Effect of proline and salicylic acid on morphological and phytochemical characteristics of Stevia rebaudiana (Bert.)

Document Type : Full Paper


1 Former M.Sc. Student, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

2 Assistant Professor, Faculty of Agriculture, University of Zanjan, Zanjan, Iran


To study the effect of foliar application of proline and salicylic acid on growth, morphological and phytochemical traits of Stevia, an experiment based on completely randomized block design in a factorial arrangement was conducted with three replications in the University of Zanjan. Foliar applications of proline in three levels (0, 1, 2 mM) as well as salicylic acid (0, 0.3, 0.6 mM) were applied after establishment of transplants in the field three times with 10 days intervals. Factors including leaf area, leaf fresh and dry weight, total fresh and dry weight, chlorophyll a, b and total chlorophyll, carotenoid, total phenol, total flavonoid and antioxidant activity were studied. Results showed that salicylic acid and proline applications significantly influenced above mentioned traits. The highest average leaf area/ plant (5949.39 cm2), leaf fresh (787.17 g/m2) and dry leaf weight (228.5 g/m2), total fresh (1339.23 g/m2) and total dry weight (327.52 g/m2) were recorded in 1 mM proline in combination with 0.3 mM salicylic acid. Solely applications of proline at 2 mM and salicylic acid at 0.6 mM enhanced chlorophyll a, b, carotenoid and total flavonoid contents. The highest average total chlorophyll (5.23 mg/g fw) observed in 2 mM proline in combination with 0.3 mM salicylic acid and highest phenol (19.94 mg/g dw) recorded in 1 mM proline in combination with 0.6 mM salicylic acid. Collectively, results showed that application of proline and salicylic acid significantly improved vegetative growth and production of active substances in Stevia rebaudiana.


Main Subjects

  1. Ashton, J. D. & Deshpal, S. V. (1993). Proline biosynthesis and osmoregulation in plants. The Plant Journal, 4(2), 215-223.
  2. Ashraf, M. Y., Azmi, A. R., Khan, A. H. & Ala, S. A. (1994). Effect of water on total phenols, peroxidase activity and chlorophyll content in wheat. Acta Phsiologiae Plantarum, 16, 185-191.
  3. Ali, M. B., Hahn, E. J. & Paek, K. Y. (2007). Methyl jasmonat and salicylic acid induced oxidative stress and accumulation of phenolic in Panax ginseng Bioreactor root suspension culture. Journal Molecules, 12, 607-621.
  4. Azarpur, E., Motamed, K. M. & Bozorgi, H. R. (2013). Stevia agronomy and extension (Botany, cultivation, maintenance, harvesting, chemistry, industry and processing). (1st ed). Azad University of Lahijan. (in Farsi)
  5. Abd Elhamid, E. M., Sadak, M. S. & Tawfik, M. M. (2016). Physiological response of Fenugreek plant to the application of proline under different water regimes. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 7(3), 580-594.
  6. Barker, D. J., Sullivan, C. Y. & Moser, L. E. (1993). Water deficit effects on osmotic potential, cell wall elasticity and proline in five forage grasses. Agronomy Journal, 85, 270-275.
  7. Brand-Williams, W., Cuvelier, M. E. & Berset, C. (1995). Use of a free radical method to evaluate antioxidant activity, Lebensmittel Wissenschaft und Technologie, 28, 25-30.
  8. Boukraâ, D., Belabid, L., Benabdelli, K. & Bennabi, F. (2015). Implication of salicylic acid in chickpea growth to salt resistance. Advances in Environmental Biology, 9(27), 270-277.
  9. Dragovi-Uzelac, V., Savi, Z., Brala, A., Levaj, B., BursaKovaevi, D. & Bisko, A. (2010). Evaluation of phenolic content and antioxidant capacity of blueberry cultivars (Vaccinium corymbosum L.) grown in the northwest Croatia. Food Technology and Biotechnology, 48(2), 214-221.
  10. El-Tayeb, M. A. (2005). Response of Barley grains to the interactive effect of salinity and salicylic acid. Plant Growth Regulation, 45(6), 215-255.
  11. Gamal El-Din, K. M. & Abd El-Wahed, M. S. A. (2005). Effect of some amino acids on growth and essential oil content of chamomile plant. International Journal of Agricultural and Biological Engineering, 7, 376-380.
  12. Gharib, F. A. L. (2006). Effect of salicylic acid on the growth, metabolic activities and oil content of basil and majoram. International Journal of Agriculture and Biology, 4, 485-492.
  13. Ghasemi Pirbaloti, A., Mousavi Haris, S. A., Tyrksh, F. & Hamedi, B. (2012). Effect of Jasmonic acid and Salicylic acid on phenolic compounds and flavonoids of flower extract in Calendula officinalis. Journal of Herbal Drugs, 3(3), 175-180. (in Farsi)
  14. Ghasemi, S. & Ghasemi-Golezani, K. (2015). Assessment of leaf number and chlorophyll content of Carum copticom in response to salicylic acid and abscisic acid. International Conference on Sustainable Development, Strategies and Challenges with a Focus on Agriculture, Natural Resources, Environment and Tourism. Tabriz, Iran. (in Farsi)
  15. Hirose, T., Ackerly, D. D., Traw, M. B., Ramseier, D. & Bazzaz, F. A. (1997). CO2 elevation, canopy photosynthesis and optimal leaf area index. Ecology, 78, 2339-50.
  16. Khan, W., Prithiviraj, B. & Smith, D. (2003). Photosynthetic responses of corn and soybean to foliar application of salicylates. Plant Physiology, 160, 485-92.
  17. Khan, N. A., Syeed, S., Masood, A., Nazar, R. & Iqbal, N. (2010). Application of salicylic acid increases contents of nutrients and antioxidative metabolism in mungbean and alleviates adverse effects of salinity stress. International Journal of Plant Biology, 1(5), 1-8.
  18. Khodary, S. F. A. (2004). Effect of salicylic acid on the growth, photosynthesis and carbohydrate metabolism in salt stressed maize plants. International Journal of Agriculture and Biology, 6, 5-8.
  19. Kaydan, D., Yagmur, M. & Okut, N. (2007). Effects of salicylic acid on the growth and some physiological characters in salt stressed wheat (Triticum aestivum L.). Tarim Bilimleri Dergisi, 13(2), 114-119.
  20. Khandaker, L., Masum Akond, A. S. M .G. & Oba, S. (2011). Foliar application of salicylic acid improved the growth, yield and leaf’s bioactive compounds in red amaranth (amaranthus tricolor L.).Unauthenticated, 74, 77-86.
  21. Kahlaoui, B., Hachicha, M., Teixeira, J., Misle, E., Fidalgo, F. & Hanchi, B. (2013). Response of two tomato cultivars to field-applied proline and salt stress. Journal of Stress Physiology and Biochemistry, 9, 357-365.
  22. Mahtabi, G., Fazeli Fard, M. H. & Majnooni, A. (2013). Climate classification of Zanjan state. (2013). First National Conference on Climatology of Iran. Kerman Graduate University of Advanced Technology. (in Farsi)
  23. Metwally, A., Finkemeier, I., Georgi, M. & Dietz, k. J. (2003). Salicylic acid alleviates the cadmium toxicity in barley seedlings. Plant Physiology, 132(1), 273 -281.
  24. Meda, A., Lamien, C. E., Romito, M., Millogo, J. & Nacoulma, O. G. (2005). Determination of the total phenolic, flavonoid and praline contents in Burkina Fasan honey, as well as their scavenging activity. Food Chemistry, 91, 571-577.
  25. El Far, M. M. & Taie, H. A. (2009). Antioxidant activities, total anthocyanins, phenolic and flavonoids contents of some sweet potato genotypes under Stress of different concentrations of sucrose and sorbitol. Australian Journal of Basic and Applied Sciences, 3(4), 3609-3616.
  26. Mueller, M. J., Brodschelm, W., Spannagl, E. & Zenk, M. H. (1993). Signaling in the elicitation process is mediated through the octadecanoid pathway leading to jasmonic acid. In: Proceedings of the National Academy of Sciences of the United States of America, 90, 7490-7494.
  27. Messedi, D., Farhani, F., Ben-Hamid, K., Trablsi, N., Ksouri, R., Ather, H. U. R. & Abdelly, C. (2016). Highlighting the mechanisms by which proline can confer tolerance to salt stress in Cakile maritima. Pakistan Journal of Botany, 48(2), 417-427.
  28. Pacheco, A. C., Silva Cabral, C. D., Silva Fermino, E. S. D. & Aleman, C. C. (2013). Salicylic acid-induced changes to growth, flowering and flavonoids production in marigold plants. Journal of Medicinal Plant Research, 7(42), 3158-3163.
  29. Pandey, M. & Chikara, S. K. (2014). In vitro Regeneration and effect of abiotic stress on physiological and biochemical content of Stevia Rebaudiana (Bertoni). Journal of Plant Science Research, 1(3), 113.‏
  30. Qinghua, S. H. & Zhujun, Z. (2008). Effect of exogenous salicylic acid on manganese toxicity, element contents and antioxidative system in cucumber. Environmental and Experimental Botany, 63, 317-326.
  31. Raskin, I. (1992). Role of salicylic acid in plants. Annual Reviews of Plant Physiology and Plant Molecular Biology, 43,439-463.
  32. Ramesh, K., Singh, V. & Megeji, N. W. (2006). Cultivation of Stevia rebaudiana (BERT), a comprehensive review.Advances in Agronomy, 89, 137-177.
  33. Ryahy, N., Farahbakhsh, H. & Pasandy Pour, A. (2011). Foliar application of proline, glycine betaine, salicylic acid, ascorbic acid mitigate the drought effects on sorghum, 11th Congress on Irrigation and evaporation reduction. Shahid Bahonar University of Kerman, Iran. (in Farsi)
  34. Rashad EL-Sherbeny, M. & Teixeira da Silva, J. A. (2013). Foliar treatment with proline and tyrosine effect the growth and yield of beetroot and some pigments in beetroot leaves. Journal of Horticultural Research, 21(2), 95-99.
  35. Ruiz Ruiz, J. C., Moguel Ordonez, Y. B., Basto, A. M. & Segura Campos, M. R. (2015). Antioxidant capacity of leaf extracts from two Stevia rebaudiana Bertoni varieties adapted to cultivation in Mexico. Nutricion Hospitalaria, 31(3), 1163-1170.
  36. Shukla, S., Mehta, A. & Bajpai, V. K. (2009). In vitro antioxidant activity and total phenolic content of ethanolic leaf extract of Stevia rebaudiana Bert. Food and Chemical Toxicology, 47, 2338-2343.
  37. Shbani, L. & Ehsanpour, A. (2009). Induced antioxidant enzymes, phenolic compounds and flavonoids under in vitro cultivation of Glycrrihiza glabra. Biological Science Promotion, 22(4), 691-703. (in Farsi)
  38. Singh, S. D. & Rao, G. P. (2005). Stevia: The herbal sugar of the 21st century. Sugar Technology, 7, 17-24.
  39. Szabados, L. & Savoure, A. (2009). Proline: a multifunctional amino acid. Trends in Plant Science, 15(2), 82-97.
  40. Sadeghipour, O. & Aghaei, P. (2012). Impact of exogenous salicylic acid application on some traits of common bean (Phaseolus vulgaris L.) under water stress conditions. International Journal of Agriculture and Crop Sciences, 4(11), 685-690.
  41. Tadhani, M. B., Patel, V. H. & Subhash, R. (2007). In vitro antioxidant activities of Stevia rebaudiana leaves and callus. Journal of Food Composition and Analysis, 20, 323-329.
  42. Yadav, A. K., Singh, S., Dhyani, D. & Ahuja, P. S. (2011). A review on the improvement of Stevia. Stevia rebaudiana (Bertoni). Canadian Journal Plant Science, 91, 1-27.
  43. Yousry, M. M., El-Mesirry, D. S. & Shama, M. A. (2015). Effect of Proline on Resistance of Potato Crop (Solanum tuberosum L.) for the Negative Effects of Water Irrigation Salinity. Current Science International, 4(1), 172-177.
  44. Zawoznik, M. S., Gropp, M. D., Tomaro, M. & Benavides, M. P. (2007). Endogenous salicylic acid potentiates cadmium- induced oxidative stress in Arabidopsis thaliana. Journal Plant Science, 173, 190-197.
  45. Zhang, C. S., Lu, Q. & Verma, D. P. S. (1997). Characterization of Δ-1-pyrroline-5-carboxylatesynthetase gene promoter in transgenic Arabidopsis thaliana subjected to water stress. Plant Science, 129, 81-89.