Promoting Tomato Resilience: Effects of Ascorbic Acid and Sulphur-Treated Biochar in Saline and Non-Saline Cultivation Environments

doi:10.21203/rs.3.rs-4922115/v1

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Promoting Tomato Resilience: Effects of Ascorbic Acid and Sulphur-Treated Biochar in Saline and Non-Saline Cultivation Environments

https://doi.org/10.21203/rs.3.rs-4922115/v1

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The resilience of tomato plants under different cultivation environments, particularly saline and non-saline conditions, was investigated through the application of various treatments, including 0.5% Ascorbic Acid (AsA) and 1% Sulphur-treated Biochar (BS). The study evaluated parameters such as fruit length, diameter, yield per plant and per pot, Total Soluble Solids (TSS) content, chlorophyll content, electrolyte leakage, enzyme activities (Superoxide Dismutase - SOD, Peroxidase - POD, Catalase - CAT), and nutrient content (Nitrogen - N%, Phosphorus - P%, Potassium - K%). Under saline conditions, significant enhancements were observed in fruit characteristics and yield metrics with the application of AsA and BS individually, with the combined treatment yielding the most substantial improvements. Notably, AsA and BS treatments exhibited varying effects on TSS levels, chlorophyll content, electrolyte leakage, and enzyme activities, with the combination treatment consistently demonstrating superior outcomes. Additionally, nutrient content analysis revealed notable increases, particularly under non-saline conditions, with the combined treatment showcasing the most significant enhancements. Overall, the study underscores the potential of AsA and BS treatments in promoting tomato resilience, offering insights into their synergistic effects on multiple physiological and biochemical parameters crucial for plant growth and productivity.

Tomatoes

Salinity

ascorbic acid

Sulphur treated biochar

antioxidants

macronutrients

Tomato (Solanum lycopersicum) is one of the most economically significant vegetable crops worldwide, valued for its nutritional content, culinary versatility, and widespread culinary use. However, tomato cultivation faces numerous challenges, including abiotic stresses such as salinity, which can significantly reduce yields and quality [1, 2]. Salinity stress, resulting from the accumulation of soluble salts in the soil, disrupts plant water balance, ion uptake, and cellular metabolism, leading to osmotic stress, ion toxicity, and oxidative damage. Saline soil is a significant element that restricts crop production. By 2050, it is anticipated that 50% of the world's arable land will be affected by salt stress[3]. Sodium chloride stress, common salt stress, significantly harm crop growth through osmotic stress and particular ion toxicities. Salt stress can affect crops at every growth stage, from germination to maturity. Germination and early seedling growth phases are more salt-sensitive in most plant species than other growth stages [4].

In recent years, there has been growing interest in developing sustainable agricultural practices to enhance crop resilience and productivity under adverse environmental conditions. Among the various strategies investigated, the application of ascorbic acid (vitamin C) and sulfur-treated biochar has shown promising results in improving plant tolerance to salinity stress and enhancing overall growth and yield. Ascorbic acid acts as a potent antioxidant, scavenging reactive oxygen species and protecting cellular components from oxidative damage [4]. At the same time, sulfur-treated biochar improves soil structure, nutrient availability, and microbial activity, thereby promoting plant growth and stress tolerance [5]. This study aims to investigate the effects of ascorbic acid and sulfur-treated biochar on tomato resilience under both saline and non-saline cultivation environments. By elucidating the physiological and biochemical mechanisms underlying the interactions between these treatments and salinity stress, we seek to provide insights into potential strategies for enhancing tomato productivity and sustainability in saline-affected regions. Additionally, we aim to assess the feasibility and economic viability of implementing these strategies in commercial tomato production systems, considering factors such as cost-effectiveness, environmental impact, and long-term sustainability.

The study aims to evaluate the practicality and efficiency of using saline water for irrigating tomato plants (Solanum lycopersicum L.). Tomatoes are fairly resilient to salt stress. A recent analysis has examined the effects of salinity on various aspects of tomato plants, including morphology, physiology, biochemistry, yield, fruit quality, and gene expression [5, 6]. To assess the possible use of biochar to mitigate salinity-related difficulties in tomato cultivation. The study evaluated the impact of salt stress alone or combined with biochar added to the growth medium on plant growth, chlorophyll content, antioxidant response activation, and nutritional status. Through a comprehensive analysis of the effects of ascorbic acid and sulfur-treated biochar on tomato growth, physiology, and yield, this study aims to contribute to the development of innovative and environmentally friendly approaches to mitigate the adverse effects of salinity stress on tomato cultivation. Promoting tomato resilience through targeted interventions can enhance food security, livelihoods, and agricultural sustainability in regions affected by salinity and other abiotic stresses[7, 8].

Experimental site

The evaluation was carried out under pot experiment in the research area of the Faculty of Agricultural Sciences and Technology, Bahauddin Zakariya University Multan (Punjab) Pakistan, which is situated at geographical coordinates 30°15′49″N &71°30′35″E. The experimental design applied completely randomized design (CRD) with four replications for the study execution year, i.e., 2023. The physiochemical properties of soil and irrigation water were given in Table 1.

Table 1

Pre-experimental soil and irrigation characteristics
Soil	Values	References	Irrigation	Values	References ences
pH	819	[19]	pH	7.26	[20]
ECe (dS/m)	2.42	[21]	EC (µS/cm)	387
SOM (%)	0.60	[22]	Carbonates (meq./L)	0.00
TN (%)	0.03	[23]	Bicarbonates (meq./L)	4.69
AP (µg/g)	6.27	[24]	Chloride (meq./L)	0.05
EK (µg/g)	127	[25]	Ca + Mg (meq./L)	2.69
ENa (µg/g)	116	[26]	Sodium (mg/L)	106
Texture	Clay Loam	[27]
TN = Total Nitrogen; AP = Available Phosphorus; EK = Extractable Potassium; ENa = Extractable Sodium

Treatment Application

To investigate the effects of Ascorbic Acid (AsA) and Sulphur-treated Biochar (BS) on tomato resilience under saline (6.52 dS/m) and non-saline (1.38 dS/m) growth conditions. Ascorbic acid (AsA) solutions at concentrations of 0.5% were prepared by dissolving Ascorbic Acid powder in distilled water. Biochar treated with sulphur at a concentration of 1% was utilized in the experiment.

Pot preparation and sowing

Tomato seeds were sown in plastic pots of capacity 20kg filled with a standardized soil mixture. Before sowing, the saline soil pots were mixed with the 1% BS, and upon germination and seedling establishment, the tomato plants were subjected to 0.5% AsA spray.

Collecting, sterilization and sowing of seeds

Tomato seeds of FM-9 variety (procured from an approved seed merchant licensed by the Government of Punjab, Pakistan) were used in this study. The selected seeds were subjected to a rigorous surface sterilization protocol (applied before sowing). The seeds were rinsed three times with 95% ethanol after exposure to the 5% sodium hypochlorite solution. The seeds were then further washed four times with sterilized deionized water to remove any residues of the sterilizing agents. Twenty seeds were sown per pot filled with 5 kg soil. After germination, thinning was achieved at 10 seedlings per pot (Ahmad et al., 2014).

Harvesting and data Collection

A total of 2 harvestings were performed. The first harvesting includes the collection of fresh leaves after 50 days of transplantation (at vegetative stage) for the analysis of chlorophyll contents, proline, antioxidants and other biochemical analysis [28]. However, after 85 days of transplantation tomatoes picking started. For assessment of yield total 8 picks were taken [28]. For the assessment of yield data, a total of 8 picks were taken. The study encompassed the examination of various parameters, including the total plant dry weight (g/plant), plant height (cm), the number of primary branches per plant, fruit length (cm), fruit girth (cm), fruit yield (kg/plant), and the assessment of chlorophyll and proline content. Fruit length and diameter were measured by Vernier caliper and fruit yield per pot were recorded at harvest. Total Soluble Solids (TSS) content was measured using a refractometer. Chlorophyll content was determined using a SPAD meter. Enzyme activities (Superoxide Dismutase - SOD, Peroxidase - POD, Catalase - CAT) were quantified using spectrophotometric assays.

Irrigation

Irrigation in each pot was monitored and adjusted as needed with electrical meters (ADVANCED™; 4 in 1 Soil Meter, China). Daily observations were done to ensure that the moisture level was maintained at wet; on the instrument scale, this corresponds to 70% of field capacity in soil.

Electrolyte leakage

Electrolyte leakage of plants was determined by incubating one leaf in 20 ml ddH2O in darkness for hours). The samples were then vortexed, and initial electrical conductivity was measured with the help of a conductivity meter. Samples were then autoclaved for 15 min at 60 o C. The samples were then cooled to room temperature, and their conductivity was determined. The electrolyte leakage (µS/cm) was calculated according to the following equation: Lutts et al. 1996)

$$\:\:\left(EL\%\right)=Initail\:electrical\:conductivity÷final\:electrical\:conductivity\:\times\:100$$

Macronutrient legislation

Nitrogen, phosphorous and potassium in plant samples were determined as explained by the ICARDA manual 3rd edition (Ryan et al., 2001).

Statistical Analysis

The data were analyzed using conventional statistical techniques, namely the two-way ANOVA, to determine treatment significance. The Tukey test was applied for the paired comparisons, with a significance level of p ≤ 0.05. OriginPro Software was used for the Pearson correlations [36]

Fruit length (cm), Fruit diameter, Fruit plant^− 1 and Yield Pot^–1 (kg)

Under saline conditions, treatment with 0.5% Ascorbic Acid (AsA) resulted in an 11.05% increase in fruit length compared to the control (CK), while 1% Sulphur-treated Biochar (BS) led to a 12.63% increase. The combined treatment of 0.5% AsA and 1% BS exhibited the most significant enhancement, showing a 26.73% increase in fruit length relative to CK (Fig. 1A). In contrast, under non-saline conditions, 0.5% AsA led to a 2.86% increase, 1% BS resulted in a 7.38% increase, and the combined treatment showed a 26.67% increase compared to CK. For fruit diameter, under saline conditions, 0.5% AsA resulted in a 12.43% increase, 1% BS led to an 18.10% increase, and the combined treatment achieved a 24.59% increase compared to CK. In non-saline conditions, 0.5% AsA led to a 3.45% increase, 1% BS resulted in a 10.15% increase, and the combined treatment showed a 13.30% increase in fruit diameter compared to CK (Fig. 1B).

In terms of fruit yield per plant, under saline conditions, 0.5% AsA increased yield by 21.7%, and 1% BS resulted in a 33.2% increase. The combination of 0.5% AsA and 1% BS demonstrated the most pronounced effect, with a 44.2% increase in yield compared to CK. Under non-saline conditions, 0.5% AsA led to a 23.8% increase, 1% BS resulted in a 35.5% increase, and the combined treatment achieved the highest enhancement with a 53.3% increase compared to CK (Fig. 1C).

For yield per pot, under saline conditions, 0.5% AsA resulted in an 18.4% increase, and 1% BS led to a 24.5% increase. The combined treatment of 0.5% AsA and 1% BS showed the most substantial rise, with a 47.4% increase in yield per pot compared to CK. Under non-saline conditions, the addition of 0.5% AsA resulted in a marginal increase of 3.2%, while 1% BS showed no significant difference. The combined treatment of 0.5% AsA and 1% BS did not provide additional benefits compared to the individual treatments (Fig. 1D).

Total soluble solids, Chlorophyll content, Electrolyte leakage and SOD

Examining the Total Soluble Solids (TSS) content, measured in degrees Brix (°Brix), of tomato fruits under different growth conditions and treatments revealed distinct patterns. Under saline conditions, the control (CK) exhibited a TSS level of 8.66 °Brix. Treatment with 0.5% Ascorbic Acid (AsA) resulted in a slight decrease to 8.25 °Brix, while 1% Sulfur-treated Biochar (BS) led to a decrease to 8.34 °Brix. The combined treatment of 0.5% AsA and 1% BS showed the most significant reduction, with a TSS level of 7.56 °Brix compared to CK(Fig. 2A). In contrast, under non-saline conditions, CK displayed a TSS level of 8.34 °Brix. The application of 0.5% AsA and 1% BS resulted in minimal decreases to 8.30 °Brix and 8.24 °Brix, respectively. The combined treatment of 0.5% AsA and 1% BS exhibited a more substantial decrease, with a TSS level of 7.35 °Brix compared to CK.

For chlorophyll content, measured in SPAD units, under saline conditions, CK exhibited a chlorophyll content of 54.48 SPAD units. Application of 0.5% AsA and 1% BS resulted in slight decreases to 54.24 SPAD units and 55.27 SPAD units, respectively. Remarkably, the combined treatment of 0.5% AsA with 1% BS showed the most significant increase, measuring 62.21 SPAD units(Fig. 2B). In non-saline conditions, CK displayed a chlorophyll content of 56.01 SPAD units. Application of 0.5% AsA and 1% BS resulted in relatively minor changes to 55.71 SPAD units and 58.45 SPAD units, respectively. The combined treatment of 0.5% AsA with 1% BS showed a remarkable increase, measuring 65.30 SPAD units compared to CK.

Analysis of electrolyte leakage, represented as a percentage, under saline conditions, CK exhibited an electrolyte leakage of 21.41%. Treatment with 0.5% AsA decreased electrolyte leakage to 19.01%, while 1% BS resulted in a decrease to 17.45%. The combined treatment of 0.5% AsA and 1% BS showed the most significant reduction, with an electrolyte leakage of 15.35% compared to CK(Fig. 2C). In non-saline conditions, CK displayed an electrolyte leakage of 19.52%. Application of 0.5% AsA decreased leakage to 18.19%, while 1% BS led to a reduction to 13.38%.

For Superoxide Dismutase (SOD) activity, measured in units per milligram of protein, under saline conditions, CK exhibited an SOD activity of 23.71 units/mg protein. Treatment with 0.5% AsA decreased SOD activity to 21.56 units/mg protein, and 1% BS resulted in a decrease to 20.45 units/mg protein. The combined treatment of 0.5% AsA with 1% BS showed the most significant reduction, with an SOD activity of 19.41 units/mg protein(Fig. 2D). In non-saline conditions, CK displayed an SOD activity of 22.19 units/mg protein. Application of 0.5% AsA decreased SOD activity to 20.34 units/mg protein, and 1% BS resulted in a reduction to 19.38 units/mg protein. The combined treatment of 0.5% AsA with 1% BS exhibited the most substantial reduction, measuring 17.45 units/mg protein compared to CK.

Peroxidase(POD),CAT, Nitrogen(%),Phosphorus(%)

Investigation into Peroxidase (POD) activity, measured in units per milligram of protein, in tomato plants grown under varying conditions and subjected to different treatments revealed notable trends. Under saline conditions, the control (CK) exhibited a POD activity of 12.63 units/mg protein. Application of 0.5% Ascorbic Acid (AsA) led to a decrease in POD activity to 10.27 units/mg protein. The application of 1% Sulfur-treated Biochar (BS) resulted in a further reduction, measuring 8.34 units/mg protein. Remarkably, the combination treatment of 0.5% AsA with 1% BS also exhibited a POD activity of 8.34 units/mg protein, indicating a similar effect to the standalone BS treatment (Fig. 3A). In contrast, under non-saline conditions, CK displayed a POD activity of 10.67 units/mg protein. Application of 0.5% AsA led to a slight decrease to 9.36 units/mg protein, while 1% BS resulted in a further reduction to 8.32 units/mg protein. Notably, the combination treatment of 0.5% AsA with 1% BS exhibited the most substantial decrease in POD activity, measuring 6.65 units/mg protein compared to CK.

Investigation into Catalase (CAT) activity, measured in units per milligram of protein, revealed significant trends. Under saline conditions, CK exhibited a CAT activity of 6.41 units/mg protein. Application of 0.5% AsA resulted in a slight decrease to 6.09 units/mg protein. Similarly, the application of 1% BS led to a further reduction to 5.75 units/mg protein. Remarkably, the combination treatment of 0.5% AsA with 1% BS displayed the lowest CAT activity, measuring 5.16 units/mg protein (Fig. 3B). In contrast, under non-saline conditions, CK group exhibited a CAT activity of 6.19 units/mg protein. Application of 0.5% AsA led to a slight decrease to 5.90 units/mg protein. The application of 1% BS resulted in a further reduction to 4.72 units/mg protein. Notably, the combination treatment of 0.5% AsA with 1% BS exhibited the most substantial decrease, measuring 3.27 units/mg protein compared to CK.

In examining the Nitrogen content (N%) of tomato plants, notable trends emerged under both saline and non-saline conditions. Under saline conditions, the application of 0.5% AsA led to a modest increase in N% by approximately 7.48% compared to CK, while the application of 1% BS resulted in a more substantial rise of about 19.95%. Impressively, the combination treatment of 0.5% AsA with 1% BS exhibited the highest increase in N% by approximately 25.06% compared to CK (Fig. 3C). Conversely, under non-saline conditions, the individual application of 0.5% AsA led to a considerable increase in N% by approximately 12.07% compared to CK, while the application of 1% BS resulted in a more pronounced rise of about 32.61%. Remarkably, the combination treatment of 0.5% AsA with 1% BS displayed the most significant enhancement in N% by approximately 39.78% compared to CK.

In analyzing the Phosphorus content (P%) of tomato plants, distinct patterns emerged under both saline and non-saline conditions. Under saline conditions, the application of 0.5% AsA led to a moderate increase in P by approximately 24% compared to CK, while the application of 1% BS resulted in a slightly higher rise of about 32%. Notably, the combination treatment of 0.5% AsA with 1% BS displayed the highest increase in P by approximately 36% compared to CK (Fig. 3D). In contrast, under non-saline conditions, the individual application of 0.5% AsA resulted in a noticeable increase in P by approximately 6% compared to CK, while the application of 1% BS led to a more pronounced rise of about 15%. Remarkably, the combination treatment of 0.5% AsA with 1% BS exhibited the most significant enhancement in P by approximately 18% compared to CK.

Potassium

Analyzing tomato plants' potassium content (K%) under both saline and non-saline conditions relative to the CK group reveals significant variations. Under saline conditions, the application of 0.5% AsA led to a notable increase in K by approximately 27.5% compared to the CK, while the application of 1% BS resulted in a more substantial rise of about 96.88%. Remarkably, the combination treatment of 0.5% AsA with 1% BS exhibited the highest increase in K by approximately 113.75% compared to the CK. Conversely, under non-saline conditions, the individual application of 0.5% AsA resulted in a decrease in K by approximately 6.20% compared to the CK, while the application of 1% BS led to a notable increase of about 34.88%. Notably, the combination treatment of 0.5% AsA with 1% BS exhibited the most substantial enhancement in K by approximately 43.02% compared to the CK (Fig. 4).

The study examined how different doses of ascorbic acid (AsA) mixed with biochar affect the emergence, early seedling growth, and physiological characteristics of tomato plants under salinity stress. We discovered that the simultaneous use of AsA and BC at the right concentration improved salt tolerance in tomato plant by boosting emergence, seedling growth, and physiological functions under salt stress. Seedling growth, namely root and shoot development, is typically robust and establishes a strong basis for achieving optimal crop density, particularly in high salinity conditions [9, 10]. Insufficient seedling emergence is a primary factor that hinders crop establishment in saline conditions.

Dissolved salts in the soil solution near plant roots might hinder plant growth. The osmotic impact reduces the water uptake of plants, leading to a decrease in the water potential of the plant's leaves and tissues[11, 12]. An elevated accumulation of salts in the plant's tissues will hinder the plant's growth and productivity by disrupting crucial processes such as germination, photosynthesis, nutrient balance, and redox balance. This will restrict the plant from achieving its maximum growth potential [13, 14].

Research has demonstrated that biochar can trap sodium ions, restricting plant roots' absorption. Our study shows that raising the biochar quantity to 2 tons per hectare successfully counteracts the decrease in fruit yield due to salinity and reduces the sodium accumulation in the roots and fruit. Xiao et al. (2022) found a reduction of salt levels in wheat grains grown in low-fertile soil affected by salinity in the Yellow River Delta by adding 12 tons of biochar per hectare [15, 16]. Increasing the amount of biochar added to a tomato plant could enhance its ability to adapt to saline water by decreasing the absorption of sodium into the plant tissue. However, this aspect was not explored in this study. This theory was never tested. Extreme caution is necessary when adding biochar to clay soil as a high amount could upset the balance between liquid and gaseous phases, reducing the water available in the rhizosphere[17, 49].

The combined treatment of 0.5% AsA with 1% BS exhibited the most significant enhancement, showing a 26.73% increase compared to the CK. In contrast, the percent increase in non-saline conditions compared to the CK group was lower for all treatments. Treatment with 0.5% AsA resulted in a modest 3.45% increase, while 1% BS led to a more noticeable 10.15% increase in fruit diameter. Interestingly, the application of 0.5% AsA and 1% BS showed slight decreases in TSS levels compared to the CK, with values of 8.25 °Brix and 8.34 °Brix, respectively. Notably, the combination treatment of 0.5% AsA with 1% BS resulted in a more pronounced decrease, with a TSS level of 7.56 °Brix. In contrast, the CK displayed a TSS level of 8.34 °Brix under non-saline conditions.

Superoxide dismutase (SOD) is a primary scavenger of O2 – and its enzymatic activity leads to the formation of H2O2 and O2. H₂O₂ is eliminated by peroxidases (POD) or catalases (CAT) into water and oxygen [18]. Salt stress led to a notable decrease in the activity of POD, SOD, and CAT enzymes in our research. Another study confirmed that under salt stress, the activities of POD, SOD, and CAT were decreased, as observed by [20, 22, 23, 50]. Reduced enzyme activity in stressed plants may have triggered the accumulation of H₂O₂ in plant cells under stress, ultimately leading to damage in biological systems [19]. Contrary findings were presented by [20, 21], indicating that salt stress led to elevated activity of POD, SOD, and CAT. An elevation in antioxidant enzymes during salt stress may indicate a rise in reactive oxygen species (ROS) and an enhancement of a defense mechanism to reduce oxidative damage caused by stress in plants. 200 millimolar NaCl POD, SOD, and CAT showed the highest activity at a high level of AsA and the lowest activity at 0 .5%level of AsA. Notably, the combination treatment of 0.5% AsA with 1% BS showed the most significant reduction in SOD activity, recording at 19.41 units/mg protein [36, 37, 38]. Biochar application in salty soil can enhance seedling growth physiological and biochemical processes through the controlled release, retention, or immobilization of nutrients due to its surface features, such as cation exchange and anion exchange capacity [21, 22, 48]. The study revealed that the use of BC elevated the activity of antioxidant enzymes, including CAT, SOD, and POD, in sorghum seedlings. Many researchers have found that the administration of BC improves stress tolerance by enhancing the activity of POD, SOD, and CAT. The BC treatment can control the production of antioxidant enzymes in plants, improving their tolerance to salt stress. However, other studies have shown that the application of BC decreased the activity of the POD, POD, and CAT enzymes in salt conditions, the combination treatment of 0.5% AsA with 1% BS displayed a CAT activity of 5.16 units/mg protein The decrease in antioxidant enzyme activity may be linked to the ability of biochar to immobilize heavy metals, resulting in reduced metal translocation into plant tissues [23, 25, 46, 47]. The most significant levels of POD, SOD, and CAT activity were found when using a 4% BC treatment. The activity of antioxidant enzymes rose as the concentration of ASA increased under salt-stress conditions [24, 39, 40, 41]. Our investigation found that external ASA might enhance the activity of POD, SOD, and CAT in the presence of NaCl. ASA was found to enhance the activities of POD, SOD, and CAT, which are essential for the anti-oxidative defense needed by plants under saline conditions [26, 27, 28, 29]. Enhanced activity of antioxidant enzymes may result from the regenerative properties of ascorbate, which aids in neutralizing reactive forms of molecular oxygen by direct action or enzymatic processes [43, 44, 45].

The findings of the study are expected to provide valuable insights into the effectiveness of ascorbic acid and sulphur-treated biochar in promoting tomato resilience under saline stress. Positive outcomes, such as improved plant growth, yield, and physiological parameters, would indicate the potential utility of these interventions in mitigating the adverse effects of saline soil conditions on tomato cultivation(Fig. 5). Moreover, the study may highlight the importance of tailored management practices for different environmental conditions to enhance crop resilience and ensure food security [36, 41, 42].

In addition, the acid-modified biochar had a stronger beneficial impact on the plant's dry matter quantity and yield at the same addition level compared to regular biochar in this experiment. ascorbic acid was likely used to enhance the physicochemical qualities of biochar by removing metals and contaminants from its surface and pores, hence increasing its pore structure and specific surface area [30, 31, 35]. Acid-modified biochar has enhanced nutrient absorption capacity due to its larger and more numerous internal pores, leading to gradual nutrient release throughout plant growth and minimizing soil nutrient depletion. Biochar is typically alkaline, however treating it with phosphoric acid lowers its pH [32, 33, 34]. Acid-modified biochar added to saline soils neutralizes alkalinity and regulates soil pH more effectively.

Pearson correlation analysis

The pearson correlation analysis presented a relatively strong association between multiple variables in the dataset. Plant dry weight showed a very strong positive correlation of 1 with plant height, meaning increased plant height led to a significant increase in plant dry weight. Similarly, No. of primary branches/plant, Fruit length, fruit girth, fruit yield, chlorophyll a, and chlorophyll b indicated positive relations with almost 1 value. However, factors such as EL, %, TSS, MDA, POD, CAT and SOD had a strong negative correlation, which can be deemed as a negative relation with other variables. But surprisingly, leaves N, leaves P, leaves K, and leaves Na presented correlations. Leaves N and leaves P expressed almost positive correlation of 0.985, and leaves K and leaves Na had positive and negative correlations of around 0.997 and − 0.986, which suggests the association among the following variables

The research findings highlight the significant impact of different treatments on tomato crop growth under both saline and non-saline conditions. Notably, the application of 0.5% Ascorbic Acid (AsA) and 1% Sulphur-treated Biochar (BS) resulted in enhancements across various parameters, including fruit morphology, yield, and physiological responses. Under saline conditions, these treatments led to substantial increases in fruit length, diameter, and yield per plant and per pot compared to the control group. Impressively, the combined treatment of 0.5% AsA with 1% BS consistently outperformed individual treatments, demonstrating the most significant improvements in fruit quality and yield metrics. Moreover, analyses of Total Soluble Solids (TSS) content, chlorophyll levels, and enzymatic activities revealed nuanced responses to the treatments, with the combination treatment often eliciting the most favorable outcomes, particularly in mitigating salinity-induced stress. Furthermore, investigations into nutrient content showcased the potential of combined AsA and BS application to enhance nitrogen, phosphorus, and potassium uptake, contributing to improved plant nutrition and growth. Overall, these findings underscore the efficacy of integrated approaches involving antioxidant supplementation and biochar application in promoting tomato crop productivity and resilience to environmental challenges, offering valuable insights for sustainable agricultural practices. Further research is warranted to elucidate the underlying mechanisms and optimize the application protocols for maximizing the benefits of these treatments in agricultural systems.

Author Contributions

M.I.: Conceptualisation, methodology, software, formal analysis, investigation, data curation, visualisation, writing—original draft preparation. AM.: Conceptualisation, methodology, investigation, resources, supervision, project administration, writing—review and editing. A.A.A.: Conceptualisation, methodology, investigation, resources, writing—review and editing. A.M.G. S. Danish: methodology, resources, writing—review and editing. E.M.; M.T.; M.F.K.M.: A.R.: writing—review and editing. All authors have read and agreed to the published version of the manuscript.

Acknowledgements:
The authors express their gratitude to Princess Nourah bint Abdulrahman University Researchers Supporting Project number (PNURSP2024R93), Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia.

Funding

This research was funded by Princess Nourah bint Abdulrahman University Researchers Supporting Project number (PNURSP2024R93), Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia.

Data Availability Statement

The data presented in this study are available on request from the corresponding author.

Conflicts of Interest

The authors declare no conflicts of interest.

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You are reading this latest preprint version

Promoting Tomato Resilience: Effects of Ascorbic Acid and Sulphur-Treated Biochar in Saline and Non-Saline Cultivation Environments

Status:

Version 1

Abstract

Figures

Introduction

Materials and methods

Experimental site

Treatment Application

Pot preparation and sowing

Collecting, sterilization and sowing of seeds

Harvesting and data Collection

Irrigation

Electrolyte leakage

Macronutrient legislation

Statistical Analysis

Results

Fruit length (cm), Fruit diameter, Fruit plant^− 1 and Yield Pot^–1 (kg)

Total soluble solids, Chlorophyll content, Electrolyte leakage and SOD

Peroxidase(POD),CAT, Nitrogen(%),Phosphorus(%)

Potassium

Discussion

Pearson correlation analysis

Conclusion

Declarations

References

Additional Declarations

Status:

Version 1

Promoting Tomato Resilience: Effects of Ascorbic Acid and Sulphur-Treated Biochar in Saline and Non-Saline Cultivation Environments

Status:

Version 1

Abstract

Figures

Introduction

Materials and methods

Experimental site

Treatment Application

Pot preparation and sowing

Collecting, sterilization and sowing of seeds

Harvesting and data Collection

Irrigation

Electrolyte leakage

Macronutrient legislation

Statistical Analysis

Results

Fruit length (cm), Fruit diameter, Fruit plant− 1 and Yield Pot–1 (kg)

Total soluble solids, Chlorophyll content, Electrolyte leakage and SOD

Peroxidase(POD),CAT, Nitrogen(%),Phosphorus(%)

Potassium

Discussion

Pearson correlation analysis

Conclusion

Declarations

References

Additional Declarations

Status:

Version 1

Fruit length (cm), Fruit diameter, Fruit plant^− 1 and Yield Pot^–1 (kg)