Document Type : Original Article


1 1Pharmaceutical Bioproducts Research Department, City of Scientific Research and Technology Applications, Borg-El-Arab, Egypt

2 Veg., Medicinal and Aromatic plants breeding Dept., Horticultural Research Institute (HRI), Agricultural Research Center (ARC), Egypt

3 Agriculture Botany Dept., Faculty of Agriculture (Saba Bacha), Alex. Univeristy, Egypt


Plant tissue culture techniques were used to study the effect of NaCl on tropane alkaloids accumulation and proline content of both Datura metel and Datura stramonium. Callus cultures were established for both Datura species. However, MS medium supplemented with 1mg/l of both BA and NAA was the best for callus production in D. metel, while MS supplemented with 3 mg/l Kin and 1 mg/l 2, 4-D was the optimal medium for callus production in D. stramonium. Five NaCl concentrations (25, 50, 75, 100 and 125 mM) were used to test its effect on tropane alkaloids and proline contents in the produced callus. Total tropane alkaloids were extracted from both control and salt-treated calli and analyzed using HPLC. At 125 mM NaCl scopolamine and atropine concentrations recorded 8.5 and 11.5 folds higher than control of D. metel, while they recorded 2.5 and 3.5 fold higher than control of D. stramonium after one month of culture. Peroxidase activity was determined and results showed that D. metel recorded the highest enzyme activity at 100mM NaCl concentration, while D. stramonium callus showed the highest activity at 75mM NaCl. Proline content was increased by salinity in the rate of 0.326mM proline/25mM NaCl in D. metel, and in the rate of 0.1248 mM proline/25mM NaCl in D. stramonium. The results of this investigation showed that using plant tissue culture techniques to establish callus culture from both Datura species under NaCl stress is a powerful protocol for scopolamine and atropine accumulation improvement.


Ahmed A. and E. Lffte 1970. Biosynthesis of atropine moiety of hyoscyamine from 6-Nmethylornithine. Phytochem. 9: 2345-2347.
Ahmed, A.M., M.M. Heikal and R.M. Ali 1989. Changes in amino acids and alkaloid contents in Hyoscyamus maticus and Datura stramonium in response to salinization. Phyton, 29:137-147.
Ajungla, L., P. Patil, R. Barmukh and T. Nikam 2009. Influence of biotic and abiotic elicitors on accumulation of hyoscyamine and scopolamine in root cultures of Datura metel L. Indian J. of Biotech. 8: 317-322.
Arshi, A., M.Z.Abdin and M. Iqbal 2002. Growth and metabolism of senna as affected by salt stress. Biol. Plant., 45: 295-298.
Ashraf, M. and P.J. Harris 2005. Biotic Stresses "Plant resistance through breeding and molecular approaches". Book. Pp. 47:81.
Bates, L.S., R.P. Waldeen and I.D. Teare 1973. Rapid determination of free proline for water stress studies. Plant Soil 39: 205-207.
British Pharmacopoeia, B.P. 1998. The Stationary Office Dept., BP. KE 5833 Norwich, NR 3-IBR, 1: 709-711.
Davis, R.J. 1964. Disc electrophoresis 2-method of application to human serum proteins, The Plant J. 4 (2): 215-223.
Dewick, P.M. 1997. Medicinal Natural Products: a biosynthetic approach. John Wiley and Sons, Chichester, 274-278.
Dung, N.N., E. Szoki and G. Verzar-Petri 1981. The growth dynamics of callus tissue of root and leaf origin in Datura innoxia Mill. Acta. Botanica Academiae Scientiarum Hungaricae, 27: (4), 325-333.
Ghallab, Marwa M.A. 2007. Genetical and Biochemical studies on sugar beet plants induced by tissue culture. Ph. D. Thesies. Faculty of Agric., Alex Univ., Egypt.
Huo, S.G., X.X. Gu, S.Y. Wang SY and H.X. Li 2006. Determination of scopolamine and atropine in Flos Daturae by RP-HPLC. Zhongguo Zhong Yao Za Zhi. 31(13):1065-1067.
Khlifa, Heba D.A. 2008. Genetical and Biochemical Studies on Datura metel and its Extracts. MSc Thesis. Faculty of Agriculture, University of Alexandria, Egypt.
Manchenko, G.P. 1994. Handbook of Detection of Enzyme on Electrophoretic Gel. CRG Press-Boca Raton, FL. 553 p.
Moons A., E. Prinsen, G. Bauw and M.V. Montagu 1997. Antagonistic effects of abcisis acid and jasmonates on salt stress induced transcripts in rice roots, Plant Cell, 9: 2243-2259.
Murashige, T. and F. Skoog 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant, 15: 473-497.
Nagesh, B.R. And V.R. Devaraj 2008. High temperature and salt stress response in French bean (Phaseolus vulgaris). Aust. J. Of crop Sci. 2(2): 40-48.
Nix, J. 1985. Farm Management Pocketbook, 16th ed. Wye College Farm Business Unit: Wye College Pub. 186.
Pitta-Alvarez, S., T. Spollansky and A. Giulietti 2000. The influence of different biotic and abiotic elicitors on the production and profile of tropane alkaloids in hairy root of B. candida. Enzyme Microb Technol. 26: 252-258.
Rahnama H. and H. Ebrahimzadah 2006. Antioxidant isozyme activities in potato plants (Solanium tuberosum L.) under salt stress. J. Sci. I. R. Iran. 17(3): 225-230.
Raoufa Abd El-Rahman, H.E. El-Wakil, A.E. Abou Gabal and H.D. Khlifa 2008. Production of Scopolamine and Hyoscyamine in Calli and Regenerate Cultures of Datura metel (L.). J. of App. Sci. Res.4(12): 1858-1866.
Reddy M.P. and A.B. Vora 1985. Effect of salinity on protein metabolism in bajora (Pennisetum typhoides) leaves. Indian J. Plant Physiol. 28: 190-198.
Silveira J.A.G., R.A. Viegas, I.M.A. Rocha, A.C.O. Monteiro-Moreira, R.M. Moreira and J.T.A Oliveira 2002. Proline accumulation and glutamine synthetase are increased by salt-induced proteolysis in cashew leaves. J. Plant Physiol.
Smith, R.H. 2000. Callus induction in plant tissue culture techniques and experiments. 2nd ed. Acad. Press, San Diego, California pp. 98-103.
Stewart, C.R. and A.D. Hanson 1980. Proline accumulation as a metabolic response to water stress. In: TURNER N. C. & KRAMER P. J., Adaptation of plants to water and high temperature stress, New York. pp. 173-189.
Stewart, C.R. and J.A. Lee 1974. The role of proline accumulation in haplophytes. Planta. 120: 279-289.
Warrier, P.K., V.P.K Nambiar and C. Ramankutty 1994. Indian medicinal plants: a compaendium of 500 species, Orient Longman, Hyderabad. 2: 164-172.
Weber, D., B. Clare and M.Stahmanm 1976. Ezymic changes associated with induced and natural resistance of sweet potato to Ceratocystis fimoriata. Phyto. 57 (4): 241-424.
Zhang, L., R. Ding, Y. Chai, M. Bonfill, M. Piaol, T. Xu, Y. Pi, Z. Wang, H. Zhang, G.Y. Kai, Z.H. Liao, X.F. Sun and K. Tang 2004. Engineering tropane biosynthetic pathway in Hyoscyamus niger hairy root cultures. Proc. Natl. Acad. Sci. USA 101: 6786-6791.