Suspension cell cultures of Panax vietnamensis as a biotechnological source of ginsenosides: growth, cytology, and ginsenoside profile assessment

Maria V Titova; Maria K Lunkova; Tatiana M Tyurina; Olga N Prudnikova; Elena V Popova; Oleg I Klychnikov; Pavel S Metalnikov; Yuri A Ikhalaynen; Elizaveta N Vasileva; Igor A Rodin; Alexander M Nosov

doi:10.3389/fpls.2024.1349494

Suspension cell cultures of Panax vietnamensis as a biotechnological source of ginsenosides: growth, cytology, and ginsenoside profile assessment

Front Plant Sci. 2024 Feb 26:15:1349494. doi: 10.3389/fpls.2024.1349494. eCollection 2024.

Authors

Affiliations

¹ K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Moscow, Russia.
² Department of Biochemistry, Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia.
³ Department of Analytical Chemistry, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia.
⁴ Department of Plant Physiology, Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia.

^# Contributed equally.

Abstract

Introduction: Panax vietnamensis 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 in vitro 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 in vitro conditions. In this study, the potential of cell cultures of P. vietnamensis as a biotechnological source of ginsenosides was was assessed.

Methods: 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).

Results: All lines demonstrated good growth with a specific growth rate of 0.1-0.2 day^-1, economic coefficient of 0.31-0.70, productivity on dry weight (DW) of 0.30-0.83 gDW (L·day)^-1, and maximum biomass accumulation varying from 10 to 22 gDW L^-1. 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 P. vietnamensis cell culture for the first time.

Discussion: 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.

Keywords: Vietnamese ginseng; ginsenosides; plant biotechnology; plant cell culture; suspension cell culture.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Cultivation of cell culture in flasks and biochemical analysis were performed using the equipment of the large-scale research facility “All-Russian Collection of cell cultures of higher plants” (ARCCC HP IPPRAS) and “Experimental biotechnological facility” (EBF IPPRAS). Cell culture induction was financially supported within the state assignments of the Ministry of Science and Higher Education of the Russian Federation, theme No. 122042700045-3. Suspension cell line cultivation in flasks was financially supported within the state assignments of the Ministry of Science and Higher Education of the Russian Federation, theme No. 122042600086-7. Biochemical analysis of the extracts was performed under financial support of the Russian Science Foundation, project No. 19-14-00387.