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 vitroderived 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.

ABSTRACT:In vitro shoot culture of four Solidago species, S. virgaurea L., S. canadensis L., S. gigantea Ait. and S. graminifolia (L.) Salisb. were established from aseptically germinated seedlings. Proliferated axillary shoots were multiplied rapidly on Murashige and Skoog's medium, supplemented with 9.3 µM kinetin and 11.4 µM indole-3-acetic acid. Rooted plantlets were transferred to the soil, then grown in the field. Aerial parts were collected from clonalpropagated Solidago species during the flowering stage. The essential oils, obtained from air-dried plant material by hydrodistillation, were studied by GC and GC-MS. Slight differences were found in the yield and composition of essential oils between the micropropagated plants and wild plants growing in natural sites.

No abstract

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 lM 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.

Ecdysteroids are secondary metabolites, widely distributed in the animal and plant kingdoms. They have a wide range of pharmacological effects in vertebrates, including mammals, most of which are beneficial for humans. Therefore, they have become compounds of interest for the pharmaceutical industry due to their adaptogenic, anabolic, hypoglycaemic, hypocholesterolaemic and antimicrobial activities, which are still being researched. Nowadays, ecdysteroids are present as active ingredients in bodybuilding supplements. Because of their complex structures, their chemical synthesis seems unprofitable and impractical. Due to high content of ecdysteroids in many plants, they are primarily obtained by extraction of the plant material. Plant in vitro cultures provide an alternative source of these compounds, helping to avoid problems associated with field production—such as variable yield or dependence on environmental factors, as well as limited availability of natural resources. Plant cell and tissue cultures may be suggested as alternatives for the production of plant biomass rich in pharmaceutically active ecdysteroids. Moreover, the use of common biotechnological strategies, such as elicitation or precursor feeding, may further increase the yield and improve production of these compounds. In this paper, we describe general information about ecdysteroids: their structure, biosynthesis, distribution, role in plants, and we review recent studies on micropropagation of ecdysteroid-producing plants and cell cultures, and potential ability of ecdysteroids enhancement in in vitro cultures.