Effects of the Protein Hydrolysate Pretreatment on Cucumber Plants Exposed to Chilling Stress

Adelina Harizanova, Lyubka Koleva-Valkova, Andon Vassilev


This study aimed to evaluate the effects of the protein hydrolysate Naturamin WSP on the antioxidant defense system and oxidation-related damage of young cucumber plants exposed to chilling stress. Low positive temperatures have a negative effect on plant growth and performance, and besides visible alterations, such as inhibited growth, significant changes occur at the cellular level. Plants grown at low temperature typically suffer from oxidative damage, which leads to increased lipid peroxidation. Moreover, chilling-stressed plants accumulate more proline to protect their cell membranes. The application of biostimulants such as the protein hydrolysate Naturamin WSP can alleviate some of the adverse effects caused by low temperature. Our results indicated an increased activity of guaiacol peroxidase (GPOD) in all plants treated with the biostimulant regardless of the temperature of cultivation. The mitigation of damages caused by chilling stress might be explained by an enhanced anti-oxidative defense, as demonstrated by the activity of guaiacol peroxidases and increased proline concentrations in Naturamin WSP-treated plants.


antioxidative defense system; antioxidative enzyme; Cucumis sativus L.; low positive temperature stress; osmolytes; soluble protein

Full Text:



Bates, L. S., Waldren, R. P., & Teare, I. D. (1973). Rapid determination of free proline for water-stress studies. Plant and Soil, 39, 205–207. https://doi.org/10.1007/BF00018060

Bergmeyer, H., Gawehn, K., & Grassl, M. (1974). Enzymes as biochemical reagents. In H.-U. Bergmeier (Ed.), Methods in enzymatic analysis (2nd ed., Vol. 2, pp. 685–690). Academic Press.

Borowski, E. (2009). Response to chilling in cucumber (Cucumis sativus L.) plants treated with Triacontanol and Asahi SL. Acta Agrobotanica, 62(2), 165–172. https://doi.org/10.5586/aa.2009.038

Botta, A. (2013). Enhancing plant tolerance to temperature stress with amino acids: An approach to their mode of action. Acta Horticulturae, 1009, 29–35. https://doi.org/10.17660/ActaHortic.2013.1009.1

Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254. https://doi.org/10.1016/0003-2697(76)90527-3

Cholakova-Bimbalova, R., Petrov, V., & Vassilev, A. (2019). Photosynthetic performance of young maize (Zea mays L.) plants exposed to chilling stress can be improved by the application of protein hydrolysates. Acta Agrobotanica, 72(2), Article 1769. https://doi.org/10.5586/aa.1769

Colla, G., Hoagland, L., Ruzzi, M., Cardarelli, M., Bonini, P., Canaguier, R., & Rouphael, Y. (2017). Biostimulant action of protein hydrolysates: Unraveling their effects on plant physiology and microbiome. Frontiers in Plant Science, 8, Article 2202. https://doi.org/10.3389/fpls.2017.02202

Dumanović, J., Nepovimova, E., Natić, M., Kuča, K., & Jaćević, V. (2021). The significance of reactive oxygen species and antioxidant defense system in plants: A concise overview. Frontiers in Plant Science, 11, Article 552969. https://doi.org/10.3389/fpls.2020.552969

Ejaz, S., Fahad, S., Anjum, M., Nawaz, A., Naz, S., Hussain, S., & Ahmad, S. (2020). Role of osmolytes in the mechanisms of antioxidant defense of plants. In E. Lichtfouse (Ed.), Sustainable agriculture reviews (Vol. 39, pp. 95–117). Springer. https://doi.org/10.1007/978-3-030-38881-2_4

Ertani, A., Schiavon, M., Muscolo, A., & Nardi, S. (2013). Alfalfa plant-derived biostimulant stimulate short-term growth of salt stressed Zea mays L. plants. Plant Soil, 364, 145–158. https://doi.org/10.1007/s11104-012-1335-z

European Biostimulants Industry Council. (2019). Function defines biostimulant products. https://biostimulants.eu/issue/function-defines-biostimulant-products

Fu, X., Feng, Y.-Q., Zhang, X.-W., Zhang, Y.-Y., Bi, H.-G., & Ai, X.-Z. (2021). Salicylic acid is involved in rootstock–scion communication in improving the chilling tolerance of grafted cucumber. Frontiers in Plant Science, 12, Article 693344. https://doi.org/10.3389/fpls.2021.693344

Ghanbari, F., & Kordi, S. (2019). Hardening pretreatment by drought and low temperature enhanced chilling stress tolerance of cucumber seedlings. Acta Scientiarum Polonorum, Hortorum Cultus, 18(2), 29–37. https://doi.org/10.24326/asphc.2019.2.4

Gill, S., & Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 48, 909–930. https://doi.org/10.1016/j.plaphy.2010.08.016

Giri, J. (2011). Glycinebetaine and abiotic stress tolerance in plants. Plant Signaling & Behavior, 6(11), 1746–1751. https://doi.org/10.4161/psb.6.11.17801

Heath, R., & Packer, L. (1968). Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics, 125, 189–198. https://doi.org/10.1016/0003-9861(68)90654-1

Kuk, Y., & Shin, J. (2007). Mechanisms of low-temperature tolerance in cucumber leaves of various ages. Journal of the American Society for Horticultural Science, 132(3), 294–301. https://doi.org/10.21273/JASHS.132.3.294

Lukatkin, A. (2002). Contribution of oxidative stress to the development of cold-induced damage to leaves of chilling-sensitive plants: 2. The activity of antioxidant enzymes during plant chilling. Russian Journal of Plant Physiology, 49, 782–788. https://doi.org/10.1023/A:1020965629243

Lukatkin, A., Brazaityte, A., Bobinas, C., & Duchovski, P. (2012). Chilling injury in chilling-sensitive plants: A review. Žemdirbystė, 99(2), 111–124.

Nardi, S., Pizzeghello, D., Schiavon, M., & Ertani, A. (2016). Plant biostimulants: Physiological responses induced by protein hydrolyzed-based products and humic substances in plant metabolism. Scientia Agricola, 73(1), 18–23. https://doi.org/10.1590/0103-9016-2015-0006

Noctor, G., & Foyer, C. (1998). Ascorbate and glutathione: Keeping active oxygen under control. Annual Review of Plant Biology, 1, 249–279. https://doi.org/10.1146/annurev.arplant.49.1.249

Omoarelojie, L. O., Kulkarni, M. G., Finnie, G. F., & van Staden, J. (2021). Biostimulants and the modulation of plant antioxidant systems and properties. In S. Gupta & J. Van Staden (Eds.), Biostimulants for crops from seed germination to plant development. A practical approach (pp. 333–363). Academic Press. https://doi.org/jdf9

Pan, D.-Y., Fu, X., Zhang, X.-W., Liu, F.-J., Bi, H.-G., & Ai, X.-Z. (2020). Hydrogen sulfide is required for salicylic acid-induced chilling tolerance of cucumber seedlings. Protoplasma, 257, 1543–1557. https://doi.org/10.1007/s00709-020-01531-y

Phansak, P., Siriwong, S., Kanawapee, N., Thumanu, K., Gunnula, W., & Buensanteai, N. (2021). Drought response of rice in northeastern Thailand assessed via Fourier transform infrared spectroscopy. Acta Agrobotanica, 74, Article 7421. https://doi.org/10.5586/aa.7421

Ricci, M., Tilbury, L., Daridon, B., & Sukalac, K. (2019). General principles to justify plant biostimulant claims. Frontiers in Plant Science, 10, Article 494. https://doi.org/10.3389/fpls.2019.00494

Rouphael, Y., Cardarelli, M., Bonini, P., & Colla, G. (2017). Synergistic action of a microbial-based biostimulant and a plant derived-protein hydrolysate enhances lettuce tolerance to alkalinity and salinity. Frontiers in Plant Science, 8, Article 131. https://doi.org/10.3389/fpls.2017.00131

Shibaeva, T., Sherudilo, E., & Titov, A. (2018). Response of cucumber (Cucumis sativus L.) plants to prolonged permanent and short-term daily exposures to chilling temperature. Russian Journal of Plant Physiology, 65, 286–294. https://doi.org/10.1134/S1021443718020061

Teixeira, W. F., Fagan, E. B., Soares, L. H., Umburanas, R. C., Reichardt, K., & Neto, D. D. (2017). Foliar and seed application of amino acids affects the antioxidant metabolism of the soybean crop. Frontiers in Plant Science, 8, Article 327. https://doi.org/10.3389/fpls.2017.00327

Tkaczewska, J., Borawska-Dziadkiewicz, J., Kulawik, P., Duda, I., Morawska, M., & Mickowska, B. (2020). The effects of hydrolysis condition on the antioxidant activity of protein hydrolysate from Cyprinus carpio skin gelatin. LWT, 117, Article 108616. https://doi.org/10.1016/j.lwt.2019.108616

Xu, Y., Guo, S., Li, H., Sun, H., Lu, N., Shu, S., & Sun, J. (2017). Resistance of cucumber grafting rootstock pumpkin cultivars to chilling and salinity stresses. Horticultural Science and Technology, 35(2), 220–231. https://doi.org/10.12972/kjhst.20170025

Zhao, H., Zhang, K., Zhou, X., Xi, L., Wang, Y., Xu, H., Pan, T., & Zou, Z. (2017). Melatonin alleviates chilling stress in cucumber seedlings by up-regulation of CsZat12 and modulation of polyamine and abscisic acid metabolism. Scientific Reports, 7, Article 4998. https://doi.org/10.1038/s41598-017-05267-3

DOI: https://doi.org/10.5586/aa.756

Journal ISSN:
  • 2300-357X (online)
  • 0065-0951 (print; ceased since 2016)
This is an Open Access journal, which distributes its content under the terms of the Creative Commons Attribution License, which permits redistribution, commercial and non-commercial, provided that the content is properly cited.
The journal is a member of the Committee on Publication Ethics (COPE) and aims to follow the COPE’s principles.
The journal publisher is a member of the Open Access Scholarly Publishers Association.
The journal content is indexed in Similarity Check, the Crossref initiative to prevent scholarly and professional plagiarism.
Polish Botanical Society