Abstract
The common effect of NO3 −, KH2PO4 and sucrose on the biosynthesis of hyoscyamine by diploid and tetraploid Datura stramonium L. hairy root cultures were studied. Polynominal regression models were elaborated for the description of hyoscyamine biosynthesis as a consequence of variation of the investigated variables (NO3 − between 31 and 47 mM, and sucrose between 20.00 and 40.00 g/l for diploid and NO3 − between 31 and 47 mM; KH2PO4 between 0.50 and 1.25 mM, and sucrose between 50 and 70 g/l, for tetraploid hairy roots, respectively). Optimization procedures allowed us to establish the optimal concentrations of the investigated variables and to propose the modified MS nutrient media, with 1.10 g/l KNO3, 0.17 g/l KH2PO4, and 40 g/l sucrose, for diploid and with 1.10 g/l KNO3, 0.17 g/l KH2PO4, and 50 g/l sucrose, for tetraploid Datura stramonium L. hairy root cultures. The proposed modified MS media provided optimal hyoscyamine production yield for both investigated hairy root cultures.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baiza AM, Quiroz-Moreno A, Ruiz JA, Loyola-Vargas VM (1999) Genetic stability of hairy root cultures of Datura stramonium. Plant Cell Tissue Organ Cult 59(1):9–17
Bensaddek L, Gillet F, Saucedo JEN, Fliniaux MA (2001) The effect of nitrate and ammonium concentrations on growth and alkaloid accumulation of Atropa belladonna hairy roots. J Biotechnol 85(1):35–40
Berkov S, Pavlov A (2004) A rapid densitometric method for the analysis of hyoscyamine and scopolamine in solanaceous plants and their transformed root cultures. Phytochem Anal 15(3):141–145
Berkov S, Philipov S (2002) Alkaloid production in diploid and autotetraploid plants of Datura stramonium. Pharm Biol 40(8):617–621
Berkov S, Pavlov A, Kovatcheva P, Stanimirova P, Philipov S (2003) Alkaloid spectrum in diploid and tetraploid hairy root cultures of Datura stramonium. Z Naturforsch [C] 58(1–2):42–46
Boitel-Conti M, Laberche JC, Lanoue A, Ducrocq C, Sangwan-Norreel BS (2000) Influence of feeding precursors on tropane alkaloid production during an abiotic stress in Datura innoxia transformed roots. Plant Cell Tissue Organ Cult 60(2):131–137
De Jesus-Gonzalez L, Weathers PJ (2003) Tetraploid Artemisia annua hairy roots produce more artemisinin than diploids. Plant Cell Rep 21(8):809–813
Dhawan OP, Lavania UC (1996) Enhancing the productivity of secondary metabolites via induced polyploidy: a review. Euphytica 87(2):22280
Dixon RA (1985) Isolation and maintenance of callus and cell suspension cultures. In: Dixon RA (ed) Plant cell culture: a practical approach. IRL Press, Oxford, pp 1–20
Evans WC, Major VA (1968) The alkaloids of the genus Datura, section brugmansia. Part V. Alkaloids of D. sanguinea R. and P. and related esters of tropane-3O±, 6OI, 7OI-triol. J Chem Soc C: Organic:2775–2778
Grynkiewicz G, Gadzikowska M (2008) Tropane alkaloids as medicinally useful natural products and their synthetic derivatives as new drugs. Pharmacol Rep 60(4):439–463
Hughes EH, Shanks JV (2002) Metabolic engineering of plants for alkaloid production. Metab Eng 4(1):41–48
Ilieva M, Pavlov A (1997) Rosmarinic acid production by Lavandula vera MM cell-suspension culture. Appl Microbiol Biotechnol 47(6):683–688
Ionkova I, Witte L, Alfermann AW (1994) Spectrum of tropane alkaloids in transformed roots of Datura innoxia and Hyoscyamus x gyorffyi cultivated in vitro. Planta Med 60(4):382–384
Lavania UC (2005) Genomic and ploidy manipulation for enhanced production of phyto-pharmaceuticals. Plant Genet Resour 3(2):170–177
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479
Nussbaumer P, Kapetanidis I, Christen P (1998) Hairy roots of Datura candida x D. aurea: effect of culture medium composition on growth and alkaloid biosynthesis. Plant Cell Rep 17(5):405–409
Oksman-Caldentey KM (2007) Tropane and nicotine alkaloid biosynthesis-novel approaches towards biotechnological production of plant-derived pharmaceuticals. Curr Pharm Biotechnol 8(4):203–210
Parr AJ, Payne J, Eagles J, Chapman BT, Robins RJ, Rhodes MJC (1990) Variation in tropane alkaloid accumulation within the solanaceae and strategies for its exploitation. Phytochemistry 29(8):2545–2550
Patterson S, O’Hagan D (2002) Biosynthetic studies on the tropane alkaloid hyoscyamine in Datura stramonium; hyoscyamine is stable to in vivo oxidation and is not derived from littorine via a vicinal interchange process. Phytochemistry 61(3):323–329
Pavlov A, Kovatcheva P, Georgiev V, Koleva I, Ilieva M (2002) Biosynthesis and radical scavenging activity of betalains during the cultivation of red beet (Beta vulgaris) hairy root cultures. Z Naturforsch [C] 57(7–8):640–644
Pavlov A, Berkov S, Weber J, Bley T (2008) Hyoscyamine biosynthesis in Datura stramonium hairy root in vitro systems with different ploidy levels. Appl Biochem Biotechnol 157(2):210–225
Philipov S, Berkov S (2002) GC-MS investigation of tropane alkaloids in Datura stramonium. Z Naturforsch [C] 57(5–6):559–561
Pinol MT, Palazon J, Cusido RM, Ribo M (1999) Influence of calcium ion-concentration in the medium on tropane alkaloid accumulation in Datura stramonium hairy roots. Plant Sci 141(1):41–49
Pitta-Alvarez SI, Giulietti AM (1998) Novel biotechnological approaches to obtain scopolamine and hyoscyamine: The influence of biotic elicitors and stress agents on cultures of transformed roots of Brugmansia candida. Phytother Res 12(Suppl 1):S18–S20
Pitta-Alvarez SI, Spollansky TC, Giulietti AM (2000) The influence of different biotic and abiotic elicitors on the production and profile of tropane alkaloids in hairy root cultures of Brugmansia candida. Enzyme Microb Technol 26(2–4):252–258
Saenz-Carbonell L, Loyola-Vargas VM (1996) Datura stramonium hairy roots tropane alkaloid content as a response to changes in Gamborg’s B5 medium. Appl Biochem Biotechnol [A] 61(3):321–337
Sevon N, Oksman-Caldentey KM (2002) Agrobacterium rhizogenes-mediated transformation: Root cultures as a source of alkaloids. Planta Med 68(10):859–868
Sikuli NN, Demeyer K (1996) Influence of the ion-composition of the medium on alkaloid production by ‘hairy roots’ of Datura stramonium. Plant Cell Tissue Organ Cult 47(3):261–267
Weber J, Georgiev V, Pavlov A, Bley T (2008) Flow cytometric investigations of diploid and tetraploid plants and in vitro cultures of Datura stramonium and Hyoscyamus niger. Cytometry [A] 73(10):931–939
Witte L, Muller K, Arfmann HA (1987) Investigation of the alkaloid pattern of Datura innoxia plants by capillary gas-liquid-chromatography-mass spectrometry. Planta Med 53(2):192–197
Yamada Y, Tabata M (1997) Plant biotechnology of tropane alkaloids. Plant Biotechnol 14(1):1–10
Zabetakis I, Edwards R, O’Hagan D (1999) Elicitation of tropane alkaloid biosynthesis in transformed root cultures of Datura stramonium. Phytochemistry 50(1):53–56
Zhang L, Ding R, Chai Y, Bonfill M, Moyano E, Oksman-Caldentey KM, Xu T, Pi Y, Wang Z, Zhang H, Kai G, Liao Z, Sun X, Tang K (2004) Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures. Proc Natl Acad Sci U S A 101(17):6786–6791
Acknowledgments
This research was supported by Bulgarian Science Found under contract TK-B-1605/2006 and by EU under contract MERG-CT-2005-021344.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pavlov, A.I., Georgiev, V.G., Marchev, A.S. et al. Nutrient medium optimization for hyoscyamine production in diploid and tetraploid Datura stramonium L. hairy root cultures. World J Microbiol Biotechnol 25, 2239–2245 (2009). https://doi.org/10.1007/s11274-009-0131-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-009-0131-2