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.