Micropropagation and HPLC-DAD, UPLC MS/MS analysis of oenothein B and phenolic acids in shoot cultures and in regenerated plants of fireweed (Chamerion angustifolium (L.) Holub)

Abstract

In this study, a micropropagation protocol using nodal explants from in vitro grown plants of Chamerion angustifolium (L.) Holub was developed and analysis of oenothein B and selected phenolic acids in shoot cultures was performed for the first time. For shoot induction and multiplication Murashige and Skoog’s (MS) basal medium supplemented with 2-isopentenyladenine (2iP), zeatin (Z) and 6-benzyloaminopurine (BAP) was used. 2iP was the most responsive in terms of promoting shoots per explant with the maximum (6.57 ± 1.14) recorded at a concentration of 2.0 mg L−1 after 6 weeks of culture. After two subcultures the multiplication rate was increased up to 19 shoots per explant on medium with 2iP (1.0 mg L−1). To prevent tissue browning, ascorbic acid and casein hydrolysate were added to the induction medium, resulting in a reduction of browning by 30%. The rooted plantlets were successfully transferred to soil and acclimatized with 97% frequency. Quantitative and qualitative assessments of oenothein B and phenolic acid contents in in vitro regenerated shoots as well as in ex vitro plants were performed using high-performance liquid chromatography with a diode-array detector (HPLC-DAD) and ultra-performance liquid chromatography–tandem mass spectrometry (UPLC MS/MS) methods. Oenothein B (1.62‒4.55 g 100 g−1 DW), ellagic acid, gallic and caffeic acids were identified in in vitro regenerated plants. The results of this study confirm that the oenothein B-producing plantlets can be obtained using the micropropagation method with axillary shoots being a valuable source of oenothein B and phenolic acids.

Phytochemical variability during vegetation of Chamerion angustifolium (L.) Holub genotypes derived from in vitro cultures

“…The micropropagation efficiency depends on culture conditions, selection of plant growth regulators (PGRs), micropropagation technique, type of explant, age of donors' plants and genotype. Organogenesis from root explants (Turker et al 2008;Dreger et al 2016) and nodal explants of fireweed (Dreger et al 2020) was reported previously. A higher regeneration rate (> 30 shoots per explants) was noted for root explants (Turker et al 2008;Dreger et al 2016), but without respect to the genotype effect.…”

“…Comparable oenothein B content (24.6 mg g −1 DW) in Ch. angustifolium shoot cultures was determined previously (Dreger et al 2020). Presence of caffeic acid pentoside is reported for the first time in in vitro cultures of fireweed, although this compound and other caffeic acid derivatives are commonly found in medicinal plants with strong antioxidant action (Tajner-Czopek et al 2020).…”

“…In addition, there are only a few works reporting in vitro culture of Ch. angustifolium (Turker et al 2008;Dreger et al 2016Dreger et al , 2020.…”

Epilobium angustifolium L. as a Potential Herbal Component of Topical Products for Skin Care and Treatment—A Review

“…The main components of EA are polyphenols, especially hydrolyzable tannins (ellagitannins), flavonoids, and phenolic acids [ 6 , 17 , 18 , 19 ]. The main phenolic acids are, among others, the following: gallic acid, 4-hydroxybenzoic acid, caffeic acid, chlorogenic acid, rosmarinic acid, ellagic acid, protocatechinic acid, and cinnamic acid [ 8 , 12 , 20 , 21 ], while the major flavonoids are the following: quercetin, myricetin, myricitrin, isomyricitrin, kaempferol, rutin, epicatechin hyperoside and isoquercetin [ 11 , 21 ]. Another important group is tannins, among others, oenothein B (OeB), which is mentioned as one of the main substances with the biological activity of this plant [ 6 , 20 , 22 ].…”

“…The main phenolic acids are, among others, the following: gallic acid, 4-hydroxybenzoic acid, caffeic acid, chlorogenic acid, rosmarinic acid, ellagic acid, protocatechinic acid, and cinnamic acid [ 8 , 12 , 20 , 21 ], while the major flavonoids are the following: quercetin, myricetin, myricitrin, isomyricitrin, kaempferol, rutin, epicatechin hyperoside and isoquercetin [ 11 , 21 ]. Another important group is tannins, among others, oenothein B (OeB), which is mentioned as one of the main substances with the biological activity of this plant [ 6 , 20 , 22 ]. EA also contains a small amount of the essential oil in leaves and stems, composed mainly of terpenes such as anethole, β-bisabolene, α-caryophyllene, and caryophyllene, eugenol, linalool, pelargol, and menthol [ 6 , 23 , 24 ].…”