Document Type : Research paper


Department of Horticulture Sciences, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran


To study effects of salicylic acid (SA) treatments (0, 0.5, 1 and 2 mM) in two forms of seed priming (for 24 h) and spraying on seedling at 4-6 leaf stage an experiment was conducted on periwinkle (Catharanthus roseus Don.) with and without exposure to a mild cold stress of 8 °C (for 32 h). Seeds were sown in trays containing peat-based substrate (with 20% sandy loam field soil). Transplants of both groups were treated with mild cold. Seedlings were grown in greenhouse with an average day/night temperature of 25/20± 2°C until their blooming. Seed emergence rate and percentage, activities of catalase (CAT), guaiacol peroxidase (GPX), polyphenol oxidase (PPO), CAT gene expression, height and number of nodes per plant, and days to flowering were evaluated. Results showed that mild cold stress together with salicylic acid at 0.5 mM (as either seed priming or spray on seedling) was the best treatment to accelerate the flowering, and improve growth parameters and antioxidant enzymatic activities. Thus, mild cold stress enhanced the positive effects of SA treatments on cold acclimation of periwinkle (particularly when associated with spraying, which is a simple administration method). Accordingly, it might be recommended for its seedling production and cultivation in temperate climates to prevent late spring frost damages.


  1. Aebi H. 1984. Catalase in vitro. In: Methods in enzymology. Florida, Academic Press 114-121.
  2. Agarwal S, Sairam R.K, Srivastava G.C, Meena R.C. 2005. Changes in antioxidant enzymes activity and oxidative stress by abscisic acid and salicylic acid in wheat genotypes. Biologia Plantarum 49, 541–550.
  3. Ananieva E.A, Alexieva V.S, Popova L.P. 2002. Treatment with salicylic acid decreases the effects of paraquat on photosynthesis. Journal of Plant Physiology 159, 685-693.
  4. Apostolova P, Yaneva I. 2006. Antioxidative defense in winter wheat plants during early cold acclimation. General and Applied Plant Physiology, Special issue 101-108.
  5. Basler D, Körner C. 2012. Photoperiod sensitivity of bud burst in 14 temperate forest tree species. Agricultural and Forest Meteorology165, 73–81.
  6. Belkhadi A, Hediji H, Abbes Z, Nouairi I, Barhoumi Z, Zarrouk M, Chaibi W, Djebali W. 2010. Effects of exogenous salicylic acid pre-treatment on cadmium toxicity and leaf lipid content in Linum usitatissimum L. Ecotoxicology and Environmental Safety 73, 1004-1011.
  7. Borsani O, Valpuesta V, Botella M.A. 2001. Evidence for a role of salicylic acid in the oxidative damage generated by NaCl and osmotic stress in arabidopsis seedlings. Plant Physiology 126, 1024–1030.
  8. Brevendan R.E, Egli D.B, Leggett J.E. 1978. Influence of N nutrition on flower and pod abortion and yield of soybeans. Agronomy Journal 70, 81-84.
  9. Cervilla L.M, Rosales M.A, Rubio-Wilhelmi M.M, Sanchez-Rodriguez E, Blasco B, Rios J.J. 2009. Involvement of lignification and membrane permeability in the tomato root response to boron toxicity. Plant Science 176, 545–552.
  10. Chance B, Maehly A.C. 1955. Assay of catalase and peroxidases. Methods in Enzymology 2, 764-775.
  11. Chang C.J, Koa C.H. 1988. H2O2 metabolism during senescence of rice leaves changes in enzyme activities in light and darkness. Plant Growth Regulation 25, 11–15.
  12. Chuine I, Morin X, Bugmann H. 2010. Warming, photoperiods, and tree phenology. Science 329, 277–278.
  13. Dat J.F, Lopez-Delgado H, Foye C.H, Scott I.M. 2000. Effect of salicylic acid on oxidative stress and thermo tolerance in tobacco. Journal of Plant Physiology 156, 659-665.
  14. DeKock P.C, Grabowsky F.B, Innes, A.M. 1974. The effect of salicylic acid on the growth of Lemna gibba. Annals of Botany 38, 908-903.
  15. De Villiers A.J, Van Rooyrn M.W, Theron G.K, Van Deventer H.A. 1994. Germination of three namaqualand pioneer species, as Influenced by Salinity, temperature and light. Seed Science and Technology 22, 427-433.
  16. Dole J.M, Wilkins H.F. 1999. Floriculture: Principles and Species. Upper saddle river, N.J. Prentice Hall.
  17. Drazic G, Mihailovic N, Lojic M. 2006. Cadmium accumulation in Medicago sativa seedlings treated with salicylic acid. Biologia Plantarum 50, 239-244.
  18. El-Domyati F.M, Abdel Azeiz A.Z,  Atef A, Elhalwagi A, Sabir J.S.M, Ramadan A.M, Gadalla N.O, Edris S, Baeshen M.N, Hajrah N.H, Al-Kordy M.A, Al-Hajar A.S.M,  Akoh C.C, Bahieldin A. 2014. Functional genomics analysis for overproduction of key terpenoid indole alkaloids in mature leaf of Catharanthus roseus. Life Science Journal 11(10), 1213-1220.
  19. Fariduddin Q, Hayat S, Ahmad A. 2003. Salicylic Acid Influences net photosynthetic rate, carboxylation efficiency, nitrate reductase activity, and seed yield in Brassica juncea. Photosynthetica 41, 281-284.
  20. Farooq M, Aziz T, Basra S.M.A, Cheema M.A, Rehman H. 2008. Chilling tolerance in hybrid maize induced by seed priming with salicylic acid. Journal of Agronomy and Crop Science 194, 161–168.
  21. Goklany S, Loring R.H, Glick J, Lee-Parsons C.W. 2009. Assessing the limitations to terpenoid indole alkaloid biosynthesis in Catharanthus roseus hairy root cultures through gene expression profiling and precursor feeding. Biotechnology Progress 25, 1289–1296.
  22. Hayat Q, Hayat S, Irfan M, Ahmad A. 2010. Effect of exogenous salicylic acid under changing environment: A review. Environmental and Experimental Botany 68, 14–25.
  23. Horváth E, Szalai G, Janda T. 2007. Induction of abiotic stress tolerance by salicylic acid signaling. Plant Growth Regulation 26, 290-300.
  24. Janda T, Szalai G, Tari I, Paldi E. 1999. Hydroponic treatment with salicylic acid decreases the effects of chilling injury in maize (Zea mays L.) plants. Planta 208, 175-180.
  25. Janda T, Szalai G, Lesko K, Yordanova R, Apostol S, Popova L. 2007. Factors contributing to enhanced freezing tolerance in wheat during frost hardening in the light. Phytochemistry 68, 1674–1782.
  26. Kamenetsky R, Barzilay A, Erez A, Halevy A.H. 2003. Temperature requirements for floral development of Herbaceous paeony cv. “Sarah Bernhardt”. Scientia Horticulturae 97, 309-320.
  27. Kar M, Mishra D. 1976. Catalase, peroxidase, and polyphenoloxidase activities during rice leaf senescence. Plant Physiology 57, 315-319.
  28. Koornneef M, Bentsink L, Hilhorst H. 2002. Seed dormancy and germination. Current Opinion in Plant Biology 5, 33-36.
  29. Körner C, Basler D. 2010. Phenology under global warming. Science 327, 1461–1462.
  30. Kuk Y.I, San Shin J, Burgos N.R, Tay Eak Hwang O, Baik Ho Cho, Sunyo Jung, Ja Ock G. 2003. Antioxidative enzymes offer protection from chilling damage in rice plants. Crop Science 43, 2109-2117.
  31. Kumar P, Lakshmi N.J, Mani V.P. 2000. Interactive effects of salicylic acid and phytohormones on photosynthesis and grain yield of soybean (Glycine max L. Merrill). Physiology and Molecular Biology of Plants 6, 186-179.
  32. Laube J, Sparks T.H, Estrella N, Höffler J, Ankerst P.D, Menzel A. 2014. Chilling outweighs photoperiod in preventing precocious spring development. Global Change Biology 20, 170–182.
  33. Lee S.S, Kim J.H. 2000. Total sugars, α-amylase activity, and emergence after priming of normal and aged rice seeds. Korean Journal of Crop Science 45, 108–111.
  34. Lukatkin A.S. 2002. Contribution of oxidative stress to the development of cold-Induced damage to leaves of chilling-sensitive plants: The activity of antioxidant enzymes during Plant chilling. Russian Journal of Plant Physiology 49, 878-885.
  35. Maguire J.D. 1962. Speed of germination - aid in selection and evaluation for seedling emergence and vigor. Crop Science 2, 176-177.
  36. Martin-Mex R, Villanueva-Couoh E, Herrera-Campos T, arqué-Saavedra A. 2001. Positive effect of salicylates on the flowering of african violet. Scientia Horticulturae 103, 499-502.
  37. Mutlu S, Atici O, Nalbantoglu B. 2009. Effects of salicylic acid and salinity on apoplastic antioxidant enzymes in two wheat cultivars differing in salt tolerance. Russian Journal of Plant Physiology 34, 334-338.
  38. Nayyar H, Bains T.S, Sanjeev K. 2005. Chilling stressed chickpea seedlings: Effect of cold acclimation, calcium and abscisic acid on cryoprotective solutes and oxidative damage. Environmental and Experimental Botany 54, 275-285.
  39. O´Kane D, Gill V, Boyd P, Burdun R. 1996. Chilling, oxidative stress and antioxidant responses in Arabidopsis thaliana callus. Planta 198, 371-377.
  40. Polgar C, Gallinat A, Primack R.B. 2014. Drivers of leaf-out phenology and their Implications for species Invasions: Insights from Thoreau’s Concord. New Phytologist 202, 106–115.
  41. Rajjou L, Belghazi M, Huguet R, Robin C, Moreau A, Job C, Job D. 2006. Proteomic Investigation of the effect of salicylic acid on arabidopsis seed germination and establishment of early defense mechanisms. Plant Physiology 141, 910-923.
  42. Reid KE, Niclas O, James S, Fred P, Steven TL. 2006. An optimized grapevine RNA Isolation procedure and statistical determination of reference genes for Real-Time RT-PCR during berry development. BMC Plant Biology 6, 27–35.
  43. Roh M.S. 2005. Flowering and Inflorescence development of Lachenalia aloides “Pearsonii” as Influenced by bulb storage and forcing temperature. Scientia Horticulturae 104, 305-323.
  44. Senaratna T, Touchell D, Bunn E, Dixon K. 2000. Acetyl salicylic acid (Aspirin) and salicylic acid induce multiple stress tolerance in bean and tomato plants. Plant Growth Regulation 30, 157-161.
  45. Srivalli B, Sharma G, Khanna-Chopra R. 2003. Antioxidative defense system in an upland rice cultivar subjected to increasing intensity of water stress following by recovery. Physiologia Plantarum 119, 503-512.
  46. Thomashow M.F. 1999. Plant cold tolerance: Freezing tolerance genes and regulatory mechanisms. Annual Review of Plant Physiology and Plant Molecular Biology 50, 571-599.
  47. Toole E.H, Hendricks S.B, Borthwick H.A, Toole V.K. 1956. Physiology of seed germination. Annual Review of Plant Physiology 7, 299-324.
  48. Van der Heijden R, Jacobs D.I, Snoeijer W. 2004. The Catharanthus alkaloids: pharmacognosy and biotechnology. Current Medicinal Chemistry 11, 1241-1253.
  49. Verma S, Dubey R.S. 2003. Lead toxicity induces lipid peroxidation and alters the activities of antioxidant enzymes in growing rice plants. Plant Science 164, 645-655.
  50. Xie Z.h, Zhang Zh-L, Honzlik Sh, Cook E, Shen Q.J. 2007. Salicylic acid inhibits gibberellin-induced alpha-amylase expression and seed germination via a pathway involving an abscisic-acid-inducible WRKY gene. Plant Molecular Biology 64, 293-303.      
  51. Xu R.Y, Nimi Y, Kojima K. 2007. Exogenous GA3 overcomes bud deterioration in tulip (Tulipagesneriana L.) bulbs during dry storage by promoting endogenous IAA activity in the internodes. Plant Growth Regulation 52, 1-8.
  52. Yong Z, Hao-Ru T, Ya L. 2008. Variation in antioxidant enzyme activities of two strawberry cultivars with short-term low temperature stress. World Journal of Agricultural Sciences 4, 458-462.
  53. Yordanova R.Y, Popova L.P. 2007. Effect of exogenous treatment with salicylic acid on photosynthetic activity and antioxidant capacity of chilled wheat plants. General and Applied Plant Physiology 33, 155-170.