Differential Modulation of Antioxidant Defense and Salinity Tolerance in Maize by Selenium, α-Tocopherol, and Ascorbic Acid

Differential Modulation of Antioxidant Defense and Salinity Tolerance in Maize by Selenium, α-Tocopherol, and Ascorbic Acid | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Differential Modulation of Antioxidant Defense and Salinity Tolerance in Maize by Selenium, α-Tocopherol, and Ascorbic Acid Radwan Khalil, Amina Gamal, Mohammad Yusuf, Samia Haroun, Metwally Rashad, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8692316/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 12 You are reading this latest preprint version Abstract Selenium, α-tocopherol, and ascorbic acid are important antioxidants that mitigate abiotic stresses in plants, yet their comparative effects under salinity remain insufficiently studied. This study aimed to evaluate and compare the effects of selenium, α-tocopherol, and ascorbic acid on growth, carbon metabolism, and ion uptake in salt-stressed maize plant. Deionised water (control), selenium (0.5 mM), α-tocopherol (200 ppm), or ascorbic acid (500 ppm) were applied to maize seeds, which were then cultivated at NaCl concentrations of 0, 100, 150, and 200 mM. Plant samples were examined for ionic, physiological and biochemical characteristics after 40 days. Salt stress caused concentration-dependent reductions in growth, chlorophyll, insoluble sugars, carbohydrates, phenolics, flavonoids, and ion uptake, while enhancing soluble sugars, α-amylase activity, and sodium accumulation. On the other hand, growth performance, pigment content, carbohydrate and secondary metabolites, and the K⁺/Na⁺ and Ca²⁺/Na⁺ ratios were all improved by antioxidant treatments administered in non-stressful environments. Among the tested compounds, ascorbic acid was most effective in alleviating salt-induced damage. It enhanced antioxidant activity, improved carbon metabolism and ion homeostasis, and induced proteins linked to salt tolerance, leading to better growth under salinity. In conclusion, ascorbic acid shows promise as a way to reduce salt stress and increase maize yields in salty soils. Biological sciences/Biochemistry Biological sciences/Physiology Biological sciences/Plant sciences Selenium α-tocopherol Ascorbic acid Salt stress Protein Antioxidants Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Crop production has not kept up with the growing need for food, and global agriculture is today confronted with the daunting task of producing almost 70% more food to fulfil the demands of a fast expanding population. Over 6% of the world’s arable land is affected by salinity (Parvaiz et al., 2008), with saline soils predominantly distributed across arid and semi-arid regions of Asia (e.g., South and Central Asia, the Middle East), Africa (notably North Africa), Australia, and parts of the Mediterranean basin, as well as irrigated agricultural lands in North and South America. Although several of these regions are not traditionally major maize growing areas, maize was selected for this study because of its global economic importance, its expanding cultivation into marginal and saline-prone environments due to climate change and irrigation-induced salinization, and its well-documented sensitivity to salt stress. Consequently, maize serves as an appropriate model crop for evaluating stress-mitigation strategies aimed at improving crop resilience under saline conditions. This issue is getting worse all across the world, mostly because of the usage of subpar irrigation water and insufficient drainage infrastructure. Salt stress negatively impacts subsequent plant growth and developmental processes in addition to seed germination (Nakashima et al., 2000; Dash and Panda, 2001). Excessive salt accumulation leads to ionic imbalance and osmotic stress, which together disrupt cellular homeostasis and can ultimately result in plant death (Yu and Gu 2013; Gong et al. 2018 ). Furthermore, excess of Cl + and Na + ions buildup in tissues of plants caused by stress caused by salt interferes with vital physiological and metabolic functions and upsets normal ionic homeostasis (Arif et al. 2020 ) The cellular Na + /K + ratio, which is essential for preserving membrane potential and enzymatic activity, is also disrupted by this imbalance. Plants have, of course, created a variety of adaptive techniques to lessen the adverse consequences of various abiotic stressors. Although it can be hazardous to the environment, selenium serves as a necessary trace element for both people and animals. Its beneficial and harmful effects are closely separated and rely on a number of variables, including its chemical form, concentration, and environmental circumstances (Yu and Gu, 2013). Humans, plants, animals, and microbes all contain trace levels of selenium. Despite being needed in very small amounts, it is an essential element that has been demonstrated to increase a plant's resilience to oxidative damage brought on by UV light, postpone senescence, and encourage the growth and vitality of ageing seedlings (Rostami and Abbaspour 2019). The low-molecular-weight lipophilic antioxidant α-tocopherol, also referred to as vitamin E, is produced spontaneously by organisms that use green photosynthesis (Munné-Bosch and Alegre 2000; Munné-Bosch 2005 ; Munné-Bosch 2007 ; Falk and Munné-Bosch 2010). Plant membranes are the primary location for both α- and γ-tocopherol. In particular, α-tocopherol is the primary type found in plant leaves' membranes involved in photosynthetic processes, while γ-tocopherol is mostly found in tissues that are not photosynthetic, such as nuts, seeds, and fruits (Grilo et al. 2014 ). Although it may also occur, to a lesser extent, in other cellular compartments including the cytoplasm and vacuoles, tocopherol production mostly takes place within chloroplasts—more precisely, in the plastoglobuli, thylakoid membranes, and envelope (Rautenkranz et al. 1994 ; Li et al. 2008) α-Tocopherol is essential for shielding plants from cellular oxidation brought on by stress. Exogenous α-tocopherol injection has been demonstrated to successfully enhance plant growth and developmental processes across a range of adverse environmental conditions (Sadiq, Akram, and Ashraf 2017; Khalil et al. 2021 ). Because it is prevalent in cellular membranes, α-tocopherol prevents lipid peroxidation and scavenges reactive free radicals, preserving membrane integrity under stress (Munné-Bosch 2007 ). Tocopherols not only shield cell membranes but also control the production of particular genes that respond to stress, which helps to modify plant defence and adaption systems (Munné-Bosch 2007 ; Li et al. 2008; Suo et al. 2017 ). Ascorbic acid, commonly referred to as vitamin C, is a water-soluble antioxidant, is an essential substrate for the enzymatic detoxification of hydrogen peroxide. By controlling cell development and division, it encourages plant growth and improves resistance to abiotic challenges including salinity and temperature extremes (Smirnoff 1995 ). By increasing plant resistance to salinity, drought, and pests, seed priming improves crop performance in stressful environmental conditions (Harris et al. 2007 ). This method eventually increases output by assisting crop plants in overcoming stressors like drought and pest damage ( Harris et al. 2007 ). According to (David Harris et al. 2008 ) The nutrient content of seeds is improved by nutrient seed priming, germination as well as seedling establishment stressful by soaking seeds in nutrient solutions rather than water. This study aims to elucidate the potential role of seed priming with ascorbic acid, α-tocopherol, and selenium in enhancing salinity tolerance of maize. The research explores how these treatments modulate plant growth, physiological attributes, antioxidant defense mechanisms, and nutrient homeostasis under varying salinity levels, ultimately contributing to improved crop performance and yield stability under salt stress conditions. Materials and techniques Plant material We purchased pure Seeds of maize ( Zea mays L.) (TWC 321) from the Ministry of Agriculture's Agricultural Research Centre in Giza, Egypt. Chemical source and its preparation Sigma–Aldrich (USA) provided the sodium chloride (NaCl), ascorbic acid, sodium selenium, and α-tocopherol. Based on initial tests, working solutions of ascorbic acid (250 ppm), α-tocopherol (200 ppm), and sodium selenate (0.5 µM) were made. In order to create salinity stress, sodium chloride was employed as the salt source at concentrations of 100, 150, and 200 mM. Design of experiments and treatment regimens The study was conducted in a greenhouse at Benha University's Faculty of Science's Botany and Microbiology Department in Egypt, using 80 plastic pots (25 × 40 cm) filled with a clay–sand combination (2:1, v/v). Five replicates of a completely randomised block design were used for the treatments. Five seeds per pot were planted after surface-sterilized Zea mays L. seeds were soaked for 12 hours in either sodium selenate (0.5 µM), α-tocopherol (200 ppm), ascorbic acid (250 ppm), or deionised water (control). With soil moisture kept at 80% field capacity, Plants were exposed to 0, 100, 150, and 200 mM NaCl at 21 days. Samples for growth, physiological, biochemical, and SDS-PAGE protein analysis were gathered at 40 days. Growth characteristics Samples of plants were split into roots, stems, and leaves after being carefully cleaned with distilled water to get rid of any remaining soil particles. Recorded were the main root and stem lengths, each plant's leaf count and leaf area (cm²). Following the measurement of the fresh weights of the roots and stems, every sample was dried in an oven at 70°C in order to calculate their dry weight (g/plant). For ensuing biochemical investigations, tissues that had been dried were pulverised into a fine powder and stored dry. Identifying and extracting photosynthetic pigments The process described by was used to extract and find out the total amount of photosynthetic pigments Fadeel ( 1962 ). Anhydrous 100% acetone was used to remove a known fresh weight of the topmost completely inflated leaves, which were then stored at -5°C for at least 12 hours. Using cold acetone and acid-washed sand in a mortar and pestle, the tissues were ground into a homogeneous mixture on ice. Anhydrous sodium sulphate was added to help with dehydration during grinding, and a tiny bit of magnesium carbonate was added to balance the acidity of the tissue. After 15 minutes of centrifuging the homogenate at 4000 × g, the residue was extracted again using cold acetone to remove all of the pigment. 100% acetone was used to make up the combined extracts to a given volume. A Perkin-Elmer double-beam spectrophotometer (Model 200–20) was used to detect absorbance at 440.5, 644, and 662 nm. The amounts of carotenoid, chlorophyll a, and chlorophyll b were determined using conventional formulas for extracts made entirely of acetone (Sestak et al ., 1971): Chlorophyll a = 9.78 E662- 0.99 E644 (µg /ml) Chlorophyll b = 21.4 E644–4.65 E662 (µg /ml) Chlorophyll (a + b) = 5.13 E662 + 20.41 E644 (µg /ml) Carotenoids = 4.69 E440.5 − 0.268 chlorophyll (a + b) (µg /ml) The outcomes of the aforementioned equations were converted to mg pigment g⁻¹ fresh weight after being expressed as µg pigment ml⁻¹ extracts (Fadeel, 1962 ). Stress biomarkers (electrolyte leakage (EL), membrane stability index (MSI) lipid-peroxidation (Malondialdehyde; MDA) The technique outlined by was used to measure the total amount of inorganic ions that seeped out of the leaves Sullivan ( 1979 ). Leaf discs were boiled at 100°C (EC𝚌) after being incubated in 10 ml of deionised water at 45°C (ECₐ) and 55°C (EC β ) for 30 minutes in order to quantify electrolyte leakage. The calculation for leakage (%) was: Electrolyte leakage (%) = (ECb-ECa)/ECcX100 where, EC = Electrical Conductivity. The MSI was evaluated by incubating two sets of 200 mg of leaf tissue in 10 ml of deionised water. For 30 minutes (C₁), the initial set was heated to 40°C, and for 10 minutes (C₂), boiling the second set was done at 100°C. A conductivity meter was used to record electrical conductivities. MSI was computed using the formula outlined by (Sairam, 1994 ). MSI (%) = [1 – (C1/C2) × 100] The degree of lipid peroxidation was measured using the method of malondialdehyde (MDA) content Hodges et al. ( 1999 ). After homogenising two grammes of leaf tissue in ten millilitres of 5% thiobarbituric acid (TBA), centrifuging the mixture for ten minutes at 15,000 × g. 4ml of 0.5% TBA in 20% trichloroacetic acid (TCA) were added to two millilitres of the supernatant. The mixture was rapidly chilled in an ice bath after being heated to 95°C for 30 minutes and centrifuged once more for 10 minutes at 10,000 × g. Non-specific absorbance at 600 nm was deducted from the supernatant's absorbance, which was measured at 532 nm. To calculate the MDA concentration, an extinction coefficient of 155 mM⁻¹ cm⁻¹ was employed, which was then reported as µg MDA g⁻¹ fresh weight. Measurement and extraction of proline The calculation of free proline was done using (Bates et al., 1973 ). It was fresh tissue homogenised in Sulfosalicylic acid (3%), the filtrate was reacted with Ninhydrin acid, and toluene was used to extract the amount of proline. Proline concentration was established using a standard curve and represented as mg proline 100 g⁻¹ dry weights after absorbance was measured at 520 nm. Phenolic compounds' extraction and identification The process of extracting phenolic compounds followed the protocol described by Sauvesty et al. ( 1992 ). 70% ethanol was used to extract phenolic chemicals from dry powdered tissue over the course of an overnight period at 40°C. After centrifuging and low-pressure evaporation, the extract was diluted with distilled water. Applying the Folin-Ciocalteu technique, one millilitre of The sample was hydrolysed with 2 N HCl to liberate bound phenolics, neutralised, and utilised to determine the total amount of phenol (Lowe, 1993 ) was employed to determine the phenolic aglycone. Flavonoid extraction and identification Aluminium chloride colorimetric analysis was employed to calculate the total flavonoid concentration (Sakanaka et al. , 2005 ). Methanol was utilised to extract the dried materials, and 0.25 millilitres of the clear extract were added to 1.25 millilitres of distilled water. Following the addition of 75 µl of a solution containing 5% sodium nitrite, the mixture was incubated for six minutes. After adding 150 µl of a 10% aluminium chloride solution and letting it stand for five minutes, 0.5 ml of 1 M sodium hydroxide was added. The absorbance was measured at 510 nm after the total volume was adjusted to 2.5 ml using distilled water. To calculate the total flavonoid content, a myricetin standard curve was utilised, which was then reported as mg g⁻¹ dry weight. Protein extraction and identification The method described by was used to extract proteins Eltayeb et al. ( 2007 ). To extract the soluble protein, 100 mg of dry powdered tissue was incubated for two hours at 90°C in 10 ml of H 2 O. The mixture was centrifuged for 10 minutes at 5000 g after cooling. After that, the leftover residue was homogenised with 10 millilitres of 1 N NaOH for two hours at 90°C in order to remove the water-insoluble protein. Following a 10-minute centrifugation at 5000 g, HCl was used to neutralise the clear supernatant, and distilled water was added to each extract to reach a given volume. Protein analysis was done using the modified Folin-Lowry technique that was used by Hartree ( 1972). After combining one millilitre of the transparent extract of protein with 0.9 millilitres of alkaline solution of sodium carbonate, the mixture was heated to 50°C for ten minutes. Following cooling, a solution of potassium sodium tartrate and copper sulphate was added in 0.1 millilitres, and the mixture was allowed to stand at room temperature for ten minutes. The 10% Folin-Ciocalteu reagent (3 ml) was then added and mixed right away. Protein concentration was determined using a bovine serum albumin standard curve and represented as mg g⁻¹ dry weight after 30 minutes, when absorbance was measured at 750 nm against a reagent blank. Protease and Amylase assays The fresh tissue was homogenised in a pH 7.6 phosphate buffer containing 20 mM to extract protease, and the buffer used was 100 mM acetate (pH 6.0) to extract amylase. It made use of bovine serum albumin (BSA) as the substrate to measure protease activity. After being incubated at 40°C for 60 minutes, the reaction mixture which contained crude extract and BSA solution—was terminated with 10% trichloroacetic acid. Following centrifugation, the modified Folin–Lowry method was used to quantify the amount of soluble peptides Hartree ( 1972 ). In order to measure amylase activity, half a millilitre of crude extract and half a millilitre of 0.5% soluble starch were incubated for 30 minutes at 40°C in 0.1 M acetate buffer (pH 6.0) with 5 mM CaCl₂. HgCl₂ was used to stop the reaction, and Nelson's technique was used to measure the reducing sugars (Clark and Switzer, 1977 ). Antioxidant enzyme extraction and analysis A 100 mM phosphate buffer (pH 7) containing 0.1 g PVP and 0.1 mM Na₂EDTA was used to homogenise 0.25 g of fresh leaf material in liquid nitrogen. After centrifuging the homogenate for 10 minutes at 15,000 × g, the supernatant was centrifuged again for 10 minutes at 18,000 × g. The final extract was gathered and maintained at 4°C to gauge catalase (CAT) and peroxidase (POD) activity. Peroxidase (POX) activity Guaiacol was used as the substrate to assess the activity of peroxidase (POX, EC 1.11.1.7). 10 mM phosphate buffer (pH 7.0), 10 mM H 2 O₂, 20 mM guaiacol, and 0.5 ml of the crude enzyme extract were all included in the reaction mixture (Malick and Singh, 1980 ). The increase in absorbance at 470 nm was used to track the production of oxidised guaiacol (Klapheck et al., 1990) use an ultraviolet spectrophotometer (Spectronic 601). The change in optical density g-1 fresh weight min-1 is used to express enzyme activity. Catalase (CAT) activity 0.5 millilitres of enzyme extract in the reaction mixture, 30% (w/v) H 2 O 2 , and phosphate buffer (50 mM, pH 7.0) was used to measure CAT activity (EC 1.11.1.6.) (Aebi, 1984 ). Catalase activity was measured using a Spectronic 601 UV spectrophotometer as the decrease in absorbance at 240 nm caused by H 2 O₂ breakdown and expressed as µM H 2 O 2 oxidised g − 1 fresh weight min- 1 (Havir and McHale 1987). Phenylalanine ammonia-lyase (PAL) determination PAL (EC 4.3.1.5) extraction and assay were performed as instructed Solecka and Kacperska (2003). Following a 20-minute centrifugation at 12,000 × g at 4°C, fresh leaf tissue was homogenised in liquid nitrogen using extraction buffer (5 mM EDTA; 5 mM ascorbic acid; 50 mM Tris–HCl, pH 8.9). The PAL assay used the supernatant as a crude enzyme. 100 µl of enzyme extract and 0.9 ml of substrate solution (16 mM L-phenylalanine in 50 mM Tris–HCl, pH 8.9, with 3.6 mM NaCl) were included in the reaction mixture. It was incubated for one hour at 37°C and stopped with 500 µl of 6 N HCl. At 290 nm, absorbance was measured. nmol trans-cinnamic acid g⁻¹ FW h⁻¹ was used to express PAL activity, which is the quantity that produces 1 µM trans-cinnamic acid every hour. Determination of polyphenol oxidase (PPO) Polyphenol oxidase (EC 1.14.18.1) was extracted as described by Kar and Mishra (1976) with slight modification. After being homogenised in ice-cold 0.1 M phosphate buffer (pH 7.0) at 4°C, Samples of frozen tissue were centrifuged at 10,000 × g for 10 minutes. Polyphenol oxidase (PPO) activity was measured using the resultant supernatant using the procedure of Nguyen et al. ( 2003 ). 2.5 ml of substrate solution (0.05 M phosphate buffer) and 0.5 ml of crude enzyme extract, pH 6.0, with Catechol at 0.05 M) made up the assay combination, which was then incubated for 30 minutes at 30°C. PPO activity was reported as ABS unit fresh weight h⁻¹ g⁻¹, and absorbance was measured at 420 nm. Protein pattern analysis by gel electrophoresis SDS-PAGE was employed to separate protein samples on a 12% polyacrylamide gel. According to their size, negatively charged proteins in this technology move through the gel matrix in the direction of the anode under an electric field by using the Laemmli ( 1970 ) as edited by Studier ( 1973 ) and equal amounts of isolated proteins were loaded. Gel running and staining After combining protein samples with an equivalent amount of sample buffer, they were denatured for three to five minutes at 80 to 90 degrees Celsius, cooled on ice, and then placed onto the gel. Using 1× Tris/glycine–SDS running buffer, electrophoresis was carried out at 80 V for the stacking gel and 100 V for the separating gel. Detection of protein bands SDS-PAGE 200 ml of the gels was stained overnight with Coomassie Brilliant Blue R-250, and the distaining solution (200 ml) was utilised to distain them for two hours while gently shaking them. This process was repeated until the background was clear. Using Gel Pro Analyser version 3 software, separated protein bands were scanned and examined using a Gel Documentation System (GDS) to compare molecular weights, band intensity, and polypeptide patterns to common protein markers. Extraction and estimation of certain minerals: In an oven set to 80˚C, plant materials were dried until their weight remained constant. The dry material was broken down using the technique of Chapman and Pratt (1962). A 250 ml flask for digestion that had been cleaned with acid was filled with a 0.2 g sample of ground plant material. On a heater, the sample was broken down till dense white fumes formed and the solution cleared to around 2.5 millilitres when six millilitres of a concentrated sulfuric–perchloric acid mixture (70%, 5:1 v/v) was added. A 50 ml volumetric flask was used to quantitatively transfer the digest once it had cooled. It was then diluted to volume using Whatman No. 42 paper was used to filter the distilled water. The filtrate was kept for K, Ca, Na, Mg, and Se analyses. Using a flame photometer, potassium was measured by Ranganna (1977) and atomic absorption of Ca, Na, mg, and Se using the method described in Julshamn et al . ( 2005 ) Statistical Analysis IBM SPSS Statistics for Windows, Version 19.0, was used to statistically analyse the experiment's data using analysis of variance (ANOVA). IBM Corp., Armonk, NY, and displayed as treatment mean ± SE. The least significant difference (LSD) at p ≤ 0.05 was used to compare the treatment means. The LSD test indicates that bars with the same letter are not substantially different at p < 0.05. Results Growth performance When maize plants were exposed to salt stress and those that were not, their responses to three distinct growth regulators (selenium, α-tocopherol, and ascorbic acid) varied in terms of their growth performance (length of shoot and root; fresh mass of shoot and root; dry mass of shoot and root; leaf area; Figs. 1A-F and Fig. 2A). When compared to the untreated (water-soaked) control plants, the pre-sowing seed soaking treatment of Selenium, α-tocopherol, and ascorbic acid significantly increased the following attributes: shoot length (4.4, 8.5, and 8.97%), root length 13.9, 14.3 and 16.19%), shoot fresh mass (1.7, 11.7 and 12.52%), root fresh mass (66.6, 67.1 and 69.33%), shoot dry mass (21.1, 42.1 and 89.47%), root dry mass (24, 52, and 56%), and leaf area (9.2, 25.5 and 27.7%). However, in a concentration-dependent manner, growth performance was dramatically reduced by increasing salt concentrations (0, 50, 100, and 200 mM). In contrast to plants treated with selenium and α-tocopherol, ascorbic acid significantly restored the loss, although seed soaking treatments of selenium and α-tocopherol, and ascorbic acid under salt stress were able to overcome the damage. Plants treated with ascorbic acid through pre-sowing seed soaking treatment demonstrated the highest growth performance of maize plants under salt stress. Figure 1. Effect of sodium selenate (Na 2 SeO 4 ), a-tocopherol, and ascorbic acid on the morphological traits (a) shoot length, (b) root length, (c) shoot fresh mass, (d) root fresh mass, (e) shoot dry mass, and (f) root dry mass of 40 days old maize ( Zea mays L.) plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test. Pigments and Photosynthetic performance When compared to control plants, maize plants examined 40 days after sowing showed a substantial drop in chlorophyll a, b, and carotenoid levels at increasing salt concentrations (0, 50, 100, and 200 mM) (Figs. 2B-D). When compared to non-salinized control plants, the largest inhibitory effect of salinity on carotenoid (67%), chlorophyll a (77.1%), and chlorophyll b (64%), was observed at 200 mM NaCl. However, as compared to untreated control plants, treatments with selenium, α-tocopherol, and ascorbic acid considerably enhanced the quantities of photosynthetic pigments. They also recovered the loss caused by the same concentrations of salt in maize plants. Furthermore, ascorbic acid was found to be more promising and to have the greatest protective impact against all concentrations of salt stress in maize plants when compared to selenium, α-tocopherol, and ascorbic acid. Figure 2. Effect of sodium selenate (Na 2 SeO 4 ), a-tocopherol, and ascorbic acid on (a) leaf area per plant, (b) chlorophyll a, (c) chlorophyll b, and (d) carotenoids of 40 days old maize ( Zea mays L.) plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test. Stress Biomarkers [Electrolyte leakage (EL), Membrane stability index (MSI), Lipid peroxidation and proline] Lipid peroxidation and EL significantly increased when concentrations of NaCl (0, 50, 100, and 200 mM) were applied compared to the control plants. When compared to control plants, the plants exposed to 200 mM NaCl showed the highest levels of electrolyte leakage and lipid peroxidation, which were 50.4% and 104.2%, respectively (Figs. 3 A- 3 C). However, when selenium, α-tocopherol, and ascorbic acid were present, maize plants under NaCl stress shown a considerable decrease in electrolyte leakage of 42%, 48.6%, and 53.3%, respectively. Furthermore, when compared to maize plants exposed to 200 mM salt stress, lipid peroxidation also demonstrated a significant decrease of 54.7%, 69.1%, and 79.4% in the presence of selenium, α-tocopherol, and ascorbic acid, respectively. As maize plants were subjected to 200 mM NaCl, their MSI dropped by 29.4% as compared to control plants (Figs, 3B). However, when tested on plants under NaCl stress, ascorbic acid recovered the loss of MSI by 40.2% compared to plants subjected to 200 mM of NaCl, out of selenium, α-tocopherol, and ascorbic acid. Salinity stress significantly increased proline accumulation in maize shoots, with a progressive rise at 100–200 mM NaCl and a maximum increase of 24.2% at 200 mM compared with non-salinized plants (Figs, 3D). Exogenous application of sodium selenate, α-tocopherol, or ascorbic acid further enhanced proline content under salinity. At 100 mM NaCl, proline increased by 23.1%, 18.18%, and 31.81%, respectively, relative to the salinity-stressed control, indicating an enhanced osmotic adjustment under salt stress Nutritional status (Carbohydrates, Phenolics, Flavonoids, and Protein) Figures 4A–D make it clear that, when compared to control plants, maize plants displayed a significant pattern of change for protein, phenolics, carbohydrates, and flavonoids regardless of treatment patterns. The nutritional status of maize plants was significantly reduced by all applied concentrations of NaCl (0, 50, 100, and 200 mM); however, plants exposed to 200 mM salt stress showed the greatest decreases in carbohydrates (20.03%), phenolics (27.6%), flavonoids (37.5%), and protein (35.3%) when compared to non-salinized control plants. Furthermore, as compared to their respective control or salinized plants, the application of selenium, α-tocopherol, and ascorbic acid through seed soaking enhanced the amounts of total carbohydrates, phenolics, flavonoids, and protein in the maize plant's shoot. Figure 4. Effect of sodium selenate (Na 2 SeO 4 ), a-tocopherol, and ascorbic acid on (a) total carbohydrate content, (b) total phenolic content, (c) total flavonoid content, and (d) total protein content of 40 days old maize ( Zea maysL.) plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test. Antioxidant enzymes Peroxidase and Catalase, (Amylases, Proteases, Phenylalanine ammonia- lyase, Polyphenol oxidase activities) When sodium selenium, α-tocopherol, and ascorbic acid were applied through seeds, the activity of amylases, proteases, and peroxidases was greatly reduced; however, the activities of phenylalanine ammonia-lyase, polyphenol oxidase, and catalase increased in comparison to the plants that were not treated and the plants that were treated with NaCl (Figs. 5 A-F). However, in maize plants, the activities of amylases, proteases, polyphenol oxidase, and peroxidase were all stimulated by salt stress. When compared to the untreated plants, the 200 mM salt treatment had the greatest stimulatory effect on amylases, protease, polyphenol oxidase, and peroxidase activities, increasing them by 60.4%, 64.3%, 58.97%, and 38.14%, respectively. Phenylalanine ammonia-lyase and catalase also decreased as the concentration of salt increased. Protein Profile SDS-PAGE analysis of maize leaves revealed protein bands ranging from 95.54 to 39.28 kDa across all treatments. In both untreated and salt-stressed plants, three protein bands (95.54, 72.02, and 44.63 kDa) were consistently expressed, regardless of sodium selenate or α-tocopherol application, suggesting these may represent constitutive or housekeeping proteins. Control plants exhibited six bands (95.54, 72.02, 67.93, 59.29, 44.63, and 39.28 kDa), whereas salt stress alone altered the pattern to four bands (95.54, 72.02, 59.29, and 44.63 kDa), indicating the possible downregulation or degradation of specific stress-sensitive proteins. Treatment with sodium selenate or α-tocopherol induced the appearance of a new 82.84 kDa band in control plants, suggesting activation of stress-protective proteins, whereas in salt-stressed plants, two bands (59.29 and 39.28 kDa) appeared transiently and then disappeared, which may reflect dynamic protein regulation under combined stress and treatment conditions. Seed priming with 250 ppm ascorbic acid increased the number of protein bands in plants exposed to 100–200 mM NaCl, indicating enhanced synthesis of stress-responsive proteins. Notably, ascorbic acid treatment led to the de novo synthesis of two polypeptides (80.47 and 67.43 kDa) compared to controls, which displayed three bands (80.47, 67.43, and 34.93 kDa) that disappeared under salt stress. These changes suggest that ascorbic acid may stimulate protective protein expression, potentially including antioxidant enzymes or osmoprotective proteins, enhancing the plant’s tolerance to salinity. Mineral contents In comparison to control plants, the accumulation of Na+ was 33.3% at the maximum salinity (200 mM NaCl). However, as NaCl concentrations increased, the amounts of K + , Ca +2 , K + /Na + , Ca +2 /Na + , and Mg +2 in maize shoots dropped dramatically, reaching their lowest values in plants treated with 200 mM. These contents were estimated to be 9.25%, 9.79%, 27.20%, 27.63%, and 10.77% lower than those of their respective control plants (Table 1). Ascorbic acid, sodium selenate, and α-tocopherol significantly altered the levels of Na + , K + , Ca +2 , K + /Na + , Ca +2 /Na + , and Mg+2 when compared to control plants and plants treated to varying concentrations of NaCl. Conversely, ascorbic-soaked seed exhibited the greatest impact on the levels of inorganic cations (Na + , K + , Ca +2 , K + /Na + , Ca +2 /Na + , and Mg +2 ). In comparison to untreated control plants and 200 mM salinized plants, Na + decreased by 38.9% and 19.81%, respectively, while K + , Ca +2 , K + /Na + , Ca +2 /Na + , and Mg +2 increased by 31.4%, 43.5%, 108.5%, 127.9%, and 96.2%, respectively, when compared to reference control, and increased by 15.3%, 26.8%, 43.8%, 58.3%, and 57.1% %, respectively, in comparison to plants treated with 200 mM NaCl. Table 1. Effect of sodium selenate (Na2SeO4), a-tocopherol, and ascorbic acid on uptake of (a) Na + , (b) K + , (c) Ca + , (d) K + /Na + , (e) Ca 2+ /Na 2+ , and (f) Mg 2+ of 40 days old maize (Zea mays L.) plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test Stage Treatments Nacl mM Na + % K + % Ca +2 % K + /Na + % Ca +2 /Na + % Mg +2 % 40-day- old maize plant Reference control 0 2.720±0.001 i 1.882±0.001 g 2.245±0.002 g 0.692±0.001 g 0.825±0.003 g 1.958±0.002 e 100 2.805±0.003 j 1.798±0.003 d 2.237±0.002 d 0.640±0.003 f 0.797±0.001 f 1.871±0.006 d 150 3.371±0.005 o 1.694±0.001 a 2.111±0.001 a 0.502±0.001 a 0.626±0.004 b 1.801±0.004 c 200 3.387±0.002 p 1.708±0.002 b 2.025±0.001 b 0.504±0.000 a 0.597±0.002 a 1.747±0.001 a sodium selenate 0.5 µM 0 2.293±0.002 e 1.891±0.003 h 2.746±0.004 h 0.824±0.002k 1.197±0.002 l 2.480±0.002 j 100 2.371±0.001 f 1.810±0.002 e 2.647±0.004 e 0.763±0.003 i 1.116±0.001 j 2.410±0.001 h 150 3.187±0.002 l 1.76±0.001 c 2.511±.0.002 c 0.551±0.001 c 0.787±0.003 e 2.088±0.003 f 200 3.347±0.002 n 1.802±0.003 d 2.187±0.001 d 0.538±0.004 b 0.653±0.006 c 1.781±0.001 b α-tocopherol 200ppm 0 2.137±0.04 c 2.265±0.005 m 3.163±0.005 m 1.059±0.001 m 1.480±0.008 n 3.242±0.006 n 100 2.200±0.002 d 2.069±0.004 k 3.035±0.004 k 0.940±0.004 l 1.379±0.02 m 3.026±0.002 m 150 3.060±0.001 k 1.797±0.002 d 2.776±.0.002 d 0.587±0.002 e 0.907±0.005h 2.433±0.004i 200 3.328±0.006 m 1.932±0.003 i 2.244±0.001 i 0.581±0.001 d 0.674±0.002 d 2.086±0.001 g Ascorbic acid 250 ppm 0 1.713±0.007 a 2.473±0.005 n 3.222±0.002 n 1.443±0.001 o 1.880±0.001 p 3.842±0.001 p 100 1.743±0.007 b 2.102±0.003 l 3.185±0.004 l 1.205±0.003 n 1.827±0.006 o 3.319±0.004 o 150 2.438±0.002 g 1.874±0.002 f 2.787±0.002 f 0.768±0.001 j 1.143±0.002 k 3.007±0.002 l 200 2.716±0.004 h 1.969±0.002 j 2.567±0.001 j 0.725±0.001 h 0.945±0.001 i 2.745±0.002 k Discussion Plants under salt stress have unique physiological and biochemical alterations that impact them from the beginning of the stress to maturity (Munns 2002 ). Significant decreases in growth metrics, such as root and shoot length, fresh and dry biomass, and leaf area, were seen in plants cultivated under different salt levels in this study. These consequences are linked to salt stress-induced cell shrinkage and dehydration, which limits cell elongation and division and, in turn, lowers the rate at which roots, shoots, and leaves grow. Osmotic alterations outside the roots are linked to these reactions, which hinder water intake and interfere with cell-water connections (Munns 2005 ). Our data support the conclusions of (Ghoulam et al. 2002), that demonstrated a discernible decrease in growth features due to salt, and with the investigation of (Kaya et al. 2018). Combining selenium, α-tocopherol, and ascorbic acid increased the growth of maize plants under salt stress. By strengthening antioxidant defences and regulating the metabolism of secondary metabolites, selenium in particular helps to regulate development (Jiang et al. 2017 ). Ascorbic acid helps plants grow and become more resilient to stress, in part by facilitating the production of proteins that are high in hydroxyproline (Alamri et al. 2018 ). Because chloroplasts are extremely sensitive to salinity, large salt concentrations can destabilise pigment–protein complexes and cause structural damage. Under salt stress, carotenoids suffer as well, and the amount of chlorophyll drops because of an increase in chlorophyllase activity (Mosavi et al. 2018 ). Changes in nitrogen metabolism, including increased synthesis of proline, which is essential for osmotic regulation, may be connected to the decrease in chlorophyll content. Glutamate kinase activity, the primary enzyme that starts proline production, is further stimulated by salt stress. (Shahbaz et al. 2022 ; And and Dawood 2014). Research by (El-Sawy 2009 and Hassanein et al. 2009 ) This suggests that decreased uptake of essential ions such as Mg²⁺ and Fe³⁺, which are essential for chlorophyll biosynthesis under stress, may be linked to the decline in plant pigment levels. Salinity also dramatically lowers stomatal conductance, photosynthetic electron transport efficiency, and carbon assimilation (Sun et al. 2016 ). The buildup of Na + and Cl + ions in plant tissues is a major sign of salt-induced stress, which upsets physiological equilibrium by changing the uptake of vital soil elements (Carvalho et al. 2020 ). In both stress and non-stress situations, enough selenium improves CO₂ assimilation and chlorophyll fluorescence, which increases photosynthetic efficiency (Alyemeni et al. 2018 ). Additionally, applying selenium to stressed plants promoted growth and raised the amount of photosynthetic pigment in Melissa officinalis L. (Habibi 2017 ), In contrast, Selenium raised K + and reduced Na + buildup in the roots and shoots. (Shekari, Abbasi, and Mustafavi 2017). A-tocopherol administered exogenously was discovered by (Das and Roychoudhury 2014) to significantly increase photosynthetic pigment concentrations. Plant leaves' photosynthetic membranes contain a-tocopherol (Grilo et al. 2014 ). Tocopherols are important for chloroplast heat dissipation, scavenge reactive oxygen species (ROS), and prevent lipid peroxidation. They are found in large quantities in cellular membranes (Falk and Munné-Bosch 2010). According to (Kruk and Strzalka 2001), In photosystem II, α-tocopherol protects chloroplasts by reducing cytochrome b 559 activity during cyclic electron flow. As mentioned by (Semida et al. 2016 ), Consistent with observations observed in other crops, exogenous α-tocopherol increased Antioxidant actions, both enzymatic and non-enzymatic, preserved cellular turgor, and enhanced photosynthetic efficiency in onion plants under salt stress. Solute loss, also known as electrolyte leakage, from plant cells can be measured to indirectly measure membrane damage (Ekmekçi et al. 2008), and assessing the membrane stability index (Ali and Abbas 2003). The findings unequivocally demonstrate that, in comparison to controls, high salinity considerably decreased the membrane stability index and increased electrolyte leakage in maize plants Jamil et al. ( 2012 ), It was shown that larger amounts of salt, in particular, resulted in more damage to cell membranes. (Hniličková et al. 2019) Higher salt concentrations were shown to enhance electrolyte leakage in all species. Leakage in Lactuca sativa reached 27.5%, indicating a significant difference between the control and 50 mmol/L NaCl treatments. Additionally, (Mahmoudi et al. 2011 ) With rising NaCl concentrations, electrolyte leakage in lettuce roots and leaves was seen to increase gradually. As a membrane-bound antioxidant and scavenger of reactive oxygen species (ROS), α-tocopherol assisted in lowering lipid peroxidation in maize plants (Orabi and Abdelhamid 2016). Exogenous α-tocopherol increased antioxidant enzyme activity and decreased MDA levels, which lessened the negative effects of seawater salinity on faba beans ( Semida et al. 2016 ). As previously reported, ascorbic acid application successfully decreased membrane lipid peroxidation in maize, most likely by strengthening the plant's antioxidant defence mechanism (Ahmad et al. 2016; Alyemeni et al. 2018 ). Supplementing with selenium has been shown to increase the uptake of nitrogen, potassium, and calcium, as well as amino acid synthesis and stress signalling, all of which help wheat become more resistant to salt (Elkelish et al. 2019 ). When selenium-amino acids were added to proteins, their biological activity changed, encouraging cucumber seedlings under salt stress to produce more proline (Hasanuzzaman et al. 2011). When α-tocopherol was applied exogenously to maize, it improved nitrogen metabolism, which led to increased proline buildup, better growth, and more yield components (Buschmann and Lichtenthaler 1979 ). Applying ascorbic acid to maize increased its protein content and nitrogen metabolism, according to (El-Bassiouny and Sadak 2015). Moreover, ascorbic acid and α-tocopherol applied topically significantly raised the amounts of protein and carbohydrates in flax cultivars under salt stress (Sadak and Dawood 2014). Carbohydrates, phenolics, and flavonoids are frequently linked to active osmotic adjustment, as noted by (Stepien and Klobus 2005 ; Li et al. 2010 ). The amount of total carbohydrates, phenolics, and flavonoids in maize plants decreased gradually as the salinity level increased under NaCl. Growth reductions were reflected in changes in carbohydrate content, which connected lower photosynthetic production to lower shoot dry weight. In salt-stressed maize, the decrease in chlorophyll and carbohydrates suggests either decreased photosynthesis or higher diversion of carbohydrates to other metabolic pathways (Handa et al. 1983 ; Shaddad et al. 1990 ). Abiotic and biotic stressors, such as salinity, often affect the synthesis of phenolics (Parida et al. 2004 ). Decreased activity of the essential enzyme that initiates the phenylpropanoid pathway, phenylalanine ammonia-lyase (PAL), which is necessary for plants to produce polyphenolic chemicals such flavonoids, phenylpropanoids, and lignin, is thought to be the cause of the drop in phenolic synthesis under stress (FRITZ et al. 1976; Tanaka et al. 1989 ). Furthermore, the absorption of potassium and phosphorus, which are necessary for secondary metabolites like polyphenols, is decreased at high salt concentrations (Rezazadeh et al. 2012 ). However, some research found that plants treated with salt had fewer phenolic chemicals (Wahid and Ghazanfar 2006; Blasco et al. 2013 ), others indicated enhanced phenolic biosynthesis (Burchard et al. 2000 ; Mahmoudi et al. 2010 ). The negative effects of salinity were partially mitigated by the individual administrations of Na₂SeO₄, α-tocopherol, and ascorbic acid, which usually improved the accumulation of carbohydrates, phenolics, and flavonoids in salt-stressed maize. Because of their absorption and storage in the green tissues, supplements containing sodium selenate or α-tocopherol under salt stress markedly increased physico-biochemical features, such as proteins, carbohydrates, carotenoids, and chlorophyll. In Melissa officinalis L. under Na + stress, selenium treatment also had a favourable impact on growth, photosynthetic pigments, and total carbohydrate content (Habibi 2017 ). α-tocopherol's antioxidant qualities were essential in reducing the negative effects of ROS brought on by salt stress (Attallah et al. 2013 ). By stabilising biological membranes and limiting lipid peroxidation, α-tocopherol in plastid membranes protected chloroplasts, the main location for sugar production in plants. (Wang and Quinn 2000; Sharma et al. 2012 ; Spicher, Glauser, and Kessler 2016). By increasing photosynthesis, boosting nutrient uptake, and decreasing the buildup of Na⁺ in tissues, exogenous ascorbic acid enabled maize plants treated with Na⁺ withstand oxidative damage. Likewise, under drought stress, seed priming with ascorbic acid raised Vicia faba's leaf chlorophyll, hydration status, and total carbohydrate content (Azooz et al. 2013 ). Ascorbic acid stimulated transpiration, photosynthesis, plant development, and the amount of carbohydrates (Naz et al. 2016; Alamri et al. 2018 ; Ibrahim et al. 2019 ) showed that when exposed to 5000 ppm salinity, the phenolic and flavonoid levels rose and interacted with 150 ppm ascorbic acid. (Wu et al. 2010 ), after being treated with ascorbic acid, Geranium sibiricum L. also showed an increase in total phenolic content. In addition to increased phenylalanine ammonia-lyase (PAL) and catalase (CAT) activities during the study period under constant NaCl levels, salt stress enhances the activities of amylases, proteases, polyphenol oxidase, and peroxidase. Salinity has been shown to increase amylase and protease activities in the cotyledons of sprouting cotton seeds, but it also delays reserve mobilisation, which is linked to decreased activity of reserve-degrading enzymes. (Ashraf et al. 2002 ), whereas during late seedling establishment it reduces the activities of lipases, amylases, and proteases in cashew cotyledons (Marques et al. 2013). (Zeid et al. 2019 ), Optimal quantities of ascorbic acid, gibberellic acid, sodium selenate (Na₂SeO₄), or selenium nanoparticles (SeNPs) enhanced the activities of α-amylase, β-amylase, and protease in seedlings, but higher concentrations resulted in a decrease. When α-tocopherol was applied to maize under water stress, it reduced amylase activity and encouraged the buildup of carbohydrates (Terzi et al. 2018). (Abdallah et al. 2013 ) The activities of α-amylase, β-amylase, and protease in seedlings were increased by optimal amounts of ascorbic acid, gibberellic acid, sodium selenate (Na₂SeO₄), or selenium nanoparticles (SeNPs); however, higher concentrations caused a decrease. α-tocopherol decreased amylase activity and promoted the accumulation of carbohydrates in maize under water stress (Hammerschmidt et al. 1982; Fukasawa-Akada et al. 1996 ; Thipyapong and Steffens 1997). In order to reduce the excessive formation of reactive oxygen species (ROS) brought on by salt, selenium is essential for both enzymatic processes like catalase (CAT) and peroxidase (POD) activities as well as non-enzymatic mechanisms involving phytochelatins and glutathione (Zong et al. 2009 ). It has been said that applying Se is crucial for boosting ROS scavenging activity and for naturally boosting plant antioxidant activity (Shalaby et al. 2017 ). Additionally, it has been demonstrated that supplementing with Se reduces the production of H 2 O 2 during salt stress, as seen by improvements in the antioxidant enzyme activities (POD and CAT) in canola and dill seedlings (Hasanuzzaman et al. 2011; Shekari et al. 2017). Exogenous α-tocopherol's beneficial effects on a range of physio-biochemical characteristics most likely result from its involvement in several physiological functions and its internal translocation inside the plant, which improves overall stress tolerance (Kumar et al. 2013 ). As a cofactor for many antioxidant enzymes, ascorbic acid supports a number of physiological functions, such as controlling the amount of proteins and nucleic acids, membrane permeability, photosynthesis, root ion uptake, respiration, seed germination, and enzyme activity in a variety of plant species (Tavili et al. 2009 ; Ozdener and Kutbay 2008; Ejaz et al. 2012). (Farooq et al. 2020 ) showed that CAT and POD enzyme activities rose in response to water stress, which is in line with previous findings of increased PPO, CAT, and POD enzyme activities in wheat treated with ascorbic acid (Shafiq et al. 2014 ; Hafez and Gharib 2016 ). Protein profiles change as a result of salinity stress, and stress-specific proteins that shield cells from salt damage are also induced. In control plants, vitamins have been demonstrated to alter gene expression, increasing the synthesis of preexisting proteins and encouraging the emergence of new protein bands (Bassuony et al. 2008 ; Al-Hafedh et al. 2008). Vitamins have a substantial impact on the amount and quality of protein composition, which may improve growth and productivity, as evidenced by the observed increase in the intensity of original protein bands in control plants. Furthermore, one important defence against salt stress is the de novo creation of particular proteins (Hassanein et al. 2009 ; Eid 2019 ). Many biosynthetic and signalling pathways control the genetically complicated trait of salt tolerance in plants. Numerous investigations have documented changes in polypeptide patterns in response to salt stress, both qualitatively and quantitatively (Chawla et al. 2010 ; Arefian et al. 2019 ; Baniulis et al. 2020 ). Salinity decreased the protein content of pigeon peas (Cajanus cajan), with the decline being more noticeable at 100 mM NaCl than at 50 mM NaCl (Chavan et al. 2018 ). Through alterations in one-dimensional SDS-PAGE profiles, the current work shows that gene expression in maize is modulated by both NaCl and individual administrations of sodium selenate, α-tocopherol, and ascorbic acid. The seeds treated with α-tocopherol produced two bands at 59.296 and 39.285 kDa, ascorbic acid produced bands at 80.472 and 67.439 kDa, and presoaking in sodium selenate produced a novel protein band at 82.846 kDa. These recently produced proteins most likely serve as osmoprotectants, metabolic enzymes, ROS scavengers, signal transducers, or are involved in the biogenesis, degradation, and defence processes of cells (Sairam and Tyagi 2004; (Chattopadhyay et al. 2011 ; Lisa et al. 2011 ; Kappel et al. 2020 ). While SDS-PAGE cannot definitively identify proteins, the observed patterns indicate that selenium, α-tocopherol, and ascorbic acid modulate the expression of constitutive and stress-inducible proteins. Bands in the 95–72 kDa range may correspond to Rubisco or other photosynthetic proteins, 67–59 kDa to stress-related enzymes, and lower molecular weight bands (44–34 kDa) to regulatory or defense proteins, highlighting the physiological adjustments of maize under abiotic stress and antioxidant treatments. Excessive Na + buildup in plants interferes with stomatal control by upsetting K + equilibrium. Therefore, maintaining a high K⁺/Na⁺ ratio is crucial for salt tolerance (Shabala et al. 2005 ; Abbasi et al. 2015 ). Crops adjust to salinity by limiting the movement of Na + and Cl + ions to stems and leaf sheaths, which reduces their buildup in more vulnerable leaf blades (Isla and Aragüés 2010). Sustaining K + intake, decreasing K + efflux, restricting Na + influx, and encouraging Na + efflux from cells are all necessary to maintain the cytosolic K + /Na + ratio (Wakeel et al. 2011 ). It has been demonstrated that high concentrations of NaCl in the growth medium decrease K + ion absorption (Faheed 2012 ; Noreen et al. 2017 ). Relative water content is decreased when hydraulic conductivity is decreased due to salinity-induced Na + buildup in roots. On the other hand, supplementing with selenium (Na₂SeO₄) reduces Na⁺ toxicity, encourages root growth, and improves water transfer to shoots, all of which boost overall plant growth (Rietz and Haynes 2003 ). Because proline production is necessary for preserving protein stability, saline soils interfere with nitrogen uptake, accumulation, and metabolism (Carillo et al. 2005 ; Szabados and Savoure 2010). It has been demonstrated that selenium enhances nutrient uptake and translocation in a variety of crops, boosting yield and growth (Shahzadi et al. 2017 ). Interestingly, when compared to untreated plants, selenite-treated plants showed greater K + /Na + ratios and lower Na + concentrations (Subramanyam et al., 2019). Applying selenium has been associated with a decrease in Na + buildup, which raises the K + /Na + ratio in plants that produce garlic (Allium sativum L.) and dill (Anethum graveolens) (Shekari, Abbasi, and Mustafavi 2017); (Astaneh et al. 2018 ). In a similar vein, applying selenium to Melissa officinalis L. improved growth, photosynthetic pigments, and total amino acid content while decreasing Na + and increasing K + levels in Anethum graveolens plants (Habibi 2017 ;Shekari et al. 2017). Antioxidants like α-tocopherol, which improve osmotolerance and control processes like membrane permeability and nutrient uptake from the soil, are responsible for the increased nutritional content (Orabi and Abdelhamid 2016). Applying α-tocopherol has been associated with increased leaf ion content, which improves nutrient uptake and has a beneficial effect on plant growth (Buschmann and Lichtenthaler 1979 ); (Sadak and Dawood 2014). Additionally, α-tocopherol has been shown to be successful in reducing the harmful effects of saltwater salinity on faba beans (Orabi and Abdelhamid 2016). Applying α-tocopherol to flax cultivars has been demonstrated to improve plant growth and yield by raising Ca + 2 , Mg + 2 , and K + levels while decreasing Na + and Cl − levels (Sadak et al. 2020 ; Semida et al. 2014 ). While K⁺ content in both tissues dropped as NaCl concentration increased, ascorbic acid treatment dramatically inhibited Na⁺ buildup in faba bean roots and shoots (Loutfy et al. 2019 ). Additionally, under salt stress conditions, ascorbic acid has been shown to increase the levels of K + , Ca + 2 , and Mg + 2 while decreasing the buildup of Na + (El-Bassiouny and Sadak 2015). Conclusions According to the present study, salt-induced oxidative stress markedly inhibits plant growth and development by impairing photosynthetic efficiency, disrupting nutrient homeostasis, and elevating multiple stress biomarkers in a concentration-dependent manner. In contrast, the exogenous application of selenium, α-tocopherol, and ascorbic acid significantly improves growth performance in salt-stressed plants. This protective treatment enhances chlorophyll biosynthesis and stimulates the accumulation of secondary metabolites. Notably, ascorbic acid proved to be more effective than selenium and α-tocopherol in alleviating salt stress. Its superior efficacy is attributed to efficient scavenging of reactive oxygen species (ROS), enhanced osmolyte (proline) accumulation, and reinforcement of the antioxidant defense system. Consequently, ascorbic acid improves plant water relations, growth attributes, and photosynthetic efficiency under saline conditions. Furthermore, this treatment induces the expression of proteins associated with salt-stress tolerance. Collectively, these findings highlight a promising, eco-friendly, and cost-effective strategy for mitigating salt toxicity in agricultural systems worldwide. Declarations Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding This research was funded by Prince Sattam bin Abdulaziz University under project number PSAU/2025/01/32873 Author Contribution RK and MY– Conceive and conceptualize the idea; FB and SH– Investigated and collected data; RK, MY, AG – Analyzed data and prepared graphs; MY and RK– Prepared original draft and revised, MY– Edited and content improvement, MR, HN, MA, MR, YD – Statistical analysis and revised the draft. Acknowledgments The authors would like to thank the Botany and microbiology department, Faculty of Science, Benha University. For permitting us to carry out the experiment using their laboratory facilities. Also, the authors extend their appreciation to prince Sattam bin Abdulaziz University for funding this research work through the project number (PSAU/2025/01/32873). Availability of data and material (data transparency) The data that support the findings of this study are available from Prince Sattam bin Abdulaziz University and Benha University but restrictions apply to the availability of these data, which were used under license for the current study, and so are not publicly available. Data are however available from the authors upon reasonable request and with permission of Prince Sattam bin Abdulaziz University and Benha University. References Abbasi, Ghulam Hasan, Javaid Akhtar, Rafiq Ahmad, Moazzam Jamil, Muhammad Anwar-ul-Haq, Shafaqat Ali, and Muhammad Ijaz. 2015. “Potassium Application Mitigates Salt Stress Differentially at Different Growth Stages in Tolerant and Sensitive Maize Hybrids.” Plant Growth Regulation 76 (1): 111–25. Abdallah, M M, Abd El-Monem AA, R A Hassanein, and H M S El-Bassiouny. 2013. “Response of Sunflower Plant to the Application of Certain Vitamins and Arbuscular Mycorrhiza under Different Water Regimes.” Aust J Basic Appl Sci 7: 915–32. Aebi, Hugo. 1984. “[13] Catalase in Vitro.” In Methods in Enzymology , 105:121–26. Elsevier. Ahmad, Parvaiz, E F Abd Allah, Abeer Hashem, Maryam Sarwat, and Salih Gucel. 2016. “Exogenous Application of Selenium Mitigates Cadmium Toxicity in Brassica Juncea L.(Czern & Cross) by up-Regulating Antioxidative System and Secondary Metabolites.” Journal of Plant Growth Regulation 35 (4): 936–50. Al-Hafedh, Yousef S, Aftab Alam, and Mohamed Salaheldin Beltagi. 2008. “Food Production and Water Conservation in a Recirculating Aquaponic System in Saudi Arabia at Different Ratios of Fish Feed to Plants.” Journal of the World Aquaculture Society 39 (4): 510–20. Alamri, Saud A, Manzer H Siddiqui, Mutahhar Y Y Al-khaishany, M Nasir, Hayssam M Ali, Ibrahim A Alaraidh, Abdulaziz A Alsahli, Hala Al-, and Mohammed Mateen. 2018. “Ascorbic Acid Improves the Tolerance of Wheat Plants to Lead Toxicity.” Journal of Plant Interactions 9145 (1): 409–19. https://doi.org/10.1080/17429145.2018.1491067 . Ali, Refaat Mohamed, and H. M. Abbas. 2003. “Response of Salt Stressed Barley Seedlings to Phenylurea.” Plant, Soil and Environment 49 (4): 158–62. Alyemeni, Mohammed Nasser, Mohammad Abass Ahanger, Leonard Wijaya, Pravej Alam, Renu Bhardwaj, and Parvaiz Ahmad. 2018. “Selenium Mitigates Cadmium-Induced Oxidative Stress in Tomato (Solanum Lycopersicum L.) Plants by Modulating Chlorophyll Fluorescence, Osmolyte Accumulation, and Antioxidant System.” Protoplasma 255 (2): 459–69. And, Sadak, and Dawood. 2014. “Role of Ascorbic Acid and $ α $ Tocopherol in Alleviating Salinity Stress on Flax Plant (Linum Usitatissimum L.).” Journal of Stress Physiology & Biochemistry 10 (1): 93–111. Arefian, Mohammad, Saeedreza Vessal, Saeid Malekzadeh-Shafaroudi, Kadambot H M Siddique, and Abdolreza Bagheri. 2019. “Comparative Proteomics and Gene Expression Analyses Revealed Responsive Proteins and Mechanisms for Salt Tolerance in Chickpea Genotypes.” BMC Plant Biology 19 (1): 300. https://doi.org/10.1186/s12870-019-1793-z . Arif, Yamshi, Priyanka Singh, Husna Siddiqui, Andrzej Bajguz, and Shamsul Hayat. 2020. “Salinity Induced Physiological and Biochemical Changes in Plants: An Omic Approach towards Salt Stress Tolerance.” Plant Physiology and Biochemistry 156: 64–77. Ashraf, M Y, G Sarwar, M Ashraf, R Afaf, and A Sattar. 2002. “Salinity Induced Changes in $ α $ -Amylase Activity during Germination and Early Cotton Seedling Growth.” Biologia Plantarum 45 (4): 589–91. Astaneh, Rozita Khademi, Sahebali Bolandnazar, Fariborz Zaare Nahandi, and Shahin Oustan. 2018. “The Effects of Selenium on Some Physiological Traits and K, Na Concentration of Garlic (Allium Sativum L.) under NaCl Stress.” Information Processing in Agriculture 5 (1): 156–61. Attallah, Abdelfattah M, Faisal A Bughdadi, Atef M El-Shazly, and Hisham Ismail. 2013. “Immunodetection of Fasciola Gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis.” Clinical and Vaccine Immunology 20 (10): 1569–77. Azooz, Mohamed M, Abdullah M Alzahrani, Magdy M Youssef, and others. 2013. “The Potential Role of Seed Priming with Ascorbic Acid and Nicotinamide and Their Interactions to Enhance Salt Tolerance in Broad Bean ('Vicia Faba’L.).” Australian Journal of Crop Science 7 (13): 2091–2100. Baniulis, Danas, Monika Sirgediene, Perttu Haimi, Inga Tamošiune, and Darius Danusevičius. 2020. “Constitutive and Cold Acclimation-Regulated Protein Expression Profiles of Scots Pine Seedlings Reveal Potential for Adaptive Capacity of Geographically Distant Populations.” Forests 11 (1): 89. Bassuony, F M, R A Hassanein, D M Baraka, and R R Khalil. 2008. “Physiological Effects of Nicotinamide and Ascorbic Acid on Zea Mays Plant Grown under Salinity Stress. II-Changes in Nitrogen Constituents, Protein Profiles, Protease Enzyme and Certain Inorganic Cations.” Australian Journal of Basic and Applied Sciences 2 (3): 350–59. Bates, L S, R P Waldren, and I D Teare. 1973. “Rapid Determination of Free Proline for Water-Stress Studies.” Plant and Soil 39 (1): 205–7. Blasco, Begon?a, Rocio Leyva, Luis Romero, and Juan Manuel Ruiz. 2013. “Iodine Effects on Phenolic Metabolism in Lettuce Plants under Salt Stress.” Journal of Agricultural and Food Chemistry 61 (11): 2591–96. Burchard, P, W Bilger, and G Weissenböck. 2000. “Contribution of Hydroxycinnamates and Flavonoids to Epidermal Shielding of UV-A and UV-B Radiation in Developing Rye Primary Leaves as Assessed by Ultraviolet-Induced Chlorophyll Fluorescence Measurements.” Plant, Cell & Environment 23 (12): 1373–80. Buschmann, C, and H K Lichtenthaler. 1979. “The Influence of Phytohormones on Prenyl Lipid Composition and Photosynthetic Activities of Thylakoids.” Advances in Biochemistry and Physiology of Plant Lipids. Elsevier, Amserdam , 145–50. Carillo, Petronia, Gabriella Mastrolonardo, Francesco Nacca, and Amodio Fuggi. 2005. “Nitrate Reductase in Durum Wheat Seedlings as Affected by Nitrate Nutrition and Salinity.” Functional Plant Biology 32 (3): 209–19. Carvalho, Marcia E.A., Paulo R.C. Castro, Marcin Kozak, and Ricardo A. Azevedo. 2020. “The Sweet Side of Misbalanced Nutrients in Cadmium-Stressed Plants.” Annals of Applied Biology 176 (3): 275–84. https://doi.org/10.1111/aab.12569 . Chattopadhyay, Arnab, Pratigya Subba, Aarti Pandey, Deepti Bhushan, Rajiv Kumar, Asis Datta, Subhra Chakraborty, and Niranjan Chakraborty. 2011. “Analysis of the Grasspea Proteome and Identification of Stress-Responsive Proteins upon Exposure to High Salinity, Low Temperature, and Abscisic Acid Treatment.” Phytochemistry 72 (10): 1293–1307. Chavan, Nilesh S, Prashant B Kardile, and Roshan A Diware. 2018. “Effect of Salt Stress on Protein Profile of Different Varieties of Cajanus Cajan Using SDS Page.” Journal of Pharmacognosy and Phytochemistry 7 (6): 1258–62. Chawla, S, S C Goyal, K S Datta, R Angrish, and others. 2010. “Protein Profile Changes under Salt-Boron Toxicity and Its Regulation by Hydrogen Peroxide and Glutathione in Pigeonpea (Cajanus Cajan L.).” Indian Journal of Plant Physiology 15 (4): 378–83. Clark, J M, and R L Switzer. 1977. “In Experimental Biochemistry 2nd Ed.W.H. Freeman &Company, San Francisco.” Article. Das, Kaushik, and Aryadeep Roychoudhury. 2014. “Reactive Oxygen Species (ROS) and Response of Antioxidants as ROS-Scavengers during Environmental Stress in Plants.” Frontiers in Environmental Science 2: 53. Eid, Manal Hassan. 2019. “Gene Expression and Molecular Differences in Two Wheat Genotypes under Salt and Drought Stresses.” Egyptian Journal of Agronomy 41 (2): 93–104. Ejaz, Batool, Zahoor Ahmad Sajid, and Faheem Aftab. 2012. “Effect of Exogenous Application of Ascorbic Acid on Antioxidant Enzyme Activities, Proline Contents, and Growth Parameters of Saccharum Spp. Hybrid Cv. HSF-240 under Salt Stress.” Turkish Journal of Biology 36 (6): 630–40. Ekmekçi, Yasemin, Deniz Tanyolac, and Beycan Ayhan. 2008. “Effects of Cadmium on Antioxidant Enzyme and Photosynthetic Activities in Leaves of Two Maize Cultivars.” Journal of Plant Physiology 165 (6): 600–611. El-Bassiouny, Hala M.S., and Mervat Sh Sadak. 2015. “Impact of Foliar Application of Ascorbic Acid and Alpha-Tocopherol on Antioxidant Activity and Some Biochemical Aspects of Flax Cultivars under Salinity Stress.” Acta Biologica Colombiana 20 (2): 209–22. https://doi.org/10.15446/abc.v20n2.43868 . El-Sawy, O E H. 2009. “Protective Effects of Polyamines on Wheat Plants Irrigated by Seawater.” Ph. D. Thesis, Fac. Sci., Mans. Univ., Egypt. Elkelish, Amr A, Mona H Soliman, Haifa A Alhaithloul, and Mohamed A El-Esawi. 2019. “Selenium Protects Wheat Seedlings against Salt Stress-Mediated Oxidative Damage by up-Regulating Antioxidants and Osmolytes Metabolism.” Plant Physiology and Biochemistry 137: 144–53. Eltayeb, Amin Elsadig, Naoyoshi Kawano, Ghazi Hamid Badawi, Hironori Kaminaka, Takeshi Sanekata, Toshiyuki Shibahara, Shinobu Inanaga, and Kiyoshi Tanaka. 2007. “Overexpression of Monodehydroascorbate Reductase in Transgenic Tobacco Confers Enhanced Tolerance to Ozone, Salt and Polyethylene Glycol Stresses.” Planta 225 (5): 1255–64. Fadeel, A A. 1962. “Location and Properties of Chloroplasts and Pigment Determination in Roots.” Physiologia Plantarum 15 (1): 130–46. Faheed, Fayza A. 2012. “Comparative Effects of Four Herbicides on Physiological Aspects in Triticum Sativum L.” African Journal of Ecology 50 (1): 29–42. Falk, Jon, and Sergi Munné-Bosch. 2010. “Tocochromanol Functions in Plants: Antioxidation and Beyond.” Journal of Experimental Botany 61 (6): 1549–66. Farooq, Ayesha, Shazia Anwer Bukhari, Nudrat A Akram, Muhammad Ashraf, Leonard Wijaya, Mohammed Nasser Alyemeni, and Parvaiz Ahmad. 2020. “Exogenously Applied Ascorbic Acid-Mediated Changes in Osmoprotection and Oxidative Defense System Enhanced Water Stress Tolerance in Different Cultivars of Safflower (Carthamus Tinctorious L.).” Plants 9 (1): 104. FRITZ, Taylor, Tatsuma ITO, Malek JAZIRI, John-Patrick SMITH, Alejandro GONZALEZ, Austin KRAJICEK, and Yoshihito NISHIOKA. 1976. “8 6 JPN.” Fukasawa-Akada, Tomoko, Shain-dow Kung, and John C Watson. 1996. “Phenylalanine Ammonia-Lyase Gene Structure, Expression, and Evolution in Nicotiana.” Plant Molecular Biology 30 (4): 711–22. Ghoulam, Cherki, Ahmed Foursy, and Khalid Fares. 2002. “Effects of Salt Stress on Growth, Inorganic Ions and Proline Accumulation in Relation to Osmotic Adjustment in Five Sugar Beet Cultivars.” Environmental and Experimental Botany 47 (1): 39–50. Gong, D H, G Z Wang, W T Si, Y Zhou, Z Liu, and J Jia. 2018. “Effects of Salt Stress on Photosynthetic Pigments and Activity of Ribulose-1, 5-Bisphosphate Carboxylase/Oxygenase in Kalidium Foliatum.” Russian Journal of Plant Physiology 65 (1): 98–103. Grilo, Evellyn Câmara, Priscila Nunes Costa, Cristiane Santos Sânzio Gurgel, Andressa Fernanda de Lima Beserra, Fernanda Niéce de Souza Almeida, and Roberto Dimenstein. 2014. “Alpha-Tocopherol and Gamma-Tocopherol Concentration in Vegetable Oils.” Food Science and Technology 34 (2): 379–85. Habibi, G. 2017. “Selenium Ameliorates Salinity Stress in Petroselinum Crispum by Modulation of Photosynthesis and by Reducing Shoot Na Accumulation.” Russian Journal of Plant Physiology 64 (3): 368–74. Hafez, E M, and H S Gharib. 2016. “Effect of Exogenous Application of Ascorbic Acid on Physiological and Biochemical Characteristics of Wheat under Water Stress.” Int. J. Plant Prod 10 (4): 579–96. Hammerschmidt, R, E M Nuckles, and J Kuć. 1982. “Association of Enhanced Peroxidase Activity with Induced Systemic Resistance of Cucumber to Colletotrichum Lagenarium.” Physiological Plant Pathology 20 (1): 73–82. Handa, Sangita, Ray A Bressan, Avtar K Handa, Nicholas C Carpita, and Paul M Hasegawa. 1983. “Solutes Contributing to Osmotic Adjustment in Cultured Plant Cells Adapted to Water Stress.” Plant Physiology 73 (3): 834–43. Harris, D, A Rashid, G Miraj, M Arif, and H Shah. 2007. “On-Farmseed Priming with Zinc Sulphate SolutionA Cost-Effective Way to Increase the Maize Yields of Resource-Poor Farmers.” Field Crops Research 102 (2): 119–27. Harris, David, Abdul Rashid, Ghazal Miraj, Mohammed Arif, and Mohammed Yunas. 2008. “On-Farmseed Priming with Zinc in Chickpea and Wheat in Pakistan.” Plant and Soil 306 (1–2): 3–10. Hartree, E Fo. 1972. “Determination of Protein: A Modification of the Lowry Method That Gives a Linear Photometric Response.” Article. Analytical Biochemistry 48 (2): 422–27. Hasanuzzaman, Mirza, Mohammad Anwar Hossain, and Masayuki Fujita. 2011. “Selenium-Induced up-Regulation of the Antioxidant Defense and Methylglyoxal Detoxification System Reduces Salinity-Induced Damage in Rapeseed Seedlings.” Biological Trace Element Research 143 (3): 1704–21. Hassanein, R A, F M Bassuony, D M Baraka, and R R Khalil. 2009. “Physiological Effects of Nicotinamide and Ascrobic Acid on Zea Mays Plant Grown Under Salinity Stress. I-Changes in Growth, Some Relevant Metabolic Activities and Oxidative Defense Systems .” Journal of Agriculture and Biological Sciences 5 (1): 72–81. Havir, Evelyn A, and Neil A McHale. 1987. “Biochemical and Developmental Characterization of Multiple Forms of Catalase in Tobacco Leaves.” Plant Physiology 84 (2): 450–55. Hniličková, Helena, František Hnilička, Matyáš Orsák, and Václav Hejnák. 2019. “Effect of Salt Stress on Growth, Electrolyte Leakage, Na + and K+ Content in Selected Plant Species.” Plant, Soil and Environment 65 (2): 90–96. Hodges, D Mark, John M DeLong, Charles F Forney, and Robert K Prange. 1999. “Improving the Thiobarbituric Acid-Reactive-Substances Assay for Estimating Lipid Peroxidation in Plant Tissues Containing Anthocyanin and Other Interfering Compounds.” Planta 207 (4): 604–11. Ibrahim, Gamil E, Sami A Metwally, H Bedour, and M Sharbat. 2019. “Effect of Salinity and Ascorbic Acid on Geranium Volatile and Antioxidant Activity.” Plant Archives 19 (2): 750–57. Isla, Ramon, and Rámon Aragüés. 2010. “Yield and Plant Ion Concentrations in Maize (Zea Mays L.) Subject to Diurnal and Nocturnal Saline Sprinkler Irrigations.” Field Crops Research 116 (1–2): 175–83. Jamil, Muhammad, Muhammad Ashraf, S Rehman, Mushtaq Ahmad, and Eui Shik Rha. 2012. “Salinity Induced Changes in Cell Membrane Stability, Protein and RNA Contents.” African Journal of Biotechnology 11 (24): 6476–83. Jiang, Chaoqiang, Chaolong Zu, Dianjun Lu, Qingsong Zheng, Jia Shen, and Huoyan Wang. 2017. “Effect of Exogenous Selenium Supply on Photosynthesis, Na + Accumulation and Antioxidative Capacity of Maize (Zea Mays L .) under Salinity Stress.” Nature Publishing Group 7 (February): 1–14. https://doi.org/10.1038/srep42039 . Kappel, Lisa, Martin Münsterkötter, György Sipos, Carolina Escobar Rodriguez, and Sabine Gruber. 2020. “Chitin and Chitosan Remodeling Defines Vegetative Development and Trichoderma Biocontrol.” PLoS Pathogens 16 (2): e1008320. Kar, Manoranjan, and Dinabandhu Mishra. 1976. “Catalase, Peroxidase, and Polyphenoloxidase Activities during Rice Leaf Senescence.” Plant Physiology 57 (2): 315–19. Kaya, Cengiz, Muhammad Ashraf, and Osman Sonmez. 2018. “Combination of Nitric Oxide and Thiamin Regulates Oxidative Defense Machinery and Key Physiological Parameters in Salt-Stressed Plants of Two Maize Cultivars Differing in Salinity Tolerance.” Advances in Agricultural Science 6 (1): 34–44. http://aaasjournal.org/submission/index.php/aaas/article/view/25 . Khalil, Radwan, Samia Haroun, Fardous Bassyoini, Aziza Nagah, and Mohammad Yusuf. 2021. “Salicylic Acid in Combination with Kinetin or Calcium Ameliorates Heavy Metal Stress in Phaseolus Vulgaris Plant.” Journal of Agriculture and Food Research 5: 100182. Klapheck, Sigrid, Ina Zimmer, and Heinz Cosse. 1990. “Scavenging of Hydrogen Peroxide in the Endosperm of Ricinus Communis by Ascorbate Peroxidase.” Plant and Cell Physiology 31 (7): 1005–13. Kruk, Jerzy, and Kazimierz Strzalka. 2001. “Redox Changes of Cytochrome b 559 in the Presence of Plastoquinones.” Journal of Biological Chemistry 276 (1): 86–91. Kumar, Sanjeev, R Singh, and Harsh Nayyar. 2013. “{ $ }\alpha{\ $ }-Tocopherol Application Modulates the Response of Wheat (Triticum Aestivum L.) Seedlings to Elevated Temperatures by Mitigation of Stress Injury and Enhancement of Antioxidants.” Journal of Plant Growth Regulation 32 (2): 307–14. Laemmli, Ulrich K. 1970. “Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4.” Nature 227 (5259): 680–85. Li, Yin, Zinan Wang, Xiaofen Sun, and Kexuan Tang. 2008. “Current Opinions on the Functions of Tocopherol Based on the Genetic Manipulation of Tocopherol Biosynthesis in Plants.” Journal of Integrative Plant Biology 50 (9): 1057–69. Li, Zhao Hui, Qiang Wang, Xiao Ruan, Cun De Pan, and De An Jiang. 2010. “Phenolics and Plant Allelopathy.” Molecules 15 (12): 8933–52. https://doi.org/10.3390/molecules15128933 . Lisa, Laisa A, Sabrina M Elias, M Sazzadur Rahman, Saima Shahid, Tetsushi Iwasaki, A K M Mahbub Hasan, Keiko Kosuge, Yasuo Fukami, and Zeba I Seraj. 2011. “Physiology and Gene Expression of the Rice Landrace Horkuch under Salt Stress.” Functional Plant Biology 38 (4): 282–92. Loutfy, Naglaa, Mohamed M Azooz, Ahmed M Hassanein, and Abdelkader Bassiony. 2019. “Potassium Synergize the Positive Effect of Ascorbic Acid on Some Morpho-Physiological Parameters of Salt Stressed Faba Bean Cultivars.” Egyptian Journal of Botany 59 (3): 735–51. Lowe, L E. 1993. “Water-Soluble Phenolic Materials.” Soil Sampling and Methods of Analysis. Canadian Society of Soil Science. CRC Press, Boca Raton, FL, USA , 409–12. Mahmoudi, Hela, Jun Huang, Margaret Y Gruber, Rym Kaddour, Mokhtar Lachaal, Zeineb Ouerghi, and Abdelali Hannoufa. 2010. “The Impact of Genotype and Salinity on Physiological Function, Secondary Metabolite Accumulation, and Antioxidative Responses in Lettuce.” Journal of Agricultural and Food Chemistry 58 (8): 5122–30. Mahmoudi, Hela, Rym Kaddour, Jun Huang, Nawel Nasri, Baâtour Olfa, Sabah MRah, Abdelali Hannoufa, Mokhtar Lachaâl, and Zeineb Ouerghi. 2011. “Varied Tolerance to NaCl Salinity Is Related to Biochemical Changes in Two Contrasting Lettuce Genotypes.” Acta Physiologiae Plantarum 33 (5): 1613–22. Malick, C P, and M B Singh. 1980. “Plant Enzymology and Histo Enzymology Kalyari’’Publishers New Delhi Pp, 286.” Marques, Elton Camelo, Paulo André Ferreira de Freitas, Nara L\’\idia Mendes Alencar, José Tarquinio Prisco, and Enéas Gomes-Filho. 2013. “Increased Na + and Cl- Accumulation Induced by NaCl Salinity Inhibits Cotyledonary Reserve Mobilization and Alters the Source-Sink Relationship in Establishing Dwarf Cashew Seedlings.” Acta Physiologiae Plantarum 35 (7): 2171–82. Mosavi, Nosrat, Mostafa Ebadi, Mahdi Khorshidi, and Nahid Masoudian. 2018. “Study of Some Physiological Characteristics of Potato Tissue under Salinity Stress.” International Journal of Farming and Allied Sciences 7 (1): 1–5. Munné-Bosch, Sergi. 2005. “The Role of $ α $ -Tocopherol in Plant Stress Tolerance.” Journal of Plant Physiology 162 (7): 743–48. ---. 2007. “{ $ }\alpha{\ $ }-Tocopherol: A Multifaceted Molecule in Plants.” Vitamins {&} Hormones 76: 375–92. Munné-Bosch, Sergi, and Leonor Alegre. 2002. “The Function of Tocopherols and Tocotrienols in Plants.” Critical Reviews in Plant Sciences 21 (1): 31–57. Munns, Rana. 2002. “Comparative Physiology of Salt and Water Stress.” Plant, Cell & Environment 25 (2): 239–50. ---. 2005. “Genes and Salt Tolerance: Bringing Them Together.” New Phytologist 167 (3): 645–63. Naz, HIRA, Nudrat Aisha Akram, and Muhammad Ashraf. 2016. “Impact of Ascorbic Acid on Growth and Some Physiological Attributes of Cucumber (Cucumis Sativus) Plants under Water-Deficit Conditions.” Pak J Bot 48 (3): 877–83. Nguyen, Thi Bich Thuy, Saichol Ketsa, and Wouter G van Doorn. 2003. “Relationship between Browning and the Activities of Polyphenoloxidase and Phenylalanine Ammonia Lyase in Banana Peel during Low Temperature Storage.” Postharvest Biology and Technology 30 (2): 187–93. Noreen, Sibgha, Ayesha Siddiq, Kousar Hussain, Shakeel Ahmad, and Mirza Hasanuzzaman. 2017. “Foliar Application of Salicylic Acid with Salinity Stress on Physiological and Biochemical Attributes of Sunflower (Helianthus Annuus L.) Crop.” Acta Scientiarum Polonorum-Hortorum Cultus 16 (2): 57–74. Orabi, Salwa A, and Magdi T Abdelhamid. 2016. “Protective Role of { $ }\alpha{\ $ }-Tocopherol on Two Vicia Faba Cultivars against Seawater-Induced Lipid Peroxidation by Enhancing Capacity of Anti-Oxidative System.” Journal of the Saudi Society of Agricultural Sciences 15 (2): 145–54. Ozdener, Yasemin, and H Guray Kutbay. 2008. “Effect of Salinity and Temperature on the Germination of Spergularia Marina Seeds and Ameliorating Effect of Ascorbic and Salicylic Acids.” Journal of Environmental Biology 29 (6): 959–64. Parida, Asish Kumar, Anath Bandhu Das, Yukika Sanada, and Prasanna Mohanty. 2004. “Effects of Salinity on Biochemical Components of the Mangrove, Aegiceras Corniculatum.” Aquatic Botany 80 (2): 77–87. Rautenkranz, Andreas A F, Liantje Li, Felix Machler, Enrico Martinoia, and Johann J Oertli. 1994. “Transport of Ascorbic and Dehydroascorbic Acids across Protoplast and Vacuole Membranes Isolated from Barley (Hordeum Vulgare L. Cv Gerbel) Leaves.” Plant Physiology 106 (1): 187–93. Rezazadeh, Amir, Azim Ghasemnezhad, Mojtaba Barani, and Telmah Telmadarrehei. 2012. “Effect of Salinity on Phenolic Composition and Antioxidant Activity of Artichoke (Cynara Scolymus L.) Leaves.” Res J Med Plant 6: 245–52. Rietz, D N, and R J Haynes. 2003. “Effects of Irrigation-Induced Salinity and Sodicity on Soil Microbial Activity.” Soil Biology and Biochemistry 35 (6): 845–54. Rostami, Mahdi, and Hossein Abbaspour. 2019. “EFFECT OF SELENIUM ON GROWTH AND PHYSIOLOGICAL TRAITS OF BASIL PLANT (Ocimum Basilicum L.) UNDER ARSENIC STRESS CONDITIONS.” REVISTA DE AGRICULTURA NEOTROPICAL 6 (3): 30–37. https://doi.org/10.32404/rean.v6i3.3324 . Sadak, and Dawood. 2014. “Role of Ascorbic Acid and { $ }\alpha{\ $ }tocopherol in Alleviating Salinity Stress on Flax Plant (Linum Usitatissimum L.).” Journal of Stress Physiology {&} Biochemistry 10 (1): 93–111. Sadak, Mervat Sh, Asmaa R Abd El-Hameid, Faten S A Zaki, Mona G Dawood, and Mohamed E El-Awadi. 2020. “Physiological and Biochemical Responses of Soybean (Glycine Max L.) to Cysteine Application under Sea Salt Stress.” Bulletin of the National Research Centre 44 (1): 1–10. Sadiq, Muhammad, NUDRAT AISHA Akram, and Muhammad Ashraf. 2017. “Foliar Applications of Alpha-Tocopherol Improves the Composition of Fresh Pods of Vigna Radiata Subjected to Water Deficiency.” Turkish Journal of Botany 41 (3): 244–52. Sairam, R K. 1994. “Effects of Homobrassinolide Application on Plant Metabolism and Grain Yield under Irrigated and Moisture-Stress Conditions of Two Wheat Varieties.” Plant Growth Regulation 14 (2): 173–81. Sairam, R K, and Aruna Tyagi. 2004. “Physiology and Molecular Biology of Salinity Stress Tolerance in Plants.” Current Science , 407–21. Sakanaka, Senji, Yumi Tachibana, and Yuki Okada. 2005. “Preparation and Antioxidant Properties of Extracts of Japanese Persimmon Leaf Tea (Kakinoha-Cha).” Food Chemistry 89 (4): 569–75. Sauvesty, A, F Page, and J Huot. 1992. “A Simple Method for Extracting Plant Phenolic Compounds.” Canadian Journal of Forest Research 22 (5): 654–59. Semida, W M, R S Taha, M T Abdelhamid, and M M Rady. 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. Semida, Wael M, Taia A Abd El-Mageed, Saad M Howladar, Mostafa M Rady, and others. 2016. “Foliar-Applied Alpha-Tocopherol Enhances Salt-Tolerance in Onion Plants by Improving Antioxidant Defence System.” Australian Journal of Crop Science 10 (7): 1030–39. Sestak, Zdenek, J Catsky, and Paul Gordon Jarvis. 1971. “Determination of Chlorophyll a and b.In Z Sestak, J Catsky, PG Jarvis, Eds,.” Plant Photosynthetic Production. Manual of Methods.The Hague, Netherlands, Dr. W. Junk NV, Pp , 672–97. Shabala, Lana, Tracey A Cuin, Ian A Newman, and Sergey Shabala. 2005. “Salinity-Induced Ion Flux Patterns from the Excised Roots of Arabidopsis Sos Mutants.” Planta 222 (6): 1041–50. Shaddad, M A, A F Radi, A M Abdel-Rahman, and M M Azooz. 1990. “Response of Seeds of Lupinus Termis and Vicia Faba to the Interactive Effect of Salinity and Ascorbic Acid or Pyridoxine.” Plant and Soil 122 (2): 177–83. Shafiq, Sidra, Nudrat Aisha Akram, Muhammad Ashraf, and Amara Arshad. 2014. “Synergistic Effects of Drought and Ascorbic Acid on Growth, Mineral Nutrients and Oxidative Defense System in Canola (Brassica Napus L.) Plants.” Acta Physiologiae Plantarum 36 (6): 1539–53. Shahbaz, Muhammad, Noor Fatima, Zia Ur Rehman Mashwani, Abida Akram, Ehsan ul Haq, Asma Mehak, Fozia Abasi, et al. 2022. “Effect of Phytosynthesized Selenium and Cerium Oxide Nanoparticles on Wheat (Triticum Aestivum L.) against Stripe Rust Disease.” Molecules 27 (23). https://doi.org/10.3390/molecules27238149 . Shahzadi, Iram, Muhammad Iqbal, Rizwan Rasheed, M Arslan Ashraf, Shagufta Perveen, and Makhdoom Hussain. 2017. “Foliar Application of Selenium Increases Fertility and Grain Yield in Bread Wheat under Contrasting Water Availability Regimes.” Acta Physiologiae Plantarum 39 (8): 1–11. Shalaby, Tarek, Yousry Bayoumi, Tarek Alshaal, Nevien Elhawat, Attila Sztrik, and Hassan El-Ramady. 2017. “Selenium Fortification Induces Growth, Antioxidant Activity, Yield and Nutritional Quality of Lettuce in Salt-Affected Soil Using Foliar and Soil Applications.” Plant and Soil 421 (1–2): 245–58. Sharma, Pallavi, Ambuj Bhushan Jha, Rama Shanker Dubey, and Mohammad Pessarakli. 2012. “Reactive Oxygen Species, Oxidative Damage, and Antioxidative Defense Mechanism in Plants under Stressful Conditions.” Journal of Botany 2012: 1–26. https://doi.org/10.1155/2012/217037 . Shekari, Fariborz, Amin Abbasi, and Seyed Hamid Mustafavi. 2017. “Effect of Silicon and Selenium on Enzymatic Changes and Productivity of Dill in Saline Condition.” Journal of the Saudi Society of Agricultural Sciences 16 (4): 367–74. Smirnoff, Nicholas. 1995. Environment and Plant Metabolism: Flexibility and Acclimation . BIOS Scientific publishers. Solecka, Danuta, and Alina Kacperska. 2003. “Phenylpropanoid Deficiency Affects the Course of Plant Acclimation to Cold.” Physiologia Plantarum 119 (2): 253–62. Spicher, Livia, Gaetan Glauser, and Felix Kessler. 2016. “Lipid Antioxidant and Galactolipid Remodeling under Temperature Stress in Tomato Plants.” Frontiers in Plant Science 7: 167. Stepien, Piotr, and Grazyna Klobus. 2005. “Antioxidant Defense in the Leaves of C3 and C4 Plants under Salinity Stress.” Physiologia Plantarum 125 (1): 31–40. Studier, F William. 1973. “Analysis of Bacteriophage T7 Early RNAs and Proteins on Slab Gels.” Journal of Molecular Biology 79 (2): 237–48. Sullivan, Ch Y. 1979. “Selection for Drought and Heat Tolerance in Grain Sorghum.” Stress Physiology in Crop Plants . Sun, Z W, L K Ren, J W Fan, Q Li, K J Wang, M M Guo, L Wang, et al. 2016. “Salt Response of Photosynthetic Electron Transport System in Wheat Cultivars with Contrasting Tolerance.” Plant, Soil and Environment 62 (11): 515–21. Suo, Jinwei, Qi Zhao, Lisa David, Sixue Chen, and Shaojun Dai. 2017. “Salinity Response in Chloroplasts: Insights from Gene Characterization.” International Journal of Molecular Sciences 18 (5): 1011. Szabados, Laszlo, and Arnould Savoure. 2010. “Proline: A Multifunctional Amino Acid.” Trends in Plant Science 15 (2): 89–97. Tanaka, Yoshiyuki, Makoto Matsuoka, Naoki Yamanoto, Yuko Ohashi, Yuriko Kano-Murakami, and Yoshihiro Ozeki. 1989. “Structure and Characterization of a CDNA Clone for Phenylalanine Ammonia-Lyase from Cut-Injured Roots of Sweet Potato.” Plant Physiology 90 (4): 1403–7. Tavili, A, S Zare, A Enayati, and others. 2009. “Hydropriming, Ascorbic and Salicylic Acid Influence on Germination of Agropyron Elongatum Host. Seeds under Salt Stress.” Research Journal of Seed Science 2 (1): 16–22. Terzi, Rabiye, Neslihan Saruhan Güler, Funda Gül Güven, and Asim Kadioglu. 2018. “Alpha Lipoic Acid Treatment Induces the Antioxidant System and Ameliorates Lipid Peroxidation in Maize Seedlings under Osmotic Stress.” Archives of Biological Sciences 70 (3): 503–11. https://doi.org/10.2298/ABS171218011T . Thipyapong, Piyada, and John C Steffens. 1997. “Tomato Polyphenol Oxidase (Differential Response of the Polyphenol Oxidase F Promoter to Injuries and Wound Signals).” Plant Physiology 115 (2): 409–18. Wahid, Abdul, and Alia Ghazanfar. 2006. “Possible Involvement of Some Secondary Metabolites in Salt Tolerance of Sugarcane.” Journal of Plant Physiology 163 (7): 723–30. Wakeel, Abdul, Muhammad Farooq, Manzoor Qadir, and Sven Schubert. 2011. “Potassium Substitution by Sodium in Plants.” Critical Reviews in Plant Sciences 30 (4): 401–13. Wang, Xiaoyuan, and Peter J Quinn. 2000. “The Location and Function of Vitamin E in Membranes.” Molecular Membrane Biology 17 (3): 143–56. Wu, Nan, Yuangang Zu, Yujie Fu, Yu Kong, Jintong Zhao, Xiaojuan Li, Ji Li, Michael Wink, and Thomas Efferth. 2010. “Antioxidant Activities and Xanthine Oxidase Inhibitory Effects of Extracts and Main Polyphenolic Compounds Obtained from Geranium Sibiricum L.” Journal of Agricultural and Food Chemistry 58 (8): 4737–43. Yu, Xiao-Zhang, and Ji-Dong Gu. 2013. “Phyto-Transport and Assimilation of Selenium.” In Plant-Based Remediation Processes , 159–75. Springer. Zeid, I M, ZFAE Gharib, S M Ghazi, and E Z Ahmed. 2019. “Promotive Effect of Ascorbic Acid, Gallic Acid, Selenium and Nano-Selenium on Seed Germination, Seedling Growth and Some Hydrolytic Enzymes Activity of Cowpea (Vigna Unguiculata) Seedling.” J Plant Physiol Pathol 7 1: 1–8. Zong, Xiao-juan, Da-peng Li, Ling-kun Gu, De-quan Li, Li-xia Liu, and Xiao-li Hu. 2009. “Abscisic Acid and Hydrogen Peroxide Induce a Novel Maize Group C MAP Kinase Gene, ZmMPK7, Which Is Responsible for the Removal of Reactive Oxygen Species.” Planta 229 (3): 485–95. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8692316","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":593745271,"identity":"9429e3e3-a37f-4525-98f4-e0f8c25fd0df","order_by":0,"name":"Radwan Khalil","email":"","orcid":"","institution":"Benha University","correspondingAuthor":false,"prefix":"","firstName":"Radwan","middleName":"","lastName":"Khalil","suffix":""},{"id":593745276,"identity":"bc17046c-67e6-4db9-aebc-1b8f1ab42892","order_by":1,"name":"Amina Gamal","email":"","orcid":"","institution":"Benha University","correspondingAuthor":false,"prefix":"","firstName":"Amina","middleName":"","lastName":"Gamal","suffix":""},{"id":593745280,"identity":"d52cb68b-4ca5-4827-80e8-ca69cbe57c39","order_by":2,"name":"Mohammad Yusuf","email":"","orcid":"","institution":"United Arab Emirates University","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"","lastName":"Yusuf","suffix":""},{"id":593745281,"identity":"1157aa76-0415-4977-acc6-bc50b28cb212","order_by":3,"name":"Samia Haroun","email":"","orcid":"","institution":"Mansoura University","correspondingAuthor":false,"prefix":"","firstName":"Samia","middleName":"","lastName":"Haroun","suffix":""},{"id":593745282,"identity":"523c1fd4-b4bd-4e67-b4dd-b9b04f533e20","order_by":4,"name":"Metwally Rashad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9klEQVRIiWNgGAWjYHACxgMgooG9Aco/QIQeiBaeA0hc4rRIJBCpRbeB+cHBHxU2shtuvj384WcbgxzfjQTmDz/waDE7wGZwmOdMmvGG23lpkr1tDMaSNxLYJHvwamEwOMzYdjhxw+0cMwbeNobEDUAtDDx4tbB/OPjz3//EDTfPGH/828ZQD9TC/PEPXi08Bgd4Gw4ADecxkAbakmBwI4FBGq8th3kKDvMcSzaeeSYvTVrmnIThzDMP26Rl8Gk53r7x4Y8aO9m+42cPf3xTZiPPdzwZyMCjhYEZSiscADtGggEUR/g0IIB8Az73j4JRMApGwYgGACYvWLC2/TxmAAAAAElFTkSuQmCC","orcid":"","institution":"Prince Sattam bin Abdulaziz University","correspondingAuthor":true,"prefix":"","firstName":"Metwally","middleName":"","lastName":"Rashad","suffix":""},{"id":593745283,"identity":"44b032b8-c3b5-4207-b635-1d3af5ae7988","order_by":5,"name":"Hamada A. Nayel","email":"","orcid":"","institution":"Prince Sattam bin Abdulaziz University","correspondingAuthor":false,"prefix":"","firstName":"Hamada","middleName":"A.","lastName":"Nayel","suffix":""},{"id":593745284,"identity":"83b44c16-a10f-4155-8a63-110e6851d069","order_by":6,"name":"Mustafa Abdul Salam","email":"","orcid":"","institution":"Prince Sattam bin Abdulaziz University","correspondingAuthor":false,"prefix":"","firstName":"Mustafa","middleName":"Abdul","lastName":"Salam","suffix":""},{"id":593745285,"identity":"ba9e82c5-d0ff-4865-8f4f-0ab9a677c5b8","order_by":7,"name":"Mohammed Rahmath","email":"","orcid":"","institution":"Prince Sattam bin Abdulaziz University","correspondingAuthor":false,"prefix":"","firstName":"Mohammed","middleName":"","lastName":"Rahmath","suffix":""},{"id":593745286,"identity":"e8cf7329-71ee-4851-aea9-a082d119a40e","order_by":8,"name":"Yousef Ibrahim Daradkeh","email":"","orcid":"","institution":"Prince Sattam bin Abdulaziz University","correspondingAuthor":false,"prefix":"","firstName":"Yousef","middleName":"Ibrahim","lastName":"Daradkeh","suffix":""},{"id":593745287,"identity":"6d0a5aef-af25-4349-9bd9-4d4dbbf6ec51","order_by":9,"name":"Fardous Bassuony","email":"","orcid":"","institution":"Benha University","correspondingAuthor":false,"prefix":"","firstName":"Fardous","middleName":"","lastName":"Bassuony","suffix":""}],"badges":[],"createdAt":"2026-01-25 12:23:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8692316/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8692316/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":103506034,"identity":"efdd4fbe-37f0-44bd-94d9-da434f4a03f6","added_by":"auto","created_at":"2026-02-26 13:33:53","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":238341,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on the morphological traits (a) shoot length, (b) root length, (c) shoot fresh mass, (d) root fresh mass, (e) shoot dry mass, and (f) root dry mass of 40 days old maize (\u003cem\u003eZea mays L.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/fc7800c5008f6d6a53ebccf5.png"},{"id":103257771,"identity":"8707d2e5-acda-4326-862a-b5dfa959598f","added_by":"auto","created_at":"2026-02-23 17:21:52","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":238452,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on (a) leaf area per plant, (b) chlorophyll a, (c) chlorophyll b, and (d) carotenoids of 40 days old maize (\u003cem\u003eZea mays L.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/b48a9cc35f60a0bd8d36ec5f.png"},{"id":103257773,"identity":"b0e6c089-c19a-4519-9cda-a69454751993","added_by":"auto","created_at":"2026-02-23 17:21:52","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":632717,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on (a) electrolyte leakage, (b) membrane stability index, (c) Lipid peroxidation, and (d) proline content of 40 days old maize (\u003cem\u003eZea mays \u003c/em\u003eL\u003cem\u003e.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/cc30a1b323028814419061dd.jpeg"},{"id":103257775,"identity":"c70a8ffc-423f-4c52-885b-7e5a59678df8","added_by":"auto","created_at":"2026-02-23 17:21:52","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":251024,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on (a) total carbohydrate content, (b) total phenolic content, (c) total flavonoid content, and (d) total protein content of 40 days old maize (\u003cem\u003eZea maysL.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/7ad01a8cd380d5d6f9dd9480.png"},{"id":103257770,"identity":"02dac18e-f35b-49d2-87d6-4e61e4eb1633","added_by":"auto","created_at":"2026-02-23 17:21:52","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":337325,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on (a) amylase activity, (b) protease activity, (c) phenylalanine ammonia lyase activity, (d) polyphenol oxidase activity, (e) peroxidase activity, and (f) catalase activity of 40 days old maize (\u003cem\u003eZea mays L.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means ± standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P ≤ 0.05 level according to Tukey’s test.\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/89288588a8c0f890512bb81d.png"},{"id":103505504,"identity":"4d78615b-7c88-40b6-9ed0-4fbc4a7c949a","added_by":"auto","created_at":"2026-02-26 13:31:31","extension":"jpeg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":392395,"visible":true,"origin":"","legend":"\u003cp\u003e(A):\u0026nbsp; Effect of ascorbic acid on protein pattern by SDS-PAGE of maize (\u003cem\u003eZea mays\u003c/em\u003e L.) leaf grown under varied levels of NaCl. Each lane contains equal amounts of protein extracted from maize leaves.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Lane M - protein markers; Lane 1 - control (H\u003csub\u003e2\u003c/sub\u003eO); Lane 2 - 100 mM NaCl; Lane 3 - 150 mM NaCl; Lane 4 - 200 mM NaCl; Lane 5 -\u0026nbsp;\u0026nbsp; 250ppm Ascorbic acid; Lane 6 -\u0026nbsp; 100 mM NaCl + 250ppm Ascorbic acid; Lane 7 -\u0026nbsp;\u0026nbsp; 150 mM NaCl + 250ppm Ascorbic acid; Lane 8 -\u0026nbsp; 200 mM NaCl + 250ppm Ascorbic acid\u003c/p\u003e\n\u003cp\u003e(B): Effect of sodium selenate and α-tocopherol on protein pattern by SDS-PAGE of maize (\u003cem\u003eZea mays L\u003c/em\u003e.) leaf grown under varied levels of NaCl. Each lane contains equal amounts of protein extracted from maize leaves.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Lane M\u0026nbsp;\u0026nbsp; protein markers; Lane 1 - control (H\u003csub\u003e2\u003c/sub\u003eO); Lane 2 - 100 mM NaCl; Lane 3 - 150 mM NaCl; Lane 4 - 200 mM NaCl; Lane 5 - 0.5 μM sodium selenate; Lane 6 - 100 mM NaCl + 0.5μM sodium selenate; Lane 7 - 150 mM NaCl + 0.5μM sodium selenate; Lane 8 - 200 mM NaCl + 0.5μM sodium selenate; Lane 9 - 200ppm α-tocopherol; Lane 10 - 100 mM NaCl+200ppm α-tocopherol; Lane 11 - 150 mM NaCl +200ppm α-tocopherol; Lane 12 - 200mM Nacl +200ppm α-tocopherol\u003c/p\u003e","description":"","filename":"floatimage6.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/30ce9b142d9cc48b6749672f.jpeg"},{"id":103510789,"identity":"b741604b-f69f-41cb-ae46-744f19191652","added_by":"auto","created_at":"2026-02-26 14:07:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3930412,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8692316/v1/256b1ac8-bc4d-4fed-beab-8f1c1628bc1b.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Differential Modulation of Antioxidant Defense and Salinity Tolerance in Maize by Selenium, α-Tocopherol, and Ascorbic Acid","fulltext":[{"header":"Introduction","content":"\u003cp\u003eCrop production has not kept up with the growing need for food, and global agriculture is today confronted with the daunting task of producing almost 70% more food to fulfil the demands of a fast expanding population. Over 6% of the world\u0026rsquo;s arable land is affected by salinity (Parvaiz et al., 2008), with saline soils predominantly distributed across arid and semi-arid regions of Asia (e.g., South and Central Asia, the Middle East), Africa (notably North Africa), Australia, and parts of the Mediterranean basin, as well as irrigated agricultural lands in North and South America. Although several of these regions are not traditionally major maize growing areas, maize was selected for this study because of its global economic importance, its expanding cultivation into marginal and saline-prone environments due to climate change and irrigation-induced salinization, and its well-documented sensitivity to salt stress. Consequently, maize serves as an appropriate model crop for evaluating stress-mitigation strategies aimed at improving crop resilience under saline conditions. This issue is getting worse all across the world, mostly because of the usage of subpar irrigation water and insufficient drainage infrastructure. Salt stress negatively impacts subsequent plant growth and developmental processes in addition to seed germination (Nakashima et al., 2000; Dash and Panda, 2001). Excessive salt accumulation leads to ionic imbalance and osmotic stress, which together disrupt cellular homeostasis and can ultimately result in plant death (Yu and Gu 2013; Gong et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Furthermore, excess of Cl\u003csup\u003e+\u003c/sup\u003e and Na\u003csup\u003e+\u003c/sup\u003e ions buildup in tissues of plants caused by stress caused by salt interferes with vital physiological and metabolic functions and upsets normal ionic homeostasis (Arif et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) The cellular Na\u003csup\u003e+\u003c/sup\u003e/K\u003csup\u003e+\u003c/sup\u003e ratio, which is essential for preserving membrane potential and enzymatic activity, is also disrupted by this imbalance. Plants have, of course, created a variety of adaptive techniques to lessen the adverse consequences of various abiotic stressors.\u003c/p\u003e \u003cp\u003eAlthough it can be hazardous to the environment, selenium serves as a necessary trace element for both people and animals. Its beneficial and harmful effects are closely separated and rely on a number of variables, including its chemical form, concentration, and environmental circumstances (Yu and Gu, 2013). Humans, plants, animals, and microbes all contain trace levels of selenium. Despite being needed in very small amounts, it is an essential element that has been demonstrated to increase a plant's resilience to oxidative damage brought on by UV light, postpone senescence, and encourage the growth and vitality of ageing seedlings (Rostami and Abbaspour 2019).\u003c/p\u003e \u003cp\u003eThe low-molecular-weight lipophilic antioxidant α-tocopherol, also referred to as vitamin E, is produced spontaneously by organisms that use green photosynthesis (Munn\u0026eacute;-Bosch and Alegre 2000; Munn\u0026eacute;-Bosch \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Munn\u0026eacute;-Bosch \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Falk and Munn\u0026eacute;-Bosch 2010). Plant membranes are the primary location for both α- and γ-tocopherol. In particular, α-tocopherol is the primary type found in plant leaves' membranes involved in photosynthetic processes, while γ-tocopherol is mostly found in tissues that are not photosynthetic, such as nuts, seeds, and fruits (Grilo et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Although it may also occur, to a lesser extent, in other cellular compartments including the cytoplasm and vacuoles, tocopherol production mostly takes place within chloroplasts\u0026mdash;more precisely, in the plastoglobuli, thylakoid membranes, and envelope (Rautenkranz et al. \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Li et al. 2008) α-Tocopherol is essential for shielding plants from cellular oxidation brought on by stress. Exogenous α-tocopherol injection has been demonstrated to successfully enhance plant growth and developmental processes across a range of adverse environmental conditions (Sadiq, Akram, and Ashraf 2017; Khalil et al. \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Because it is prevalent in cellular membranes, α-tocopherol prevents lipid peroxidation and scavenges reactive free radicals, preserving membrane integrity under stress (Munn\u0026eacute;-Bosch \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Tocopherols not only shield cell membranes but also control the production of particular genes that respond to stress, which helps to modify plant defence and adaption systems (Munn\u0026eacute;-Bosch \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Li et al. 2008; Suo et al. \u003cspan citationid=\"CR119\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAscorbic acid, commonly referred to as vitamin C, is a water-soluble antioxidant, is an essential substrate for the enzymatic detoxification of hydrogen peroxide. By controlling cell development and division, it encourages plant growth and improves resistance to abiotic challenges including salinity and temperature extremes (Smirnoff \u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e1995\u003c/span\u003e). By increasing plant resistance to salinity, drought, and pests, seed priming improves crop performance in stressful environmental conditions (Harris et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). This method eventually increases output by assisting crop plants in overcoming stressors like drought and pest damage ( Harris et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). According to (David Harris et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) The nutrient content of seeds is improved by nutrient seed priming, germination as well as seedling establishment stressful by soaking seeds in nutrient solutions rather than water. This study aims to elucidate the potential role of seed priming with ascorbic acid, α-tocopherol, and selenium in enhancing salinity tolerance of maize. The research explores how these treatments modulate plant growth, physiological attributes, antioxidant defense mechanisms, and nutrient homeostasis under varying salinity levels, ultimately contributing to improved crop performance and yield stability under salt stress conditions.\u003c/p\u003e"},{"header":"Materials and techniques","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant material\u003c/h2\u003e \u003cp\u003eWe purchased pure Seeds of maize (\u003cem\u003eZea mays\u003c/em\u003e L.) (TWC 321) from the Ministry of Agriculture's Agricultural Research Centre in Giza, Egypt.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eChemical source and its preparation\u003c/h3\u003e\n\u003cp\u003eSigma\u0026ndash;Aldrich (USA) provided the sodium chloride (NaCl), ascorbic acid, sodium selenium, and α-tocopherol. Based on initial tests, working solutions of ascorbic acid (250 ppm), α-tocopherol (200 ppm), and sodium selenate (0.5 \u0026micro;M) were made. In order to create salinity stress, sodium chloride was employed as the salt source at concentrations of 100, 150, and 200 mM.\u003c/p\u003e\n\u003ch3\u003eDesign of experiments and treatment regimens\u003c/h3\u003e\n\u003cp\u003eThe study was conducted in a greenhouse at Benha University's Faculty of Science's Botany and Microbiology Department in Egypt, using 80 plastic pots (25 \u0026times; 40 cm) filled with a clay\u0026ndash;sand combination (2:1, v/v). Five replicates of a completely randomised block design were used for the treatments. Five seeds per pot were planted after surface-sterilized Zea mays L. seeds were soaked for 12 hours in either sodium selenate (0.5 \u0026micro;M), α-tocopherol (200 ppm), ascorbic acid (250 ppm), or deionised water (control). With soil moisture kept at 80% field capacity, Plants were exposed to 0, 100, 150, and 200 mM NaCl at 21 days. Samples for growth, physiological, biochemical, and SDS-PAGE protein analysis were gathered at 40 days.\u003c/p\u003e\n\u003ch3\u003eGrowth characteristics\u003c/h3\u003e\n\u003cp\u003eSamples of plants were split into roots, stems, and leaves after being carefully cleaned with distilled water to get rid of any remaining soil particles. Recorded were the main root and stem lengths, each plant's leaf count and leaf area (cm\u0026sup2;). Following the measurement of the fresh weights of the roots and stems, every sample was dried in an oven at 70\u0026deg;C in order to calculate their dry weight (g/plant). For ensuing biochemical investigations, tissues that had been dried were pulverised into a fine powder and stored dry.\u003c/p\u003e\n\u003ch3\u003eIdentifying and extracting photosynthetic pigments\u003c/h3\u003e\n\u003cp\u003eThe process described by was used to extract and find out the total amount of photosynthetic pigments Fadeel (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1962\u003c/span\u003e). Anhydrous 100% acetone was used to remove a known fresh weight of the topmost completely inflated leaves, which were then stored at -5\u0026deg;C for at least 12 hours. Using cold acetone and acid-washed sand in a mortar and pestle, the tissues were ground into a homogeneous mixture on ice. Anhydrous sodium sulphate was added to help with dehydration during grinding, and a tiny bit of magnesium carbonate was added to balance the acidity of the tissue. After 15 minutes of centrifuging the homogenate at 4000 \u0026times; g, the residue was extracted again using cold acetone to remove all of the pigment. 100% acetone was used to make up the combined extracts to a given volume. A Perkin-Elmer double-beam spectrophotometer (Model 200\u0026ndash;20) was used to detect absorbance at 440.5, 644, and 662 nm. The amounts of carotenoid, chlorophyll a, and chlorophyll b were determined using conventional formulas for extracts made entirely of acetone (Sestak \u003cem\u003eet al\u003c/em\u003e., 1971):\u003c/p\u003e \u003cp\u003eChlorophyll a\u0026thinsp;=\u0026thinsp;9.78 E662- 0.99 E644 (\u0026micro;g /ml)\u003c/p\u003e \u003cp\u003eChlorophyll b\u0026thinsp;=\u0026thinsp;21.4 E644\u0026ndash;4.65 E662 (\u0026micro;g /ml)\u003c/p\u003e \u003cp\u003eChlorophyll (a\u0026thinsp;+\u0026thinsp;b)\u0026thinsp;=\u0026thinsp;5.13 E662\u0026thinsp;+\u0026thinsp;20.41 E644 (\u0026micro;g /ml)\u003c/p\u003e \u003cp\u003eCarotenoids\u0026thinsp;=\u0026thinsp;4.69 E440.5 \u0026minus;\u0026thinsp;0.268 chlorophyll (a\u0026thinsp;+\u0026thinsp;b) (\u0026micro;g /ml)\u003c/p\u003e \u003cp\u003eThe outcomes of the aforementioned equations were converted to mg pigment g⁻\u0026sup1; fresh weight after being expressed as \u0026micro;g pigment ml⁻\u0026sup1; extracts (Fadeel, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1962\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eStress biomarkers (electrolyte leakage (EL), membrane stability index (MSI) lipid-peroxidation (Malondialdehyde; MDA)\u003c/h2\u003e \u003cp\u003eThe technique outlined by was used to measure the total amount of inorganic ions that seeped out of the leaves Sullivan (\u003cspan citationid=\"CR117\" class=\"CitationRef\"\u003e1979\u003c/span\u003e). Leaf discs were boiled at 100\u0026deg;C (EC\u0026#120460;) after being incubated in 10 ml of deionised water at 45\u0026deg;C (ECₐ) and 55\u0026deg;C (EC\u003csub\u003eβ\u003c/sub\u003e) for 30 minutes in order to quantify electrolyte leakage. The calculation for leakage (%) was:\u003c/p\u003e \u003cp\u003eElectrolyte leakage (%) = (ECb-ECa)/ECcX100 where, EC\u0026thinsp;=\u0026thinsp;Electrical Conductivity.\u003c/p\u003e \u003cp\u003eThe MSI was evaluated by incubating two sets of 200 mg of leaf tissue in 10 ml of deionised water. For 30 minutes (C₁), the initial set was heated to 40\u0026deg;C, and for 10 minutes (C₂), boiling the second set was done at 100\u0026deg;C. A conductivity meter was used to record electrical conductivities. MSI was computed using the formula outlined by (Sairam, \u003cspan citationid=\"CR97\" class=\"CitationRef\"\u003e1994\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMSI (%) = [1 – (C1/C2) × 100]\u003c/h3\u003e\n\u003cp\u003eThe degree of lipid peroxidation was measured using the method of malondialdehyde (MDA) content Hodges et al. (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). After homogenising two grammes of leaf tissue in ten millilitres of 5% thiobarbituric acid (TBA), centrifuging the mixture for ten minutes at 15,000 \u0026times; g. 4ml of 0.5% TBA in 20% trichloroacetic acid (TCA) were added to two millilitres of the supernatant. The mixture was rapidly chilled in an ice bath after being heated to 95\u0026deg;C for 30 minutes and centrifuged once more for 10 minutes at 10,000 \u0026times; g. Non-specific absorbance at 600 nm was deducted from the supernatant's absorbance, which was measured at 532 nm. To calculate the MDA concentration, an extinction coefficient of 155 mM⁻\u0026sup1; cm⁻\u0026sup1; was employed, which was then reported as \u0026micro;g MDA g⁻\u0026sup1; fresh weight.\u003c/p\u003e\n\u003ch3\u003eMeasurement and extraction of proline\u003c/h3\u003e\n\u003cp\u003eThe calculation of free proline was done using (Bates et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e1973\u003c/span\u003e). It was fresh tissue homogenised in Sulfosalicylic acid (3%), the filtrate was reacted with Ninhydrin acid, and toluene was used to extract the amount of proline. Proline concentration was established using a standard curve and represented as mg proline 100 g⁻\u0026sup1; dry weights after absorbance was measured at 520 nm.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003ePhenolic compounds' extraction and identification\u003c/h2\u003e \u003cp\u003eThe process of extracting phenolic compounds followed the protocol described by Sauvesty et al. (\u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). 70% ethanol was used to extract phenolic chemicals from dry powdered tissue over the course of an overnight period at 40\u0026deg;C. After centrifuging and low-pressure evaporation, the extract was diluted with distilled water. Applying the Folin-Ciocalteu technique, one millilitre of The sample was hydrolysed with 2 N HCl to liberate bound phenolics, neutralised, and utilised to determine the total amount of phenol (Lowe, \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e1993\u003c/span\u003e) was employed to determine the phenolic aglycone.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eFlavonoid extraction and identification\u003c/h2\u003e \u003cp\u003eAluminium chloride colorimetric analysis was employed to calculate the total flavonoid concentration (Sakanaka \u003cem\u003eet al.\u003c/em\u003e, \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Methanol was utilised to extract the dried materials, and 0.25 millilitres of the clear extract were added to 1.25 millilitres of distilled water. Following the addition of 75 \u0026micro;l of a solution containing 5% sodium nitrite, the mixture was incubated for six minutes. After adding 150 \u0026micro;l of a 10% aluminium chloride solution and letting it stand for five minutes, 0.5 ml of 1 M sodium hydroxide was added. The absorbance was measured at 510 nm after the total volume was adjusted to 2.5 ml using distilled water. To calculate the total flavonoid content, a myricetin standard curve was utilised, which was then reported as mg g⁻\u0026sup1; dry weight.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eProtein extraction and identification\u003c/h2\u003e \u003cp\u003eThe method described by was used to extract proteins Eltayeb et al. (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). To extract the soluble protein, 100 mg of dry powdered tissue was incubated for two hours at 90\u0026deg;C in 10 ml of H\u003csub\u003e2\u003c/sub\u003eO. The mixture was centrifuged for 10 minutes at 5000 g after cooling. After that, the leftover residue was homogenised with 10 millilitres of 1 N NaOH for two hours at 90\u0026deg;C in order to remove the water-insoluble protein. Following a 10-minute centrifugation at 5000 g, HCl was used to neutralise the clear supernatant, and distilled water was added to each extract to reach a given volume.\u003c/p\u003e \u003cp\u003eProtein analysis was done using the modified Folin-Lowry technique that was used by Hartree ( 1972). After combining one millilitre of the transparent extract of protein with 0.9 millilitres of alkaline solution of sodium carbonate, the mixture was heated to 50\u0026deg;C for ten minutes. Following cooling, a solution of potassium sodium tartrate and copper sulphate was added in 0.1 millilitres, and the mixture was allowed to stand at room temperature for ten minutes. The 10% Folin-Ciocalteu reagent (3 ml) was then added and mixed right away. Protein concentration was determined using a bovine serum albumin standard curve and represented as mg g⁻\u0026sup1; dry weight after 30 minutes, when absorbance was measured at 750 nm against a reagent blank.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eProtease and Amylase assays\u003c/h2\u003e \u003cp\u003eThe fresh tissue was homogenised in a pH 7.6 phosphate buffer containing 20 mM to extract protease, and the buffer used was 100 mM acetate (pH 6.0) to extract amylase. It made use of bovine serum albumin (BSA) as the substrate to measure protease activity. After being incubated at 40\u0026deg;C for 60 minutes, the reaction mixture which contained crude extract and BSA solution\u0026mdash;was terminated with 10% trichloroacetic acid. Following centrifugation, the modified Folin\u0026ndash;Lowry method was used to quantify the amount of soluble peptides Hartree (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e1972\u003c/span\u003e). In order to measure amylase activity, half a millilitre of crude extract and half a millilitre of 0.5% soluble starch were incubated for 30 minutes at 40\u0026deg;C in 0.1 M acetate buffer (pH 6.0) with 5 mM CaCl₂. HgCl₂ was used to stop the reaction, and Nelson's technique was used to measure the reducing sugars (Clark and Switzer, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1977\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eAntioxidant enzyme extraction and analysis\u003c/h2\u003e \u003cp\u003eA 100 mM phosphate buffer (pH 7) containing 0.1 g PVP and 0.1 mM Na₂EDTA was used to homogenise 0.25 g of fresh leaf material in liquid nitrogen. After centrifuging the homogenate for 10 minutes at 15,000 \u0026times; g, the supernatant was centrifuged again for 10 minutes at 18,000 \u0026times; g. The final extract was gathered and maintained at 4\u0026deg;C to gauge catalase (CAT) and peroxidase (POD) activity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003ePeroxidase (POX) activity\u003c/h2\u003e \u003cp\u003eGuaiacol was used as the substrate to assess the activity of peroxidase (POX, EC 1.11.1.7). 10 mM phosphate buffer (pH 7.0), 10 mM H\u003csub\u003e2\u003c/sub\u003eO₂, 20 mM guaiacol, and 0.5 ml of the crude enzyme extract were all included in the reaction mixture (Malick and Singh, \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). The increase in absorbance at 470 nm was used to track the production of oxidised guaiacol (Klapheck et al., 1990) use an ultraviolet spectrophotometer (Spectronic 601). The change in optical density g-1 fresh weight min-1 is used to express enzyme activity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eCatalase (CAT) activity\u003c/h2\u003e \u003cp\u003e0.5 millilitres of enzyme extract in the reaction mixture, 30% (w/v) H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, and phosphate buffer (50 mM, pH 7.0) was used to measure CAT activity (EC 1.11.1.6.) (Aebi, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1984\u003c/span\u003e). Catalase activity was measured using a Spectronic 601 UV spectrophotometer as the decrease in absorbance at 240 nm caused by H\u003csub\u003e2\u003c/sub\u003eO₂ breakdown and expressed as \u0026micro;M H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e oxidised g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e fresh weight min-\u003csup\u003e1\u003c/sup\u003e (Havir and McHale 1987).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003ePhenylalanine ammonia-lyase (PAL) determination\u003c/h2\u003e \u003cp\u003ePAL (EC 4.3.1.5) extraction and assay were performed as instructed Solecka and Kacperska (2003). Following a 20-minute centrifugation at 12,000 \u0026times; g at 4\u0026deg;C, fresh leaf tissue was homogenised in liquid nitrogen using extraction buffer (5 mM EDTA; 5 mM ascorbic acid; 50 mM Tris\u0026ndash;HCl, pH 8.9). The PAL assay used the supernatant as a crude enzyme. 100 \u0026micro;l of enzyme extract and 0.9 ml of substrate solution (16 mM L-phenylalanine in 50 mM Tris\u0026ndash;HCl, pH 8.9, with 3.6 mM NaCl) were included in the reaction mixture. It was incubated for one hour at 37\u0026deg;C and stopped with 500 \u0026micro;l of 6 N HCl. At 290 nm, absorbance was measured. nmol trans-cinnamic acid g⁻\u0026sup1; FW h⁻\u0026sup1; was used to express PAL activity, which is the quantity that produces 1 \u0026micro;M trans-cinnamic acid every hour.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of polyphenol oxidase (PPO)\u003c/h2\u003e \u003cp\u003ePolyphenol oxidase (EC 1.14.18.1) was extracted as described by Kar and Mishra (1976) with slight modification. After being homogenised in ice-cold 0.1 M phosphate buffer (pH 7.0) at 4\u0026deg;C, Samples of frozen tissue were centrifuged at 10,000 \u0026times; g for 10 minutes. Polyphenol oxidase (PPO) activity was measured using the resultant supernatant using the procedure of Nguyen et al. (\u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). 2.5 ml of substrate solution (0.05 M phosphate buffer) and 0.5 ml of crude enzyme extract, pH 6.0, with Catechol at 0.05 M) made up the assay combination, which was then incubated for 30 minutes at 30\u0026deg;C. PPO activity was reported as ABS unit fresh weight h⁻\u0026sup1; g⁻\u0026sup1;, and absorbance was measured at 420 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eProtein pattern analysis by gel electrophoresis\u003c/h2\u003e \u003cp\u003eSDS-PAGE was employed to separate protein samples on a 12% polyacrylamide gel. According to their size, negatively charged proteins in this technology move through the gel matrix in the direction of the anode under an electric field by using the Laemmli (\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e1970\u003c/span\u003e) as edited by Studier (\u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e1973\u003c/span\u003e) and equal amounts of isolated proteins were loaded.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eGel running and staining\u003c/h2\u003e \u003cp\u003eAfter combining protein samples with an equivalent amount of sample buffer, they were denatured for three to five minutes at 80 to 90 degrees Celsius, cooled on ice, and then placed onto the gel. Using 1\u0026times; Tris/glycine\u0026ndash;SDS running buffer, electrophoresis was carried out at 80 V for the stacking gel and 100 V for the separating gel.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eDetection of protein bands\u003c/h2\u003e \u003cp\u003eSDS-PAGE 200 ml of the gels was stained overnight with Coomassie Brilliant Blue R-250, and the distaining solution (200 ml) was utilised to distain them for two hours while gently shaking them. This process was repeated until the background was clear. Using Gel Pro Analyser version 3 software, separated protein bands were scanned and examined using a Gel Documentation System (GDS) to compare molecular weights, band intensity, and polypeptide patterns to common protein markers.\u003c/p\u003e \u003cp\u003eExtraction and estimation of certain minerals:\u003c/p\u003e \u003cp\u003eIn an oven set to 80˚C, plant materials were dried until their weight remained constant. The dry material was broken down using the technique of Chapman and Pratt (1962). A 250 ml flask for digestion that had been cleaned with acid was filled with a 0.2 g sample of ground plant material. On a heater, the sample was broken down till dense white fumes formed and the solution cleared to around 2.5 millilitres when six millilitres of a concentrated sulfuric\u0026ndash;perchloric acid mixture (70%, 5:1 v/v) was added. A 50 ml volumetric flask was used to quantitatively transfer the digest once it had cooled. It was then diluted to volume using Whatman No. 42 paper was used to filter the distilled water. The filtrate was kept for K, Ca, Na, Mg, and Se analyses. Using a flame photometer, potassium was measured by Ranganna (1977) and atomic absorption of Ca, Na, mg, and Se using the method described in Julshamn \u003cem\u003eet al\u003c/em\u003e. (\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2005\u003c/span\u003e)\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eIBM SPSS Statistics for Windows, Version 19.0, was used to statistically analyse the experiment's data using analysis of variance (ANOVA). IBM Corp., Armonk, NY, and displayed as treatment mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SE. The least significant difference (LSD) at p\u0026thinsp;\u0026le;\u0026thinsp;0.05 was used to compare the treatment means. The LSD test indicates that bars with the same letter are not substantially different at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec25\" class=\"Section2\"\u003e \u003ch2\u003eGrowth performance\u003c/h2\u003e \u003cp\u003e When maize plants were exposed to salt stress and those that were not, their responses to three distinct growth regulators (selenium, α-tocopherol, and ascorbic acid) varied in terms of their growth performance (length of shoot and root; fresh mass of shoot and root; dry mass of shoot and root; leaf area; Figs.\u0026nbsp;1A-F and Fig.\u0026nbsp;2A). When compared to the untreated (water-soaked) control plants, the pre-sowing seed soaking treatment of Selenium, α-tocopherol, and ascorbic acid significantly increased the following attributes: shoot length (4.4, 8.5, and 8.97%), root length 13.9, 14.3 and 16.19%), shoot fresh mass (1.7, 11.7 and 12.52%), root fresh mass (66.6, 67.1 and 69.33%), shoot dry mass (21.1, 42.1 and 89.47%), root dry mass (24, 52, and 56%), and leaf area (9.2, 25.5 and 27.7%). However, in a concentration-dependent manner, growth performance was dramatically reduced by increasing salt concentrations (0, 50, 100, and 200 mM). In contrast to plants treated with selenium and α-tocopherol, ascorbic acid significantly restored the loss, although seed soaking treatments of selenium and α-tocopherol, and ascorbic acid under salt stress were able to overcome the damage. Plants treated with ascorbic acid through pre-sowing seed soaking treatment demonstrated the highest growth performance of maize plants under salt stress.\u003c/p\u003e \u003cp\u003eFigure\u0026nbsp;1. Effect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on the morphological traits (a) shoot length, (b) root length, (c) shoot fresh mass, (d) root fresh mass, (e) shoot dry mass, and (f) root dry mass of 40 days old maize (\u003cem\u003eZea mays L.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the five replicates (n\u0026thinsp;=\u0026thinsp;5). Means that do not share a letter are significantly different at P\u0026thinsp;\u0026le;\u0026thinsp;0.05 level according to Tukey\u0026rsquo;s test.\u003c/p\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003ePigments and Photosynthetic performance\u003c/h2\u003e \u003cp\u003e When compared to control plants, maize plants examined 40 days after sowing showed a substantial drop in chlorophyll a, b, and carotenoid levels at increasing salt concentrations (0, 50, 100, and 200 mM) (Figs.\u0026nbsp;2B-D). When compared to non-salinized control plants, the largest inhibitory effect of salinity on carotenoid (67%), chlorophyll a (77.1%), and chlorophyll b (64%), was observed at 200 mM NaCl. However, as compared to untreated control plants, treatments with selenium, α-tocopherol, and ascorbic acid considerably enhanced the quantities of photosynthetic pigments. They also recovered the loss caused by the same concentrations of salt in maize plants. Furthermore, ascorbic acid was found to be more promising and to have the greatest protective impact against all concentrations of salt stress in maize plants when compared to selenium, α-tocopherol, and ascorbic acid.\u003c/p\u003e \u003cp\u003eFigure\u0026nbsp;2. Effect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on (a) leaf area per plant, (b) chlorophyll a, (c) chlorophyll b, and (d) carotenoids of 40 days old maize (\u003cem\u003eZea mays L.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the five replicates (n\u0026thinsp;=\u0026thinsp;5). Means that do not share a letter are significantly different at P\u0026thinsp;\u0026le;\u0026thinsp;0.05 level according to Tukey\u0026rsquo;s test.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003eStress Biomarkers [Electrolyte leakage (EL), Membrane stability index (MSI), Lipid peroxidation and proline]\u003c/h2\u003e \u003cp\u003eLipid peroxidation and EL significantly increased when concentrations of NaCl (0, 50, 100, and 200 mM) were applied compared to the control plants. When compared to control plants, the plants exposed to 200 mM NaCl showed the highest levels of electrolyte leakage and lipid peroxidation, which were 50.4% and 104.2%, respectively (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003eA-\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). However, when selenium, α-tocopherol, and ascorbic acid were present, maize plants under NaCl stress shown a considerable decrease in electrolyte leakage of 42%, 48.6%, and 53.3%, respectively. Furthermore, when compared to maize plants exposed to 200 mM salt stress, lipid peroxidation also demonstrated a significant decrease of 54.7%, 69.1%, and 79.4% in the presence of selenium, α-tocopherol, and ascorbic acid, respectively. As maize plants were subjected to 200 mM NaCl, their MSI dropped by 29.4% as compared to control plants (Figs, 3B). However, when tested on plants under NaCl stress, ascorbic acid recovered the loss of MSI by 40.2% compared to plants subjected to 200 mM of NaCl, out of selenium, α-tocopherol, and ascorbic acid.\u003c/p\u003e \u003cp\u003eSalinity stress significantly increased proline accumulation in maize shoots, with a progressive rise at 100\u0026ndash;200 mM NaCl and a maximum increase of 24.2% at 200 mM compared with non-salinized plants (Figs, 3D). Exogenous application of sodium selenate, α-tocopherol, or ascorbic acid further enhanced proline content under salinity. At 100 mM NaCl, proline increased by 23.1%, 18.18%, and 31.81%, respectively, relative to the salinity-stressed control, indicating an enhanced osmotic adjustment under salt stress\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003eNutritional status (Carbohydrates, Phenolics, Flavonoids, and Protein)\u003c/h2\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFigures 4A\u0026ndash;D make it clear that, when compared to control plants, maize plants displayed a significant pattern of change for protein, phenolics, carbohydrates, and flavonoids regardless of treatment patterns. The nutritional status of maize plants was significantly reduced by all applied concentrations of NaCl (0, 50, 100, and 200 mM); however, plants exposed to 200 mM salt stress showed the greatest decreases in carbohydrates (20.03%), phenolics (27.6%), flavonoids (37.5%), and protein (35.3%) when compared to non-salinized control plants. Furthermore, as compared to their respective control or salinized plants, the application of selenium, α-tocopherol, and ascorbic acid through seed soaking enhanced the amounts of total carbohydrates, phenolics, flavonoids, and protein in the maize plant's shoot.\u003c/p\u003e \u003cp\u003eFigure\u0026nbsp;4. Effect of sodium selenate (Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e4\u003c/sub\u003e), a-tocopherol, and ascorbic acid on (a) total carbohydrate content, (b) total phenolic content, (c) total flavonoid content, and (d) total protein content of 40 days old maize (\u003cem\u003eZea maysL.)\u003c/em\u003e plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the five replicates (n\u0026thinsp;=\u0026thinsp;5). Means that do not share a letter are significantly different at P\u0026thinsp;\u0026le;\u0026thinsp;0.05 level according to Tukey\u0026rsquo;s test.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section2\"\u003e \u003ch2\u003eAntioxidant enzymes Peroxidase and Catalase, (Amylases, Proteases, Phenylalanine ammonia- lyase, Polyphenol oxidase activities)\u003c/h2\u003e \u003cp\u003eWhen sodium selenium, α-tocopherol, and ascorbic acid were applied through seeds, the activity of amylases, proteases, and peroxidases was greatly reduced; however, the activities of phenylalanine ammonia-lyase, polyphenol oxidase, and catalase increased in comparison to the plants that were not treated and the plants that were treated with NaCl (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e5\u003c/span\u003eA-F). However, in maize plants, the activities of amylases, proteases, polyphenol oxidase, and peroxidase were all stimulated by salt stress. When compared to the untreated plants, the 200 mM salt treatment had the greatest stimulatory effect on amylases, protease, polyphenol oxidase, and peroxidase activities, increasing them by 60.4%, 64.3%, 58.97%, and 38.14%, respectively. Phenylalanine ammonia-lyase and catalase also decreased as the concentration of salt increased.\u003c/p\u003e \u003c/div\u003e\n\u003cp\u003e\u003cem\u003eProtein Profile\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eSDS-PAGE analysis of maize leaves revealed protein bands ranging from 95.54 to 39.28 kDa across all treatments. In both untreated and salt-stressed plants, three protein bands (95.54, 72.02, and 44.63 kDa) were consistently expressed, regardless of sodium selenate or \u0026alpha;-tocopherol application, suggesting these may represent constitutive or housekeeping proteins. Control plants exhibited six bands (95.54, 72.02, 67.93, 59.29, 44.63, and 39.28 kDa), whereas salt stress alone altered the pattern to four bands (95.54, 72.02, 59.29, and 44.63 kDa), indicating the possible downregulation or degradation of specific stress-sensitive proteins. Treatment with sodium selenate or \u0026alpha;-tocopherol induced the appearance of a new 82.84 kDa band in control plants, suggesting activation of stress-protective proteins, whereas in salt-stressed plants, two bands (59.29 and 39.28 kDa) appeared transiently and then disappeared, which may reflect dynamic protein regulation under combined stress and treatment conditions.\u003c/p\u003e\n\u003cp\u003eSeed priming with 250 ppm ascorbic acid increased the number of protein bands in plants exposed to 100\u0026ndash;200 mM NaCl, indicating enhanced synthesis of stress-responsive proteins. Notably, ascorbic acid treatment led to the de novo synthesis of two polypeptides (80.47 and 67.43 kDa) compared to controls, which displayed three bands (80.47, 67.43, and 34.93 kDa) that disappeared under salt stress. These changes suggest that ascorbic acid may stimulate protective protein expression, potentially including antioxidant enzymes or osmoprotective proteins, enhancing the plant\u0026rsquo;s tolerance to salinity.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eMineral contents\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eIn comparison to control plants, the accumulation of Na+ was 33.3% at the maximum salinity (200 mM NaCl). However, as NaCl concentrations increased, the amounts of K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, and Mg\u003csup\u003e+2\u003c/sup\u003e in maize shoots dropped dramatically, reaching their lowest values in plants treated with 200 mM. These contents were estimated to be 9.25%, 9.79%, 27.20%, 27.63%, and 10.77% lower than those of their respective control plants (Table 1). Ascorbic acid, sodium selenate, and \u0026alpha;-tocopherol significantly altered the levels of Na\u003csup\u003e+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, and Mg+2 when compared to control plants and plants treated to varying concentrations of NaCl. \u0026nbsp;Conversely, ascorbic-soaked seed exhibited the greatest impact on the levels of inorganic cations (Na\u003csup\u003e+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, and Mg\u003csup\u003e+2\u003c/sup\u003e). In comparison to untreated control plants and 200 mM salinized plants, Na\u003csup\u003e+\u003c/sup\u003e decreased by 38.9% and 19.81%, respectively, while K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, and Mg\u003csup\u003e+2\u003c/sup\u003e increased by 31.4%, 43.5%, 108.5%, 127.9%, and 96.2%, respectively, when compared to reference control, and increased by 15.3%, 26.8%, 43.8%, 58.3%, and 57.1% %, respectively, in comparison to plants treated with 200 mM NaCl.\u003c/p\u003e\n\u003cp\u003eTable 1. Effect of sodium selenate (Na2SeO4), a-tocopherol, and ascorbic acid on uptake of (a) Na\u003csup\u003e+\u003c/sup\u003e, (b) K\u003csup\u003e+\u003c/sup\u003e, (c) Ca\u003csup\u003e+\u003c/sup\u003e, (d) K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e, (e) Ca\u003csup\u003e2+\u003c/sup\u003e/Na\u003csup\u003e2+\u003c/sup\u003e, and (f) Mg\u003csup\u003e2+\u003c/sup\u003e of 40 days old maize (Zea mays L.) plants grown under varied levels (100, 150, and 200 mM) of NaCl. Data are means \u0026plusmn; standard error of the five replicates (n = 5). Means that do not share a letter are significantly different at P \u0026le; 0.05 level according to Tukey\u0026rsquo;s test\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"749\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eStage\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatments\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNacl\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003emM\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNa\u003csup\u003e+\u003c/sup\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eK\u003csup\u003e+\u003c/sup\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCa\u003csup\u003e+2\u003c/sup\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eK\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCa\u003csup\u003e+2\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMg\u003csup\u003e+2\u003c/sup\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"16\" style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e40-day- old maize plant\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eReference\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003econtrol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.720\u0026plusmn;0.001\u003csup\u003ei\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.882\u0026plusmn;0.001\u003csup\u003eg\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.245\u0026plusmn;0.002\u003csup\u003eg\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.692\u0026plusmn;0.001\u003csup\u003eg\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.825\u0026plusmn;0.003\u003csup\u003eg\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.958\u0026plusmn;0.002\u003csup\u003ee\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.805\u0026plusmn;0.003\u003csup\u003ej\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.798\u0026plusmn;0.003\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.237\u0026plusmn;0.002\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.640\u0026plusmn;0.003\u003csup\u003ef\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.797\u0026plusmn;0.001\u003csup\u003ef\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.871\u0026plusmn;0.006\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e150\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.371\u0026plusmn;0.005\u003csup\u003eo\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.694\u0026plusmn;0.001\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.111\u0026plusmn;0.001\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.502\u0026plusmn;0.001\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.626\u0026plusmn;0.004\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.801\u0026plusmn;0.004\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e200\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.387\u0026plusmn;0.002\u003csup\u003ep\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.708\u0026plusmn;0.002\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.025\u0026plusmn;0.001\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.504\u0026plusmn;0.000\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.597\u0026plusmn;0.002\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.747\u0026plusmn;0.001\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003esodium selenate 0.5 \u0026micro;M\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.293\u0026plusmn;0.002\u003csup\u003ee\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.891\u0026plusmn;0.003\u003csup\u003eh\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.746\u0026plusmn;0.004\u003csup\u003eh\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.824\u0026plusmn;0.002k\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.197\u0026plusmn;0.002\u003csup\u003el\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.480\u0026plusmn;0.002\u003csup\u003ej\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.371\u0026plusmn;0.001\u003csup\u003ef\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.810\u0026plusmn;0.002\u003csup\u003ee\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.647\u0026plusmn;0.004\u003csup\u003ee\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.763\u0026plusmn;0.003\u003csup\u003ei\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.116\u0026plusmn;0.001\u003csup\u003ej\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.410\u0026plusmn;0.001\u003csup\u003eh\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e150\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.187\u0026plusmn;0.002\u003csup\u003el\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.76\u0026plusmn;0.001\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.511\u0026plusmn;.0.002\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.551\u0026plusmn;0.001\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.787\u0026plusmn;0.003\u003csup\u003ee\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.088\u0026plusmn;0.003\u003csup\u003ef\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e200\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.347\u0026plusmn;0.002\u003csup\u003en\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.802\u0026plusmn;0.003\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.187\u0026plusmn;0.001\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.538\u0026plusmn;0.004\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.653\u0026plusmn;0.006\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.781\u0026plusmn;0.001\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 83px;\"\u003e\n 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\u003cp\u003e\u003cstrong\u003e2.433\u0026plusmn;0.004i\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e200\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.328\u0026plusmn;0.006\u003csup\u003em\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.932\u0026plusmn;0.003\u003csup\u003ei\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.244\u0026plusmn;0.001\u003csup\u003ei\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.581\u0026plusmn;0.001\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.674\u0026plusmn;0.002\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.086\u0026plusmn;0.001\u003csup\u003eg\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAscorbic acid\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e250 ppm\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.713\u0026plusmn;0.007\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.473\u0026plusmn;0.005\u003csup\u003en\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.222\u0026plusmn;0.002\u003csup\u003en\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.443\u0026plusmn;0.001\u003csup\u003eo\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.880\u0026plusmn;0.001\u003csup\u003ep\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.842\u0026plusmn;0.001\u003csup\u003ep\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.743\u0026plusmn;0.007\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.102\u0026plusmn;0.003\u003csup\u003el\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.185\u0026plusmn;0.004\u003csup\u003el\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.205\u0026plusmn;0.003\u003csup\u003en\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.827\u0026plusmn;0.006\u003csup\u003eo\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.319\u0026plusmn;0.004\u003csup\u003eo\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e150\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.438\u0026plusmn;0.002\u003csup\u003eg\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.874\u0026plusmn;0.002\u003csup\u003ef\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.787\u0026plusmn;0.002\u003csup\u003ef\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.768\u0026plusmn;0.001\u003csup\u003ej\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.143\u0026plusmn;0.002\u003csup\u003ek\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.007\u0026plusmn;0.002\u003csup\u003el\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e200\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.716\u0026plusmn;0.004\u003csup\u003eh\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.969\u0026plusmn;0.002\u003csup\u003ej\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.567\u0026plusmn;0.001\u003csup\u003ej\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.725\u0026plusmn;0.001\u003csup\u003eh\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.945\u0026plusmn;0.001\u003csup\u003ei\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.745\u0026plusmn;0.002\u003csup\u003ek\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Discussion","content":"\u003cp\u003ePlants under salt stress have unique physiological and biochemical alterations that impact them from the beginning of the stress to maturity (Munns \u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Significant decreases in growth metrics, such as root and shoot length, fresh and dry biomass, and leaf area, were seen in plants cultivated under different salt levels in this study. These consequences are linked to salt stress-induced cell shrinkage and dehydration, which limits cell elongation and division and, in turn, lowers the rate at which roots, shoots, and leaves grow. Osmotic alterations outside the roots are linked to these reactions, which hinder water intake and interfere with cell-water connections (Munns \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Our data support the conclusions of (Ghoulam et al. 2002), that demonstrated a discernible decrease in growth features due to salt, and with the investigation of (Kaya et al. 2018). Combining selenium, α-tocopherol, and ascorbic acid increased the growth of maize plants under salt stress. By strengthening antioxidant defences and regulating the metabolism of secondary metabolites, selenium in particular helps to regulate development (Jiang et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Ascorbic acid helps plants grow and become more resilient to stress, in part by facilitating the production of proteins that are high in hydroxyproline (Alamri et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Because chloroplasts are extremely sensitive to salinity, large salt concentrations can destabilise pigment\u0026ndash;protein complexes and cause structural damage. Under salt stress, carotenoids suffer as well, and the amount of chlorophyll drops because of an increase in chlorophyllase activity (Mosavi et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Changes in nitrogen metabolism, including increased synthesis of proline, which is essential for osmotic regulation, may be connected to the decrease in chlorophyll content. Glutamate kinase activity, the primary enzyme that starts proline production, is further stimulated by salt stress. (Shahbaz et al. \u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; And and Dawood 2014). Research by (El-Sawy \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2009\u003c/span\u003e and Hassanein et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) This suggests that decreased uptake of essential ions such as Mg\u0026sup2;⁺ and Fe\u0026sup3;⁺, which are essential for chlorophyll biosynthesis under stress, may be linked to the decline in plant pigment levels. Salinity also dramatically lowers stomatal conductance, photosynthetic electron transport efficiency, and carbon assimilation (Sun et al. \u003cspan citationid=\"CR118\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The buildup of Na\u003csup\u003e+\u003c/sup\u003e and Cl\u003csup\u003e+\u003c/sup\u003e ions in plant tissues is a major sign of salt-induced stress, which upsets physiological equilibrium by changing the uptake of vital soil elements (Carvalho et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). In both stress and non-stress situations, enough selenium improves CO₂ assimilation and chlorophyll fluorescence, which increases photosynthetic efficiency (Alyemeni et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Additionally, applying selenium to stressed plants promoted growth and raised the amount of photosynthetic pigment in Melissa officinalis L. (Habibi \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), In contrast, Selenium raised K\u003csup\u003e+\u003c/sup\u003e and reduced Na\u003csup\u003e+\u003c/sup\u003e buildup in the roots and shoots. (Shekari, Abbasi, and Mustafavi 2017). A-tocopherol administered exogenously was discovered by (Das and Roychoudhury 2014) to significantly increase photosynthetic pigment concentrations. Plant leaves' photosynthetic membranes contain a-tocopherol (Grilo et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Tocopherols are important for chloroplast heat dissipation, scavenge reactive oxygen species (ROS), and prevent lipid peroxidation. They are found in large quantities in cellular membranes (Falk and Munn\u0026eacute;-Bosch 2010). According to (Kruk and Strzalka 2001), In photosystem II, α-tocopherol protects chloroplasts by reducing cytochrome b\u003csub\u003e559\u003c/sub\u003e activity during cyclic electron flow. As mentioned by (Semida et al. \u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), Consistent with observations observed in other crops, exogenous α-tocopherol increased Antioxidant actions, both enzymatic and non-enzymatic, preserved cellular turgor, and enhanced photosynthetic efficiency in onion plants under salt stress. Solute loss, also known as electrolyte leakage, from plant cells can be measured to indirectly measure membrane damage (Ekmek\u0026ccedil;i et al. 2008), and assessing the membrane stability index (Ali and Abbas 2003). The findings unequivocally demonstrate that, in comparison to controls, high salinity considerably decreased the membrane stability index and increased electrolyte leakage in maize plants Jamil et al. (\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), It was shown that larger amounts of salt, in particular, resulted in more damage to cell membranes. (Hniličkov\u0026aacute; et al. 2019) Higher salt concentrations were shown to enhance electrolyte leakage in all species. Leakage in Lactuca sativa reached 27.5%, indicating a significant difference between the control and 50 mmol/L NaCl treatments. Additionally, (Mahmoudi et al. \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) With rising NaCl concentrations, electrolyte leakage in lettuce roots and leaves was seen to increase gradually. As a membrane-bound antioxidant and scavenger of reactive oxygen species (ROS), α-tocopherol assisted in lowering lipid peroxidation in maize plants (Orabi and Abdelhamid 2016). Exogenous α-tocopherol increased antioxidant enzyme activity and decreased MDA levels, which lessened the negative effects of seawater salinity on faba beans ( Semida et al. \u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). As previously reported, ascorbic acid application successfully decreased membrane lipid peroxidation in maize, most likely by strengthening the plant's antioxidant defence mechanism (Ahmad et al. 2016; Alyemeni et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Supplementing with selenium has been shown to increase the uptake of nitrogen, potassium, and calcium, as well as amino acid synthesis and stress signalling, all of which help wheat become more resistant to salt (Elkelish et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). When selenium-amino acids were added to proteins, their biological activity changed, encouraging cucumber seedlings under salt stress to produce more proline (Hasanuzzaman et al. 2011). When α-tocopherol was applied exogenously to maize, it improved nitrogen metabolism, which led to increased proline buildup, better growth, and more yield components (Buschmann and Lichtenthaler \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1979\u003c/span\u003e). Applying ascorbic acid to maize increased its protein content and nitrogen metabolism, according to (El-Bassiouny and Sadak 2015). Moreover, ascorbic acid and α-tocopherol applied topically significantly raised the amounts of protein and carbohydrates in flax cultivars under salt stress (Sadak and Dawood 2014).\u003c/p\u003e \u003cp\u003eCarbohydrates, phenolics, and flavonoids are frequently linked to active osmotic adjustment, as noted by (Stepien and Klobus \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). The amount of total carbohydrates, phenolics, and flavonoids in maize plants decreased gradually as the salinity level increased under NaCl. Growth reductions were reflected in changes in carbohydrate content, which connected lower photosynthetic production to lower shoot dry weight. In salt-stressed maize, the decrease in chlorophyll and carbohydrates suggests either decreased photosynthesis or higher diversion of carbohydrates to other metabolic pathways (Handa et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Shaddad et al. \u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). Abiotic and biotic stressors, such as salinity, often affect the synthesis of phenolics (Parida et al. \u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Decreased activity of the essential enzyme that initiates the phenylpropanoid pathway, phenylalanine ammonia-lyase (PAL), which is necessary for plants to produce polyphenolic chemicals such flavonoids, phenylpropanoids, and lignin, is thought to be the cause of the drop in phenolic synthesis under stress (FRITZ et al. 1976; Tanaka et al. \u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). Furthermore, the absorption of potassium and phosphorus, which are necessary for secondary metabolites like polyphenols, is decreased at high salt concentrations (Rezazadeh et al. \u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). However, some research found that plants treated with salt had fewer phenolic chemicals (Wahid and Ghazanfar 2006; Blasco et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), others indicated enhanced phenolic biosynthesis (Burchard et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Mahmoudi et al. \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). The negative effects of salinity were partially mitigated by the individual administrations of Na₂SeO₄, α-tocopherol, and ascorbic acid, which usually improved the accumulation of carbohydrates, phenolics, and flavonoids in salt-stressed maize. Because of their absorption and storage in the green tissues, supplements containing sodium selenate or α-tocopherol under salt stress markedly increased physico-biochemical features, such as proteins, carbohydrates, carotenoids, and chlorophyll. In Melissa officinalis L. under Na\u003csup\u003e+\u003c/sup\u003e stress, selenium treatment also had a favourable impact on growth, photosynthetic pigments, and total carbohydrate content (Habibi \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). α-tocopherol's antioxidant qualities were essential in reducing the negative effects of ROS brought on by salt stress (Attallah et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). By stabilising biological membranes and limiting lipid peroxidation, α-tocopherol in plastid membranes protected chloroplasts, the main location for sugar production in plants. (Wang and Quinn 2000; Sharma et al. \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Spicher, Glauser, and Kessler 2016). By increasing photosynthesis, boosting nutrient uptake, and decreasing the buildup of Na⁺ in tissues, exogenous ascorbic acid enabled maize plants treated with Na⁺ withstand oxidative damage. Likewise, under drought stress, seed priming with ascorbic acid raised Vicia faba's leaf chlorophyll, hydration status, and total carbohydrate content (Azooz et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Ascorbic acid stimulated transpiration, photosynthesis, plant development, and the amount of carbohydrates (Naz et al. 2016; Alamri et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Ibrahim et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) showed that when exposed to 5000 ppm salinity, the phenolic and flavonoid levels rose and interacted with 150 ppm ascorbic acid. (Wu et al. \u003cspan citationid=\"CR128\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), after being treated with ascorbic acid, Geranium sibiricum L. also showed an increase in total phenolic content. In addition to increased phenylalanine ammonia-lyase (PAL) and catalase (CAT) activities during the study period under constant NaCl levels, salt stress enhances the activities of amylases, proteases, polyphenol oxidase, and peroxidase. Salinity has been shown to increase amylase and protease activities in the cotyledons of sprouting cotton seeds, but it also delays reserve mobilisation, which is linked to decreased activity of reserve-degrading enzymes. (Ashraf et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2002\u003c/span\u003e), whereas during late seedling establishment it reduces the activities of lipases, amylases, and proteases in cashew cotyledons (Marques et al. 2013). (Zeid et al. \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), Optimal quantities of ascorbic acid, gibberellic acid, sodium selenate (Na₂SeO₄), or selenium nanoparticles (SeNPs) enhanced the activities of α-amylase, β-amylase, and protease in seedlings, but higher concentrations resulted in a decrease. When α-tocopherol was applied to maize under water stress, it reduced amylase activity and encouraged the buildup of carbohydrates (Terzi et al. 2018). (Abdallah et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) The activities of α-amylase, β-amylase, and protease in seedlings were increased by optimal amounts of ascorbic acid, gibberellic acid, sodium selenate (Na₂SeO₄), or selenium nanoparticles (SeNPs); however, higher concentrations caused a decrease. α-tocopherol decreased amylase activity and promoted the accumulation of carbohydrates in maize under water stress (Hammerschmidt et al. 1982; Fukasawa-Akada et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Thipyapong and Steffens 1997). In order to reduce the excessive formation of reactive oxygen species (ROS) brought on by salt, selenium is essential for both enzymatic processes like catalase (CAT) and peroxidase (POD) activities as well as non-enzymatic mechanisms involving phytochelatins and glutathione (Zong et al. \u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). It has been said that applying Se is crucial for boosting ROS scavenging activity and for naturally boosting plant antioxidant activity (Shalaby et al. \u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Additionally, it has been demonstrated that supplementing with Se reduces the production of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e during salt stress, as seen by improvements in the antioxidant enzyme activities (POD and CAT) in canola and dill seedlings (Hasanuzzaman et al. 2011; Shekari et al. 2017). Exogenous α-tocopherol's beneficial effects on a range of physio-biochemical characteristics most likely result from its involvement in several physiological functions and its internal translocation inside the plant, which improves overall stress tolerance (Kumar et al. \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). As a cofactor for many antioxidant enzymes, ascorbic acid supports a number of physiological functions, such as controlling the amount of proteins and nucleic acids, membrane permeability, photosynthesis, root ion uptake, respiration, seed germination, and enzyme activity in a variety of plant species (Tavili et al. \u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Ozdener and Kutbay 2008; Ejaz et al. 2012). (Farooq et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) showed that CAT and POD enzyme activities rose in response to water stress, which is in line with previous findings of increased PPO, CAT, and POD enzyme activities in wheat treated with ascorbic acid (Shafiq et al. \u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Hafez and Gharib \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eProtein profiles change as a result of salinity stress, and stress-specific proteins that shield cells from salt damage are also induced. In control plants, vitamins have been demonstrated to alter gene expression, increasing the synthesis of preexisting proteins and encouraging the emergence of new protein bands (Bassuony et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Al-Hafedh et al. 2008). Vitamins have a substantial impact on the amount and quality of protein composition, which may improve growth and productivity, as evidenced by the observed increase in the intensity of original protein bands in control plants. Furthermore, one important defence against salt stress is the de novo creation of particular proteins (Hassanein et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Eid \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Many biosynthetic and signalling pathways control the genetically complicated trait of salt tolerance in plants. Numerous investigations have documented changes in polypeptide patterns in response to salt stress, both qualitatively and quantitatively (Chawla et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Arefian et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Baniulis et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Salinity decreased the protein content of pigeon peas (Cajanus cajan), with the decline being more noticeable at 100 mM NaCl than at 50 mM NaCl (Chavan et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Through alterations in one-dimensional SDS-PAGE profiles, the current work shows that gene expression in maize is modulated by both NaCl and individual administrations of sodium selenate, α-tocopherol, and ascorbic acid. The seeds treated with α-tocopherol produced two bands at 59.296 and 39.285 kDa, ascorbic acid produced bands at 80.472 and 67.439 kDa, and presoaking in sodium selenate produced a novel protein band at 82.846 kDa. These recently produced proteins most likely serve as osmoprotectants, metabolic enzymes, ROS scavengers, signal transducers, or are involved in the biogenesis, degradation, and defence processes of cells (Sairam and Tyagi 2004; (Chattopadhyay et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Lisa et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Kappel et al. \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). While SDS-PAGE cannot definitively identify proteins, the observed patterns indicate that selenium, α-tocopherol, and ascorbic acid modulate the expression of constitutive and stress-inducible proteins. Bands in the 95\u0026ndash;72 kDa range may correspond to Rubisco or other photosynthetic proteins, 67\u0026ndash;59 kDa to stress-related enzymes, and lower molecular weight bands (44\u0026ndash;34 kDa) to regulatory or defense proteins, highlighting the physiological adjustments of maize under abiotic stress and antioxidant treatments.\u003c/p\u003e \u003cp\u003eExcessive Na\u003csup\u003e+\u003c/sup\u003e buildup in plants interferes with stomatal control by upsetting K\u003csup\u003e+\u003c/sup\u003e equilibrium. Therefore, maintaining a high K⁺/Na⁺ ratio is crucial for salt tolerance (Shabala et al. \u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Abbasi et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Crops adjust to salinity by limiting the movement of Na\u003csup\u003e+\u003c/sup\u003e and Cl\u003csup\u003e+\u003c/sup\u003e ions to stems and leaf sheaths, which reduces their buildup in more vulnerable leaf blades (Isla and Arag\u0026uuml;\u0026eacute;s 2010). Sustaining K\u003csup\u003e+\u003c/sup\u003e intake, decreasing K\u003csup\u003e+\u003c/sup\u003e efflux, restricting Na\u003csup\u003e+\u003c/sup\u003e influx, and encouraging Na\u003csup\u003e+\u003c/sup\u003e efflux from cells are all necessary to maintain the cytosolic K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e ratio (Wakeel et al. \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). It has been demonstrated that high concentrations of NaCl in the growth medium decrease K\u003csup\u003e+\u003c/sup\u003e ion absorption (Faheed \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Noreen et al. \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Relative water content is decreased when hydraulic conductivity is decreased due to salinity-induced Na\u003csup\u003e+\u003c/sup\u003e buildup in roots. On the other hand, supplementing with selenium (Na₂SeO₄) reduces Na⁺ toxicity, encourages root growth, and improves water transfer to shoots, all of which boost overall plant growth (Rietz and Haynes \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Because proline production is necessary for preserving protein stability, saline soils interfere with nitrogen uptake, accumulation, and metabolism (Carillo et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Szabados and Savoure 2010). It has been demonstrated that selenium enhances nutrient uptake and translocation in a variety of crops, boosting yield and growth (Shahzadi et al. \u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Interestingly, when compared to untreated plants, selenite-treated plants showed greater K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e ratios and lower Na\u003csup\u003e+\u003c/sup\u003e concentrations (Subramanyam et al., 2019). Applying selenium has been associated with a decrease in Na\u003csup\u003e+\u003c/sup\u003e buildup, which raises the K\u003csup\u003e+\u003c/sup\u003e/Na\u003csup\u003e+\u003c/sup\u003e ratio in plants that produce garlic (Allium sativum L.) and dill (Anethum graveolens) (Shekari, Abbasi, and Mustafavi 2017); (Astaneh et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In a similar vein, applying selenium to Melissa officinalis L. improved growth, photosynthetic pigments, and total amino acid content while decreasing Na\u003csup\u003e+\u003c/sup\u003e and increasing K\u003csup\u003e+\u003c/sup\u003e levels in Anethum graveolens plants (Habibi \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2017\u003c/span\u003e;Shekari et al. 2017). Antioxidants like α-tocopherol, which improve osmotolerance and control processes like membrane permeability and nutrient uptake from the soil, are responsible for the increased nutritional content (Orabi and Abdelhamid 2016). Applying α-tocopherol has been associated with increased leaf ion content, which improves nutrient uptake and has a beneficial effect on plant growth (Buschmann and Lichtenthaler \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1979\u003c/span\u003e); (Sadak and Dawood 2014). Additionally, α-tocopherol has been shown to be successful in reducing the harmful effects of saltwater salinity on faba beans (Orabi and Abdelhamid 2016). Applying α-tocopherol to flax cultivars has been demonstrated to improve plant growth and yield by raising Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e, Mg\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e, and K\u003csup\u003e+\u003c/sup\u003e levels while decreasing Na\u003csup\u003e+\u003c/sup\u003e and Cl\u003csup\u003e\u0026minus;\u003c/sup\u003e levels (Sadak et al. \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Semida et al. \u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). While K⁺ content in both tissues dropped as NaCl concentration increased, ascorbic acid treatment dramatically inhibited Na⁺ buildup in faba bean roots and shoots (Loutfy et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Additionally, under salt stress conditions, ascorbic acid has been shown to increase the levels of K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e, and Mg\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e while decreasing the buildup of Na\u003csup\u003e+\u003c/sup\u003e (El-Bassiouny and Sadak 2015).\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003e According to the present study, salt-induced oxidative stress markedly inhibits plant growth and development by impairing photosynthetic efficiency, disrupting nutrient homeostasis, and elevating multiple stress biomarkers in a concentration-dependent manner. In contrast, the exogenous application of selenium, α-tocopherol, and ascorbic acid significantly improves growth performance in salt-stressed plants. This protective treatment enhances chlorophyll biosynthesis and stimulates the accumulation of secondary metabolites. Notably, ascorbic acid proved to be more effective than selenium and α-tocopherol in alleviating salt stress. Its superior efficacy is attributed to efficient scavenging of reactive oxygen species (ROS), enhanced osmolyte (proline) accumulation, and reinforcement of the antioxidant defense system. Consequently, ascorbic acid improves plant water relations, growth attributes, and photosynthetic efficiency under saline conditions. Furthermore, this treatment induces the expression of proteins associated with salt-stress tolerance. Collectively, these findings highlight a promising, eco-friendly, and cost-effective strategy for mitigating salt toxicity in agricultural systems worldwide.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eDeclaration of competing interest\u003c/h2\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003ch2\u003eFunding\u003c/h2\u003e\n\u003cp\u003eThis research was funded by Prince Sattam bin Abdulaziz University under project number PSAU/2025/01/32873\u003c/p\u003e\n\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eRK and MY\u0026ndash; Conceive and conceptualize the idea; FB and SH\u0026ndash; Investigated and collected data; RK, MY, AG \u0026ndash; Analyzed data and prepared graphs; MY and RK\u0026ndash; Prepared original draft and revised, MY\u0026ndash; Edited and content improvement, MR, HN, MA, MR, YD \u0026ndash; Statistical analysis and revised the draft.\u003c/p\u003e\n\u003ch2\u003eAcknowledgments\u003c/h2\u003e\n\u003cp\u003eThe authors would like to thank the Botany and microbiology department, Faculty of Science, Benha University. For permitting us to carry out the experiment using their laboratory facilities. Also, the authors extend their appreciation to prince Sattam bin Abdulaziz University for funding this research work through the project number (PSAU/2025/01/32873).\u003c/p\u003e\n\u003ch2\u003eAvailability of data and material (data transparency)\u003c/h2\u003e\n\u003cp\u003eThe data that support the findings of this study are available from Prince Sattam bin Abdulaziz University and Benha University but restrictions apply to the availability of these data, which were used under license for the current study, and so are not publicly available. Data are however available from the authors upon reasonable request and with permission of Prince Sattam bin Abdulaziz University and Benha University.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbbasi, Ghulam Hasan, Javaid Akhtar, Rafiq Ahmad, Moazzam Jamil, Muhammad Anwar-ul-Haq, Shafaqat Ali, and Muhammad Ijaz. 2015. \u0026ldquo;Potassium Application Mitigates Salt Stress Differentially at Different Growth Stages in Tolerant and Sensitive Maize Hybrids.\u0026rdquo; \u003cem\u003ePlant Growth Regulation\u003c/em\u003e 76 (1): 111\u0026ndash;25.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbdallah, M M, Abd El-Monem AA, R A Hassanein, and H M S El-Bassiouny. 2013. \u0026ldquo;Response of Sunflower Plant to the Application of Certain Vitamins and Arbuscular Mycorrhiza under Different Water Regimes.\u0026rdquo; \u003cem\u003eAust J Basic Appl Sci\u003c/em\u003e 7: 915\u0026ndash;32.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAebi, Hugo. 1984. \u0026ldquo;[13] Catalase in Vitro.\u0026rdquo; In \u003cem\u003eMethods in Enzymology\u003c/em\u003e, 105:121\u0026ndash;26. Elsevier.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAhmad, Parvaiz, E F Abd Allah, Abeer Hashem, Maryam Sarwat, and Salih Gucel. 2016. \u0026ldquo;Exogenous Application of Selenium Mitigates Cadmium Toxicity in Brassica Juncea L.(Czern \u0026amp; Cross) by up-Regulating Antioxidative System and Secondary Metabolites.\u0026rdquo; \u003cem\u003eJournal of Plant Growth Regulation\u003c/em\u003e 35 (4): 936\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-Hafedh, Yousef S, Aftab Alam, and Mohamed Salaheldin Beltagi. 2008. \u0026ldquo;Food Production and Water Conservation in a Recirculating Aquaponic System in Saudi Arabia at Different Ratios of Fish Feed to Plants.\u0026rdquo; \u003cem\u003eJournal of the World Aquaculture Society\u003c/em\u003e 39 (4): 510\u0026ndash;20.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlamri, Saud A, Manzer H Siddiqui, Mutahhar Y Y Al-khaishany, M Nasir, Hayssam M Ali, Ibrahim A Alaraidh, Abdulaziz A Alsahli, Hala Al-, and Mohammed Mateen. 2018. \u0026ldquo;Ascorbic Acid Improves the Tolerance of Wheat Plants to Lead Toxicity.\u0026rdquo; \u003cem\u003eJournal of Plant Interactions\u003c/em\u003e 9145 (1): 409\u0026ndash;19. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/17429145.2018.1491067\u003c/span\u003e\u003cspan address=\"10.1080/17429145.2018.1491067\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAli, Refaat Mohamed, and H. M. Abbas. 2003. \u0026ldquo;Response of Salt Stressed Barley Seedlings to Phenylurea.\u0026rdquo; \u003cem\u003ePlant, Soil and Environment\u003c/em\u003e 49 (4): 158\u0026ndash;62.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlyemeni, Mohammed Nasser, Mohammad Abass Ahanger, Leonard Wijaya, Pravej Alam, Renu Bhardwaj, and Parvaiz Ahmad. 2018. \u0026ldquo;Selenium Mitigates Cadmium-Induced Oxidative Stress in Tomato (Solanum Lycopersicum L.) Plants by Modulating Chlorophyll Fluorescence, Osmolyte Accumulation, and Antioxidant System.\u0026rdquo; \u003cem\u003eProtoplasma\u003c/em\u003e 255 (2): 459\u0026ndash;69.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAnd, Sadak, and Dawood. 2014. \u0026ldquo;Role of Ascorbic Acid and \u003cspan\u003e$\u003c/span\u003eα\u003cspan\u003e$\u003c/span\u003e Tocopherol in Alleviating Salinity Stress on Flax Plant (Linum Usitatissimum L.).\u0026rdquo; \u003cem\u003eJournal of Stress Physiology \u0026amp; Biochemistry\u003c/em\u003e 10 (1): 93\u0026ndash;111.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArefian, Mohammad, Saeedreza Vessal, Saeid Malekzadeh-Shafaroudi, Kadambot H M Siddique, and Abdolreza Bagheri. 2019. \u0026ldquo;Comparative Proteomics and Gene Expression Analyses Revealed Responsive Proteins and Mechanisms for Salt Tolerance in Chickpea Genotypes.\u0026rdquo; \u003cem\u003eBMC Plant Biology\u003c/em\u003e 19 (1): 300.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12870-019-1793-z\u003c/span\u003e\u003cspan address=\"10.1186/s12870-019-1793-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArif, Yamshi, Priyanka Singh, Husna Siddiqui, Andrzej Bajguz, and Shamsul Hayat. 2020. \u0026ldquo;Salinity Induced Physiological and Biochemical Changes in Plants: An Omic Approach towards Salt Stress Tolerance.\u0026rdquo; \u003cem\u003ePlant Physiology and Biochemistry\u003c/em\u003e 156: 64\u0026ndash;77.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAshraf, M Y, G Sarwar, M Ashraf, R Afaf, and A Sattar. 2002. \u0026ldquo;Salinity Induced Changes in \u003cspan\u003e$\u003c/span\u003eα\u003cspan\u003e$\u003c/span\u003e-Amylase Activity during Germination and Early Cotton Seedling Growth.\u0026rdquo; \u003cem\u003eBiologia Plantarum\u003c/em\u003e 45 (4): 589\u0026ndash;91.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAstaneh, Rozita Khademi, Sahebali Bolandnazar, Fariborz Zaare Nahandi, and Shahin Oustan. 2018. \u0026ldquo;The Effects of Selenium on Some Physiological Traits and K, Na Concentration of Garlic (Allium Sativum L.) under NaCl Stress.\u0026rdquo; \u003cem\u003eInformation Processing in Agriculture\u003c/em\u003e 5 (1): 156\u0026ndash;61.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAttallah, Abdelfattah M, Faisal A Bughdadi, Atef M El-Shazly, and Hisham Ismail. 2013. \u0026ldquo;Immunodetection of Fasciola Gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis.\u0026rdquo; \u003cem\u003eClinical and Vaccine Immunology\u003c/em\u003e 20 (10): 1569\u0026ndash;77.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAzooz, Mohamed M, Abdullah M Alzahrani, Magdy M Youssef, and others. 2013. \u0026ldquo;The Potential Role of Seed Priming with Ascorbic Acid and Nicotinamide and Their Interactions to Enhance Salt Tolerance in Broad Bean ('Vicia Faba\u0026rsquo;L.).\u0026rdquo; \u003cem\u003eAustralian Journal of Crop Science\u003c/em\u003e 7 (13): 2091\u0026ndash;2100.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaniulis, Danas, Monika Sirgediene, Perttu Haimi, Inga Tamošiune, and Darius Danusevičius. 2020. \u0026ldquo;Constitutive and Cold Acclimation-Regulated Protein Expression Profiles of Scots Pine Seedlings Reveal Potential for Adaptive Capacity of Geographically Distant Populations.\u0026rdquo; \u003cem\u003eForests\u003c/em\u003e 11 (1): 89.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBassuony, F M, R A Hassanein, D M Baraka, and R R Khalil. 2008. \u0026ldquo;Physiological Effects of Nicotinamide and Ascorbic Acid on Zea Mays Plant Grown under Salinity Stress. II-Changes in Nitrogen Constituents, Protein Profiles, Protease Enzyme and Certain Inorganic Cations.\u0026rdquo; \u003cem\u003eAustralian Journal of Basic and Applied Sciences\u003c/em\u003e 2 (3): 350\u0026ndash;59.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBates, L S, R P Waldren, and I D Teare. 1973. \u0026ldquo;Rapid Determination of Free Proline for Water-Stress Studies.\u0026rdquo; \u003cem\u003ePlant and Soil\u003c/em\u003e 39 (1): 205\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlasco, Begon?a, Rocio Leyva, Luis Romero, and Juan Manuel Ruiz. 2013. \u0026ldquo;Iodine Effects on Phenolic Metabolism in Lettuce Plants under Salt Stress.\u0026rdquo; \u003cem\u003eJournal of Agricultural and Food Chemistry\u003c/em\u003e 61 (11): 2591\u0026ndash;96.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBurchard, P, W Bilger, and G Weissenb\u0026ouml;ck. 2000. \u0026ldquo;Contribution of Hydroxycinnamates and Flavonoids to Epidermal Shielding of UV-A and UV-B Radiation in Developing Rye Primary Leaves as Assessed by Ultraviolet-Induced Chlorophyll Fluorescence Measurements.\u0026rdquo; \u003cem\u003ePlant, Cell \u0026amp; Environment\u003c/em\u003e 23 (12): 1373\u0026ndash;80.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuschmann, C, and H K Lichtenthaler. 1979. \u0026ldquo;The Influence of Phytohormones on Prenyl Lipid Composition and Photosynthetic Activities of Thylakoids.\u0026rdquo; \u003cem\u003eAdvances in Biochemistry and Physiology of Plant Lipids. Elsevier, Amserdam\u003c/em\u003e, 145\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarillo, Petronia, Gabriella Mastrolonardo, Francesco Nacca, and Amodio Fuggi. 2005. \u0026ldquo;Nitrate Reductase in Durum Wheat Seedlings as Affected by Nitrate Nutrition and Salinity.\u0026rdquo; \u003cem\u003eFunctional Plant Biology\u003c/em\u003e 32 (3): 209\u0026ndash;19.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarvalho, Marcia E.A., Paulo R.C. Castro, Marcin Kozak, and Ricardo A. Azevedo. 2020. \u0026ldquo;The Sweet Side of Misbalanced Nutrients in Cadmium-Stressed Plants.\u0026rdquo; \u003cem\u003eAnnals of Applied Biology\u003c/em\u003e 176 (3): 275\u0026ndash;84. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/aab.12569\u003c/span\u003e\u003cspan address=\"10.1111/aab.12569\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChattopadhyay, Arnab, Pratigya Subba, Aarti Pandey, Deepti Bhushan, Rajiv Kumar, Asis Datta, Subhra Chakraborty, and Niranjan Chakraborty. 2011. \u0026ldquo;Analysis of the Grasspea Proteome and Identification of Stress-Responsive Proteins upon Exposure to High Salinity, Low Temperature, and Abscisic Acid Treatment.\u0026rdquo; \u003cem\u003ePhytochemistry\u003c/em\u003e 72 (10): 1293\u0026ndash;1307.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChavan, Nilesh S, Prashant B Kardile, and Roshan A Diware. 2018. \u0026ldquo;Effect of Salt Stress on Protein Profile of Different Varieties of Cajanus Cajan Using SDS Page.\u0026rdquo; \u003cem\u003eJournal of Pharmacognosy and Phytochemistry\u003c/em\u003e 7 (6): 1258\u0026ndash;62.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChawla, S, S C Goyal, K S Datta, R Angrish, and others. 2010. \u0026ldquo;Protein Profile Changes under Salt-Boron Toxicity and Its Regulation by Hydrogen Peroxide and Glutathione in Pigeonpea (Cajanus Cajan L.).\u0026rdquo; \u003cem\u003eIndian Journal of Plant Physiology\u003c/em\u003e 15 (4): 378\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eClark, J M, and R L Switzer. 1977. \u0026ldquo;In Experimental Biochemistry 2nd Ed.W.H. Freeman \u0026amp;Company, San Francisco.\u0026rdquo; Article.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDas, Kaushik, and Aryadeep Roychoudhury. 2014. \u0026ldquo;Reactive Oxygen Species (ROS) and Response of Antioxidants as ROS-Scavengers during Environmental Stress in Plants.\u0026rdquo; \u003cem\u003eFrontiers in Environmental Science\u003c/em\u003e 2: 53.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEid, Manal Hassan. 2019. \u0026ldquo;Gene Expression and Molecular Differences in Two Wheat Genotypes under Salt and Drought Stresses.\u0026rdquo; \u003cem\u003eEgyptian Journal of Agronomy\u003c/em\u003e 41 (2): 93\u0026ndash;104.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEjaz, Batool, Zahoor Ahmad Sajid, and Faheem Aftab. 2012. \u0026ldquo;Effect of Exogenous Application of Ascorbic Acid on Antioxidant Enzyme Activities, Proline Contents, and Growth Parameters of Saccharum Spp. Hybrid Cv. HSF-240 under Salt Stress.\u0026rdquo; \u003cem\u003eTurkish Journal of Biology\u003c/em\u003e 36 (6): 630\u0026ndash;40.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEkmek\u0026ccedil;i, Yasemin, Deniz Tanyolac, and Beycan Ayhan. 2008. \u0026ldquo;Effects of Cadmium on Antioxidant Enzyme and Photosynthetic Activities in Leaves of Two Maize Cultivars.\u0026rdquo; \u003cem\u003eJournal of Plant Physiology\u003c/em\u003e 165 (6): 600\u0026ndash;611.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Bassiouny, Hala M.S., and Mervat Sh Sadak. 2015. \u0026ldquo;Impact of Foliar Application of Ascorbic Acid and Alpha-Tocopherol on Antioxidant Activity and Some Biochemical Aspects of Flax Cultivars under Salinity Stress.\u0026rdquo; \u003cem\u003eActa Biologica Colombiana\u003c/em\u003e 20 (2): 209\u0026ndash;22. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.15446/abc.v20n2.43868\u003c/span\u003e\u003cspan address=\"10.15446/abc.v20n2.43868\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Sawy, O E H. 2009. \u0026ldquo;Protective Effects of Polyamines on Wheat Plants Irrigated by Seawater.\u0026rdquo; Ph. D. Thesis, Fac. Sci., Mans. Univ., Egypt.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eElkelish, Amr A, Mona H Soliman, Haifa A Alhaithloul, and Mohamed A El-Esawi. 2019. \u0026ldquo;Selenium Protects Wheat Seedlings against Salt Stress-Mediated Oxidative Damage by up-Regulating Antioxidants and Osmolytes Metabolism.\u0026rdquo; \u003cem\u003ePlant Physiology and Biochemistry\u003c/em\u003e 137: 144\u0026ndash;53.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEltayeb, Amin Elsadig, Naoyoshi Kawano, Ghazi Hamid Badawi, Hironori Kaminaka, Takeshi Sanekata, Toshiyuki Shibahara, Shinobu Inanaga, and Kiyoshi Tanaka. 2007. \u0026ldquo;Overexpression of Monodehydroascorbate Reductase in Transgenic Tobacco Confers Enhanced Tolerance to Ozone, Salt and Polyethylene Glycol Stresses.\u0026rdquo; \u003cem\u003ePlanta\u003c/em\u003e 225 (5): 1255\u0026ndash;64.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFadeel, A A. 1962. \u0026ldquo;Location and Properties of Chloroplasts and Pigment Determination in Roots.\u0026rdquo; \u003cem\u003ePhysiologia Plantarum\u003c/em\u003e 15 (1): 130\u0026ndash;46.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFaheed, Fayza A. 2012. \u0026ldquo;Comparative Effects of Four Herbicides on Physiological Aspects in Triticum Sativum L.\u0026rdquo; \u003cem\u003eAfrican Journal of Ecology\u003c/em\u003e 50 (1): 29\u0026ndash;42.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFalk, Jon, and Sergi Munn\u0026eacute;-Bosch. 2010. \u0026ldquo;Tocochromanol Functions in Plants: Antioxidation and Beyond.\u0026rdquo; \u003cem\u003eJournal of Experimental Botany\u003c/em\u003e 61 (6): 1549\u0026ndash;66.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFarooq, Ayesha, Shazia Anwer Bukhari, Nudrat A Akram, Muhammad Ashraf, Leonard Wijaya, Mohammed Nasser Alyemeni, and Parvaiz Ahmad. 2020. \u0026ldquo;Exogenously Applied Ascorbic Acid-Mediated Changes in Osmoprotection and Oxidative Defense System Enhanced Water Stress Tolerance in Different Cultivars of Safflower (Carthamus Tinctorious L.).\u0026rdquo; \u003cem\u003ePlants\u003c/em\u003e 9 (1): 104.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFRITZ, Taylor, Tatsuma ITO, Malek JAZIRI, John-Patrick SMITH, Alejandro GONZALEZ, Austin KRAJICEK, and Yoshihito NISHIOKA. 1976. \u0026ldquo;8 6 JPN.\u0026rdquo;\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFukasawa-Akada, Tomoko, Shain-dow Kung, and John C Watson. 1996. \u0026ldquo;Phenylalanine Ammonia-Lyase Gene Structure, Expression, and Evolution in Nicotiana.\u0026rdquo; \u003cem\u003ePlant Molecular Biology\u003c/em\u003e 30 (4): 711\u0026ndash;22.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhoulam, Cherki, Ahmed Foursy, and Khalid Fares. 2002. \u0026ldquo;Effects of Salt Stress on Growth, Inorganic Ions and Proline Accumulation in Relation to Osmotic Adjustment in Five Sugar Beet Cultivars.\u0026rdquo; \u003cem\u003eEnvironmental and Experimental Botany\u003c/em\u003e 47 (1): 39\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGong, D H, G Z Wang, W T Si, Y Zhou, Z Liu, and J Jia. 2018. \u0026ldquo;Effects of Salt Stress on Photosynthetic Pigments and Activity of Ribulose-1, 5-Bisphosphate Carboxylase/Oxygenase in Kalidium Foliatum.\u0026rdquo; \u003cem\u003eRussian Journal of Plant Physiology\u003c/em\u003e 65 (1): 98\u0026ndash;103.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGrilo, Evellyn C\u0026acirc;mara, Priscila Nunes Costa, Cristiane Santos S\u0026acirc;nzio Gurgel, Andressa Fernanda de Lima Beserra, Fernanda Ni\u0026eacute;ce de Souza Almeida, and Roberto Dimenstein. 2014. \u0026ldquo;Alpha-Tocopherol and Gamma-Tocopherol Concentration in Vegetable Oils.\u0026rdquo; \u003cem\u003eFood Science and Technology\u003c/em\u003e 34 (2): 379\u0026ndash;85.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHabibi, G. 2017. \u0026ldquo;Selenium Ameliorates Salinity Stress in Petroselinum Crispum by Modulation of Photosynthesis and by Reducing Shoot Na Accumulation.\u0026rdquo; \u003cem\u003eRussian Journal of Plant Physiology\u003c/em\u003e 64 (3): 368\u0026ndash;74.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHafez, E M, and H S Gharib. 2016. \u0026ldquo;Effect of Exogenous Application of Ascorbic Acid on Physiological and Biochemical Characteristics of Wheat under Water Stress.\u0026rdquo; \u003cem\u003eInt. J. Plant Prod\u003c/em\u003e 10 (4): 579\u0026ndash;96.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHammerschmidt, R, E M Nuckles, and J Kuć. 1982. \u0026ldquo;Association of Enhanced Peroxidase Activity with Induced Systemic Resistance of Cucumber to Colletotrichum Lagenarium.\u0026rdquo; \u003cem\u003ePhysiological Plant Pathology\u003c/em\u003e 20 (1): 73\u0026ndash;82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHanda, Sangita, Ray A Bressan, Avtar K Handa, Nicholas C Carpita, and Paul M Hasegawa. 1983. \u0026ldquo;Solutes Contributing to Osmotic Adjustment in Cultured Plant Cells Adapted to Water Stress.\u0026rdquo; \u003cem\u003ePlant Physiology\u003c/em\u003e 73 (3): 834\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHarris, D, A Rashid, G Miraj, M Arif, and H Shah. 2007. \u0026ldquo;On-Farmseed Priming with Zinc Sulphate SolutionA Cost-Effective Way to Increase the Maize Yields of Resource-Poor Farmers.\u0026rdquo; \u003cem\u003eField Crops Research\u003c/em\u003e 102 (2): 119\u0026ndash;27.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHarris, David, Abdul Rashid, Ghazal Miraj, Mohammed Arif, and Mohammed Yunas. 2008. \u0026ldquo;On-Farmseed Priming with Zinc in Chickpea and Wheat in Pakistan.\u0026rdquo; \u003cem\u003ePlant and Soil\u003c/em\u003e 306 (1\u0026ndash;2): 3\u0026ndash;10.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHartree, E Fo. 1972. \u0026ldquo;Determination of Protein: A Modification of the Lowry Method That Gives a Linear Photometric Response.\u0026rdquo; Article. \u003cem\u003eAnalytical Biochemistry\u003c/em\u003e 48 (2): 422\u0026ndash;27.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHasanuzzaman, Mirza, Mohammad Anwar Hossain, and Masayuki Fujita. 2011. \u0026ldquo;Selenium-Induced up-Regulation of the Antioxidant Defense and Methylglyoxal Detoxification System Reduces Salinity-Induced Damage in Rapeseed Seedlings.\u0026rdquo; \u003cem\u003eBiological Trace Element Research\u003c/em\u003e 143 (3): 1704\u0026ndash;21.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHassanein, R A, F M Bassuony, D M Baraka, and R R Khalil. 2009. \u0026ldquo;Physiological Effects of Nicotinamide and Ascrobic Acid on Zea Mays Plant Grown Under Salinity Stress. I-Changes in Growth, Some Relevant Metabolic Activities and Oxidative Defense Systems .\u0026rdquo; \u003cem\u003eJournal of Agriculture and Biological Sciences\u003c/em\u003e 5 (1): 72\u0026ndash;81.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHavir, Evelyn A, and Neil A McHale. 1987. \u0026ldquo;Biochemical and Developmental Characterization of Multiple Forms of Catalase in Tobacco Leaves.\u0026rdquo; \u003cem\u003ePlant Physiology\u003c/em\u003e 84 (2): 450\u0026ndash;55.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHniličkov\u0026aacute;, Helena, František Hnilička, Maty\u0026aacute;š Ors\u0026aacute;k, and V\u0026aacute;clav Hejn\u0026aacute;k. 2019. \u0026ldquo;Effect of Salt Stress on Growth, Electrolyte Leakage, Na\u0026thinsp;+\u0026thinsp;and K+ Content in Selected Plant Species.\u0026rdquo; \u003cem\u003ePlant, Soil and Environment\u003c/em\u003e 65 (2): 90\u0026ndash;96.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHodges, D Mark, John M DeLong, Charles F Forney, and Robert K Prange. 1999. \u0026ldquo;Improving the Thiobarbituric Acid-Reactive-Substances Assay for Estimating Lipid Peroxidation in Plant Tissues Containing Anthocyanin and Other Interfering Compounds.\u0026rdquo; \u003cem\u003ePlanta\u003c/em\u003e 207 (4): 604\u0026ndash;11.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIbrahim, Gamil E, Sami A Metwally, H Bedour, and M Sharbat. 2019. \u0026ldquo;Effect of Salinity and Ascorbic Acid on Geranium Volatile and Antioxidant Activity.\u0026rdquo; \u003cem\u003ePlant Archives\u003c/em\u003e 19 (2): 750\u0026ndash;57.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIsla, Ramon, and R\u0026aacute;mon Arag\u0026uuml;\u0026eacute;s. 2010. \u0026ldquo;Yield and Plant Ion Concentrations in Maize (Zea Mays L.) Subject to Diurnal and Nocturnal Saline Sprinkler Irrigations.\u0026rdquo; \u003cem\u003eField Crops Research\u003c/em\u003e 116 (1\u0026ndash;2): 175\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJamil, Muhammad, Muhammad Ashraf, S Rehman, Mushtaq Ahmad, and Eui Shik Rha. 2012. \u0026ldquo;Salinity Induced Changes in Cell Membrane Stability, Protein and RNA Contents.\u0026rdquo; \u003cem\u003eAfrican Journal of Biotechnology\u003c/em\u003e 11 (24): 6476\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJiang, Chaoqiang, Chaolong Zu, Dianjun Lu, Qingsong Zheng, Jia Shen, and Huoyan Wang. 2017. \u0026ldquo;Effect of Exogenous Selenium Supply on Photosynthesis, Na\u0026thinsp;+\u0026thinsp;Accumulation and Antioxidative Capacity of Maize (Zea Mays L .) under Salinity Stress.\u0026rdquo; \u003cem\u003eNature Publishing Group\u003c/em\u003e 7 (February): 1\u0026ndash;14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/srep42039\u003c/span\u003e\u003cspan address=\"10.1038/srep42039\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKappel, Lisa, Martin M\u0026uuml;nsterk\u0026ouml;tter, Gy\u0026ouml;rgy Sipos, Carolina Escobar Rodriguez, and Sabine Gruber. 2020. \u0026ldquo;Chitin and Chitosan Remodeling Defines Vegetative Development and Trichoderma Biocontrol.\u0026rdquo; \u003cem\u003ePLoS Pathogens\u003c/em\u003e 16 (2): e1008320.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKar, Manoranjan, and Dinabandhu Mishra. 1976. \u0026ldquo;Catalase, Peroxidase, and Polyphenoloxidase Activities during Rice Leaf Senescence.\u0026rdquo; \u003cem\u003ePlant Physiology\u003c/em\u003e 57 (2): 315\u0026ndash;19.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKaya, Cengiz, Muhammad Ashraf, and Osman Sonmez. 2018. \u0026ldquo;Combination of Nitric Oxide and Thiamin Regulates Oxidative Defense Machinery and Key Physiological Parameters in Salt-Stressed Plants of Two Maize Cultivars Differing in Salinity Tolerance.\u0026rdquo; \u003cem\u003eAdvances in Agricultural Science\u003c/em\u003e 6 (1): 34\u0026ndash;44. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://aaasjournal.org/submission/index.php/aaas/article/view/25\u003c/span\u003e\u003cspan address=\"http://aaasjournal.org/submission/index.php/aaas/article/view/25\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhalil, Radwan, Samia Haroun, Fardous Bassyoini, Aziza Nagah, and Mohammad Yusuf. 2021. \u0026ldquo;Salicylic Acid in Combination with Kinetin or Calcium Ameliorates Heavy Metal Stress in Phaseolus Vulgaris Plant.\u0026rdquo; \u003cem\u003eJournal of Agriculture and Food Research\u003c/em\u003e 5: 100182.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKlapheck, Sigrid, Ina Zimmer, and Heinz Cosse. 1990. \u0026ldquo;Scavenging of Hydrogen Peroxide in the Endosperm of Ricinus Communis by Ascorbate Peroxidase.\u0026rdquo; \u003cem\u003ePlant and Cell Physiology\u003c/em\u003e 31 (7): 1005\u0026ndash;13.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKruk, Jerzy, and Kazimierz Strzalka. 2001. \u0026ldquo;Redox Changes of Cytochrome b 559 in the Presence of Plastoquinones.\u0026rdquo; \u003cem\u003eJournal of Biological Chemistry\u003c/em\u003e 276 (1): 86\u0026ndash;91.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, Sanjeev, R Singh, and Harsh Nayyar. 2013. \u0026ldquo;{\u003cspan\u003e$\u003c/span\u003e}\\alpha{\\\u003cspan\u003e$\u003c/span\u003e}-Tocopherol Application Modulates the Response of Wheat (Triticum Aestivum L.) Seedlings to Elevated Temperatures by Mitigation of Stress Injury and Enhancement of Antioxidants.\u0026rdquo; \u003cem\u003eJournal of Plant Growth Regulation\u003c/em\u003e 32 (2): 307\u0026ndash;14.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLaemmli, Ulrich K. 1970. \u0026ldquo;Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4.\u0026rdquo; \u003cem\u003eNature\u003c/em\u003e 227 (5259): 680\u0026ndash;85.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, Yin, Zinan Wang, Xiaofen Sun, and Kexuan Tang. 2008. \u0026ldquo;Current Opinions on the Functions of Tocopherol Based on the Genetic Manipulation of Tocopherol Biosynthesis in Plants.\u0026rdquo; \u003cem\u003eJournal of Integrative Plant Biology\u003c/em\u003e 50 (9): 1057\u0026ndash;69.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, Zhao Hui, Qiang Wang, Xiao Ruan, Cun De Pan, and De An Jiang. 2010. \u0026ldquo;Phenolics and Plant Allelopathy.\u0026rdquo; \u003cem\u003eMolecules\u003c/em\u003e 15 (12): 8933\u0026ndash;52. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules15128933\u003c/span\u003e\u003cspan address=\"10.3390/molecules15128933\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLisa, Laisa A, Sabrina M Elias, M Sazzadur Rahman, Saima Shahid, Tetsushi Iwasaki, A K M Mahbub Hasan, Keiko Kosuge, Yasuo Fukami, and Zeba I Seraj. 2011. \u0026ldquo;Physiology and Gene Expression of the Rice Landrace Horkuch under Salt Stress.\u0026rdquo; \u003cem\u003eFunctional Plant Biology\u003c/em\u003e 38 (4): 282\u0026ndash;92.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLoutfy, Naglaa, Mohamed M Azooz, Ahmed M Hassanein, and Abdelkader Bassiony. 2019. \u0026ldquo;Potassium Synergize the Positive Effect of Ascorbic Acid on Some Morpho-Physiological Parameters of Salt Stressed Faba Bean Cultivars.\u0026rdquo; \u003cem\u003eEgyptian Journal of Botany\u003c/em\u003e 59 (3): 735\u0026ndash;51.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLowe, L E. 1993. \u0026ldquo;Water-Soluble Phenolic Materials.\u0026rdquo; \u003cem\u003eSoil Sampling and Methods of Analysis. Canadian Society of Soil Science. CRC Press, Boca Raton, FL, USA\u003c/em\u003e, 409\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMahmoudi, Hela, Jun Huang, Margaret Y Gruber, Rym Kaddour, Mokhtar Lachaal, Zeineb Ouerghi, and Abdelali Hannoufa. 2010. \u0026ldquo;The Impact of Genotype and Salinity on Physiological Function, Secondary Metabolite Accumulation, and Antioxidative Responses in Lettuce.\u0026rdquo; \u003cem\u003eJournal of Agricultural and Food Chemistry\u003c/em\u003e 58 (8): 5122\u0026ndash;30.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMahmoudi, Hela, Rym Kaddour, Jun Huang, Nawel Nasri, Ba\u0026acirc;tour Olfa, Sabah MRah, Abdelali Hannoufa, Mokhtar Lacha\u0026acirc;l, and Zeineb Ouerghi. 2011. \u0026ldquo;Varied Tolerance to NaCl Salinity Is Related to Biochemical Changes in Two Contrasting Lettuce Genotypes.\u0026rdquo; \u003cem\u003eActa Physiologiae Plantarum\u003c/em\u003e 33 (5): 1613\u0026ndash;22.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMalick, C P, and M B Singh. 1980. \u0026ldquo;Plant Enzymology and Histo Enzymology Kalyari\u0026rsquo;\u0026rsquo;Publishers New Delhi Pp, 286.\u0026rdquo;\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarques, Elton Camelo, Paulo Andr\u0026eacute; Ferreira de Freitas, Nara L\\\u0026rsquo;\\idia Mendes Alencar, Jos\u0026eacute; Tarquinio Prisco, and En\u0026eacute;as Gomes-Filho. 2013. \u0026ldquo;Increased Na\u0026thinsp;+\u0026thinsp;and Cl- Accumulation Induced by NaCl Salinity Inhibits Cotyledonary Reserve Mobilization and Alters the Source-Sink Relationship in Establishing Dwarf Cashew Seedlings.\u0026rdquo; \u003cem\u003eActa Physiologiae Plantarum\u003c/em\u003e 35 (7): 2171\u0026ndash;82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMosavi, Nosrat, Mostafa Ebadi, Mahdi Khorshidi, and Nahid Masoudian. 2018. \u0026ldquo;Study of Some Physiological Characteristics of Potato Tissue under Salinity Stress.\u0026rdquo; \u003cem\u003eInternational Journal of Farming and Allied Sciences\u003c/em\u003e 7 (1): 1\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMunn\u0026eacute;-Bosch, Sergi. 2005. \u0026ldquo;The Role of \u003cspan\u003e$\u003c/span\u003eα\u003cspan\u003e$\u003c/span\u003e-Tocopherol in Plant Stress Tolerance.\u0026rdquo; \u003cem\u003eJournal of Plant Physiology\u003c/em\u003e 162 (7): 743\u0026ndash;48.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e---. 2007. \u0026ldquo;{\u003cspan\u003e$\u003c/span\u003e}\\alpha{\\\u003cspan\u003e$\u003c/span\u003e}-Tocopherol: A Multifaceted Molecule in Plants.\u0026rdquo; \u003cem\u003eVitamins {\u0026amp;} Hormones\u003c/em\u003e 76: 375\u0026ndash;92.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMunn\u0026eacute;-Bosch, Sergi, and Leonor Alegre. 2002. \u0026ldquo;The Function of Tocopherols and Tocotrienols in Plants.\u0026rdquo; \u003cem\u003eCritical Reviews in Plant Sciences\u003c/em\u003e 21 (1): 31\u0026ndash;57.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMunns, Rana. 2002. \u0026ldquo;Comparative Physiology of Salt and Water Stress.\u0026rdquo; \u003cem\u003ePlant, Cell \u0026amp; Environment\u003c/em\u003e 25 (2): 239\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e---. 2005. \u0026ldquo;Genes and Salt Tolerance: Bringing Them Together.\u0026rdquo; \u003cem\u003eNew Phytologist\u003c/em\u003e 167 (3): 645\u0026ndash;63.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNaz, HIRA, Nudrat Aisha Akram, and Muhammad Ashraf. 2016. \u0026ldquo;Impact of Ascorbic Acid on Growth and Some Physiological Attributes of Cucumber (Cucumis Sativus) Plants under Water-Deficit Conditions.\u0026rdquo; \u003cem\u003ePak J Bot\u003c/em\u003e 48 (3): 877\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNguyen, Thi Bich Thuy, Saichol Ketsa, and Wouter G van Doorn. 2003. \u0026ldquo;Relationship between Browning and the Activities of Polyphenoloxidase and Phenylalanine Ammonia Lyase in Banana Peel during Low Temperature Storage.\u0026rdquo; \u003cem\u003ePostharvest Biology and Technology\u003c/em\u003e 30 (2): 187\u0026ndash;93.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNoreen, Sibgha, Ayesha Siddiq, Kousar Hussain, Shakeel Ahmad, and Mirza Hasanuzzaman. 2017. \u0026ldquo;Foliar Application of Salicylic Acid with Salinity Stress on Physiological and Biochemical Attributes of Sunflower (Helianthus Annuus L.) Crop.\u0026rdquo; \u003cem\u003eActa Scientiarum Polonorum-Hortorum Cultus\u003c/em\u003e 16 (2): 57\u0026ndash;74.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOrabi, Salwa A, and Magdi T Abdelhamid. 2016. \u0026ldquo;Protective Role of {\u003cspan\u003e$\u003c/span\u003e}\\alpha{\\\u003cspan\u003e$\u003c/span\u003e}-Tocopherol on Two Vicia Faba Cultivars against Seawater-Induced Lipid Peroxidation by Enhancing Capacity of Anti-Oxidative System.\u0026rdquo; \u003cem\u003eJournal of the Saudi Society of Agricultural Sciences\u003c/em\u003e 15 (2): 145\u0026ndash;54.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOzdener, Yasemin, and H Guray Kutbay. 2008. \u0026ldquo;Effect of Salinity and Temperature on the Germination of Spergularia Marina Seeds and Ameliorating Effect of Ascorbic and Salicylic Acids.\u0026rdquo; \u003cem\u003eJournal of Environmental Biology\u003c/em\u003e 29 (6): 959\u0026ndash;64.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eParida, Asish Kumar, Anath Bandhu Das, Yukika Sanada, and Prasanna Mohanty. 2004. \u0026ldquo;Effects of Salinity on Biochemical Components of the Mangrove, Aegiceras Corniculatum.\u0026rdquo; \u003cem\u003eAquatic Botany\u003c/em\u003e 80 (2): 77\u0026ndash;87.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRautenkranz, Andreas A F, Liantje Li, Felix Machler, Enrico Martinoia, and Johann J Oertli. 1994. \u0026ldquo;Transport of Ascorbic and Dehydroascorbic Acids across Protoplast and Vacuole Membranes Isolated from Barley (Hordeum Vulgare L. Cv Gerbel) Leaves.\u0026rdquo; \u003cem\u003ePlant Physiology\u003c/em\u003e 106 (1): 187\u0026ndash;93.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRezazadeh, Amir, Azim Ghasemnezhad, Mojtaba Barani, and Telmah Telmadarrehei. 2012. \u0026ldquo;Effect of Salinity on Phenolic Composition and Antioxidant Activity of Artichoke (Cynara Scolymus L.) Leaves.\u0026rdquo; \u003cem\u003eRes J Med Plant\u003c/em\u003e 6: 245\u0026ndash;52.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRietz, D N, and R J Haynes. 2003. \u0026ldquo;Effects of Irrigation-Induced Salinity and Sodicity on Soil Microbial Activity.\u0026rdquo; \u003cem\u003eSoil Biology and Biochemistry\u003c/em\u003e 35 (6): 845\u0026ndash;54.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRostami, Mahdi, and Hossein Abbaspour. 2019. \u0026ldquo;EFFECT OF SELENIUM ON GROWTH AND PHYSIOLOGICAL TRAITS OF BASIL PLANT (Ocimum Basilicum L.) UNDER ARSENIC STRESS CONDITIONS.\u0026rdquo; \u003cem\u003eREVISTA DE AGRICULTURA NEOTROPICAL\u003c/em\u003e 6 (3): 30\u0026ndash;37. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.32404/rean.v6i3.3324\u003c/span\u003e\u003cspan address=\"10.32404/rean.v6i3.3324\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSadak, and Dawood. 2014. \u0026ldquo;Role of Ascorbic Acid and {\u003cspan\u003e$\u003c/span\u003e}\\alpha{\\\u003cspan\u003e$\u003c/span\u003e}tocopherol in Alleviating Salinity Stress on Flax Plant (Linum Usitatissimum L.).\u0026rdquo; \u003cem\u003eJournal of Stress Physiology {\u0026amp;} Biochemistry\u003c/em\u003e 10 (1): 93\u0026ndash;111.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSadak, Mervat Sh, Asmaa R Abd El-Hameid, Faten S A Zaki, Mona G Dawood, and Mohamed E El-Awadi. 2020. \u0026ldquo;Physiological and Biochemical Responses of Soybean (Glycine Max L.) to Cysteine Application under Sea Salt Stress.\u0026rdquo; \u003cem\u003eBulletin of the National Research Centre\u003c/em\u003e 44 (1): 1\u0026ndash;10.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSadiq, Muhammad, NUDRAT AISHA Akram, and Muhammad Ashraf. 2017. \u0026ldquo;Foliar Applications of Alpha-Tocopherol Improves the Composition of Fresh Pods of Vigna Radiata Subjected to Water Deficiency.\u0026rdquo; \u003cem\u003eTurkish Journal of Botany\u003c/em\u003e 41 (3): 244\u0026ndash;52.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSairam, R K. 1994. \u0026ldquo;Effects of Homobrassinolide Application on Plant Metabolism and Grain Yield under Irrigated and Moisture-Stress Conditions of Two Wheat Varieties.\u0026rdquo; \u003cem\u003ePlant Growth Regulation\u003c/em\u003e 14 (2): 173\u0026ndash;81.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSairam, R K, and Aruna Tyagi. 2004. \u0026ldquo;Physiology and Molecular Biology of Salinity Stress Tolerance in Plants.\u0026rdquo; \u003cem\u003eCurrent Science\u003c/em\u003e, 407\u0026ndash;21.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSakanaka, Senji, Yumi Tachibana, and Yuki Okada. 2005. \u0026ldquo;Preparation and Antioxidant Properties of Extracts of Japanese Persimmon Leaf Tea (Kakinoha-Cha).\u0026rdquo; \u003cem\u003eFood Chemistry\u003c/em\u003e 89 (4): 569\u0026ndash;75.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSauvesty, A, F Page, and J Huot. 1992. \u0026ldquo;A Simple Method for Extracting Plant Phenolic Compounds.\u0026rdquo; \u003cem\u003eCanadian Journal of Forest Research\u003c/em\u003e 22 (5): 654\u0026ndash;59.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSemida, W M, R S Taha, M T Abdelhamid, and M M Rady. 2014. \u0026ldquo;Foliar-Applied α-Tocopherol Enhances Salt-Tolerance in Vicia Faba L. Plants Grown under Saline Conditions.\u0026rdquo; \u003cem\u003eSouth African Journal of Botany\u003c/em\u003e 95: 24\u0026ndash;31.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSemida, Wael M, Taia A Abd El-Mageed, Saad M Howladar, Mostafa M Rady, and others. 2016. \u0026ldquo;Foliar-Applied Alpha-Tocopherol Enhances Salt-Tolerance in Onion Plants by Improving Antioxidant Defence System.\u0026rdquo; \u003cem\u003eAustralian Journal of Crop Science\u003c/em\u003e 10 (7): 1030\u0026ndash;39.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSestak, Zdenek, J Catsky, and Paul Gordon Jarvis. 1971. \u0026ldquo;Determination of Chlorophyll a and b.In Z Sestak, J Catsky, PG Jarvis, Eds,.\u0026rdquo; \u003cem\u003ePlant Photosynthetic Production. Manual of Methods.The Hague, Netherlands, Dr. W. Junk NV, Pp\u003c/em\u003e, 672\u0026ndash;97.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShabala, Lana, Tracey A Cuin, Ian A Newman, and Sergey Shabala. 2005. \u0026ldquo;Salinity-Induced Ion Flux Patterns from the Excised Roots of Arabidopsis Sos Mutants.\u0026rdquo; \u003cem\u003ePlanta\u003c/em\u003e 222 (6): 1041\u0026ndash;50.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShaddad, M A, A F Radi, A M Abdel-Rahman, and M M Azooz. 1990. \u0026ldquo;Response of Seeds of Lupinus Termis and Vicia Faba to the Interactive Effect of Salinity and Ascorbic Acid or Pyridoxine.\u0026rdquo; \u003cem\u003ePlant and Soil\u003c/em\u003e 122 (2): 177\u0026ndash;83.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShafiq, Sidra, Nudrat Aisha Akram, Muhammad Ashraf, and Amara Arshad. 2014. \u0026ldquo;Synergistic Effects of Drought and Ascorbic Acid on Growth, Mineral Nutrients and Oxidative Defense System in Canola (Brassica Napus L.) Plants.\u0026rdquo; \u003cem\u003eActa Physiologiae Plantarum\u003c/em\u003e 36 (6): 1539\u0026ndash;53.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShahbaz, Muhammad, Noor Fatima, Zia Ur Rehman Mashwani, Abida Akram, Ehsan ul Haq, Asma Mehak, Fozia Abasi, et al. 2022. \u0026ldquo;Effect of Phytosynthesized Selenium and Cerium Oxide Nanoparticles on Wheat (Triticum Aestivum L.) against Stripe Rust Disease.\u0026rdquo; \u003cem\u003eMolecules\u003c/em\u003e 27 (23). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules27238149\u003c/span\u003e\u003cspan address=\"10.3390/molecules27238149\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShahzadi, Iram, Muhammad Iqbal, Rizwan Rasheed, M Arslan Ashraf, Shagufta Perveen, and Makhdoom Hussain. 2017. \u0026ldquo;Foliar Application of Selenium Increases Fertility and Grain Yield in Bread Wheat under Contrasting Water Availability Regimes.\u0026rdquo; \u003cem\u003eActa Physiologiae Plantarum\u003c/em\u003e 39 (8): 1\u0026ndash;11.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShalaby, Tarek, Yousry Bayoumi, Tarek Alshaal, Nevien Elhawat, Attila Sztrik, and Hassan El-Ramady. 2017. \u0026ldquo;Selenium Fortification Induces Growth, Antioxidant Activity, Yield and Nutritional Quality of Lettuce in Salt-Affected Soil Using Foliar and Soil Applications.\u0026rdquo; \u003cem\u003ePlant and Soil\u003c/em\u003e 421 (1\u0026ndash;2): 245\u0026ndash;58.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma, Pallavi, Ambuj Bhushan Jha, Rama Shanker Dubey, and Mohammad Pessarakli. 2012. \u0026ldquo;Reactive Oxygen Species, Oxidative Damage, and Antioxidative Defense Mechanism in Plants under Stressful Conditions.\u0026rdquo; \u003cem\u003eJournal of Botany\u003c/em\u003e 2012: 1\u0026ndash;26. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2012/217037\u003c/span\u003e\u003cspan address=\"10.1155/2012/217037\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShekari, Fariborz, Amin Abbasi, and Seyed Hamid Mustafavi. 2017. \u0026ldquo;Effect of Silicon and Selenium on Enzymatic Changes and Productivity of Dill in Saline Condition.\u0026rdquo; \u003cem\u003eJournal of the Saudi Society of Agricultural Sciences\u003c/em\u003e 16 (4): 367\u0026ndash;74.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSmirnoff, Nicholas. 1995. \u003cem\u003eEnvironment and Plant Metabolism: Flexibility and Acclimation\u003c/em\u003e. BIOS Scientific publishers.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSolecka, Danuta, and Alina Kacperska. 2003. \u0026ldquo;Phenylpropanoid Deficiency Affects the Course of Plant Acclimation to Cold.\u0026rdquo; \u003cem\u003ePhysiologia Plantarum\u003c/em\u003e 119 (2): 253\u0026ndash;62.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSpicher, Livia, Gaetan Glauser, and Felix Kessler. 2016. \u0026ldquo;Lipid Antioxidant and Galactolipid Remodeling under Temperature Stress in Tomato Plants.\u0026rdquo; \u003cem\u003eFrontiers in Plant Science\u003c/em\u003e 7: 167.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStepien, Piotr, and Grazyna Klobus. 2005. \u0026ldquo;Antioxidant Defense in the Leaves of C3 and C4 Plants under Salinity Stress.\u0026rdquo; \u003cem\u003ePhysiologia Plantarum\u003c/em\u003e 125 (1): 31\u0026ndash;40.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStudier, F William. 1973. \u0026ldquo;Analysis of Bacteriophage T7 Early RNAs and Proteins on Slab Gels.\u0026rdquo; \u003cem\u003eJournal of Molecular Biology\u003c/em\u003e 79 (2): 237\u0026ndash;48.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSullivan, Ch Y. 1979. \u0026ldquo;Selection for Drought and Heat Tolerance in Grain Sorghum.\u0026rdquo; \u003cem\u003eStress Physiology in Crop Plants\u003c/em\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSun, Z W, L K Ren, J W Fan, Q Li, K J Wang, M M Guo, L Wang, et al. 2016. \u0026ldquo;Salt Response of Photosynthetic Electron Transport System in Wheat Cultivars with Contrasting Tolerance.\u0026rdquo; \u003cem\u003ePlant, Soil and Environment\u003c/em\u003e 62 (11): 515\u0026ndash;21.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSuo, Jinwei, Qi Zhao, Lisa David, Sixue Chen, and Shaojun Dai. 2017. \u0026ldquo;Salinity Response in Chloroplasts: Insights from Gene Characterization.\u0026rdquo; \u003cem\u003eInternational Journal of Molecular Sciences\u003c/em\u003e 18 (5): 1011.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSzabados, Laszlo, and Arnould Savoure. 2010. \u0026ldquo;Proline: A Multifunctional Amino Acid.\u0026rdquo; \u003cem\u003eTrends in Plant Science\u003c/em\u003e 15 (2): 89\u0026ndash;97.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTanaka, Yoshiyuki, Makoto Matsuoka, Naoki Yamanoto, Yuko Ohashi, Yuriko Kano-Murakami, and Yoshihiro Ozeki. 1989. \u0026ldquo;Structure and Characterization of a CDNA Clone for Phenylalanine Ammonia-Lyase from Cut-Injured Roots of Sweet Potato.\u0026rdquo; \u003cem\u003ePlant Physiology\u003c/em\u003e 90 (4): 1403\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTavili, A, S Zare, A Enayati, and others. 2009. \u0026ldquo;Hydropriming, Ascorbic and Salicylic Acid Influence on Germination of Agropyron Elongatum Host. Seeds under Salt Stress.\u0026rdquo; \u003cem\u003eResearch Journal of Seed Science\u003c/em\u003e 2 (1): 16\u0026ndash;22.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTerzi, Rabiye, Neslihan Saruhan G\u0026uuml;ler, Funda G\u0026uuml;l G\u0026uuml;ven, and Asim Kadioglu. 2018. \u0026ldquo;Alpha Lipoic Acid Treatment Induces the Antioxidant System and Ameliorates Lipid Peroxidation in Maize Seedlings under Osmotic Stress.\u0026rdquo; \u003cem\u003eArchives of Biological Sciences\u003c/em\u003e 70 (3): 503\u0026ndash;11. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2298/ABS171218011T\u003c/span\u003e\u003cspan address=\"10.2298/ABS171218011T\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThipyapong, Piyada, and John C Steffens. 1997. \u0026ldquo;Tomato Polyphenol Oxidase (Differential Response of the Polyphenol Oxidase F Promoter to Injuries and Wound Signals).\u0026rdquo; \u003cem\u003ePlant Physiology\u003c/em\u003e 115 (2): 409\u0026ndash;18.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWahid, Abdul, and Alia Ghazanfar. 2006. \u0026ldquo;Possible Involvement of Some Secondary Metabolites in Salt Tolerance of Sugarcane.\u0026rdquo; \u003cem\u003eJournal of Plant Physiology\u003c/em\u003e 163 (7): 723\u0026ndash;30.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWakeel, Abdul, Muhammad Farooq, Manzoor Qadir, and Sven Schubert. 2011. \u0026ldquo;Potassium Substitution by Sodium in Plants.\u0026rdquo; \u003cem\u003eCritical Reviews in Plant Sciences\u003c/em\u003e 30 (4): 401\u0026ndash;13.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, Xiaoyuan, and Peter J Quinn. 2000. \u0026ldquo;The Location and Function of Vitamin E in Membranes.\u0026rdquo; \u003cem\u003eMolecular Membrane Biology\u003c/em\u003e 17 (3): 143\u0026ndash;56.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWu, Nan, Yuangang Zu, Yujie Fu, Yu Kong, Jintong Zhao, Xiaojuan Li, Ji Li, Michael Wink, and Thomas Efferth. 2010. \u0026ldquo;Antioxidant Activities and Xanthine Oxidase Inhibitory Effects of Extracts and Main Polyphenolic Compounds Obtained from Geranium Sibiricum L.\u0026rdquo; \u003cem\u003eJournal of Agricultural and Food Chemistry\u003c/em\u003e 58 (8): 4737\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYu, Xiao-Zhang, and Ji-Dong Gu. 2013. \u0026ldquo;Phyto-Transport and Assimilation of Selenium.\u0026rdquo; In \u003cem\u003ePlant-Based Remediation Processes\u003c/em\u003e, 159\u0026ndash;75. Springer.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZeid, I M, ZFAE Gharib, S M Ghazi, and E Z Ahmed. 2019. \u0026ldquo;Promotive Effect of Ascorbic Acid, Gallic Acid, Selenium and Nano-Selenium on Seed Germination, Seedling Growth and Some Hydrolytic Enzymes Activity of Cowpea (Vigna Unguiculata) Seedling.\u0026rdquo; \u003cem\u003eJ Plant Physiol Pathol 7\u003c/em\u003e 1: 1\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZong, Xiao-juan, Da-peng Li, Ling-kun Gu, De-quan Li, Li-xia Liu, and Xiao-li Hu. 2009. \u0026ldquo;Abscisic Acid and Hydrogen Peroxide Induce a Novel Maize Group C MAP Kinase Gene, ZmMPK7, Which Is Responsible for the Removal of Reactive Oxygen Species.\u0026rdquo; \u003cem\u003ePlanta\u003c/em\u003e 229 (3): 485\u0026ndash;95.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Selenium, α-tocopherol, Ascorbic acid, Salt stress, Protein, Antioxidants","lastPublishedDoi":"10.21203/rs.3.rs-8692316/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8692316/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSelenium, α-tocopherol, and ascorbic acid are important antioxidants that mitigate abiotic stresses in plants, yet their comparative effects under salinity remain insufficiently studied. This study aimed to evaluate and compare the effects of selenium, α-tocopherol, and ascorbic acid on growth, carbon metabolism, and ion uptake in salt-stressed maize plant. Deionised water (control), selenium (0.5 mM), α-tocopherol (200 ppm), or ascorbic acid (500 ppm) were applied to maize seeds, which were then cultivated at NaCl concentrations of 0, 100, 150, and 200 mM. Plant samples were examined for ionic, physiological and biochemical characteristics after 40 days. Salt stress caused concentration-dependent reductions in growth, chlorophyll, insoluble sugars, carbohydrates, phenolics, flavonoids, and ion uptake, while enhancing soluble sugars, α-amylase activity, and sodium accumulation. On the other hand, growth performance, pigment content, carbohydrate and secondary metabolites, and the K⁺/Na⁺ and Ca\u0026sup2;⁺/Na⁺ ratios were all improved by antioxidant treatments administered in non-stressful environments. Among the tested compounds, ascorbic acid was most effective in alleviating salt-induced damage. It enhanced antioxidant activity, improved carbon metabolism and ion homeostasis, and induced proteins linked to salt tolerance, leading to better growth under salinity. In conclusion, ascorbic acid shows promise as a way to reduce salt stress and increase maize yields in salty soils.\u003c/p\u003e","manuscriptTitle":"Differential Modulation of Antioxidant Defense and Salinity Tolerance in Maize by Selenium, α-Tocopherol, and Ascorbic Acid","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-23 17:21:42","doi":"10.21203/rs.3.rs-8692316/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2026-05-13T10:10:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"59313591101685323507147183873472712743","date":"2026-05-04T13:07:00+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-30T11:35:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"108094429769188193466699739151912809581","date":"2026-04-20T23:11:21+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-08T07:12:36+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"295866983126246458534801756853020944224","date":"2026-03-18T09:03:20+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"160577388117156105332515991141941257639","date":"2026-03-01T18:07:26+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-19T07:42:49+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-04T10:47:52+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-02-04T10:35:47+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-02-03T12:48:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2026-02-03T12:03:35+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"fa3ae6d5-32ef-400e-8b95-e3b7df098562","owner":[],"postedDate":"February 23rd, 2026","published":true,"recentEditorialEvents":[{"type":"editorInvitedReview","content":"","date":"2026-05-13T10:10:48+00:00","index":78,"fulltext":""},{"type":"reviewerAgreed","content":"59313591101685323507147183873472712743","date":"2026-05-04T13:07:00+00:00","index":75,"fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-30T11:35:24+00:00","index":69,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":63182548,"name":"Biological sciences/Biochemistry"},{"id":63182549,"name":"Biological sciences/Physiology"},{"id":63182550,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2026-02-23T17:21:43+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-23 17:21:42","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8692316","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8692316","identity":"rs-8692316","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}