓度为5 mg/L前提下,L-苯丙氨酸质量浓度为25 mg/L时,紫草素、花青素及总黄酮的含量均达峰值,分别较对照提高了0.3、1.4倍和1.2倍。而总多酚含量虽然并未高于对照组,但是在各个处理中仍然含量最高。在Ag NO3、L-苯丙氨酸质量浓度分别为5、25 mg/L时,总多酚和紫草素含量均在乙酸钠质量浓度为50 mg/L时达到最高,花青素的含量在乙酸钠质量浓度为200 mg/L时达到最高,而乙酸钠不同处理的毛状根中总黄酮含量均明显低于对照。结论:外施一定质量浓度的诱导子Ag NO3及Ag NO3和L-苯丙氨酸的联合作用均能促进新疆紫草毛状根次生代谢产物的积累;外施一定质量浓度的Ag NO3、L-苯丙氨酸及乙酸钠可协同促进毛状根中总多酚、花青素和紫草素的积累,但显著抑制总黄酮的合成积累。

Objective： To investigate the effects of different exogenous substances on the content of secondary metabolites in Arnebia euchroma Johnst hairy roots and consequently provide a theoretical basis for further development and utilization of Arnebia euchroma Johnst hairy roots. Methods： The contents of secondary metabolites were measured by spectrophotometry from Arnebia euchroma Johnst hairy roots cultured in the presence of Ag NO3 alone and in combination with L-phenylalanine or L-phenylalanine plus sodium acetate. Results： The contents of shikonin, total polyphenols and total flavonoids in hairy roots cultured for 30 days at an Ag NO3 concentration of 5 mg/L respectively increased by 6, 1.3, 1.8 times compared with the control（CK）. In addition, when Ag NO3 concentration was 3 mg/L, the anthocyanin content reached its peak, a 1.4-fold increase over the control. Peak contents of shikonin, anthocyanins and total flavonoids were observed after culture in the presence of both 25 mg/L L-phenylalanine and 5 mg/L Ag NO3, which were increased by 1.3, 2.4 and 2.2 times, respectively, compared with the CK. The content of total polyphenols was not higher than that in the CK, but it was still the highest among three treatment groups. The contents of shikonin and total polyphenols reached their highest levels at 50 mg/L among different sodium acetate concentrations combined with 5 mg/L Ag NO3 and 25 mg/L L-phenylalanine, whereas the highest content of anthocyanin was observed when the sodium acetate concentration was 200 mg/L. The content of total flavonoids in hairy roots with sodium acetate treatments at various concentrations was significantly lower than that of the control. Conclusion： The addition of Ag NO3 and L-phenylalanine can cooperatively promote the accumulation of secondary metabolites in Arnebia euchroma Johnst hairy roots. Meanwhile, Ag NO3, L-phenylalanine and sodium acetate can cooperatively promote the accumulation of shikonin, polyphenol and anthocyanin, but significantly inhibit the accumulation of total flavonoids.