Visn. Hark. nac. agrar. univ., Ser. Biol., 2020, Issue 2 (50), p. 93-104


O. I. Horielova1, M. A. Shkliarevskyi1, N. I. Ryabchun2, L. F. Kabashnikova3, Yu. E. Kolupaev1, 4

1Dokuchaev Kharkiv National Agrarian University
(Kharkiv, Ukraine)


2Yurjev Рlant Production Institute
of National Academy of Agrarian Sciences of Ukraine
(Kharkiv, Ukraine)
3Institute of Biophysics and Cell Engineering
of National Academy of Sciences of Belarus
(Minsk, Belarus)
4Karazin Kharkiv National University
(Kharkiv, Ukraine)

It is known that salicylic acid and nitric oxide (NO) are involved in the formation of many adaptive reactions of plants to action of stressors of various natures. There is data that treatment of plants with salicylic acid and nitric oxide donors increases their resistance to low positive temperatures. At the same time, their effect on the resistance of plants to cryostress has been studied deficiently. It is known that some physiological effects of salicylic acid are realized with the participation of NO as a signaling mediator. However, the combined effect of salicylic acid and NO donors on physiological processes responsible for the formation of frost resistance has not been studied yet. We investigate the influence of priming of wheat seed (Triticum aestivum L.) with salicylic acid and NO donor – sodium nitroprusside (SNP) – separately and together on the formation of frost resistance of etiolated wheat seedlings during hardening at 2-4°С. The increase of the seedlings survival after freezing at –6 and –8°С under the influence of salicylic acid and SNP has been shown. The protective effect of the mutual treatment of wheat seeds with salicylic acid (10 μM) and SNP (100 μM) has been more noticeable. Cold hardening of seedlings, as well as the preliminary treatment of seeds with salicylic acid, caused an increase in the activity of superoxide dismutase (SOD), catalase and guaiacol peroxidase, the content of proline and sugars in the seedlings’ tissues. The priming of seed with SNP contributed to an increase in the SOD and catalase activity in wheat seedlings, and in the proline and sugars content. An additional increase in SOD activity and sugars content in seedlings with the combined use of salicylic acid and SNP has been noted. Possible causes of the enhancement of stress-protective effects of salicylic acid and NO donor with their combined action has been discussed.

Key words: Triticum aestivum, salicylic acid, nitric oxide, frost resistance, antioxidant system, osmolytes



1. Evdokimova O.V., Kabashnikova L.F., Savchenko G.E. 2014. Salicylic acid and reactive oxygen species content in leaves barley (Hordeum vulgare) at salicylates treatments. Proceedings of the National Academy of Sciences of Belarus, Biological Series. 3 : 57-62. (In Russian).
2. Karpets Yu.V., Kolupaev Yu.E., Kosakivska I.V. 2016. Nitric oxide and hydrogen peroxide as signal media-tors at induction of heat resistance of wheat plantlets by exogenous jasmonic and salicylic acids. Fiziol. rast. genet. 48 (2) :158-166. (In Russian).
3. Kolupaev Yu.E., Karpets Yu.V., Yastreb T.O., Lugovaya A.A. 2018. Combined effect of salicylic acid and nitrogen oxide donor on stress-protective system of wheat plants under drought conditions. Appl. Biochem. Microbiol. 54 (4) : 418-424.
4. Kolupaev Yu.E., Ryabchun N.I., Vayner A.A., Yastreb T.O., Oboznyi A.I. 2015. Antioxidant enzyme activity and osmolyte content in winter cereal seedlings under hardening and cryostress. Russ. J. Plant Physiol. 62 (4) : 499-506.
5. Pashkevich L.V., Kabashnikova L.F. 2018. Role of sali-cylic acid in formation of system acquired resistance of plants at pathogenesis. Visn. Hark. nac. agrar. univ., Ser. Biol. 3 (45) : 31-48. (In Russian).
6. Kholoptseva E.S., Ignatenko A.A., Repkina N.S., Talanova V.V. 2019. Characteristics of wheat plant re-sponses to short-term and prolonged exposure to salicylic acid under optimal and low temperature. Trudy' Karel'skogo nauchnogo czentra RAN. 12: 19-30. (In Russian).
7. 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. Biol. Plant. 49 : 541-550.
8. Alavi S.M.N., Arvin M.J., Kalantari K.M. Salicylic acid and nitric oxide alleviate osmotic stress in wheat (Triticum aestivum L.) seedlings. J. Plant Interact. 2014. 9 (1) : 683-688.
9. Arora D., Jain P., Singh N., Kaur H., Bhatla S.C. 2016. Mechanisms of nitric oxide crosstalk with reactive oxygen species scavenging enzymes during abiotic stress tolerance in plants. Free Radical Res. 50 : 291-303.
10. Bates L.S., Walden R.P., Tear G.D. 1973. Rapid determination of free proline for water stress studies. Plant Soil. 39 : 205-210.
11. Baudouin E.,Jeandroz S. 2015. Nitric oxide as a media-tor of cold stress response: a transcriptional point of view. In: Nitric Oxide Action in Abiotic Stress Re-sponses in Plants. Eds. Khan M.N. et al. Switzerland : Springer International Publishing : 129-139.
12. Esim N., Atici Ö. 2015. Effects of exogenous nitric oxide and salicylic acid on chilling-induced oxidative stress in wheat (Triticum aestivum). Front. Life Sci. 8 (2) : 124-130.
13. Fan J., Chen K., Amombo E., Hu Z., Chen L., Fu J. 2015. Physiological and molecular mechanism of nitric oxide (NO) involved in bermudagrass response to cold stress. Plos ONE.
14. Fancy N.N., Bahlmann A.K., Loake G.J. 2017. Nitric oxide function in plant abiotic stress. Plant Cell En-viron. 40 (4) : 462-472.
15. 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. J. Agron.Crop Sci. 194 : 161-168.
16. Hamayun M., Khan A.L., Ahmad N., Lee I.J., Khan S.A.;, Shinwari Z.K. 2010. Effect of polyethylene glycol induced drought stress on physiohormonal attributes of soybean. Pak. J. Bot. 42 (2) : 977-986.
17. Hashempour A., Ghasemnezhad M., Fotouhi Ghazvini R., Sohani M.M. 2014. The physiological and biochemical responses to freezing stress of olive plants treated with salicylic acid. Russ. J. Plant Physiol. 61 (4) : 443-450.
18. Horvath E., Szalai G., Janda T. 2007. Induction of abi-otic stress tolerance by salicylic acid signaling. J. Plant Growth Regul. 26 : 290-300.
19. Kim Y.S., Park S., Gilmour S.J., Thomashow M.F. 2013. Roles of CAMTA transcription factors and salicylic acid in configuring the low-temperature transcriptome and freezing tolerance of Arabidopsis. Plant J. 75 : 364-376.
20. Majláth I., Szalai G., Janda T. 2011. Exploration of cold signalling related to ascorbate and salicylic acid in Arabidopsis thaliana. Acta Biol. Szeged. 55 (1) : 117-118.
21. Min K., Showman L., Perera A., Arora R. 2018. Salicylic acid-induced freezing tolerance in spinach (Spinacia oleracea L.) leaves explored through metabolite profiling. Environ. Exp. Bot. 156 : 214-227.
22. Miura K., Tada Y. 2014. Regulation of water, salinity, and cold stress responses by salicylic acid. Front. Plant Sci. 5 : 4
23. Mostofa M.G., Fujita M., Tran, L.S.P. 2015. Nitric oxide mediates hydrogen peroxide- and salicylic acid-induced salt tolerance in rice (Oryza sativa L.) seed-lings. Plant Grow. Regul. 77 : 265-277.
24. Mutlu S., Karadağoğlu Ö., Atici Ö., Nalbantoğlu B. 2013a. Protective role of salicylic acid applied before cold stress on antioxidative system and protein patterns in barley apoplast. Biol. Plant. 57 (3) : 507-513.
25. Mutlu S., Karadağoğlu Ö., Atici Ö., Taşğin E., Nalbantoğlu B. 2013. Time-dependent effect of salicylic acid on alleviating cold damage in two barley cultivars differing in cold tolerance. Turk. J. Bot. 37 : 343-349.
26. Pal M., Gondor O.K., Janda T. 2013. Role of salicylic acid in acclimation to low temperature. Acta Agron. Hung. 61 (2) : 161-172.
27. Peleg-Grossman S., Melamed-Book N., Levine A. 2012. ROS production during symbiotic infection sup-presses pathogenesis-related gene expression. Plant Signaling Behav. 7. : 409-415.
28. Puyaubert J., Baudouin E. 2014. New clues for a cold case: nitric oxide response to low temperature. Plant Cell Environ. 37 : 2623-2630.
29. Shakirova F.M., Sakhabutdinova A.R., Bezruko-va M.V., Fatkhutdinova R., Fatkhutdinova D. 2003. Changes in the hormonal status of wheat seedlings induced by salicylic acid and salinity. Plant Sci. 164 : 317-322.
30. Shakirova F.M., Bezrukova M.V., Maslennikova D.R. 2013. Endogenous ABA as a hormonal intermediate in the salicylic acid induced protection of wheat plants against toxic. In: Salicylic Acid. Eds. Hayat S. et al. Dordrecht : Springer Science+Business Media : 119-140.
31. Shakirova F.M., Allagulova Ch.R., Maslennikova D.R., Klyuchnikova E.O., Avalbaev A.M., Bezrukova M.V. 2016. Salicylic acid-induced protection against cadmium toxicity in wheat plants. Environ. Exp. Bot. 122 : 19-28.
32. Shin H., Min K., Arora R. 2018. Exogenous salicylic acid improves freezing tolerance of spinach (Spina-cia oleracea L.) leaves. Cryobiology. 81 : 192-200.
33. Song F., Goodman R.M. 2001. Activity of nitric oxide is dependent on, but is partially required for function of, salicylic acid in the signaling pathway in tobacco systemic acquired resistance. Mol. Plant-Microbe Interact. 14 (12) : 1458-1462.
34. Tasgin E., Atici O., Nalbantoglu B. 2003. Effects of sal-icylic acid and cold on freezing tolerance in winter wheat leaves. Plant Growth Regul. 41 : 231-236.
35. Wang D.H., Li X.X., Su Z.K., Ren H.X. 2009. The role of salicylic acid in response of two rice cultivars to chilling stress. Biol. Plant. 53 (3) : 545-552,
36. Wang W., Wang X., Huang M., Cai J., Zhou Q., Dai T., Cao W., Jiang D. 2018. Hydrogen peroxide and ab-scisic acid mediate salicylic acid-induced freezing tolerance in wheat. Front. Plant Sci. 3 (9) : 1137.
37. Yemets A.I., Karpets Yu.V., Kolupaev Yu.E. Blume Ya.B. 2019. Emerging technologies for en-hancing ROS/RNS homeostasis. In: Reactive Oxygen, Nitrogen and Sulfur Species in Plants: Production, Metabolism, Signaling and Defense Mecha-nisms, V. 2. Eds. Hasanuzzaman M. et al.). John Wiley & Sons Ltd. : 873-922.
38. Yordanova R., Popova L. 2007. Effect of exogenous treatment with salicylic acid on photosynthetic activity and antioxidant capacity of chilled wheat plants. Gen. Appl. Plant Physiol. 33 (3-4) : 155-170.