Titova Maria V, Lunkova Maria K, Tyurina Tatiana M, Prudnikova Olga N, Popova Elena V, Klychnikov Oleg I, Metalnikov Pavel S, Ikhalaynen Yuri A, Vasileva Elizaveta N, Rodin Igor A, Nosov Alexander M
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia.
Department of Biochemistry, Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia.
Front Plant Sci. 2024 Feb 26;15:1349494. doi: 10.3389/fpls.2024.1349494. eCollection 2024.
is a valuable medicinal plant and a source of a broad spectrum of biologically active ginsenosides of different structural groups. Overexploitation and low adaptability to planation cultivation have made this species vulnerable to human pressure and prompted the development of cell cultivation as a sustainable alternative to harvesting wild plants for their bioactive components. Despite high interest in biotechnological production, little is known about the main factors affecting cell growth and ginsenoside biosynthesis of this species under conditions. In this study, the potential of cell cultures of as a biotechnological source of ginsenosides was was assessed.
Six suspension cell lines that were developed from different sections of a single rhizome through a multi-step culture optimization process and maintained for over 3 years on media with different mineral salt base and varying contents of auxins and cytokinins. These cell lines were evaluated for productivity parameters and cytological characteristics. Ginsenoside profiles were assessed using a combination of the reversed-phase ultra-high-performance liquid chromatography-Orbitrap-tandem mass spectrometry (UHPLC-Orbitrap-MS/MS) and ultra-performance liquid chromatography-time of flight-mass spectrometry (UPLC-TOF-MS).
All lines demonstrated good growth with a specific growth rate of 0.1-0.2 day, economic coefficient of 0.31-0.70, productivity on dry weight (DW) of 0.30-0.83 gDW (L·day), and maximum biomass accumulation varying from 10 to 22 gDW L. Ginsenosides of the protopanaxadiol (Rb1, Rb2/Rb3, malonyl-Rb1, and malonyl-Rb2/Rb3), oleanolic acid (R0 and chikusetsusaponin IV), and ocotillol (vinaginsenoside R1) groups and their isomers were identified in cell biomass extracts. Chikusetsusaponin IV was identified in cell culture for the first time.
These results suggest that suspension cell cultures of Vietnamese ginseng have a high potential for the biotechnological production of biomass containing ginsenosides, particularly of the oleanolic acid and ocotillol groups.
越南人参是一种珍贵的药用植物，是不同结构基团的多种具有生物活性的人参皂苷的来源。过度开发以及对种植栽培的低适应性使得该物种易受人类活动压力影响，并促使细胞培养作为一种可持续的替代方法得以发展，以取代从野生植物中获取其生物活性成分。尽管对生物技术生产有很高的兴趣，但对于影响该物种细胞生长和人参皂苷生物合成的主要因素在特定条件下的了解却很少。在本研究中，评估了越南人参细胞培养物作为人参皂苷生物技术来源的潜力。
通过多步培养优化过程从单个根茎的不同部分建立了六个悬浮细胞系，并在含有不同矿质盐基础以及不同生长素和细胞分裂素含量的培养基上维持了3年以上。对这些细胞系的生产力参数和细胞学特征进行了评估。使用反相超高效液相色谱-轨道阱串联质谱（UHPLC-Orbitrap-MS/MS）和超高效液相色谱-飞行时间质谱（UPLC-TOF-MS）相结合的方法评估人参皂苷谱。
所有细胞系均表现出良好的生长，比生长速率为0.1 - 0.2天，经济系数为0.31 - 0.70，干重（DW）生产力为0.30 - 0.83 gDW/（L·天），最大生物量积累在10至22 gDW/L之间。在细胞生物量提取物中鉴定出了原人参二醇（Rb1、Rb2/Rb3、丙二酰-Rb1和丙二酰-Rb2/Rb3）、齐墩果酸（R0和竹节人参皂苷IV）、奥克梯隆醇（越南人参皂苷R1）基团及其异构体的人参皂苷。竹节人参皂苷IV首次在越南人参细胞培养物中被鉴定出来。
这些结果表明，越南人参悬浮细胞培养物在生物技术生产含人参皂苷的生物量方面具有很高的潜力，特别是齐墩果酸和奥克梯隆醇基团的人参皂苷。
