Foliar applied proline and acetic acid improves growth and yield of wheat under salinity stress by improving photosynthetic pigments, physiological traits, antioxidant activities and nutrient uptake


  • Imran KHAN Department of Agronomy, University of Agriculture, Faisalabad, 38040 (PK)
  • Momina IQBAL Department of Agronomy, University of Agriculture, Faisalabad, 38040 (PK)
  • Athar MAHMOOD Department of Agronomy, University of Agriculture, Faisalabad, 38040 (PK)
  • Rizwan MAQBOOL Department of Agronomy, University of Agriculture, Faisalabad, 38040 (PK)
  • Muqarrab ALI Department of Agronomy, Muhammad Nawaz Shareef University of Agriculture, Multan (PK)
  • Muhammad T. ASLAM Department of Agronomy, University of Agriculture, Faisalabad, 38040 (PK)
  • Muhammad S. HANIF Fodder Research Institute Sargodha, Punjab (PK)
  • Shoaib A. KOHLI Fodder Research Institute Sargodha, Punjab (PK)
  • Sally NEGM Life Sciences Department, College of Science and Art, King Khalid University, Mohail, Aseer; Unit of Food Bacteriology, Central Laboratory of Food Hygiene, Ministry of Health, Sharkia (SA)
  • Mahmoud MOUSTAFA Department of Biology, College of Science, King Khalid University, Abha; Botany and Microbiology Department, Faculty of Science, South Valley University, Qena (SA)
  • Muhammad U. CHATTHA Department of Agronomy, University of Agriculture, Faisalabad, 38040 (PK)
  • Muhammad U. HASSAN Research Center on Ecological Sciences, Jiangxi Agricultural University, Nanchang, 330045 (CN)



antioxidants, growth, osmolytes, wheat, yield


Salinity stress (SS) is serious abiotic stress and a major limiting factor for crop productivity and global food security. In this context, the application of osmolytes is considered as an environmental friend approach to improve plant growth under SS. Thus, the present study was conducted to determine the impact of foliar applied proline (Pro) and acetic acid (AA) on growth, yield, physiological traits, photosynthetic pigments, ionic homeostasis and antioxidant activities of wheat under SS. The study contained SS levels 0, 6 and 12 dS m-1 and foliar spray of Pro and AA; water spray, Pro (75 mM), AA (15 mM) and AA (30 mM). The study was conducted in a completely randomized design with the factorial arrangement. Salinity stress significantly reduced wheat growth and yield, by decreasing relative water contents (-49.07%), photosynthetic pigments, free amino acids (FAA: -44.79%), total soluble proteins (TSP: -15.94%) and increasing the electrolyte leakage (EL: +27.28%), hydrogen peroxide (H2O2: +51.86%), and malondialdehyde (MDA: +36.91%) accumulation. The foliar spray of Pro and AA markedly improved the wheat growth and productivity through enhanced photosynthetic pigments, RWC, FAA, TSP, antioxidant activities (catalase: CAT, ascorbate peroxide: APX: peroxidase: POD), K+ and Ca2+ uptake and decreasing EL, MDA and H2O2 accumulation and restricted entry of toxic ions (Na+ and Cl-1).  Therefore, foliar application of AA and Pro effectively improves the growth and yield of wheat under SS by strengthening the antioxidant defense system, and maintaining ionic homeostasis and physiological performance.


Abogadallah GM (2010). Insights into the significance of antioxidative defense under salt stress. Plant Signaling and Behavior 5:369-374.

Al-Zahrani HS, Nahar K, Alharby HF, Alsamadany H, Hakeem KR, Hasanuzzaman M (2022). Zinc Supplementation Enhances Glutathione-Mediated Antioxidant Defense and Glyoxalase Systems to Conferring Salt Tolerance in Soybean (Glycine max L.). Agronomy 12:1032.

Alam R, Das D, Islam M, Murata Y, Hoque M (2016). Exogenous proline enhances nutrient uptake and confers tolerance to salt stress in maize (Zea mays L.). Progressive Agriculture 27:409-417.

Arnon D (1949). Copper enzyme in isolated chloroplast and chlorophyll expressed in terms of mg per gram. Plant Physiology 24:1-15.

Asada K (1987). Production and scavenging of active oxygen in photosynthesis. Photoinhibition 227-287.

Banakar MH, Amiri H, Ardakani MRS, Ranjbar GH (2022). Susceptibility and tolerance of fenugreek (Trigonella foenum-graceum L.) to salt stress: Physiological and biochemical inspections. Environmental and Experimental Botany 194:104748.

Bose J, Rodrigo-Moreno A, Shabala S (2014). ROS homeostasis in halophytes in the context of salinity stress tolerance. Journal of Experimental Botany 65:1241-1257.

Bradford M (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Annals of Biochemistry 72:248-54.

Chance B (1955). Assay of catalase and peroxidase. Methods in Enzymology 2:765-775.

Corwin DL (2021). Climate change impacts on soil salinity in agricultural areas. European Journal of Soil Science 72:842-862.

de Freitas PAF, de Souza Miranda R, Marques EC, Prisco JT, Gomes-Filho E (2018). Salt tolerance induced by exogenous proline in maize is related to low oxidative damage and favorable ionic homeostasis. Journal of Plant Growth Regulation 37:911-924.

Demiralay M, Altuntaş C, Sezgin A, Terzi R, Kadioğlu A (2017). Application of proline to root medium is more effective for amelioration of photosynthetic damages as compared to foliar spraying or seed soaking in maize seedlings under short-term drought. Turkish Journal of Biology 41:649-660.

Farooq M, Hussain M, Wakeel A, Siddique KH (2015). Salt stress in maize: effects, resistance mechanisms, and management. A review. Agronomy for Sustainable Development 35:461-481.

Hamilton P, Van Slyke D (1943). Amino acid determination with ninhydrin. Journal of Biology and Chemistry 150:231-250.

Hasanuzzaman M, Raihan MRH, Masud AAC, Rahman K, Nowroz F, Rahman M, Nahar K, Fujita M (2021). Regulation of reactive oxygen species and antioxidant defense in plants under salinity. International Journal of Molecular Sciences 22:9326.

Hashem A, Abd_Allah EF, Alqarawi AA, Al-Huqail AA, Wirth S, Egamberdieva D (2016). The interaction between arbuscular mycorrhizal fungi and endophytic bacteria enhances plant growth of Acacia gerrardii under salt stress. Frontiers in Microbiology 7:1089.

Hassan MU, Chattha MU, Ullah A, Khan I, Qadeer A, Aamer M, Khan AU, Nadeem F, Khan TA (2019). Agronomic biofortification to improve productivity and grain Zn concentration of bread wheat. International Journal of Agriculture and Biology 21:615-620.

Hu Y, Burucs Z, von Tucher S, Schmidhalter U (2007). Short-term effects of drought and salinity on mineral nutrient distribution along growing leaves of maize seedlings. Environmental and Experimental Botany 60:268-275.

Ikuyinminu E, Goñi O, O’Connell S (2022). Enhancing irrigation salinity stress tolerance and increasing yield in tomato using a precision engineered protein hydrolysate and Ascophyllum nodosum-derived biostimulant. Agronomy 12:809.

Iqbal MN, Rasheed R, Ashraf MY, Ashraf MA, Hussain I (2018). Exogenously applied zinc and copper mitigate salinity effect in maize (Zea mays L.) by improving key physiological and biochemical attributes. Environmental Science and Pollution Research 25:23883-23896.

Ivushkin K, Bartholomeus H, Bregt AK, Pulatov A, Kempen B, De Sousa L (2019). Global mapping of soil salinity change. Remote Sensing of Environment 231:111260.

Kim BM, Lee HJ, Song YH, Kim HJ (2021). Effect of salt stress on the growth, mineral contents, and metabolite profiles of spinach. Journal of the Science of Food and Agriculture 101:3787-3794.

Kim J-M, To TK, Matsui A, Tanoi K, Kobayashi NI, Matsuda F, Habu Y, Ogawa D, Sakamoto T, Matsunaga S (2017). Acetate-mediated novel survival strategy against drought in plants. Nature Plants 3:1-7.

Kosová K, Vítámvás P, Prášil IT, Renaut J (2011). Plant proteome changes under abiotic stress—contribution of proteomics studies to understanding plant stress response. Journal of Proteomics 74:1301-1322.

Ma X, Ou Y-B, Gao Y-F, Lutts S, Li T-T, Wang Y, Chen Y-F, Sun Y-F, Yao Y-A (2016). Moderate salt treatment alleviates ultraviolet-B radiation caused impairment in poplar plants. Scientific reports 6:1-15.

Machado RMA, Serralheiro RP (2017). Soil salinity: effect on vegetable crop growth. Management practices to prevent and mitigate soil salinization. Horticulturae 3:30.

Mahmood U, Hussain S, Hussain S, Ali B, Ashraf U, Zamir S, Al-Robai SA, Alzahrani FO, Hano C, El-Esawi MA (2021). Morpho-Physio-Biochemical and Molecular Responses of Maize Hybrids to Salinity and Waterlogging during Stress and Recovery Phase. Plants 10:1345.

Maimaiti A, Yunus Q, Iwanaga F, Mori N, Tanaka K, Yamanaka N (2014). Effects of salinity on growth, photosynthesis, inorganic and organic osmolyte accumulation in Elaeagnus oxycarpa seedlings. Acta Physiologiae Plantarum 36:881-892.

Mi J, Vallarino JG, Petřík I, Novák O, Correa SM, Chodasiewicz M, Havaux M, Rodriguez-Concepcion M, Al-Babili S, Fernie AR (2022). A manipulation of carotenoid metabolism influence biomass partitioning and fitness in tomato. Metabolic Engineering 70:166-180.

Miller G, Suzuki N, Ciftci‐Yilmaz S, Mittler R (2010). Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant, Cell and Environment 33:453-467.

Munns R, Gilliham M (2015). Salinity tolerance of crops–what is the cost? New phytologist 208:668-673.

Munns R, Tester M (2008). Mechanisms of salinity tolerance. Annual review of plant biology 59:651.

Nahar K, Hasanuzzaman M, Rahman A, Alam MM, Mahmud J-A, Suzuki T, Fujita M (2016). Polyamines confer salt tolerance in mung bean (Vigna radiata L.) by reducing sodium uptake, improving nutrient homeostasis, antioxidant defense, and methylglyoxal detoxification systems. Frontiers in Plant Science 7:1104.

Nakhaie A, Habibi G, Vaziri A (2020). Exogenous proline enhances salt tolerance in acclimated Aloe vera by modulating photosystem II efficiency and antioxidant defense. South African Journal of Botany.

Negrão S, Schmöckel S, Tester M (2017). Evaluating physiological responses of plants to salinity stress. Annals of Botany 119:1-11.

Pan J, Xue X, Huang C, Peng F, Liao J, Ma S, You Q, Wang T (2022). Salt tolerance strategies of Nitraria tangutorum Bobr. and Elaeagnus angustifolia Linn. determine the inoculation effects of microorganisms in saline soil conditions. Agronomy 12:913.

Parveen A, Liu W, Hussain S, Asghar J, Perveen S, Xiong Y (2019). Silicon priming regulates morpho-physiological growth and oxidative metabolism in maize under drought stress. Plants 8:431.

Parvin K, Nahar K, Hasanuzzaman M, Bhuyan MB, Mohsin SM, Fujita M (2020). Exogenous vanillic acid enhances salt tolerance of tomato: Insight into plant antioxidant defense and glyoxalase systems. Plant Physiology and Biochemistry 150:109-120.

Patade VY, Lokhande VH, Suprasanna P (2014). Exogenous application of proline alleviates salt induced oxidative stress more efficiently than glycine betaine in sugarcane cultured cells. Sugar Tech 16:22-29.

Prathap V, Ali K, Singh A, Vishwakarma C, Krishnan V, Chinnusamy V, Tyagi A (2019). Starch accumulation in rice grains subjected to drought during grain filling stage. Plant Physiology and Biochemistry 142:440-451.

Rady MM, Kuşvuran A, Alharby HF, Alzahrani Y, Kuşvuran S (2019). Pretreatment with proline or an organic bio-stimulant induces salt tolerance in wheat plants by improving antioxidant redox state and enzymatic activities and reducing the oxidative stress. Journal of Plant Growth Regulation 38:449-462.

Rady MM, Taha RS, Mahdi AH (2016). Proline enhances growth, productivity and anatomy of two varieties of Lupinus termis L. grown under salt stress. South African Journal of Botany 102:221-227.

Rahman M, Mostofa MG, Islam M, Keya SS, Das AK, Miah M, Kawser A, Ahsan S, Hashem A, Tabassum B (2019). Acetic acid: A cost-effective agent for mitigation of seawater-induced salt toxicity in mung bean. Scientific Reports 9:1-15.

Rahman M, Mostofa MG, Keya SS, Rahman A, Das AK, Islam R, Abdelrahman M, Bhuiyan SU, Naznin T, Ansary MU (2021). Acetic acid improves drought acclimation in soybean: an integrative response of photosynthesis, osmoregulation, mineral uptake and antioxidant defense. Physiologia plantarum 172:334-350.

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

Sangwongchai W, Krusong K, Thitisaksakul M (2022). Salt tolerance at vegetative stage is partially associated with changes in grain quality and starch physicochemical properties of rice exposed to salinity stress at reproductive stage. J. Sci. Food Agric. 102: 370-382.

Saidi S, Cherif-Silini H, Chenari Bouket A, Silini A, Eshelli M, Luptakova L, Alenezi FN, Belbahri L (2021). Improvement of Medicago sativa crops productivity by the co-inoculation of Sinorhizobium meliloti–Actinobacteria under salt stress. Current Microbiology 78:1344-1357.

Seleiman MF, Aslam MT, Alhammad BA, Hassan MU, Maqbool R, Chattha MU, Khan I, Gitari HI, Uslu OS, Rana R (2022). Salinity stress in wheat: Effects, mechanisms and management strategies. Phyton 91:667.

Shahid SA, Zaman M, Heng L (2018). Soil salinity: Historical perspectives and a world overview of the problem. In: Guideline for salinity assessment, mitigation and adaptation using nuclear and related techniques. pp 43-53. Springer.

Steel R, Torrie J, Dickey M (1980). A biometrical approach. Principles and Procedures of Statistics 8-566.

Taïbi K, Taïbi F, Abderrahim LA, Ennajah A, Belkhodja M, Mulet JM (2016). Effect of salt stress on growth, chlorophyll content, lipid peroxidation and antioxidant defence systems in Phaseolus vulgaris L. South African Journal of Botany 105:306-312.

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.

Yang S-L, Lan S-S, Gong M (2009). Hydrogen peroxide-induced proline and metabolic pathway of its accumulation in maize seedlings. Journal of Plant Physiology 166:1694-1699.

Yildiz M, Terzi H (2013). Effect of NaCl stress on chlorophyll biosynthesis, proline, lipid peroxidation and antioxidative enzymes in leaves of salt-tolerant and salt-sensitive barley cultivars. Journal of Agricultural Sciences 19:79-88.

Zali AG, Ehsanzadeh P (2018). Exogenous proline improves osmoregulation, physiological functions, essential oil, and seed yield of fennel. Industrial Crops and Products 111:133-140.

Zhang J, Zhang Q, Xing J, Li H, Miao J, Xu B (2021). Acetic acid mitigated salt stress by alleviating ionic and oxidative damages and regulating hormone metabolism in perennial ryegrass (Lolium perenne L.). Grass Research 1:1-10. G

Zhao C, Zhang H, Song C, Zhu J-K, Shabala S (2020). Mechanisms of plant responses and adaptation to soil salinity. The innovation 1:100017.

Zörb C, Geilfus CM, Dietz KJ (2019). Salinity and crop yield. Plant Biology 21:31-38.



How to Cite

KHAN, I., IQBAL, M., MAHMOOD, A., MAQBOOL, R., ALI, M., ASLAM, M. T., HANIF, M. S., KOHLI, S. A., NEGM, S., MOUSTAFA, M., CHATTHA, M. U., & HASSAN, M. U. (2022). Foliar applied proline and acetic acid improves growth and yield of wheat under salinity stress by improving photosynthetic pigments, physiological traits, antioxidant activities and nutrient uptake. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 50(3), 12820.



Research Articles
DOI: 10.15835/nbha50312820

Most read articles by the same author(s)

1 2 > >>