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11 polysaccharide fractions were isolated from the Echinacea purpurea herb (collected from the cultivation) and 8 compounds were isolated from the callus tissue from in vitro cultures. The HPLC analysis of hydrolysed polysaccharides indicated a great heterogeneity of the herb fractions. They differ significantly from the compounds obtained from the callus tissue, which are mostly arabinogalactans with an addition of xylose.
Several species of Rhodiola genus (Crassulaceae family) like Rhodiola kirilowii and Rhodiola rosea are used in official or traditional medicine. The aim of this study was to determine qualitative and quantitative content of proanthocyanidins using ultra performance liquid chromatograph connected to a tandem mass spectrometer (UPLC MS/MS method) in the callus tissues and in the transformed roots (infected by Agrobacterium rhizogenes LBA 9402 strain) of R. kirilowii and R. rosea. This validated assay allows to determine the content of five flavan-3-ols: (+)-catechin, (-)-epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate (ECG), (-)-epigallocatechin gallate (EGCG). Our results concerning the material from in vitro cultivation of R. kirilowii and R. rosea indicate that R. rosea callus can be a better source of catechin when compared with other tested materials, especially when the content of (-)-gallate epigallocatechin is taken under consideration (3.429 mg/100 g of dry powdered material). The application of UPLC MS/MS method allowed to determine the content of proanthocyanidins in tested samples with satisfactory precision and can be used in the phytochemical investigations of Rhodiola sp. in vitro cultivated tissues.
The main aim of this study was to search the influence of exogenous addition of rosavin precursor: cinnamyl alcohol on the enhancing of rosavins content in callus culture of R. rosea, cultured on solid and liquid media (CCA). This is the first report – according to available literature – which concerns its biotransformation on solid medium conditions. The two strains of R. rosea tissue cultures showed different ability of cinnamyl alcohol glycosides production: both of them produced rosin (with or without supplementation), but the obtained level of rosavin production was notable higher in case of supplementation of the strain induced from axially buds of R. rosea. The exogenous cinnamyl alcohol was added into medium at concentration of 2.5 mM/L or 5 mM/L in the day of the inoculation. The application of 2.5 mM cinnamyl alcohol resulted in the increase of rosin content in the callus started from hypocotyle to very high levels: 1056.183 mg/100 g on solid medium and 776.330 mg/100 g in liquid medium. The content of rosavin showed the same growing tendency, but the final concentration of this phenylopropanoid in the supplemented callus tissue was about 7 times lower as compared to the roots of intact plant (63.603 mg/100 g). Addition of cinnamyl alcohol also enhanced rosarin biosynthesis but only in small amount: to 4.896 mg/100 g on solid medium. Callus tissue obtained from axially buds and treated by cinnamyl alcohol (2.5 mM) produced rosavin in a higher concentration: 92.801 mg/100 g and reached one fifth part of the amount produced by roots. The process of supplementation with cinnamyl alcohol influenced the enhanced biosynthesis of another bioactive substances as well (salidroside, tyrosol, chlorogenic acid). The obtained results confirmed that even on a solid medium the callus tissue can produce the characteristic active substances and the concentration of some of them, mainly rosin and rosavin, can be significantly improved by addition of the precursors to the medium.
The aim of this research was to enhance the content of salidroside by exogenous addition of p-tyrosol in R. rosea tissue cultures. The callus tissue cultured on solid medium (MS with addition of NAA, BAP and adenine chloride) and compact callus agregate (CCA) were used in the experiments. The p-tyrosol was added to medium at a concentration of 5 mM/L (both into liquid and solid medium) and at concentration of 2.5 mM/L (only into liquid medium) in the day of the inoculation. The content of salidroside approximated: 23.15 mg/g (on solid medium) and 43.22 mg/g (CCA) after 7 days of 5 mM/L p-tyrosol application. The yield of salidroside was 3.1% (solid medium) and 4.3% (CCA). The addition of 2.5 mM tyrosol to CCA culture induced the increase of the content of salidroside to 34.73 mg/g and 3.5% yield of salidroside was obtained. The adverse effect was observed in biomass. Salidroside was not released into the medium.
Callus tissues from Rhodiola rosea L., a plant popular in traditional Russian and Asian medicine, treated with biosynthesis precursors of salidroside (p-tyrosol) and rosavins (cinnamic alcohol), were tested on the animals for the pharmacological activities of the extracts. Authors investigated the behavioural activity using locomotor activity and the motor coordination test. A single administration of extract from the callus tissue non treated with precursors resulted in stimulant effects on rats. The extract enriched in rosavins (after biotransformation) showed the inhibition of activity of tested animals. The more detailed studies concerning the exact nature of the effects especially after chronic administration are needed.
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