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Plant cell cultures of various yew species are a profitable source of taxoids (taxane diterpenoids) with antitumor activity. So far, despite intensive studies, the principles of the formation of different groups of taxoids in cultured in vitro plant cells have not been fully revealed. In this study, the qualitative composition of taxoids of different structural groups was assessed in callus and suspension cell cultures of three yew species (Taxus baccata, T. canadensis, and T. wallichiana) and two T. × media hybrids. For the first time, 14-hydroxylated taxoids were isolated from the biomass of the suspension culture of T. baccata cells, and their structures were identified by high-resolution mass spectrometry and NMR spectroscopy as 7β-hydroxy-taxuyunnanin C, sinenxane C, taxuyunnanine C, 2α,5α,9α,10β,14β-pentaacetoxy-4(20), 11-taxadiene, and yunnanxane. UPLC–ESI-MS screening of taxoids was performed in more than 20 callus and suspension cell lines originating from different explants and grown in over 20 formulations of nutrient media. Regardless of the species, cell line origin, and conditions, most of the investigated cell cultures retained the ability to form taxane diterpenoids. Nonpolar 14-hydroxylated taxoids (in the form of polyesters) were predominant under in vitro culture conditions in all cell lines. These results, together with the literature data, suggest that dedifferentiated cell cultures of various yew species retain the ability to synthesize taxoids, but predominantly of the 14-OH taxoid group compared to the 13-OH taxoids found in plants.

Pollen–stigma interactions have been studied extensively because they play an important role in sexual reproduction and crop yield. The vast majority of studies have focused on dry stigmas, which are typical of many model and agricultural plants; however, the data obtained are difficult to apply to plants with wet stigmas, such as tomato and tobacco. Pollen germination in this case occurs in a liquid, an exudate, which has a complex, species-specific composition. UPLC-ESI-MS-based hormone screening was carried out for six plant genera belonging to Solanaceae, Bromeliaceae, and Gesneriaceae families and revealed jasmonic acid (JA), abscisic acid (ABA) and/or jasmonoyl-L-isoleucine (IleJA) in stigma exudates of tobacco, tomato, and Streptocarpus sp. To assess the physiological significance of plant hormones in stigma exudate we tested their effect in vitro, finding that JA, IleJA, and MeJa significantly stimulated germination of tobacco pollen, with JA being most effective in accordance with its predominance in the stigma exudate; furthermore, ABA stimulated pollen germination in all tested species including bromeliads despite the lack of this hormone in their exudates. Both JA and ABA had an anti-oxidant effect on germinating pollen. Possible functions of hormones and ROS in exudate as well as ways of implementing the anti-oxidant effect of phytohormones are discussed.

Dioscorea deltoidea is a medicinal plant valued for its high content of steroidal glycosides (SG)—bioactive compounds with cardioprotective and immunomodulation actions, also used to treat reproductive system disorders. To overcome the limitations of natural resources of this species, a suspension cell culture of D. deltoidea was developed as a renewable and ecologically sustainable source of raw biomass and SG. Cell culture demonstrated stable and intensive growth in the laboratory (20 L) and industrial (630 L) bioreactors operated under a semi-continuous regime (specific growth rate 0.11–1.12 day−1, growth index 3.5–3.7). Maximum dry weight accumulation (8.5–8.8 g/L) and SG content (47–57 mg/g DW) were recorded during the stationary phase. Bioreactor-produced cell biomass contained inorganic macro (K, Ca, Mg, Na) and micro (Zn, Mn, Fe, B, Al, Cu, Cr, Se, Co, Ni) elements in concentrations within the safe range of dietary recommendations. Acute toxicity test showed no or insignificant changes in organ weight, hematological panel and blood biochemistry of laboratory animals fed with 2000 and 5000 mg/kg dry biomass. The results suggest that cell culture of D. deltoidea grown in bioreactors has great potential to be used as functional foods and a component of specialized dietary supplements in complex therapy of reproductive system disorders and mineral deficiency.

Polyscias fruticosa (L.) Harms, or Ming aralia, is a medicinal plant of the Araliaceae family, which is highly valued for its antitoxic, anti-inflammatory, analgesic, antibacterial, anti-asthmatic, adaptogenic, and other properties. The plant can be potentially used to treat diabetes and its complications, ischemic brain damage, and Parkinson’s disease. Triterpene glycosides of the oleanane type, such as 3-O-[β-D-glucopyranosyl-(1→4)-β-D-glucuronopyranosyl] oleanolic acid 28-O-β-D-glucopyranosyl ester (PFS), ladyginoside A, and polysciosides A-H, are mainly responsible for biological activities of this species. In this study, cultivation of the cell suspension of P. fruticosa in 20 L bubble-type bioreactors was attempted as a sustainable method for cell biomass production of this valuable species and an alternative to overexploitation of wild plant resources. Cell suspension cultivated in bioreactors under a semi-continuous regime demonstrated satisfactory growth with a specific growth rate of 0.11 day−1, productivity of 0.32 g (L · day)−1, and an economic coefficient of 0.16 but slightly lower maximum biomass accumulation (~6.8 g L−1) compared to flask culture (~8.2 g L−1). Triterpene glycosides PFS (0.91 mg gDW−1) and ladyginoside A (0.77 mg gDW−1) were detected in bioreactor-produced cell biomass in higher concentrations compared to cells grown in flasks (0.50 and 0.22 mg gDW−1, respectively). In antibacterial tests, the minimum inhibitory concentrations (MICs) of cell biomass extracts against the most common pathogens Staphylococcus aureus, methicillin-resistant strain MRSA, Pseudomonas aeruginosa, and Escherichia coli varied within 250–2000 µg mL−1 which was higher compared to extracts of greenhouse plant leaves (MIC = 4000 µg mL−1). Cell biomass extracts also exhibited antioxidant activity, as confirmed by DPPH and TEAC assays. Our results suggest that bioreactor cultivation of P. fruticosa suspension cell culture may be a perspective method for the sustainable biomass production of this species.

Callus and suspension cell cultures of rare foxglove species, Digitalis ciliata and D. grandiflora, were induced from cotyledons and hypocotyls of in vitro seedlings, and their growth and phytochemical profiles were investigated. In both species, callus induction was more efficient from leaf explants (60–80%) than from hypocotyl explants (15–35%). Callus cultures of both species grew well with growth indices ranged from 5 to 10 depending on the genotype. Suspension culture growth profiles also differed between the two species with a 10–11-days lag-phase observed for D. grandiflora and a bi-phasic growth curve without lag-phase recorded for D. ciliata. The main growth characteristics of the D. grandiflora suspension cell culture (maximum biomass accumulation ~ 14 g/L, growth index ~ 10, economic coefficient ~ 0.42, biomass productivity ~ 0.53 g/(L × day)) were 1.5–3 times higher than those for D. ciliata. Ten compounds were identified in cell biomass using UPLC-ESI-Q-TOF-MS: phenylethanoids digiciliside A, digiciliside B, maxoside, purpureaside E, their methyl derivatives and isomers, and two furostanol glycosides with aglycone tigogenin. Phenylethanoid glycosides were major compounds and comprised 0.8–1.1% of dry weight. During the two-years cultivation, suspension cultures retained the ability to accumulate most of the identified compounds evidencing for stability of species-specific secondary metabolism in cultured foxglove cells during this period.Key messagePhenylethanoid glycosides (1.1% DW) and steroidal furostanol glycosides were identified in callus and suspension cell cultures of rare medicinal species Digitalis ciliata and D. grandiflora developed for the first time.

In the present study, we explored the therapeutic potential of bioreactor-grown cell cultures of the medicinal plant species Dioscorea deltoidea, Tribulus terrestris and Panax japonicus to treat carbohydrate metabolism disorders (CMDs) in laboratory rats. In the adrenaline model of hyperglycemia, aqueous suspensions of cell biomass pre-administered at a dose of 100 mg dry biomass/kg significantly reduced glucose level in animal blood 1–2.5 h (D. deltoidea and T. terrestris) or 1 h (P. japonicus) after adrenaline hydrochloride administration. In a streptozotocin-induced model of type 2 diabetes mellitus, the cell biomass of D. deltoidea and T. terrestris acted towards normalization of carbohydrate and lipid metabolism, as evidenced by a significant reduction of daily diuresis (by 39–57%), blood-glucose level (by 46–51%), blood content in urine (by 78–80%) and total cholesterol (25–36%) compared to animals without treatment. Bioactive secondary metabolites identified in the cell cultures and potentially responsible for their actions were deltoside, 25(S)-protodioscin and protodioscin in D. deltoidea; furostanol-type steroidal glycosides and quinic acid derivatives in T. terrestris; and ginsenosides and malonyl-ginsenosides in P. japonicus. These results evidenced for high potential of bioreactor-grown cell suspensions of these species for prevention and treatment of CMD, which requires further investigation.

Callus and suspension cell cultures were successfully developed from Sutherlandia frutescens (Fabaceae), an endemic medicinal plant of South Africa. Two callus cell lines, originating from hypocotyl and cotyledon explants of in vitro seedlings under both dark and light conditions, showed intensive fresh weight accumulation with growth index ranging from 4.6 to 5.9. Suspension cell cultures induced from two callus lines had similar growth profiles and their growth index (15–18), specific growth rate (0.15–0.16 day −1 ), productivity (0.83–0.96 g/(l day)) and maximum biomass accumulation (16–18 g/l) remained relatively high for Fabaceae cell cultures during 27 sub-cultivations. Callus and suspension cell cultures showed similar profiles of secondary metabolites that were, however, different from leaves of greenhouse plants. Isoflavones were predominant in both callus and suspension cell cultures while flavonoids (sutherlandins) and triterpene glycosides of the cycloartane group (sutherlandiosides) were mostly found in leaves. Nineteen fatty acids (FA), both short- and very-long-chained (up to C 25:0 ), were found in cell cultures. Linoleic and α-linolenic FA together comprised 60–64% out of total FA content in cell cultures followed by palmitic acid (18–25%). Extracts of suspension cell biomass exhibited antimicrobial activity against Staphylococcus aureus but were not effective against Pseudomonas aeruginosa. To the best of our knowledge, this is the first report on the induction, phytochemical composition and antimicrobial activity screening of S. frutescens suspension cell cultures which opens the door for their biotechnological application.