Glycine-betaine induced salinity tolerance in maize by regulating the physiological attributes, antioxidant defense system and ionic homeostasis

  • Zain DUSTGEER University of Agriculture Faisalabad, Department of Botany, Faisalabad, 38040
  • Mahmoud F. SELEIMAN King Saud University, College of Food and Agriculture Sciences, Plant Production Department, P.O. Box 2460, Riyadh 11451; Menoufia University, Faculty of Agriculture, Department of Crop Sciences, Shibin El-kom 32514
  • Imran KHAN University of Agriculture Faisalabad, Department of Agronomy, Faisalabad, 38040
  • Muhammad U. CHATTHA University of Agriculture Faisalabad, Department of Agronomy, Faisalabad, 38040
  • Esmat F. ALI Taif University, College of Science, Department of Biology, P.O. Box 11099, Taif 21944
  • Bushra A. ALHAMMAD Prince Sattam Bin Abdulaziz University, College of Science and Humanity Studies, Biology Department, Al Kharj Box 292, Riyadh 11942
  • Rewaa S. JALAL University of Jeddah, College of Sciences, Department of Biology, Jeddah
  • Yahya REFAY King Saud University, College of Food and Agriculture Sciences, Plant Production Department, P.O. Box 2460, Riyadh 11451
  • Muhammad U. HASSAN University of Agriculture Faisalabad, Department of Agronomy, Faisalabad, 38040
Keywords: antioxidants; growth, glycine-betaine; nutrients accumulation; photosynthesis; salt stress; soluble proteins


The plants are exposed to different abiotic stresses, including the salinity stress (SS) that negatively affect the growth, metabolism, physiological and biochemical processes. Thus, this study investigated the effect of diverse levels of foliar-applied GB (0 control, 50 mM and 100 mM) on maize growth, membrane stability, physiological and biochemical attributes, antioxidant enzymes and nutrients accumulation under different levels of SS (i.e., control, 6 dS m-1, 12 dS m-1). Salt stress diminished the root and shoot length, root and shoot biomass, chlorophyll contents, photosynthetic rate (Pn), stomatal conductance (gs), relative water contents (RWC), soluble proteins (SP) and free amino acids; (FAA); and increased activities of antioxidant enzymes, electrical conductivity (EC) and accumulation of malondialdehyde (MDA), hydrogen peroxide (H2O2), Na+ and Cl ions. GB application significantly increased root and shoot growth, leaves per plant, shoots length, chlorophyll contents, gs, Pn and membrane stability by reducing MDA and H2O2 accumulation. Moreover, GB also increased the SP, FAA accumulation, activities of antioxidant enzymes and Na+ and Cl- exclusion by favouring Ca2+ and K+ accumulation. In conclusion, the foliar-applied GB increased Pn, gs, ant-oxidants activities, and accumulation of SP and FAA; and reduced the accretion of Na+ and Cl by favouring the Ca2+ and K+ accretion which in turns improved growth under SS.


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Aebi H (1984). Catalase in vitro. Methods in Enzymology 105:121-126.

Abbas W, Ashraf M, Akram NA (2010). Alleviation of salt-induced adverse effects in eggplant (Solanum melongena L.) by glycinebetaine and sugarbeet extracts. Scientia Horticulturae 125(3):188-195.

Abd-El-Mageed TA, Semida WM, Rady MM (2017). Moringa leaf extract as biostimulant improves water use efficiency, physio-biochemical attributes of squash plants under deficit irrigation. Agriculture and Water Management 193:46-54.

Ahmed N, Zhang Y, Li K, Zhou Y, Zhang M, Li M (2019). Exogenous application of glycine betaine improved water use efficiency in winter wheat (Triticum aestivum L.) via modulating photosynthetic efficiency and antioxidative capacity under conventional and limited irrigation conditions. Crop Journal 7(5):635-650.

Alasvandyari F, Mahdavi B, Hosseini S (2017). Glycine betaine affects the antioxidant system and ion accumulation and reduces salinity-induced damage in safflower seedlings. Archives of Biological Sciences 69:139-147.

Al-Ashkar, I, Alderfasi A, El-Hendawy S, Al-Suhaibani N, El-Kafafi S, Seleiman MF (2019). Detecting salt tolerance in doubled haploid wheat lines. Agronomy 9(4):211.

Al-Ashkar I, Alderfasi A, Romdhane WB, Seleiman MF, El-Said RA, Al-Doss A (2020). Morphological and genetic diversity within salt tolerance detection in eighteen wheat genotypes. Plants 9(3):287.

Ali S, Abbas ZM, Seleiman MF, Rizwan M, Yavas L, Alhammad BA, … Kalderis D (2020). Glycine betaine accumulation, significance and interests for heavy metal tolerance in plants. Plants 7:896.

Aamer M, Muhammad UH, Li Z, Abid A, Su Q, Liu Y, … Huang G (2018). Foliar application of glycinebetaine (gb) alleviates the cadmium (cd) toxicity in spinach through reducing cd uptake and improving the activity of anti-oxidant system. Applied Ecology and Environmental Research 1:7575-83.

Apel K, Hirt H (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annual Review of Plant Biology 55:373-399. https://doi.rog/10.1146/annurev.arplant.55.031903.141701

Ashraf M (2004). Some important physiological selection criteria for salt tolerance in plants. Functional Ecology of Plants 199(5):361-376.

Blum A (2017) Osmotic adjustment is a prime drought stress adaptive engine in support of plant production. Plant Cell and Environment 40(1):4-10.

Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72(1-2):248-254.

Caparrós P, Llanderal A, Hegarat E, Jiménez-Lao M, Lao MT (2020). Effects of exogenous application of osmotic adjustment substances on growth, pigment concentration, and physiological parameters of Dracaena sanderiana under different levels of salinity. Agronomy 10:125.

Cirillo C, Rouphael Y, Caputo R, Raimondi G, Sifola MI, Pascale SD (2016). Effects of high salinity and the exogenous application of an osmolyte on growth, photosynthesis, and mineral composition in two ornamental shrubs. Journal of Horticulture Science and Biotechnology 91:14-22.

Demiral T, Turkan P (2004). Does exogenous glycinebetaine affect antioxidative system of rice seedlings under NaCl treatment? Journal of Plant Physiology 161(10):1089-1100.

Gadallah MAA (1999). Effects of proline and glycinebetaine on Vicia faba responses to salt stress. Biologia Plantarum 42(2):249-257.

Habib N, Ashraf M, Ali Q, Perveen R (2012). Response of salt stressed okra (Abelmoschus esculentus Moench) plants to foliar-applied glycine betaine and glycine betaine containing sugar beet extract. South African Journal of Botany 83:151-158.

Hamilton PB, Van-Slyke DD (1943). Amino acid determination with ninhydrin. Journal of Biological Chemistry 150(1):231-250.

Hasanuzzaman M, Alam MM, Rahman A, Hasanuzzaman M, Nahar K, Fujita M (2014). Exogenous proline and glycine betaine mediated upregulation of antioxidant defense and glyoxalase systems provides better protection against salt-induced oxidative stress in two rice (Oryza sativa L.) varieties. Biomed Research International 1:1-15.

Homer DC, Pratt PF (1961). Methods of Analysis for soils, plants and waters. University of California, Davis.

Hoque MA, Banu MNA, Okuma E, Amako K, Nakamura Y, Shimoishi Y, Murata Y (2007). Exogenous proline and glycinebetaine increase NaCl-induced ascorbateglutathione cycle enzyme activities, and proline improves salt tolerance more than glycinebetaine in tobacco Bright Yellow-2 suspension-cultured cells. Journal of Plant Physiology 164(11):1457-1468.

Hu L, Hu T, Zhang X, Pang X, Fu J (2012). Exogenous glycine betaine ameliorates the adverse effect of salt stress on perennial ryegrass. Journal of the American Society of Horticulture Science 137(1):38-46.

Jamil A, Riaz S, Ashraf M, Foolad MR (2011). Gene expression profiling of plants under salt stress. Critical Review in Plant Sciences 30(5):435-458.

Jones J B, Case VW (1990). Sampling, handling, and analyzing plant tissue samples. Soil Test and Plant Analysis 3:389-427.

Lee DH, Kim YS, Lee CB (2001). The inductive responses of the antioxidant enzymes by salt stress in the rice (Oryza sativa L.). Journal of Plant Physiology 158(6):737-745.

Livia SS, Gabor K, Soltan S (2002). Effect of salt stress on free amino acid and polyamine content in cereal. Acta Biologica Szegediensis 46(3-4):73-75.

Lichtenthaler HK (1987). Chlorophylls and carotenoids: Pigments of photosynthetic biomembrane. Methods in Enzymology 148:350-352.

Kaya C, O Sönmez, S Aydemir M Dikilitaş (2013). Mitigation effects of glycinebetaine on oxidative stress and some key growth parameters of maize exposed to salt stress. Turkish Journal Agriculture and Forestry 37(2):188-194.

Mahmood T, Ashraf M, Shahbaz M (2009) Does exogenous application of glycinebetaine as a pre-sowing seed treatment improve growth and regulate some key physiological attributes in wheat plants grown under water deficit conditions? Pakistan Journal of Botany 41(3):1291-1302.

Ma QQ, Wang W, Li YH, Li DQ, Zou Q (2006). Alleviation of photoinhibition in drought-stressed wheat (Triticum aestivum) by foliar-applied glycinebetaine. Journal of Plant Physiology 163(2):165-175.

Mansour MMF (1998). Protection of plasma membrane of onion epidermal cells by glycinebetaine and proline against NaCl stress. Plant Physiology and Biochemistry 36(10):767-772.

Maxwell K, Johnson GN (2000). Chlorophyll fluorescence: a practical guide. Journal of Experimental Botany 51(345):659-668.

Mbarki S, Sytar O, Cerda A, Zivcak M, Rastogi A, He X, … Brestic C (2018). Strategies to mitigate the salt stress effects on photosynthetic apparatus and productivity of crop plants. In: Salinity Responses and Tolerance in Plants 1:85-136.

Meloni DA, Martínez CA (2009). Glycinebetaine improves salt tolerance in vinal (Prosopis ruscifolia Griesbach) seedlings. Brazilian Journal of Plant Physiology 21(3):233-241.

Mittler R (2002). Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science 7(9):405-410.

Mostofa MG, Fujita M (2013). Salicylic acid alleviates copper toxicity in rice seedlings by up-regulating antioxidative and glyoxalase systems. Ecotoxicology 22(6):959-973.

Moustakas M, Sperdouli I, Kouna T, Antonopoulou C, Therios I (2011). Exogenous proline induces soluble sugar accumulation and alleviates drought stress effects on photosystem II functioning of Arabidopsis thaliana leaves. Plant Growth Regulation 65(2):315-321.

Mukherjee SP, Choudhuri MA (1983). Implications of water stress induced changes in the levels of endogenous ascorbic acid and hydrogen peroxide in Vigna seedlings. Physiology of Plants 58:166-170.

Munns R, Tester M (2008). Mechanisms of salinity tolerance. Annual Review of Plant Biology 59:651-681.

Nawaz K, Ashraf M (2007). Improvement in salt tolerance of maize by exogenous application of glycinebetaine: growth and water relations. Pakistan Journal of Botany 39(5):1647-1653.

Radi R (2018). Oxygen radicals, nitric oxide, and peroxynitrite: Redox pathways in molecular medicine. Proceedings of the National Academy of Science, USA 115:5839-5848.

Rady MO, Semida VM, Abd-El-Mageed T, Hemida KA, Rady MM (2018). Up-regulation of antioxidative defense systems by glycine betaine foliar application in onion plants confer tolerance to salinity stress. Scientia Horticulturae 240:614-622.

Rahman MS, Miyake H, Takeoka Y (2002). Effects of exogenous glycinebetaine on growth and ultrastructure of salt-stressed rice seedlings (Oryza sativa L.). Plant Production Science 5(1):33-44.

Ranganayakulu GS, Veeranagamallaiah G, Sudhakar C (2013). Effect of salt stress on osmolyte accumulation in two groundnut cultivars (Arachis hypogaea L.) with contrasting salt tolerance. African Journal of Plant Science 7(12):586-592.

Rao KM, Sresty TVS (2000). Antioxidative parameters in the seedlings of pigeonpea (Cajanus cajan L.) Millspaugh) in response to Zn and Ni stresses. Plant Science 157(1):113-128.

Raza SH, Athar HR, Ashraf M, Hameed A (2007). Glycinebetaine-induced modulation of antioxidant enzymes activities and ion accumulation in two wheat cultivars differing in salt tolerance. Environmental and Experimental Botany 60(3):368-376.

Sakamoto A, Murata N (2002). The role of glycine betaine in the protection of plants from stress: clues from transgenic plants. Plant Cell and Environment 25(2):163-171.

Sakr MT, El-Sarkassy NM, PFuller M (2012). Osmoregulators proline and glycine betaine counteract salinity stress in canola. Agronomy for Sustainable Development 32(3):747-754.

Semida WM, Abd El-mageed TA, Howladar SM, Rady MM (2016). Foliar-applied ɑ-tocopherol enhances salt-tolerance in onion plants by improving antioxidant defense system. Australian Journal of Crop Sciences 10(7):1030-1039.

Semida WM, El-Mageed TAA, Mohamed SE, El-Sawah NA (201)7. Combined effect of deficit irrigation and foliar-applied salicylic acid on physiological responses, yield, and water-use efficiency of onion plants in saline calcareous soil. Archives of Agronomy and Soil Science 63(9):1227-1239.

Semida WM, Taha RS, Abdelhamid MT, Rady MM (2014). Foliar-applied α-tocopherol enhances salt-tolerance in Vicia faba L. plants grown under saline conditions. South African Journal of Botany 95:24-31.

Shrivastava, P, Kumar R. 2015. Soil salinity: a serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi Journal of Biological Sciences 22(2):123-131.

Seleiman MF, Al-Shuaibani N, Ali N, Akmal M, Alotaibi M, Rafey Y, … Battaglia ML (2021). Drought stress impacts on plants and different approaches to alleviate its adverse effects. Plants 10(2):259.

Seleiman MF, Kheir AMS (2018). Saline soil properties, quality and productivity of wheat grown with bagasse ash and thiourea in different climatic zones. Chemosphere 193:538-546.

Seleiman MF, Santanen A, Jaakkola S, Ekholm P, Hartikainen H, Stoddard FL, Mäkelä PS (2013). Biomass yield and quality of bioenergy crops grown with synthetic and organic fertilizers. Biomass Bioenergy 59:477-485.

Seleiman MF, Semida WM, Rady MM, Mohammad GF, Hemida KA, Alhammad BA, Hassan MM, Shami A (2020). Sequential application of antioxidants rectifies ion imbalance and strengthens antioxidant systems in salt-stressed cucumber. Plants 9(12):1783.

Seleiman M (2019). Use of plant nutrients in improving abiotic stress tolerance in wheat. In: Hasanuzzaman M, Nahar K, Hossain MA (Eds). Wheat Production in Changing Environments. Springer, Singapore, pp 481-495.

Slama I, xAbdelly A, Bouchereau A, Flowers T, Savoure A (2015). Diversity, distribution and roles of osmoprotective compounds accumulated in halophytes under abiotic stress. Annals of Botany 115:433-447.

Steel RGD, Torrie JH, Dickey D (1997). Principles and Procedures of statistics: a biometric approach. 3rd edition, McGraw-Hill Book Co., New York, USA pp 663-666.

Taamalli WL Youssef NB, Miled DDB, Zarrouk M (2004). Lipid breakdown in sunflower (Helianthus annuus L.) seeds during post germinative growth under salt-stress. Rivista Italian Delle Gostanze Grasse 81:90-97.

Taha RS, Seleiman MF, Alotaibi M, Alhammad BA, Rady MM, Mahdi AHA (2020). Exogenous potassium treatments elevate salt tolerance and performances of Glycine max L. by boosting antioxidant defense system under actual saline field conditions. Agronomy 10(11):1741.

Taha RS, Seleiman MF, Alhammad BA, Alkahtani J, Alwahibi MS, Mahdi AH (2021). Activated Yeast extract enhances growth, anatomical structure, and productivity of Lupinus termis L. plants under actual salinity conditions. Agronomy 11(1):74.

Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants: protective role of exogenous polyamines. Plant Science 151:59-66.

Yildirim E, Ekinci M, Turan M, Dursun A, Kul R, Parlakova F (2015). Roles of glycine betaine in mitigating deleterious effect of salt stress on lettuce (Lactuca sativa L.). Archives of Agronomy and Soil Science 61(12):1673-1689.

Zhu JK (2002). Salt and drought stress signal transduction in plants. Annual Review of Plant Biology 53(1):247-273.

Zhang XZ (1992) The measurement and mechanism of lipid peroxidation and SOD, POD and CAT activities in biological system. Research Methods of Crop Physiology 208-222.

How to Cite
DUSTGEER, Z., SELEIMAN, M. F., KHAN, I., CHATTHA, M. U., ALI, E. F., ALHAMMAD, B. A., JALAL, R. S., REFAY, Y., & HASSAN, M. U. (2021). Glycine-betaine induced salinity tolerance in maize by regulating the physiological attributes, antioxidant defense system and ionic homeostasis. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 49(1), 12248.
Research Articles
DOI: 10.15835/nbha49112248