The effects of heat stress (+40 °C for 2 hours), chilling (+4 °C for 2 hours), and moderate soil drought (four days without watering) on growth and cytokinin homeostasis in rye at 14th and 18th days of growth, and after recovery on 21st day were investigated. Morphometric analysis revealed that shoots were more sensitive to stresses, however, the root system recovered better in terms of fresh and dry weight. Cytokinin dynamics and distribution were organ-specific and stress-dependent. All stresses led to the accumulation of cytokinins in shoots and a decrease in roots, with the most significant changes observed under drought conditions. Stresses had prolonged effects on cytokinin levels, which remained lower than controls after recovery. The most significant alterations were found in trans-zeatinriboside accumulation, suggesting its potential role in adaptation to stress in rye. These findings provide a basis for further research into stress-resistance mechanisms in cereals, and the development of strategies to enhance their adaptability, in particular through the use of exogenous phytohormones.
Keywords: Secale cereale L., chilling, cytokinins, heat stress, growth, soil drought, stress resistance
Full text and supplemented materials
Free full text: PDFReferences
1. Karamushka, V., Boychenko, S., Kuchma, T. & Zabarna, O. (2022). Trends in the environmental conditions, climate change and human health in the southern region of Ukraine. Sustainability, 14, No. 9, 5664. https://doi.org/10.3390/su14095664
2. Vyshnevskyi, V.I. (2025). Climate change in Ukraine and its consequences. J. Landscape Ecol., 18, No. 4, pp. 150-174. https://doi.org/10.2478/jlecol-2025-0032
3. Narayanan, S. (2018). Effects of high temperature stress and traits associated with tolerance in wheat. Open Access J. Sci., 2, No. 3, pp. 177-186. https://doi.org/10.15406/oajs.2018.02.00067
4. Prasad, P.V.V., Pisipati, S.R., Mutava, R.N. & Tuinstra, M.R. (2008). Sensitivity of grain sorghum to high temperature stress during reproductive development. Crop Sci., 48, No. 5, pp. 1911-1917. https://doi.org/10.2135/cropsci2008.01.0036
5. Ji, H., Xiao, L., Xia, Y., Song, H., Liu, B., Tang, L., Cao, W., Zhu, Y. & Liu, L. (2017). Effects of jointing and booting low temperature stresses on grain yield and yield components in wheat. Agricult. Forest Meteorol., 243, pp. 33-42. https://doi.org/10.1016/j.agrformet.2017.04.016
6. Wu, Y., Wang, X., Peng, X., Ge, J., Cai, J., Huang, M., Zhou, O., Zhong, Y. & Jiang, D. (2023). Cold priming improves chilling resistance in wheat seedlings: Changing of photosystem II imprints during recovery from priming. Environ. Exp. Bot., 207, No. 1, 105220. https://doi.org/10.1016/j.envexpbot.2023.105220
7. Hu, W., Ren, T., Meng, F., Cong, R., Li, X., White, P.J & Lu, J. (2019). Leaf photosynthetic capacity is regulated by the interaction of nitrogen and potassium through coordination of CO2 diffusion and carboxylation. Physiol. Plant., 167, No. 3, pp. 418-432. https://doi.org/10.1111/ppl.12919
8. Lind, C., Dreyer, I., LЩpez-Sanjurjo, E.J., von Meyer, K., Ishizaki, K., Kohchi, T., Lang, D., Zhao, Y., Kreizer, I., Al-Rasheid, K.A.S., Ronne, H., Reski, R., Zhu, J-K., Geiger, D. & Hedrich, R. (2015). Stomatal guard cells co-opted an ancient ABA-dependent desiccation survival system to regulate stomatal closure. Current Biol., 25, No. 7, pp. 928-935. https://doi.org/10.1016/j.cub.2015.01.067
9. Dietrich, D. (2018). Hydrotropism: how roots search for water. J. Exp. Bot., 69, No. 11, pp. 2759-2771. https://doi.org/10.1093/jxb/ery034
10. Bushuk, W. (2001). Rye: Production, Chemistry, and Technology (2nd ed). St Paul, MN: AACC International, Inc. 239 p.
11. Arendt, E.K. & Zannini, E. (2013). 6 - Rye. Editor(s): Elke K. Arendt, Emanuele Zannini. In Woodhead Publishing Series in Food Science, Technology and Nutrition. Cereal Grains for the Food and Beverage Industries, Woodhead Publishing: 220-243e. https://doi.org/10.1533/9780857098924.220
12. Symonenko N.V., Skoryk V.V., Zhemoida, V. (2018). Results of the breeding work with winter rye to the Nosivska breeding research station. Plant Soil Sci., 268 [in Ukrainian]. https://agriculturalscience.com.ua/web/uploads/pdf/10847-23510-1-SM.pdf
13. Kosakivska, I.V., Babenko, L.M., Vasyuk, V.A., Voytenko, L.V. & Shcherbatiuk M.M. (2024). Chapter 2. Natural growth regulators as inducers of resistance in cereal plants against extreme environmental factors. In T.O. Yastreb, Yu.E. Kolupaev, A.I. Yemets, Ya.B. Blume (Eds.). Regulation of adaptive responses in plants. Published by Nova Science Publishers, Inc., New York: pp. 33-82. https://doi.org/10.52305/TXQB2084
14. Kosakivska, I.V., Shcherbatiuk, M.M., Vasyuk, V.A. & Voytenko, L.V. (2024 b). Phytohormones in growth regulation and the formation of stress resistance in cultivated cereals. Fiziol. rast. genet., 56, No. 2, pp. 130-150 [in Ukrainian]. https://doi.org/10.15407/frg2024.02.130
15. Panozzo, A., Bolla, P.K., Barion, G., Botton, A. & Vamerali, T. (2025). Phytohormonal regulation of abiotic stress tolerance, leaf senescence and yield response in field crops: A comprehensive review. BioTech, 14, No. 1, 14. https://doi.org/10.3390/biotech14010014
16. Vedenicheva, N.P. & Kosakivska, I.V. (2017). Cytokinins as regulators of plant ontogenesis under different growth conditions. Kyiv: Institute of Botany NAS of Ukraine, Nash Format, 200 p. [in Ukrainian]. https://www.botany.kiev.ua/doc/kos_ved.pdf
17. Vedenicheva, N.P. & Kosakivska, I.V. (2020). Cytokinins in cereals ontogenesis and adaptation. Fiziol. rast. genet., 52, No. 1, pp. 3-30 [in Ukrainian]. https://doi.org/10.15407/frg2020.01.003
18. da Cunha Neto, A.R., dos Santos AmbrЩsio, A., de Jesus Rodrigues Resende, A., Rѕgis Santos, B. & Nadal, M.C. (2025). From cell division to stress tolerance: the versatile roles of cytokinins in plants. Phyton-Int. J. Exp. Bot., 94, No. 3, pp. 539-560. https://doi.org/10.32604/phyton.2025.061776
19. Vedenicheva, N.P. & Kosakivska, I.V. (2016). Modern aspects of cytokinins studies: evolution and crosstalk with other phytohormones. Fiziol. rast. genet., 48, No. 1, pp. 3-19 [in Ukrainian]. https://doi.org/10.15407/frg2016.01.003
20. Mughal, N., Shoaib, N., Chen, J., Li, Y., He, Y., Fu, M., Li, X., He, Y., Guo, J., Deng, J., Yang, W. & Liu, J. (2024). Adaptive roles of cytokinins in enhancing plant resilience and yield against environmental stressors. Chemosphere, 364, 143189. https://doi.org/10.1016/j.chemosphere.2024.143189
21. Prasad, R. (2022). Cytokinin and its key role to enrich the plant nutrients and growth under adverse conditions-an update. Front. Genet., 13, 883924. https://doi.org/10.3389/fgene.2022.883924
22. Ullah, I., Toor, M.D., Basit, A. & Mohamed, H.I. (2024). Cytokinin Signaling in Plant Response to Abiotic Stresses. Abd-Elsalam, K.A., & Mohamed, H.I. (Eds.). Plant Growth Regulators to Manage Biotic and Abiotic Stress in Agroecosystems (1st ed.). CRC Press, pp. 26. https://doi.org/10.1201/9781003389507
23. Li, S.M., Zheng, H.X., Zhang, X.S. & Sui, N. (2021). Cytokinins as central regulators during plant growth and stress response. Plant Cell Rep., 40, No. 2, pp. 271-282. https://doi.org/10.1007/s00299-020-02612-1
24. Vedenicheva, N. & Kosakivska, I. (2024). Chapter. The regulatory role of cytokinins in adaptive responses of cereal plants. In book: Regulation of adaptive responses in plants, Nova Science Publication, USA, pp. 83-110. https://doi.org/10.52305/TXQB2084
25. Karunadasa, S., Kurepa, J. & Smalle, J.A. (2022). Gain-of-function of the cytokinin response activator ARR1 increases heat shock tolerance in Arabidopsis thaliana. Plant Signal. Behav., 17, No. 1, 2073108. https://doi.org/10.1080/15592324.2022.2073108
26. Xing, Y.H., Lu, H., Zhu, X., Deng, Y., Xie, Y., Luo, Q. & Yu, J. (2024). How rice responds to temperature changes and defeats heat stress. Rice, 17, p. 73. https://doi.org/10.1186/s12284-024-00748-2
27. Bhattacharya, A. (2022). Plant growth hormones in plants under low-temperature stress: A review. In: Physiological Processes in Plants Under Low Temperature Stress. Springer, Singapore. https://doi.org/10.1007/978-981-16-9037-2_6
28. Zheng, X.W., Cao, X.Y., Jiang, W.H., Xu, G.Z., Liang, Q.Z. & Yang, Z.Y. (2024). Cryoprotectant-mediated cold stress mitigation in litchi flower development: transcriptomic and metabolomic perspectives. Metabolites, 14, No. 4, p. 223. https://doi.org/10.3390/metabo14040223
29. Kaur, G. & Mishra, D. (2022). AtABCG14: a long-distance root-to-shoot carrier of cytokinin. Int. J. Plant Biol., 13, No. 3, pp. 352-355. https://doi.org/10.3390/ijpb13030029
30. Sakakibara, H. (2021). Cytokinin biosynthesis and transport for systemic nitrogen signaling. Plant J., 105, No. 2, pp. 421-430. https://doi.org/10.1111/tpj.15011
31. Bielach, A., Hrtyan, M. & Tognetti, V.B. (2017). Plants under stress: involvement of auxin and cytokinin. Int. J. Mol. Sci., 18, No. 7, p. 1427. https://doi.org/10.3390/ijms18071427
32. Wang, X-L., Qin, R-R., Sun, R-H., Wang, J-J., Hou, X-G., Qi, L., Shi, J., Li, X-L., Zhang, Y-F., Dong, P-H., Zhang, L-X. & Qin, D-H. (2018). No post-drought compensatory growth of corns with root cutting based on cytokinin induced by roots. Agricult. Water Manag., 205, No. C, pp. 9-20. https://doi.org/10.1016/j.agwat.2018.04.035
33. Werner, T., Nehnevajova, E., KШllmer, I., Nov«k, O., Strnad, M., Kr¬mer, U. & Schmтllinget, T. (2010). Root-specific reduction of cytokinin causes enhanced root growth, drought tolerance, and leaf mineral enrichment in Arabidopsis and tobacco. Plant Cell, 22, No. 12, pp. 3905-3920. https://doi.org/10.1105/tpc.109.072694
34. Kosakivska, I.V., Voytenko, L.V., Vasyuk, V.A. & Shcherbatiuk, M.M. (2024 c). ABA-induced alterations in cytokinin homeostasis of Triticum aestivum and Triticum speltaunder heat stress. Plant Stress, 11, p. 100353. https://doi.org/10.1016/j.stress.2024.100353
35. Yamamuro, C., Zhu, J.K. & Yang, Z. (2016). Epigenetic modifications and plant hormone action. Mol. Plant, 9, No. 1, pp. 57-70. https://doi.org/10.1016/j.molp.2015.10.008
36. Kosakivska, I.V., Vasyuk, V.A., Voytenko, L.V. & Shcherbatiuk, M.M. (2022). Changes in hormonal status of winter wheat (Triticum aestivum L.) and spelt wheat (Triticum spelta L.) after heat stress and in recovery period. Cereal Res. Commun., 50, pp. 821-830. https://doi.org/10.1007/s42976-021-00206-5
37. Kosakivska, I.V., Vedenicheva, N.P., Babenko, L.M., Voytenko, L.V., Romanenko, K.O. & Vasyuk, V.A. (2022). Exogenous phytohormones in the regulation of growth and development of cereals under abiotic stresses. Mol. Biol. Rep., 49, No. 1, pp. 617-628. https://doi.org/10.1007/s11033-021-06802-2
38. Kosakivska, I.V., Vasyuk, V.A., Voytenko, L.V. & Shcherbatiuk, M.M. 2023. The effects of moderate soil drought on phytohormonal balance of Triticum aestivum L. and Triticum spelta L. Cereal Res. Commun., 51, pp. 627-638. https://doi.org/10.1007/s42976-022-00332-8
39. Vedenicheva, N., Futorna, O., Shcherbatyuk, M. & Kosakivska, I. (2022). Effect of seed priming with zeatin on Secale cereale L. growth and cytokinins homeostasis under hyperthermia. J. Crop Improv., 36, No. 5, pp. 656-674. https://doi.org/10.1080/15427528.2021.2000909
40. Kosakivska, I.V., Shcherbatiuk, M.M. & Voytenko, L.V. (2020). Profiling of hormones in plant tissues: history, modern approaches, use in biotechnology. Biotechnol. Acta, 13, No. 4, pp. 14-25. https://doi.org/10.15407/biotech13.04.014
41. Shcherbatiuk, M.M., Voytenko, L.V., Kharkhota, M.A. & Kosakivska, I.V. (2021). Profiling of cytokinins in plant tissues: sampling, qualitative and quantitative analysis. Fiziol. rast. genet., 53, No. 4, pp. 346-368 [in Ukrainian]. https://doi.org/10.15407/frg2021.04.346
42. Cortleven, A., Leuendorf, J.E., Frank, M., Pezzetta, D., Bolt, S. & Schmтlling, T. (2019). Cytokinin action in response to abiotic and biotic stresses in plants. Plant Cell Environ., 42, pp. 998-1018. https://doi.org/10.1111/pce.13494
43. Sosnowski, J., Truba, M. & Vasileva, V. (2023). The impact of auxin and cytokinin on the growth and development of selected crops. Agriculture, 13, No. 3, p. 724. https://doi.org/10.3390/agriculture13030724
44. Svolacchia, N. & Sabatini, S. (2023). Cytokinins. Current Biol., 33(1): R10-R13. https://doi.org/10.1016/j.cub.2022.11.022
45. Voytenko, L.V., Shcherbatiuk, M.M., Vasyuk, V.A. & Kosakivska, I.V. (2024). Role of cytokinins in the regulation the chilling stress response in Triticum aestivum and Triticum spelta. Dopov. Nac. akad. nauk Ukr., No. 3, pp. 69-76. https://doi.org/10.15407/dopovidi2024.03.069
46. Thu, N.B.A., Hoang, X.L.T., Truc, M.T., Sulieman, S., Thao, N.P. & Tran, L.S.P. (2017). Cytokinin signaling in plant response to abiotic stresses. In: Pandey G.K., ed. Mechanism of Plant Hormone Signaling Under Stress. Hoboken: John Wiley & Sons, Inc., pp. 71-100. https://doi.org/10.1002/9781118889022.ch4
47. Feng, Y., Li, Z., Kong, X., Khan, A., Ullah, N. & Zhang, X. (2025). Plant coping with cold stress: molecular and physiological adaptive mechanisms with future perspectives. Cells, 14, No. 2, 110. https://doi.org/10.3390/cells14020110
48. Kopeck«, R., Kameniarov«, M., ‡ernъ, M., Brzobohatъ, B. & Nov«k, J. (2023). Abiotic stress in crop production. Int. J. Mol. Sci., 24, No. 7, 6603. https://doi.org/10.3390/ijms24076603
49. Feller, U. & Vaseva, I.I. (2014). Extreme climatic events: impacts of drought and high temperature on physiological processes in agronomically important plants. Front. Environ. Sci., 2, 39. https://doi.org/10.3389/fenvs.2014.00039
50. Liu, X. & Huang, B. (2002). Cytokinin effects on creeping bentgrass response to heat stress: II. Leaf senescence and oxidant metabolism. Crop Sci., 42, pp. 466-472. https://doi.org/10.2135/cropsci2002.0466
51. Todorova, D., Genkov, T., Vaseva-Gemisheva, I., Alexiva, V., Karanov, E., Smith, A. & Hall, M. (2005). Effect of temperature stress on the endogenous cytokinin content in Arabidopsis thaliana (L.) Heynh plants. Acta Physiol. Plant., 27, pp. 13-18. https://doi.org/10.1007/s11738-005-0031-5
52. Caers, M., Rudelsheim, P. & Van Onckelen, H. (1985). Effect of heat stress on photosynthetic activity and chloroplast infrastructure in correlation with endogenous cytokinin concentration in maize seedlings. Plant Cell Physiol., 26, pp. 47-52.
53. Xu, Y., Tian, J., Gianfagna, T. & Huang, B. (2009). Effects of SAG12-ipt expression on cytokinin production, growth and senescence of creeping bentgrass (Agrostis stolonifera L.) under heat stress. Plant Growth Regul., 57, pp. 281-291. https://doi.org/10.1007/s10725-008-9346-8
54. Kalapos, B., Nov«k, A., Dobrev, P., VHt«mv«s, P., Marincs, F., Galiba, G. & Vankov«, R. (2017). Effect of the winter wheat cheyenne 5A substituted chromosome on dynamics of abscisic acid and cytokinins in freezing-sensitive chinese spring genetic background. Front. Plant Sci., 8, 2033. https://doi.org/10.3389/fpls.2017.02033
55. Prerostova, S., Dobrev, P.I., Gaudinova, A., Knirsch, V., KШrber, N., Pieruschka, R., Fiorani, F., Brzobohatъ, B., ‡ernъ, M., Spichal, L., Humplik J., Vanek T., Schurr U. & Vankova, R. (2018). Cytokinins: their impact on molecular and growth responses to drought stress and recovery in Arabidopsis. Front. Plant Sci., 9, 655. https://doi.org/10.3389/fpls.2018.00655
56. Romanenko, K., Babenko, L., Smirnov, O. & Kosakivska, I. (2022). Antioxidant protection system and photosynthetic pigment composition in Secale cerealesubjected to short-term temperature stresses. The Open Agricult. J., 16 (Suppl-1, M3), e187433152206273. https://doi.org/10.2174/18743315-v16-e2206273
57. Ramireddy, E., Hosseini, S.A., Eggert, K., Gillandt, S., Gnad, H., von Wirѕn, N. & Schmтlling, T. (2018). Root engineering in barley: increasing cytokinin degradation produces a larger root system, mineral enrichment in the shoot and improved drought tolerance. Plant Physiol., 177, No. 3, pp. 1078-1095. https://doi.org/10.1104/pp.18.00199
58. Vojta, P., Kok«л, F., Husi№kov«, A., Gryz, J., Bergougnoux, V., Marchetti, C.F., Jiskrov«, E., Jeыilov«, E., Mik, V., Ikeda, Y. & Galuszka, P. (2016). Whole transcriptome analysis of transgenic barley with altered cytokinin homeostasis and increased tolerance to drought stress. New Biotechnol., 33, (5 Pt B), pp. 676-691. https://doi.org/10.1016/j.nbt.2016.01.010
59. PospHлilov«, H., Jiskrov«, E., Vojta, P., MrHzov«, K., Kok«л, F., ‡udejkov«, M.M., Bergougnoux, V., PlHhal, O., Klimeлov«, J., Nov«k, O., Dzurov«, L., Frѕbort, I. & Galuszka, P. (2016). Transgenic barley overexpressing a cytokinin dehydrogenase gene shows greater tolerance to drought stress. New Biotechnol., 33, Part (B), pp. 692-705. https://doi.org/10.1016/j.nbt.2015.12.005
60. Oneto, C.D., Otegui, M.E., Baroli, I., Beznec, A., Faccio, P., Bossio, E., Blumwald, E. & Lewi, D. (2016). Water deficit stress tolerance in maize conferred by expression of an isopentenyltransferase (IPT) gene driven by a stress- and maturation-induced promoter. J. Biotechnol., 220, pp. 66-77. https://doi.org/10.1016/j.jbiotec.2016.01.014
61. Xu, Y., Burgess, P., Zhang, X. & Huang, B. (2016). Enhancing cytokinin synthesis by overexpressing ipt alleviated drought inhibition of root growth through activating ROS-scavenging systems in Agrostis stolonifera. J. Exp. Bot., 67, No. 6, pp. 1979-1992. https://doi.org/10.1093/jxb/erw019