Abstract
Hairy root cultures are genetically and biochemically stable, and they regularly possess the same or better biosynthetic capabilities for specialized (secondary) metabolite production compared to the intact plant. Ononis species are well-known herbal remedies in ethnopharmacology and rich sources of isoflavonoids. Besides isoflavones, less prevalent isoflavones and pterocarpans with valuable biological effects can be found in Ononis species as well. As these plants are only collected but not cultivated, biotechnological methods could play a role in the larger-scale extraction of Ononis isoflavonoids. Regarding this information, we aimed to establish Ononis spinosa and Ononis arvensis hairy root cultures (HRCs) and analyze the isoflavonoid profile of hairy root cultures qualitatively and quantitatively, in order to define their capacity to produce biologically valuable isoflavonoids. During the qualitative description, beside isoflavonoids, two new phenolic lactones, namely, bulatlactone 2″-O-β-D-glucoside and ononilactone, were isolated, and their structures were characterized for the first time. Altogether, 29 compounds were identified by the means of UPLC-Orbitrap-MS/MS. Based on UHPLC-UV-DAD measurements, the isoflavonoid spectrum of the Ononis HRCs differed markedly from wild-grown samples, as they produce a limited range of the scaffolds. The most abundant compounds in the HRCs were medicarpin glucoside and sativanone glucoside. The overall isoflavonoid production of the cultures was comparable to wild-grown O. arvensis and approximately twice as high as in wild-grown O. spinosa samples. As the overall content of wild-grown samples include more isoflavonoid derivatives, the HRCs contain structurally less divergent isoflavonoids but in higher quantity.
Highlights
Bioactive natural products are molecules perfected by evolution, which, based on their physicochemical properties, are much more likely to become potential drug candidates than synthetic compounds produced by combinatorial chemical methods (Larsson et al, 2005)
High-performance liquid chromatography (HPLC) and Mass spectrometry (MS)-grade methanol and acetonitrile were purchased from Fischer Scientific Co. (Fair Lawn, NJ, United States); LiChropur formic- and acetic acid were obtained from Merck (Darmstadt, Germany)
In the first 2 weeks, the increase was lighter in the case of O. arvensis
Summary
Bioactive natural products are molecules perfected by evolution, which, based on their physicochemical properties, are much more likely to become potential drug candidates than synthetic compounds produced by combinatorial chemical methods (Larsson et al, 2005) Their sources might be herbs that are easy to cultivate, e.g., lavender or chamomile, etc., but in many cases, it Phytochemical Analysis of Ononis HRCs is only possible to isolate it from wild-growing populations, e.g., taxol. The reasons can lie in the difficulty of industrial upscaling Another problem is that the lack of differentiation and compartmentalization, that can result in different metabolic profiles compared to the intact plant (Atanasov et al, 2015). As opposed to cell cultures, they are genetically and biochemically stable, and they regularly possess the same or better biosynthetic capabilities for specialized metabolite production as the intact plant (Georgiev et al, 2012)
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