In collaboration with Scientific Association of Iranian Medicinal Plants

Document Type : Research Paper

Authors

Department of Plant Physiology and genetic, Applied Science Institute, Jahad-e-Daneshgahi (ACECR), Shahid Beheshti University, Tehran, Iran

Abstract

Tropane alkaloids such as atropine and scopolamine have wide application in the treatment of diseases such as asthma and antispasmodic due to anticholinergic agents. In the present study, the effects of salicylic acid (SA) on the production of  two alkaloids, atropine and scopolamine ,were studied in hairy root and in vitro grown root cultures of Hyoscyamus niger L. The roots were cultured in liquid B5 medium containing different concentrations of SA (0, 0. 1, 1, 2 and 4 mM) in various exposure times (24, 96 and 168 hours). Eventually, root growth index, and atropine and scopolamine content were assayed after 30 days. In in vitro grown roots, treatment with 1mM SA resulted in the highest production of atropine after 168 hours, while the highest amount of scopolamine (649.53 µg/g D. W) was obtained in 2mM SA (after 96 hours), showing more than 13-fold increase compared to the control. In hairy root cultures, the most significant contents of atropine were observed in the medium containing 2mM SA after 96 h. Moreover, the highest content of scopolamine was achieved in medium treated with 0.1 mM SA after 96 hours. In general, atropine content in hairy roots was considerably higher than that of in vitro grown roots. In contrast, scopolamine content in in vitro grown roots was significantly more than that of hairy roots. Moreover, the rate of root growth declined as a result of increasing of elicitor concentration in the medium. 

Keywords

- Ahmadian Chashmi, N., Sharifi, M., Karimi, F. and Rahnama, H., 2010. Comparative study of tropane alkaloids production in hairy roots and plantlet cultures of Atropa belladonna L. by salicylic acid treatments. Iranian Journal of Plant Biology, 1(3): 63-76.
- Ajungla, L., Patil, P.P., Barmukh, R.B. and Nikam, T.D., 2009. Influence of biotic and abiotic elicitors on accumulation of Hyoscyamine and scopolamine in root cultures of Datura metel L. Indian Journal of Biotechnology, 8: 317-322.
- Akramian, M., Fakhr Tabatabaei, S.M. and Mirmasoumi, M., 2008. Virulence of Different Strains of Agrobacterium rhizogenes on Genetic Transformation of Four Hyoscyamus Species. American-Eurasian Journal of Agricultural & Environmental Sciences, 3: 759-763.
- Biondi, S., Fornalé, S., Oksman-Caldentey, K.M., Eeva, M., Agostani, S. and Bagni, N., 2000. Jasmonates induce over-accumulation of methylputrescine and conjugated polyamines in Hyoscyamus muticus L. root cultures. Plant Cell Reports, 19: 691-697.
- Bruce, N., 2008. Alkaloids: 332-350. In: Rehm, H.J., Reed, G. and Bruce N.C., (Eds.). Biotechnology Set. Wiley, Cambridge, UK.
- Bulgakov, V.P., Tchernoded, G.K., Mischenko, N.P. and Khodakovskaya, M.V., 2002. Effect of salicylic acid, methyl jasmonate, ethephon and cantharidin on anthraquinone production by Rubia cordifolia callus cultures transformed with the rolB and rolC genes. Journal of Biotechnology, 97: 213-221.
- Cai, D., Kleine, M., Kifle, S., Horloff, H.J., Sandal, N.N., Marcker, K.A., Lankhorst, R.M.K., Salentijn, E.M.J., Lange, W., Stiekema, W.J., Wyss, V., Grundler, F.M.W. and Jung, C., 1997. Positional cloning of a gene for nematode resistance in sugarbeet. Science, 275: 832-834.
- Ebrahimzadeh, H., Teimoori, A. and Lohrasbi, T., 2003. Hyoscyamin 6-β-hydroxylase gene isolation from in vitro cultured roots of Hyoscyamus niger L. and Hyoscyamus tenuicaulis. Daru, 11: 34-37.
- Gamborg, O.L., Miller, R.A. and Ojima, K., 1968. Nutrient requirements of suspension cultures of soybean root cells. Experimental Cell Research, 50: 151-158.
- Ghanati, F., Bakhtiarian, S. and Abdolmaleki, P., 2010. Effect of methyl jasmonate on second metabolites in (Calendula officinalis L.). Modares Biological Sciences and Technology, 1(1): 21-31.
- Grynkiewicz, G. and Gadzikowska, M., 2008. Tropane alkaloids as edicinally useful natural products and their synthetic derivatives as new drugs. Pharmacological Reports, 60: 439-463.
- Grynkiewicz, M. and Gadzikowska, G., 2002. Tropane alkaloids in pharmaceutical and phytochemical analysis. Acta Polonine pharmaceuica, 59(2): 149-160.
- Kai, G., Yang, Sh., Zhang, Y., Luo, X., Fu, X., Zhang, A. and Xiao, J., 2012. Effects of different elicitors on yield of tropane alkaloids in hairy roots of Anisodus acutangulus. Molecular Biology Reports, 39: 1721-1729.
- Kamada, H., Okamura, N., Satake, M., Harada, H. and Shimomura, K., 1986. Alkaloid production by hairy root cultures in Atropa belladonna. Plant Cell Reports, 5: 239-242.
- Kang, S.M., Jung, H.Y. and Kang Y.M., 2004. Effect of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of Scopolia parviflora. Plant Science, 166(3): 745-751.
- Khatamsaz, M., 1998. Flora of Iran (Solanaceae No. 24). Research Institute of Forests and Rangeland Press, Tehran, 116p.
- Mizukami, H., Tabira, Y. and Ellis, B.E., 1993. Methyl jasmonate-induced rosmarinic acid biosynthesis in Lithospermum erythrorhizon cell suspension cultures. Plant Cell Reports, 12: 706-709.
- Murashige, T. and Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497.
- Namdeo, A.G., 2007. Plant cell elicitation for production of secondary metabolites. Pharmacognosy Reviews, 1(1): 69-79.
- Palazón, J., Navarro-Ocaña, A., Hernandez-Vazquez, L. and Mirjalili, M.H., 2008. Application of metabolic engineering to the production of scopolamine. Molecules,13(8): 1722-1742.
- Parsa, M., Garoosi, G.A. and Haddad, R., 2011a. Effect of jasmonate and methyl jasmonate elicitors on quantity and quality of total RNA extraction in Hyoscyamus niger. The 2nd of National Conference of Plant Physiology, Yazd, Iran, 28-29 April.
- Parsa, M., Zeinali, A. and Yousefzadi, M., 2011b. Induction of hairy roots with two strains of Agrobacterium rhizogenes (A4 and LBA9402) in Hyoscyamus niger. The 2nd of National Conference of Plant Physiology, Yazd, Iran, 28-29 April.
- Roos, R.W. and Lau-Cam, C., 1986. General reversed-phase HPLC method for the separation of drugs using triethyamine as a competing base. Journal of Chromatography, 370: 403-418.
- Wang, C., Wu, J. and Mei, X., 2001. Enhanced taxol production and release in Taxus chinensis cell suspension cultures with selected organic solvents and sucrose feeding. Biotechnology Progress, 17(1): 89-94.
- Wielanek, M. and Urbanek, H., 2006. Enhanced glucotropaeolin production in hairy root cultures of Tropaeolum majus L. by combining elicitation and precursor feeding. Plant Cell of Tissue Organism, 86: 177-186.
- Yan, Q., Shi, M., Ng, J. and Yong, J., 2006. Elicitor-induced Rosmarinic Acid Accumulation and Secondary Metabolism Enzyme Activities in Salvia miltiorrhiza Hairy Roots. Plant Science, 170: 853-858.
- Zabetakis, I., Edwardsb, R. and O'Hagana, D., 1999. Elicitation of tropane alkaloid biosynthesis in transformed root cultures of Datura stramonium. Phytochemistry, 50: 53-56
- Zayed, R. and Winka, M.Z., 2004. Induction of tropane alkaloid formation in ransformed root cultures of Brugmansia suaveolens (Solanaceae). Zeitschrift für Naturforschung C. A Journal of Biosciences, 59: 863-867.
- Zolala, J., Farsi, M., Gordan, H. R. and Mahmoodnia, M., 2007. Producing a high scopolamine hairy root clone in Hyoscyamus muticus through transformation by Agrobacterium rhizogenes. Journal of Agricultural Sciences of Technology, 9: 327-339.