Sugar Tech

, Volume 21, Issue 6, pp 1031–1038 | Cite as

Effect of Methyl Jasmonate and Salicylic Acid on In Vitro Growth, Stevioside Production, and Oxidative Defense System in Stevia rebaudiana

  • Sajjad MoharramnejadEmail author
  • Amirali Taheri Azam
  • Jaber Panahandeh
  • Zahra Dehghanian
  • Muhammad Ashraf
Research Article


To examine whether secondary metabolites in micro-propagated Stevia rebaudiana plants are enhanced, explants were cultured on the woody plant medium (WPM) supplemented separately with methyl jasmonate (MeJA) and salicylic acid (SA). Maximal shoot growth, root regeneration, and fresh and dry biomass accumulation were obtained on WPM containing 50 μM SA. Electrophoretic analysis of superoxide dismutase (SOD), peroxidase (POX), and catalase (CAT) in S. rebaudiana showed that three, two, and one isoforms were observed for SOD, POX, and CAT, respectively. The WPM containing MeJA and SA increased SOD and POX activities, especially those of SOD2 and POX1 isoforms, total phenolics and total antioxidant activity (DPPH; 1-1-diphenyl 2-picryl hydrazyl), and decreased CAT activity. The highest activities/levels of enzymatic and non-enzymatic antioxidants were observed in the WPM containing 100 μM SA. The high-performance liquid chromatography (HPLC) analyses showed that 50 μM MeJA and 100 μM SA supplementation enhanced stevioside production. The stevioside yield increased from 4.1 mg/g dry weight to 18.7 and 19.4 mg/g dry weight in in vitro plantlets exposed to 50 μM MeJA and 100 μM SA, respectively. The results showed that SA could regulate oxidative defense system and secondary metabolite production in micro-propagated S. rebaudiana.


Antioxidant activity Culture Electrophoretic Elicitor Phenolic 


Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ali, M.B., K.W. Yu, E.J. Hahn, and K.Y. Paek. 2006. Methyl jasmonate and salicylic acid elicitation induces ginsenosides accumulation, enzymatic and non-enzymatic antioxidant in suspension culture Panax ginseng roots in bioreactors. Plant Cell Reports 25 (6): 613–620. Scholar
  2. Ashraf, M. 2010. Inducing drought tolerance in plants: Recent advances. Biotechnology Advances 28 (1): 169–183. Scholar
  3. Bayraktar, M., E. Naziri, I.H. Akgun, F. Karabey, E. Ilhan, B. Akyol, E. Bedir, and A. Gurel. 2016. Elicitor induced stevioside production, in vitro shoot growth, and biomass accumulation in micropropagated Stevia rebaudiana. Plant Cell, Tissue and Organ Culture 127 (2): 289–300. Scholar
  4. Chong, T., M. Abdullah, N. Fadzillah, O. Lai, and N. Lajis. 2005. Jasmonic acid elicitation of anthraquinones with some associated enzymic and non-enzymic antioxidant responses in Morinda elliptica. Enzyme and Microbial Technology 36 (4): 469–477. Scholar
  5. Chung, I.-M., M. Thiruvengadam, K. Rekha, and G. Rajakumar. 2016. Elicitation enhanced the production of phenolic compounds and biological activities in hairy root cultures of bitter melon (Momordica charantia L.). Brazilian Archives of Biology and Technology 59: 1–10. Scholar
  6. Kim, H.-J., F. Chen, X. Wang, and J.-H. Choi. 2006. Effect of methyl jasmonate on phenolics, isothiocyanate, and metabolic enzymes in radish sprout (Raphanus sativus L.). Journal of Agricultural and Food Chemistry 54 (19): 7263–7269. Scholar
  7. Lloyd, G., and B. McCown. 1980. Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Combined Proceedings, International Plant Propagators’ Society 30: 421–427.Google Scholar
  8. Lucho, S.R., M.N. do Amaral, A. López-Orenes, A.M. Kleinowski, L. do Amarante, M.Á. Ferrer, A.A. Calderón, and E.J.B. Braga. 2018a. Plant growth regulators as potential elicitors to increase the contents of phenolic compounds and antioxidant capacity in stevia plants. Sugar Tech. Scholar
  9. Lucho, S.R., M.N. do Amaral, C. Milech, M.Á. Ferrer, A.A. Calderón, V.J. Bianchi, and E.J.B. Braga. 2018b. Elicitor-induced transcriptional changes of genes of the steviol glycoside biosynthesis pathway in Stevia rebaudiana Bertoni. Journal of Plant Growth Regulation 1: 1–15. Scholar
  10. Mathur, S., and G.S. Shekhawat. 2013. Establishment and characterization of Stevia rebaudiana (Bertoni) cell suspension culture: an in vitro approach for production of stevioside. Acta Physiologiae Plantarum 35 (3): 931–939. Scholar
  11. Mittler, R. 2002. Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science 7 (9): 405–410. Scholar
  12. Moharramnejad, S., O. Sofalian, M. Valizadeh, A. Asgari, and M. Shiri. 2015. Proline, glycine betaine, total phenolics and pigment contents in response to osmotic stress in maize seedlings. Journal of Bioscience and Biotechnology 4 (3): 313–319.Google Scholar
  13. Moharramnejad, S., O. Sofalian, M. Valizadeh, A. Asgari, and M. Shiri. 2016. Response of antioxidant defense system to osmotic stress in maize seedlings. Fresenius Environmental Bulletin 25 (3): 805–811.Google Scholar
  14. Muffler, K., D. Leipold, M.-C. Scheller, C. Haas, J. Steingroewer, T. Bley, H.E. Neuhaus, M.A. Mirata, J. Schrader, and R. Ulber. 2011. Biotransformation of triterpenes. Process Biochemistry 46 (1): 1–15. Scholar
  15. Olson, P., and J. Varner. 1993. Hydrogen peroxide and lignification. The Plant Journal 4 (5): 887–892. Scholar
  16. Pérez-Alonso, N., A. Capote, A. Gerth, and E. Jiménez. 2012. Increased cardenolides production by elicitation of Digitalis lanata shoots cultured in temporary immersion systems. Plant Cell, Tissue and Organ Culture 110 (1): 153–162. Scholar
  17. Raman, V., and S. Ravi. 2011. Effect of salicylic acid and methyl jasmonate on antioxidant systems of Haematococcus pluvialis. Acta Physiologiae Plantarum 33 (3): 1043–1049. Scholar
  18. Razak, U.N.A.A., C.B. Ong, T.S. Yu, and L.K. Lau. 2014. In vitro micropropagation of Stevia rebaudiana Bertoni in Malaysia. Brazilian Archives of Biology and Technology 57 (1): 23–28. Scholar
  19. Ruiz Ruiz, J.C., Y.B. Moguel Ordoñez, Á. Matus Basto, and M.R. Segura Campos. 2015. Antioxidant capacity of leaf extracts from two Stevia rebaudiana Bertoni varieties adapted to cultivation in Mexico. Nutricion Hospitalaria 31 (3): 1163–1170. Scholar
  20. Sharma, M., A. Ahuja, R. Gupta, and S. Mallubhotla. 2015. Enhanced bacoside production in shoot cultures of Bacopa monnieri under the influence of abiotic elicitors. Natural Product Research 29 (8): 745–749. Scholar
  21. Shukla, S., A. Mehta, V.K. Bajpai, and S. Shukla. 2009. In vitro antioxidant activity and total phenolic content of ethanolic leaf extract of Stevia rebaudiana Bert. Food and Chemical Toxicology 47 (9): 2338–2343. Scholar
  22. Sivanandhan, G., G.K. Dev, M. Jeyaraj, M. Rajesh, A. Arjunan, M. Muthuselvam, M. Manickavasagam, N. Selvaraj, and A. Ganapathi. 2013. Increased production of withanolide A, withanone, and withaferin A in hairy root cultures of Withania somnifera (L.) Dunal elicited with methyl jasmonate and salicylic acid. Plant Cell, Tissue and Organ Culture 114 (1): 121–129. Scholar
  23. Soleymani, F., and H. Taheri. 2015. Effects of gibberellic acid, methyl jasmonate and chitosan on antioxidant enzyme activity in peppermint (Mentha piperita L.). Journal of Advances in Chemical Engineering and Biological Sciences 2 (2): 135–138. Scholar
  24. Soltis, D.E., P.S. Soltis, and T.R. Dudley. 2012. Isozymes in plant biology. Dordrecht: Springer.Google Scholar
  25. Tadhani, M., V. Patel, and R. Subhash. 2007. In vitro antioxidant activities of Stevia rebaudiana leaves and callus. Journal of Food Composition and Analysis 20 (3–4): 323–329. Scholar
  26. Uddin, M.S., M.S.H. Chowdhury, M.M.M.H. Khan, M.B. Uddin, R. Ahmed, and M. Baten. 2006. In vitro propagation of Stevia rebaudiana Bert in Bangladesh. African Journal of Biotechnology 5 (13): 1238–1240.Google Scholar
  27. Valizadeh, M., S. Moharamnejad, M. Ahmadi, and H. Mohammadzadeh Jalaly. 2013. Changes in activity profile of some antioxidant enzymes in alfalfa half-sib families under salt stress. Journal of Agricultural Science and Technology 15 (4): 801–809.Google Scholar
  28. Xu, S., W. Zhou, S. Pottinger, and I.T. Baldwin. 2015. Herbivore associated elicitor-induced defences are highly specific among closely related Nicotiana species. BMC Plant Biology 15 (2): 1–13. Scholar
  29. Zayova, E., I. Stancheva, M. Geneva, M. Petrova, and L. Dimitrova. 2013. Antioxidant activity of in vitro propagated Stevia rebaudiana Bertoni plants of different origins. Turkish Journal of Biology 37 (1): 106–113. Scholar
  30. Zhao, J., L.C. Davis, and R. Verpoorte. 2005. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnology Advances 23 (4): 283–333. Scholar

Copyright information

© Society for Sugar Research & Promotion 2019

Authors and Affiliations

  1. 1.Crop and Horticultural Science Research Department, Ardabil Agricultural and Natural Resources Research and Education CenterAREEOMoghanIran
  2. 2.Department of Plant Breeding and Biotechnology, Faculty of AgricultureUniversity of TabrizTabrizIran
  3. 3.Department of Horticultural Science, Faculty of AgricultureUniversity of TabrizTabrizIran
  4. 4.Institute of Molecular Biology and TechnologyThe University of LahoreLahorePakistan

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