Green synthesis of curcumin nanoparticles, characterization, and their role in alleviating arsenic-induced oxidative stress by enhancing antioxidant defense, photosynthetic pigments, and agronomic traits in wheat.

Green synthesis of curcumin nanoparticles, characterization, and their role in alleviating arsenic-induced oxidative stress by enhancing antioxidant defense, photosynthetic pigments, and agronomic traits in wheat - PMC Skip to main content An official website of the United States government Here's how you know Here's how you know Search Log in Primary site navigation Search Log in Search PMC Full-Text Archive Search in PMC Journal List User Guide As a library, NLM provides access to scientific literature. Inclusion in an NLM database does not imply endorsement of, or agreement with, the contents by NLM or the National Institutes of Health. Learn more: PMC Disclaimer | PMC Copyright Notice Physiol Mol Biol Plants . 2025 Jul 7;31(6):931–958. doi: 10.1007/s12298-025-01615-0 Green synthesis of curcumin nanoparticles, characterization, and their role in alleviating arsenic-induced oxidative stress by enhancing antioxidant defense, photosynthetic pigments, and agronomic traits in wheat Nimra Tahir 1 , Minhas Elahi 1 , Rimsha Aslam 1 , Umar Masood Quraishi 1, ✉ Author information Article notes Copyright and License information 1 Department of Plant Sciences, Faculty of Biological Sciences, Quaid-i-Azam University Islamabad, Islamabad, 45320 Pakistan ✉ Corresponding author. Received 2025 Jan 10; Revised 2025 Jun 2; Accepted 2025 Jun 6; Issue date 2025 Jun. © Prof. H.S. Srivastava Foundation for Science and Society 2025. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. PMC Copyright notice PMCID: PMC12314167  PMID: 40756435 Abstract This study evaluates the potential of green-synthesized curcumin nanoparticles (Cur-NPs) for mitigating arsenic (As) stress in wheat cultivars Barani-70 and NARC-09. Cur-NPs were characterized by UV–visible spectrophotometry, XRD (36 nm), Fourier Transform Infrared (FTIR) spectroscopy, and Scanning Electron Microscopy (SEM), revealing well-dispersed, amorphous structures and functional groups. Both cultivars were subjected to 10 mg/L arsenic stress and treated with Cur-NPs at 50 mg/L and 100 mg/L through soil and foliar applications. Cur-NPs reduced arsenic uptake by up to 65.01% in leaves and 77.32% in roots. Cur-NP treatments lowered MDA by 50% and H 2 O 2 by 14%. Antioxidant enzyme activities improved; superoxide dismutase (SOD) increased by 13%, peroxidase (POD) by 5%, and catalase (CAT) by 0.5%. Proline content rose by 47%, enhancing osmoprotection. Chlorophyll a and b increased by 24% and 67%, respectively, while carotenoid content rose by 82%. Agronomic traits improved significantly, with plant height increasing by 69.6%, grain yield by 141.3%, and biomass yield by 1260.9%. Starch and total sugar content increased by 155% and 218%, respectively, while protein content rose by up to 225%. Phenolic and flavonoid contents increased by 43% and 37%, strengthening antioxidant defences. These findings underscore the efficacy of Cur-NPs as a sustainable approach to mitigate arsenic toxicity, strengthen antioxidant defence mechanisms, and enhance both physiological traits and agronomic performance in wheat, offering a strong foundation for future field-scale validation and environmental application. Supplementary Information The online version contains supplementary material available at 10.1007/s12298-025-01615-0. Keywords: Curcumin nanoparticles (Cur-NPs), Arsenic stress mitigation, Antioxidant enzyme activity, Heavy metal detoxification, Sustainable agriculture Introduction Agriculture serves as a foundational pillar for economic development worldwide, supporting both developed and emerging economies. As the global population increases, so does the pressure on agricultural systems to meet rising demands for food, energy, and water (Yaqoob et al. 2023 ). This expansion is driven by urbanization and industrialization, which intensify the use of natural resources and place mounting stress on ecosystems (Maja and Ayano 2021 ). One consequence of these pressures is the release of harmful substances, including pesticides, fertilizers, and heavy metals, into the environment. Industrial activities, mining, and unsustainable farming practices contribute to the contamination of agricultural soils, compromising plant health and reducing crop yields (Ali et al. 2019 ). Among environmental toxicants, heavy metals are particularly concerning due to their persistence and tendency to accumulate in the food chain, posing ecological and health risks (Bhunia 2017 ; Alengebawy et al. 2021 ). Heavy metal contamination, in particular, is a major environmental issue impacting crop productivity (Li et al. 2024 ). Heavy metals (HMs), such as cadmium, lead, and arsenic, enter agricultural soils through various routes including industrial effluents, fossil fuel combustion, sewage irrigation, and the overuse of chemical inputs like pesticides and fertilizers (Singh et al. 2021 ; Abd Elnabi et al. 2023 ; Hafeez et al. 2023 ). Natural processes such as erosion and volcanic activity also contribute to their distribution(Hafeez et al. 2023 ). Once present in soils, HMs are absorbed by plants, where they interfere with metabolic functions, affect nutrient dynamics, and ultimately threaten human and animal health via bioaccumulation (Chen et al. 2018 ). While several heavy metals contribute to agricultural stress, arsenic (As) is particularly harmful due to its high toxicity and mobility. Arsenic contamination has been linked to reduced crop productivity, compromised seed germination, and disruption of photosynthesis (Ramezani et al. 2021 ; Li et al. 2023 ). Arsenic contamination in agricultural areas has escalated through both human-induced and natural sources, resulting in its accumulation in soils and eventual uptake by crops (Ghorbani et al. 2021 ). In plants, arsenic induces oxidative damage by generating reactive oxygen species (ROS) such as singlet oxygen, hydroperoxyl radicals, and hydrogen peroxide (H 2 O 2 ), which disrupt normal cellular activities and contribute to cellular stress and toxicity (Ghorbani et al. 2024 ). Arsenic stress directly affects key crops like wheat ( Triticum aestivum L. ), which is one of the most widely cultivated and consumed grains worldwide (Saeed et al. 2022 ). Its ability to absorb and accumulate arsenic in grains raises significant concerns about food safety. Research has reported that consumption of arsenic-contaminated wheat poses potential carcinogenic risks to human health, with exposure levels in some regions exceeding international safety standards (Suman et al. 2020 ; Saeed et al. 2021 ). These challenges highlight the urgent need for efficient and sustainable remediation strategies that can be applied at the agricultural scale. Recent developments in nanotechnology suggest that nanoscale materials could play a significant role in alleviating metal-induced stress in plants (Sen et al. 2025 ; Sol-Magdaleno et al. 2025 ). Nano-remediation—using engineered nanomaterials to immobilize or transform contaminants—offers promising advantages such as higher reactivity, targeted delivery, and environmental compatibility (Ekrami et al. 2022 ; Ahmad et al. 2023 ). Green synthesis of nanomaterials, such as carbon nanoparticles, nanofibers, nanocomposites, and metal-based nanoparticles, offers an eco-friendly alternative to conventional chemical and physical remediation methods. Green-synthesized nanoparticles are favoured for their environmental compatibility, reproducibility, and scalability (Rivera-Solís et al. 2024 ). Among these materials, green-synthesized nanoparticles are gaining attention due to their non-toxic and environmentally friendly production processes (Abd El-Hack et al. 2021 ). Curcumin nanoparticles (Cur-NPs), derived from the bioactive compound in turmeric ( Curcuma longa ), exhibit strong antioxidant and chelating properties that may help reduce heavy metal stress in plants (Rahmani et al. 2017 ). Cur has already been reported to modulate metal toxicity and limit bioaccumulation in biological systems (Smirnova et al. 2023 ; Zhang et al. 2024 ). Despite growing awareness of arsenic’s phytotoxicity and its implications for food safety, practical and sustainable strategies for mitigating arsenic stress in crops remain limited. Conventional remediation methods are often costly, environmentally invasive, or inefficient under field conditions. Green nanotechnology, although promising, remains underutilized in this domain. In particular, curcumin nanoparticles (Cur-NPs)—with their metal-chelating ability, antioxidative capacity, and plant-compatible profile—have not been thoroughly investigated for arsenic stress mitigation in cereal crops. This study aims to address this knowledge gap by evaluating the effectiveness of green-synthesized Cur-NPs in improving wheat tolerance to arsenic contamination. This study aims to address this knowledge gap by evaluating the effectiveness of green-synthesized Cur-NPs in improving wheat tolerance to arsenic contamination. The hypothesis is that Cur-NPs will reduce arsenic uptake, mitigate arsenic-induced oxidative stress, and improve overall plant health and yield. The novelty of this work lies in the use of green-synthesized Cur-NPs as a sustainable bioremediation strategy. Curcumin's antioxidant and metal-chelating properties at the nanoscale present an environmentally friendly approach to address heavy metal toxicity in agriculture. The findings could provide valuable insights into enhancing crop robustness and promoting food security in arsenic-affected regions, advancing sustainable agricultural practices. Therefore, this study aims to evaluate the effectiveness of green-synthesized curcumin nanoparticles in alleviating arsenic-induced stress in wheat through physiological, biochemical, and agronomic assessments. Materials and methods Green synthesis and physical characterization of curcumin NPs Formation of Curcuma longa ( C. longa ) extract Curcuma longa rhizomes were obtained from local vendors and thoroughly washed to remove mud particles and impurities (Supplementary Fig. 5 ). They were dried in an oven at 70 °C for 72 h. The dried turmeric rhizomes were ground into a fine powder, which was sieved and used for further studies. To produce the extract, 20 g of C. longa powder was mixed in 400 ml of distilled water in a 500 ml flask and blended for 1 h at 40 °C on a hot plate with a magnetic stirrer. After stirring, the extract was placed in a water bath for incubation at 80 °C for 1 h. The resulting mixture underwent filtration three times using a muslin cloth and then Whatman filter paper. This method was adapted from (Matinise et al. 2018 ). Green synthesis of curcumin NPs For the green synthesis of Cur-NPs, previously utilized protocols of (Iqbal et al. 2019 ) with minor modifications were followed (Supplementary Fig. 5 ). 0.1 M solution of manganese chloride (MnCl 2 ) was prepared and stirred on hot plate at 400 °C for 1 h. To produce Cur-NPs, a purified extract solution of C. longa rhizome was gradually added to the MnCl 2 solution with ratio 1:1. The mixture was heated to 80 °C while being stirred at 500 rpm for 2 h. The settled pellet of Cur-NPs was obtained following centrifugation at 3000 rpm for 25 min and rinsed with methanol (three times). The precipitated Cur-NPs pellet was calcinated and subsequently characterized using various spectroscopic techniques (Jayarambabu et al. 2020 ). Characterization of curcumin nanoparticles UV–visible absorption spectrophotometer of Cur-NPs (Cur-NPs) The formation of Cur-NPs was verified using a UV-400 UV–visible spectrophotometer (Germany) within a wavelength range of 200–800 nm. FT-IR spectroscopy of green Cur-NPs The identification of functional groups associated with plant extract-stabilized Cur-NPs was performed using FTIR spectroscopy (Alpha, Bruker, Germany). For this analysis, the KBr pellet technique was employed. Specifically, 10 mg of nanoparticle powder was thoroughly mixed with 100 mg of potassium bromide (KBr) and compressed to form a pellet. The FTIR spectrum was recorded across a wavenumber range of 400–4000 cm −1 at a resolution of 4 cm −1 , following the method described by Ali et al. ( 2022a , b ). XRD analysis of synthesized CUR-NPs To assess the crystalline structure of the synthesized Cur-NPs, approximately 10 mg of nanoparticle powder was subjected to X-ray diffraction (XRD) analysis (Esther Arland and Kumar 2024 ). The analysis was conducted using a PANalytical X-ray diffractometer (Netherlands) equipped with a Cu Kα radiation source, operating at a current of 30 mA and a voltage of 50 kV. The diffraction patterns were recorded over a 2θ angular range of 20° to 80°, with a scan rate of 13 s per increment, and the measurements were performed at a constant temperature of 25 °C. The corresponding size was determined using the Scherrer equation: where ‘D’ represents the crystalline size, ‘k’ is the shape constant (0.9), and ‘λ’ is the wavelength of the X-rays, which is 1.5 × 10 −10 m. SEM and EDX analysis of CUR-NPs The morphological characteristics, particle size, and elemental composition of the synthesized green nanoparticles (Cur-NPs) were examined using SEM and EDX. To prepare the sample, Cur-NPs were dispersed in distilled water and subjected to ultrasonic treatment for 30 min. A drop of this suspension was placed on a conductive carbon-coated tape and allowed to air dry. The sample was then analyzed using the VEGA3 TESCAN instrument, providing detailed imaging and elemental analysis (Ali et al. 2022a , b ). Invitro biocompatible studies Stimulatory and inhibitory bioassay of Cur-NPs The phytotoxic impact of green-synthesized Cur-NPs was assessed using the radish seed assay method (RSA) (Uddin et al. 2021 ). Different concentrations (0, 30, 60, 120, 250, 500, 1000 µl/mL) of Cur-NPs were added to each Petri dish containing sterilized filter paper (Whatman filter paper) and six radish seeds were placed at equal distances. Following incubation, various indices were calculated, including final germination, percentage inhibition, and the number of days (T50) needed to achieve 50% germination of the total seed count (Pérez-García and González-Benito 2006 ). Ultimately, the length of the seedlings was assessed in cm. Antihaemolytic activity The Bioethical Committee of Quaid-i-Azam University, Islamabad (Pakistan), provided research clearance (#BEC-FBS-QAU2019-143) for the use of human blood, which was collected in EDTA vials containing 3 ml of blood from healthy individuals. Phosphate buffer saline PBS solution was used to dilute the human blood before spinning it for 12 min at 3000 rpm. The pellet was then washed three times using 0.2 M PBS. To measure the absorbance, 1 mL of diluted erythrocyte suspension, 0.5 mL PBS, and 1 mL of each extract (300, 200, and 100 µL/mL) were incubated for 1 h at 36 °C. After that, the mixture was centrifuged for 15 min at 3000 rpm. In this experiment, Triton X-100 served as the positive control. The negative control was Dimethyl Sulfoxide (DMSO). The following formula was used to compute the percentage of inhibition once the absorbance of the supernatant was recorded at 540 nm (Okoli et al. 2008 ). Anti-microbial analysis of Cur-NPs Antifungal activity of Cur-NPs to control Aspergillus flavus on peanut The seeds of Arachis hypogaea (peanut) were procured from a nearby market. The cotyledons underwent disinfection using 2% (v/v) hydrogen peroxide for 10 min, followed by a rinse with tap water. They were pre-treated with Cur-NPs at concentrations of 100, 500, and 1000 µL/mL for 2 h, followed by inoculation with an A. flavus conidia suspension (1 × 10 6 conidia/mL) as described by Yao et al. ( 2017 ). The A. flavus conidia suspension was prepared using SDA media (potato extract, peptone, dextrose). A. flavus was added to the SDA media and incubated at 28 °C for 7 days. After incubation, the A. flavus suspension was prepared by mixing the media. The treated peanuts were subsequently incubated at 28 °C for 7 days, after which the cotyledons infected with A. flavus were collected. As a control, A. flavus was replaced with sterile double-distilled water (d.d.H 2 O). Each treatment was conducted in triplicate, using five cotyledons for each replicate, and the experiment was repeated twice (Zhang et al. 2023 ). Experimental design The experiment was conducted in PVC pipes to compare normal arsenic contamination levels and the role of curcumin nanoparticles in mitigating arsenic stress. For this experiment, two wheat cultivars, Barani-70 and NARC-09, were selected based on their differential sensitivity to arsenic stress. Barani-70 has been reported as arsenic-tolerant, while NARC-09 is considered arsenic-sensitive, according to prior screening and literature (Saeed et al. 2024 ) The PVC pipes were filled with 2 kg of a soil-sand mixture (in a 2:1 ratio). Random soil samples were analyzed according to the protocols established by Safdar et al. ( 2020a , b ). The soil used in the experiment contained sandy-loam texture, with 71.9% sand, 12.7% silt, and 15.4% clay, a pH of 7.8, electrical conductivity (EC) of 2.9 dS/m, TDS value of 310 mg/kg, and 0.68% organic matter. The experiment was performed under controlled conditions in a semi-outdoor environment. Average day/night temperatures ranged between 25 and 28 °C during the experiment, with relative humidity maintained at approximately 60–65%. Plants were exposed to natural sunlight (light intensity ~ 1200 to 1500 µmol m −2 s −1 ) with a natural photoperiod of ~ 12 h light/12 h dark. No rainfall occurred during the experimental period, and water was provided through manual irrigation. Arsenic stress (10 mg/kg using sodium meta-arsenite, Sigma Aldrich, St. Louis, MO, USA) was imposed on the 7th day after germination in the designated stress groups (Anas et al. 2024a , b , c ). The experimental design of the following treatments is shown in Table 1 . Two concentrations of Cur-NPs, 50 mg/L and 100 mg/L, were selected based on previous studies reporting their effectiveness in enhancing stress tolerance in plants under metal-contaminated conditions (Hamed et al. 2025 ; Jamil Abbasi et al. 2025 ). Table 1. Summary of treatment conditions applied to wheat cultivars under different curcumin nanoparticle concentrations and arsenic stress levels Code Cultivar Treatment description Cur-NPs concentration Stress condition Application method C1V1 Barani-70 Control (no Cur-NPs, no arsenic stress) None No arsenic stress – C1V2 NARC-09 Control (no Cur-NPs, no arsenic stress) None No arsenic stress – C2V1 Barani-70 Cur-NPs only (no arsenic) 50 or 100 mg/L No arsenic stress Soil C2V2 NARC-09 Cur-NPs only (no arsenic) 50 or 100 mg/L No arsenic stress Soil C3V1 Barani-70 Arsenic stress only (no Cur-NPs) None Arsenic (10 mg/kg) – C3V2 NARC-09 Arsenic stress only (no Cur-NPs) None Arsenic (10 mg/kg) – T1V1 Barani-70 Cur-NPs treatment (50 mg/L) without arsenic stress 50 mg/L No arsenic stress Foliar spray T1V2 NARC-09 Cur-NPs treatment (50 mg/L) without arsenic stress 50 mg/L No arsenic stress Foliar spray T2V1 Barani-70 Cur-NPs treatment (100 mg/L) without arsenic stress 100 mg/L No arsenic stress Foliar spray T2V2 NARC-09 Cur-NPs treatment (100 mg/L) without arsenic stress 100 mg/L No arsenic stress Foliar spray T3V1 Barani-70 Cur-NPs treatment (50 mg/L) without arsenic stress 50 mg/L No arsenic stress Soil application T3V2 NARC-09 Cur-NPs treatment (50 mg/L) without arsenic stress 50 mg/L No arsenic stress Soil application T4V1 Barani-70 Cur-NPs treatment (100 mg/L) without arsenic stress 100 mg/L No arsenic stress Soil application T4V2 NARC-09 Cur-NPs treatment (100 mg/L) without arsenic stress 100 mg/L No arsenic stress Soil application T5V1 Barani-70 Cur-NPs treatment (50 mg/L) under arsenic stress 50 mg/L Arsenic (10 mg/kg) Soil T5V2 NARC-09 Cur-NPs treatment (50 mg/L) under arsenic stress 50 mg/L Arsenic (10 mg/kg) Soil T6V1 Barani-70 Cur-NPs treatment (100 mg/L) under arsenic stress 100 mg/L Arsenic (10 mg/kg) Soil T6V2 NARC-09 Cur-NPs treatment (100 mg/L) under arsenic stress 100 mg/L Arsenic (10 mg/kg) Soil Open in a new tab Before sowing, the seeds were surface-sterilized with 2% hydrogen peroxide (H 2 O 2 ) for 5–10 min, followed by thorough washing with distilled water. After sterilization, the seeds were planted in PVC pipes. Upon harvesting, several phenotypic and agronomic traits were evaluated using the protocol of Safdar et al. ( 2020a , b ). Various parameters such as fresh weight (FW), shoot length (SL), and leaf area (LA), leaf area (LA), plant height (PH), tiller number (TN), spikelets per spike (SpS), thousand kernel weight (TKW), biological yield (BY), and harvest index (HI) were measured. Grain yield (GY) was determined by harvesting the grains from each experimental unit (PVC pipe) at maturity, and the total grain weight was recorded and expressed as grams per square meter (g m −2 ), following the method of Ain et al. ( 2015 ). Chlorophyll contents were measured after germination and at heading stage. The chlorophyll meter, SPAD-502 Konica Minolta sensing Inc., was used to measure the chlorophyll content. Root samples were collected after harvesting and thoroughly washed to remove any adhering soil particles. Images of the roots were captured from a fixed distance of 60 cm using a digital camera. Root morphological traits, including total root length, number of median roots, number of root tips, average root orientation, average root diameter, and root length-to-diameter ratio, were analyzed using Rhizo–Vision Explorer software (Anas et al. 2023 ). Average root diameter was calculated automatically by the software based on pixel-based measurements from the entire scanned root structure (Supplementary Fig. 6 ). Antioxidant enzyme activity assay Enzymatic antioxidant activity was measured using a microplate-based spectrophotometric method. To assess superoxide dismutase (SOD) activity, a 3 mL reaction solution was assembled with 0.1 mL of 0.2 M methionine, 0.01 mL of 2.25 mM nitro blue tetrazolium (NBT), 0.01 mL of 3 mM EDTA, 1.5 mL of 0.1 M potassium phosphate buffer at pH 7.0, 0.05 mL of crude enzyme extract, and 1 mL of distilled water. The reaction was initiated by adding 0.1 mL of 60 µM riboflavin. Absorbance was recorded at 560 nm, and SOD activity (EC 1.15.11.1) was expressed in units per gram of fresh weight (U g −1 FW). (Bufarwa et al. 2024 ). For the determination of POD (EC 1.11.1.7), a reaction mixture consisting of 2.2 mL of 0.1 M potassium phosphate buffer (pH 6.0), 0.5 mL of 0.018 mM guaiacol, 0.2 mL of 30% hydrogen peroxide (H 2 O 2 ), and 0.1 mL of enzyme extract. After that the absorbance was recorded at 420 nm, POD (EC 1.11.1.7) activity was measured and represented as U/g/FW (Anas et al. 2024a , b , c ). Catalase activity was determined using an assay mixture composed of enzyme extract, 100 mM potassium phosphate buffer (pH 7.0), and 75 mM hydrogen peroxide. The reaction rate was monitored by measuring the decrease in absorbance at 240 nm with a UV–visible spectrophotometer. One unit of catalase activity was defined as the amount of enzyme required to decompose 1 µmol of H 2 O 2 per minute per milligram of protein. After measuring the absorbance. of CAT at 240 nm, the CAT(EC 1.11.1.6) enzyme activity was represented as Unit g −1 FW (Venkidasamy et al. 2019 ). Oxidative stress markers The method of Gharib et al. ( 2024 ) was followed to study the generation and concentrations of H 2 O 2 in the leaves and roots of both cultivars. Hydrogen peroxide content was quantified from fresh leaf samples. A 0.5 g sample of leaf tissue was homogenized in 5 mL of 0.1% (w/v) trichloroacetic acid on ice. The homogenate was centrifuged at 6000 rpm for 30 min at 4 °C. To the collected supernatant, 1.0 mL of 1.0 M potassium iodide (KI) and 0.5 mL of 10 mM potassium phosphate buffer (pH 7.0) were added. Absorbance was measured at 390 nm. In the blank, distilled water was used instead of plant extract. Hydrogen peroxide concentration was calculated from a standard curve and expressed as µmol g −1 fresh weight (FW). Malondialdehyde (MDA) content was determined as an indicator of lipid peroxidation. Fresh leaf tissue (0.1 g) was homogenized in 2 mL of 0.1% trichloroacetic acid (TCA) using a chilled mortar and pestle. The homogenate was centrifuged, and the supernatant was collected. To 1 mL of the supernatant, 4 mL of a reaction mixture containing 0.5% thiobarbituric acid (TBA) in 20% TCA was added. The mixture was then heated in a water bath, cooled, and centrifuged again. Absorbance was recorded at 440 nm, 532 nm, and 600 nm using a UV–visible spectrophotometer. MDA concentration was calculated using standard equations based on absorbance values (Tahir et al. 2024 ). Membrane damage (electrolyte leakage) The permeability of the cell membrane was evaluated using electrolyte leakage, as explained by Bajji et al. ( 2002 ), Omara et al. ( 2022 ). Leaves were collected and washed with deionized water, then placed in test tubes containing 10 ml of deionized water and incubated overnight at 25 °C. Electrical conductivity of the solution (C1) was measured after 24 h. The samples were immersed in a water bath for a duration of 10–15 min and the conductivity reading (C2) was taken after cooling at 25 °C. The electrolyte leakage percentage was measured by following the given formula EL% = C1/C2 × 100. Ascorbate peroxidase (APX) and ascorbic acid oxidase (AOX) analysis The method of Nakano and Asada ( 1981 ) was followed to analyze APX enzyme activity (EC 1.11.1.11). For APX activity, 1 g of sample was ground in 5.0 mL of 0.1 M sodium phosphate buffer (pH 7.0) supplemented with 10% (w/v) polyvinylpyrrolidone (PVP). The homogenate was centrifuged at 12,000 g for 20 min, and the supernatant was collected for further analysis. The reaction mixture contained 0.1 M sodium phosphate buffer (pH 7.0), 0.5 mM ascorbic acid, 0.1 mM hydrogen peroxide (H 2 O 2 ), 0.1 mM EDTA, and the enzyme extract, making a final volume of 700 µL. The decrease in absorbance was recorded at 290 nm for 30 s. APX activity was calculated using an extinction coefficient of 2.8 mM −1 cm −1 and the enzyme's activity was reported as U min −1 g −1 FW. The AOX enzyme analysis at 265 nm wavelength was measured using the method of Ghosh and Biswas ( 2017a , b ) 0.1 g of fresh sample in 5.0 mL of 0.1 M sodium phosphate buffer (pH 7.0). The homogenate was centrifuged at 12,000 g for 20 min, and the resulting supernatant was used for enzyme analysis. The reaction mixture included 3 mL of 0.1 M sodium phosphate buffer (pH 7.0), 1 mL of 0.025% (w/v) ascorbic acid solution, and 1 mL of enzyme extract. Following incubation for 30 min, the reaction was terminated by adding 5 mL of 10% trichloroacetic acid (TCA). The mixture was then titrated against 0.02 M DCPIP solution. AOX activity was result was presented as mg ascorbic acid decomposed mg −1 protein h −1 . Proline assessment Proline content in fresh leaf samples was determined by following the method of Parveen and Siddiqui ( 2021 ) Fresh leaf samples were homogenized in 3% sulphosalicylic acid and then centrifuged to collect the supernatant. Glacial acetic acid and ninhydrin were added to the test tube containing the supernatant. The suspension was left for one hour and then placed in a water bath at 100 °C. The absorbance was measured at the wavelength of 520 nm. L-Proline was used as a reference. Estimation of total chlorophyll and carotenoid The method of Ali et al. ( 2022a , b ) was used to evaluate the amounts of carotenoid and chlorophyll in fresh leaves. To create a fine pulp, 100 mg of fresh leaf sample was crushed using a mortar and pestle with 80% acetone. The mixture was centrifuged at 2500 rpm for ten minutes and supernatant was collected. The absorbance was recorded at 645 and 663 nm for chlorophyll b and chlorophyll a respectively and 480 and 510 nm for carotenoids in comparison to the blank (80% acetone). Estimation of protein content Lowry’s method was used for protein estimation with slight modifications (Morsy 2014 ). A 25 mg sample of dried material was mixed with PBS buffer and centrifuged at 12,000 rpm for 6 min. The supernatant was then used as the test sample, to which Lowry's reagent was added. For each tube, mixed reagent C is prepared by combining 2% Na 2 CO 3 in 0.1N NaOH (Reagent A) and 0.5% CuSO 4 ·5H 2 O in 1% Sodium Potassium Tartrate (Reagent B) in a 50:1 ratio, just before use. This mixture is thoroughly mixed and incubated for 10 min at room temperature. Folin–Ciocalteau reagent is then added, and the solution is allowed to stand for 30 min at room temperature after mixing. The absorbance was measured at 660 nm, and the protein content was reported in mg/g of dry weight (Ramesh et al. 2018 ). Determination of secondary metabolites The total phenolic and flavonoid content of the ethanol extract from the leaves was assessed using the methods described by Mykhailenko et al. ( 2021 ). Extract solutions were combined with an 80/20 (v/v) ethanol/water mixture and incubated for 5 min at room temperature. The total phenolic content was quantified as milligrams of gallic acid equivalent per gram of sample (mg GAE/g). The absorbance was measured at 650 nm. The total flavonoid content was assessed based on the aluminium trichloride method (Kaur et al. 2024 ). The extract solutions were combined with 1 mL of 3% ethanolic AlCl 3 . After a 30-min incubation at room temperature in the dark, the absorbance of the extracts was measured at 510 nm. The total flavonoid content was expressed as milligrams of rutin equivalents per gram of sample (mg RE/g), based on a rutin calibration curve. Estimation of starch The starch content was estimated by the anthrone method (Behera et al. 2024 ). A 0.05 g sample was homogenized in hot 80% ethanol and centrifuged to collect the residue. The residue was repeatedly washed with ethanol to eliminate free sugars. After drying, 3.5 mL of 52% perchloric acid and 2.5 mL of distilled water were added to the residue, and the mixture was left to extract starch for 20 min at 0 °C. Following centrifugation, 0.2 mL of the supernatant was mixed with 0.8 mL of distilled water and 3 mL of freshly prepared anthrone reagent. The reaction mixture was incubated in a water bath at 95 °C for 8 min. Absorbance was measured at 630 nm using a spectrophotometer. The glucose concentration was determined from a standard curve and multiplied by 0.9 to calculate the starch content. Total sugar content estimation Anthrone method was used to determine the total sugar content of the sample (Sarmah et al. 2018 ). A 50 mg portion of dried leaf samples was hydrolyzed with 2.5 N HCl for approximately 3 h in a boiling water bath. Sodium carbonate was then added to neutralize the extract. The extract was centrifuged, and the supernatant was collected. After mixing 1ml of the extract with 2 ml of the anthrone reagent, the mixture was heated in a boiling water bath for 8 min. Absorbance was measured at 630 nm, and the total sugar content was determined by plotting the value against a standard curve of glucose solution. Elemental analysis The As uptake was measured by following the protocol of Sastre et al. ( 2002 ) with slight modification. In 250 ml Pyrex digestion tubes, one gram of material was added, and a pre-digestion step using 8 ml of 70% HNO 3 was run for 16 h at room temperature. After that, the suspension was heated to 250 °C on a hot plate until the brownish vapours became white. After passing through an ashless Whatman filter paper No. 41, the resulting suspension was diluted up to 25 ml of double de-ionized water and placed in plastic bottles for analysis. Statistical analysis The ANOVA results of phenotypic and agronomic traits were performed by using XLSTAT 2024. Scatter plots were generated by using the GGally package, which is an extension to ggplot2, in R version 4.1.4. The paired comparison graphs were made by using Origin pro 2024b software. Results Characterization of curcumin nanoparticles (Cur-NPs) UV–Vis absorption of Cur-NPs The peak values at 199.51 and 262.35 nm indicated the absorption of metal ions while the sharpness of peaks represented stability and well dispersity of the nanoparticles (Fig. 1 B). Fig. 1. Open in a new tab Characterization and anti-haemolytic evaluation of curcumin nanoparticles (Cur-NPs). A FTIR spectrum confirming the presence of functional groups associated with Cur-NPs stabilization. B UV–visible absorption spectrum indicating characteristic peaks related to Cur-NPs stability. C X-ray diffraction (XRD) pattern showing distinct crystalline peaks confirming the crystalline nature of the synthesized nanoparticles. D Anti-haemolytic activity of Cur-NPs at different concentrations (100, 200, and 300 µL/mL) against Triton X-100 induced haemolysis. Error bars represent standard deviation (n = 3) FT-IR spectroscopy of green CUR-NPs The FT-IR spectrum can be seen in (Fig. 1 A). A peak at 3256.697 cm −1 was shown by FTIR analysis of Cur-NPs, indicating the presence of an alcohol group (O–H stretching) on the NPs. The spectra of this sample also showed several distinct bands at 1511.931 cm −1 (N–O stretching) nitro compound and 1736.439 cm −1 (C=O stretching) ester, which indicate the organic components of extracts that are responsible for the effective synthesis of Cur-NPs. XRD analysis of synthesized CUR-NPs The XRD pattern of Cur-NPs showed four peaks (Fig. 1 C). These peaks corresponded to (2θ values) 28.8°, 31.1°, 40.9°, and 50.6° respectively. The Scherrer Equation estimated Cur-NPs average size as 36 nm. This size is within the valid crystalline range. However, SEM analysis revealed an amorphous shape for Cur-NPs. SEM and EDX analysis of Cur-NPs The EDX spectra of NPs showed that silicon (3.54), oxygen (31.01%), and carbon and potassium (47.86%) were the most prevalent elements (Fig. 2 ). SEM imaging revealed that the Cur-NPs exhibited an amorphous morphology. Fig. 2. Open in a new tab A EDS spectrum showing elemental composition (C, O, K, Br, Si) of curcumin nanoparticles. B SEM image depicting surface morphology at 5.00 kx magnification. C Elemental composition table confirming synthesis and stability Phyto stimulatory activity of Cur-NPs Figure 3 shows the percentage increase in seed germination at various concentrations of Cur-NPs (30–1000 µl/mL). The observed percentage stimulation was found to be dose dependent. The growth stimulation of (92%, 150%, and 121%) was observed at concentrations of 30 µl/mL, 120 and 500 µl/mL respectively. Moreover, even at higher concentrations (1000 µl/mL), the seedling’s growth was stimulated up to 435%. Additionally, the relative germination rate (RGR) of seeds was particularly remarkable during the first two days, with germination at 500 µl/mL occurring earlier than in the untreated control. After Day 2, the seeds treated with 500 and 1000 µl/mL exhibited a 3–6% increase in germination compared to the control. In summary, Cur-NPs demonstrated stimulatory effects at certain doses and accelerated the germination process compared to the non-treated control. Moreover, Cur-NPs positively influenced the seedling growth of the radish plant. Fig. 3. Open in a new tab Effect of curcumin nanoparticles (Cur-NPs) on radish seedling growth and germination. A Seedling length (cm) after 2 days under different Cur-NP concentrations. B Germination percentage at 24 h. C Germination percentage at 48 h. D Representative root growth at each concentration with root lengths shown below. Data represent mean ± standard error (n = 3). Different lowercase letters indicate statistically significant differences according to Tukey’s HSD test ( p ≤ 0.05) Antihaemolytic assay The antihaemolytic activity of Cur-NPs was assessed at three different concentrations: 100, 200, and 300 µL/mL. The results indicated that Cur-NPs caused minimal damage to blood cells, with the highest inhibition of 18% recorded at a concentration of 300 µL/mL. Very little inhibition was observed at the lower concentrations of 100 and 200 µL/mL, with values of 16% and 14% respectively, compared to the control (Fig. 1 D). The slight decrease in antihaemolytic activity observed near the DMSO control is attributed to the solvent background rather than any toxic effect of Cur-NPs. Cur-NPs application did not negatively influence plant growth or physiological parameters in wheat, confirming their biocompatibility under the experimental conditions. Antifungal activity of Cur-NPs to control Aspergillus flavus on peanut Cur-NPs demonstrated significant antifungal activity as a disease control agent in peanuts. The highest inhibition rates were observed at concentrations of 500 and 1000 µL/mL, with reductions of 31.8% and 23.9%, respectively. Cur-NPs also showed notable inhibition at 100 µL/mL, with a reduction of 23.3% compared to the control. Additionally, when conidia were collected from the SDA plate and allowed to germinate on curcumin plates, fungal mycelial growth was inhibited on the curcumin plates compared to the control plates (germinated on SDA plates; see Fig. 4 ). Fig. 4. Open in a new tab Antifungal activity of curcumin nanoparticles (Cur-NPs) against fungal growth and spore infection on peanut seeds. A – D Representative images showing fungal growth on Sabouraud Dextrose Agar (SDA) plates treated with control ( A ), 100 µL/mL Cur-NPs ( B ), 500 µL/mL Cur-NPs ( C ), and 1000 µL/mL Cur-NPs ( D ). a – d Visual appearance of peanut seeds exposed to fungal spores under control ( a ), 100 µL/mL Cur-NPs ( b ), 500 µL/mL Cur-NPs ( c ), and 1000 µL/mL Cur-NPs ( d ) treatments Evaluation of phenotypic and agronomic data The analysis of variance showed significant results (< 0.001) among variance phenotypic and agronomic traits as shown in Table 2 and (Fig. 5 ). The application of various treatments demonstrated significant differences in phenotypic traits compared to the arsenic control (C3). Plants treated with 50 mg/L Cur-NPs via foliar spray without arsenic stress (T1V1) exhibited a 69.6% increase in plant height (PH) compared to the arsenic control (C3, arsenic stress only). Similar trends were observed in plants treated with 50 mg/L Cur-NPs via soil application (T3V1) and 100 mg/L Cur-NPs via soil application (T4V1), which showed 63.5% and 67.6% increases, respectively. Grain yield (GY) displayed a positive response to Cur-NP treatments. The highest increase was recorded in plants receiving 100 mg/L Cur-NPs via soil application (T4V1), showing a 141.3% improvement in grain yield. In comparison, 50 mg/L Cur-NPs via foliar spray (T1V1) led to a 107.9% increase in GY, while the lowest increase was observed in T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application), with a 102.8% increase. Biomass yield (BY) followed a similar pattern, with T1V1 showing the highest increase (1260.9%), followed by T3V1 (505.2%). Table 2. Physiological and root morphological parameters of wheat cultivars Barani-70 and NARC-09 under control, arsenic stress, and curcumin nanoparticle (Cur-NP) treatments Variable C1V1 C1V2 C2V1 C2V2 C3V1 C3V2 T1V1 T1V2 T2V1 T2V2 PH (cm) 47 ± 3.53 i 45.03 ± 3.63 i 56.13 ± 2.16 gh 59.7 ± 2.74 efgh 43.3 ± 1.36 i 46.43 ± 1.89 i 73.43 ± 3.65 a 66.76 ± 1.91 abcd 64.96 ± 9.25 cdef 66.63 ± 1.96 abcd LA cm 2 11.63 ± 0.12 gh 10.67 ± 1.04 h 14.32 ± 0.39 cd 12.57 ± 0.34 fg 8.79 ± 0.44i 8.49 ± 0.85 i 15.48 ± 1.28 bc 15.73 ± 0.75 b 13.83 ± 0.59 de 12.90 ± 0.58 ef No. of grain 52 ± 1.41 i 47 ± 3.74 h 53 ± 3.74 i 50 ± 3.741 i 37 ± 3.74 gh 34 ± 3.74 gh 89 ± 1.41 def 55 ± 3.74 i 70 ± 10.19 abdc 58 ± 3.74 fg TKW (g) 33.5 ± 1.47 defg 28.5 ± 1.87 fgh 25.66 ± 1.43 gh 41.33 ± 4.90 cde 31.5 ± 4.89 efg 18.16 ± 2.24 h 65.33 ± 9.13 b 43.66 ± 4.10 cd 48 ± 2.44 c 29.16 ± 3.56 fg GY (g/m 2 ) 0.67 ± 0.02 defg 0.57 ± 0.03 fgh 0.51 ± 0.02 gh 0.82 ± 0.09 cde 0.63 ± 0.09 efg 0.36 ± 0.04 h 1.30 ± 0.18 b 0.87 ± 0.08 cd 0.96 ± 0.04 c 0.58 ± 0.07 fg BY (g/plant) 6.96 ± 2.45 def 3.99 ± 0.13 aef 4.08 ± 0.44 ef 6.16 ± 0.52 def 3.47 ± 0.52 ef 1.86 ± 0.32 f 47.35 ± 13.46 a 13.64 ± 4.15 cde 20.95 ± 10.37 c 8.70 ± 3.39 def CHL. SPAD (spad units) 35.26 ± 2.29 bcde 28.13 ± 3.39 f 35.8 ± 1.91 bcde 40.53 ± 1.81 abc 39.53 ± 6.46 abcd 30.86 ± 5.34 ef 44.16 ± 2.63 a 30.5 ± 1.57 ef 41.06 ± 2.03 ab 34.63 ± 4.06 bcdef Root morphology Variables C1V1 Root C1V2 Root C2V1 Root C2V2 Root C3V1 Root C3V2 Root T1V1 Root T1V2 Root T2V1 Root T2V2 Root MR (Median roots) 6 ± 1 fg 5 ± 1 h 8 ± 1 de 7 ± 1 c 4 ± 1 fg 4 ± 1 e 9 ± 1 d 8 ± 1 c 10 ± 1 fg 9 ± 1 g RT (Number of root tips) 135 ± 7 j 128 ± 6 q 180 ± 9 n 160 ± 8 a 105 ± 6 k 100 ± 5 g 220 ± 10 n 210 ± 9 c 250 ± 12 l 245 ± 11 b ARO (°) 45.2 ± 2.1 a 44.5 ± 1.9 cd 43.9 ± 2.0 de 44.0 ± 1.8 e 41.6 ± 1.7 a 42.1 ± 1.5 b 43.2 ± 1.6 c 43.5 ± 1.7 de 42.8 ± 1.8 c 42.9 ± 1.6 h RLD (cm/cm 3 ) 1.75 ± 0.12 r 1.68 ± 0.11 p 2.25 ± 0.15 n 2.05 ± 0.14 h 1.30 ± 0.09 o 1.20 ± 0.08 f 2.75 ± 0.18 c 2.60 ± 0.17 m 3.10 ± 0.20 e 3.00 ± 0.19 q AD (mm) 0.42 ± 0.02 j 0.40 ± 0.01 a 0.47 ± 0.03 i 0.45 ± 0.02 q 0.38 ± 0.01 e 0.37 ± 0.01 h 0.50 ± 0.02 o 0.48 ± 0.02 n 0.52 ± 0.03 c 0.51 ± 0.02 k RL (cm) 65.8 ± 4.1 c 62.4 ± 3.7 m 78.2 ± 5.6 a 72.7 ± 5.2 g 50.4 ± 3.9 d 48.5 ± 3.5 k 85.9 ± 6.2 j 82.3 ± 5.9 q 90.4 ± 7.1 n 88.7 ± 6.7 b Variable T3V1 T3V2 T4V1 T4V2 T5V1 T5V2 T6V1 T6V2 P*T PH (cm) 70.8 ± 3.96 abc 69.43 ± 1.73 abc 72.56 ± 0.82 ab 66.16 ± 1.43 bcde 58.8 ± 2.45 fgh 60.5 ± 3.341 defgh 55.1 ± 3.06 h 62.4 ± 2.56 defg *** LA cm 2 18.57 ± 0.35 a 19.15 ± 0.43 a 19.11 ± 0.52 a 16.48 ± 0.22 b 13.38 ± 0.08 def 13.36 ± 0.05 def 13.93 ± 0.46 de 15.44 ± 0.30 bc *** No. of grain 67 ± 3.74 abc 86 ± 2.82 ab 83 ± 3.74 ab 53 ± 3.74 ghi 52 ± 3.74 ghi 50 ± 3.74 59 ± 3.74 61 ± 3.74 *** TKW (g) 62.5 ± 6.53 b 65 ± 10.00 b 76 ± 3.89 a 66.83 ± 10.81 ab 43.83 ± 1.69 cd 34.5 ± 3.08 defg 50 ± 2.94 c 36.66 ± 2.77 def *** GY (g/m 2 ) 1.25 ± 0.13 b 1.3 ± 0.20 b 1.52 ± 0.07 a 1.33 ± 0.21 ab 0.87 ± 0.033d efg 0.69 ± 0.06 c 1 ± 0.02 c 0.73 ± 0.05 def *** BY (g/plant) 21.057 ± 6.61 c 21.18 ± 3.7 c 32.37 ± 11.14 b 17.1 ± 3.85 cd 8.84 ± 2.23 def 5.16 ± 0.47 ef 8.88 ± 1.05 def 6.74 ± 0.93 def *** CHL. SPAD (spad units) 33.5 ± 4.42 def 35.8 ± 3.82 bcde 40 ± 2.00 abcd 34.33 ± 3.10 cdef 39.76 ± 1.73 abcd 40.36 ± 0.60 abc 36.63 ± 3.55 bcde 35.46 ± 3.16 bcde *** Root morphology Variables T3V1 Root T3V2 Root T4V1 Root T4V2 Root T5V1 Root T5V2 Root T6V1 Root T6V2 Root P*T MR (Median roots) 7 ± 1 fg 7 ± 1 f 8 ± 1fg 7 ± 1 fg 7 ± 1 e 6 ± 1 b 9 ± 1 fg 8 ± 1 a ** RT (Number of root tips) 170 ± 8 h 165 ± 8 f 185 ± 9 o 180 ± 9 p 160 ± 8 i 155 ± 7 d 215 ± 10 m 210 ± 10 e ns ARO (°) 44.1 ± 2.0 b 44.2 ± 1.8 de 43.7 ± 1.9 fg 43.8 ± 1.7 gh 44.5 ± 2.0 ef 44.8 ± 1.9 cd 43.0 ± 1.8 a 43.1 ± 1.6 b ** RLD (cm/cm 3 ) 2.20 ± 0.14 k 2.05 ± 0.13 j 2.35 ± 0.15 g 2.25 ± 0.14 l 2.00 ± 0.13 i 1.95 ± 0.13 d 2.70 ± 0.18 b 2.60 ± 0.17 a ns AD (mm) 0.46 ± 0.02 f 0.45 ± 0.02 d 0.47 ± 0.02 b 0.46 ± 0.02 g 0.44 ± 0.02 m 0.43 ± 0.02 p 0.49 ± 0.02 l 0.48 ± 0.02 r * RL (cm) 75.2 ± 5.5 l 70.9 ± 5.1 p 78.5 ± 5.7 e 76.2 ± 5.4 h 70.5 ± 5.1 i 68.7 ± 4.9 f 82.5 ± 6.1 o 80.8 ± 5.9 l *** Open in a new tab Values represent mean ± standard deviation (n = 3). Statistical significance among treatments was determined by one-way ANOVA followed by Tukey’s HSD test (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, non-significant results are denoted as “ns”. P*T = Probability between treatments). The measured physiological parameters include PH (plant height, cm), LA (leaf area, cm 2 ), GY (grain yield, g/m 2 ), BY (biomass yield, g/plant), TKW (thousand kernel weight, g), SpS (spikelets per spike), and CHL SPAD (chlorophyll content measured in SPAD units). The assessed root morphological parameters include RL (root length, cm), RLD (root length density, cm/cm3), AD (average diameter, mm), RT (number of root tips), ARO (average root orientation, degrees), and MR (median roots, number). Treatment abbreviations (e.g., C1V1, T1V1, etc.) are described in Table 1 . Physiological traits were recorded at the heading stage, while root traits were evaluated after the final harvest. Arsenic stress (10 mg/kg) was imposed 7 days after germination Fig. 5. Open in a new tab Correlation matrix illustrating Pearson correlation coefficients between parameters. Positive correlations are shown in red and negative correlations in blue, with color intensity reflecting correlation strength Leaf area (LA) was also positively affected by Cur-NP treatments. Plants in T3V1 (50 mg/L Cur-NPs, soil application) showed a 98.4% increase in LA, while those in T4V1 (100 mg/L Cur-NPs, soil application) demonstrated a 141.3% increase. Thousand kernel weight (TKW) significantly improved under Cur-NP treatments, with T3V2 (50 mg/L Cur-NPs via soil) achieving a 125.3% increase, and T4V1 (100 mg/L Cur-NPs via soil) showing a 117.5% increase, reflecting enhanced grain filling and productivity. Interestingly, in treatments without arsenic stress but with Cur-NPs (C2V1 and C2V2), a slight reduction in agronomic traits was observed. This decrease may be attributed to mild physiological stress induced by nanoparticles in the absence of arsenic. Root morphology also exhibited substantial improvements. Plants in T1V1 (50 mg/L Cur-NPs via foliar spray) displayed a 40% increase in median root length and a 76.2% increase in root diameter, suggesting enhanced root development. Antioxidant enzyme activity The enzymatic antioxidants SOD, POD, and CAT in the two cultivars against various treatments responded differently to arsenic (As) stress as shown in Table 3 (Supplementary Fig. 1 ). In the leaf tissues, in leaf tissues, CAT activity generally decreased across most treatments. The most pronounced reductions were recorded in plants exposed to 100 mg/L Cur-NPs via soil without arsenic stress (T4V1, − 2.00%) and 100 mg/L Cur-NPs under arsenic stress via soil application (T6V2, − 2.03%). Conversely, POD activity exhibited significant increases in several treatments. Plants treated with 50 mg/L Cur-NPs via foliar spray without arsenic stress (T1V2) showed the highest increase (86.69%), while those receiving 100 mg/L Cur-NPs via foliar spray without arsenic stress (T2V2) demonstrated a 49.04% increase. SOD activity followed a similar trend, with the highest improvements recorded in T4V2 (100 mg/L Cur-NPs via soil, + 9.37%) and T1V2 (50 mg/L Cur-NPs via foliar spray, + 9.37%). Soil-applied treatments, such as T4V1 (100 mg/L Cur-NPs) and T3V2 (50 mg/L Cur-NPs), also showed positive SOD responses, with 9.14% and 8.12% increases, respectively. Table 3. Antioxidant enzyme activities in shoot and root tissues of wheat cultivars Barani-70 and NARC-09 under control, arsenic stress, and curcumin nanoparticle (Cur-NP) treatments Variable C1V1 C1V2 C2V1 C2V2 C3V1 C3V2 T1V1 T1V2 T2V1 T2V2 CAT(U/mg protein) 0.36 ± 0.0017 ab 0.36 ± 0.0001 ab 0.36 ± 0.0001 ab 0.35 ± 0.0008 efg 0.35 ± 0.001 cde 0.36 ± 0.0003 bcd 0.35 ± 0.0012 ef 0.35 ± 0.0007 de 0.36 ± 0.0017 abc 0.36 ± 0.0012 a POD (U/mg protein) 4.52 ± 0.72 abcd 2.63 ± 0 f 2.88 ± 0.03 ef 4.17 ± 0.43 abcde 5.4 ± 0.08 a 5.49 ± 0.01 a 5.02 ± 0.26 ab 4.91 ± 2.49 abc 3.81 ± 0.44 bcdef 3.92 ± 0.58 bcdef SOD (U/mg protein) 1.64 ± 0.0077 def 1.6 ± 0.0045 f 1.63 ± 0.0058 ef 1.64 ± 0.0105 def 1.69 ± 0.0321 cd 1.67 ± 0.0138 de 1.74 ± 0.0089 abc 1.75 ± 0.0105 ab 1.77 ± 0.0045 ab 1.67 ± 0.0674 de Antioxidants root Variable C1V1 C1V2 C2V1 C2V2 C3V1 C3V2 T1V1 T1V2 T2V1 T2V2 CAT (U/mg protein) 0.35 ± 0.0006 ab 0.35 ± 0.0002 cde 0.35 ± 0.0016 bc 0.35 ± 0.0002 abc 0.35 ± 0.0001 abc 0.35 ± 0.0001 ab 0.35 ± 0.0006 ef 0.35 ± 0.0004 def 0.35 ± 0.0026 def 0.35 ± 0.0001 def POD (U/mg protein) 4.13 ± 0.82 bcde 3.04 ± 0 g 3.04 ± 0 g 3.33 ± 0.4 fg 4.3 ± 0 abc 4.29 ± 0 abcd 4.52 ± 0.42 bcdef 3.6 ± 0.48 defg 3.42 ± 0.12 fg 3.97 ± 0.4 bcdef SOD (U/mg protein) 1.73 ± 0.0061 cd 1.69 ± 0.0045 d 1.73 ± 0.0045 c 1.72 ± 0.015 cd 1.73 ± 0.0029 cd 1.71 ± 0.0058 cd 1.78 ± 0.0058 ab 1.79 ± 0.0118 ab 1.8 ± 0.0228 ab 1.78 ± 0.0094 ab Variable T3V1 T3V2 T4V1 T4V2 T5V1 T5V2 T6V1 T6V2 P*T CAT(U/mg protein) 0.35 ± 0.0003 cde 0.35 ± 0.0002 efg 0.35 ± 0.0008 fgh 0.35 ± 0.0037 efg 0.35 ± 0.0015 h 0.35 ± 0.0037 ef 0.35 ± 0.0004 gh 0.35 ± 0.0004 h *** POD (U/mg protein) 3.43 ± 0.2 def 3.49 ± 0.1 cdef 3.73 ± 0.01 bcdef 3.73 ± 0.63 bcdef 4.07 ± 0.58 abcdef 3.84 ± 0.57 bcdef 4.52 ± 0.53 cdef 5.03 ± 0.45 ab *** SOD (U/mg protein) 1.66 ± 0.0607 de 1.73 ± 0.0121 bc 1.79 ± 0.0045 a 1.75 ± 0.0369 ab 1.76 ± 0.0061 ab 1.73 ± 0.0058 bc 1.74 ± 0.0077 bc 1.73 ± 0.0045 bc *** Antioxidants root Variable T3V1 T3V2 T4V1 T4V2 T5V1 T5V2 T6V1 T6V2 P*T CAT (U/mg protein) 0.35 ± 0.0006 def 0.35 ± 0.0009 f 0.35 ± 0.0006 ef 0.35 ± 0.0006 bcd 0.35 ± 0.0026 bc 0.36 ± 0.0018 a 0.35 ± 0.0008 ab 0.35 ± 0.0036 f *** POD (U/mg protein) 3.47 ± 0.03 efg 3.66 ± 0.04 cdefg 4.6 ± 0.61 ab 3.76 ± 0.47 cdef 4.89 ± 0.04 a 4.22 ± 0.14 abcd 4.48 ± 0.35 ab 4.26 ± 0.08 abcd *** SOD (U/mg protein) 1.77 ± 0.0442 b 1.8 ± 0.0058 ab 1.78 ± 0.0029 ab 1.73 ± 0.0448 c 1.77 ± 0.0034 b 1.8 ± 0.0045 ab 1.77 ± 0.0134 ab 1.81 ± 0.0089 a *** Open in a new tab Values represent mean ± standard deviation (n = 3). Statistical significance among treatments was determined by one-way ANOVA followed by Tukey’s HSD test (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, Non-significant results are denoted as “ns”. P*T = Probability between treatments). The measured biochemical parameters include CAT (catalase activity, U/mg protein), POD (peroxidase activity, U/mg protein), and SOD (superoxide dismutase activity, U/mg protein Despite these improvements, some treatments resulted in reduced POD activity. Plants treated with 50 mg/L Cur-NPs via soil (T3V1) and those exposed to 50 mg/L Cur-NPs under arsenic stress via soil (T5V1 and T5V2) experienced declines, with T5V2 showing the most significant reduction (− 30.05%). This suggests that the concentration and application method of Cur-NPs can directly influence enzyme responses. In root tissues, CAT activity remained relatively stable across treatments, with minor fluctuations. Slight increases were observed in plants treated with 100 mg/L Cur-NPs under arsenic stress via soil (T6V1, + 0.19%) and 50 mg/L Cur-NPs under arsenic stress via soil (T5V2, + 0.51%). POD activity predominantly increased in roots, particularly in plants treated with 100 mg/L Cur-NPs via soil (T2V2, + 30.59%) and 50 mg/L Cur-NPs via foliar spray (T4V2, + 23.68%). SOD activity in root tissues also showed consistent positive trends, with the highest increases recorded in T3V2 (50 mg/L Cur-NPs via soil, + 6.50%) and T6V2 (100 mg/L Cur-NPs under arsenic stress, + 5.85%) (Table 4 ). Table 4. Photosynthetic pigments and secondary metabolite contents in wheat cultivars Barani-70 and NARC-09 under control, arsenic stress, and curcumin nanoparticle (Cur-NP) treatments Variable C1V1 C1V2 C2V1 C2V2 C3V1 C3V2 T1V1 T1V2 T2V1 T2V2 CHL. A (mg/g fresh weight) 5.42 ± 0.27 bcde 6.86 ± 0.81 abcd 3.75 ± 1.65 ef 2.12 ± 0.42 f 3.84 ± 0.47 def 3.75 ± 1.49 ef 5.18 ± 1.48 bcde 3.77 ± 1.56 ef 4.56 ± 0.9 bcdef 4.85 ± 0.32 bcdef CHL.B (mg/g fresh weight) 4.77 ± 0.26 bcdef 6.06 ± 0.43 abc 4.41 ± 0.63 bcdef 4.89 ± 0.73 bcde 3.74 ± 0.42 ef 2.95 ± 0.8 f 5.83 ± 0.24 abcd 3.61 ± 1 ef 3.98 ± 0.53 def 3.71 ± 0.62 ef T.CHL (mg/g fresh weight) 4.17 ± 0.23 bcde 4.17 ± 0.37 ab 4.98 ± 0.54 abc 5.53 ± 0.64 ab 4.01 ± 0.37 bcde 3.04 ± 0.69 e 5.1 ± 0.21 ab 3.16 ± 0.87 de 4.7 ± 0.46 abdce 3.26 ± 0.55 de CARO (mg/g fresh weight) 27.48 ± 4.15 cdef 42.88 ± 6.7 a 27.42 ± 5.3 cdef 30.67 ± 8.26 bcdef 20.78 ± 2.5 f 25.8 ± 3.72 def 43.3 ± 4.28 a 30.83 ± 7.43 bcdef 28.87 ± 0.97 bcdef 31.87 ± 0.8 bcde FLAVENOID (mg/g fresh weight) 0.58 ± 0.02 b 1.1 ± 0.01 ef 1.06 ± 0.05 fg 1.14 ± 0.01 def 1.06 ± 0.03 fg 0.88 ± 0.09 g 1.22 ± 0.12 def 1.21 ± 0.08 def 1.28 ± 0.04 cde 1.18 ± 0.04 def PHENOL (mg/g fresh weight) 0.55 ± 0.05 hi 0.6 ± 0.03 fgh 0.76 ± 0.04 bc 0.59 ± 0.01 fgh 0.44 ± 0.03 j 0.5 ± 0.03 ij 0.78 ± 0.08 b 0.65 ± 0.03 defg 0.71 ± 0.06 bcd 0.86 ± 0.04 a Variable T3V1 T3V2 T4V1 T4V2 T5V1 T5V2 T6V1 T6V2 P*T CHL. A (mg/g fresh weight) 3.88 ± 1.24 def 4.55 ± 2.07 cdef 4.77 ± 2.53 abc 4.77 ± 0.27 a 4.77 ± 0.61 ab 4.45 ± 1.08 def 4.77 ± 3.92 abcde 4.38 ± 1.23 def *** CHL.B (mg/g fresh weight) 4.3 ± 0.6 cdef 4.43 ± 1.2 bcdef 6.11 ± 1.49 abc 6.91 ± 1.01 a 6.27 ± 0.32 ab 3.58 ± 0.85 ef 5.34 ± 2.71 abcde 3.9 ± 1 def *** T.CHL (mg/g fresh weight) 4.77 ± 0.52 abcd 3.88 ± 1.04 bcde 5.34 ± 1.29 ab 6.03 ± 0.88 a 5.47 ± 0.27 ab 3.14 ± 0.74 de 4.67 ± 0.23 abcde 3.41 ± 0.87 cde *** CARO (mg/g fresh weight) 34.45 ± 5.25 abcd 27.53 ± 11.82 cdef 39 ± 3.41 ab 42.9 ± 3.23 a 37.98 ± 2.85 abc 26.85 ± 4.56 def 23.18 ± 17.68 ef 28.3 ± 3.36 bcdef *** FLAVENOID (mg/g fresh weight) 1.65 ± 0.21 a 1.47 ± 0.1 abc 1.3 ± 0.14 cd 1.19 ± 0.06 def 1.46 ± 0.08 bc 1.21 ± 0.03 def 1.49 ± 0.14 ab 1.18 ± 0.05 def *** PHENOL (mg/g fresh weight) 0.72 ± 0.02 bcd 0.67 ± 0.05def 0.69 ± 0.01 cde 0.68 ± 0.02 de 0.63 ± 0.03 efg 0.66 ± 0.03 defg 0.59 ± 0.01 gh 0.89 ± 0.01 a *** Open in a new tab Values represent mean ± standard deviation (n = 3). Statistical significance among treatments was determined by one-way ANOVA followed by Tukey’s HSD test (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, Non-significant results are denoted as “ns”. P*T = Probability between treatments). The measured parameters include CHL. A (chlorophyll a content, mg/g fresh weight), CHL. B (chlorophyll b content, mg/g fresh weight), T.CHL (total chlorophyll content, mg/g fresh weight), CARO (carotenoids content, mg/g fresh weight), flavonoid content (mg/g fresh weight), and phenolic content (mg/g fresh weight). Treatment codes (e.g., C1V1, T1V1, etc.) are described in Table 1 Oxidative stress markers In the leaf tissues of the Barani-70 cultivar, exposure to arsenic stress (C3) led to a marked increase in MDA levels as shown in Table 5 (Supplementary Fig. 2 ). However, treatments with curcumin nanoparticles (Cur-NPs) showed a protective effect by reducing MDA concentrations. Specifically, MDA levels in T5V1 (50 mg/L Cur-NPs under arsenic stress, soil application) and T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application) decreased by 45% and 25%, respectively, compared to the arsenic control (C3). Further reductions were observed in T5V2 (50 mg/L Cur-NPs under arsenic stress, soil application) and T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application), where MDA concentrations dropped by 50% and 31.25%, respectively. Treatments without arsenic stress also demonstrated reductions in MDA levels. In T1V1 (50 mg/L Cur-NPs via foliar spray), MDA content decreased by 30%, while T1V2 (50 mg/L Cur-NPs via foliar spray) showed a 50% reduction. Similarly, T2V1 (100 mg/L Cur-NPs via foliar spray) and T2V2 (100 mg/L Cur-NPs via foliar spray) showed MDA reductions of 25% and 56.25%, respectively. Table 5. Leaf antioxidant and stress-related parameters in wheat cultivars subjected to different treatments, including malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ), electrolyte leakage (EL), ascorbate peroxidase (APX) activity, proline content, and ascorbate oxidase (AOX) activity) Variable C1V1 Leaf C1V2 Leaf C2V1 Leaf C2V2 Leaf C3V1 Leaf C3V2 Leaf T1V1 Leaf T1V2 Leaf T2V1 Leaf T2V2 Leaf MDA (µmol/gFW) 0.015 ± 0.003 ab 0.009 ± 0.001 ab 0.011 ± 0.004 ab 0.012 ± 0.004 ab 0.02 ± 0.009 a 0.016 ± 0.002 ab 0.014 ± 0.005 ab 0.008 ± 0.002 b 0.015 ± 0.003 ab 0.007 ± 0.009 b H 2 O 2 (µmol/gFW)) 0.35 ± 0.15 abcd 0.31 ± 0.01 bcd 0.27 ± 0.13 cd 0.25 ± 0.06 d 0.45 ± 0.1 a 0.48 ± 0.02 a 0.41 ± 0.03 ab 0.43 ± 0.06 ab 0.44 ± 0.08 a 0.41 ± 0.18 ab EL (%) 74.34 ± 9.71 bcd 73.29 ± 2.95 bcd 30.95 ± 2.82 fg 36.59 ± 4.9 efg 82.18 ± 2.42 ab 85.77 ± 3.6 a 39.85 ± 11.94 efg 29.8 ± 5.21 g 47.2 ± 9.35 e 31.59 ± 6.61fg APX (U/mg protein) 0.236 ± 0.002 cde 0.236 ± 0.001 cde 0.235 ± 0 e 0.235 ± 0.001 de 0.236 ± 0.002 de 0.236 ± 0.001 cde 0.239 ± 0.001 bcde 0.24 ± 0.001 abcde 0.237 ± 0.001 cde 0.238 ± 0.001 cde PROLINE (µmol/g FW) 0.18 ± 0.14 e 0.33 ± 0.08 de 0.36 ± 0.12 cde 0.41 ± 0.11 bcd 0.43 ± 0.08 bcd 0.44 ± 0.08 bcd 0.44 ± 0.14 bcd 0.47 ± 0.12 bcd 0.48 ± 0.1 abcd 0.46 ± 0.07 abcd AOX (U/mg protein) 3.25 ± 0.05 g 3.29 ± 0.03 g 3.58 ± 0.01 bcd 3.61 ± 0.07 abc 3.5 ± 0.02 ef 3.51 ± 0.03 def 3.55 ± 0.02 cde 3.53 ± 0.03 de 3.48 ± 0.03 ef 3.45 ± 0.01 f Variable T3V1 Leaf T3V2 Leaf T4V1 Leaf T4V2 Leaf T5V1 Leaf T5V2 Leaf T6V1 Leaf T6V2 Leaf P*T MDA (µmol/gFW) 0.009 ± 0.002 b 0.01 ± 0.001 ab 0.01 ± 0.003 ab 0.01 ± 0.001 ab 0.011 ± 0.002 ab 0.008 ± 0.003 b 0.015 ± 0.003 ab 0.011 ± 0.001 ab ns H 2 O 2 (µmol/gFW)) 0.39 ± 0.09 ab 0.43 ± 0.02 a 0.43 ± 0.03 a 0.37 ± 0.06 abc 0.44 ± 0 ab 0.41 ± 0.04 ab 0.41 ± 0.06 ab 0.41 ± 0.1 ab * EL (%) 36.93 ± 4.62 efg 34.01 ± 4.65fg 37.7 ± 1.67 efg 41.4 ± 3.95 ef 69.84 ± 4.99 cd 81.45 ± 2.89 abc 68.25 ± 3.8 cd 64.71 ± 1.16 d *** APX (U/mg protein) 0.237 ± 0.001 cde 0.239 ± 0 bcde 0.237 ± 0.003 cde 0.24 ± 0.001 abcde 0.242 ± 0.002 abc 0.245 ± 0.001 a 0.241 ± 0.003 abcd 0.244 ± 0.002 ab * PROLINE (µmol/g FW) 0.48 ± 0.03 abcd 0.47 ± 0.04 abcd 0.45 ± 0.04 abcd 0.46 ± 0.13 abcd 0.61 ± 0.15 ab 0.65 ± 0.1 a 0.49 ± 0.04 abcd 0.55 ± 0.35 abc * AOX (U/mg protein) 3.53 ± 0.03 de 3.5 ± 0.01 ef 3.52 ± 0 def 3.49 ± 0.05 ef 3.66 ± 0.05 a 3.6249667 ± 0.008807 ab 3.65 ± 0.01 a 3.63 ± 0.01 ab *** Open in a new tab *Significant at p ≤ 0.05, **Significant at p ≤ 0.01, ***Significant at p ≤ 0.001. Non-significant results are denoted as "ns." P*T = Probability between treatments Hydrogen peroxide (H 2 O 2 ) levels also showed a declining trend under Cur-NP treatments. In plants exposed to 50 mg/L Cur-NPs under arsenic stress (T5V1 and T5V2), H 2 O 2 levels decreased by 2% and 14%, respectively, compared to the arsenic control (C3). Similar reductions were recorded in T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application) and T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application), where H 2 O 2 levels decreased by 8% and 14%, respectively. Membrane damage (electrolyte leakage) Arsenic-stressed plants (C3V1 and C3V2) showed the highest electrolyte leakage, with values increasing by 82% and 85%, respectively, compared to the untreated control. This rise reflects severe membrane damage due to oxidative stress induced by arsenic. In contrast, treatments with Cur-NPs reduced electrolyte leakage significantly. In T5V1 (50 mg/L Cur-NPs under arsenic stress, soil application) and T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application), electrolyte leakage decreased by 69% and 68%, respectively, relative to C3. T5V2 (50 mg/L Cur-NPs under arsenic stress, soil application) also demonstrated a 64% reduction in electrolyte leakage. Notably, T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application) showed a slight but consistent reduction compared to C3. Cur-NP treatments without arsenic stress (T1, T2, T3, and T4) also exhibited protective effects, with a significant reduction in electrolyte leakage compared to the control as shown in Table 5 (Supplementary Fig. 2 C). Ascorbate peroxidase (APX) and ascorbic acid oxidase (AOX) analysis The activity of APX was significantly increased in arsenic treated samples as shown in Table 5 (Supplementary Fig. 2 D and E). In leaf tissues of plants exposed to arsenic stress (C3), APX activity increased, but treatments with curcumin nanoparticles (Cur-NPs) further enhanced this response. Specifically, APX activity in T5V1 (50 mg/L Cur-NPs under arsenic stress, soil application) and T5V2 (50 mg/L Cur-NPs under arsenic stress, soil application) showed 2.5% and 3.8% increases, respectively, compared to C3. Similarly, T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application) and T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application) exhibited 2.1% and 3.3% increases, respectively. Ascorbic acid oxidase (AOX) activity showed a similar trend. In leaves of plants treated with Cur-NPs under arsenic stress, AOX activity increased by 4.5% in T5V1 and 3.2% in T5V2. The highest AOX activity was recorded in T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application, + 4.2%) and T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application, + 3.4%). In treatments without arsenic stress, Cur-NP application through foliar spray or soil also resulted in increased AOX activity compared to the control. Proline assessment The level of proline in the leaves of the Barani-70 cultivar increased under nanoparticles treatments as shown in Table 5 (Supplementary Fig. 2 F). In the leaves of the Barani-70 cultivar, proline content increased under arsenic stress but was further enhanced by the application of Cur-NPs. Specifically, T5V1 (50 mg/L Cur-NPs under arsenic stress, soil application) and T5V2 (50 mg/L Cur-NPs under arsenic stress, soil application) exhibited 41% and 47% increases in proline levels, respectively, compared to the arsenic control (C3). In T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application), proline content increased by 25%, while T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application) showed a slight increase. Estimation of total chlorophyll and carotenoid The photosynthetic pigments, including chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll, and carotenoids, displayed significant changes under different treatments (Table 4 , Supplementary Fig. 3 ). Under arsenic stress alone (C3), chlorophyll and carotenoid levels decreased, indicating oxidative damage. However, Cur-NP treatments counteracted these negative effects. In T5V1 (50 mg/L Cur-NPs under arsenic stress, soil application), chlorophyll a content increased by 24%, while carotenoid content surged by 82%. Chlorophyll b and total chlorophyll also showed significant improvements, with increases of 67% and 36%, respectively, compared to C3. In T5V2 (50 mg/L Cur-NPs under arsenic stress, soil application), a similar trend was observed, with Chl a increasing by 18%, Chl b by 21%, total chlorophyll by 3%, and carotenoids by 4%. In T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application), Chl a and Chl b increased by 24% and 42%, respectively, while total chlorophyll and carotenoid levels rose by 16% and 11%. Similarly, T6V2 (100 mg/L Cur-NPs under arsenic stress, soil application) demonstrated a 32% increase in Chl b, along with moderate improvements in Chl a (16%), total chlorophyll (12%), and carotenoids (9%). Estimation of protein content The seed protein content in the Barani-70 and NARC-09 cultivars showed significant variation under different treatments involving curcumin nanoparticles (Cur-NPs) and arsenic stress, as presented in Table 6 (Supplementary Fig. 4 ). In plants treated with 50 mg/L Cur-NPs via foliar spray without arsenic stress, protein content increased by 36% in T1V1 and 50% in T1V2 compared to the arsenic control (C3). Notably, the highest protein content was observed in T2V2 (100 mg/L Cur-NPs via foliar spray), showing a 225% increase, while T2V1 (100 mg/L Cur-NPs via foliar spray) showed a 136% increase. Soil-applied Cur-NPs also enhanced protein content, with T3V1 (50 mg/L Cur-NPs via soil application) and T3V2 (50 mg/L Cur-NPs via soil application) showing 154% and 108% increases, respectively. In contrast, plants exposed to 100 mg/L Cur-NPs via soil application (T4V1 and T4V2) demonstrated 72% and 75% increases in protein content. Under arsenic stress, protein content was lower than in non-stressed treatments. T5V1 (50 mg/L Cur-NPs under arsenic stress, soil application) and T5V2 showed 27% and 8% increases, respectively, while T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application) and T6V2 displayed 54% and 25% increases compared to the arsenic control (C3). Table 6. Biochemical composition of wheat seeds under varying treatments Variable C1V1 C1V2 C2V1 C2V2 C3V1 C3V2 T1V1 T1V2 T2V1 T2V2 PROTEIN (percentage of total weight) 0.13 ± 0.05 c 0.13 ± 0.05 c 0.37 ± 0.05 ab 0.14 ± 0.02 c 0.11 ± 0.03 c 0.12 ± 0.03 c 0.15 ± 0.01 c 0.18 ± 0.02 bc 0.26 ± 0.01 abc 0.39 ± 0.28 a TSC (mg/g FW) 0.98 ± 0.07 efg 1.13 ± 0.06 defg 0.82 ± 0.25 fg 1.1 ± 0.15 defg 0.53 ± 0.12 g 0.53 ± 0.18 fg 3.28 ± 0.31 c 4.48 ± 0.23 b 6.06 ± 0.26 a 5.58 ± 0.71 a STARCH (mg/g FW) 0.58 ± 0.03 fgh 0.53 ± 0.11 gh 0.57 ± 0.08 fgh 0.58 ± 0.05 fgh 0.54 ± 0.05 gh 0.43 ± 0.03 h 1.11 ± 0.13 ab 1.01 ± 0.09 bc 0.88 ± 0.05 cde 0.96 ± 0.05 bcd Variable T3V1 T3V2 T4V1 T4V2 T5V1 T5V2 T6V1 T6V2 P*T PROTEIN (percentage of total weight) 0.28 ± 0.18 abc 0.25 ± 0.07 abc 0.19 ± 0.01 bc 0.21 ± 0.11 abc 0.14 ± 0.04 c 0.13 ± 0.01 c 0.17 ± 0.04 bc 0.15 ± 0 c ns TSC (mg/g FW) 1.91 ± 0.18 d 1.88 ± 0.03 d 4.68 ± 0.33 b 1.67 ± 0.39 de 1.69 ± 0.03 de 1.36 ± 0.14 def 1.204 ± 0.09 defg 1.271 ± 0.22 defg *** STARCH (mg/g FW) 0.95 ± 0.1 bcd 0.77 ± 0.14 def 1.01 ± 0.09 bc 1.13 ± 0.15 ab 0.94 ± 0.09 bcd 0.72 ± 0.04 efg 1.24 ± 0.24 a 1.1 ± 0.11 ab *** Open in a new tab Parameters include protein content, total soluble carbohydrates (TSC), and starch content, reflecting metabolic alterations in response to the treatments. *Significant at p ≤ 0.05, **Significant at p ≤ 0.01, ***Significant at p ≤ 0.001. Non-significant results are denoted as “ns”. P*T = Probability between treatments Determination of secondary metabolites The concentration of secondary metabolites, including flavonoids and phenols, showed notable differences among treatments (Table 4 , Supplementary Fig. 3 E). Flavonoid content significantly increased in T5V1 and T5V2 (50 mg/L Cur-NPs under arsenic stress, soil application), reaching 37% in both treatments. A similar trend was observed in T6V1 (100 mg/L Cur-NPs under arsenic stress, soil application) and T6V2, with increases of 40% and 34%, respectively. Phenolic content also displayed a positive response to Cur-NP treatments. In leaves of plants treated with 50 mg/L Cur-NPs under arsenic stress, phenol levels rose by 43% in T5V1 and 32% in T5V2. The highest phenol content was recorded in T6V2 (100 mg/L Cur-NPs under arsenic stress), which showed a 78% increase, while T6V1 (100 mg/L Cur-NPs under arsenic stress) exhibited a 34% increase. Estimation of starch Starch content in wheat seeds varied significantly under different treatments (Table 6 , Supplementary Fig. 4 B). Plants exposed to 100 mg/L Cur-NPs under arsenic stress (T6V1 and T6V2) showed the highest starch accumulation, with increases of 129% and 155%, respectively, compared to the arsenic control (C3). Treatments with 50 mg/L Cur-NPs under arsenic stress (T5V1 and T5V2) also displayed notable starch content increases, reaching 74% and 67%, respectively. In treatments without arsenic stress, T1V1 (50 mg/L Cur-NPs via foliar spray) and T1V2 exhibited 105% and 134% starch content, while T2V1 (100 mg/L Cur-NPs via foliar spray) and T2V2 showed 62% and 123% starch content, respectively. Soil-applied Cur-NPs also enhanced starch content. T3V1 (50 mg/L Cur-NPs via soil) and T3V2 demonstrated 75% and 79% increases, while T4V1 (100 mg/L Cur-NPs via soil) and T4V2 reached 87% and 162%, respectively. Total sugar content estimation Total sugar content in wheat seeds exhibited significant increases under Cur-NP treatments (Table 6 , Supplementary Fig. 4 C). In arsenic-stressed plants treated with 50 mg/L Cur-NPs (T5V1 and T5V2), sugar content increased by 218% and 156%, respectively. Plants treated with 100 mg/L Cur-NPs under arsenic stress (T6V1 and T6V2) also showed significant increases, with sugar content rising by 127% and 139%, respectively. Interestingly, the arsenic control group (C3) exhibited a marked decrease in total sugar content, with reductions of 45% in C3V1 and 53% in C3V2, reflecting the adverse effects of arsenic stress on sugar accumulation. Elemental analysis In the leaves, a significant reduction in arsenic uptake was observed across all treatments compared to the control (Fig. 6 ). The most significant decrease occurred in T5 for cultivar V2 (65.01%), where 100 mg/L of Cur-NPs were applied with 10 mg/L arsenic stress. The least reduction can be seen in T5 for cultivar V1 (0.43%). In T6V1 about (22.48%) decreased was occur whereas in T6V2 showed (29.76%) decrease in arsenic uptake. Similarly, in the roots, arsenic mitigation displayed greater variability across treatments. The highest reduction was observed in T6 for cultivar V2 (50.74%), which involved soil application of 100 mg/L Cur-NPs under arsenic stress. However, slight decreases in arsenic uptake were noted in certain cases, such as in T6 for cultivar V1 (10.62%) and T5 for cultivar V1 (4.30%). In T5V2 (27.78%) decreased was observed. Fig. 6. Open in a new tab Arsenic (As) accumulation in wheat cultivars Barani-70 and NARC-09 under arsenic stress and curcumin nanoparticle (Cur-NP) treatments. Atomic absorption spectroscopy (AAS) was used to quantify arsenic content in plant tissues. Treatments included arsenic control (C3V1: Barani-70 under arsenic stress, C3V2: NARC-09 under arsenic stress) and arsenic plus Cur-NPs treatments (T5V1: Barani-70 + 50 mg/L Cur-NPs, T5V2: NARC-09 + 50 mg/L Cur-NPs, T6V1: Barani-70 + 100 mg/L Cur-NPs, T6V2: NARC-09 + 100 mg/L Cur-NPs). Error bars represent the standard error of the mean. Significant fluctuations in AA values are observed, with distinct trends between leaf and root samples, highlighting differential elemental absorption and accumulation under varying treatments Discussion The current study demonstrated that curcumin nanoparticles (Cur-NPs) alleviate arsenic (As) stress in bread wheat cultivars (Baraani-70 and NARC-09) by reducing as uptake, restoring essential minerals, alleviating oxidative stress, and improving germination potential. The UV–Vis absorption spectrum of Cur-NPs nanoparticles showed peaks at 199.51 and 262.35 nm, indicating the presence of metal ions, which confirms the formation of Cur-NPs consistent with Pryshchepa et al. ( 2020 ). FTIR of Cur-NPs revealed the presence of several functional groups (Fig. 1 A), such as alcohols, alkanes, alkenes, nitro compounds, carboxylic acids, and esters. These functional groups are indicative of secondary metabolites that act as a natural defence mechanism and provide medicinal benefits to plants that match the previous studies (Uddin et al. 2021 ; Ali et al. 2022a , b ). The XRD peaks observed corresponded to MnO 2 , as Mn served as a carrier agent during the Cur-NPs synthesis (Fig. 1 C). These findings align with Shah et al. ( 2019 ). The use of green-synthesized nanoparticles has demonstrated their potential to enhance plant seedling growth. The findings from this study confirmed the non-toxic nature of Cur-NPs on seedlings, making it a valuable resource for promoting seedling germination. This supports the earlier work of Rasmiya Begum and Jeyawardana ( 2023 ), Vijayaram et al. ( 2024 ), which showed that green-synthesized nanoparticles, derived from various plant parts, are eco-friendly, non-toxic, and promote plant growth, thus serving as a gold standard in this field. The antihaemolytic properties of Cur-NPs displayed a maximum inhibition of 18% at a concentration of 300 µL/mL (Fig. 1 D). These results indicate that, although Cur-NPs do exhibit some ability to prevent haemolysis, their overall efficacy remains modest, even at the highest tested concentration. This moderate activity could be due to the limited bioavailability or interaction of Cur-NPs with blood cells. The current study demonstrated that Cur-NPs effectively restrict the mycelial growth of Aspergillus niger across all tested concentrations (Fig. 4 ). These findings are consistent with the work of Zhang et al. ( 2023 ), which reported that curcumin suppressed both the conidial germination rate and germ tube elongation of Botrytis cinerea. This showed that Cur-NPs exhibits broad-spectrum antifungal activity, potentially acting through similar mechanisms to suppress fungal development in diverse species (Hua et al. 2019 ). The radish seed bioassay and anti-hemolytic analysis collectively affirm the biocompatibility of Cur-NPs, supporting their environmental safety for broader agricultural use (Fig. 3 ). The absence of phytotoxic effects during seed germination and the limited hemolytic activity—observed even at the highest applied concentrations—indicate that Cur-NPs exhibit minimal biological disruption. These findings align with prior studies on bio-based nanomaterials and reinforce the suitability of Cur-NPs for field deployment. The confirmation of low toxicity not only strengthens the ecological relevance of this approach but also addresses key regulatory criteria concerning nanoparticle safety and environmental acceptability, which are critical for advancing nano-enabled technologies in agriculture. The phenotypic and agronomic traits were enhanced by the application of Cur-NPs-compared to the arsenic stress (Table 2 ). Similar results were reported by Farhan et al. ( 2024 ) that the nanoparticles can improve metal stress tolerance and biomass yield of plants. In the current study plant height (PH) was increased significantly as compared to arsenic stress This supports the earlier work of Hammam and Shoala ( 2020 ), where French basil plants treated with Cur-NPs reached heights of 59.00 cm, while the control plants showed corresponding heights of 52.00 cm. Cur-NPs significantly increased the grain yield and biomass yield in arsenic stressed plants. These findings are consistent with the study of Ahmad et al. ( 2020 ), in which ZnO-NPs improved grain and biomass yield of soybean crops against arsenic stress, previous studies have shown that nanoparticles can enhance the leaf area of plants under heavy metal stress. The current research demonstrated that Cur-NPs promoted leaf expansion in plants stressed by arsenic, which is consistent with the findings of Salama et al. ( 2023 ). The significant increase in biomass and grain yield under Cur-NPs treatments highlights their potential for practical applications in arsenic-contaminated soils, addressing food security challenges. The root morphology was improved by using Cur-NPs in As stressed plants, median root length and diameter improved with several treatments. T1V1 led to better root development, showing an increase in both root length and diameter, indicating improved root structure. These findings are related to the previous studies of Upadhyaya et al. ( 2014a , b ). The Thousand Kernel Weight (TKW) significantly increased with T3V2 and T4V1, suggesting better grain filling and higher potential productivity. This indicates that these treatments of Cur-NPs not only improved growth but also enhanced grain weight, a key factor for yield and mitigating the heavy metal stress, aligning with previous studies (Zhou et al. 2021 ) which proved that HMs in the soil can be taken up and altered by the nanoparticles (NPs), thereby decreasing the mobility and bioavailability of heavy metals (HM). Although Cur-NPs generally improved growth parameters under arsenic stress, treatments without arsenic (C2V1 and C2V2) exhibited a slight decline in certain agronomic traits. This is consistent with previous findings where nanoparticle exposure in the absence of metal stress sometimes resulted in mild phytotoxic effects due to oxidative imbalance or nutrient competition (Dimkpa et al. 2012 ; Djanaguiraman et al. 2024 ). Plants exposed to heavy metals produce excessive amounts of reactive oxygen species (ROS), which can result in oxidative stress (Fryzova et al. 2018 ). The resistance to oxidative stress is linked to the activities of ROS-scavenging enzymes such as POD, SOD, and CAT (Hasanuzzaman et al. 2020 ). In this study, a significant change was observed in the SOD levels in wheat cultivars against various treatments (Table 3 ). However, all treatments resulted in an increase in POD activity in both the roots and leaves. An increase in POD activity in wheat varieties under arsenic stress has also been reported by Saeed et al. ( 2023 ). However, a slight increase in CAT levels was observed in the roots of T5V2 and T6V1, whereas a decrease was recorded in the leaves for both treatments. In all other treatments, CAT levels decreased in both the roots and leaves. These findings are similar to Zhang et al. ( 2014 ) where TiO 2 nanoparticles exhibited dual effects on CAT activity: they functioned as an activator at low molar concentrations and as an inhibitor at higher molar concentrations. Cur-NPs not only modulate enzymatic antioxidants like SOD, POD, and CAT but also influence non-enzymatic pathways. The increased activities of key antioxidant enzymes—superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)—in Cur-NPs-treated wheat suggest improved capacity to detoxify reactive oxygen species (ROS) generated under arsenic stress. This biochemical defense likely stabilizes cellular structures and protects key metabolic pathways involved in growth regulation. Such protection is reflected in the observed improvements in morphological traits, particularly root length, shoot height, and biomass accumulation. Efficient ROS scavenging may help maintain cell division and elongation in root meristems and support sustained photosynthetic function in leaves. These findings indicate that enzymatic antioxidant responses play a central role in supporting stress-resilient growth and agronomic performance under arsenic toxicity consistent with the findings of Vasilachi et al. ( 2023 ), Iqbal et al. ( 2024 ). The integration of Cur-NPs potentially stabilizes ROS-scavenging mechanisms, enabling plants to maintain cellular integrity under arsenic stress. The hazardous MDA is one of the end products, and it also acts as an indicator of oxidative stress (Banaee et al. 2024 ). Both cultivars showed reduced MDA levels in their leaves when exposed to As and Cur-NPs, with T5V1 and T6V1, demonstrating a significant decrease compared to T5V2 and T6V2. These results are consistent with earlier research by Faizan et al. ( 2021 ) where exogenous application of ZnO-NPs has decreased the contents of H 2 O 2 and MDA significantly in metal-treated plants that in turn protect the integrity of the cellular membranes. H 2 O 2 induced lipid peroxidation can eventually cause cell death by altering metabolic characteristics Gaschler and Stockwell ( 2017 ). The treatments T5V2 and T6V2 showed decreased levels of H 2 O 2 in the leaves as compared to the control. These findings are consistent with earlier studies, where (Banerjee et al. 2023a , b ) observed a significant increase in endogenous hydrogen peroxide levels in seedlings and plants treated with As. In contrast, the application of varying doses of zinc oxide nanoparticles (ZnO-NPs) resulted in a reduction of hydrogen peroxide (H 2 O 2 ) content in both seedlings and plants. Electrolyte leakage in wheat leaves significantly increased due to arsenic stress. However, the application of Cur-NPs improved the electrolyte leakage in plants affected by arsenic. Our results align with the findings of Banerjee et al. ( 2024 ) who recorded a significant increase in electrolyte leakage (EL) in pea plants exposed to As stress. The application of ZnO nanoparticles notably reduced EL to the lowest levels observed among the various nanoparticle treatments. APX amount increased in the leaf tissues of T5 and T6 treatments under the arsenic stress which is correlated with the studies of Ahmad et al. ( 2020 ) where the supplementation of arsenic-stressed soybean plants with ZnO–NP further enhanced the activity of APX in these plants, showing that ZnO–NP plays a protective role under such challenging conditions. One significant antioxidant enzyme that aids in scavenging ROS like H 2 O 2 is APX. It uses ascorbate as an electron donor to accelerate the transformation of H 2 O 2 into water (H 2 O) (Pandey et al. 2017 ). Our findings similar to previous studies of Farooq et al. ( 2022 ) in which application of MT-Se nanoparticles further enhanced xenobiotic metabolism in Brassica napus by increasing the activity of APX, This suggests that the antioxidant properties of melatonin and selenium contributed to the protective effects of MT-Se against arsenic damage. In abiotic stress, the enzyme AOX gets activated and catalyzes the oxidation of ascorbic acid, resulting in the formation of dehydroascorbic acid and water. This enzymatic activity serves to protect plant cells and maintain their survival under stressful conditions. In the current study, an increase of AOX content was observed. these findings are similar to the studies of (Ghosh et al. 2017), An increase in AOX activity was observed in wheat in response to arsenate exposure (Supplementary Fig. 2 ). Concerning proline levels, the results indicated that arsenic stress significantly elevated proline content in the leaves of both cultivars. However, T5V1 exhibited a notable recovery in proline content following the application of Cur-NPs. All the other treatments also showed an increase in proline content. Our results are similar to the findings of Anas et al. ( 2024a , b , c ). Free radicals may be scavenged by proline, which is also involved in preventing oxidative damage to cellular structures and different enzymes, maintaining the three-dimensional structure of membranes and proteins, and shielding subcellular organelles like mitochondria and chloroplasts (Zaki et al. 2023 ). The elevated proline levels observed in Cur-NPs-treated plants underscore its role in osmoprotection and membrane stabilization. This aligns with the findings of Ahmad et al. ( 2020 ) where proline accumulation was associated with improved abiotic stress tolerance. Therefore, the application of Cur-NPs aids in mitigating heavy metal toxicity. An increase in proline content under As stress was also reported by Ahmad et al. ( 2021 ). Soil-applied treatments, particularly T6 (100 mg/L Cur-NPs with arsenic), produced the most consistent improvements across morphological and biochemical parameters, including root traits, antioxidant enzyme activity, and grain yield. In contrast, foliar applications (T1 and T2) under non-stressed conditions supported growth and chlorophyll content but were less effective in limiting arsenic uptake or modulating stress markers when applied alone. The stronger response from soil-based treatments likely reflects direct interaction between nanoparticles and the root-soil interface, where arsenic mobility and ion exchange dynamics are most active. These distinctions in performance suggest that soil delivery of Cur-NPs may be more suitable for remediation protocols in contaminated environments, consistent with Yang and Yan ( 2025 ), whereas foliar applications could serve as complementary treatments under milder stress or for nutritional priming. The interaction between Cur-NPs and arsenic results in alterations in photosynthetic pigments, with chlorophyll a, chlorophyll b, total chlorophyll, and carotenoid content showing changes compared to the control group (Table 4 ). In contrast, when compared to plants treated with arsenic alone, curcumin treatment led to an increase in chlorophyll a, chlorophyll b, total chlorophyll, and carotenoid levels. This rise in total chlorophyll content may be due to curcumin's ability to interact with DNA and temporarily enhance the expression of enzymes involved in the C5 pathway for chlorophyll synthesis (Zaki et al. 2023 ). These findings align with the studies (Upadhyaya et al. 2014a , b ) indicating that arsenic reduces growth and a simultaneous decrease in photosynthetic pigments in mung bean seedlings. Cur-NPs help to mitigate the negative effects of arsenic on these seedlings. Our findings align with studies (Banerjee et al. 2023a , b ) who reported that As affects Chl a , Chl b , total Chl, and carotenoid content in 7 DAS seedlings and 28 DAS plants treated with As. However, zinc oxide nanoparticles (ZnO-NPs) enhance pigment content by preventing arsenic entry and promoting the uptake of essential nutrients such as (Ca, Mg, k and S), which are necessary for pigment biosynthesis and photosynthesis. Due to strong affinity of arsenic for–SH groups, it readily binds to proteins, thereby impacting their structure and function (Katiyar et al. 2020 ). When a plant is treated only with arsenic, it often shows a decrease in protein content due to arsenic's toxic effects on cellular metabolism (Abbas et al. 2018 ). Arsenic exposure can inhibit protein synthesis by disrupting the activity of key enzymes, interfering with nitrogen uptake, and causing oxidative stress, which damages cellular components (Tam and Wang 2020 ). Treatments like T1, T2, T3, and T4 exhibited a significant increase in protein content, while T5 and T6 also showed elevated protein levels under the influence of arsenic and Cur-NPs, in comparison to the control (C3). In contrast, protein content in C3 was reduced, likely due to the stress conditions imposed. Similar results were also reported under metal stresses (Bhat et al. 2022 ) Arsenic stress in soybean plants resulted in a decrease in protein content, while treatment with melatonin and ZnO nanoparticles improved protein levels by alleviating arsenic-induced toxicity. A similar increase in protein content was reported in a previous study by Tripathi et al. ( 2016 ). Phenolic and flavonoid compounds can act as antioxidants, the antioxidant capacity of phenolic compounds is also attributed to their ability to chelate metal ions involved in the production of free radicals, since hydroxyl groups donate hydrogen and can react with reactive oxygen species in a termination reaction, which breaks the cycle of generating new radicals (González-Moscoso et al. 2019 ). Under arsenic treatment, both flavonoid content (TFC) and phenol content (TPC) showed an increase in treatments T6 and T5 (Table 4 ). Furthermore, the application of Cur-NPs was found to further enhance the levels of TFC and TPC. This indicates that nanoparticles may play a role in amplifying the plant's antioxidant response under arsenic stress, potentially aiding in the mitigation of arsenic-induced oxidative damage through increased production of flavonoids and phenols. Our results align with the findings of González-Moscoso et al. ( 2022 ) where both TPC and TFC increase under arsenic stress, however, application of SiO 2 NPs further enhances the TPC and TFC in tomato plants. The current study found that arsenic (As) exposure, significantly reduced starch content in plants, highlighting the adverse effect of arsenic stress on starch accumulation. However, the co-application of Cur-NPs in treatments T5 and T6 markedly improved starch levels in plants subjected to arsenic, suggesting that Cur-NPs can mitigate arsenic stress and promote starch synthesis under these conditions. In contrast, starch content increased significantly in the non-arsenic treatments (T1, T2, T3, T4). These findings showed that Cur-NPs can counteract the adverse effects of arsenic on starch accumulation, offering a potential strategy for enhancing plant resilience to arsenic toxicity. Our results are similar with the findings of Bhat et al. ( 2022 ) The starch content in soybeans is adversely affected by elevated arsenic concentrations. However, the application of melatonin or zinc oxide (ZnO) nanoparticles mitigates arsenic-induced toxicity. Soybeans treated with a combination of melatonin and ZnO nanoparticles exhibited significantly higher starch levels compared to those exposed to arsenic alone. Sugars are crucial for plant growth and development as they regulate carbon and energy metabolism. Additionally, they function as osmo protecta (Bhadwal and Sharma 2022 ).nts, scavenging toxic reactive oxygen species (ROS) and offering membrane protection under abiotic stress In current findings, the sugar content was deceased under arsenic stress in C3, whereas in T5 and T6 total sugar content was increased significantly when treated with As and Cur-NPs consistent with the studies of Bano et al. ( 2022 ). According to this, total soluble carbohydrates (TSC) including glucose, fructose, and sucrose accumulation in the leaves of Brassica napus , act as a defence mechanism in response to arsenic (As). The application of Cur-NPs showed a significant reduction in arsenic uptake across all treatments, highlighting their efficacy in mitigating As stress. The result of the current findings indicates the potential of Cur-NPs in limiting arsenic translocation to aerial parts (Fig. 6 ), thereby reducing its phytotoxic effects on photosynthetic tissues. The observed reduction in arsenic uptake in Cur-NPs-treated wheat may be attributed to several interacting mechanisms. Curcumin possesses phenolic hydroxyl and diketone groups capable of forming stable complexes with metal ions, potentially immobilizing arsenic in the rhizosphere and limiting its translocation to aerial tissues, consistent with Prasad et al. ( 2021a , b ). Moreover, Cur-NPs may reduce arsenic bioavailability by adsorbing arsenic onto their surfaces or altering soil pH and redox conditions, thus affecting arsenic speciation. Another plausible mechanism involves competitive uptake inhibition, where Cur-NPs interfere with arsenic transporters or facilitate preferential absorption of essential ions, thereby limiting arsenic entry through root channels. These effects collectively restrict arsenic mobility within plant tissues, mitigating its phytotoxic impact. Similar mechanisms have been observed in other nanomaterials used for heavy metal remediation, as reported by Marslin et al. ( 2018 ), Sysak et al. ( 2023 ). Similarly, in roots, arsenic mitigation exhibited greater variability among treatments. There results align with the findings of Khan et al. ( 2024 ) where In Vigna radiata L. (mung bean), arsenic exposure led to a marked decline in germination, likely due to impaired moisture absorption, elevated lipid peroxidation, and the emergence of various chromosomal abnormalities. These effects collectively contributed to reduced seed viability. Treatment with curcumin mitigated the toxic influence of arsenic, as reflected in improved cytological integrity, biochemical balance, and antioxidant defense in the seedlings. These findings are consistent with Ghandali et al. ( 2024 ) indicating that nanoparticles can mitigate heavy metal uptake by altering the bioavailability of toxic elements through surface interactions, chelation, or soil immobilization. The results of this study have meaningful implications for arsenic-contaminated agricultural regions, particularly across South Asia, where arsenic in groundwater and soil poses a serious threat to food safety and crop productivity (Irunde et al. 2022 ). The use of green-synthesized curcumin nanoparticles offers a low-cost, biodegradable, and environmentally safe alternative to conventional soil remediation techniques. Their application improved growth and biochemical resilience in wheat under arsenic stress, suggesting Cur-NPs could be deployed in field settings through scalable and farmer-friendly methods. This approach aligns with sustainable agriculture goals and holds promise for resource-limited settings facing chronic heavy metal exposure. Compared to widely used nanoparticles such as zinc oxide (ZnO), titanium dioxide (TiO 2 ), and selenium (Se)-based formulations, Cur-NPs showed either stronger or comparable effects in managing metal-induced stress, while also aligning with eco-friendly practices. For example, foliar application of Se-NPs led to a 56% reduction in cadmium content in rice grains and supported micronutrient balance and protein levels, but control over cadmium bioavailability during digestion remained inconsistent (Hussain et al. 2024 ). TiO 2 -NPs lowered cadmium toxicity in coriander and supported pigment and proline content, though effective results required higher application rates (80–160 mg/L), and little change was noted in yield parameters under metal exposure (Sardar et al. 2022 ). ZnO-NPs applied to wheat reduced arsenic and chromium toxicity and supported moderate antioxidant activity (Iqbal et al. 2024 ), yet the increase in grain yield (26–41%) was still less than that observed in the current study. In contrast, Cur-NPs at 100 mg/L lowered arsenic uptake by over 70%, supported pigment recovery by nearly 80%, and raised grain yield by over 140%. These findings suggest that Cur-NPs provide a more responsive and efficient strategy for managing metal stress at lower doses, with the added benefit of being plant-derived and biodegradable. Recent research has explored carbon-based nanomaterials for heavy metal detoxification, with promising outcomes. For instance, graphitic carbon nitride (C 3 N 4 ) nanosheets significantly alleviated arsenic-induced phytotoxicity in rice, decreasing arsenic accumulation in roots and shoots by 25% and 36%, respectively (Ma et al. 2021 ). In addition to reducing metal uptake, C 3 N 4 modulated the expression of key transporters like OsNIP1;1 and OsPT4, limiting translocation and internal toxicity. Unlike metal-based formulations, nanocarbon materials such as C 3 N 4 exert minimal disturbance on macronutrient balance, maintaining plant physiological integrity. While Cur-NPs similarly restricted arsenic uptake and improved plant health, their added bio-derived antioxidative and chelating functionality positions them as more biologically integrated solutions compared to synthetic graphitic analogs. Moreover, the unique functionality of curcumin lies in its dual role as both an antioxidant and a metal-chelating compound, which collectively underpins the superior performance of Cur-NPs under arsenic stress. Curcumin’s molecular structure—rich in phenolic hydroxyl and β-diketone groups—enables direct complexation with metal ions such as arsenic, reducing their bioavailability and systemic translocation (Prasad et al. 2021a , b ). Simultaneously, these functional moieties participate in ROS scavenging through hydrogen donation and electron transfer reactions, thereby modulating oxidative damage within cellular compartments (Sarkar et al. 2019 ; Khan et al. 2022 ). In contrast, conventional metal-based nanoparticles such as ZnO (Wang et al. 2023 ), TiO 2 (Babaei et al. 2025 ), and Se-NPs (Qi et al. 2021 ) typically exert protective effects through singular mechanisms—either by influencing the redox signalling pathway or by adsorbing ions at the root-soil interface (Ahmed et al. 2020 ). The combinatorial antioxidative and chelating action of Cur-NPs facilitates both physiological stability and detoxification, rendering them more effective in mitigating arsenic-induced stress in plants compared to traditional formulations. The findings revealed consistent genotypic differences between Barani-70 and NARC-09 in their responses to Cur-NP applications under arsenic stress. Barani-70 generally performed better in terms of physiological, morphological, and biochemical traits—including biomass production, protein accumulation, and antioxidant enzyme regulation. For example, treatments such as T1V1, T5V1, and T6V1 showed stronger growth recovery and stress mitigation compared to their NARC-09 counterparts (T1V2, T5V2, and T6V2). This suggests that Barani-70 may possess intrinsic mechanisms, such as improved nanoparticle interaction or more efficient oxidative stress responses, that confer a greater tolerance to arsenic stress when treated with Cur-NPs. These findings underscore the importance of genotype-specific compatibility in nanoparticle-assisted remediation strategies and suggest that cultivar selection could significantly influence the field effectiveness of nanotechnology-based interventions. While this study was conducted under controlled conditions, future validation across diverse agroecological zones remains essential to evaluate consistency in Cur-NPs performance. Arsenic retention, bioavailability, and nanoparticle behaviour vary with soil physicochemical characteristics, necessitating multi-location field trials on varied soil textures and mineral profiles. In parallel, transcriptomic or gene expression analyses can provide molecular insights into regulatory pathways modulated by Cur-NPs, particularly genes related to metal transport, ROS detoxification, and chlorophyll biosynthesis. Such analyses could reveal cultivar-specific responses, aiding in the development of genetically informed remediation strategies. Additionally, standardization of Cur-NP formulations—considering optimal concentration, particle size, and delivery method—will be crucial for transitioning this approach from laboratory to scalable field application. These aspects will contribute to designing precise, safe, and economically viable nanoparticle deployment protocols tailored for arsenic-affected agricultural systems. Conclusion This study investigated the efficacy of green-synthesized curcumin nanoparticles (Cur-NPs) in mitigating arsenic-induced stress in wheat cultivars Barani-70 and NARC-09, focusing on physiological, biochemical, and agronomic responses. The results demonstrated that Cur-NPs effectively reduced arsenic uptake in both roots and leaves, with the greatest reductions observed in T5 and T6 treatments. Enhanced starch, protein, and sugar content, along with improved phenolic and flavonoid levels, highlight the protective role of Cur-NPs against arsenic-induced metabolic disruptions. Moreover, Cur-NPs significantly alleviated oxidative stress by modulating the activity of antioxidant enzymes such as SOD, POD, and CAT, and reducing key stress markers, including MDA and H 2 O 2 levels. Improved proline accumulation and chlorophyll content further underscored the ability of Cur-NPs to enhance plant resilience under arsenic stress. Agronomic traits such as plant height, biomass yield, and grain yield also showed marked improvements with Cur-NPs treatments, indicating their potential to enhance crop productivity in arsenic-contaminated soils. These findings present Cur-NPs as a viable nanotechnological approach for sustainable agriculture, reducing heavy metal toxicity and improving crop performance. Although total arsenic content was quantified via atomic absorption spectroscopy, this study did not differentiate between trivalent and pentavalent arsenic species. Incorporating arsenic speciation in future analyses could help elucidate the chemical forms retained or transformed in plant tissues under Cur-NP treatment, offering more mechanistic insight into Cur-NP-mediated detoxification. Future research should focus on scaling up the application of Cur-NPs to field conditions and assessing their long-term environmental implications to optimize their use in arsenic-affected agricultural systems. Supplementary Information Below is the link to the electronic supplementary material. Supplementary file1 (DOCX 74115 kb) (72.4MB, docx) Author contributions Nimra Tahir: Performed experiment, Conceptualization, revision, and writing-editing. Minhas Elahi: Acquisition of data, data interpretation, writing-editing, and statistical analysis. Rimsha Aslam: Acquisition of data. Prof. Dr. Umar Masood Quraishi: Conceptualization, acquisition of funds, data interpretation and revision. Funding The research leading to these results received funding from Quaid-i-Azam University for 2024. Data availability Data will be made available on request. Declarations Conflict of interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Footnotes Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. 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