Accumulation of rosmarinic, chlorogenic and caffeic acids in in vitro cultures of Eryngium planum L.

Eryngium planum L. cell and organ cultures were maintained on Murashige and Skoog media (MS), supplemented with exogenous hormones of different types and various concentrations for high biomass growth. The callus and cell suspension cultures were treated with increased sucrose concentration and/or elicited by methyl jasmonate for the enhancement of selected phenolic acids accumulation. Three phenolic acids, rosmarinic acid (RA), chlorogenic acid (CGA) and caffeic acid (CA), were detected by HPLC-DAD in those cultures. The sum of their content in the dry material was found to be higher in the shoot culture (3.95 mg g^?1), root culture (7.05 mg g^?1), callus (6.20 mg g^?1) and cell suspension (2.04 mg g^?1) than in the leaves (1.87 mg g^?1) and roots (0.76 mg g^?1) of intact plants. The major compound of in vitro cultures was always rosmarinic acid. The content of RA could be increased approximately threefold (16.24 mg g^?1) in the callus culture and approximately twofold (3.91 mg g^?1) in the cell suspension culture by elicitation with 100 μM methyl jasmonate (MeJA). The higher concentration of sucrose (S) in the medium (5, 6 %) led to over a twofold increase of CGA content in the callus culture (2.54 mg g^?1). The three mentioned phenolic acids have been found in E. planum undifferentiated and differentiated in vitro cultures for the first time.     

The Effect of Nutritional Factors and Plant Growth Regulators on Micropropagation and Production of Phenolic Acids and Saponins from Plantlets and Adventitious Root Cultures of Eryngium maritimum L.

Eryngium maritimum L. is a valuable medicinal species, but since it is protected plant, collection from natural populations is forbidden. Therefore, establishing an efficient system for micropropagation of this species is desirable. To determine the optimal nutritional factors needed for shoot multiplication, root development and secondary metabolites accumulation, different media and plant growth regulators were tested. The highest plant regeneration efficiency (over 96 %), with 4.4 shoots per explant was induced on Murashige and Skoog (MS) medium supplemented with 1.0 mg L^?1 benzyladenine (BA) and 0.1 mg L^?1 indole-3-acetic acid (IAA). The in vitro-regenerated shoots were rooted (83.3–100 %) and transferred to an experimental plot with 62 % efficiency. Flow cytometric analysis revealed no variation in nuclear DNA content in field- and in vitro-delivered plant material. Ultra high performance liquid chromatography (UHPLC) indicated that multiple shoots and roots from in vitro-regenerated plantlets and adventitious root cultures maintained the production of rosmarinic (RA) and chlorogenic (CGA) acids and triterpenoid saponins found in the rosette leaves and roots of E. maritimum intact plants. UHPLC revealed a 12-fold increase of RA and CGA and 3.2-fold higher accumulation of triterpenoid saponins in roots of in vitro-derived plantlets in comparison to roots from field-grown plants. Adventitious root cultures allowed continuous growth of excised root in liquid media with or without exogenous auxins. The roots grown in liquid medium supplemented with 0.1 mg L^?1 IAA showed higher (227-fold) phenolic acids accumulation than those without auxin. Obtained results confirmed that micropropagation is a useful strategy in the protection of endangered species and a renewable source of a high quality plant material for secondary metabolites production.     

Phenolic acid and DNA contents of micropropagated Eryngium planum L.

A protocol for in vitro production of genetically uniform populations of the medicinal plant Eryngium planum, rich in selected phenolic acids, has been established. Shoot-tips were collected from axenic seedlings and grown on a Murashige and Skoog basal medium supplemented with 6-Benzyladenine (BA) and Indole-3-acetic acid (IAA). The highest shoot proliferation efficiency (17 shoots per explant) was obtained when 1.0 mg L^?1 BA and 0.1 mg L^?1 were added. Proliferating shoots were rooted and transferred to soil (89 % frequency of survival). Flow cytometric analysis of intact (field-grown) and microrpropagated plants revealed that all plants were uniform in genome size and had similar DNA contents. Thin-layer chromatography (TLC) analysis indicated that multiple shoots and roots from in vitro-derived plants produced high amounts of phenolic acids, primarily of rosmarinic acid (RA). Levels of phenolic acids in in vitro-derived plants were similar to those of intact plants. Furthermore, high-performance liquid chromatography revealed that root cultures in liquid medium accumulated substantial levels of RA. Thus, rapid establishment of in vitro-grown organ cultures of E. planum can also serve as reliable sources for bioactive compounds.     

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

Plant Cell, Tissue and Organ Culture (PCTOC) - Lychnis flos-cuculi L. is a species containing ecdysteroids, triterpenoid saponins, flavonoids, and phenolic acids, and therefore is a plant of...     

Comparative analysis of phenolic acids and flavonoids in shoot cultures of Eryngium alpinum L.: an endangered and protected species with medicinal value

Plant Cell, Tissue and Organ Culture (PCTOC) - Phenolic acids and flavonoids, important bioactive compounds of polyphenols, play a significant role in plants; their impact, mainly as antioxidants,...