Synergistic  phytoremediation effect by Sorghum bicolor, citric acid, biochar, and Vermiwash amendment for the remediation of a mine-contaminated soil

Synergistic phytoremediation effect by Sorghum bicolor, citric acid, biochar, and Vermiwash amendment for the remediation of a mine-contaminated soil | 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 Research Article Synergistic phytoremediation effect by Sorghum bicolor, citric acid, biochar, and Vermiwash amendment for the remediation of a mine-contaminated soil Hanan E. Osman, Ruwaydah S. Fadhlallah, Mohamed H.E. El-Morsy This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3445202/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 5 You are reading this latest preprint version Abstract Phytoremediation technology is an eco-friendly technology for the treatment of a polluted environment. In contrast, it has been demonstrated that both natural and synthetic amendments can enhance the process of phytoremediation of heavy metals (HMs) from polluted soils through the utilization of bioenergy crops. This work assessed the synergistic impact of two tested biochar (BC) from data palm (B1) and Prosopis (B2)/ citric acid (CA) and Vermiwash (VW) to enhance phytoremediation of tested HMs (Zn, Pb, Cd, Ni, Cu, Mn, and Fe) from mine-contaminated soil by Sorghum (Sorghum bicolor). The BC and CA amendments alone and combined with VW significantly augmented the proliferation and survival of sorghum grown in mine-contaminated soil. Considering the individual and combined applications of VW and BC, the influence on plant growth followed this order: K > VW > B2 > B1 > B1 + VW > B2 + VW > CA > CA + VW. Applying tested BC/ CA and VW significantly increased chlorophyll compared to unamended soil. The outcomes revealed a substantial elevation in HM absorption in both shoot and root (p ≤ 0.05) with all tested treatments compared to the untreated soil (K). The combined application of CA and VW resulted in the highest uptake of HMs in both the root as well as the shoot. This study highlights the efficacy of combining CA/BC with VW as a more viable option for remediating mine-contaminated soil compared to individual amendments. Mahad AD'Dahab mine contaminated soil biochar cirtic acid phytoremediation sorghum vermiwash Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction As an integral component of the Earth’s ecosystem, it serves a crucial function in sustaining the quality of life for humans as well as the survival of various macro and microorganisms. Unfortunately, numerous detrimental substances are being released into the environment (as liquid and solid waste) by mining and industrial operations, leading to significant pollution in the surrounding areas (Baldauf et al. 2001 ). In addition, m activities have led to the alteration of the natural landscape, the contamination of water and soil resources, and soil compaction through the discharge of numerous hazardous substances ( Rodriguez-Franco and Page-Dumroes 2021 ). Among these harmful substances, HMs are frequently found near waste disposal sites, causing substantial water as well as soil contamination ( Granero and Domingo 2002; Nadal et al. 2005; Ferreira et al., 2021 ). Such contaminated soil is unsuitable for agricultural practices, as it can accumulate HMs in edible crops, rendering them unfit for consumption ( Chen et al. 2015; Jia et al. 2018 ). Therefore, restoring these polluted sites using economically viable techniques is imperative before they can be utilized for agricultural purposes, ensuring their suitability for productive farming activities ( Oladoye et al., 2022; Quronfulah et al., 2023) . Traditional soil remediation methods involve destructive practices like excavation, landfilling, and acid washing. However, there is a growing interest in developing and utilizing cost-effective, in-situ, and environmentally friendly technologies for soil decontamination and metal recovery. Phytoremediation methods, which involve the utilization of particular plant species for the purposes of immobilizing contaminants (phytostabilization), extracting them from the environment (phytoextraction), or converting them into less detrimental forms, have garnered significant attention ( Prasad, 2003; Vangronsveld et al., 2009; Alegbeleye et al., 2017 ; Oladoye et al., 2022 ). In order to render contaminated lands suitable for agricultural activities, they must be restored using economically feasible techniques before being used for agricultural production. Stabilization of HMs-contaminated soil in situ has been deemed viable, cost-effective, and simple to implement. Several additives have been used to stabilize in situ, including chelating agents, organic adsorbents, compost, biosolids, and activated carbon ( Rafique et al., 2020 ). Over the past few years, various phytoremediation approaches have been employed, with processes involving organic amendments gaining increased attention ( Wiszniewska et al., 2016 ). These approaches encompass the use of organic acids (decreased molecular weight) ( Duarte et al., 2011; Chen et al., 2020; Helaoui et al., 2023; Wang et al., 2023 ), the introduction of plant growth-promoting substances (e.g., VW through bioaugmentation) ( Paredes-Paliz et al., 2018 ), and in recent times, the implementation of BC ( Ghosh and Maiti, 2021; Wang et al., 2023 ). Biochar (BC), a product of biomass pyrolysis, is used in various environmental applications because it is a cost-effective, carbonaceous material with marked efficacy and versatility and a large surface area with numerous active sites, particularly as a soil amendment for HM remediation ( He et al., 2019; Ghosh and Maiti, 2021 ). The selection of raw materials utilized in the manufacturing of biochar and the specific conditions applied during the pyrolysis process are crucial factors that have a significant influence on the characteristics of biochar-metal binding and its impacts on soil ( Qi et al., 2017; Fan et al., 2020; Harindintwali et al., 2020 ). Another viable approach to promote the solubility of potential soil’s toxic metals is adding chelating agents. Chelates like organic acids (of low molecular weight) boost plants’ uptake of potentially toxic metals (Anning and Akoto, 2018 ; Yin et al., 2015 ). CA has been utilized as an amendment to enhance the elimination of different metals owing to its environmentally sustainable characteristics, facile biodegradability, and non-toxic characteristics (Anwer et al., 2012 ; Al Mahmud et al., 2018 ). Numerous studies have proven that CA significantly contributes to phytoremediation enhancement in multiple metals by improving metal solubility and mobilization. In addition, CA contributes to mitigating metal-induced oxidative stress by enhancing the activity of enzymes in antioxidative defense systems (Al Mahmud et al., 2018 ). The primary objectives of all these organic amendments are twofold: i) to enhance elements’ phytostabilization or photo-availability for extraction, and ii) to promote plant resistance as well as tolerance to the uptake of potentially toxic metals ( Duarte et al., 2011; Ghosh and Maiti, 2021 ). These objectives aim to improve phytoremediation performance regarding element extraction, plant health, and subsequent biomass production. However, there is limited research on using VW and its effects on plants cultivated in contaminated soils. VW, an extract from vermicomposting, is a brown-colored liquid rich in plant growth-promoting hormones, calcium, zinc, potassium, amino acids, vitamins, and nitrogen (Aghamohammadi et al., 2016 ). Numerous studies have been conducted to investigate the potential of VW as a bio-fertilizer and pesticide for augmenting crops’ productivity and growth. The study is designed to achieve the following goals: 1) Investigate the combined impact of CA, BC, and VW on the phytoremediation of mine-contaminated soil. 2) Evaluate how different amendments affect sorghum plants’ growth, biomass, and chlorophyll content. 3) Examine the tested HM accumulation in sorghum plants cultivated in mine-contaminated soil. Materials and Methods Vermiwash preparation and analysis Vermiwash (VW) was produced through the secretion of Eudrillus euginiae earthworms, extracted as the seepage drained from the worm bed, and then preserved in earthen pots (Fig. 1 ), following the established technique described by Ismail (1997) . This collected VW was utilized in its undiluted form, i.e., 100% concentration, the characteristics of the VW (Table 1 ). Table 1 Chemical properties of vermiwash (VW) fertilizer used pH EC Ca Cd Cu Fe Mn Ni Pb Zn P N TOC mg/L % mg/l 7.65 1.78 4.38 nd 0.03 0.25 1.50 0.97 nd nd 1.71 0.522 521.88 The electrical conductivity (EC) as well as chemical reactions (pH) were examined using the methodology outlined by Garg et al. (2006) . Additionally, total organic carbon (TOC) content was assessed following the procedure established by Nelson and Sommers (1982) . Total nitrogen (N) was quantified using the method illustrated by Bremmer and Mulvaney (1982) . Finally, total P, K, Na, and Ca were determined following the procedure described by Bansal and Kapoor ( 2000 ) . Production and characterization of tested BC Date palm ( Phoenix dactylifera ) and Prosopis ( Prosopis juliflora ) were picked, dried, chopped, and then placed (in a container made of airtight stainless-steel) with a 7 cm diameter and a height of 22 cm. Subsequently, they underwent pyrolysis in an electrical muffle furnace at 450°C for 4 hours ( Al-Wabel et al. 2013; Usman et al. 2015 ). Both biochar samples were subsequently pulverized and filtered using a 1.0-mm sieve ( Supplementary Fig. 1 ). The physiochemical properties of these biochar samples are outlined in Table 2 . The EC and pH were determined using an aqueous extract of the BC at a 1:10 ratio. The analysis of dried BC samples was done in terms of their C, H, N, and S content using a CHNS analyzer (Vario EL III, Germany). X-ray diffraction (MAXima X XRD-7000-Shimadzu-Japan) was employed to determine any crystal structures formed in the materials ( Supplementary Figs. 2&3 ). Furthermore, examination of BC surface morphology was done via scanning electron microscopy (SEM) utilizing (JSM-6460 LV-Scanning Microscope) ( Supplementary Figs. 4&5 ). Table 2 Physiochemical properties of used biochar (n = 3) Physiochemical properties pH EC (mS/cm) Chemical composition (%) C H N S C/N ratio Date palm 8.15 3.42 56.20 4.18 0.60 2.11 93.67 Prosopis 7.94 6.17 72.20 3.15 0.60 0.69 120.33 Greenhouse experiment and soil and plant sampling This study collected soil from the surface layer at the Mahad AD’Dahab gold deposit site. This mining location is 380 km northeast of Jeddah, precisely at coordinates 23°30’30” N latitude and 40°51’30” E longitude, in the late Proterozoic Arabian Shield’s west-central region (Fig. 2 ). A controlled pot experiment was conducted at (Umm Al-Qura University, Saudi Arabia) at coordinates 21°18’56.3” N latitude and 39°56’47.8” E longitude from November 2020 to January 2021. The greenhouse setting at temperatures ranging from 20°C to 28°C and a humidity level of 50 ± 4%. A total of 10 sorghum seeds were sown in individual pots containing 4 Kg of mine-contaminated soil. The present study adopted a completely randomized design (CRD), with each treatment replicated six times, as depicted in Table 3 . After 60 and 80 days from sowing, VW was applied as 20 ml/kg soil. Table 3 Treatments in this study. Treatment Description K control VW Vemiwash B1 biochar date palm B1 + VW biochar date palm + vermiwash B2 biochar Prosopis B2 + VW biochar Prosopis + vermiwash CA citric acid CA + VW citric acid + vermiwash In contrast, CA was utilized as a chelating agent at a 1.5 mmol/kg soil rate after 80 days from plantation. Additionally, BC was utilized at a (1.5% / kg soil) rate before cultivation. The moisture levels were adjusted to 60% of the soil’s field capacity. Soil Sampling and Analysis Prior to initiating study procedures, soil samples were obtained from each individual pot. The soil samples underwent the process of air-drying and were subsequently passed through a mesh (with a 2 mm pore size). The main physicochemical characteristics of soil collected from Mahad Adh Dahab were assessed by methods described in Burt (2004) and are outlined in Table 4 . Soil pH and EC were 7.66 and 1.68 ds/m, respectively. Additionally, the soil exhibited a sandy loam texture. Table 4 Soil characteristic, total, and DTPA- extracted Cd, Cu, Fe, Mn, Ni, Pb, and Zn contents in soil collected from Mahad Adh Dahab used in the pot experiment. (mean ± SD, n = 3). Cd Cu Fe Mn Ni Pb Zn pH EC OM Texture mg/Kg Ds/m % Total 9.78 1570.00 790.00 985.00 23.50 491.00 1150.00 7.66 1.68 4.35 Sandy loam SD 1.23 23.50 57.90 27.88 3.55 10.20 25.20 0.99 0.06 0.49 DTPA 2.40 169.52 56.80 78.86 9.25 43.88 116.03 SD 0.17 7.24 4.20 3.48 0.94 4.89 6.84 Acid mixture was utilized to digest the soil samples (HClO 4 : H 2 SO 4 : HNO 3 ; 1:1:5, v/v/v) to evaluate the total HM contents (Zn, Pb, Cd, Ni, Cu, Mn, and Fe) ( USEPA, 1992 ), and their concentrations were identified utilizing an inductively coupled plasma optical emission spectrophotometer (ICPOES; PerkinElmer Optima 4300 DV, USA). Diethylene triamine pentaacetate (DTPA) was used to extract and assess the available tested HM contents in the soil before and after cultivation in each treatment, according to Lindsay and Norvell (1978) . Plant Sampling and Analysis The samples were collected 90 days after the plantation. From each treatment, a suitable number of plant species were randomly selected and combined into three replicate groups for assessing growth criteria. The plants were divided into their root and shoots, and the lengths of both roots and shoots were recorded. Furthermore, the dry and fresh weights (g/ pot) were determined through the weight of different plant parts before and after oven-drying at 65°C until a constant weight was achieved. The photosynthetic pigments in fresh leaves were extracted with 85% acetone, and their concentrations were measured with a spectrophotometer at 662 nm, 644 nm, and 440.5 nm following the procedure Arnon ( 1956 ) . Moreover, the soil analysis procedures assessed seven tested HM contents in plant samples. Data Analysis The bioaccumulation factor (BCF) is a vital instrument for the prediction of HM absorption by plants from the soil. In contrast, HM translocation from roots to shoots is assessed using the translocation factor (TF) in order to determine the efficacy of different treatments in extracting metals from the soil and transporting them to the leaves ( Ghosh and Singh, 2005 ). BCF = C plant / C soil (1) TF = C shoot / C root (2) Where C soil and C plant are the mean of shoot and root and HM concentration (mg/kg) in the soil, respectively. HM uptake (µg/pot) was also conducted, as described by Utmazian et al. (2007) . TU (µg/pot) = HM concentration in roots or shoots dw X d wt of roots or shoots g/pot (3). Statistical analysis The data underwent analysis through a one-way ANOVA using the 22nd version of the IBM-SPSS software. Pearson’s correlation analysis was done to determine the associations between variables and assess their statistical significance with a confidence interval of 95%, and significance was determined at a threshold of p ≤ 0.05. Similarly, multivariate statistics analysis is applied to categorize and evaluate the most suitable method for the removal of HMs. The data can be condensed, unified, and categorized using multivariate approaches to extract meaningful information. The technique can also be used to explain temporal and spatial variations brought on by relationships between seasonality and natural and human influences. The principal component analysis is used to analyze the data collected. The most popular method for converting the original variables into new, uncorrelated variables (axes) or principal components is principal component analysis (PCA). PCA explains data variations clearly ( Pop et al. 2009 ). Results and Discussion Impact of different treatments of DTPA – heavy metal contents HM toxicity in soil is primarily impacted by the availability of metal forms accessible to plants rather than the overall concentration of metals in the soil ( Yin et al., 2016 ). Zn, Pb, Ni, Cd, Mn, Fe, and Cu concentrations before and after cultivating sorghum plants in Mahad AD’Dahab under various treatment conditions are reported in Table 5 . The findings indicate that the use of both tested BC in the soil led to a decrease in DTPA-extractable Zn, Pb, Ni, Cd, Mn, Fe, and Cu (Table 5 ), which is consistent with the outcomes demonstrated by Yousaf et al. (2016) for Cd and Rehman et al. (2016) for Ni. Table 5 DTPA- extracted Cd, Cu, Fe, Mn, Ni, Pb, and Zn contents before and after cultivation for 90 days grown in Mahad AD’Dahab used in the pot experiment. (mean ± SD, n = 3) Treatments Cd Cu Fe Mn Ni Pb Zn Before cultivation K 2.40 ± 0.17 169.52 ± 7.24 56.80 ± 4.20 78.86 ± 3.48 9.25 ± 0.94 43.88 ± 4.89 116.03 ± 6.84 After Cultivation K 2.12 ± 0.28 155.57 ± 5.12 49.70 ± 4.10 69.80 ± 6.22 8.20 ± 1.11 39.12 ± 3.09 103.76 ± 7.15 VW 2.32 ± 0.36 149.38 ± 6.33 51.94 ± 4.57 67.68 ± 5.45 8.71 ± 1.64 41.75 ± 3.89 104.24 ± 6.24 B1 1.80 ± 0.22 137.44 ± 6.71 42.46 ± 3.73 60.73 ± 4.90 6.84 ± 0.96 35.25 ± 3.23 94.72 ± 4.44 B1 + VW 1.64 ± 0.19 140.11 ± 6.18 40.88 ± 2.34 61.13 ± 5.46 6.25 ± 0.94 37.60 ± 3.65 91.39 ± 5.25 B2 1.56 ± 0.14 128.10 ± 4.39 41.23 ± 3.63 59.33 ± 3.84 5.97 ± 0.44 38.47 ± 2.89 93.65 ± 5.84 B2 + VW 1.50 ± 0.16 127.19 ± 7.27 39.13 ± 3.24 57.22 ± 4.90 6.05 ± 0.58 40.01 ± 3.99 96.27 ± 3.61 CA 2.89 ± 0.23 188.04 ± 4.61 65.97 ± 2.84 92.16 ± 5.22 10.97 ± 0.60 49.32 ± 3.43 130.77 ± 5.49 CA + VW 2.80 ± 0.33 183.41 ± 7.59 64.75 ± 3.94 90.41 ± 5.22 10.76 ± 1.16 47.67 ± 3.53 127.81 ± 5.59 Similarly, the outcomes show that the bioavailability of Cd, Cu, Fe, Mn, Ni, Pb, and Zn decreased by 18.92%, 25.00%, 25.25%, 22.99%, 26.05%, 19.67%, and 18.37%, respectively, when date palm (BC1) biochar was applied. In contrast, Prosopis biochar (BC2) reduced 35.00%, 24.43%, 27.41%, 24.77%, 35.46%, 12.33%, and 19.29%, respectively (Table 5 ). These findings confirm that date palm and Prosopis BC have an increased adsorption capacity and the potential to create binding sites for the tested HMs. This finding aligns with previous studies ( Zheng et al., 2016; Bashir et al., 2019 ; Rafique et al., 2021; Ramírez et al., 2022; Wang et., 2022; Alazzaz et al., 2023 ; Li et al., 2023) , Moreover, the strong affinity of HMs to BC may be another reason for reduced metal availability. BC’s larger surface area and numerous contact sites significantly reduce DTPA-extractable Cu, Cd, Fe, Ni, Mn, Zn, and Pb in the soil ( Lu et al., 2017; Naeem et al., 2020 ). Comparable results have been reported in prior research, where BC from various sources effectively reduced Cd, Cu, Fe, Mn, Ni, Pb, and Zn concentrations in mine-contaminated soil ( Xu et al., 2016; Irfan et al., 2021; Alazzaz et al., 2023 ). Furthermore, HM bioavailability in soil is crucial for successful phytoremediation ( Greman et al., 2001 ). In this study, the application of 1.5 mol/Kg soil of CA elevated the bioavailability of Zn, Pb, Ni, Mn, Fe, Cu, and Cd by 11.27%, 10.99%, 15.68%, 14.43%, 13.90%, 9.85%, and 16.96% respectively (Table 5 ). The same results were recorded by Diarra et al. (2021) , who found that applying CA released the most elevated Cu and Ni fractions in the soil, accounting for 75.5% and 79.5%, respectively. This finding is compatible with those of Wuana et al. (2010) , who revealed a significant decline in Cu and Ni levels through batch soil washing with CA. Furthermore, fractionation patterns revealed that CA primarily targets HMs related to the reducible as well as exchangeable fractions. A smaller proportion of HMs bound to the soil organic matter are also targeted by CA ( Wuana et al., 2010 ). Impact of various amendments on morphological traits Plant morphological traits, such as plant height, root length, and plant biomass (both fresh and dry weights), were significantly influenced by various treatments when compared to the untreated treatment (K)(Fig. 3 ). Root and shoot lengths varied between 9.33–12.83 cm and 76.33-88.00 cm, respectively. The sorghum plants cultivated in untreated soil (K) exhibited the shortest root and shoot lengths, while the application of a combination of CA + VW to the soil led to the longest shoot and root lengths. Compared to control (K), the implementation of both tested BCs (B1&B2) significantly enhanced sorghum morphological characteristics. The enhancement in plant height, root length, and fresh and dry biomass was 20.61%, 59.72%, 83.06%, and 54.92% for the B1 treatment and 60.98%, 22.87%, 75.28%, and 61.59% for the B2 treatment. Abbas et al. ( 2017 ) demonstrated that the application of BC substantially impacted the root length, plant height, and biomass of wheat plants when exposed to various Cd treatments compared to untreated conditions. BC soil amendment has been shown to promote the growth of numerous species of plants exposed to metal stress ( Rizwan et al., 2016; Rehman et al., 2016; Younis et al., 2016; Almaroai and Eissa 2020 ; Helaoui et al., 2023 ). In this study, the elevation in these parameters by adding BC may be attributed to increased mineral nutrient availability. Previous research has frequently reported that BC improves plant growth because of improved biological characteristics, increased nutrient content, elevated cation exchange capacity, and pH adjustments ( Paz-Ferreiro et al., 2014; Širi’c et al., 2022 ). Additionally, Irfan et al. (2021) found that applying wheat straw BC in polluted soil significantly enhanced shoot and root lengths and plant biomass in maize plants compared to those grown in contaminated soil without BC. Similar findings were reported by Lu et al. (2014) for edible amaranth, Park et al. (2011) for canola plants, and Ali et al. ( 2017 ) for Indian mustard. In contrast, the application of CA resulted in a significant enhancement of shoot and root lengths, as well as overall plant biomass, in contaminated soil. Fresh and dry weights increased with CA application from 23.5 g and 3.15 g /pot in the K to 48.86 g and 5.85 g/pot. Additionally, sorghum root elongation, plant height, and biomass increased when VW was applied with BCand CA (Fig. 3 ). Several researchers have reported that applying CA enhances biomass and plant growth under metal stress in various plant species, including Juncus effusus ( Najeeb et al., 2009 ), Zea mays (Anwer et al., 2012 ), Iris halophile ( Han et al., 2018 ), and Brassica napus (Afshan et al., 2015 ). This elevation in biomass and plant growth may be attributed to improved plants’ nutrient uptake ( Najeeb et al., 2011 ) or phytochelatins (PCs) synthesis in plants (Muhammad et al., 2009 ). Additionally, applying VW promoted all the tested growth criteria, as shown in Fig. 3 . When considering the individual and combined applications of VW and BC, the influence on plant growth followed this order: K > VW > B2 > B1 > B1 + VW > B2 + VW > CA > CA + VW ( Fig. 3 ) . VW, a liquid bio-fertilizer with high water content, contains P, K, N, Zn, Ca, plant growth hormones, amino acids, and vitamins ( Sundararasu and Jeyasankar, 2014 ). Rathika et al. (2020) also found that applying VW improved the morphological characteristics of sorghum plants grown in Pb and Ni-polluted soil significantly, and similar results were reported by Naroila and Poonia (2011) for pearl millet. Impact of various amendments on pigments The data reveals that treatment with contaminated soil (K), without the addition of BC, substantially (p ≤ 0.05) diminished chlorophyll a, b, levels and total chlorophyll (Fig. 4 ). However, the addition of both tested BC led to a significant increase in chlorophyll content. The most substantial enhancements in chlorophyll content, specifically a 41% and 28% increase for chl. a, a 43% and 51% increase for chl. b, and a 32% and 29% increase in total chlorophyll were observed at a BC application rate of 1.5% for B1 and B2, respectively ( Fig. 4 ). Several studies conducted by Bashir et al. (2018), Mehmood et al. (2018) , and Helaoui et al. (2023) have reported that the application of BC derived from rice straw can elevate chlorophyll content and mitigate oxidative stress. Conversely, Belhaj et al. (2016) noted a decrease in chlorophyll a and b due to increased HM contents in soil contaminated by mining activities. Additionally, BC appeared to alleviate the stress caused by HMs on sorghum plants through mechanisms such as toxic metal adsorption onto the BC surface (Arabyarmohammadi et al., 2018 ), enhancement of soil physico-chemical properties, and enhanced nutrient uptake by sorghum ( Ok et al., 2015; Younis et al., 2016 ). Furthermore, Shahbaz et al. (2018) identified a substantial negative association between nickel concentration in the shoots and sunflower and maize plants as well as their physiological characteristics after incorporating BC. This result indicates that BC may accelerate the adsorption of HMs (through surface interactions), ultimately improving the photosynthetic capacity of the tested plants under HM stress. Conversely, compared to the K treatment, CA application alone elevated chl. a, b, and total chlorophyll by 86%, 82%, and 81%, respectively. In comparison, the application of VW alone increased these chlorophyll levels by 79%, 58%, and 68%, respectively, in plants subjected to HM stress. However, the most significant increases in chl. a, b, and total chlorophyll, at 105%, 80%, and 91%, respectively, were detected in plants grown (under the combined VW and CA application) ( Fig. 4 ). Afshan et al. ( 2015 ) illustrated that CA application significantly increased photosynthetic pigment levels in Brassica napus when grown in soil contaminated by Cr, compared to exposure to Cr alone. This increase in photosynthetic pigments may be attributed to enhanced antioxidant enzyme activity, which reduces the production of malondialdehyde (MDA) and electrolyte leakage. These findings are consistent with those of Mallhi et al. (2019), who found that photosynthetic pigments of castor plants grown in Pb-contaminated soil significantly enhanced after applying CA compared to treatments with Pb alone. The beneficial role of CA in the photosynthetic system of plants exposed to heavy metal stress has been described in numerous studies, including Shakoor et al. (2014) , Afshan et al. ( 2015 ), and Farid et al. (2017) . Applying CA enhances essential nutrients’ uptake and photosynthetic pigments’ formation ( Farid et al., 2017 ). Impact of various amendments on HMs concentration and uptake The addition of BC led to improved plant growth and alleviated the tested HM mobility and availability within the plant tissues. The substantial decline (p ≤ 0.05) in HM bioavailable concentration in soil induced the decreased translocation in the root and shoots of sorghum (grown in contaminated soil). For instance, when comparing the contents of Zn, Pb, Ni, Mn, Fe, Cu, and Cd in sorghum plants’ roots, the application of B1 and B2 reduced them from 3.10, 129.29, 170.61, 54.07, 7.96, 47.91, and 115.59 mg/kg to 2.66, 115.08, 149.86, 45.62, 7.19, 39.79, and 103.80 mg/kg, respectively for B1, and 2.66, 116.69, 142.39, 45.70, 7.10, 37.89, and 103.86 mg/kg, respectively for B2 (Table 6 ). Table 6 Concentration of Cd, Cu, Fe, Mn, Ni, Pb, and Zn (mg/Kg) on roots and shoots of sorghum plants at different treatments. Treatment Root Cd Cu Fe Mn Ni Pb Zn K 3.10 ± 0.54 b 129.29 ± 5.36 b 170.61 ± 6.17 b 54.07 ± 3.46 b 7.96 ± 0.76 ab 47.91 ± 2.54 b 115.59 ± 7.11 b VW 2.95 ± 0.28 b 126.86 ± 5.26 b 167.90 ± 7.10 b 56.21 ± 3.80 b 8.13 ± 1.09 ab 45.89 ± 2.78 bc 116.99 ± 6.13 b B1 2.66 ± 0.32 c 115.08 ± 5.89 c 149.86 ± 6.42 c 45.62 ± 2.67 c 7.19 ± 0.72 b 39.79 ± 2.60 d 103.80 ± 4.14 c B1 + VW 2.70 ± 0.23 c 115.33 ± 5.00 c 147.24 ± 6.14 c 44.74 ± 2.22 c 7.22 ± 0.86 b 39.00 ± 1.98 d 104.14 ± 4.28 c B2 2.66 ± 0.27 c 116.69 ± 7.13 c 142.39 ± 6.08 c 45.70 ± 3.28 c 7.10 ± 1.03 b 37.89 ± 3.90 d 103.86 ± 4.27 c B2 + VW 2.68 ± 0.21 c 116.18 ± 4.24 c 140.79 ± 5.13 c 46.66 ± 3.08 c 7.10 ± 0.87 b 40.89 ± 2.79 cd 102.07 ± 5.28 c CA 3.99 ± 0.35 a 142.92 ± 6.36 a 187.15 ± 8.07 a 64.32 ± 3.28 a 9.28 ± 1.10 a 53.77 ± 2.77 a 131.69 ± 6.36 a CA + VW 3.90 ± 0.24 a 144.77 ± 5.64 a 184.81 ± 6.13 a 65.36 ± 3.31 a 9.64 ± 1.24 a 56.91 ± 3.87 a 138.38 ± 4.76 a Shoot Cd Cu Fe Mn Ni Pb Zn K 1.36 ± 0.12 b 26.41 ± 2.82 ab 38.75 ± 3.25 bc 9.09 ± 0.53 b 3.70 ± 0.11 ab 8.83 ± 1.10 ab 20.78 ± 2.32 b VW 1.34 ± 0.05 c 24.54 ± 2.07 ab 39.87 ± 3.45 bc 8.33 ± 0.46 bc 3.78 ± 0.26 ab 8.69 ± 0.99 ab 20.44 ± 1.07 b B1 0.94 ± 0.09 c 22.95 ± 1.12 b 34.89 ± 2.19 cd 7.94 ± 0.22 bc 2.93 ± 0.12 c 7.28 ± 0.20 bc 17.98 ± 1.12 c B1 + VW 0.91 ± 0.07 c 23.06 ± 1.09 b 35.02 ± 3.22 cd 7.52 ± 0.57 c 2.99 ± 0.66 c 7.72 ± 1.19 bc 18.22 ± 2.08 c B2 0.96 ± 0.06 c 24.04 ± 1.20 b 32.85 ± 2.87 cd 8.10 ± 0.54 bc 2.85 ± 0.13 c 7.09 ± 1.06 bc 18.14 ± 2.10 c B2 + VW 0.93 ± 0.09 c 23.10 ± 2.10 b 33.82 ± 3.26 cd 8.20 ± 1.00 bc 2.92 ± 0.23 c 7.16 ± 0.64 bc 18.50 ± 1.11 c CA 1.55 ± 0.06 a 30.59 ± 1.79 a 45.87 ± 2.35 a 11.20 ± 2.15 a 4.20 ± 0.44 a 10.66 ± 1.13 a 25.44 ± 2.10 a CA + VW 1.49 ± 0.06 a 29.37 ± 1.65 a 43.51 ± 2.86ab 12.02 ± 1.13 a 4.11 ± 0.35 a 11.07 ± 1.22 a 26.09 ± 1.79 a All the data are presented as mean values of three independent experiments (n = 3). K represents un-amendment (control), VW (addition of vermiwash), B1 (biochar date palm), B1 + VW (biochar date palm + vermiwash), B2 (biochar Prosopis), B2 + VW (biochar Prosopis + vermiwash), CA (citric acid), and CA + VW (citric acid + vermiwash). All the means sharing common letter(s) are insignificantly different at P ≤ 0.05. Furthermore, it was observed that both B1 and B2 treatments significantly reduced the concentrations of Cd, Cu, Fe, Mn, Ni, Pb, and Zn in the shoots compared to the control soil. These reductions amounted to 30.88%, 13.10%, 9.96%, 12.65%, 20.81%, 17.55%, and 13.47%, respectively for B1, and 29.41%, 8.97%, 15.23%, 10.89%, 22.97%, 19.71%, and 12.70%, respectively for B2. The results demonstrate that applying BC immobilized HMs such as Cd, Cu, Fe, Mn, Ni, Pb, and Zn in the soil reduces their concentrations in the roots and shoots of sorghum plants. Numerous studies have previously reported similar findings, where BC amendments in heavy metal-contaminated soil resulted in the immobilization of these metals, leading to decreased uptake by plants ( Moon et al., 2013; Rehman et al., 2018; Yin et al., 2016; Gascó et al., 2019; Helaoui et al., 2023 ). In a study by Abbas et al. ( 2017 ), varying rice straw BC levels (0, 1.5, 3.0, and 5% w/w) were applied to Cd-contaminated soil. The outcomes indicated that BC treatments lowered Ni and Cd concentrations in wheat grains, roots, and shoots compared to controls. In addition, Cd concentration in wheat grains declined by 26%, 42%, and 57% following the application of 1.5%, 3.0%, and 5.0% BC, respectively, compared with the control. The reduction in HM concentrations in the roots of sorghum plants may be attributed to the selectivity of plants and metals’ immobilization within the soil, as evidenced by a decrease in DTPA-extracted soil HM levels ( Lu et al., 2012 ). Functional groups in the BC may have immobilized Cu and Pb concentrations ( Uchimiya et al., 2011 ). Immobilization of HMs through BC applications has been previously documented (Ahmad et al., 2016 ; Jones et al., 2016; Ok et al., 2015 ). Consequently, applying both tested BCs (B1 and B2) proved more effective in promoting plant growth and reducing metal concentrations in the roots and shoots of sorghum plants (Table 6 ) . However, the application of 1.5 mM CA to the plants increased the Cd, Cu, Fe, Mn, Ni, Pb, and Zn contents in the roots by 22.31%, 9.54%, 8.84%, 15.94%, 14.22%, 10.90%, and 12.23%, respectively, whereas in the shoots, increased by 12.26%, 13.66%, 15.52%, 18.84%, 11.90%, 17.17%, and 18.32%, respectively, compared to the plants grown in K treatment (Table 6 ). A comprehensive comprehension of the accumulation of HMs and the transportation behavior of plants is crucial in the process of selecting suitable agents for phytoremediation or enhancing plant stress tolerance. Organic acids have been widely recognized for their dual function in facilitating metal accumulation and acting as allies in the development of stress tolerance in plants involving metal tolerance ( Ehsan et al., 2014; Mahmud et al., 2017 ). Exogenous application of CA has been shown to enhance HM uptake in numerous plants ( Najeeb et al., 2011; Almaroai et al., 2013 ; Ehsan et al., 2014 ). CA in this study significantly increased the HM content in the shoots and roots of sorghum, confirming its potential for phytoremediation. The CA application also considerably elevated HM translocation from roots to shoots. Applying VW alone and combined with BC 1& 2 and CA has no significant effect on tested HM contents compared to treatments with the tested BC and chelating agents alone. The findings of this study provide evidence in favor of the hypothesis that the effectiveness of phytoextraction can be enhanced by augmenting the dry mass and, to a lesser degree, the metal accumulation in the upper parts of the plants (Table 6 ). Chelating agents are frequently employed to improve the availability of metals and promote their accumulation in plants. However, this increased metal uptake can lead to decreased plant biomass due to the toxicity associated with excessive metal absorption. Therefore, the simultaneous application of VW and CA represents an optimal approach to address this issue. Moreover, this combination can enhance a plant’s inherent antioxidative defense mechanisms, as suggested by Gao et al. (2010) . Conversely, the efficacy of the phytoremediation procedure, encompassing phytoextraction and phytostabilization, is contingent upon the quantity of biomass produced by the roots and shoots, as well as their capacity to absorb HMs. As a result, the quantification of metal absorption by the roots and shoots of sorghum plants was computed and graphically depicted in Fig. 5 . The findings of the study indicated a significant increase (p ≤ 0.05) in the uptake of heavy metals (HMs) by both the shoot and root when exposed to all tested treatments, as compared to the untreated soil potassium (K). The combined application of calcium (CA) and vermicompost (VW) resulted in the highest uptake of heavy metals (HM) in both the shoot and root. This observation can be attributed to the increased dry matter production associated with the application of VW, as compared to K. The concurrent utilization of CA and VW resulted in a significant enhancement in plants’ absorption of various heavy metals. Specifically, the uptake of Cd increased from 4.37 to 11.28 µg/pot, Cu increased from 163.48 to 408.66 µg/pot, Fe increased from 219.82 to 535.79 µg/pot, Mn increased from 66.32 to 179.42 µg/pot, Ni increased from 11.82 to 32.27 µg/pot, lead (Pb) increased from 59.58 to 159.53 µg/pot, and Zn increased from 143.19 to 385.95 µg/pot, respectively, in comparison to the control treatment using K. However, applying BC and VW improved the sorghum plant’s phytoextraction ability by enhancing plant growth. This study’s findings indicate that using BC in conjunction with nutrient supplements may be a viable approach for enhancing the phytoremediation of heavy metals (HMs) when employing sorghum as a bioenergy crop. The results obtained in this study were consistent with the findings reported by Houben et al. (2013), Fellet et al. (2014), and Houben et al. (2013) also observed the phytostabilization of cadmium (Cd) and zinc (Zn) by Brassica napus plants when BC was applied. A study conducted by Fellet et al. (2014) observed that the application of BC led to increased uptake of lead (Pb) by various plant species. This phenomenon was attributed to plant growth promotion, thereby indicating the potential for phytostabilization. The application of BC frequently leads to enhanced soil water-holding capacity, nutrient retention, microbial community composition, cation exchange capacity (CEC), and plant responses to diseases. The enhanced attributes of the BC ultimately lead to a higher crop output, indicating the BC’s potential as a phytoextractant ( Graber et al., 2010; Paz Ferreiro et al., 2014 ). Phytoextraction efficiency of metals The bioconcentration factors (BCF) and translocation factors (TF) for sorghum are presented in Figs. 6 & 7 . Baker (1981) has classified BCF into three distinct categories, which are as follows: Plants exhibiting a BCF below 1 are classified as excluders, while those falling within the range of 1 to 10 are classified as accumulators. Plants with BCF values exceeding 10 are referred to as hyperaccumulators. The HM BCF values for the sorghum plant were observed to follow a decreasing order, with Cd having the highest average value, followed by Cu and Fe, and then Mn, Ni, Pb, and Zn. The mean BCF value across all treatments was found to be below 1 ( Fig. 6 ). The study findings revealed that the transfer factor (TF) and bioconcentration factor (BCF) values for the various amendments examined were found to be less than one. This suggests that the process of phytoextraction is not viable for the sorghum plants in the present investigation despite their elevated biomass yield ( Steve and Zhao, 2003 ). Metal accumulation by the shoot and root of sorghum plants is depicted in Table 5 . The study results indicated that a considerable proportion of the metals was accumulated in the roots, as opposed to the shoots. The results presented in ( Figs. 6 & 7 ) indicate a decline in the translocation of HMs in the roots, as evidenced by the lower values of TF and BCF. This finding suggests that the phytostabilization process was not as effective in preventing the accumulation of HMs in the roots. Phytostabilization refers to the capacity of certain plant species to effectively accumulate a substantial amount of HMs in their root systems, thereby mitigating the risk of contamination in nearby or underground water sources (Vamerali et al., 2009). Fellet et al. (2014) and Karami et al. (2011) have documented that BC application has demonstrated suitability for the phytostabilization of certain HMs. Therefore, based on our findings, it can be suggested that the utilization of date palm and prosobis BC demonstrates effective potential for the immobilization of heavy metals in soil. However, the implementation of phytoextraction using sorghum plants in soil contaminated by mining activities is not considered a viable option. According to Goswami and Das (2015 ), soluble chemical elements have the ability to enter roots through two distinct pathways: the apoplastic pathway, which involves movement through the cell wall-free space, and the symplastic pathway, involving transportation across the plasma membrane of root cells followed by movement through the cytoplasm. Multiple studies have demonstrated that roots serve as a protective barrier against the translocation of HMs, thereby protecting the stems and other aboveground plant components from contamination and minimizing oxidative stress (Panwar et al., 2002; Liu et al., 2019). The observed variations in HM concentrations in different plant parts indicate the presence of distinct cellular mechanisms responsible for partitioning, translocating, and bioaccumulating these metals within plant systems (Sinha et al., 2007; Sharma and Dietz, 2009). Furthermore, HM translocation and uptake to the shoots are inherently connected to the speciation of soil organic matter, soil pH, SOM, and elements ( Kabata-Pendias, 2010 ). Multivariate Analysis A principal component analysis (PCA) It has been performed to evaluate the effect of different treatments in removing tested HMs (Fig. 8 ). Metals are correlated negatively with PCA components. In contrast, other metals correlate positively with PCA components. It was shown that some treatments are suitable for removing HMs; on the other hand, some are not suitable according to PCA analysis. B1, B2, B1 + VW, and B2 + VW are negatively correlated with PCA components for removing HMs. Meanwhile, VW, CA, and CA + VW are positively correlated with PCA for removing HMs. The most critical treatment used in this study to remove HMs is Vermiwash (VW), which is effectively necessary for removing Zn. The results indicated that Cu and Zn are correlated positively with PCA. In comparison, Cd and Ni correlated negatively with PCA components (Fig. 8 ). PCA 1 (91.82%) and PCA 2 (5.5%) explain the majority of the data, so there is no need for the other PCA components. Conclusion Phytoextraction is highly dependent on potentially toxic metals’ bioavailability in soil and plant characteristics. Remediation of soil contaminated with Cd, Cu, Fe, Mn, Ni, Pb, and Zn using both tested BC (data palm and Prosopysis), citric abid, and VW separately and together were investigated by performing a pot experiment for 90 days. The BC application significantly decreased the bioavailable tested HM contents, while the application of CA significantly enhanced the bioavailability of treated HMs in mine-contaminated soil. Compared to unamendment treatment (contaminated soil without any treatment), applying different treatments (BC, CA, and VW) separately and together led to significant enhancement of morphological traits and chlorophyll content of sorghum plants. Moreover, HM contents and uptake to sorghum shoot and root substantially (p ≤ 0.05) elevated with all tested treatments compared to the untreated soil (K). Furthermore, all tested amendments’ TF and BCF values are less than unity. This finding suggests that phytoextraction is not feasible for the sorghum plants in the current study despite higher biomass production. In contrast, VW application has been found to alleviate the detrimental impacts of HMs on both plant growth and photosynthetic efficiency. Additionally, when co-inoculated with contaminated soil, VW has been observed to enhance the capacity for phytoremediation. However, additional research is required to verify the results reproducibility and assess the potential for practical implementation in the field. Declarations Availability of data and materials: Not applicable. Ethics approval : Not applicable. Consent to participate: Not applicable. Consent to publish : Not applicable. Competing interests: The authors declare no competing interests. Author contribution statement: Hanan E. Osman and Ruwaydah Fadhlallah conceived and designed, and performed the experiments; Hanan E. Osman; Ruwaydah S. Fadhlallah and Mohamed H. 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Supplementary Files Suuplmenterydata10102024.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Major Revision 19 Mar, 2024 Reviewers agreed at journal 02 Jan, 2024 Reviewers invited by journal 09 Dec, 2023 Editor assigned by journal 24 Nov, 2023 First submitted to journal 19 Nov, 2023 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-3445202","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":258121068,"identity":"d1ebc38f-d9d3-41dc-883b-44e1772ea3bb","order_by":0,"name":"Hanan E. Osman","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzUlEQVRIiWNgGAWjYBACAygtB6HYSNBiTLqWxAaitZizNx/+dKPmTvqG4zkGDB/KDjPItx/Ar8Wy51iadM6xZ7kbzrwxYJxx7jCDwZkEAg67kWPGnMN2OHfDjRwDZt42oBYGQlruvzH+nPPvcLoBSMtfoBb5/geEbOExkM5tO5wA1sII1MJwg5AtZ9LSpHP7DhvOPPOs4GDPuXQegxuEbDl++PDnnG+H5fmOJ2988KPMWk6+n4AtSCCB4QCQ5CFaPVjLKBgFo2AUjAKsAADqJkk+dlqH6wAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0003-0612-842X","institution":"Umm Al-Qura University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Hanan","middleName":"E.","lastName":"Osman","suffix":""},{"id":258121069,"identity":"739c9882-0f3c-4622-b6ed-2b5626ce761e","order_by":1,"name":"Ruwaydah S. Fadhlallah","email":"","orcid":"","institution":"Umm Al-Qura University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ruwaydah","middleName":"S.","lastName":"Fadhlallah","suffix":""},{"id":258121070,"identity":"ff133f9d-d269-48e8-bff1-f13fb6da2090","order_by":2,"name":"Mohamed H.E. El-Morsy","email":"","orcid":"","institution":"Umm Al-Qura University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"H.E.","lastName":"El-Morsy","suffix":""}],"badges":[],"createdAt":"2023-10-14 09:51:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3445202/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3445202/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":48066083,"identity":"1f9008d8-e51a-4cfa-9acf-2f0a4d82bf0f","added_by":"auto","created_at":"2023-12-12 15:08:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":773113,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePreparation of vermiwash. The bottom layer contains stone pieces in coarse grain. The next layer has sand, which helps drain water to maintain moderate moisture. In the third layer, earthworms are inoculated with organic soil, in the fourth layer.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/3a6b0952cd1e9c0c8d532f26.png"},{"id":48066082,"identity":"1ae01096-b507-4953-8d40-688ad237d587","added_by":"auto","created_at":"2023-12-12 15:08:58","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":977383,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSite location of the studied area.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/ea76f3f023463e89e368582e.png"},{"id":48066085,"identity":"7feeb915-cba2-4422-b781-6e2b6e645107","added_by":"auto","created_at":"2023-12-12 15:08:58","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":329014,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRoot and shoot length, and biomass fresh and dry weights (g/pot) of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003esorghum bicolor\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003egrown in mine-contaminated soil. K represents un-amendment (control), VW (addition of vermiwash), B1 (biochar date palm), B1+ VW (biochar date palm+ vermiwash), B2 (biochar Prosopis), B2+VW (biochar Prosopis +vermiwash), CA (citric acid), and CA+VW (citric acid + vermiwash). All the means sharing common letter(s) are insignificantly different at P ≤ 0.05 level.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/6bc190e4e9d2f223150610f2.png"},{"id":48066081,"identity":"89a41f63-d87f-475d-a9ae-3b45df71ae05","added_by":"auto","created_at":"2023-12-12 15:08:58","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":240797,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChlorophyll a, b, and total chlorophyll contents of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eSorghum bicolor\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e grown in mine contaminated soil. K represents un-amendment (control), VW (addition of vermiwash), B1 (biochar date palm), B1+ VW (biochar date palm+ vermiwash), B2 (biochar Prosopis), B2+VW (biochar Prosopis +vermiwash), CA (citric acid), and CA+VW (citric acid + vermiwash). All the means sharing common letter(s) are insignificantly different at P ≤ 0.05.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/2b433db7e537a27d6a80215e.png"},{"id":48066078,"identity":"ee0cd6bf-26aa-4451-ae97-a6ebf89d566a","added_by":"auto","created_at":"2023-12-12 15:08:57","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":946603,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eUptake (mg/plant) of Cd, Cu, Fe, Mn, Ni, Pb, and Zn on sorghum plants at different treatments. All the data are presented as mean values of three independent experiments (n = 3). K represents un-amendment (control), VW (vermiwash), B1 (biochar date palm), B1+ VW (biochar date palm+ vermiwash), B2 (biochar Prosopis), B2+VW (biochar Prosopis +vermiwash), CA (citric acid), and CA+VW (citric acid + vermiwash). All the means sharing common letter(s) are insignificantly different at P ≤ 0.05.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/c467a852b22009582aa92c87.png"},{"id":48066077,"identity":"806d88ff-2be7-41d2-8980-bf2038e83e89","added_by":"auto","created_at":"2023-12-12 15:08:57","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":16766,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMean of bioconcentration value of Cd, Cu, Fe, Mn, Ni, Pb and Zn of sorghum plant under different treatments. All the data are presented as mean values (n = 3), and error bars represent SD. K represents un-amendment (control), VW (addition of vermiwash), B1 (biochar date palm), B1+ VW (biochar date palm+ vermiwash), B2 (biochar Prosopis), B2+VW (biochar Prosopis +vermiwash), CA (citric acid), and CA+VW (citric acid + vermiwash). All the means sharing common letter(s) are insignificantly different at P ≤ 0.05.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/04f80926d894f0106c453d52.png"},{"id":48067982,"identity":"54414a41-47c3-440a-9292-ea990177bed7","added_by":"auto","created_at":"2023-12-12 15:16:58","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":13582,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTranslocation Factor (TF) of Cd, Cu, Fe, Mn, Ni, Pb, and Zn of sorghum plant under different treatments. All the data are presented as mean values (n = 3), and error bars represent SD.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/b2cdcc98b797e19e188378d1.png"},{"id":48066080,"identity":"4398b183-7f91-4f8f-a527-f9c80cf30c46","added_by":"auto","created_at":"2023-12-12 15:08:58","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":244202,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePCA analysis of heavy metal removal under different treatments.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/f26f431a4a1fbb55196cfbf4.png"},{"id":48069312,"identity":"33ca4201-9bcd-4d2c-85bd-de55f5547440","added_by":"auto","created_at":"2023-12-12 15:26:44","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6150618,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/6dfb2f02-8b94-44ed-bf28-2e4065c0c420.pdf"},{"id":48066084,"identity":"420b832f-843d-4687-9f82-577eccb07a67","added_by":"auto","created_at":"2023-12-12 15:08:58","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":6140496,"visible":true,"origin":"","legend":"","description":"","filename":"Suuplmenterydata10102024.docx","url":"https://assets-eu.researchsquare.com/files/rs-3445202/v1/cf6074ccd25e0c6fdee18e47.docx"}],"financialInterests":"","formattedTitle":"Synergistic phytoremediation effect by Sorghum bicolor, citric acid, biochar, and Vermiwash amendment for the remediation of a mine-contaminated soil","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAs an integral component of the Earth\u0026rsquo;s ecosystem, it serves a crucial function in sustaining the quality of life for humans as well as the survival of various macro and microorganisms. Unfortunately, numerous detrimental substances are being released into the environment (as liquid and solid waste) by mining and industrial operations, leading to significant pollution in the surrounding areas (Baldauf et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). In addition, m activities have led to the alteration of the natural landscape, the contamination of water and soil resources, and soil compaction through the discharge of numerous hazardous substances (\u003cb\u003eRodriguez-Franco and Page-Dumroes 2021\u003c/b\u003e). Among these harmful substances, HMs are frequently found near waste disposal sites, causing substantial water as well as soil contamination (\u003cb\u003eGranero and Domingo 2002; Nadal et al. 2005; Ferreira et al., 2021\u003c/b\u003e). Such contaminated soil is unsuitable for agricultural practices, as it can accumulate HMs in edible crops, rendering them unfit for consumption (\u003cb\u003eChen et al. 2015; Jia et al. 2018\u003c/b\u003e). Therefore, restoring these polluted sites using economically viable techniques is imperative before they can be utilized for agricultural purposes, ensuring their suitability for productive farming activities (\u003cb\u003eOladoye et al., 2022; Quronfulah et al., 2023)\u003c/b\u003e.\u003c/p\u003e \u003cp\u003eTraditional soil remediation methods involve destructive practices like excavation, landfilling, and acid washing. However, there is a growing interest in developing and utilizing cost-effective, in-situ, and environmentally friendly technologies for soil decontamination and metal recovery. Phytoremediation methods, which involve the utilization of particular plant species for the purposes of immobilizing contaminants (phytostabilization), extracting them from the environment (phytoextraction), or converting them into less detrimental forms, have garnered significant attention (\u003cb\u003ePrasad, 2003; Vangronsveld et al., 2009;\u003c/b\u003e Alegbeleye et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; \u003cb\u003eOladoye et al., 2022\u003c/b\u003e). In order to render contaminated lands suitable for agricultural activities, they must be restored using economically feasible techniques before being used for agricultural production. Stabilization of HMs-contaminated soil in situ has been deemed viable, cost-effective, and simple to implement. Several additives have been used to stabilize in situ, including chelating agents, organic adsorbents, compost, biosolids, and activated carbon (\u003cb\u003eRafique et al., 2020\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eOver the past few years, various phytoremediation approaches have been employed, with processes involving organic amendments gaining increased attention (\u003cb\u003eWiszniewska et al., 2016\u003c/b\u003e). These approaches encompass the use of organic acids (decreased molecular weight) (\u003cb\u003eDuarte et al., 2011; Chen et al., 2020; Helaoui et al., 2023; Wang et al., 2023\u003c/b\u003e), the introduction of plant growth-promoting substances (e.g., VW through bioaugmentation) (\u003cb\u003eParedes-Paliz et al., 2018\u003c/b\u003e), and in recent times, the implementation of BC (\u003cb\u003eGhosh and Maiti, 2021; Wang et al., 2023\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eBiochar (BC), a product of biomass pyrolysis, is used in various environmental applications because it is a cost-effective, carbonaceous material with marked efficacy and versatility and a large surface area with numerous active sites, particularly as a soil amendment for HM remediation (\u003cb\u003eHe et al., 2019; Ghosh and Maiti, 2021\u003c/b\u003e). The selection of raw materials utilized in the manufacturing of biochar and the specific conditions applied during the pyrolysis process are crucial factors that have a significant influence on the characteristics of biochar-metal binding and its impacts on soil (\u003cb\u003eQi et al., 2017; Fan et al., 2020; Harindintwali et al., 2020\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eAnother viable approach to promote the solubility of potential soil\u0026rsquo;s toxic metals is adding chelating agents. Chelates like organic acids (of low molecular weight) boost plants\u0026rsquo; uptake of potentially toxic metals (Anning and Akoto, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; \u003cb\u003eYin et al., 2015\u003c/b\u003e). CA has been utilized as an amendment to enhance the elimination of different metals owing to its environmentally sustainable characteristics, facile biodegradability, and non-toxic characteristics (Anwer et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Al Mahmud et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Numerous studies have proven that CA significantly contributes to phytoremediation enhancement in multiple metals by improving metal solubility and mobilization. In addition, CA contributes to mitigating metal-induced oxidative stress by enhancing the activity of enzymes in antioxidative defense systems (Al Mahmud et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe primary objectives of all these organic amendments are twofold: i) to enhance elements\u0026rsquo; phytostabilization or photo-availability for extraction, and ii) to promote plant resistance as well as tolerance to the uptake of potentially toxic metals (\u003cb\u003eDuarte et al., 2011; Ghosh and Maiti, 2021\u003c/b\u003e). These objectives aim to improve phytoremediation performance regarding element extraction, plant health, and subsequent biomass production.\u003c/p\u003e \u003cp\u003eHowever, there is limited research on using VW and its effects on plants cultivated in contaminated soils. VW, an extract from vermicomposting, is a brown-colored liquid rich in plant growth-promoting hormones, calcium, zinc, potassium, amino acids, vitamins, and nitrogen (Aghamohammadi et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Numerous studies have been conducted to investigate the potential of VW as a bio-fertilizer and pesticide for augmenting crops\u0026rsquo; productivity and growth.\u003c/p\u003e \u003cp\u003eThe study is designed to achieve the following goals: 1) Investigate the combined impact of CA, BC, and VW on the phytoremediation of mine-contaminated soil. 2) Evaluate how different amendments affect sorghum plants\u0026rsquo; growth, biomass, and chlorophyll content. 3) Examine the tested HM accumulation in sorghum plants cultivated in mine-contaminated soil.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eVermiwash preparation and analysis\u003c/h2\u003e \u003cp\u003eVermiwash (VW) was produced through the secretion of \u003cem\u003eEudrillus euginiae\u003c/em\u003e earthworms, extracted as the seepage drained from the worm bed, and then preserved in earthen pots (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), following the established technique described by \u003cb\u003eIsmail (1997)\u003c/b\u003e. This collected VW was utilized in its undiluted form, i.e., 100% concentration, the characteristics of the VW (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cb\u003eChemical properties of vermiwash (VW) fertilizer used\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCd\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003ePb\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003eN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003eTOC\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c10\" namest=\"c3\"\u003e \u003cp\u003e\u003cb\u003emg/L\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e \u003cp\u003e\u003cb\u003e%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003emg/l\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e7.65\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e1.78\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e4.38\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003end\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.03\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e0.25\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e1.50\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.97\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003end\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003end\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e1.71\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e0.522\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e521.88\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe electrical conductivity (EC) as well as chemical reactions (pH) were examined using the methodology outlined by \u003cb\u003eGarg et al. (2006)\u003c/b\u003e. Additionally, total organic carbon (TOC) content was assessed following the procedure established by \u003cb\u003eNelson and Sommers (1982)\u003c/b\u003e. Total nitrogen (N) was quantified using the method illustrated by \u003cb\u003eBremmer and Mulvaney (1982)\u003c/b\u003e. Finally, total P, K, Na, and Ca were determined following the procedure described by Bansal and Kapoor (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2000\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eProduction and characterization of tested BC\u003c/h2\u003e \u003cp\u003eDate palm (\u003cem\u003ePhoenix dactylifera\u003c/em\u003e) and Prosopis (\u003cem\u003eProsopis juliflora\u003c/em\u003e) were picked, dried, chopped, and then placed (in a container made of airtight stainless-steel) with a 7 cm diameter and a height of 22 cm. Subsequently, they underwent pyrolysis in an electrical muffle furnace at 450\u0026deg;C for 4 hours (\u003cb\u003eAl-Wabel et al. 2013; Usman et al. 2015\u003c/b\u003e). Both biochar samples were subsequently pulverized and filtered using a 1.0-mm sieve (\u003cb\u003eSupplementary Fig.\u0026nbsp;1\u003c/b\u003e). The physiochemical properties of these biochar samples are outlined in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The EC and pH were determined using an aqueous extract of the BC at a 1:10 ratio. The analysis of dried BC samples was done in terms of their C, H, N, and S content using a CHNS analyzer (Vario EL III, Germany). X-ray diffraction (MAXima X XRD-7000-Shimadzu-Japan) was employed to determine any crystal structures formed in the materials (\u003cb\u003eSupplementary Figs.\u0026nbsp;2\u0026amp;3\u003c/b\u003e). Furthermore, examination of BC surface morphology was done via scanning electron microscopy (SEM) utilizing (JSM-6460 LV-Scanning Microscope) (\u003cb\u003eSupplementary Figs.\u0026nbsp;4\u0026amp;5\u003c/b\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhysiochemical properties of used biochar (n\u0026thinsp;=\u0026thinsp;3)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ePhysiochemical properties\u003c/span\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eEC\u003c/p\u003e \u003cp\u003e(mS/cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c8\" namest=\"c4\"\u003e \u003cp\u003eChemical composition (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eC\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eH\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eN\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003eS\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003eC/N\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003eratio\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDate palm\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e56.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e2.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e93.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eProsopis\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e72.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e120.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eGreenhouse experiment and soil and plant sampling\u003c/h2\u003e \u003cp\u003eThis study collected soil from the surface layer at the Mahad AD\u0026rsquo;Dahab gold deposit site. This mining location is 380 km northeast of Jeddah, precisely at coordinates 23\u0026deg;30\u0026rsquo;30\u0026rdquo; N latitude and 40\u0026deg;51\u0026rsquo;30\u0026rdquo; E longitude, in the late Proterozoic Arabian Shield\u0026rsquo;s west-central region (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eA controlled pot experiment was conducted at (Umm Al-Qura University, Saudi Arabia) at coordinates 21\u0026deg;18\u0026rsquo;56.3\u0026rdquo; N latitude and 39\u0026deg;56\u0026rsquo;47.8\u0026rdquo; E longitude from November 2020 to January 2021. The greenhouse setting at temperatures ranging from 20\u0026deg;C to 28\u0026deg;C and a humidity level of 50\u0026thinsp;\u0026plusmn;\u0026thinsp;4%. A total of 10 sorghum seeds were sown in individual pots containing 4 Kg of mine-contaminated soil. The present study adopted a completely randomized design (CRD), with each treatment replicated six times, as depicted in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. After 60 and 80 days from sowing, VW was applied as 20 ml/kg soil.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTreatments in this study.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDescription\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eK\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003econtrol\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eVemiwash\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ebiochar date palm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB1\u0026thinsp;+\u0026thinsp;VW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ebiochar date palm\u0026thinsp;+\u0026thinsp;vermiwash\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ebiochar Prosopis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eB2\u0026thinsp;+\u0026thinsp;VW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ebiochar Prosopis\u0026thinsp;+\u0026thinsp;vermiwash\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecitric acid\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCA\u0026thinsp;+\u0026thinsp;VW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecitric acid\u0026thinsp;+\u0026thinsp;vermiwash\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eIn contrast, CA was utilized as a chelating agent at a 1.5 mmol/kg soil rate after 80 days from plantation. Additionally, BC was utilized at a (1.5% / kg soil) rate before cultivation. The moisture levels were adjusted to 60% of the soil\u0026rsquo;s field capacity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eSoil Sampling and Analysis\u003c/h2\u003e \u003cp\u003ePrior to initiating study procedures, soil samples were obtained from each individual pot. The soil samples underwent the process of air-drying and were subsequently passed through a mesh (with a 2 mm pore size). The main physicochemical characteristics of soil collected from Mahad Adh Dahab were assessed by methods described in \u003cb\u003eBurt (2004)\u003c/b\u003e and are outlined in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Soil pH and EC were 7.66 and 1.68 ds/m, respectively. Additionally, the soil exhibited a sandy loam texture.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSoil characteristic, total, and DTPA- extracted Cd, Cu, Fe, Mn, Ni, Pb, and Zn contents in soil collected from Mahad Adh Dahab used in the pot experiment. (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD, n\u0026thinsp;=\u0026thinsp;3).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"12\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCd\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePb\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eEC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eOM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003eTexture\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"8\" nameend=\"c9\" namest=\"c2\"\u003e \u003cp\u003emg/Kg\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eDs/m\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1570.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e790.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e985.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e23.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e491.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1150.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e7.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e1.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e4.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eSandy loam\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSD\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e23.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e57.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e27.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e10.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e25.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDTPA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e169.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e56.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e78.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e43.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e116.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSD\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e7.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e4.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAcid mixture was utilized to digest the soil samples (HClO\u003csub\u003e4\u003c/sub\u003e: H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e: HNO\u003csub\u003e3\u003c/sub\u003e; 1:1:5, v/v/v) to evaluate the total HM contents (Zn, Pb, Cd, Ni, Cu, Mn, and Fe) (\u003cb\u003eUSEPA, 1992\u003c/b\u003e), and their concentrations were identified utilizing an inductively coupled plasma optical emission spectrophotometer (ICPOES; PerkinElmer Optima 4300 DV, USA). Diethylene triamine pentaacetate (DTPA) was used to extract and assess the available tested HM contents in the soil before and after cultivation in each treatment, according to \u003cb\u003eLindsay and Norvell (1978)\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003ePlant Sampling and Analysis\u003c/h2\u003e \u003cp\u003eThe samples were collected 90 days after the plantation. From each treatment, a suitable number of plant species were randomly selected and combined into three replicate groups for assessing growth criteria. The plants were divided into their root and shoots, and the lengths of both roots and shoots were recorded. Furthermore, the dry and fresh weights (g/ pot) were determined through the weight of different plant parts before and after oven-drying at 65\u0026deg;C until a constant weight was achieved.\u003c/p\u003e \u003cp\u003eThe photosynthetic pigments in fresh leaves were extracted with 85% acetone, and their concentrations were measured with a spectrophotometer at 662 nm, 644 nm, and 440.5 nm following the procedure Arnon (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1956\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. Moreover, the soil analysis procedures assessed seven tested HM contents in plant samples.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eData Analysis\u003c/h2\u003e \u003cp\u003eThe bioaccumulation factor (BCF) is a vital instrument for the prediction of HM absorption by plants from the soil. In contrast, HM translocation from roots to shoots is assessed using the translocation factor (TF) in order to determine the efficacy of different treatments in extracting metals from the soil and transporting them to the leaves (\u003cb\u003eGhosh and Singh, 2005\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eBCF\u0026thinsp;=\u0026thinsp;C\u003csub\u003eplant\u003c/sub\u003e / C\u003csub\u003esoil\u003c/sub\u003e (1)\u003c/p\u003e \u003cp\u003eTF\u0026thinsp;=\u0026thinsp;C\u003csub\u003eshoot\u003c/sub\u003e / C\u003csub\u003eroot\u003c/sub\u003e (2)\u003c/p\u003e \u003cp\u003eWhere C\u003csub\u003esoil\u003c/sub\u003e and C\u003csub\u003eplant\u003c/sub\u003e are the mean of shoot and root and HM concentration (mg/kg) in the soil, respectively.\u003c/p\u003e \u003cp\u003eHM uptake (\u0026micro;g/pot) was also conducted, as described by \u003cb\u003eUtmazian et al. (2007)\u003c/b\u003e.\u003c/p\u003e \u003cp\u003eTU (\u0026micro;g/pot)\u0026thinsp;=\u0026thinsp;HM concentration in roots or shoots dw X d wt of roots or shoots g/pot (3).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eThe data underwent analysis through a one-way ANOVA using the 22nd version of the IBM-SPSS software. Pearson\u0026rsquo;s correlation analysis was done to determine the associations between variables and assess their statistical significance with a confidence interval of 95%, and significance was determined at a threshold of p\u0026thinsp;\u0026le;\u0026thinsp;0.05. Similarly, multivariate statistics analysis is applied to categorize and evaluate the most suitable method for the removal of HMs. The data can be condensed, unified, and categorized using multivariate approaches to extract meaningful information. The technique can also be used to explain temporal and spatial variations brought on by relationships between seasonality and natural and human influences. The principal component analysis is used to analyze the data collected. The most popular method for converting the original variables into new, uncorrelated variables (axes) or principal components is principal component analysis (PCA). PCA explains data variations clearly (\u003cb\u003ePop et al. 2009\u003c/b\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eImpact of different treatments of DTPA \u0026ndash; heavy metal contents\u003c/h2\u003e \u003cp\u003eHM toxicity in soil is primarily impacted by the availability of metal forms accessible to plants rather than the overall concentration of metals in the soil (\u003cb\u003eYin et al., 2016\u003c/b\u003e). Zn, Pb, Ni, Cd, Mn, Fe, and Cu concentrations before and after cultivating sorghum plants in Mahad AD\u0026rsquo;Dahab under various treatment conditions are reported in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The findings indicate that the use of both tested BC in the soil led to a decrease in DTPA-extractable Zn, Pb, Ni, Cd, Mn, Fe, and Cu (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e), which is consistent with the outcomes demonstrated by \u003cb\u003eYousaf et al. (2016)\u003c/b\u003e for Cd and \u003cb\u003eRehman et al. (2016)\u003c/b\u003e for Ni.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDTPA- extracted Cd, Cu, Fe, Mn, Ni, Pb, and Zn contents before and after cultivation for 90 days grown in Mahad AD\u0026rsquo;Dahab used in the pot experiment. (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD, n\u0026thinsp;=\u0026thinsp;3)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCd\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePb\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003e\u003cspan type=\"BoldUnderline\" class=\"BoldUnderline\" name=\"Emphasis\"\u003eBefore cultivation\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e169.52\u0026thinsp;\u0026plusmn;\u0026thinsp;7.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e56.80\u0026thinsp;\u0026plusmn;\u0026thinsp;4.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e78.86\u0026thinsp;\u0026plusmn;\u0026thinsp;3.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e43.88\u0026thinsp;\u0026plusmn;\u0026thinsp;4.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e116.03\u0026thinsp;\u0026plusmn;\u0026thinsp;6.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003e\u003cspan type=\"BoldUnderline\" class=\"BoldUnderline\" name=\"Emphasis\"\u003eAfter Cultivation\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e155.57\u0026thinsp;\u0026plusmn;\u0026thinsp;5.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e49.70\u0026thinsp;\u0026plusmn;\u0026thinsp;4.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e69.80\u0026thinsp;\u0026plusmn;\u0026thinsp;6.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e39.12\u0026thinsp;\u0026plusmn;\u0026thinsp;3.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e103.76\u0026thinsp;\u0026plusmn;\u0026thinsp;7.15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eVW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e149.38\u0026thinsp;\u0026plusmn;\u0026thinsp;6.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e51.94\u0026thinsp;\u0026plusmn;\u0026thinsp;4.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e67.68\u0026thinsp;\u0026plusmn;\u0026thinsp;5.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.71\u0026thinsp;\u0026plusmn;\u0026thinsp;1.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e41.75\u0026thinsp;\u0026plusmn;\u0026thinsp;3.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e104.24\u0026thinsp;\u0026plusmn;\u0026thinsp;6.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e137.44\u0026thinsp;\u0026plusmn;\u0026thinsp;6.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e42.46\u0026thinsp;\u0026plusmn;\u0026thinsp;3.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e60.73\u0026thinsp;\u0026plusmn;\u0026thinsp;4.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e35.25\u0026thinsp;\u0026plusmn;\u0026thinsp;3.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e94.72\u0026thinsp;\u0026plusmn;\u0026thinsp;4.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB1\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e140.11\u0026thinsp;\u0026plusmn;\u0026thinsp;6.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e40.88\u0026thinsp;\u0026plusmn;\u0026thinsp;2.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e61.13\u0026thinsp;\u0026plusmn;\u0026thinsp;5.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e37.60\u0026thinsp;\u0026plusmn;\u0026thinsp;3.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e91.39\u0026thinsp;\u0026plusmn;\u0026thinsp;5.25\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e128.10\u0026thinsp;\u0026plusmn;\u0026thinsp;4.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e41.23\u0026thinsp;\u0026plusmn;\u0026thinsp;3.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e59.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e38.47\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e93.65\u0026thinsp;\u0026plusmn;\u0026thinsp;5.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB2\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e127.19\u0026thinsp;\u0026plusmn;\u0026thinsp;7.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e39.13\u0026thinsp;\u0026plusmn;\u0026thinsp;3.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e57.22\u0026thinsp;\u0026plusmn;\u0026thinsp;4.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40.01\u0026thinsp;\u0026plusmn;\u0026thinsp;3.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e96.27\u0026thinsp;\u0026plusmn;\u0026thinsp;3.61\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e188.04\u0026thinsp;\u0026plusmn;\u0026thinsp;4.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e65.97\u0026thinsp;\u0026plusmn;\u0026thinsp;2.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e92.16\u0026thinsp;\u0026plusmn;\u0026thinsp;5.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e49.32\u0026thinsp;\u0026plusmn;\u0026thinsp;3.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e130.77\u0026thinsp;\u0026plusmn;\u0026thinsp;5.49\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCA\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e183.41\u0026thinsp;\u0026plusmn;\u0026thinsp;7.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e64.75\u0026thinsp;\u0026plusmn;\u0026thinsp;3.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e90.41\u0026thinsp;\u0026plusmn;\u0026thinsp;5.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.76\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e47.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e127.81\u0026thinsp;\u0026plusmn;\u0026thinsp;5.59\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eSimilarly, the outcomes show that the bioavailability of Cd, Cu, Fe, Mn, Ni, Pb, and Zn decreased by 18.92%, 25.00%, 25.25%, 22.99%, 26.05%, 19.67%, and 18.37%, respectively, when date palm (BC1) biochar was applied. In contrast, Prosopis biochar (BC2) reduced 35.00%, 24.43%, 27.41%, 24.77%, 35.46%, 12.33%, and 19.29%, respectively (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). These findings confirm that date palm and Prosopis BC have an increased adsorption capacity and the potential to create binding sites for the tested HMs. This finding aligns with previous studies (\u003cb\u003eZheng et al., 2016;\u003c/b\u003e Bashir et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; \u003cb\u003eRafique et al., 2021; Ram\u0026iacute;rez et al., 2022; Wang et., 2022;\u003c/b\u003e Alazzaz et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; \u003cb\u003eLi et al., 2023)\u003c/b\u003e,\u003c/p\u003e \u003cp\u003eMoreover, the strong affinity of HMs to BC may be another reason for reduced metal availability. BC\u0026rsquo;s larger surface area and numerous contact sites significantly reduce DTPA-extractable Cu, Cd, Fe, Ni, Mn, Zn, and Pb in the soil (\u003cb\u003eLu et al., 2017; Naeem et al., 2020\u003c/b\u003e). Comparable results have been reported in prior research, where BC from various sources effectively reduced Cd, Cu, Fe, Mn, Ni, Pb, and Zn concentrations in mine-contaminated soil (\u003cb\u003eXu et al., 2016; Irfan et al., 2021;\u003c/b\u003e Alazzaz et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFurthermore, HM bioavailability in soil is crucial for successful phytoremediation (\u003cb\u003eGreman et al., 2001\u003c/b\u003e). In this study, the application of 1.5 mol/Kg soil of CA elevated the bioavailability of Zn, Pb, Ni, Mn, Fe, Cu, and Cd by 11.27%, 10.99%, 15.68%, 14.43%, 13.90%, 9.85%, and 16.96% respectively (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). The same results were recorded by \u003cb\u003eDiarra et al. (2021)\u003c/b\u003e, who found that applying CA released the most elevated Cu and Ni fractions in the soil, accounting for 75.5% and 79.5%, respectively. This finding is compatible with those of \u003cb\u003eWuana et al. (2010)\u003c/b\u003e, who revealed a significant decline in Cu and Ni levels through batch soil washing with CA. Furthermore, fractionation patterns revealed that CA primarily targets HMs related to the reducible as well as exchangeable fractions. A smaller proportion of HMs bound to the soil organic matter are also targeted by CA (\u003cb\u003eWuana et al., 2010\u003c/b\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eImpact of various amendments on morphological traits\u003c/h2\u003e \u003cp\u003ePlant morphological traits, such as plant height, root length, and plant biomass (both fresh and dry weights), were significantly influenced by various treatments when compared to the untreated treatment (K)(Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Root and shoot lengths varied between 9.33\u0026ndash;12.83 cm and 76.33-88.00 cm, respectively. The sorghum plants cultivated in untreated soil (K) exhibited the shortest root and shoot lengths, while the application of a combination of CA\u0026thinsp;+\u0026thinsp;VW to the soil led to the longest shoot and root lengths.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eCompared to control (K), the implementation of both tested BCs (B1\u0026amp;B2) significantly enhanced sorghum morphological characteristics. The enhancement in plant height, root length, and fresh and dry biomass was 20.61%, 59.72%, 83.06%, and 54.92% for the B1 treatment and 60.98%, 22.87%, 75.28%, and 61.59% for the B2 treatment.\u003c/p\u003e \u003cp\u003eAbbas et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) demonstrated that the application of BC substantially impacted the root length, plant height, and biomass of wheat plants when exposed to various Cd treatments compared to untreated conditions. BC soil amendment has been shown to promote the growth of numerous species of plants exposed to metal stress (\u003cb\u003eRizwan et al., 2016; Rehman et al., 2016; Younis et al., 2016;\u003c/b\u003e Almaroai and Eissa \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; \u003cb\u003eHelaoui et al., 2023\u003c/b\u003e). In this study, the elevation in these parameters by adding BC may be attributed to increased mineral nutrient availability. Previous research has frequently reported that BC improves plant growth because of improved biological characteristics, increased nutrient content, elevated cation exchange capacity, and pH adjustments (\u003cb\u003ePaz-Ferreiro et al., 2014; Širi\u0026rsquo;c et al., 2022\u003c/b\u003e). Additionally, \u003cb\u003eIrfan et al. (2021)\u003c/b\u003e found that applying wheat straw BC in polluted soil significantly enhanced shoot and root lengths and plant biomass in maize plants compared to those grown in contaminated soil without BC. Similar findings were reported by \u003cb\u003eLu et al. (2014)\u003c/b\u003e for edible amaranth, \u003cb\u003ePark et al. (2011)\u003c/b\u003e for canola plants, and Ali et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) for Indian mustard.\u003c/p\u003e \u003cp\u003eIn contrast, the application of CA resulted in a significant enhancement of shoot and root lengths, as well as overall plant biomass, in contaminated soil. Fresh and dry weights increased with CA application from 23.5 g and 3.15 g /pot in the K to 48.86 g and 5.85 g/pot. Additionally, sorghum root elongation, plant height, and biomass increased when VW was applied with BCand CA (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSeveral researchers have reported that applying CA enhances biomass and plant growth under metal stress in various plant species, including \u003cem\u003eJuncus effusus\u003c/em\u003e (\u003cb\u003eNajeeb et al., 2009\u003c/b\u003e), \u003cem\u003eZea mays\u003c/em\u003e (Anwer et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), \u003cem\u003eIris halophile\u003c/em\u003e (\u003cb\u003eHan et al., 2018\u003c/b\u003e), and \u003cem\u003eBrassica napus\u003c/em\u003e (Afshan et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). This elevation in biomass and plant growth may be attributed to improved plants\u0026rsquo; nutrient uptake (\u003cb\u003eNajeeb et al., 2011\u003c/b\u003e) or phytochelatins (PCs) synthesis in plants \u003cb\u003e(Muhammad et al., 2009\u003c/b\u003e). Additionally, applying VW promoted all the tested growth criteria, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eWhen considering the individual and combined applications of VW and BC, the influence on plant growth followed this order: K\u0026thinsp;\u0026gt;\u0026thinsp;VW\u0026thinsp;\u0026gt;\u0026thinsp;B2\u0026thinsp;\u0026gt;\u0026thinsp;B1\u0026thinsp;\u0026gt;\u0026thinsp;B1\u0026thinsp;+\u0026thinsp;VW\u0026thinsp;\u0026gt;\u0026thinsp;B2\u0026thinsp;+\u0026thinsp;VW\u0026thinsp;\u0026gt;\u0026thinsp;CA\u0026thinsp;\u0026gt;\u0026thinsp;CA\u0026thinsp;+\u0026thinsp;VW \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e. VW, a liquid bio-fertilizer with high water content, contains P, K, N, Zn, Ca, plant growth hormones, amino acids, and vitamins (\u003cb\u003eSundararasu and Jeyasankar, 2014\u003c/b\u003e). \u003cb\u003eRathika et al. (2020)\u003c/b\u003e also found that applying VW improved the morphological characteristics of sorghum plants grown in Pb and Ni-polluted soil significantly, and similar results were reported by \u003cb\u003eNaroila and Poonia (2011)\u003c/b\u003e for pearl millet.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eImpact of various amendments on pigments\u003c/h2\u003e \u003cp\u003eThe data reveals that treatment with contaminated soil (K), without the addition of BC, substantially (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) diminished chlorophyll a, b, levels and total chlorophyll (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). However, the addition of both tested BC led to a significant increase in chlorophyll content. The most substantial enhancements in chlorophyll content, specifically a 41% and 28% increase for chl. a, a 43% and 51% increase for chl. b, and a 32% and 29% increase in total chlorophyll were observed at a BC application rate of 1.5% for B1 and B2, respectively \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eSeveral studies conducted by \u003cb\u003eBashir et al. (2018), Mehmood et al. (2018)\u003c/b\u003e, and \u003cb\u003eHelaoui et al. (2023)\u003c/b\u003e have reported that the application of BC derived from rice straw can elevate chlorophyll content and mitigate oxidative stress. Conversely, \u003cb\u003eBelhaj et al. (2016)\u003c/b\u003e noted a decrease in chlorophyll a and b due to increased HM contents in soil contaminated by mining activities. Additionally, BC appeared to alleviate the stress caused by HMs on sorghum plants through mechanisms such as toxic metal adsorption onto the BC surface (Arabyarmohammadi et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), enhancement of soil physico-chemical properties, and enhanced nutrient uptake by sorghum (\u003cb\u003eOk et al., 2015; Younis et al., 2016\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eFurthermore, \u003cb\u003eShahbaz et al. (2018)\u003c/b\u003e identified a substantial negative association between nickel concentration in the shoots and sunflower and maize plants as well as their physiological characteristics after incorporating BC. This result indicates that BC may accelerate the adsorption of HMs (through surface interactions), ultimately improving the photosynthetic capacity of the tested plants under HM stress.\u003c/p\u003e \u003cp\u003eConversely, compared to the K treatment, CA application alone elevated chl. a, b, and total chlorophyll by 86%, 82%, and 81%, respectively. In comparison, the application of VW alone increased these chlorophyll levels by 79%, 58%, and 68%, respectively, in plants subjected to HM stress. However, the most significant increases in chl. a, b, and total chlorophyll, at 105%, 80%, and 91%, respectively, were detected in plants grown (under the combined VW and CA application) \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e Afshan et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) illustrated that CA application significantly increased photosynthetic pigment levels in \u003cem\u003eBrassica napus\u003c/em\u003e when grown in soil contaminated by Cr, compared to exposure to Cr alone. This increase in photosynthetic pigments may be attributed to enhanced antioxidant enzyme activity, which reduces the production of malondialdehyde (MDA) and electrolyte leakage. These findings are consistent with those of Mallhi et al. (2019), who found that photosynthetic pigments of castor plants grown in Pb-contaminated soil significantly enhanced after applying CA compared to treatments with Pb alone. The beneficial role of CA in the photosynthetic system of plants exposed to heavy metal stress has been described in numerous studies, including \u003cb\u003eShakoor et al. (2014)\u003c/b\u003e, Afshan et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), and \u003cb\u003eFarid et al. (2017)\u003c/b\u003e. Applying CA enhances essential nutrients\u0026rsquo; uptake and photosynthetic pigments\u0026rsquo; formation (\u003cb\u003eFarid et al., 2017\u003c/b\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eImpact of various amendments on HMs concentration and uptake\u003c/h2\u003e \u003cp\u003eThe addition of BC led to improved plant growth and alleviated the tested HM mobility and availability within the plant tissues. The substantial decline (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) in HM bioavailable concentration in soil induced the decreased translocation in the root and shoots of sorghum (grown in contaminated soil). For instance, when comparing the contents of Zn, Pb, Ni, Mn, Fe, Cu, and Cd in sorghum plants\u0026rsquo; roots, the application of B1 and B2 reduced them from 3.10, 129.29, 170.61, 54.07, 7.96, 47.91, and 115.59 mg/kg to 2.66, 115.08, 149.86, 45.62, 7.19, 39.79, and 103.80 mg/kg, respectively for B1, and 2.66, 116.69, 142.39, 45.70, 7.10, 37.89, and 103.86 mg/kg, respectively for B2 (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cb\u003eConcentration of Cd, Cu, Fe, Mn, Ni, Pb, and Zn (mg/Kg) on roots and shoots of sorghum plants at different treatments.\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"7\" nameend=\"c8\" namest=\"c2\"\u003e \u003cp\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003eRoot\u003c/span\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCd\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePb\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e 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\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e149.86\u0026thinsp;\u0026plusmn;\u0026thinsp;6.42\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e45.62\u0026thinsp;\u0026plusmn;\u0026thinsp;2.67\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e39.79\u0026thinsp;\u0026plusmn;\u0026thinsp;2.60\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e103.80\u0026thinsp;\u0026plusmn;\u0026thinsp;4.14\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e 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\u003cp\u003e45.70\u0026thinsp;\u0026plusmn;\u0026thinsp;3.28\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.10\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e37.89\u0026thinsp;\u0026plusmn;\u0026thinsp;3.90\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e103.86\u0026thinsp;\u0026plusmn;\u0026thinsp;4.27\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB2\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e116.18\u0026thinsp;\u0026plusmn;\u0026thinsp;4.24\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e140.79\u0026thinsp;\u0026plusmn;\u0026thinsp;5.13\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e46.66\u0026thinsp;\u0026plusmn;\u0026thinsp;3.08\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.87\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2.79\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e102.07\u0026thinsp;\u0026plusmn;\u0026thinsp;5.28\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e142.92\u0026thinsp;\u0026plusmn;\u0026thinsp;6.36\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e187.15\u0026thinsp;\u0026plusmn;\u0026thinsp;8.07\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e64.32\u0026thinsp;\u0026plusmn;\u0026thinsp;3.28\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.28\u0026thinsp;\u0026plusmn;\u0026thinsp;1.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e53.77\u0026thinsp;\u0026plusmn;\u0026thinsp;2.77\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e131.69\u0026thinsp;\u0026plusmn;\u0026thinsp;6.36\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCA\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e144.77\u0026thinsp;\u0026plusmn;\u0026thinsp;5.64\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e184.81\u0026thinsp;\u0026plusmn;\u0026thinsp;6.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e65.36\u0026thinsp;\u0026plusmn;\u0026thinsp;3.31\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.64\u0026thinsp;\u0026plusmn;\u0026thinsp;1.24\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e56.91\u0026thinsp;\u0026plusmn;\u0026thinsp;3.87\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e138.38\u0026thinsp;\u0026plusmn;\u0026thinsp;4.76\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cspan type=\"BoldUnderline\" class=\"BoldUnderline\" name=\"Emphasis\"\u003eShoot\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eCd\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eCu\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eFe\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eMn\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eNi\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003ePb\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003eZn\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eK\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.41\u0026thinsp;\u0026plusmn;\u0026thinsp;2.82\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e38.75\u0026thinsp;\u0026plusmn;\u0026thinsp;3.25\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e9.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8.83\u0026thinsp;\u0026plusmn;\u0026thinsp;1.10\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e20.78\u0026thinsp;\u0026plusmn;\u0026thinsp;2.32\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eVW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.54\u0026thinsp;\u0026plusmn;\u0026thinsp;2.07\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e39.87\u0026thinsp;\u0026plusmn;\u0026thinsp;3.45\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.99\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e20.44\u0026thinsp;\u0026plusmn;\u0026thinsp;1.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e22.95\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e34.89\u0026thinsp;\u0026plusmn;\u0026thinsp;2.19\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e17.98\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB1\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.09\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e35.02\u0026thinsp;\u0026plusmn;\u0026thinsp;3.22\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.66\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.19\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e18.22\u0026thinsp;\u0026plusmn;\u0026thinsp;2.08\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.04\u0026thinsp;\u0026plusmn;\u0026thinsp;1.20\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e32.85\u0026thinsp;\u0026plusmn;\u0026thinsp;2.87\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.09\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e18.14\u0026thinsp;\u0026plusmn;\u0026thinsp;2.10\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eB2\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.10\u0026thinsp;\u0026plusmn;\u0026thinsp;2.10\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e33.82\u0026thinsp;\u0026plusmn;\u0026thinsp;3.26\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e18.50\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCA\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.79\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e45.87\u0026thinsp;\u0026plusmn;\u0026thinsp;2.35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11.20\u0026thinsp;\u0026plusmn;\u0026thinsp;2.15 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e10.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.44\u0026thinsp;\u0026plusmn;\u0026thinsp;2.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCA\u0026thinsp;+\u0026thinsp;VW\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e29.37\u0026thinsp;\u0026plusmn;\u0026thinsp;1.65\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.51\u0026thinsp;\u0026plusmn;\u0026thinsp;2.86ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.02\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e11.07\u0026thinsp;\u0026plusmn;\u0026thinsp;1.22\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e26.09\u0026thinsp;\u0026plusmn;\u0026thinsp;1.79\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eAll the data are presented as mean values of three independent experiments (n\u0026thinsp;=\u0026thinsp;3). K represents un-amendment (control), VW (addition of vermiwash), B1 (biochar date palm), B1\u0026thinsp;+\u0026thinsp;VW (biochar date palm\u0026thinsp;+\u0026thinsp;vermiwash), B2 (biochar Prosopis), B2\u0026thinsp;+\u0026thinsp;VW (biochar Prosopis\u0026thinsp;+\u0026thinsp;vermiwash), CA (citric acid), and CA\u0026thinsp;+\u0026thinsp;VW (citric acid\u0026thinsp;+\u0026thinsp;vermiwash). All the means sharing common letter(s) are insignificantly different at P\u0026thinsp;\u0026le;\u0026thinsp;0.05.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFurthermore, it was observed that both B1 and B2 treatments significantly reduced the concentrations of Cd, Cu, Fe, Mn, Ni, Pb, and Zn in the shoots compared to the control soil. These reductions amounted to 30.88%, 13.10%, 9.96%, 12.65%, 20.81%, 17.55%, and 13.47%, respectively for B1, and 29.41%, 8.97%, 15.23%, 10.89%, 22.97%, 19.71%, and 12.70%, respectively for B2. The results demonstrate that applying BC immobilized HMs such as Cd, Cu, Fe, Mn, Ni, Pb, and Zn in the soil reduces their concentrations in the roots and shoots of sorghum plants. Numerous studies have previously reported similar findings, where BC amendments in heavy metal-contaminated soil resulted in the immobilization of these metals, leading to decreased uptake by plants (\u003cb\u003eMoon et al., 2013; Rehman et al., 2018; Yin et al., 2016; Gasc\u0026oacute; et al., 2019; Helaoui et al., 2023\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eIn a study by Abbas et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), varying rice straw BC levels (0, 1.5, 3.0, and 5% w/w) were applied to Cd-contaminated soil. The outcomes indicated that BC treatments lowered Ni and Cd concentrations in wheat grains, roots, and shoots compared to controls. In addition, Cd concentration in wheat grains declined by 26%, 42%, and 57% following the application of 1.5%, 3.0%, and 5.0% BC, respectively, compared with the control. The reduction in HM concentrations in the roots of sorghum plants may be attributed to the selectivity of plants and metals\u0026rsquo; immobilization within the soil, as evidenced by a decrease in DTPA-extracted soil HM levels (\u003cb\u003eLu et al., 2012\u003c/b\u003e). Functional groups in the BC may have immobilized Cu and Pb concentrations (\u003cb\u003eUchimiya et al., 2011\u003c/b\u003e). Immobilization of HMs through BC applications has been previously documented (Ahmad et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; \u003cb\u003eJones et al., 2016; Ok et al., 2015\u003c/b\u003e). Consequently, applying both tested BCs (B1 and B2) proved more effective in promoting plant growth and reducing metal concentrations in the roots and shoots of sorghum plants (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e.\u003c/p\u003e \u003cp\u003eHowever, the application of 1.5 mM CA to the plants increased the Cd, Cu, Fe, Mn, Ni, Pb, and Zn contents in the roots by 22.31%, 9.54%, 8.84%, 15.94%, 14.22%, 10.90%, and 12.23%, respectively, whereas in the shoots, increased by 12.26%, 13.66%, 15.52%, 18.84%, 11.90%, 17.17%, and 18.32%, respectively, compared to the plants grown in K treatment (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). A comprehensive comprehension of the accumulation of HMs and the transportation behavior of plants is crucial in the process of selecting suitable agents for phytoremediation or enhancing plant stress tolerance. Organic acids have been widely recognized for their dual function in facilitating metal accumulation and acting as allies in the development of stress tolerance in plants involving metal tolerance (\u003cb\u003eEhsan et al., 2014; Mahmud et al., 2017\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eExogenous application of CA has been shown to enhance HM uptake in numerous plants (\u003cb\u003eNajeeb et al., 2011;\u003c/b\u003e Almaroai et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; \u003cb\u003eEhsan et al., 2014\u003c/b\u003e). CA in this study significantly increased the HM content in the shoots and roots of sorghum, confirming its potential for phytoremediation. The CA application also considerably elevated HM translocation from roots to shoots.\u003c/p\u003e \u003cp\u003eApplying VW alone and combined with BC 1\u0026amp; 2 and CA has no significant effect on tested HM contents compared to treatments with the tested BC and chelating agents alone. The findings of this study provide evidence in favor of the hypothesis that the effectiveness of phytoextraction can be enhanced by augmenting the dry mass and, to a lesser degree, the metal accumulation in the upper parts of the plants (Table\u0026nbsp;\u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eChelating agents are frequently employed to improve the availability of metals and promote their accumulation in plants. However, this increased metal uptake can lead to decreased plant biomass due to the toxicity associated with excessive metal absorption. Therefore, the simultaneous application of VW and CA represents an optimal approach to address this issue. Moreover, this combination can enhance a plant\u0026rsquo;s inherent antioxidative defense mechanisms, as suggested by \u003cb\u003eGao et al. (2010)\u003c/b\u003e.\u003c/p\u003e \u003cp\u003eConversely, the efficacy of the phytoremediation procedure, encompassing phytoextraction and phytostabilization, is contingent upon the quantity of biomass produced by the roots and shoots, as well as their capacity to absorb HMs. As a result, the quantification of metal absorption by the roots and shoots of sorghum plants was computed and graphically depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The findings of the study indicated a significant increase (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) in the uptake of heavy metals (HMs) by both the shoot and root when exposed to all tested treatments, as compared to the untreated soil potassium (K). The combined application of calcium (CA) and vermicompost (VW) resulted in the highest uptake of heavy metals (HM) in both the shoot and root. This observation can be attributed to the increased dry matter production associated with the application of VW, as compared to K. The concurrent utilization of CA and VW resulted in a significant enhancement in plants\u0026rsquo; absorption of various heavy metals. Specifically, the uptake of Cd increased from 4.37 to 11.28 \u0026micro;g/pot, Cu increased from 163.48 to 408.66 \u0026micro;g/pot, Fe increased from 219.82 to 535.79 \u0026micro;g/pot, Mn increased from 66.32 to 179.42 \u0026micro;g/pot, Ni increased from 11.82 to 32.27 \u0026micro;g/pot, lead (Pb) increased from 59.58 to 159.53 \u0026micro;g/pot, and Zn increased from 143.19 to 385.95 \u0026micro;g/pot, respectively, in comparison to the control treatment using K.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eHowever, applying BC and VW improved the sorghum plant\u0026rsquo;s phytoextraction ability by enhancing plant growth. This study\u0026rsquo;s findings indicate that using BC in conjunction with nutrient supplements may be a viable approach for enhancing the phytoremediation of heavy metals (HMs) when employing sorghum as a bioenergy crop. The results obtained in this study were consistent with the findings reported by \u003cb\u003eHouben et al. (2013), Fellet et al. (2014), and Houben et al. (2013)\u003c/b\u003e also observed the phytostabilization of cadmium (Cd) and zinc (Zn) by Brassica napus plants when BC was applied. A study conducted by Fellet et al. (2014) observed that the application of BC led to increased uptake of lead (Pb) by various plant species. This phenomenon was attributed to plant growth promotion, thereby indicating the potential for phytostabilization. The application of BC frequently leads to enhanced soil water-holding capacity, nutrient retention, microbial community composition, cation exchange capacity (CEC), and plant responses to diseases. The enhanced attributes of the BC ultimately lead to a higher crop output, indicating the BC\u0026rsquo;s potential as a phytoextractant (\u003cb\u003eGraber et al., 2010; Paz Ferreiro et al., 2014\u003c/b\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003ePhytoextraction efficiency of metals\u003c/h2\u003e \u003cp\u003eThe bioconcentration factors (BCF) and translocation factors (TF) for sorghum are presented in Figs.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e\u0026amp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e. Baker (1981) has classified BCF into three distinct categories, which are as follows: Plants exhibiting a BCF below 1 are classified as excluders, while those falling within the range of 1 to 10 are classified as accumulators. Plants with BCF values exceeding 10 are referred to as hyperaccumulators. The HM BCF values for the sorghum plant were observed to follow a decreasing order, with Cd having the highest average value, followed by Cu and Fe, and then Mn, Ni, Pb, and Zn. The mean BCF value across all treatments was found to be below 1 \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe study findings revealed that the transfer factor (TF) and bioconcentration factor (BCF) values for the various amendments examined were found to be less than one. This suggests that the process of phytoextraction is not viable for the sorghum plants in the present investigation despite their elevated biomass yield (\u003cb\u003eSteve and Zhao, 2003\u003c/b\u003e). Metal accumulation by the shoot and root of sorghum plants is depicted in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The study results indicated that a considerable proportion of the metals was accumulated in the roots, as opposed to the shoots. The results presented in \u003cb\u003e(\u003c/b\u003eFigs.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e\u0026amp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e indicate a decline in the translocation of HMs in the roots, as evidenced by the lower values of TF and BCF. This finding suggests that the phytostabilization process was not as effective in preventing the accumulation of HMs in the roots. Phytostabilization refers to the capacity of certain plant species to effectively accumulate a substantial amount of HMs in their root systems, thereby mitigating the risk of contamination in nearby or underground water sources \u003cb\u003e(Vamerali et al., 2009).\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003eFellet et al. (2014) and Karami et al. (2011)\u003c/b\u003e have documented that BC application has demonstrated suitability for the phytostabilization of certain HMs. Therefore, based on our findings, it can be suggested that the utilization of date palm and prosobis BC demonstrates effective potential for the immobilization of heavy metals in soil. However, the implementation of phytoextraction using sorghum plants in soil contaminated by mining activities is not considered a viable option. According to \u003cb\u003eGoswami and Das (2015\u003c/b\u003e), soluble chemical elements have the ability to enter roots through two distinct pathways: the apoplastic pathway, which involves movement through the cell wall-free space, and the symplastic pathway, involving transportation across the plasma membrane of root cells followed by movement through the cytoplasm.\u003c/p\u003e \u003cp\u003eMultiple studies have demonstrated that roots serve as a protective barrier against the translocation of HMs, thereby protecting the stems and other aboveground plant components from contamination and minimizing oxidative stress \u003cb\u003e(Panwar et al., 2002; Liu et al., 2019).\u003c/b\u003e The observed variations in HM concentrations in different plant parts indicate the presence of distinct cellular mechanisms responsible for partitioning, translocating, and bioaccumulating these metals within plant systems \u003cb\u003e(Sinha et al., 2007; Sharma and Dietz, 2009).\u003c/b\u003e Furthermore, HM translocation and uptake to the shoots are inherently connected to the speciation of soil organic matter, soil pH, SOM, and elements (\u003cb\u003eKabata-Pendias, 2010\u003c/b\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eMultivariate Analysis\u003c/h2\u003e \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e \u003ch2\u003eA principal component analysis (PCA)\u003c/h2\u003e \u003cp\u003eIt has been performed to evaluate the effect of different treatments in removing tested HMs (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). Metals are correlated negatively with PCA components. In contrast, other metals correlate positively with PCA components. It was shown that some treatments are suitable for removing HMs; on the other hand, some are not suitable according to PCA analysis. B1, B2, B1\u0026thinsp;+\u0026thinsp;VW, and B2\u0026thinsp;+\u0026thinsp;VW are negatively correlated with PCA components for removing HMs. Meanwhile, VW, CA, and CA\u0026thinsp;+\u0026thinsp;VW are positively correlated with PCA for removing HMs. The most critical treatment used in this study to remove HMs is Vermiwash (VW), which is effectively necessary for removing Zn. The results indicated that Cu and Zn are correlated positively with PCA. In comparison, Cd and Ni correlated negatively with PCA components (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e). PCA 1 (91.82%) and PCA 2 (5.5%) explain the majority of the data, so there is no need for the other PCA components.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003ePhytoextraction is highly dependent on potentially toxic metals\u0026rsquo; bioavailability in soil and plant characteristics. Remediation of soil contaminated with Cd, Cu, Fe, Mn, Ni, Pb, and Zn using both tested BC (data palm and Prosopysis), citric abid, and VW separately and together were investigated by performing a pot experiment for 90 days. The BC application significantly decreased the bioavailable tested HM contents, while the application of CA significantly enhanced the bioavailability of treated HMs in mine-contaminated soil. Compared to unamendment treatment (contaminated soil without any treatment), applying different treatments (BC, CA, and VW) separately and together led to significant enhancement of morphological traits and chlorophyll content of sorghum plants. Moreover, HM contents and uptake to sorghum shoot and root substantially (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) elevated with all tested treatments compared to the untreated soil (K).\u003c/p\u003e \u003cp\u003eFurthermore, all tested amendments\u0026rsquo; TF and BCF values are less than unity. This finding suggests that phytoextraction is not feasible for the sorghum plants in the current study despite higher biomass production. In contrast, VW application has been found to alleviate the detrimental impacts of HMs on both plant growth and photosynthetic efficiency. Additionally, when co-inoculated with contaminated soil, VW has been observed to enhance the capacity for phytoremediation. However, additional research is required to verify the results reproducibility and assess the potential for practical implementation in the field.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of data and materials:\u0026nbsp;\u003c/strong\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e: Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate:\u003c/strong\u003e Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to publish\u003c/strong\u003e: Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests:\u003c/strong\u003e The authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution statement:\u003c/strong\u003e Hanan E. Osman and Ruwaydah Fadhlallah conceived and designed, and performed the experiments; Hanan E. Osman; Ruwaydah S. Fadhlallah and Mohamed H. El-Morsy interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e The authors would like to express their gratitude to the Deanship for Research \u0026amp; Innovation, Ministry of Education in Saudi Arabia, for providing financial support for this research project under project number IFP22UQU4320730DSR030.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003e\u003cstrong\u003eAbbas T, Rizwan M, Ali S, Zia-ur-Rehman M, Qayyum MF, Abbas F, Hannan F, Rinklebe J , Ok YS (2017)\u003c/strong\u003e Effect of biochar on cadmium bioavailability and uptake in wheat (\u003cem\u003eTriticum aestivum L.)\u003c/em\u003e grown in a soil with aged contamination. 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Sci Total Environ 571:206\u0026ndash;217.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"environmental-science-and-pollution-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"espr","sideBox":"Learn more about [Environmental Science and Pollution Research](https://www.springer.com/journal/11356)","snPcode":"11356","submissionUrl":"https://submission.nature.com/new-submission/11356/3","title":"Environmental Science and Pollution Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Mahad AD'Dahab mine contaminated soil, biochar, cirtic acid, phytoremediation, sorghum, vermiwash","lastPublishedDoi":"10.21203/rs.3.rs-3445202/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3445202/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePhytoremediation technology is an eco-friendly technology for the treatment of a polluted environment. In contrast, it has been demonstrated that both natural and synthetic amendments can enhance the process of phytoremediation of heavy metals (HMs) from polluted soils through the utilization of bioenergy crops. This work assessed the synergistic impact of two tested biochar (BC) from data palm (B1) and Prosopis (B2)/ citric acid (CA) and Vermiwash (VW) to enhance phytoremediation of tested HMs (Zn, Pb, Cd, Ni, Cu, Mn, and Fe) from mine-contaminated soil by Sorghum (Sorghum bicolor). The BC and CA amendments alone and combined with VW significantly augmented the proliferation and survival of sorghum grown in mine-contaminated soil.\u003c/p\u003e \u003cp\u003eConsidering the individual and combined applications of VW and BC, the influence on plant growth followed this order: K\u0026thinsp;\u0026gt;\u0026thinsp;VW\u0026thinsp;\u0026gt;\u0026thinsp;B2\u0026thinsp;\u0026gt;\u0026thinsp;B1\u0026thinsp;\u0026gt;\u0026thinsp;B1\u0026thinsp;+\u0026thinsp;VW\u0026thinsp;\u0026gt;\u0026thinsp;B2\u0026thinsp;+\u0026thinsp;VW\u0026thinsp;\u0026gt;\u0026thinsp;CA\u0026thinsp;\u0026gt;\u0026thinsp;CA\u0026thinsp;+\u0026thinsp;VW. Applying tested BC/ CA and VW significantly increased chlorophyll compared to unamended soil. The outcomes revealed a substantial elevation in HM absorption in both shoot and root (p\u0026thinsp;\u0026le;\u0026thinsp;0.05) with all tested treatments compared to the untreated soil (K). The combined application of CA and VW resulted in the highest uptake of HMs in both the root as well as the shoot.\u003c/p\u003e \u003cp\u003eThis study highlights the efficacy of combining CA/BC with VW as a more viable option for remediating mine-contaminated soil compared to individual amendments.\u003c/p\u003e","manuscriptTitle":"Synergistic phytoremediation effect by Sorghum bicolor, citric acid, biochar, and Vermiwash amendment for the remediation of a mine-contaminated soil","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-12-12 15:08:51","doi":"10.21203/rs.3.rs-3445202/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major Revision","date":"2024-03-19T18:12:14+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-01-02T08:53:00+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-12-09T20:26:52+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-11-24T05:28:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"Environmental Science and Pollution Research","date":"2023-11-19T11:04:59+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"environmental-science-and-pollution-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"espr","sideBox":"Learn more about [Environmental Science and Pollution Research](https://www.springer.com/journal/11356)","snPcode":"11356","submissionUrl":"https://submission.nature.com/new-submission/11356/3","title":"Environmental Science and Pollution Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"9f688482-4744-45e4-951b-97666cc64345","owner":[],"postedDate":"December 12th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-06-29T20:56:47+00:00","versionOfRecord":[],"versionCreatedAt":"2023-12-12 15:08:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3445202","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3445202","identity":"rs-3445202","version":["v1"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}