Exogenous Ascorbic Acid Improves Defence Responses of Sunflower (Helianthus Annuus) Exposed to Multiple Stresses


Ascorbic acid is an important antioxidant that plays role both on growth and development and also stress response of the plant. The purpose of this study was to determine the effect of ascorbate on physiological and biochemical changes of sunflower that was exposed to multiple stresses. Chlorophyll and carotenoid contents decreased and glutathione, ascorbate and malondialdehyde contents as well as antioxidant enzyme activities increased for sunflower plant that was exposed to 50 mM NaCl and pendimethalin at different concentrations. These changes were found to be more significant in groups simultaneously exposed to both stress factors. While malondialdehyde content decreased, chlorophyll, carotenoid, ascorbate, glutathione contents and antioxidant enzyme activities increased in plants treated exogenously with ascorbate, compared to the untreated samples. According to the findings of our study; compared to individual stress, the effect of stress is more pronounced in sunflower exposed to multiple stresses, and treatment with exogenous ascorbate reduces the negative effects of stress.


  1. 1.

    Abbasi, M., Fakhani, E. (2015) Role of salicylic acid and ascorbic acid in the alleviation of salinity stres in wheat (Triticum aestivum L.. JBES 6, 107–113.

    Google Scholar 

  2. 2.

    Akerboom, T. P. M., Sies, H. (1981) Assay of glutathione, glutathione disulfide and glutathione mixed disulfide in biological samples, In: Jakoby, W. B. (ed.). Methods in Enzymology 77, Academic Press, New York, pp. 373–382.

    CAS  PubMed  Google Scholar 

  3. 3.

    Akram, N. A., Ashraf, M., Al-Qurainy, F. (2012) Aminolevulinic acid-induced changes in some key physiological attributes and activities of antioxidant enzymes in sunflower (Helianthus annuus L.) plants under saline regimes. Sci. Hortic. 142, 143–148.

    CAS  Google Scholar 

  4. 4.

    Andrews, C. J. (2005) Purifcation and characterisation of a family of glutathione transferases with roles in herbicide detoxifcation in soybean (Glycine max L.); selective enhancement by herbicides and herbicide safeners. Pestic. Biochem. Phys. 82, 205–219.

    CAS  Google Scholar 

  5. 5.

    Anjum, N. A., Gill, S. S., Gill, R., Hasanuzzaman, M., Duarte, A. C., Pereira, E., Ahmad, I., Tuteja, R., Tuteja, N. (2014) Metal/metalloid stress tolerance in plants: role of ascorbate, its redox couple, and associated enzymes. Protoplasma 251, 1265–1283.

    CAS  PubMed  Google Scholar 

  6. 6.

    Appleby, A., Valverde, B. (1989) Behavior of dinitroaniline herbicides in plants. Weed Technol. 3, 198–206.

    CAS  Google Scholar 

  7. 7.

    Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    CAS  Article  Google Scholar 

  8. 8.

    Bybordi, A. (2012) Effect of ascorbic acid and silicium on photosynthesis, antioxidant enzyme activity, and fatty acid contents in canola exposure to salt stress. JIA 11, 1610–1620.

    CAS  Google Scholar 

  9. 9.

    Carlberg, I., Mannervik, B. (1985) Glutathione reductase. Method. Enzymol. 113, 484–490.

    CAS  Google Scholar 

  10. 10.

    Çulha, Ş., Çakırlar, H. (2011) The effect of salinity on plants and salt tolerance mechanisms. AKU-J. Sci. Eng. 11, 11–34.

    Google Scholar 

  11. 11.

    De-Kok, L., Graham, M. (1980) Levels of pigments, soluble proteins, amino acids and sulfhydryl compounds in foliar tissue of Arabidopsis thaliana during dark induced and natural senescence. Plant Physiol. Biochem. 27, 133–142.

    Google Scholar 

  12. 12.

    Duncan, D. B. (1955) Multiple range and multiple F tests biometrics. IBS 11, 1–42.

    Google Scholar 

  13. 13.

    Ebrahimian, E., Bybordi, A. (2011) Influence of different proportion of nitrate, ammonium and silicium on activity of antioxidant enzymes and some physiological traits in sunflower under conditions of salt stress. JFAE 9, 1052–1058.

    CAS  Google Scholar 

  14. 14.

    Elloumi, N., Zouari, M., Chaari, L., Abdallah, F. B., Woodward, S., Kallel, M. (2015) Effect of phosphogypsum on growth, physiology, and the antioxidative defense system in sunflower seedlings. Environ. Sci. Pollut. 22, 14829–14840.

    CAS  Google Scholar 

  15. 15.

    Gill, S. S., Anjum, N. A., Hasanuzzaman, M., Gill, R., Trivedi, D. K., Ahmad, I., Pereira, E., Tuteja, N. (2013) Glutathione and glutathione reductase: A boon in disguise for plant abiotic stress defense operations. Plant Physiol. Biochem. 70, 204–212.

    CAS  PubMed  Google Scholar 

  16. 16.

    Gill, S. S., Tuteja, N. (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol. Biochem. 48, 909–930.

    CAS  Google Scholar 

  17. 17.

    Habig, W. H., Pabst, M. J., Jakoby, W. B. (1974) The first enzymatic step in mercapturic acid formation Glutathion S-Transferases. J. Biol. Chem. 249, 7130–7139.

    CAS  Google Scholar 

  18. 18.

    Haferkamp, M. R. (1988) Environmental factors affecting plant productivity. Achieving efficient use of rangeland resources. In: White, R. S., Short, R. E. (ed.). Fourth Keog Research Symposium. Montana Agr. Exp. Sta. Bozeman. p. 132.

    Google Scholar 

  19. 19.

    Heath, R. L., Packer, L. (1968) Photoperoxidation in isolated chloroplast, I. Kinetics stoichiometry of fatty acid peroxidation. Arch. Biochem. Biophys. 125, 180–198.

    Google Scholar 

  20. 20.

    Jabeen, N., Ahmad, R. (2013) The activity of antioxidant enzymes in response to salt stress in safflower (Carthamus tinctorius L.) and sunflower (Helianthus annuus L.) seedlings raised from seed treated with chitosan. J. Sci. Food Agr. 93, 1699–1705.

    CAS  Google Scholar 

  21. 21.

    Kalefetoğlu, T., Ekmekçi, Y. (2005) The effects of drought on plants and tolerance mechanisms. GUJS 18, 723–740.

    Google Scholar 

  22. 22.

    Kaya, A., Doganlar, Z. B. (2016) Exogenous jasmonic acid induces stress tolerance in tobacco (Nicotiana tabaccum) exposed to imazapic. Ecotox. Environ. Safe 124, 470–479.

    CAS  Google Scholar 

  23. 23.

    Kaya, A., Yiğit, E. (2014) The physiological and biochemical effects of salicylic acid on sunflowers (Helianthus annuus) exposed to flurochloridone. Ecotox. Environ. Safe 106, 232–238.

    CAS  Google Scholar 

  24. 24.

    Khan, A., Lang, I., Amjid, M., Shah, A., Ahmad, I., Nawaz, H. (2013) Inducing salt tolerance on growth and yield of sunflower by appliying different levels of ascorbic acid. J. Plant Nutr. 36, 1180–1190.

    CAS  Google Scholar 

  25. 25.

    Kostopoulou, Z., Therios, I., Roumeliotis, E., Kanellis, A. K., Molassiotis, A. (2015) Melatonin combined with ascorbic acid provides salt adaptation in Citrus aurantium L. seedlings. Plant Physiol. Biochem. 86, 155–165.

    CAS  PubMed  Google Scholar 

  26. 26.

    Li, G., Wan, S., Zhou, J., Yang, Z., Qin, P. (2010) Leaf chlorophyll fluorescence, hyperspectral reflectance, pigments content, malondialdehyde and proline accumulation responses of castor bean (Ricinus communis L.) seedlings to salt stress levels. Ind. Crop. Prod. 31, 13–19.

    Google Scholar 

  27. 27.

    Lichtenthaler, K., Welburn, A. R. (1983) Determination of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions, 603rd Meeting, Liverpool, pp. 591–592.

    Google Scholar 

  28. 28.

    Liu, J., Wang, W., Wang, L., Sun, Y. (2015) Exogenous melatonin improves seedling health index and drought tolerance in tomato. Plant Growth Regul. 77, 317–326.

    CAS  Google Scholar 

  29. 29.

    Mafakheri, A., Siosemardeh, A., Bahramnejad, B., Struik, P. C., Sohrabi, Y. (2010) Effect of drought stress on yield, proline and chlorophyll contents in three chickpea cultivars. AJCS 4, 580–585.

    CAS  Google Scholar 

  30. 30.

    Mandal, S., Yadav, S., Yadav, S., Nema, R. K. (2009) Antioxidants: A review. J. Chem. Pharm. Res 1, 102–104.

    Google Scholar 

  31. 31.

    Mitsou, K., Koulianou, A., Lambropoulou, D., Pappas, P., Albanis, T., Lekka, M. (2006) Growth rate effects, responses of antioxidant enzymes and metabolic fate of the herbicide propanil in the aquatic plant Lemna minor. Chemosphere 62, 275–284.

    CAS  PubMed  Google Scholar 

  32. 32.

    M.-Kalantari, K. H., Oloumi, H. (2005) Study the effects of CdCl2 on lipid peroxidation and antioxidant compounds content in Brassica napus. Iranian J. Sci. Technol. Trans. A. 29, 201–208.

    CAS  Google Scholar 

  33. 33.

    Mukherjee, S. P., Choudhuri, M. A. (1983) Implication of water stress induced changes in the levels of endogenous ascorbic acid and hydrogen peroxide in Vigna seedlings. Physiol. Plantarum 58, 166–170.

    CAS  Google Scholar 

  34. 34.

    Nakano, Y., Asada, K. (1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplasts. Plant Cell Physiol. 22, 867–880.

    CAS  Google Scholar 

  35. 35.

    Potters, G., Gara, L. D., Asard, H., Horemans, N. (2002) Ascorbate and glutathione: guardians of the cell cycle, partners in crime. Plant Physiol. Biochem. 40, 537–548.

    CAS  Google Scholar 

  36. 36.

    Sairam, R. K., Rao, K. V., Srivastava, G. C. (2002) Differential response of wheat genotypes to long term salinity stress in relation to oxidative stress, antioxidant activity and osmolyte concentration. Plant Sci. 163, 1037–1046.

    CAS  Google Scholar 

  37. 37.

    Santos, C. V. (2004) Regulation of chlorophyll biosynthesis and degradation by salt stress in sunflower leaves. Sci. Hortic. 103, 93–99.

    CAS  Google Scholar 

  38. 38.

    Sivaci, A., Kaya, A., Duman, S. (2014) Effects of ascorbic acid on some physiological changes of pepino (Solanum muriactum ait) under chilling stress. Acta Biol. Hung. 65, 305–318.

    CAS  PubMed  Google Scholar 

  39. 39.

    Shigeoka, S., Ishikawa, T., Tamoi, M., Miyagawa, Y., Takeda, T., Yabuta, Y., Yoshimura, K. (2002) Regulation and function of ascorbate peroxidase isoenzymes. J. Exp. Bot. 53, 1305–1319.

    CAS  Google Scholar 

  40. 40.

    Smirnoff, N. (1996) The function of metabolism of ascorbic acid in plants. Ann. Bot. London 78, 661–669.

    CAS  Google Scholar 

  41. 41.

    Sondhia, S. (2012) Dissipation of pendimethalin in soil and its residues in chickpea (Cicer arietinum 1.) under field conditions. B. Environ. Contam. Tox. 89, 1032–1036.

    CAS  Google Scholar 

  42. 42.

    Wang W., Vinocur, B., Altman, A. (2003) Plant responses to drought, salinity and extreme temperatures towards genetic engineering for stress tolerance. Planta 218, 1–14.

    CAS  PubMed  PubMed Central  Google Scholar 

  43. 43.

    Verma, K., Shekhawat, G. S., Sharma, A., Mehta, S. K., Sharma, V. (2008) Cadmium induced oxidative stress and changes in soluble and ionically bound cell wall peroxidase activities in roots of seedling and 3–4 leaf stage plants of Brassica juncea (L.) czern. Plant Cell Rep. 27, 1261–1269.

    CAS  PubMed  Google Scholar 

  44. 44.

    Zhang, J., Kirkham, M. B. (1996) Antioxidant responses to drought in sunflower and sorghum seedlings. New Phytol. 132, 361–373.

    CAS  PubMed  Google Scholar 

Download references

Author information



Corresponding author

Correspondence to Armagan Kaya.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kaya, A. Exogenous Ascorbic Acid Improves Defence Responses of Sunflower (Helianthus Annuus) Exposed to Multiple Stresses. BIOLOGIA FUTURA 68, 290–299 (2017). https://doi.org/10.1556/018.68.2017.3.6

Download citation


  • Ascorbate
  • sunflower
  • pendimethalin
  • NaCl
  • antioxidant