enhancing the biosynthesis of salidroside by...

Vol. 53 No 1 2007

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


ANNA KRAJEWSKA-PATAN1*, MIROSŁAWA FURMANOWA2, MARIOLA DREGER1, MAŁGORZATA GÓRSKA-PAUKSZTA1, ANNA ŁOWICKA1, ALINA MŚCISZ1, SEBASTIAN MIELCAREK1, MAREK BARANIAK1, WALDEMAR BUCHWALD1, PRZEMYSŁAW M. MROZIKIEWICZ1

1Research Institute of Medicinal Plants, Libelta 27, 61-707 Poznań, Poland2Department of Biology and Pharmaceutical Botany, Medical University, Banacha 1, 02-097 Warsaw, Poland

*corresponding author: tel.: +4861 6659540 e-mail: [email protected]

S u m m a r y

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.

Key words: Rhodiola rosea L.,callus tissue, biosynthesis, salidroside

INTRODUCTION

Rhodiola rosea L., roseroot (synonyms: Golden root, Arctic root) is a herbaceous perennial plant belonging to Crassulaceae family. Roseroot grows in Arctic and in mountain regions of Asia, North America and Europe. This plant is attributed to many physiological and pharmacological activities: stimulating the central nervous

A. Krajewska-Patan, M. Furmanowa, M. Dreger, M. Górska-Paukszta, A. Łowicka, A. Mścisz, S. Mielcarek, .

system (CNS) [1-3], enhances physical and mental work performance [4-7], elimina-tes fatigue and possess adaptogenic [8-12], cardioprotective [13], anticancer [14, 15], antioxidant [16-19] and antimicrobial activity [20, 21]. Some activities of Rhodio-la rosea extracts have been proved in pharmacological and clinical studies [5-7].

The roots of Rhodiola rosea L. contain a wide range of biologically active compo-unds: phenylpropanoids – rosavin, rosarin, rosin (compounds specific for R. rosea L.) [22-26], phenolic compounds – salidroside, tyrosol [17, 24, 26, 27], flavonoids – rodionin, rodiolin, rhodiosin, acetylrodalgin and tricin [28-30], phenolic acids – gallic acid, chlorogenic acid, hydroxycinnamic acid [27,31], monoterpenes [31], β-sitosterol, daukosterol [32], tannins [31], fatty acids [33], cyanogenic glucosi-de – lotaustralin [34] as well as essential oils – n-decanol and geraniol [35]. The variability of chemical composition depends on the year of the crops, climatic conditions and the plants origin [36].

Salidroside, tyrosol and phenylopropanoids (rosavins) are supposed to be re-sponsible for the CNS-stimulating and adaptogenic properties. The most inte-resting feature of the plant is the psychostimulating activity, which arouse the growing interest of pharmaceutical industry. The biomass obtained with the use of biotechnological methods might be an alternative way of producing valuable plant raw material, independent from climatic conditions.

The biotechnological investigations on Rhodiola rosea have been carried out in Russia, Poland, Finland, Germany and others. The biotransformation is one of the most effective methods of enhancing the content of biologically active sub-stances. There are several publications confirming that the addition of the precur-sors of biosynthesis (cinnamyl alcohol, p-tyrosol) might significantly increase the content of rosavins or salidroside in callus tissue of R. rosea [37-45]. As a result, the content of salidroside in biomass treated by p-tyrosol was higher that in roots of in vivo plants [38, 40, 42, 44, 45].

The main goal of this research was to enhance the content of salidroside by exogenous addition of p-tyrosol in tissue cultures and determine the most effec-tive concentration of the precursor.

MATERIAL AND METHODS

The callus tissue cultured on solid medium and compact callus aggregate (CCA) were used in the experiments. The callus was obtained from hypocotyls of the seedlings and cultured on Murashige-Skoog (MS) medium [46] supplemented with α-naphtaleneacetic acid (NAA), benzyladenine (BA) and adenine chloride (solid media and CCA).

The suspension culture of CCA was initiated from callus cultured on solid me-dia. The callus clumps (about 20 g, age of culture: 4 weeks) were transferred into 50 ml of liquid medium (MS supplemented with NAA, BA and adenine chloride) of 250 ml flask and shaken at 110 rpm. The suspension culture had been subcultured every 14 days.

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The exogenous p-tyrosol (Sigma) was added to callus culture to enhance the production of salidroside via biotransformation. The sterile filtred p-tyrosol (in the methanol solution) was added to medium at concentration of 2.5 mM/L (only into liquid medium) and 5 mM/L (both into liquid and solid medium) in the day of the inoculation.

The callus was collected and dried (25oC) on the 7th day following the biotrans-formation. The fresh and dried biomasses were measured. The contents of p-tyro-sol and salidroside were determined in callus tissue after biotransformation and in callus with no addition of precursor as well as in the roots of plants cultured in vivo.

CHROMATOGRAPHICAL PROCEDURES

In this study HPTLC, HPLC and spectrophotometric methods were used for che-mical analyses. The hydroalcoholic extracts from roots and callus tissue were pre-pared. The presence and content of salidroside and tyrosol was determined in cal-lus treated with p-tyrosol and in the control tissue (without addition of p-tyrosol). For HPLC analysis, samples (1 g each) were extracted with 20 ml of 70% methanol at boiling point for 15 min. Extraction was repeated three times with 70% aqueous methanol. HPLC analysis was performed on Agilent 1100 HPLC system, equipped with photodiode array detector. For all separations the Lichrospher 100 RP 18 co-lumn (250x4 mm, Merck) was used. The mobile phase consisted of 0.2% phosphoric acid in water (A) and acetonitrile (B), applied in the gradient elution (see Table 1).

Ta b l e 1 .

Scheme of gradient elution.

time [min] 0.2% H3PO4 aq [%] CH3CN [%]0.00 95.0 5.030.00 80.0 20.035.00 80.0 20.040.00 20.0 80.056.00 20.0 80.060.00 95.0 5.070.00 95.0 5.0

The following rate adjusted to 1 ml/min., the wavelength detection set to DAD at λ=205.5 nm, 220.4 nm, 254.4 nm. Then 20 μL of samples was injected. All separations were performed at the temperature of 24oC. Peaks were assigned by spiking the samples with standard compounds and comparison of the UV-spectra and retention times.

A. Krajewska-Patan, M. Furmanowa, M. Dreger, M. Górska-Paukszta, A. Łowicka, A. Mścisz, S. Mielcarek,

RESULTS

In the first 4 weeks after transferring the callus into liquid medium the culture of compact callus aggregate was established (Fig. 1 and 2). The size of the formed aggregates amounted approximately to 0.5–1.0 cm.

Fig 1. Rhodiola rosea CCA

Fig. 2. R. rosea callus on a solid medium.

The application of 5 mM tyrosol resulted in the increase of tyrosol and salidro-side content in callus cultured both on solid and in the liquid media. After 7 days of biotransformation the content of p-tyrosol in callus approximated: 5.76 mg/g

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on solid medium and 8.16 mg/g CCA (Fig. 3 and 4). The yield of p-tyrosol was 0.6% (in callus on solid medium) and 0.8% (in CCA). Salidroside content in transformed tissue (after 7 days) was 23.15 mg/g (on solid medium) and 43.22 mg/g (CCA) (Fig. 3, 4, 6, 7). The yield of salidroside was approximately 3.1% (solid medium) and 4.3% (CCA).

Fig. 3. The content of salidroside and tyrosol in R. rosea callus cultured on solid medium after the application of 5 mM tyrosol.

Fig. 4. The content of salidroside and tyrosol in R. rosea CCA after the application of 5 mM tyrosol.

The addition of 2.5 mM tyrosol into CCA culture induced the increase of the content of salidroside to 34.73 mg/g and tyrosol to 1.80 mg/g (Fig. 5, 8, 9). The 3.5% yield of salidroside was obtained.


Fig. 5. The content of the salidroside and tyrosol in R. rosea CCA after the application of 2.5 mM tyrosol.

The adverse effect was observed in biomass. The set back of biomass CCA was 20% (tyrosol concentration 5 mM) and 9% (at 2.5 mM of tyrosol), compared to control.

Neither salidroside nor tyrosol were released into the medium.

DISCUSSION

The use of CCA instead of suspension cells as the efficient tool for biotransfor-mation in Rhodiola sp. cultures was described by Xu et al., Wu et al. and György et al. [41, 42, 47-49].

The obtained contents of salidroside both in CCA and in culture on solid me-dium at both precursor concentrations (2.5 and 5 mM) are higher than salidroside content in roots of plants in vivo (2%) and comparable with another studies [38, 40, 44, 49]. In the presented investigation the biotransformation of CCA with use of 5 mM tyrosol resulted in the highest content of salidroside: 43.22 mg/g (4.3%).

Up to now the highest content of intracellular salidroside – 154.95 mg/g (15.4%) – was detected by Xu et al. [50] as a result of repeated 3 mM addition of tyrosol into R. sachalinensis suspension culture. Xu et al. observed that the addition of tyrosol at the concentration higher than 3mM caused a decrease of the salidroside content and was toxic for the cells of suspension culture. So high salidroside content and toxic effect has not been observed neither by any authors nor in this study.

The obtained salidroside content (43.22 mg/g (4.3%)) is comparable to results described by Wu et al. [49] who received 57.72 mg/g (5.77%) of salidroside as the effect of 4 mM tyrosol application into R. sachalinensis compact callus aggregates.

The biotransformation of R. rosea suspension culture by glucosylation of tyrosol was studied by Furmanowa et al. [38, 40]. The exogenous addition of 2.5 mM tyro-sol enhanced salidroside yields to 1.8 and to 2.3%. The same concentration of ty-rosol in the presented experiment induced the increase of salidroside production

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to 3.5% (CCA). Similar study carried out by György at al. [44] resulted in salidroside yields: 2.62% and 2.72% with use of 2 and 3 mM tyrosol, respectively.

The decrease of biomass growth was observed in most of the studies [44, 49, 50]. The biomass decrease was proportionally dependent on the tyrosol concen-tration but its value was different: from 10% up to one third of control [44, 50]. In the presented studies the decrease of biomass growth was proportionally depen-dent on tyrosol concentration and varied from 10% to 20% (at 2.5 mM and 5 mM tyrosol concentration, respectively).

The attempt of R. rosea culture biotransformation on solid medium was per-formed. The production of salidroside enhanced 23.15 mg/g (3.1%). This is a rela-tively high value considering that the availability of precursor in a solid medium is incomparable to a liquid medium.

The obtained results confirmed that even on a solid medium the significant improvement of salidroside production in R rosea cultures is possible.

ACKNOWLEDGEMENTS

The work was supported with a grant by the Polish Comunitte for Scientific Research No 2P05F03228

REFERENCES

1. Salnik BU. Effect on several stimulators on central nervous system energy metabolism during muscular workload [dissertation]. Tomsk, Russia. Tomsk State Medical Institute 1970.

2. Saratikov A, Marina TF, Fisanova LL. Effect of golden root extract on processes of serotonin synthesis in CNS. J Biol Sci 1978; 6:142.

3. Krajewska-Patan A, Mikołajczak PŁ, Okulicz-Kozaryn I, Bobkiewicz-Kozłowska T, Buchwald W, Łowicka A, Furmanowa M, Dreger M, Górska-Paukszta M, Mścisz A, Mrozikiewicz PM. Rhodiola rosea extracts from roots and callus tissues – study on relationship between their chemical contents and CNS affecting pharmacological activity. 11th International Congress of Polish Herbal Committee. Poznań, 24-25 June2005. Herba Pol 2005; 51(Suppl.1):105-6.

4. Lazarova MB, Markovska VL, Petkov VV, Mosharrof A. Effects of meclofenoxate and extr. Rhodiola rosea L. on electroconvulsive shock - impaired learning and memory in rats. Methods Find Exp Clin Pharmacol 1986; 8(9):547-52.

5. Darbinyan V, Kteyan A, Panossian A et al. Rhodiola rosea in stress induced fatigue: a double blind cross - over study of a standarized extract SHR-5 with a repeated low - dose regiment on the mental performance of healthy physicians during night duty. Phytomedicine 2000; 7(5):365-71.

6. Spasov AA, Wilkman GK, Mandrikov VB et al. A double-blind, placebo-controlled pilot study of the stimulating and adaptogenic effect of Rhodiola rosea SHR-5 extract on the fatigue of students caused by stress during an examination period with a repeated low-dose regimen. Phytomedicine 2000; 7(2):85-9.

7. Spasow AA, Mandrikov VB, Mironova IA. The effect of preparation rhodiosin on the psychophysiological and physical adaptation of students to an academic load. Eksp Klin Farmakol 2000; 63(1):76-78.

8. Panossian A, Wikman G, Wagner H. Plant adaptogens. III. Earlier and more recent aspects and concept on their mode of action. Phytomedicine 1999; 6(4):287-300.

9. Boon-Niermeijer EK, van der Berg A, Wikman G, Wiegant FA. Phyto-adaptogens protect against environmental stress-induced death of embryos from freshwater snail Lymnaea stagnalis. Phytomedicine 2000; 7(5):389-99.

10. Kelly GS. Rhodiola rosea: a possible plant adaptogen. Altern Med Rev 2001; 6(3):293.


11. Panossian A, Wagner H. Stimulating effect of adaptogens: an overview with particular reference to their efficacy following single dose administration. Phytother Res 2005; 19(10):819-38.

12. Kelly GS. Rhodiola rosea: a possible plant adaptogen. Altern Med Rev 2001; 6(3):293.13. Maslova LV, Kondratev BIu, Maslov LN et al. The cardioprotective and antiadrenergic activity of an

extract of Rhodiola rosea in stress. Eksp Klin Farmakol 1994; 57(6):61-63.14. Udintsev SN, Shakhov VP. Decrease of cyclophosphamide haematotoxity by Rhodiola rosea root eextract

in mice with Ehrlich and Lewis transplantable tumors. Eur J Cancer 1991; 27:1182.15. Udintsev SN, Krylova SG, Fomina TI. The enhancement of the efficacy of adriamycin by using

hepatoprotectors of plant origin in metastases of Ehrlich’s adenocarciroma to the liver in mice. Vopr Onkol 1992; 38:1217-22

16. Furmanowa M, Skopińska-Różewska E, Rogala E, Hartwich M. Rhodiola rosea L. in vitro culture – phytochemical analysis and antioxidant action. Acta Soc Bot Pol 1998; 67:69.

17. Furmanowa M, Kędzia B, Hartwich M, Kozłowski J, Krajewska-Patan A, Mścisz A, Jankowiak J. Phytochemical and pharmacological properties of Rhodiola rosea L. Herba Pol 1999; 45:108-13.

18. Battistelli M, De Sanctis R, De Bellis R, Cucchiarini L, Dacha M, Gobbi P. Rhodiola rosea as antioxidant in red blood cells: ultrastructural and hemolytic behaviour. Eur J Histochem 2005; 49(3):243-54.

19. De Sanctis R, De Bellis R, Scesa C, Mancini U, Cucchiarini L, Dacha M. In vitro protective effect of Rhodiola rosea extract against hypochlorous acid-induced oxidative damage in human erythrocytes. Biofactors 2004; 20(3):147-59.

20. Furmanowa M, Starościak B, Lutomski J, Kozłowski J, Urbańska N, Krajewska-Patan A, Pietrosiuk A, Szypuła W. Antimicrobial effect of Rhodiola rosea L. roots and callus extracts on some strains of Staphylococcus aureus. Herba Pol 2002; 48:23.

21. Krajewska-Patan A, Kędzia B, Dreger M, Mścisz A, Buchwald W, Furmanowa M, Mrozikiewicz PM. Antimicrobial activity of Rhodiola rosea extracts. Proceeding of 7th Congress of the European Association for Clinical Pharmacology and Therapeutics. 25-29 June2005, Poznań, Poland. Abstract Book. Basic Clin Pharmacol Toxicol 2005; 97(Supp.1):38.

22. Zapesochnaya G, Kurkin V. Cinnamic glycosides of Rhodiola rosea rhizomes. Khim Prirod Soed 1982; 6:723.23. Kir’yanov A, Bondarenko L, Kurkin V, Zapesochnaya G. Chromatospectrophotometric assay for measuring

rosavidin in Rhodiola rosea rhizomes. Khim-Farm Zh 1988; 22(4):451.24. Kir’yanov A, Bondarenko L, Kurkin V, Zapesochnaya G et al. Determination of biologically active

consistuents of Rhodiola rosea rhizomes. Kim-Prir Soedin 1991; 3:320.25. Kurkin V, Zapesochnaya G, Shchavlinskii A et al. Russ. Otkrytiia, Izobretaniia 1985:27,30 Patent.26. Saratikov AS, Krasnov EA, Khnykina LA et al. Rhodioloside, a new glicoside from Rhodiola rosea and its

pharmacological properties. Pharmazie 1970; 23(7):392-5.27. Dubichev A, Kurkin W, Zapesochnaya G, Vorontsov V. HPLC study of Rhodiola rosea rhizomes. Khim Prir

Soedin 1991; 2:188.28. Kurkin V, Zapesochnaya G, Klyaznika V. Rhodiola rosea rhizome flavonoids. Khim Prir Soedin 1982; 5:581.29. Kurkin V, Zapesochnaya G, Shchavlinskii A. Flavonoids of rhizomes of Rhodiola rosea. III. Khim Prir

Soedin 1984; 5:657.30. Zapesochnaya G, Kurkin V. Flavonoids of Rhodiola rosea rhizomes. I. Khim Prirod Soed 1983; 1:23.31. Kurkin V, Zapesochnaya G. Chemical composition and pharmacological properties of Rhodiola sp. Plants

Review. Khim-Farm Zh 1986; 20(10):1231.32. Kurkin V, Zapesochnaya GG, Kir’yanov AA et al. Quality of raw Rhodiola rosea L. material. Khim-Farm Zh

1989; 23:11.33. Anilakumar, Pooja KR, Khanum, Farhath, Bawa AS. Phytoconstituents and antioxidant potency of

Rhodiola rosea - a versatile adaptogen. J Food Biochem 2006; 30(2):203-214(12).34. Akgul Y, Ferreira D, Abourashed EA, Khan IA. Lotaustralin from Rhodiola rosea roots. Fitoterapia 2004;

75(6):612-14.35. Rohloff J. Volatiles from rhizomes of Rhodiola rosea L. Phytochemistry 2002; 59(655):61.36. Buchwald W, Mścisz A, Krajewska-Patan A, Furmanowa M, Mielcarek S, Mrozikiewicz PM. Contents of biological

active compounds in Rhodiola rosea roots during the vegetation period. Herba Pol 2006; 52(4):39-43.37. Michalska M, Alfermann AW, Furmanowa M. Rosavins as a product of glucosylation by Rhodiola rosea

Cell Suspension Cultures. 45Th Annual Congress of the Society for Medicinal Plant Research. Regensburg (Germany), 7–12 September 1997. Poster L 12.

...

Vol. 53 No 1 2007


38. Furmanowa M, Hartwich M, Alferman AW. Salidroside as a product of biotransformation by Rhodiola rosea cell suspension cultures. 2000 years of natural product research. 26-30 July 1999 (Holland), Book of Abstracts, p.152.

39. Furmanowa M, Hartwich M, Alferman AW, Koźmiński W, Olejnik M. Rosavins as a product of glucosylation by Rhodiola rosea (roseroot) cell cultures. Plant Cell Tissue Organ Cult 1999; 56(105):110.

40. Furmanowa M, Hartwich M, Alferman AW. Glucosylation of p-tyrosol to salidroside by Rhodiola rosea L. cell cultures. Herba Pol 2002; 48(2):71-6.

41. György Z, Tolonen A, Pakonen M, Neubauer P, Hohtola A. Enhancing the production of cinamyl glucosides in compact callus aggregate cultures of Rhodiola rosea by biotransformation of cinnamyl alcohol. Plant Sci 2004, 166(1):229-36.

42. Tolonen A, György Z, Pakonen M, Neubauer P, Hohtola A. LC/MS/MS Identyfication of glucosides produced by biotransformation of cinnamyl alcohol in Rhodiola rosea compact callus aggregates. Biomed Chromatogr 2004, 18:550-8.

43. György Z, Tolonen A, Neubauer P, Hohtola A. Enhanced biotransformation capacity of Rhodiola rosea callus cultures for glycoside production. Plant Cell Tissue Organ Cult 2005; 83:129-35.

44. György Z. Glucoside production by in vitro Rhodiola rosea cultures. Acta Universitatis Ouluensis C Technica 2006:244.

45. Krajewska-Patan A, Mrozikiewicz PM, Mścisz A, Dreger M, Górska-Paukszta M, Łowicka A, Buchwald W, Mielcarek S. Biotransformation of p-tyrosol by callus tissue of Rhodiola rosea L. 5th International of Symposium on Chromatography of Natural Products. Lublin (Poland), -22 June 2006. Books of Abstracts:135.

46. Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tabacco cultures. Physiol Plant 1962; 15:473.

47. Xu Jianfeng, Su Zhiguo, Feng Pusun. Suspension culture of compact callus aggregates of Rhodiola sachalinensis for improved salidrosise production. Enzyme Microb Technol 1998; 23:20-7.

48. Xu JF, Ying PQ, Han AM, Su ZG. Enhanced salidroside production in liquid-cultivated compact callus aggregates of Rhodiola sachalinensis: manipulation of plant growth regulators and sucrose. Plant Cell Tissue Organ Cult 1998, 55(1):53-8.

49. Shuangxiu Wu, Yuangang Zu, Madeline Wu. High yield production of salidroside in the suspension culture of Rhodiola rosea. J Biotechnol 2003; 106:33-43.

50. Xu JF, Su ZG, Feng PS. Activity of tyrosol glucosyltransferase and improved salidroside production through biotransformation of tyrosol in Rhodiola sachalinensis cell culture. J Biotechnol 1998; 61(1):69-73.

51. Xu JF, Liu CB, Han AM, Feng PS, Su ZG. Strategies for the improvement of salidroside production in cell suspension cultures of Rhodiola sachalinensis. Plant Cell Reports 1998; 17(4):288-93.

WZMOŻONA BIOSYNTEZA SALIDROZYDU W WYNIKU BIOTRANSFORMACJI P-TYROZOLU W KULTURACH KALUSOWYCH RÓŻEŃCA GÓRSKIEGO RHODIOLA ROSEA L.

ANNA KRAJEWSKA-PATAN1*, MIROSŁAWA FURMANOWA2, MARIOLA DREGER1, MAŁGORZATA GÓRSKA-PAUKSZTA1, ANNA ŁOWICKA1, ALINA MŚCISZ1, SEBASTIAN MIELCAREK1, MAREK BARANIAK1, WALDEMAR BUCHWALD1, PRZEMYSŁAW M. MROZIKIEWICZ1

1 Instytut Roślin i Przetworów Zielarskich, ul. Libelta 27, 61-707 Poznań2 Katedra i Zakład Biologii i Botaniki Farmaceutycznej Akademia Medyczna ul. Banacha 1, 02-097 Warszawa

*autor, do którego należy kierować korespondencję: +4861 6659540, e-mail: [email protected]


S t r e s z c z e n i e

Celem badań było zwiększenie zawartości salidrozydu w kulturach kalusowych różeńca górskiego poprzez dodanie p-tyrozolu bezpośrednio do pożywki. Eksperyment obejmował zarówno kultury hodowane na pożywce stałej (wg Murashiga i Skooga z dodatkiem NAA, BA i chlorku adeniny), jak i na pożywce płynnej (CCA). Tyrozol dodawano w stężeniu 5 mM/L i 2,5 mM/L (tylko w przypadku CCA) w dniu inokulacji. Po 7 dniach od dodania 5 mM ty-rozolu zawartość salidrozydu wynosiła: 23,15 mg/g (kalus na stałej pożywce) i 43,22 mg/g (CCA). Zastosowanie 2,5 mM/L pozwoliło zwiększyć biosyntezę salidrozydu do zawartości 34.73 mg/g w CCA. Zaobserwowano również zmniejszony przyrost biomasy w stosunku do kontroli. Nie stwierdzono wydzielania salidrozydu do pożywki.

Słowa kluczowe: Rhodiola rosea, właściwości adaptogenne, kultury in vitro, biotransformacja

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enhancing the biosynthesis of salidroside by...

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