The effect of biological treatments of putrescine and salicylic acid on enhancing rosemary resistance to salinity through activation of secondary metabolites

Document Type : Research Paper

Authors

1 MSc, Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, Iran

2 Corresponding Author, Prof., Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, Iran, E-mail: samsampoor@hormozgan.ac.ir

3 Assistant Prof., Department of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, Iran

10.22092/ijfrpr.2025.370001.1678

Abstract

Background and Objective: Soil salinity is one of the most important abiotic stresses that restricts plant growth and yield by disturbing ionic balance, reducing water absorption, and increasing the production of reactive oxygen species (ROS). Rosemary (Rosmarinus officinalis L.), as a medicinal and aromatic plant, has a high capacity for producing secondary metabolites such as phenols, flavonoids, and essential oils, which play an essential role in plant defense against stresses. Recent studies have shown that the application of growth-regulating compounds such as putrescine (a natural polyamine) and salicylic acid (a key phytohormone) can enhance plant resistance to unfavorable conditions by stimulating defense pathways and increasing the production of antioxidant compounds. Therefore, the aim of this research was to evaluate the effect of biological treatments with putrescine and salicylic acid on rosemary tolerance to salinity through the induction of secondary metabolites and the improvement of defense mechanisms.
Methodology: In this study, the effects of putrescine (at concentrations of 0, 0.5, 1, and 2 mM) and salicylic acid (at concentrations of 0, 0.5, 0.7, and 1 mM) under salinity stress (0, 75, 125, and 150 mM NaCl) were evaluated on the physiological traits, biochemical parameters, and metabolite variations of rosemary. The experiment was conducted as a factorial in a completely randomized design with three replications.
Results: The results showed that with increasing salinity concentration, proline content in the plant increased significantly. Under salinity stresses of 75, 125, and 150 mM, treatments with putrescine at 2 mM and salicylic acid at 1 mM increased proline content by 0.138, 0.166, and 0.218 mg g⁻¹ fresh weight, respectively, corresponding to increases of 78.98%, 82.53%, and 59.91% compared to the control. In addition, salinity increment caused a significant increase in carbohydrate content, and putrescine treatment at 0.5 mM under non-stress conditions enhanced carbohydrate accumulation. Under 75 and 125 mM salinity, the highest carbohydrate content was observed in the salicylic acid treatment at 1 mM, reaching 3.13 and 3.38 mg g⁻¹ fresh weight, which corresponded to increases of 138.93% and 77.89% compared to the control. Under 150 mM salinity, the combined treatment of putrescine (0.5 mM) and salicylic acid (0.7 mM) resulted in the highest proline content (3.3 mg g⁻¹). Under non-stress conditions, putrescine at 0.5 mM increased peroxidase activity by 86.53%. At 75 mM salinity, putrescine at 1 mM enhanced peroxidase activity by 23.47%, whereas at 150 mM salinity, the combined treatment of putrescine (0.5 mM) and salicylic acid (0.7 mM) increased peroxidase activity by 116.1%. Furthermore, the analysis of defense-related compounds showed that the combined treatment of putrescine (2 mM) and salicylic acid (1 mM) resulted in the highest rosmarinic acid content under 150 mM salinity. This treatment also significantly increased terpenoid compounds such as α-pinene, 1,8-cineole, borneol, and camphor. The increase in 1,8-cineole under this treatment indicates its positive effect on the synthesis of volatile compounds with antioxidant and antimicrobial properties.
Conclusion: The findings of this study demonstrated that biological treatments with putrescine and salicylic acid play a significant role in enhancing the salinity tolerance of rosemary plants. These compounds improved the plant’s physiological balance under salt stress by increasing the production of secondary metabolites such as phenols and flavonoids, reducing oxidative damage, and activating antioxidant-related defense pathways. Therefore, the application of these treatments can be recommended as an eco-friendly, low-cost, and effective strategy to improve the tolerance of medicinal and aromatic plants to environmental stresses, especially salinity, within sustainable agriculture programs and the production of high-value medicinal crops.

Keywords

References

-Ahmad, R., Hussain, S., Anjum, M.A., Khalid, M.F., Saqib, M., Zakir, I., Hassan, A., Fahad, S. and Ahmad, S., 2019. Oxidative stress and antioxidant defense mechanisms in plants under salt stress. Plant abiotic stress tolerance: Agronomic, molecular and biotechnological approaches, pp.191-205. https://doi.org/10.1007/978-3-030-06118-0_8
-Ahmed, M., Tóth, Z. and Decsi, K., 2024. The impact of salinity on crop yields and the confrontational behavior of transcriptional regulators, nanoparticles, and antioxidant defensive mechanisms under stressful conditions: A review. International Journal of Molecular Sciences, 25(5): 2654. https://doi.org/10.3390/ijms25052654
-Al-Fraihat, A.H., Al-Dalain, S.Y., Zatimeh, A.A. and Haddad, M.A., 2023. Enhancing rosemary (Rosmarinus officinalis L.) growth and volatile oil constituents grown under soil salinity stress by some amino acids. Horticulturae, 9(2): 252. https://doi.org/10.3390/horticulturae9020252
-Ali, A., Kant, K., Kaur, N., Gupta, S., Jindal, P., Gill, S.S. and Naeem, M., 2024. Salicylic acid: Homeostasis, signalling and phytohormone crosstalk in plants under environmental challenges. South African Journal of Botany, 169: 314-335. https://doi.org/10.1016/j.sajb.2024.04.012
-Arif, Y., Singh, P., Siddiqui, H., Bajguz, A. and Hayat, S., 2020. Salinity induced physiological and biochemical changes in plants: An omic approach towards salt stress tolerance. Plant Physiology and Biochemistry, 156: 64-77. https://doi.org/10.1016/j.plaphy.2020.08.042
-Bates, L.S., Waldren, R.P.A. and Teare, I.D., 1973. Rapid determination of free proline for water-stress studies. Plant and soil, 39: 205-207. http://dx.doi.org/10.1007/BF00018060
-Boorboori, M.R. and Li, J., 2025. The effect of salinity stress on tomato defense mechanisms and exogenous application of salicylic acid, abscisic acid, and melatonin to reduce salinity stress. Soil Science and Plant Nutrition, 71(1): 93-110. https://doi.org/10.1080/00380768.2024.2405834
-Bulgari, R., Morgutti, S., Cocetta, G., Negrini, N., Farris, S., Calcante, A., Spinardi, A., Ferrari, E., Mignani, I., Oberti, R. and Ferrante, A., 2017. Evaluation of borage extracts as potential biostimulant using a phenomic, agronomic, physiological, and biochemical approach. Frontiers in Plant Science, 8: 935.
-Chance, B. and Maehly, A. C., 1955. Assay of catalases and peroxidases, 2: 764-775. https://doi.org/10.1002/9780470110171.ch14
-Ehtaiwesh, A.F., 2022. The effect of salinity on nutrient availability and uptake in crop plants. Scientific Journal of Applied Sciences of Sabratha University, 9(9): 55-73. https://doi.org/10.47891/sabujas.v0i0.55-73
-Farzane, A., Nemati, H., Shoor, M. and Ansari, H., 2020. Antioxidant enzyme and plant productivity changes in field-grown tomato under drought stress conditions using exogenous putrescine. Journal of Plant Physiology and Breeding, 10(1): 29-40.
-Garoosi, M.K., Sanjarian, F. and Chaichi, M., 2023. The role of γ-aminobutyric acid and salicylic acid in heat stress tolerance under salinity conditions in Origanum vulgare L. Plos one, 18(7), p.e0288169.
-Golkar, P., Taghizadeh, M. and Yousefian, Z., 2019. The effects of chitosan and salicylic acid on elicitation of secondary metabolites and antioxidant activity of safflower under in vitro salinity stress. Plant Cell, Tissue and Organ Culture (PCTOC), 137(3): 575-585. https://doi.org/10.1007/s11240-019-01592-9
-Hosseini, T., Shekari, F. and Ghorbanli, M., 2010. Effect of salt stress on ion content, proline and antioxidative enzymes of two safflower cultivars (Carthamus tinctorius L.). J. Food Agric. Environ, 8(2): 1080-1086.
-Hussein, O.S. and Abdelkadr, A., 2024. Metabolomics Profiling of Chilled (Coriandrum sativum L.) Primed by Silicate, Humic acid and Gamma Radiation. Arab Journal of Nuclear Sciences and Applications, 57(2): 85-99. https://doi.org/10.21608/ajnsa.2024.250145.1793
-Isah, T., 2019. Stress and defense responses in plant secondary metabolites production. Biological research, 52(39): 1-25. https://doi.org/10.1186/s40659-019-0246-3
-Jiménez, A.D.C., 2019. Biological, Chemical, and Physical Investigation of Natural Terpenes (Doctoral dissertation, Texas Southern University).
-Li Pomi, F., Papa, V., Borgia, F., Vaccaro, M., Allegra, A., Cicero, N. and Gangemi, S., 2023. Rosmarinus officinalis and skin: antioxidant activity and possible therapeutical role in cutaneous diseases. Antioxidants, 12(3): 680. https://doi.org/10.3390/antiox12030680
-Mahajan, M. and Pal, P.K., 2023. Drought and salinity stress in medicinal and aromatic plants: Physiological response, adaptive mechanism, management/amelioration strategies, and an opportunity for production of bioactive compounds. Advances in Agronomy, 182: 221-273. https://doi.org/10.1016/bs.agron.2023.06.005
-Majeed, A. and Muhammad, Z., 2019. Salinity: a major agricultural problem—causes, impacts on crop productivity and management strategies. Plant abiotic stress tolerance: Agronomic, molecular and biotechnological approaches, pp.83-99. https://doi.org/10.1007/978-3-030-06118-0_3
-Mohammadi, M., Nezamdoost, D., Khosravi Far, F., Zulfiqar, F., Eghlima, G. and Aghamir, F., 2024. Exogenous putrescine application imparts salt stress-induced oxidative stress tolerance via regulating antioxidant activity, potassium uptake, and abscisic acid to gibberellin ratio in Zinnia flowers. BMC Plant Biology, 24(1): 865. https://doi.org/10.1186/s12870-024-05560-0
-Mohammed, H.A., Emwas, A.H. and Khan, R.A., 2023. Salt-tolerant plants, halophytes, as renewable natural resources for cancer prevention and treatment: roles of phenolics and flavonoids in immunomodulation and suppression of oxidative stress towards cancer management. International Journal of Molecular Sciences, 24(6): 5171. https://doi.org/10.3390/ijms24065171
-Misra, N. and Saxena, P., 2009. Effect of salicylic acid on proline metabolism in lentil grown under salinity stress. Plant Science, 177(3): 181-189. https://doi.org/10.1016/j.plantsci.2009.05.007
-Mutlu, S.A.L.I.H., Atici, Ö. and Nalbantoglu, B., 2009. Effects of salicylic acid and salinity on apoplastic antioxidant enzymes in two wheat cultivars differing in salt tolerance. Biologia Plantarum, 53: 334-338. https://doi.org/10.1007/s10535-009-0061-8
-Nahar, K., Hasanuzzaman, M. and Fujita, M., 2016. Roles of osmolytes in plant adaptation to drought and salinity. Osmolytes and plants acclimation to changing environment: Emerging omics technologies, pp.37-68. https://doi.org/10.1007/978-81-322-2616-1_4
-Noreen, S., Ashraf, M., Hussain, M. and Jamil, A., 2009. Exogenous application of salicylic acid enhances antioxidative capacity in salt stressed sunflower (Helianthus annuus L.) Pakistan Journal of Botany, 41(1): 473-479.
-Sadia, H., Shahbaz, M., Kiran, A. and Saleem, M.F., 2023. Interactive effect of salicylic acid and ascorbic acid on gaseous exchange and mineral nutrients of chicory (Cichorium intybus L.) under saline conditions. Pak J Bot, 55(6): 1999-2012. https://doi.org/10.30848/PJB2023-6(22)
-Salam, U., Ullah, S., Tang, Z.H., Elateeq, A.A., Khan, Y., Khan, J., Khan, A. and Ali, S., 2023. Plant metabolomics: an overview of the role of primary and secondary metabolites against different environmental stress factors. Life, 13(3): 706. https://doi.org/10.3390/life13030706
-Sattarzadeh, E., Yarnia, M., Khalilvand Behrooznia, E., Mirshekari, B. and Rashidi, V., 2023. Investigation of the possibility of reducing the effects of low irrigation of lavender (Lavandula officinalis L.) using biofertilizers and phosphorus through changes in some morphological and biochemical characteristics. Environmental Stresses in Crop Sciences, 16(4): 1153-1171.
-Schubert, S. and Qadir, M., 2024. Soil Salinity and Salt Resistance of Crop Plants. Springer International Publishing. https://doi.org/10.1007/978-3-031-73250-8
-Shankar, A., Ali, A., Abdullah, H.M., Balaji, J., Kaur, J., Saeed, F., Wasiq, M., Imran, A., Jibraeel, H., Raheem, M.S. and Aslam, A., 2024. Nutritional Composition, Phytochemical Profile, Therapeutic Potentials, and Food Applications of Rosemary: A Comprehensive Review. Journal of Food Composition and Analysis, 135: 1-12.
-Silva, S., Costa, E. M., Calhau, C., Morais, R. M. and Pintado, M.E., 2017. Anthocyanin extraction from plant tissues: A review. Critical reviews in food science and nutrition, 57(14): 3072-3083. https://doi.org/10.1080/10408398.2015.1087963
-Turkyilmaz Unal, B., Mentis, O. and Akyol, E., 2015. Effects of exogenous salicylic acid on antioxidant activity and proline accumulation in apple (Malus domestica L.). Horticulture, Environment, and Biotechnology, 56: 606-611. https://doi.org/10.1007/s13580-015-0049-6
-Yemm, E.W. and Willis, A., 1954. The estimation of carbohydrates in plant extracts by anthrone. Biochemical journal, 57(3): 508-514. https://doi.org/10.1042/bj0570508
-Zhong, M., Yue, L., Liu, W., Qin, H., Lei, B., Huang, R., Yang, X. and Kang, Y., 2023. Genome-wide identification and characterization of the polyamine uptake transporter (Put) gene family in tomatoes and the role of Put2 in response to salt stress. Antioxidants, 12(2): 1-24. https://doi.org/10.3390/antiox12020228
-Zulfiqar, F. and Ashraf, M., 2023. Proline alleviates abiotic stress induced oxidative stress in plants. Journal of Plant Growth Regulation, 42(8): 4629-4651. https://doi.org/10.1007/s00344-022-10839-3

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History

  • Receive Date: 06 July 2025
  • Revise Date: 28 August 2025
  • Accept Date: 14 September 2025

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