Various in vitro systems of Ragged Robin (Lychnis flos-cuculi L.): a new potential source of phytoecdysteroids?

Abstract

Lychnis flos-cuculi L. is a species containing ecdysteroids, triterpenoid saponins, flavonoids, and phenolic acids, and therefore is a plant of potential medicinal value. In the presented research, diverse in vitro cultures of this taxon were developed to obtain the uniform material capable of producing ecdysteroids, including micropropagated plantlets, shoot cultures, liquid agitated whole plant cultures with fast-growing roots, and callus. A protocol of micropropagation through axillary bud formation was established using plant growth regulators at different concentrations and combinations. All the variants of plant growth regulator supplementation significantly affected a shoot proliferation rate ranging from 8 to 16 plantlets per explant, depending on the medium; DNA content of all the studied plant materials was similar. The thin-layer chromatography analysis of the extracts revealed the presence of ecdysteroids in every plant material apart from callus. The content of 20-hydroxyecdysone and polypodine B was evaluated by high-performance liquid chromatography. Agitated plantlets were in vitro cultures that efficiently formed abundant root biomass with significant concentrations of ecdysteroids. In vitro-derived adventitious roots contained twofold higher content of ecdysteroids than those of intact plants. The organs of flowering in vitro-propagated plants, transferred to experimental plot, contained twice as much ecdysteroids when compared to the organs of plants from the natural site, among which flowers were the richest in ecdysteroids. The results revealed that adventitious roots from L. flos-cuculi agitated cultures can be considered as an alternative biotechnological source of biomass rich in pharmaceutically active ecdysteroids. Key message Diverse types of Lychnis flos-cuculi in vitro cultures have been developed. HPLC analysis demonstrated much higher ecdysteroid content in cultures and regenerated plants when compared to natural site plants. Keywords Micropropagation • Callus • 20-Hydroxyecdysone • Polypodine B • Agitated plant cultures • Genome size Abbreviations 2,4-D 2,4-Dichlorophenoxyacetic acid 20HE 20-Hydroxyecdysone BAP N 6-benzylaminopurine Dic Dicamba (3,6-dichloro-2-methoxybenzoic acid) IAA Indole-3-acetic acid Kin Kinetin (N 6-furfuryladenine) MS Murashige & Skoog medium NAA 1-Naphthaleneacetic acid PGRs Plant growth regulators Pic Picloram (4-amino-3,5,6-trichloro-2-pyridinecarboxylic acid) polB Polypodine B (5β,20-dihydroxyecdysone) TDZ Thidiazuron (1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea TLC Thin-layer chromatography Communicated by Ali R. Alan.

Two Ecdysteroids Isolated from Micropropagated Lychnis flos-cuculi and the Biological Activity of Plant Material

“…The flowering herb and roots of the micropropagated plants transferred to the soil, as well as the selected callus, were characterized by phytochemical screening of 80% methanolic extracts of the plant. These results enhanced the knowledge on Ragged Robin’s chemical profile [ 6 , 18 ].…”

“…The content of ecdysteroids is almost two times higher than in intact plants: 20E present at 4.8 mg/g and polB at 3.9 mg/g in terms of the dry weight of the herb, whereas roots contain 20E at 3.7 mg/g and polB at 3.2 mg/g of dry weight. The callus rich in triterpenoid saponins, though unable to produce ecdysteroids, was obtained [ 6 , 18 ].…”

“…Until recently, little was known about the phytochemical constituents of this plant. However, it is now acknowledged that it contains phytoecdysteroids, triterpenoid saponins, flavonoids, and phenolic acids [ 1 , 2 , 3 , 4 , 5 , 6 ]. The species was introduced to in vitro cultures by our research team and the protocols of micropropagation and callus induction and development were established for the first time.…”

“…The species was introduced to in vitro cultures by our research team and the protocols of micropropagation and callus induction and development were established for the first time. Various in vitro systems, accumulating enhanced amounts of ecdysteroids, especially mature micropropagated plants, were established by Maliński et al [ 6 ].…”

“…In the plant kingdom, the most predominant ecdysteroid is 20-hydroxyecdysone (20E), while polypodine B (polB) is among the most common. Both are dominant ecdysteroids in L. flos-cuculi [ 6 ]. The adaptogenic, anabolic, and neuroprotective properties of ecdysteroids have been widely reported and reviewed [ 8 , 9 , 10 ].…”