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Multivariate Analysis to Evaluate the Effectiveness of using Diluted Seawater to Increase Wheat’s Resistance to Ionic Stress

H
Hamida Sadji-Ait Kaci1,*
A
Assia Chaker-Haddadj1
F
Farida Taguett1
F
Fahima Nabi2
M
Massakib Bekkaye1
R
Rafika Ghanai3
N
Nesrine Bedjeloud1
A
Amina Bekkouche2
1Laboratory of Biology and Organism of Physiology, Faculty of Biological Sciences, University of Sciences and Technology Houari Boumediene, Team of Soils Biology, Algiers, Algeria.
2Laboratory of Biotechnology and Valorization of Biological Resources, Faculty of Sciences, Yahia Fares Medea University, Ouzra, Medea 26100, Algeria.
3Université Saad Dahleb, Blida 1, Algeria.

ABSTRACT

Background: Freshwater scarcity is increasingly threatening sustainable agricultural production, highlighting the need for alternative irrigation resources. This study aimed to evaluate the effects of irrigation with phosphate-enriched diluted seawater on wheat resilience to stressful conditions.

Methods: Nine diluted seawaters (DSW) with or without two levels of phosphate (50T and 90T): Control (freshwater), W10 (10% of seawater), W20 (20% of seawater), W40 (40% of seawater), W10T50, W10T90, W20T50, W20T90, W10T90, W40T50, W40T90 were used to study the germination, growth and physiological responses of wheat plants. Physical and chemical quality of the DSW were also estimated to assess their potential agronomic impact.

Result: The results indicated that the treated DSW used presented a notable permeability index, kelly’s index with low hardness. No significant differences were observed in germination traits under W10, W20 and W40 treatments compared to control. The aerial biomass was significantly (p<0.05) reduced, while root part was significantly increased under all treatments. Application of phosphate to W10 and W20 increased the chlorophyll pigments (+40%), the relative water content exceeded 93% and decreased electrolytes leakage (76%). Heatmap and PCA analysis identified three clusters: (A) related to germination traits under C, W10 and W10T treatments and carbonate ions, (B) physiological traits under W20 and W20T and (C) osmotic adjustment markers phosphorus ions. Therefore, diluted seawater enriched with phosphate may serve as a viable solution for water scarcity and soil health in arid regions.

INTRODUCTION

Food security and biodiversity protection are two major issues requiring an in-depth scientific investigation to improve agricultural and environmental interventions. The world population will exceed 9 billion by 2050 and will need more than a 70% increase in food production, in order to meet growing global food demand (Sarfraz et al., 2023). Whereas, unavailability of freshwater is becoming real threat for agriculture due to growing global populations, climate change and depleting traditional water supplies (FAO, 2023). According to Ashik et al. (2023), to ensure this demand, agricultural production must increase by 50% by 2025. Triticum aestivum L. is an essential cereal species, providing a significant supply of carbs, proteins, dietary fibers, vitamins and bioactive substances. It is mostly grown in winter season which led wheat plants to suffer with salinity stress (Ashik et al., 2023). Salinity is a prominent abiotic stress that limits the growth and productivity of crops influencing physiological characteristics and oxidative stress through the generation of reactive oxygen species (ROS), resulting in cellular damage and photosynthetic efficiency (Ahanger et al., 2023).
       
The use of diluted seawater could be sustainable alternative allowing tolerance capacity of stressed plants. However, the high NaCl content and phosphate scarcity in seawater involving a preliminary investigation for irrigation, ensuring plant resistance to salt stress and avoiding soil sodicity. Researches are focused on use of diluted seawater in agriculture but few or no studies have examined the potential effects of diluted seawater (DSW) enriched with phosphate plant resilience to salt stress. In fact, our prior research has demonstrated that the application of phosphate to chickpea plants has improved crop yield and salinity tolerance by over 30% (Sadji-Ait et al., 2018).  Thus, we hypothesize that diluted seawater (DSW) could serve as a sustainable agricultural water source and the main objective of this study is to investigate the  advantages of adding phosphate fertilizer to diluted seawater on germination performance and morpho-physiological attributes of wheat plants. The findings could contribute to sustainable agronomic practices by leveraging seawater availability.

MATERIALS AND METHODS

The study was carried out during 2024-2025, at the laboratory of Soil’s Biology-University of Sciences and Technology Houari Boumediene, Algeria. The seawater used in this study was took from Collo located in Northeastern Algeria (37°00′23″North, 6°33′39″Est).
 
Physical and chemical determination
 
The chemical and physical proprieties of the used seawater were investigated and presented in Table 1. The pH and EC were measured by using a portable pH meter. Ten treatments used were prepared in bottles using by addition of distilled water to the collected seawater in order to assess germination and wheat plants growth. Treatments were divided in two blocks: first block composed by diluted seawater (DSW) by addition of distilled water the seawater: C (distilled water: control), W10 (10% of seawater + 90% of distilled water), W20 (20% of seawater completed with 80% of distilled water) and W40 (40% of seawater completed with 60% of distilled water). Second block was composed by the previous diluted seawater with two concentrations of phosphate separately (T): 0.05% and 0.09% corresponding to 0.05 mg.L-1 and 0.09 mg.L-1 of phosphate respectively. These concentrations were studied to evaluated their efficacity in agriculture. Thus, total treatments used were:  C, W10, W20, W40, W10T50, W10T90, W20T50, W20T90, W40T50 and W40T90. Finally, samples were taken from each treatment in order to assess some physical and chemical traits (Table 1).

Table 1: Chemical and physical proprieties of fresh water, seawater, and diluted seawater (DSW) with or without phosphate addition (0.5% and 0.9%) compared to initial seawater characteristics (Rodier, 2009).


 
Ratio of sodium adsorption (SAR)
 
The sodicity of the soil is determined by the sodium adsorption ratio. The following formula was used to determine the SAR (USDA, 1954).
       
Sodium adsorption ratio (SAR) was measured using the following equation (USDA, 1954):
      
SAR = Na+ + √(Ca2+ + Mg2+)/2
 
Permeability index (PI)
 
Permeability index (PI) indicated the effect if the water is suitability for irrigation usage (Doneen, 1964) using the following formula:       
 
PI = Na+ + √HCO3- (Na+ + K+ + Mg2+ + Ca2+) *100
 
The PI classification is as follows: Excellent (>75%), Good (25-75%) and unsuitable (<25%) (Al-Amry, 2008).
 
Kelly’s ratio (KR)
 
Kelly’s ratio (KR) is quality index reveal the sodicity hazard in soils and evaluated using following formula (Kelly, 1940):
 
KI = Na+ + (Ca2+ + Mg2+)
 
Water with a KI≤1 is recommended for irrigation, while water with KI≥1 is not recommended for irrigation.
 
Water hardness
 
The hardness of seawater treatments (TH) was calculated using following formula and classification (Todd, 1980):  
 
TH (°F) = 2.5 × Ca2+ + 4.1 × Mg2+ 
 
Soft water (TH<75), moderately hard (TH 75-150), hard water (TH 150-300) and very hard water (TH>300).
 
Germination test design
 
Wheat seeds (cv Maouna) were disinfected using 1% sodium hypochlorite for three minutes, then rinsed with distilled water. Seeds were placed in Petri dishes between two layers of filter paper and soaked in the treatment solution. Three replicates (Petri dishes) were used for each treatment (Bekele et al., 2024). Twenty were placed in seeds in each Petri dish resulting thirty experimental unit and six hundred seeds were used. The seeds were kept at 25±0.5°C in darkness for 7 days according to randomised complete block design (CRBD).
 
Germination percentage (GP)
 
After 7 days, germination rate corresponds to percentage of the number of sprouted seeds divided on total number of seeds, until the number remained constant (Maguire, 1962).


 
Germination speed
 
Speed germination was estimated according to Rubio-Casal et al. (2003) formula:   
 
SG = N1/D1 + N2/D2 + N3/D3 +……+ Nn/Dn
 
Where,
N1= The number of seeds germinated in day one (D1) of sowing.
 
Seed vigour index
 
Five randomly selected seedlings from each treatment were divided into root and shoot components, lengths and fresh weights were measured and dried at 70°C for 72 hours to obtain dry weights for both parts. The vigour index (SVI) was calculated using AOSA, (1983) method.
 

 
Plant growth setup
 
After complete emergence, five seedlings were taken from each treatment and transferred into pots containing 700 g of soil. Physical and chemical characteristics of the soil are: clay (44%), silt (41%), sandy (7.5%) with pH (7) and EC (0.18 mhoms/cm). Chemical composition was: phosphate (34.5 ppm), potassium (270 ppm), calcium (0.13 meq.G-1), sodium (0.31 meq.G-1), nitrogen (0.12%), organic matter (1.75%), organic carbon (0.98%).
       
The seedlings were irrigated once week using DSW added or no with phosphate as described before C, W10, W20, W40, W10T50, W10T90, W20T50, W20T90, W40T50 and W40T90. Pots were placed at ambient temperature with a day’s photoperiod of 16h/8h during sixty days.
 
Physiological parameters
 
At 60th day after sowing, samples of plants were taken from each treatment to assess growth traits: stem height, root length, stem dry weights and root dry weight.
       
Relative water content (RWC) was determined based on the fresh weight (FW), turgid weight (TW) and dry weight (DW) of plants according to Ates and Tekeli (2007):
 

 
Photosynthetic pigments
 
Pigment were extracted from fresh tissue with 80% acetone. The absorbance was measured at 663, 645 and 480 nm using a spectrophotometer according (Lichtenthaler, 1987):
 
Chlorophyll a (µg/mL) = (11.75 × A663 - 2.35 × A645)
 
 Chlorophyll b (µg/mL) = (18.61 × A645 - 3.96 × A663)
 
 Carotenoid (µg/mL) = (1000 × A470) - (2.27 × Chla) - (81.4 × Chlb)/227)
 
Proline determination
 
Twenty μL of 1% sulfosalicylic acid was applied to 0.5 g of fresh leaves. The supernatant was collected and mixed in a 1:2 ratio with acidic ninhydrin (1.25% ninhydrin in 80% acetic acid) and centrifugated at 15,000 g for 5 minutes. The mixture was incubated for 30 minutes at 95°C to measure the absorbance at 510 nm using spectrophotometer (Bates et al., 1973).
 
Electrolytes leakage
 
Stability of cellular membranes was estimated basing on the electrolyte leakage using the following formula (Murray et al., 1989):
 
EL (%) = (ECi/ECf) × 100%
 
Where,
ECi= t initial electrical conductivity and the (ECf) to final electrical conductivity.
 
Statistical analysis
 
Five replicates of each treatment were used to study plant parameters according to experimental complete randomized block design. The means and standard errors (shown in tables and graphs) were calculated using Excel version 2019. Data were analysed by ANOVA followed by the LSD post hoc test (p≤0.05).
       
Principal component analyses (PCA) and Heatmap clustering were made by thirty normalized variables, including the germination performances, morphological, physiological and hydraulic parameters of plants, physical and chemical characteristics of treated diluted seawater from each treatment to designate the main factors impacting the seeds and plant responses to salt stress and relationship between them and nutritional component of diluted seawater using statistica software package.

RESULTS AND DISCUSSION

Chemical examination of the used diluted seawater revealed a good permeability index (0.95 to 1.05) and Kelly’s index, indicating that the water hardness level was soft water, notably W10T and W20T, as characterized by Todd (1980). Maintaining water levels and proper soil penetration under the W10, W20 and W40 treatments may improve plant nutrition by assuring appropriate mineral distribution. Key cations such as potassium, magnesium and calcium help to achieve nutritional balance and counterbalance the negative effects of excessive sodium by controlling ionic homeostasis (Zrig et al., 2021). The concentrations of bicarbonates (HCO3-) do not change significantly across treatments using (DSW±T). This stability could be positive for the colligative propriety of ions, electrical conductivity and pH of irrigation solution are critical factors in crop cultivation because they are instable due to nutrient uptake (Ekman et al., 1945) (Table 1). Amer et al. (2017) have showed that the chemical characteristics of the irrigated soil with diluted seawater with compost were remarkably affected the salinity accumulation around the root zone in the second season, but lesser extent than during the first season. Furthermore, they reported that the diluted seawater applications had lowest effect on the available macronutrient contents of the soil compared to the effect of the compost addition as individual factors.
       
None significant differences were observed in germination rate of wheat seeds under all treatments, while the (DSW+T) treatment significantly (p≤0.05) decreased the speed germination, the seedlings traits and vigor index by about 15%, 38% and 13%, respectively compared to fresh water used as control (Table 2). It is probably related to caused by the suppression of enzyme processes involved in starch mobilization and embryo development mobilization and reduced mitochondrial activity during early growth (Chen et al., 2022; Nabi et al., 2025, Bouchenafa et al., 2026). Indira et al., (2026) have found that decline of chickpea germination could be related to increase of the proteases activity under osmotic stress compared to control. Results indicated that DSW, DSW+T50 and DSW+T90 significantly improved root dry weight by about 25%, 88% and 68% respectively compared to water.  Remarkably, a high and significant (p≤0.05) improvement of plant growth parameters was noted under W40T90 treatment inducing maximum stem length (28%) and dry weight (38%) compared to plants control (Table 2).

Table 2: Effects of different of diluted seawater (DSW) with or without phosphate (T) on germination percentage (GP), speed germination (SG), seedlings growth, (Shoot length (LT), root length (LR) vigor index (IV), plant growth and photosynthetic pigments contents of Triticum aestivum L.cv. Maouna plants.


       
Physiologically, the relative water content (RWC) was significantly decreased by an average of  (60.07%)) in W20 plants, while it improved by about 93% in W10 and W40 compared to plant control (Fig 1). Plants control revealed 74.8% of electrolyte leakage (EL) which was significantly (p≤0.05) higher than those indicated by plants irrigated other treatments. The EL was less than 35%, 25% and 15% for DSW, DSW+T and DSW+T plants respectively. Comparison to fresh water, an irrigation of plants with DSW exhibited an increase of Chla content from 0.18 µg.G-1 to more than 0.29 µg.G particularly under W10 and W20 treatments. The W40 treatment induced the lowest level of total Chlorophyll. These results could be due to high concentration of NaCl (Chaker-Haddadj et al., 2014; Fauzan et al. (2026). They stipulated that prolonged exposure to NaCl and high levels in tissue culture media can result in necrosis (cell death) of apical buds and young leaf tissue.

Fig 1: Effects of different of diluted seawater (DSW) added or no with or without.


       
Whereas, the W20T50 and DSW+T90 treatments increased the Chla to achieve a maximum of 0.86 µg.G-1. The Chlb content was significantly increased from 0.79 µg.G-1 to more than 0.97 µg.G-1 particularly for W10, W10T50 and W10T90 (Table 2). Carotenoid content decreased with increase of salt stress and low P application (WT50), but it significantly increased when high phosphorus dose was added to diluted seawater WT90, suggesting a negative effects of these treatments on plants behavior. An increase of carotenoids would have antioxidants effects could,, prevent photo-oxidative damage by scavenging ROS generated under abiotic stress (Alinia et al., 2024). Sadji-Ait et al. (2022) showed that phosphorus application has mitigated salt stress by reducing oxidative stress through increase of H2O2 activity in chickpea genotypes under salinity. Moreover, endogenous proline content was lowest in (W10±T) plants but increased more than 78% in (W40±T90) plants (Fig 1). These results corroborated those found in some wheat genotypes by Yadav et al. (2021) would due to a strategy of controlling metabolic activities and energy efficiency during germination to fulfill subsequent needs during plant growth significantly increased than the plant control.
       
The principal component analysis (PCA) identified a positive correlation between stress tolerance indicators: proline content, carotenoids and relative dry weight with mineral contents in seawater treatments (W20, W20T, W40, W40T) on the F1 axis. Conversely, germination parameters, photosynthetic pigments, cell turgor and membrane integrity exhibited negative correlations with treatments (C, W10, W10T). The second principal component (F2) indicated an independent and antagonistic relationship among these traits.  Heatmap analyses revealed three clusters: (A) germination traits related to hydrogen carbonate ion (HCO3) under control and low stress, (B) physiological plant traits with orthophosphate ions (HPO-24) linked to moderate stress and (C) high Na+ and Cl- content and osmotic adjustment markers under severe stress (Fig 2).

Fig 2: Biplot of principal components and heatmap analysis of thirty traits related to the treatments applied on soil characters, growth and physiological parameters of Triticum aestivumL. cv. maouna plants.


       
The multivariate analysis (p≤0.05) revealed that treatments with low carbonate levels in solutions W10, W10T50 and W10T90 positively impacted germination traits, akin to fresh water control (Fig 2). This result could be  attributed tos a hormetic effect where mild alkaline stress enhanced metabolic processes vital for germination as reported by Agathokleous et al., (2024). Additionally, seeds’ ability to maintain cellular homeostasis during imbibition and the seed coat’s buffering action likely mitigate the effects of external pH on the embryo, contributing to this tolerance. The W20T  treatment induced an improvement of cell hydraulic status and photosynthetic pigment levels while reducing membrane disruption and proline accumulation.
       
These responses could suggest a physiological stability and  an efficiency of the photosynthetic enzyme system such as Rubisco activity and protein content relative to maintain optimal physiological functioning under stress (Wang et al., 2026). 

CONCLUSION

This study has demonstrated that diluted seawater (DSW) can serve as a viable irrigation source in agriculture, exhibiting minimal negative effects on plant development. Irrigation of wheat with DSW (at 10%, 20% and 40%) resulted in low concentrations of minerals beneficial for germination, positively correlated with carbonate ions. Additionally, incorporating phosphate into DSW enhanced salinity tolerance and was correlated with better plant growth and tolerance particularly with 0.9% of phosphate. Thus, the use of diluted seawater enriched with optimized doses of phosphate could represent a promising approach for improving crop yields.  

ACKNOWLEDGEMENT

The present study was supported by no external funding.
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
 
Author contributions
 
Author Hamida Sadji-Ait Kaci: Conceptualization, methodology, investigation, writing original draft. Author Assia Chaker-Haddadj, Farida Taguett and Rafika Ghanai:  Validation, visualization, Author Massakib Bekkaye and Nabi: statistical analysis and data curation, Massakib Bekkaye and Amina Bekkouche: Supervision and visualization. All authors have agreed to the published version of the manuscript.

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

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    Full Research Article

    Multivariate Analysis to Evaluate the Effectiveness of using Diluted Seawater to Increase Wheat’s Resistance to Ionic Stress

    H
    Hamida Sadji-Ait Kaci1,*
    A
    Assia Chaker-Haddadj1
    F
    Farida Taguett1
    F
    Fahima Nabi2
    M
    Massakib Bekkaye1
    R
    Rafika Ghanai3
    N
    Nesrine Bedjeloud1
    A
    Amina Bekkouche2
    1Laboratory of Biology and Organism of Physiology, Faculty of Biological Sciences, University of Sciences and Technology Houari Boumediene, Team of Soils Biology, Algiers, Algeria.
    2Laboratory of Biotechnology and Valorization of Biological Resources, Faculty of Sciences, Yahia Fares Medea University, Ouzra, Medea 26100, Algeria.
    3Université Saad Dahleb, Blida 1, Algeria.

    ABSTRACT

    Background: Freshwater scarcity is increasingly threatening sustainable agricultural production, highlighting the need for alternative irrigation resources. This study aimed to evaluate the effects of irrigation with phosphate-enriched diluted seawater on wheat resilience to stressful conditions.

    Methods: Nine diluted seawaters (DSW) with or without two levels of phosphate (50T and 90T): Control (freshwater), W10 (10% of seawater), W20 (20% of seawater), W40 (40% of seawater), W10T50, W10T90, W20T50, W20T90, W10T90, W40T50, W40T90 were used to study the germination, growth and physiological responses of wheat plants. Physical and chemical quality of the DSW were also estimated to assess their potential agronomic impact.

    Result: The results indicated that the treated DSW used presented a notable permeability index, kelly’s index with low hardness. No significant differences were observed in germination traits under W10, W20 and W40 treatments compared to control. The aerial biomass was significantly (p<0.05) reduced, while root part was significantly increased under all treatments. Application of phosphate to W10 and W20 increased the chlorophyll pigments (+40%), the relative water content exceeded 93% and decreased electrolytes leakage (76%). Heatmap and PCA analysis identified three clusters: (A) related to germination traits under C, W10 and W10T treatments and carbonate ions, (B) physiological traits under W20 and W20T and (C) osmotic adjustment markers phosphorus ions. Therefore, diluted seawater enriched with phosphate may serve as a viable solution for water scarcity and soil health in arid regions.

    INTRODUCTION

    Food security and biodiversity protection are two major issues requiring an in-depth scientific investigation to improve agricultural and environmental interventions. The world population will exceed 9 billion by 2050 and will need more than a 70% increase in food production, in order to meet growing global food demand (Sarfraz et al., 2023). Whereas, unavailability of freshwater is becoming real threat for agriculture due to growing global populations, climate change and depleting traditional water supplies (FAO, 2023). According to Ashik et al. (2023), to ensure this demand, agricultural production must increase by 50% by 2025. Triticum aestivum L. is an essential cereal species, providing a significant supply of carbs, proteins, dietary fibers, vitamins and bioactive substances. It is mostly grown in winter season which led wheat plants to suffer with salinity stress (Ashik et al., 2023). Salinity is a prominent abiotic stress that limits the growth and productivity of crops influencing physiological characteristics and oxidative stress through the generation of reactive oxygen species (ROS), resulting in cellular damage and photosynthetic efficiency (Ahanger et al., 2023).
           
    The use of diluted seawater could be sustainable alternative allowing tolerance capacity of stressed plants. However, the high NaCl content and phosphate scarcity in seawater involving a preliminary investigation for irrigation, ensuring plant resistance to salt stress and avoiding soil sodicity. Researches are focused on use of diluted seawater in agriculture but few or no studies have examined the potential effects of diluted seawater (DSW) enriched with phosphate plant resilience to salt stress. In fact, our prior research has demonstrated that the application of phosphate to chickpea plants has improved crop yield and salinity tolerance by over 30% (Sadji-Ait et al., 2018).  Thus, we hypothesize that diluted seawater (DSW) could serve as a sustainable agricultural water source and the main objective of this study is to investigate the  advantages of adding phosphate fertilizer to diluted seawater on germination performance and morpho-physiological attributes of wheat plants. The findings could contribute to sustainable agronomic practices by leveraging seawater availability.

    MATERIALS AND METHODS

    The study was carried out during 2024-2025, at the laboratory of Soil’s Biology-University of Sciences and Technology Houari Boumediene, Algeria. The seawater used in this study was took from Collo located in Northeastern Algeria (37°00′23″North, 6°33′39″Est).
     
    Physical and chemical determination
     
    The chemical and physical proprieties of the used seawater were investigated and presented in Table 1. The pH and EC were measured by using a portable pH meter. Ten treatments used were prepared in bottles using by addition of distilled water to the collected seawater in order to assess germination and wheat plants growth. Treatments were divided in two blocks: first block composed by diluted seawater (DSW) by addition of distilled water the seawater: C (distilled water: control), W10 (10% of seawater + 90% of distilled water), W20 (20% of seawater completed with 80% of distilled water) and W40 (40% of seawater completed with 60% of distilled water). Second block was composed by the previous diluted seawater with two concentrations of phosphate separately (T): 0.05% and 0.09% corresponding to 0.05 mg.L-1 and 0.09 mg.L-1 of phosphate respectively. These concentrations were studied to evaluated their efficacity in agriculture. Thus, total treatments used were:  C, W10, W20, W40, W10T50, W10T90, W20T50, W20T90, W40T50 and W40T90. Finally, samples were taken from each treatment in order to assess some physical and chemical traits (Table 1).

    Table 1: Chemical and physical proprieties of fresh water, seawater, and diluted seawater (DSW) with or without phosphate addition (0.5% and 0.9%) compared to initial seawater characteristics (Rodier, 2009).


     
    Ratio of sodium adsorption (SAR)
     
    The sodicity of the soil is determined by the sodium adsorption ratio. The following formula was used to determine the SAR (USDA, 1954).
           
    Sodium adsorption ratio (SAR) was measured using the following equation (USDA, 1954):
          
    SAR = Na+ + √(Ca2+ + Mg2+)/2
     
    Permeability index (PI)
     
    Permeability index (PI) indicated the effect if the water is suitability for irrigation usage (Doneen, 1964) using the following formula:       
     
    PI = Na+ + √HCO3- (Na+ + K+ + Mg2+ + Ca2+) *100
     
    The PI classification is as follows: Excellent (>75%), Good (25-75%) and unsuitable (<25%) (Al-Amry, 2008).
     
    Kelly’s ratio (KR)
     
    Kelly’s ratio (KR) is quality index reveal the sodicity hazard in soils and evaluated using following formula (Kelly, 1940):
     
    KI = Na+ + (Ca2+ + Mg2+)
     
    Water with a KI≤1 is recommended for irrigation, while water with KI≥1 is not recommended for irrigation.
     
    Water hardness
     
    The hardness of seawater treatments (TH) was calculated using following formula and classification (Todd, 1980):  
     
    TH (°F) = 2.5 × Ca2+ + 4.1 × Mg2+ 
     
    Soft water (TH<75), moderately hard (TH 75-150), hard water (TH 150-300) and very hard water (TH>300).
     
    Germination test design
     
    Wheat seeds (cv Maouna) were disinfected using 1% sodium hypochlorite for three minutes, then rinsed with distilled water. Seeds were placed in Petri dishes between two layers of filter paper and soaked in the treatment solution. Three replicates (Petri dishes) were used for each treatment (Bekele et al., 2024). Twenty were placed in seeds in each Petri dish resulting thirty experimental unit and six hundred seeds were used. The seeds were kept at 25±0.5°C in darkness for 7 days according to randomised complete block design (CRBD).
     
    Germination percentage (GP)
     
    After 7 days, germination rate corresponds to percentage of the number of sprouted seeds divided on total number of seeds, until the number remained constant (Maguire, 1962).


     
    Germination speed
     
    Speed germination was estimated according to Rubio-Casal et al. (2003) formula:   
     
    SG = N1/D1 + N2/D2 + N3/D3 +……+ Nn/Dn
     
    Where,
    N1= The number of seeds germinated in day one (D1) of sowing.
     
    Seed vigour index
     
    Five randomly selected seedlings from each treatment were divided into root and shoot components, lengths and fresh weights were measured and dried at 70°C for 72 hours to obtain dry weights for both parts. The vigour index (SVI) was calculated using AOSA, (1983) method.
     

     
    Plant growth setup
     
    After complete emergence, five seedlings were taken from each treatment and transferred into pots containing 700 g of soil. Physical and chemical characteristics of the soil are: clay (44%), silt (41%), sandy (7.5%) with pH (7) and EC (0.18 mhoms/cm). Chemical composition was: phosphate (34.5 ppm), potassium (270 ppm), calcium (0.13 meq.G-1), sodium (0.31 meq.G-1), nitrogen (0.12%), organic matter (1.75%), organic carbon (0.98%).
           
    The seedlings were irrigated once week using DSW added or no with phosphate as described before C, W10, W20, W40, W10T50, W10T90, W20T50, W20T90, W40T50 and W40T90. Pots were placed at ambient temperature with a day’s photoperiod of 16h/8h during sixty days.
     
    Physiological parameters
     
    At 60th day after sowing, samples of plants were taken from each treatment to assess growth traits: stem height, root length, stem dry weights and root dry weight.
           
    Relative water content (RWC) was determined based on the fresh weight (FW), turgid weight (TW) and dry weight (DW) of plants according to Ates and Tekeli (2007):
     

     
    Photosynthetic pigments
     
    Pigment were extracted from fresh tissue with 80% acetone. The absorbance was measured at 663, 645 and 480 nm using a spectrophotometer according (Lichtenthaler, 1987):
     
    Chlorophyll a (µg/mL) = (11.75 × A663 - 2.35 × A645)
     
     Chlorophyll b (µg/mL) = (18.61 × A645 - 3.96 × A663)
     
     Carotenoid (µg/mL) = (1000 × A470) - (2.27 × Chla) - (81.4 × Chlb)/227)
     
    Proline determination
     
    Twenty μL of 1% sulfosalicylic acid was applied to 0.5 g of fresh leaves. The supernatant was collected and mixed in a 1:2 ratio with acidic ninhydrin (1.25% ninhydrin in 80% acetic acid) and centrifugated at 15,000 g for 5 minutes. The mixture was incubated for 30 minutes at 95°C to measure the absorbance at 510 nm using spectrophotometer (Bates et al., 1973).
     
    Electrolytes leakage
     
    Stability of cellular membranes was estimated basing on the electrolyte leakage using the following formula (Murray et al., 1989):
     
    EL (%) = (ECi/ECf) × 100%
     
    Where,
    ECi= t initial electrical conductivity and the (ECf) to final electrical conductivity.
     
    Statistical analysis
     
    Five replicates of each treatment were used to study plant parameters according to experimental complete randomized block design. The means and standard errors (shown in tables and graphs) were calculated using Excel version 2019. Data were analysed by ANOVA followed by the LSD post hoc test (p≤0.05).
           
    Principal component analyses (PCA) and Heatmap clustering were made by thirty normalized variables, including the germination performances, morphological, physiological and hydraulic parameters of plants, physical and chemical characteristics of treated diluted seawater from each treatment to designate the main factors impacting the seeds and plant responses to salt stress and relationship between them and nutritional component of diluted seawater using statistica software package.

    RESULTS AND DISCUSSION

    Chemical examination of the used diluted seawater revealed a good permeability index (0.95 to 1.05) and Kelly’s index, indicating that the water hardness level was soft water, notably W10T and W20T, as characterized by Todd (1980). Maintaining water levels and proper soil penetration under the W10, W20 and W40 treatments may improve plant nutrition by assuring appropriate mineral distribution. Key cations such as potassium, magnesium and calcium help to achieve nutritional balance and counterbalance the negative effects of excessive sodium by controlling ionic homeostasis (Zrig et al., 2021). The concentrations of bicarbonates (HCO3-) do not change significantly across treatments using (DSW±T). This stability could be positive for the colligative propriety of ions, electrical conductivity and pH of irrigation solution are critical factors in crop cultivation because they are instable due to nutrient uptake (Ekman et al., 1945) (Table 1). Amer et al. (2017) have showed that the chemical characteristics of the irrigated soil with diluted seawater with compost were remarkably affected the salinity accumulation around the root zone in the second season, but lesser extent than during the first season. Furthermore, they reported that the diluted seawater applications had lowest effect on the available macronutrient contents of the soil compared to the effect of the compost addition as individual factors.
           
    None significant differences were observed in germination rate of wheat seeds under all treatments, while the (DSW+T) treatment significantly (p≤0.05) decreased the speed germination, the seedlings traits and vigor index by about 15%, 38% and 13%, respectively compared to fresh water used as control (Table 2). It is probably related to caused by the suppression of enzyme processes involved in starch mobilization and embryo development mobilization and reduced mitochondrial activity during early growth (Chen et al., 2022; Nabi et al., 2025, Bouchenafa et al., 2026). Indira et al., (2026) have found that decline of chickpea germination could be related to increase of the proteases activity under osmotic stress compared to control. Results indicated that DSW, DSW+T50 and DSW+T90 significantly improved root dry weight by about 25%, 88% and 68% respectively compared to water.  Remarkably, a high and significant (p≤0.05) improvement of plant growth parameters was noted under W40T90 treatment inducing maximum stem length (28%) and dry weight (38%) compared to plants control (Table 2).

    Table 2: Effects of different of diluted seawater (DSW) with or without phosphate (T) on germination percentage (GP), speed germination (SG), seedlings growth, (Shoot length (LT), root length (LR) vigor index (IV), plant growth and photosynthetic pigments contents of Triticum aestivum L.cv. Maouna plants.


           
    Physiologically, the relative water content (RWC) was significantly decreased by an average of  (60.07%)) in W20 plants, while it improved by about 93% in W10 and W40 compared to plant control (Fig 1). Plants control revealed 74.8% of electrolyte leakage (EL) which was significantly (p≤0.05) higher than those indicated by plants irrigated other treatments. The EL was less than 35%, 25% and 15% for DSW, DSW+T and DSW+T plants respectively. Comparison to fresh water, an irrigation of plants with DSW exhibited an increase of Chla content from 0.18 µg.G-1 to more than 0.29 µg.G particularly under W10 and W20 treatments. The W40 treatment induced the lowest level of total Chlorophyll. These results could be due to high concentration of NaCl (Chaker-Haddadj et al., 2014; Fauzan et al. (2026). They stipulated that prolonged exposure to NaCl and high levels in tissue culture media can result in necrosis (cell death) of apical buds and young leaf tissue.

    Fig 1: Effects of different of diluted seawater (DSW) added or no with or without.


           
    Whereas, the W20T50 and DSW+T90 treatments increased the Chla to achieve a maximum of 0.86 µg.G-1. The Chlb content was significantly increased from 0.79 µg.G-1 to more than 0.97 µg.G-1 particularly for W10, W10T50 and W10T90 (Table 2). Carotenoid content decreased with increase of salt stress and low P application (WT50), but it significantly increased when high phosphorus dose was added to diluted seawater WT90, suggesting a negative effects of these treatments on plants behavior. An increase of carotenoids would have antioxidants effects could,, prevent photo-oxidative damage by scavenging ROS generated under abiotic stress (Alinia et al., 2024). Sadji-Ait et al. (2022) showed that phosphorus application has mitigated salt stress by reducing oxidative stress through increase of H2O2 activity in chickpea genotypes under salinity. Moreover, endogenous proline content was lowest in (W10±T) plants but increased more than 78% in (W40±T90) plants (Fig 1). These results corroborated those found in some wheat genotypes by Yadav et al. (2021) would due to a strategy of controlling metabolic activities and energy efficiency during germination to fulfill subsequent needs during plant growth significantly increased than the plant control.
           
    The principal component analysis (PCA) identified a positive correlation between stress tolerance indicators: proline content, carotenoids and relative dry weight with mineral contents in seawater treatments (W20, W20T, W40, W40T) on the F1 axis. Conversely, germination parameters, photosynthetic pigments, cell turgor and membrane integrity exhibited negative correlations with treatments (C, W10, W10T). The second principal component (F2) indicated an independent and antagonistic relationship among these traits.  Heatmap analyses revealed three clusters: (A) germination traits related to hydrogen carbonate ion (HCO3) under control and low stress, (B) physiological plant traits with orthophosphate ions (HPO-24) linked to moderate stress and (C) high Na+ and Cl- content and osmotic adjustment markers under severe stress (Fig 2).

    Fig 2: Biplot of principal components and heatmap analysis of thirty traits related to the treatments applied on soil characters, growth and physiological parameters of Triticum aestivumL. cv. maouna plants.


           
    The multivariate analysis (p≤0.05) revealed that treatments with low carbonate levels in solutions W10, W10T50 and W10T90 positively impacted germination traits, akin to fresh water control (Fig 2). This result could be  attributed tos a hormetic effect where mild alkaline stress enhanced metabolic processes vital for germination as reported by Agathokleous et al., (2024). Additionally, seeds’ ability to maintain cellular homeostasis during imbibition and the seed coat’s buffering action likely mitigate the effects of external pH on the embryo, contributing to this tolerance. The W20T  treatment induced an improvement of cell hydraulic status and photosynthetic pigment levels while reducing membrane disruption and proline accumulation.
           
    These responses could suggest a physiological stability and  an efficiency of the photosynthetic enzyme system such as Rubisco activity and protein content relative to maintain optimal physiological functioning under stress (Wang et al., 2026). 

    CONCLUSION

    This study has demonstrated that diluted seawater (DSW) can serve as a viable irrigation source in agriculture, exhibiting minimal negative effects on plant development. Irrigation of wheat with DSW (at 10%, 20% and 40%) resulted in low concentrations of minerals beneficial for germination, positively correlated with carbonate ions. Additionally, incorporating phosphate into DSW enhanced salinity tolerance and was correlated with better plant growth and tolerance particularly with 0.9% of phosphate. Thus, the use of diluted seawater enriched with optimized doses of phosphate could represent a promising approach for improving crop yields.  

    ACKNOWLEDGEMENT

    The present study was supported by no external funding.
     
    Disclaimers
     
    The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.
     
    Author contributions
     
    Author Hamida Sadji-Ait Kaci: Conceptualization, methodology, investigation, writing original draft. Author Assia Chaker-Haddadj, Farida Taguett and Rafika Ghanai:  Validation, visualization, Author Massakib Bekkaye and Nabi: statistical analysis and data curation, Massakib Bekkaye and Amina Bekkouche: Supervision and visualization. All authors have agreed to the published version of the manuscript.

    CONFLICT OF INTEREST

    The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

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