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Transgenic callus culture establishment, a tool for metabolic engineering of Rhodiola rosea L.

100%

Mirmazloum I., Forgacs I., Zok A., Pedryc A., Gyorgy Z.

Acta Scientiarum Polonorum. Hortorum Cultus

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2014

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tom 13

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nr 4

Agrobacterium tumefaciens EHA101 (pTd33) strain carrying uidA (GUS) reporter gene was used in model experiments on roseroot callus transformation. The T-DNA of pTd33 binary vector plasmid harbors nptII gene conferring resistance to kanamycin, and a uidA reporter gene, encodes the ß-glucuronidase enzyme. Roseroot seeds were sterilized and germinated on half strength MS media of which 70% germinated without any pretreatment. Calli were obtained from leaf segments of the in vitro grown seedlings. Calli was grown on solid MS medium supplemented with 1 mg l⁻¹ NAA and 0.5 mg l⁻¹ BAP. Different types of calli were obtained of which the green and compact type was chosen for transformation experiments. After co-cultivation with agrobacteria, calli were transferred to the same medium supplemented with 20 mg l⁻¹ kanamycin, 200 mg l⁻¹ carbenicillin and 300 mg l⁻¹ claforan with antioxidants (Polyclar and DTE) for selection. GUS test using a titron buffer was applied for monitoring the transformation of the calli. DNAs of 20 individual samples was extracted and subjected for PCR analysis proved the stable transformation in all of the taken samples by amplifying the nptII gene fragment. The method introduced here can be a tool for inserting and over-expressing the genes encoding for hypothesized enzymes to be involved in the biosynthesis of pharmaceutically important bioactive molecules of roseroot and therefore facilitating the applications for callus culture of roseroot in different bioreactor systems for pharmaceutical productions.

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The influence of elicitation on the tissue cultures of roseroot [Rhodiola rosea L.]

100%

Krajewska-Patan A., Mscisz A., Kedzia B., Lutomski J.

Herba Polonica

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2002

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tom 48

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nr 2

In this study the influence of the elicitation with yeast extract on the content of salidroside in the callus tissue of Rhodiola rosea was investigated. The colorimetric method for salidroside determination was used for quantitative analysis of this compound. Two series of experiments showed that the elicitor increased the salidroside content from 0.8% to 1.4% and from 0.8% to 1.5% respectively. The salidroside content in the elicited callus was twice as high as in five-year old roots of intact plants. Two types of pharmacological studies were used for the estimation of the adaptogenic activity of R. rosea extracts: determination of immunostimulating activity and determination of antistress effect.

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Effect of fertilization on roseroot (Rhodiola rosea L.) yield and content of active compounds

84%

Buchwald W., Mordalski R., Kucharski W. A., Gryszczynska A., Adamczak A.

Acta Scientiarum Polonorum. Hortorum Cultus

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2015

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tom 14

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nr 2

Roseroot (Rhodiola rosea L.) is an important medicinal plant with welldocumented adaptogenic activity. The decrease in its natural resources induces to undertake research on the introduction of this species into cultivation. The aim of the present work was to determine the effect of organic and mineral fertilization on increasing the biomass of underground parts of Rh. rosea and the level of biologically active compounds. Throughout the study period, the highest raw material yield was obtained after the application of the following doses of mineral fertilization: N – 60.0 kg·ha-1, P – 35.2 kg·ha-1, K – 83.0 kg·ha-1, without manure. The use of manure caused a decrease in the average rhizome and root weight by more than 20%. Mineral fertilization application increased the average raw material yield by about 30–40% and allowed obtaining the highest weight of underground organs of the species in question already in the fourth year of cultivation. Furthermore, the present study demonstrates that mineral fertilization does not affect substantially the level of the individual groups of compounds in the raw material, but the use of manure may significantly reduce the content of phenylpropanoids. The level of active compounds is also influenced by crop age. Two-year-old plants were characterized by the highest content of phenylpropanoids as well as by the lowest content of phenylethanoids and phenolic acids. In the following years of cultivation, no statistically significant changes were observed in the level of the analysed groups of compounds.

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Chemical investigations of biotransformed Rhodiola rosea callus tissue

84%

Krajewska-Patan A., Dreger M., Lowicka A., Gorska-Paukszta M., Mscisz A., Mielcarek S., Baraniak M., Buchwald W., Furmanowa M., Mrozikiewicz P. M.

Herba Polonica

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2007

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tom 53

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nr 4

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.

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Enhancing the biosynthesis of salidroside by biotransformation of p-tyrosol in callus culture of Rhodiola rosea L.

84%

Krajewska-Patan A., Furmanowa M., Dreger M., Gorska-Paukszta M., Lowicka A., Mscisz A., Mielcarek S., Baraniak M., Buchwalld W., Mrozikiewicz P. M.

Herba Polonica

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2007

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tom 53

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nr 1

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.

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Rhodiola rosea L. [roseroot] in vitro cultures

84%

Dreger M., Krajewska-Patan A., Furmanova M., Mscisz A., Lowicka A., Gorska-paukszta M., Mrozikiewicz P. M.

Herba Polonica

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2007

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tom 53

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nr 2

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Content of biologically active compounds in roseroot [Rhodiola sp.] raw material of different derivation

84%

Altantsetseg K., Przybyl J. L., Weglarz Z., Geszprych A.

Herba Polonica

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2007

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tom 53

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nr 4

Four populations of roseroot (Rhodiola rosea and Rh. quadrifida) originating from Poland and Mongolia were compared in respect of the content of main biologically active phenolic compounds (salidroside, p-tyrosol, trans-cinnamic alcohol, rosarin, rosavin and rosin) in the underground organs. Three solvents (75% ethanol, methanol and water) were used for the extraction of these phenolic compounds. Rh. rosea grown in Poland was characterised by similar content of salidroside and higher content of rosavin in comparison to the plants growing wild in Mongolia. The content of these compounds in Rh. quadrifida was lower than in Rh. rosea. Water appeared to be the best extraction medium for the isolation of salidroside and tyrosol. Cinnamic alcohol and its derivatives were more efficiently extracted with methanol and 75% ethanol.

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Preliminary pharmacological investigations of biotransformed roseroot [Rhodiola rosea L.] callus tissue

84%

Krajewska-Patan A., Dreger M., Lowicka A., Gorska-Paukszta M., Mscisz A., Mikolajczak P. L., Buchwald W., Furmanowa M., Mrozikiewicz P. M.

Herba Polonica

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2008

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tom 54

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nr 3

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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Effect of mother plant age and environmental factors on the yield and quality of roseroot [Rhodiola rosea L.] seeds

84%

Przybyl J., Weglarz Z., Geszprych A., Pelc M.

Herba Polonica

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2005

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tom 51

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nr 3-4

Four- and five-year-old plants gave a distinctly higher yield of seeds in comparison with six- year-old plants. Seeds originating from younger plants were also characterised by higher individual weight. Germination ability of seeds was affected both by age of mother plants and by temperature and presence of light during germination process. Higher germination ability and higher weight of seedlings was observed at substrate pH 5.5 and 6.5 in contrary to pH 3.5.

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Contents of biologically active compounds in Rhodiola rosea roots during the vegetation period

67%

Buchwald W., Mscisz A., Krajewska-Patan A., Furmanowa M., Mielcarek S., Mrozikiewicz P. M.

Herba Polonica

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2006

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tom 52

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nr 4

Studies on the contents of phenylpropanoids, salidroside, tyrosol, epigallocatechin gallate and gallic acid in Rhodiola rosea roots during the vegetation period were carried out. Rosavin was the main compound of Rhodiola rosea roots. It was found that the time of harvest of roots had a significant effect on the contents of biologically active compounds.

Ograniczanie wyników

Transgenic callus culture establishment, a tool for metabolic engineering of Rhodiola rosea L.

The influence of elicitation on the tissue cultures of roseroot [Rhodiola rosea L.]

Effect of fertilization on roseroot (Rhodiola rosea L.) yield and content of active compounds

Chemical investigations of biotransformed Rhodiola rosea callus tissue

Enhancing the biosynthesis of salidroside by biotransformation of p-tyrosol in callus culture of Rhodiola rosea L.

Rhodiola rosea L. [roseroot] in vitro cultures

Content of biologically active compounds in roseroot [Rhodiola sp.] raw material of different derivation

Preliminary pharmacological investigations of biotransformed roseroot [Rhodiola rosea L.] callus tissue

Effect of mother plant age and environmental factors on the yield and quality of roseroot [Rhodiola rosea L.] seeds

Contents of biologically active compounds in Rhodiola rosea roots during the vegetation period