Production of Secondary Metabolites from Cell Cultures of Sageretia thea (Osbeck) M.C. Johnst. Using Balloon-Type Bubble Bioreactors

Abstract

Sageretia thea is used in the preparation of herbal medicine in China and Korea; this plant is rich in various bioactive compounds, including phenolics and flavonoids. The objective of the current study was to enhance the production of phenolic compounds in plant cell suspension cultures of Sageretia thea. Optimum callus was induced from cotyledon explants on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D; 0.5 mg L−1), naphthalene acetic acid (NAA, 0.5 mg L−1), kinetin (KN; 0.1 mg L−1) and sucrose (30 g L−1). Browning of callus was successfully avoided by using 200 mg L−1 ascorbic acid in the callus cultures. The elicitor effect of methyl jasmonate (MeJA), salicylic acid (SA), and sodium nitroprusside (SNP) was studied in cell suspension cultures, and the addition of 200 µM MeJA was found suitable for elicitation of phenolic accumulation in the cultured cells. Phenolic and flavonoid content and antioxidant activity were determined using 2,2 Diphenyl 1 picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethybenzothiazoline-6-sulphonic acid (ABTS), ferric reducing antioxidant power (FRAP) assays and results showed that cell cultures possessed highest phenolic and flavonoid content as well as highest DPPH, ABTS, and FRAP activities. Cell suspension cultures were established using 5 L capacity balloon-type bubble bioreactors using 2 L of MS medium 30 g L−1 sucrose and 0.5 mg L−1 2,4-D, 0.5 mg L−1 NAA, and 0.1 mg L−1 KN. The optimum yield of 230.81 g of fresh biomass and 16.48 g of dry biomass was evident after four weeks of cultures. High-pressure liquid chromatography (HPLC) analysis showed the cell biomass produced in bioreactors possessed higher concentrations of catechin hydrate, chlorogenic acid, naringenin, and other phenolic compounds.

Production of phenolic compounds, flavonoids, and lupeol by Lycium schweinfurthii suspension cultures in shake flasks and a rocking-motion bioreactor

“…Consequently, the PGRs concentration as well as the mineral content ensure the reliability, e ciency, and regeneration of the cell suspension initiation protocol(Rani et al., 2022; Schmitz et al, 2016). Based on the results obtained in our experiment and supported by the results obtained by other researchers (Binte Mosta z et al, 2018; Das et al, 2018;Kim et al, 2023), it is possible to summarize that the optimal concentration of NAA and 2,4-D for biomass growth of L. schweinfurthii is 1 mg L − 1 for each.In agreement with the current study,Nomura et al (2021) considered the SCV of a maximum of 20% as a starting cell density in Bamboo suspension cultures. Furthermore, the SCV of embryogenic cell suspension cultures of Pinus pinea reached between 50-90% by the end of cultivation in 100 and 200 mL shake asks (González-Cabrero et al, 2018).…”

“…The careful selection of appropriate explants, medium, and growth regulators plays a critical role in the initiation of cell cultures (Das et al, 2018). Also, any in vitro culture protocol needs to be customized for each plant species (Das et al, 2018;Kim et al, 2023). Commonly used arti cial media for callus and cell cultures include Murashige and Skoog (MS) medium, Gamborg B5 medium, and Linsmaier and Skoog (LS) medium (Binte Mosta z et al, 2018; Phillips et al, 2019;Ramachandra Rao et al, 2002).…”

“…2,4-D for the induction and maintenance of callus and cell cultures has been proven in many plant species such as rice (Binte Mosta z et al, 2018), Brucea mollis(Das et al, 2018) and Sageretia thea(Kim et al, 2023). The highest biomass of rice cells was obtained in a medium containing 3 mg L − 1 2,4-D with more than 80% cells induction frequency while NAA had a higher effect (100% induction frequency) compared to 2,4-D (93% induction frequency) for the production of Brucea mollis cells (Binte Mosta z et al, 2018;Das et al, 2018).…”

“…The highest biomass of rice cells was obtained in a medium containing 3 mg L − 1 2,4-D with more than 80% cells induction frequency while NAA had a higher effect (100% induction frequency) compared to 2,4-D (93% induction frequency) for the production of Brucea mollis cells (Binte Mosta z et al, 2018;Das et al, 2018). Otherwise, an equal concentration of NAA and 2,4-D (0.5 mg L − 1 each) was optimal for the initiation and production of Sageretia thea cell cultures(Kim et al, 2023). Moreover, the optimization of the PGRs concentration of 2,4-D, NAA, and kinetin to 1.33, 1.76, and 0.15 mg L − 1 , respectively, resulted in higher cell biomass of 70.62 g L − 1 DW of soybean cell cultures(Rani et al, 2022).…”

Metabolomics Reveals Rubiadin Accumulation and the Effects of Methyl Jasmonate Elicitation in Damnacanthus major Calli

“…Through in vitro callus cultures of D. major treated with MeJA, we successfully produced rubiadin, which is a valuable metabolite. In addition to the present findings, MeJA elicitation has also been employed to generate phenolic compounds in various plant cell cultures such as Thevetia peruviana [ 21 ], Sageretia thea [ 27 ], and Cnidium officinale [ 28 ]. Phenolic compounds have attracted considerable attention in culture systems in vitro because they can manipulate biosynthetic routes and accumulate the desired metabolites [ 29 ].…”