The effect of feeding culture media with biogenetic precursors on high production of depsides in agitated shoot cultures of black and red aronias

Plant Cell, Tissue and Organ Culture (PCTOC) https://doi.org/10.1007/s11240-020-01869-4 ORIGINAL ARTICLE The effect of feeding culture media with biogenetic precursors on high production of depsides in agitated shoot cultures of black and red aronias Agnieszka Szopa1 · Paweł Kubica1 · Łukasz Komsta2 · Aleksandra Walkowicz‑Bożek1 · Halina Ekiert1 Received: 16 January 2020 / Accepted: 11 June 2020 © The Author(s) 2020 Abstract Agitated shoot cultures of two aronias, Aronia melanocarpa (Michx.) Elliott and Aronia arbutifolia (L.) Pers., were maintained on Murashige & Skoog medium (1 mg/l BA and 1 mg/l NAA), both with and without the addition of various biosynthetic precursors of phenolic acids and depsides (phenylalanine, cinnamic acid, benzoic acid and caffeic acid). Each substance was added in 5 concentrations (0.1–10 mmol/l), each concentration at two time points (at the beginning and on the 10th day of cultures). Twenty-four phenolic acids were determined in methanolic extracts of the biomasses collected after 20 days of growth cycles by means of HPLC method with DAD detection. The presence of seven compounds was confirmed in all the extracts—five depsides (neochlorogenic, chlorogenic, cryptochlorogenic, isochlorogenic and rosmarinic acids), and syringic and caffeic acids. The main metabolites in A. melanocarpa shoot extracts were isochlorogenic, chlorogenic and neochlorogenic acids (max. 249.88, 450.35, 192.16 mg/100 g DW). The main metabolites in A. arbutifolia shoot extracts were: chlorogenic, isochlorogenic and cryptochlorogenic acids (max. 361.60, 224.5, 526.2 mg/100 g DW). The largest total amounts of the compounds were confirmed in the cultures of both aronias after the addition of cinnamic acid (989.79 and 661.77 mg/100 g DW, respectively) and caffeic acid (854.99 and 1098.46 mg/100 g DW, respectively) at concentrations of 5 mmol/l on 10th day of growth cycles. These maximum amounts were 3.41, 3.42, 2.95 and 5.67 times higher, respectively, than in the control cultures. This is the first report documenting the high production of depsides in shoot cultures of black and red aronias after feeding with their biosynthetic precursors. Key message The influence of feeding culture media with biogenetic precursors on high production of depsides in agitated shoot cultures of Aronia melanocarpa and Aronia arbutifolia was proved. Communicated by Wagner Campos Otoni. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01869-4) contains supplementary material, which is available to authorized users. * Agnieszka Szopa * Halina Ekiert 1 Department of Pharmaceutical Botany, Collegium Medicum, Jagiellonian University, ul. Medyczna 9, 30‑688 Kraków, Poland 2 Department of Medicinal Chemistry, Faculty of Pharmacy with Division of Medical Analytics, Medical University of Lublin, ul. Chodźki 4a, 20‑093 Lublin, Poland 13 Vol.:(0123456789) Plant Cell, Tissue and Organ Culture (PCTOC) Keywords In vitro cultures · Phenylalanine · Cinnamic acid · Benzoic acid · Caffeic acid · Phenolic acids Abbreviations BA 6-Benzyladenine NAA 1-Naphthaleneacetic acid MS Murashige and Skoog HPLC-DAD High performance liquid chromatography with diode-array detector Introduction The pharmaceutical, cosmetics and health-food industry always look for new rich natural plant sources of antioxidants (Krishnaiah et al. 2011). This group of compounds possesses very valuable biological properties—they prevent a lot of lifestyle diseases (such as strokes, heart attacks, and neurodegenerative diseases), and can also be used as antiaging agents (Young and Woodside 2001; Matkowski 2008; Krishnaiah et al. 2011). Aronia genus, a representative of the Rosaceae family, is a rich source of natural antioxidants (Kulling and Rawel 2008; Kokotkiewicz et al. 2010). Aronia melanocarpa (Michx.) Elliott (black aronia) and Aronia arbutifolia (L.) Pers. (red aronia) are plant species of North American origin, cultivated in Central European countries. The fruits of black aronia are more famous than a plant raw material. They are rich in various groups of antioxidants, almost all polyphenols, e.g. phenolic acids and anthocyanins, procyanidin B 1, and also in flavonoids of high antioxidant activity. The presence of caffeic acid and two depsides (chlorogenic and neochlorogenic acid) was confirmed in fruit extracts. Anthocyanins are represented by cyanidin derivatives (cyanidin-3-galactoside and cyanidin3-arabinoside), whereas flavonoids by quercetin and its glycosides—quercitrin and rutoside. The fruits are additionally rich in vitamins—C, E, K, folic acid, as well as B-group vitamins and organic acids (Zhao et al. 2004; Kokotkiewicz et al. 2010; Taheri et al. 2013; Szopa et al. 2017). The knowledge about the chemical composition of red aronia fruits is very poor (Taheri et al. 2013). The earlier phytochemical studies performed by our team documented high amounts of various subgroups of polyphenols, chlorogenic acid, neochlorogenic acid, cyanidin glycosides, quercetin, quercitrin, and rutoside. Our studies provided additionally evidence that the leaves of both aronias are also a rich source of various polyphenols (Szopa et al. 2017). Biotechnological studies with different types of in vitro cultures of both black and red aronias (callus and shoot cultures, and solid and agitated cultures) performed by our team documented very high biosynthetic potential of cells of the tested plants in in vitro culture conditions. They produced 13 especially high amounts of phenolic acids. We looked for the most beneficial conditions for the production of this group of antioxidants (Szopa et al. 2013, 2018b, c, d; Szopa and Ekiert 2014). Phenolic acids are not only strong antioxidants. They possess a lot of other valuable biological activities, such as cytotoxic, anti-inflammatory, immunostimulatory, hypolipaemic, hepatoprotective and antiaggregatory, and these properties are the main reason of our studies in this area (Cai et al. 2004; Itoh et al. 2010; Rosa et al. 2016). So far, we tested the production of phenolic acids in in vitro cultures of both aronias under various basal media composition, concentration of plant growth regulators and light conditions (monochromatic and multispectral lights), as well as the type of culture (solid and agitated cultures) (Szopa et al. 2013; Szopa and Ekiert 2014; Szopa et al. 2018c, b). In the current part of investigation, we decided to investigate the effects of feeding culture media with biosynthetic precursors of phenolic acids. Our earlier results allowed us to choose one optimal variant of Murashige–Skoog (MS) medium (Murashige and Skoog 1962) enriched with 1 mg/l BA (6-benzyladenie) and 1 mg/l NAA (1-naphthaleneacetic acid) as a good universal “productive medium” for in vitro cultures of both aronias (Szopa et al. 2013, 2018d; Szopa and Ekiert 2014). This paper presents the effect of addition of biosynthetic precursors of phenol (...truncated)