红花悬浮细胞体系的建立及其化学成分的UPLC-Q-TOF/MS分析

Establishment of cell suspension culture system of Carthamus tinctorius and analysis of compounds by UPLC-Q-TOF/MS

目的构建红花悬浮细胞体系,为进一步研究红花的功能基因搭建平台。方法用红花无菌子叶作为外植体进行愈伤组织培养,选择淡黄色、质地疏松、增殖速度快的愈伤组织为材料进行悬浮培养,考察激素浓度、愈伤起始接种量、p H值、细胞活力对红花悬浮细胞体系的影响以及不同浓度茉莉酸甲酯（Me JA）刺激对悬浮细胞生长曲线的变化。采用UPLC-Q-TOF/MS法对红花悬浮细胞培养体系进行初步化学成分分析。结果红花悬浮细胞培养的最适宜配方是MS＋1.0 mg/L TDZ＋0.1 mg/L NAA,最适宜p H值为5.5~6.0,最适宜接种量为0.02 g/m L培养体系;不同浓度的Me JA刺激对细胞增殖率影响较大,50μmol/L Me JA能提高悬浮细胞增殖率,500μmol/L Me JA能对细胞增殖则有明显的抑制作用。从红花悬浮细胞培养体系中初步鉴定出13种化合物。结论利用红花子叶能建立较好的悬浮细胞培养体系,适宜浓度的Me JA刺激可提高悬浮细胞增殖率。

Objective To study the establishment of cell suspension culture system and build a platform for exploring the functional genes in the biosynthesis pathway for Carthamus tinctorius. Methods Using the cotyledon of C. tinctorius as explants for callus induction, callus with good quality was selected for suspension cultivation. The factors influencing the establishment of C. tinctorius cell suspension culture system were investigated, such as hormone, inoculum amount, p H, and cell activity. The effect of Me JA at different concentration on suspension cell growth rate was studied. Chemical compounds in suspension cell system were preliminary identified by UPLC-Q-TOF/MS. Results The optimal medium for the cell suspension culture was MS ＋ 1.0 mg/LTDZ ＋ 0.1 mg/L NAA, the p H was 5.5—6.0, and inoculum amount was 0.02 g/m L. Me JA at different concentration had effects on suspension cell growth rate： 50 μmol/L Me JA increased cell growth rate, 500 μmol/L Me JA depressed cell growth rate greatly. Thirteen potential chemical compounds in suspension cell system were preliminary identified. Conclusion The cell suspension culture system of C. tinctorius is established, and Me JA with optimal concentration has effects on suspension cell growth rate.