洛伐他丁、氯胺和茉莉酸甲酯处理对茜草悬浮培养细胞中紫色素和大叶茜草素积累的作用

摘要

目的:使用茜草细胞悬浮培养体系研究茜草中萘醌类成分生合成途径原料IPP的来源,以及茉莉酸甲酯、氯胺和洛伐他丁对蒽醌和萘醌的积累情况.方法:建立茜草细胞悬浮培养体系,分别在培养基中添加茉莉酸甲酯、氯胺和洛伐他丁.每3天取样一次.使用高效液相色谱法测定紫色素和大叶茜草素含量并计算出产量.结果:洛伐他丁处理组中紫色素和大叶茜草素产量均有显著增加.氯胺处理组中紫色素和大叶茜草素产量明显下降.茉莉酸甲酯处理组中紫色素产量增加,同时大叶茜草素产量降低.结论:在茜草细胞中萘醌类成分的IPP可能是通过MEP途径合成的；要解释茉莉酸甲酯对蒽醌和萘醌积累的不同作用,基因表达水平方面的研究尤其是对1,4-二羟基-2-萘甲酸异戊烯基化的基因表达研究应该受到更多关注.%AIM:To determine the IPP origin of the naphthoquinones (NQs) in Rubia cordifolia,and to evaluate the effects of methyl jasmonate (MeJA) treatment,MEP,and MVA pathway inhibitor treatment on the accumulation of anthraquinones (AQs) and NQs in cell suspension cultures of R.cordifolia.METHODS:Cell suspension cultures of R.cordifolia were established.Specific inhibitors (lovastatin and clomazone) and MeJA were supplied to the media,respectively.Treated cells were sampled every three days.Content determination of purpurin (AQs) and mollugin (NQs) were carried out using RP-HPLC.The yield of the two compounds was compared with the DMSO-supplied group and the possible mechanism was discussed.RESULTS:Lovastatin treatment increased the yield of purpurin and mollugin significantly.Clomazone treatment resulted in a remarkable decrease of both compounds.In the MeJA-treated cells,the purpurin yield increased,meanwhile,the mollugin yield decreased compared with control.CONCLUSION:the IPP origin of mollugin in R.cordifolia cell suspension cultures was likely from the MEP pathway.To explain the different effects of MeJA on AQs and NQs accumulation,studies on the regulation and expression of the genes,especially after prenylation of 1,4-dihydroxy-2-naphthoic acid should be conducted.