We combined machine learning and plant in vitro culture methodologies as a novel approach for unraveling the phytochemical potential of unexploited medicinal plants. In order to induce phenolic compound biosynthesis, the in vitro culture of three different species of Bryophyllum under nutritional stress was established. To optimize phenolic extraction, four solvents with different MeOH proportions were used, and total phenolic content (TPC), flavonoid content (FC) and radical-scavenging activity (RSA) were determined. All results were subjected to data modeling with the application of artificial neural networks to provide insight into the significant factors that influence such multifactorial processes. Our findings suggest that aerial parts accumulate a higher proportion of phenolic compounds and flavonoids in comparison to roots. TPC was increased under ammonium concentrations below 15 mM, and their extraction was maximum when using solvents with intermediate methanol proportions (55–85%). The same behavior was reported for RSA, and, conversely, FC was independent of culture media composition, and their extraction was enhanced using solvents with high methanol proportions (>85%). These findings confer a wide perspective about the relationship between abiotic stress and secondary metabolism and could serve as the starting point for the optimization of bioactive compound production at a biotechnological scale.

The increase in life expectancy has led to the appearance of chronic diseases and interest in healthy aging, in turn promoting a growing interest in bioactive compounds (BCs) and functional ingredients. There are certain foods or products rich in functional ingredients, and algae are one of them. Algae consumption has been nominal in Europe until now. However, in recent years, it has grown significantly, partly due to globalization and the adoption of new food trends. With the aim of obtaining BCs from foods, multiple methods have been proposed, ranging from conventional ones, such as maceration or Soxhlet extraction, to more innovative methods, e.g., ultrasound-assisted extraction (UAE). UAE constitutes a novel method, belonging to so-called green chemistry, that enables the extraction of BCs requiring lower amounts of solvent and energy costs, preserving the integrity of such molecules. In recent years, this method has been often used for the extraction of different BCs from a wide range of algae, especially polysaccharides, such as carrageenans and alginate; pigments, including fucoxanthin, chlorophylls, or β-carotene; and phenolic compounds, among others. In this way, the application of UAE to marine algae is an efficient and sustainable strategy to pursue their deep characterization as a new source of BCs, especially suitable for vegetarian and vegan diets.

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The rates of production of secondary metabolites obtained by employing conventional plant breeding may be low for practical purposes. Thus, innovative approaches for increasing their rates of production are being developed. Here, we propose the use of elicited plant suspension cultured cells (PSCC) with cyclodextrins (CDs) as an alternative method for the production of bioactive compounds from Bryophyllum species. For this purpose, we analyzed the effects of methyl–β–cyclodextrin and 2–hydroxypropyl–β–cyclodextrin on cell culture growth and on the intra- and extracellular production of phenols and flavonoids. Results clearly show that CDs enhance the biosynthesis of polyphenols by PSCC favoring their accumulation outside the cells. CDs shift the homeostatic equilibrium by complexing extracellular phenolics, causing stress in cells that respond by increasing the production of intracellular phenolics. We also analyzed the radical scavenging activity of the culture medium extracts against 2,2–diphenyl–1–pycrilhydrazyl (DPPH) radical, which increased with respect to the control samples (no added CDs). Our results suggest that both the increase in the production of polyphenols and their radical scavenging activity are a consequence of their inclusion in the CD cavities. Overall, based on our findings, CDs can be employed as hosts for increasing the production of polyphenols from Bryophyllum species.

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