Effect of Glycine Betaine on some Morphological Traits, Osmolyte Accumulations and Antioxidant System of Sports Grass under Salt Stress

Document Type : Research Article


Department of Horticultural Science, Faculty of Agriculture, Ilam University, Ilam, Iran


Grasses are narrow-leaved plants that are used as cover plants in landscape. These plants are one of the basic and necessary components of the green cover of most gardens, parks and as the background color of landscape. In Iran, due to the high costs of planting and management of grass, high water requirements, climatic incompatibility and damage to water and soil salinity, it is recommended to remove from the green space in some cities, especially in areas with low water and water and soil saline. If it is possible to benefit from the role and influence of these plants by observing the technical points and choosing the best species for each area. Salinity stress is the second limiting factor for the growth of plants in the world after drought, which affects the efficiency and performance of plants. Increase in salinity causes a decrease in the water potential in the soil. In this condition, the plant spends most of its energy to maintain the water potential, cell mass, and water absorption to have minimal growth. The aim of this research is the effect of external application of glycine betaine on the accumulation of osmolality compounds and the antioxidant system of sports grass under salt stress.
Materials and Methods
 This research was carried out in 2022 in pots in the research greenhouse of Ilam University as a factorial  based on a completely random design with three replications. Experimental treatments included three salinity levels with sodium chloride salt (without salinity, 50 and 100 mM sodium chloride) and three levels of glycine betaine foliar spraying (0, 5 and 10 mM). Glycine betaine application was performed after mowing twice with a distance of 48h from each other, and then salinity with sodium chloride salts was applied. 4 weeks after application of salinity stress, some morphological and biochemical characteristics of plants were  measured. The results were analysed using SAS software (v.9.2), and Tukey's test was used to compare the means at the 5% probability level.
Results and Discussion
The results showed that salinity stress decreased all the study morphological, physiological and biochemical parameters including plant height, shoot fresh and dry weight, number of tiller, leaf area, chlorophyll content, protein and total antioxidant capacity in the studied plants. It also increased peroxidase enzyme, H2O2 and proline in plants, but glycine betaine application significantly improved the morpho-physiological characteristics of plants compared to the control under salt stress conditions. Thus, the highest height, shoot fresh and dry weight, leaf area, number of tiller, chlorophyll content, and protein and antioxidant capacity were observed in plants sprayed with glycine betaine. Also, the highest content of glycine betaine and activity of catalase and peroxidase enzymes and the lowest content of glycine betaine and H2O2 were observed in in plants sprayed with glycine betaine and 10 mM glycine betaine was more effective than 5 mM. The occurrence of salinity in plants disrupts the absorption of ions and causes the reduction of nutrients and increases sodium ions. One of the effects of salinity in plants is the reduction of photosynthetic activity, which results in the reduction of chlorophyll, carbon dioxide absorption, photosynthetic capacity, plant height, shoot fresh and dry weight, number of tiller and leaf area. One of the most strategies to deal with stress is accumulation of osmolyte and increasing the antioxidant activity, which makes plants resistant to environmental stresses. Salinity, through the toxic effect of Na+ and Cl- ions, affects the growth and performance of the plant by reducing the soil water potential, disrupting water absorption and imbalance of nutrients in the plant. The results obtained from comparing the average results of glycine betaine show that glycine betaine increased plant height, shoot fresh and dry weight, number of tiller, leaf area, chlorophyll content, total protein and antioxidant capacity, but on the other hand, it increased proline and H2O2 decreased, which is due to the accumulation of glycine betaine as a protector in plants under salt stress conditions. In stress conditions, glycine betaine can protect photosynthetic activities including photosynthetic enzymes, proteins and lipids in thylakoid membranes in the combination of photosystem II, and also the task of protecting cell membranes against osmotic stresses in the plant.
The results obtained from this research showed that salinity stress reduced all the morphological, physiological and biochemical characteristics in the sport grass plants, but glycine betaine application played a positive role in reducing salinity damage and maintaining plant quality. Glycine betaine is known as one of the effective molecules in stress signaling, so it can protect the plant cells against stress by reducing the destruction of the membrane and by increasing the salt tolerance mechanisms. Also, glycine betaine 10 mM is introduced as the best treatment to reduce salinity damage in sport grass during present study.


Main Subjects

©2024 The author(s). This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source.

  1. Abid, M., Ali, S., Qi, L. K., Zahoor, R., Tian, Z., Jiang, D., & Dai, T. (2018). Physiological and biochemical changes during drought and recovery periods at tillering and jointing stages in wheat (Triticum aestivum). Scientific Reports, 8(1), 4615.‏ https://doi.org/10.1038/s41598-018-21441-7
  2. Alexieva, V., Sergiev, I., Mapelli, S., & Karanov, E. (2001). The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant, Cell & Environment24(12), 1337-1344.‏ https://doi.org/10.1046/j.1365-3040.2001.00778.x
  3. Ali, S., Abbas, Z., Seleiman, M.F., Rizwan, M., YavaŞ, İ., Alhammad, B.A., & Kalderis, D. (2020). Glycine betaine accumulation, significance and interests for heavy metal tolerance in plants. Plants9(7), 896.‏ https://doi.org/10.3390/plants9070896
  4. Amirjani, M.R. (2010). Effects of salinity stress on growth, mineral composition, proline content, antioxidant enzymes of soybean. American Journal of Physiology, 5(6), 350-360. https://doi.org/3923/ajpp.2010.350.360
  5. Apel, K., & Hirt, H. (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biology, 55, 373-399.‏ https://doi.org/1146/annurev.arplant.55.031903.141701
  6. Ashraf, M.F.M.R., & Foolad, M.R. (2007). Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environmental and Experimental Botany59(2), 206-216.‏ https://doi.org/10.1016/j.envexpbot.2005.12.006
  7. Ashraf, M.H.P.J.C., & Harris, P.J. (2013). Photosynthesis under stressful environments: An overview. Photosynthetica, 51, 163-190.‏ https://doi.org/10.1007/s11099-013-0021-6
  8. Bates, L.S., Waldren, R.A., & Teare, I.D. (1973). Rapid determination of free proline for water-stress studies. Plant and Soil39, 205-207. https://doi.org/10.1007/BF00018060
  9. Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry72(1-2), 248-254.‏ https://doi.org/10.1016/0003-2697(76)90527-3
  10. Cabrera, R.I. (2003). Demarcating salinity tolerance in greenhouse rose production. In International Symposium on Managing Greenhouse Crops in Saline Environment 609(pp. 51-57). (In Persian with English abstract). https://doi.org/10.17660/ActaHortic.2003.609.5
  11. Chen, T.H., & Murata, N. (2011). Glycinebetaine protects plants against abiotic stress: mechanisms and biotechnological applications. Plant, Cell & Environment, 34(1), 1-20.‏ https://doi.org/10.1111/j.1365-3040.2010.02232
  12. Craine, J.M. (2005). Reconciling plant strategy theories of Grime and Tilman. Journal of Ecology, 93(6), 1041-1052.‏ https://doi.org/10.1111/j.1365-2745.2005.01043.x
  13. Darvizheh, H., Zavareh, M., & Ghasemnezhad, M. (2017). Effects of prolin application on biochemistry characteristics of German chamomil (Matricaria chamomilla) in water stress. Applied Research of Plant Ecophysiology, 4(1), 35-60.‏ https://doi.org/10.22059/IJHS.2022.319917.1901
  14. De Oliveira, V.P., Marques, E.C., de Lacerda, C.F., Prisco, J.T., & Gomes Filho, E. (2013). Physiological and biochemical characteristics of Sorghum bicolor and Sorghum sudanense subjected to salt stress in two stages of development. African Journal of Agricultural Research, 8(8), 660-670.‏ https://doi.org/10.5897/AJAR12.861
  15. Doğan, M. (2011). Antioxidative & proline potentials as a protective mechanism in soybean plants under salinity stress. African Journal of Biotechnology10(32), 5972-5978.‏ https://doi.org/5897/AJB10.2114
  16. Dolatabadian, A., Sanavy, S.M., & Chashmi, N.A. (2008). The effects of foliar application of ascorbic acid (vitamin C) on antioxidant enzymes activities, lipid peroxidation and proline accumulation of canola (Brassica napus) under conditions of salt stress. Journal of Agronomy & Crop Science, 194(3), 206-213.‏ https://doi.org/10.1111/j.1439-037X.2008.00301.x
  17. Dudeck, A.E., Peacock, C.H., & Wildmon, J.C. (1993). Physiological and growth responses of St. Augustinegrass cultivars to salinity. HortScience Journal, 28(1), 46-48.‏ https://doi.org/10.21273/HORTSCI.28.1.46
  18. Eskandari, H., Ehsanpour, A.A., & Al Mansour, N. (2018). The effect of Rosmarinic acid on glycine betaine, carbohydrate and protein pattern changes of potato (Solanum tuberosum) callus under in vitro condition. Iranian Journal of Plant Biology, 10(2), 1-18.‏ (In Persian). https://doi.org/10.22108/IJPB.2017.107124.1058
  19. Farsaraei, S., Moghaddam, M., & Pirbalouti, A.G. (2020). Changes in growth and essential oil composition of sweet basil in response of salinity stress and superabsorbents application. Scientia Horticulturae271, 109465.‏ https://doi.org/10.1016/j.scienta.2020.109465
  20. Francois, L.E., Maas, E.V., Donovan, T.J., & Youngs, V.L. (1986). Effect of salinity on grain yield and quality, vegetative growth, and germination of semi‐dwarf and Durum wheat. Agronomy Journal, 78(6), 1053-1058.‏ https://doi.org/10.2134/agronj1986.00021962007800060023x
  21. Gan, L., Zhang, X., Liu, S., & Yin, S. (2018). Mitigating effect of glycinebetaine pretreatment on drought stress responses of creeping bentgrass. HortScience Journal, 53(12), 1842-1848.‏ https://doi.org/10.21273/HORTSCI13429-18
  22. Gapinska, M., Skodowska, M., & Gabara, B.(2008). Effect of short and long-term salinity on the activities of antioxidative enzymes and lipid peroxidation in tomato roots. Acta Physiologiae Plantarum, 30, 11-18. https://doi.org/10.1007/s11738-007-0072-z
  23. Giri, J. (2011). Glycinebetaine & abiotic stress tolerance in plants. Plant Signaling and Behavior6(11), 1746-1751.‏ https://doi.org/10.4161/psb.6.11.17801
  24. Haghighi, M., & Sheibanirad, A. (2018). Evaluating of Azealic Acid on Tomato Vegetative and Photosynthetic Parameters under Salinity Stress', Journal of Horticultural Science, 32(2), 287-300. https://org/10.22067/jhorts4.v32i2.64405
  25. Hatami, Z., Roein, Z., & Shiri, M.A. (2021). Alleviation of freezing injury to Coronilla varia ground cover by foliar application of glycine betaine. Journal of Plant Production, 28(4), 195-212.‏ (In Persian with English abstract). https://doi.org/10.22069/JOPP.2021.18569.2738
  26. Jamil, A., Riaz, S., Ashraf, M., & Foolad, M.R. (2011). Gene expression profiling of plants under salt stress. Critical Reviews in Plant Sciences30(5), 435-458. https://doi.org/10.1080/07352689.2011.605739
  27. Jing, Y.D., He, Z.L., & Yang, X.E. (2007). Role of soil rhizobacteria in phytoremediation of heavy metal contaminated soils. Journal of Zhejiang University Science B, 8, 192-207.‏ https://doi.org/10.1631/jzus.2007.B0192
  28. Joushan, Z., Sodeizadeh, H., Hakimzadeh Ardakani, M. A., Yazdani Biouki, R., & Khajahhosseini, S. (2020). Investigating the effect of foliar application of Glycine Betaine on some quantitative and qualitative characteristics of Mint (Mentha spicata( under salinity stress. Plant Productions, 43(2), 267-280.‏ (In Persian with English abstract). https://doi.org/10.22055/PPD.2019.27413.1666
  29. Kadkhodaie, A., Razmjoo, J., Zahedi, M., & Pessarakli, M. (2014). Selecting sesame genotypes for drought tolerance based on some physiochemical traits. Agronomy Journal106(1), 111-118.‏ https://doi.org/10.2134/agronj2013.0260
  30. Katerji, N., Van Hoorn, J.W., Hamdy, A., Karam, F., & Mastrorilli, M. (1996). Effect of salinity on water stress, growth, and yield of maize and sunflower. Agricultural Water Management30(3), 237-249.‏ https://doi.org/10.1016/0378-3774(95)01228-1
  31. Khandan-Mirkohi, A., Baie, N., & Hadavi, E. (2016). The effect of reduced application of water and nitrogen on growth management of mixed Turf-Grass. Journal of Horticultural Science, 30(2), 327-335. (In Persian). https://doi.org/10.22067/jhorts4.v30i2.43607
  32. Leopold, A.C. (1984). Evidence for toxicity effects of salt on membranes. Salinity tolerance in plants. Strategies for Crop Improvement, 67-76. https://doi.org/10.1626/jcs.64.93
  33. Leopold, A.C., Sun, W.Q., & Bernal-Lugo, I. (1994). The glassy state in seeds: analysis and function. Seed Science Research, 4(3), 267-274. https://doi.org/22055/PPD.2019.27413.1666
  34. Lichtenthaler, H.K. (1987). Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. In Methods in enzymology, 148, 350-382). Academic Press. https://doi.org/10.1016/0076-6879(87)48036-1
  35. Maghsoumi Holasoo, S., & Pourakbar, L. (2014). The effects of salinity stress on the growth and some physiological parameters of wheat (Triticum aestivum) seedlings. Iranian Journal of Plant Biology, 6(19), 31-42.‏ (In Persian). https://doi.org/20.1001.1.20088264.1393.
  36. Miri, H.R., & Zamani Moghadam, A. (2014). The effect of external usage of glycine betaine on corn (Zea mays) in drought condition. Iranian Journal of Field Crops Research12(4), 704-717.‏https://doi.org/10.22067/GSC.V12I4.24221
  37. Mohseni Mohammadjanlou, A., Seyed Sharifi, R., & Khomari, S. (2023). Effects of bio-fertilizer and putrescine on yield and some agrophysiological and biochemical traits of wheat under soil salinity stress. Environmental Stresses in Crop Sciences.‏ (In Persian). https://doi.org/10.22077/escs.2023.4774.2067
  38. Mohammadi, M., Aelaei, M., Saidi, M., (2021). Pre‑harvest spray of GABA and spermine delays postharvest senescence and alleviates chilling injury of gerbera cut flowers during cold storage. Scientific Reports, 11, 14166. https://doi.org/10.1038/s41598-021-93377-4
  39. Mohammadi, M, Eghlima Gh, Ranjbar ME (2023). Ascorbic acid reduces chilling injury in anthurium cut flowers during cold storage by increasing salicylic acid biosynthesis. Postharvest Biology Technology, 201, 112359. https://doi.org/10.1016/j.postharvbio.2023
  40. Moller, I.M., Jensen, P.E., & Hansson, A. (2007). Oxidative modifications to cellular components in plants. Annu. Rev. Plant Biology58, 459-481. https://doi.org/10.1146/annurev.arplant.58.032806.103946
  41. ‏Mortezainajad, F., Khavarinajad, R.A., & Emami, M. (2005). Evaluation of some performance parameters and proline rice varieties under salt stress. New Agricultural Science2(4), 65-70. (In Persian). https://doi.org/10.22055/PPD.2019.27413.1666
  42. Nawaz, M., & Wang, Z. (2020). Abscisic acid and glycine betaine mediated tolerance mechanisms under drought stress and recovery in Axonopus compressus: a new insight. Scientific Reports, 10(1), 6942.‏ https://doi.org/10.1038/s41598-020-63447-0
  43. Papageorgiou, G.C., & Murata, N. (1995). The unusually strong stabilizing effects of glycine betaine on the structure and function of the oxygen-evolving photosystem II complex. Photosynthesis Research44, 243-252.‏ https://doi.org/10.1007/BF00048597
  44. Papageorgiou, G.C., Fujimura, Y., & Murata, N. (1991). Protection of the oxygen-evolving photosystem II complex by glycinebetaine. Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1057(3), 361-366.‏ https://doi.org/10.1016/S0005-2728(05)80148-3
  45. Park, E.J., Jeknic, Z., & Chen, T.H. (2006). Exogenous application of glycinebetaine increases chilling tolerance in tomato plants. Plant and Cell Physiology47(6), 706-714.‏ https://doi.org/10.1093/pcp/pcj041
  46. Promyou, S., Ketsa, S., & van Doorn, W.G.(2012). Salicylic acid alleviates chilling injury in anthurium (Anthurium andraeanum) flowers. Postharvest Biology Technology, 64, 104–110. https://doi.org/10.1016/j.postharvbio.2011.10.002
  47. Sairam, R.K., & Srivastava, G.C.(2002). Changes in antioxidant activity in sub-cellular fraction of tolerant and susceptible wheat genotypes in response to long term salt stress. Plant Science, 162, 897-904. https://doi.org/1016/S0168-9452(02)00037-7
  48. Shahzad, K., Hussain, S., Arfan, M., Hussain, S., Waraich, E.A., Zamir, S., & El-Esawi, M.A. (2021). Exogenously applied gibberellic acid enhances growth and salinity stress tolerance of maize through modulating the morpho-physiological, biochemical and molecular attributes. Biomolecules11(7), 1005.‏  https://doi.org/10.3390/biom11071005
  49. Yazdani-Biouki, R., & Beyrami, H. (2020). Investigating the effects of Glycine betaine on growth and flower yield of Damask Rose under salinity stress. Journal of Crops Improvement, 22(1), 119-134. (In Persian with English abstract). https://doi.org/10.22059/jci.2020.287265.2257
  50. Yunwei, D., Tingting, J., & Shuanglin, D. (2007). Stress responses to rapid temperature changes of the juvenile sea cucumber (Apostichopus japonicus Selenka). Journal of Ocean University of China, 6(3), 275-280.‏ https://doi.org/10.1007/s11802-007-0275-3
  51. Zadehbagheri, M., & Salehi Salmi, M.R. (2016). The physiological, morphological and bio-chemical comparison of the current grass Shiraz city’s green space with tall Fescue (Festuca arundinacea Schreb). Isfahan University of Technology-Journal of Crop Production and Processing5(18), 15-25.‏ (In Persian). https://doi.org/10.18869/acadpub.jcpp.5.18.15
  52. Zamani, M.M., Rabiyei, V., & Nejatian, M.A. (2013). Effect of proline and glycine betaine application on some physiological characteristics in grapevine under drought stress. Iranian Journal of Horticultural Science43(4).‏ (In Persian with English abstract). https://doi.org/22059/IJHS.2012.29374  
  53. Zhang, Z., Huber, D.J., & Rao, J. (2013). Antioxidant systems of ripening avocado (Persea americana) fruit following treatment at the preclimacteric stage with aqueous 1-methylcyclopropene. Postharvest Biology and Technology76, 58-64.‏ https://doi.org/10.1016/j.postharvbio.2012.09.003
  • Receive Date: 28 November 2023
  • Revise Date: 03 January 2024
  • Accept Date: 09 January 2024
  • First Publish Date: 09 January 2024