生物与非生物诱导子协同作用对丹参毛状根培养生产丹参酮的影响

Synergistic effects of biotic and abiotic elicitors on the production of tanshinones in Salvia miltiorrhiza hairy root culture

目的:考察生物诱导子yeast extract和不同非生物诱导子(Ag+,Co2+,α-氨基异丁酸)协同作用对丹参毛状根培养生产丹参酮的影响。方法:在丹参毛状根培养基中添加不同诱导子及诱导子组合,利用高效液相色谱法检测3种主要丹参酮(隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA)的含量。结果:yeast extract与各非生物诱导子在不同浓度下的组合对丹参酮Ⅰ和丹参酮ⅡA都发挥了较好的协同诱导作用,而对隐丹参酮的协同诱导效果不显著。其中,yeast extract和Ag+(300μmol.L-1)协同作用获得了最高的丹参酮Ⅰ产量,是对照组含量的13.9倍,诱导协同作用系数为3.0。yeast extract和Co2+(100μmol.L-1)协同作用获得了最高的丹参酮ⅡA产量,是对照组含量的14.5倍,协同作用系数为2.1。yeast extract和α-氨基异丁酸(200μmol.L-1)的组合是唯一一个比单个诱导子更有效地提高了隐丹参酮产量的组合,隐丹参酮的产量达到1.28 mg.g-1,是对照组含量的30.3倍,诱导协同作用系数为1.3。结论:利用生物诱导子yeast extract和不同的非生物诱导子进行协同作用,其诱导效果比单独使用某一种诱导子的诱导效果更好,这种相互作用可以更有效地促进次生代谢产物的合成。

Objective： To investigate the synergistic effects of a biotic elicitor yeast extract and different abiotic elicitors （Ag^＋ , Co^2＋ and α-amino isobutyric acid） on the production of tanshinones in Salvia miltiorrhiza hairy root. Method： Different elicitors and their combinations were added to S. miltiorrhiza hairy root culture and the contents of three major tanshinones （crypotanshinone, tanshinone I and tanshi- none ⅡA） were analyzed by HPLC. Result： The combinations of yeast extract with different abiotic elicitors had notable synergistic effects on the tanshinone Ⅰ and tanshinone ⅡA preduction but not on crypotanshinone. The combination of yeast extract and Ag^＋ （300 μmol·L^-1 ） yielded the highest tanshinone I content, which was nearly 14-fold of the control, and the synergistic elicitation coefficient was 3.0. The combination of yeast extract and Co^2＋ （100μmol·L^-1 ） led to the highest tanshinone IIs content, which was about 14.5-fold of the control, and the synergistic elicitation coefficient was 2.1. Only yeast extract combined with α-amino isobutyric acid （200 μmol·L^-1 ） increased the crypotanshinone content more effectively than single elicitors. The highest crypotanshinone content was 1.28mg·g^-1, about 30-fold of the control with a synergistic elicitation coefficient of 1.3. Conclusion： The elicitation by the combination of a biotic elicitor and an abiotic elicitor can generate a synergistic effect, which is more effective than single elicitors to promote secondary metabolite production in plant tissue cultures.