The synthesized zinc oxide/silicon dioxide@graphene oxidenanocomposite (ZnO/SiO2@GO) enhances quinoa tolerance to high salinity

The synthesized zinc oxide/silicon dioxide@graphene oxidenanocomposite (ZnO/SiO2@GO) enhances quinoa tolerance to high salinity | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The synthesized zinc oxide/silicon dioxide@graphene oxidenanocomposite (ZnO/SiO 2 @GO) enhances quinoa tolerance to high salinity Enas G. Badran, Nouriya S. Mohammed, Mamdouh M. Nemat Alla, Mohamed M. EL-Zahed This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6165086/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract The nanocomposite zinc oxide/silicon dioxide@graphene oxide (ZnO/SiO 2 @GO) was synthesized by the decoration of ZnO nanoparticles (NPs) and SiO 2 NPs onto GO. The synthesized nanocomposite was used as seed soaking for modulation of tolerance of quinoa ( Chenopodium quinoa ) to high salinity stress. Seeds were soaked in 40 mg ZnO/SiO 2 @GO L − 1 of the nanocomposite for 6 h thereafter NaCl was applied at 200 mM, 400 mM and 500 mM -on the 10th day of sowing- for a duration of 21 days. The results demonstrated that high NaCl (400 mM and 500 mM) significantly reduced growth parameters, photosynthetic pigments, protein, insoluble and total sugars, phenolics and activities of peroxidase and catalase. Conversely, soluble sugars, lipid peroxide (malondialdehyde, MDA) and hydrogen peroxide (H 2 O 2 ) levels were elevated, with the effects being more pronounced at higher NaCl concentrations. However, using ZnO/SiO 2 @GO as a seed priming agent effectively mitigated the adverse effects of NaCl on these parameters. Growth was enhanced, while reductions in pigments, protein, sugars, phenolics, and enzymatic activities were counteracted. Additionally, elevated levels of soluble sugars, MDA, and H 2 O 2 were suppressed, approaching control levels. These findings highlight the severe impacts of high salinity stress and indicate that treatment with the ZnO/SiO2@GO nanocomposite can mitigate these adverse effects. Therefore, this nanocomposite could be considered a safe and efficient protectant to enhance quinoa's tolerance to high NaCl levels. catalase growth peroxidase phenolics salinity nanocomposite Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Highlights • The nanocomposite ZnO/SiO@GO was synthesized • High NaCl levels reduced quinoa’s growth, pigments, protein, sugars, phenolics and peroxidase and catalase activities but increased soluble sugars, MDA and HO • ZnO/SiO@GO mitigated the detrimental effects of NaCl • The nanocomposite enhances quinoa’s tolerance and is recommended as a safe protectant against high NaCl levels 1 Introduction Quinoa ( Chenopodium quinoa ) has emerged as an ideal food security crop due to its exceptional nutritional profile. It is considered an alternative to cereals, providing comparable energy levels to wheat, rice, beans, and corn while also helping regulate blood sugar levels and prevent degenerative diseases [ 1 ]. Additionally, quinoa is rich in proteins, minerals, vitamins, high-quality fatty acids, and various secondary metabolites [ 2 ]. Despite its beneficial characteristics, quinoa is regarded as a facultative halophyte, with a salt tolerance greater than wheat, corn, and barley, tolerating up to 250 mM NaCl in most genotypes [ 3 ]. However, high salinity disrupts several vital physiological and biochemical processes in plants [ 4 ], leading to osmotic stress, water deficiency, ion toxicity and imbalance, membrane damage, metabolic dysfunction, enzyme inhibition, alterations in growth regulators, and the production of reactive oxygen species (ROS) [ 5 , 6 , 7 ]. ROS can damage cellular membranes, nucleic acids, proteins, and carbohydrates [ 8 ], but their toxic effects are counteracted by enzymatic and non-enzymatic antioxidative defense systems [ 9 ]. Salinity tolerance in plants is influenced by stress adaptation effectors [ 10 ]. Although quinoa exhibits higher salinity tolerance than other cereals, exposure to high NaCl concentrations can still have detrimental effects. While plants possess endogenous mechanisms to cope with salinity stress, external interventions are often necessary [ 6 , 10 ]. Given quinoa's nutritional value, various strategies, including the application of nanoparticles (NPs), have been explored to enhance its salinity tolerance, facilitating cultivation in saline soils. Research indicates that NPs can maintain crucial biochemical and physiological functions in plants, as demonstrated in cotton [ 11 ] and basil [ 12 ]. Zinc (Zn) and silicon (Si) play essential roles in plant physiology. Zn is involved in enzyme activation, ion homeostasis, and overall plant growth, development, and yield [ 13 ]. Si enhances growth, yield, crop quality, photosynthesis, nitrogen fixation, and stress tolerance [ 14 ]. Furthermore, graphene oxide has been found to improve chlorophyll content, relative water content, and growth performance in plants such as Silybum marianum [ 15 ]. This study aimed to synthesize a high-performance nanocomposite (ZnO/SiO2@GO) by decorating ZnO and SiO2 NPs onto GO and evaluate its effectiveness in mitigating the detrimental effects of high salt stress on quinoa. Seeds primed with ZnO/SiO2@GO were cultivated, and the resulting seedlings were exposed to elevated NaCl levels. The impacts on growth parameters, pigments, sugars, proteins, MDA, H 2 O 2 , phenolics, and peroxidase and catalase activities were assessed to determine whether the nanocomposite could enhance quinoa's tolerance to salinity, positioning it as a promising protective agent against high salt stress. 2 Materials and Methods 2.1 Synthesis of zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO 2 @GO) ZnO was synthesized by gradual addition of KOH solution into 0.2 M Zn(NO 3 ) 2 solution while being vigorously stirred and centrifuged at 5000 ×g for 15 min. The resulting ZnO NPs were calcined at 500°C for 3 h [ 16 ].SiO 2 was synthesized by dropwise addition of Na 2 SiO 3 solution to 33% ammonia/absolute ethanol (3:1 v/v) and centrifuged at 5000 ×g for 15 min. SiO 2 NPs were dried at 50°C for 24 h and 185°C for additional 5 h [ 17 ]. GO was synthesized from the oxidation of graphite [ 18 ] using H 2 SO 4 /HNO 3 (3:1 v/v) in ice-water bath with agitation for 3 hat 35°C then distilled water was gradually added while being vigorously stirred. After changing the color into dark brown, the reaction mixture was diluted then H 2 O 2 was added dropwise. The resultant GO was adjusted to pH 7and centrifuged for 15 min then GO was dried at 80°C. Decoration of ZnO NPs and SiO2 NPs onto GO was performed by dispersing GO in chitosan/polyvinyl alcohol (CS/PVA) [ 19 ] previously prepared from dissolving PVA in 5% acetic acid followed by the addition of CS [ 20 ]. CS/PVA containing GO was ultrasonicated for an hour at 25°C. ZnO NPs and SiO 2 NPs were added and stirred for an hour, and then glutaraldehyde was added to the stirring suspension. 2.2 Plant material and growth conditions Certified seeds of quinoa [ Chenopodium quinoa (L.)] were obtained from Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt. Seeds were surface sterilized in 1% w/v sodium hypochlorite for 5 min, rinsed with water several times and soaked for 6 h either in water or in ZnO/SiO 2 @GO (40 mg L − 1 ). Then the seeds of both groups were planted in plastic pots (40 × 25 × 15 cm), each contained 3 kg of soil (clay: sand; 2:1 w/w), approximately 2 cm-depth and spaced 5 cm apart in rows and allowed to grow at 22 ± 2/12 ± 2°C, day/night temperature with a 12-h photoperiod at 420–460 µmol m − 2 s − 1 PPFD. All pots received water (200 ml per pot) daily for 3 days followed by Long Ashton nutrient solution up to the 10th day, then pots of both groups were divided into four subgroups for treatment with NaCl at 0 mM (control), 200 mM, 400 mM and 500 mM every 3 days (200 ml per pot) for the following 21 days. When seedlings were 35-day-old, samples were collected, washed thoroughly with water, plotted dry between layers of tissue, and separated into shoots and roots. Shoot height, root length and fresh weights were determined, and then dried at 80°C for 2 days for dry weight determination from which water contents were calculated. The new leaves and the upper parts were frozen in liquid N and used for analyses. 2.3 Determination of photosynthetic pigments, protein and soluble sugar Chlorophylls (a and b) and carotenoids in fresh tissues were extracted with 80% acetone, then absorbance was read at 663.2, 646.8 and 470 nm. Pigment contents were calculated as µg ml − 1 then converted to µg unit weight − 1 [ 21 ]. Protein in fresh tissue was extracted with chilled acetone, centrifuged and the residues were allowed to dry at room temperature to obtain acetone powders [ 22 ]. Aliquots of the acetone powders were mixed with Tris-HCl (0.05 M, pH 9.0) for 10 min at 4°C and then centrifuged at 20000 ×g for 15 min at 4°C. Protein content was determined spectrophotometrically at 595 nm by reaction with Coomassie Brilliant Blue G-250 [ 23 ].Soluble sugars were extracted in 80% ethanol [ 24 ]. An aliquot was mixed with 3 ml anthrone reagent (8.6 mM anthrone in 80% H 2 SO 4 ) and heated for 10 min, then cooled in ice bath for 30 min and absorbance was read at 623 nm [ 25 ]. The residues remained from extraction of soluble sugars were digested in 1.6 M perchloric acid for the determination of non-soluble sugars using anthrone. 2.4 Determination of lipid peroxidation (malondialdehyde, MDA), hydrogen peroxide (H 2 O 2 ) and phenolic compounds MDA in fresh tissues was extracted with 10% Trichloroacetic acid (TCA, w/v) then centrifuged at 12000 ×g for 15 min and absorbance was read at 450, 532, and 600 nm after reaction with tribromo-arsenazo colorimetry using 10% TCA containing 0.5% thiobarbituric acid from the equation: 6.45(A532 − A600) − 0.56(A450) [ 26 ]. H 2 O 2 was extracted with 0.1% TCA and centrifuged at 12000 ×g for 15 min at 4°C, assayed in potassium phosphate buffer (10 mM, pH 7.0) containing 1 M KI and absorbance was measured at 390 nm [ 27 ]. Phenolic compounds were determined using Folin–Ciocalteu reagent and absorbance was read at 765 nm [ 28 ]. 2.5 Extraction and assay of peroxidase and catalase Peroxidase and catalase in fresh tissues were extracted with 50 mM sodium phosphate buffer (pH 7.0) containing 1 mM EDTANa 2 and 0.5% Polyvinylpolypyrrolidone under cold conditions and centrifuged at 13000 ×g for 30 min at 4°C. Peroxidase activity was assayed following the ascorbate oxidation by the decrement in absorption at 290 nm [ 29 ]. Catalase activity was assayed by controlling the H 2 O 2 absorbance reduction at 240 nm [ 30 ]. 2.6 Statistical analysis The experiment was repeated twice and samples for each analysis were taken in triplicates from both experiments (n = 6) and mean values (± SD) were applied. The experiment was designed as a complete randomized block consisting of 160 pots (2 sets for ZnO/SiO 2 @GO treatments: control, 40 mg L − 1 ) x (4 sets for NaCl treatments: control, 200 mM, 400 mM and 500 mM) x (10 replications) x (2 repetitions). The full data were subjected to analysis of variance (ANOVA) followed thereafter by least significant differences (LSD) at 5% level. 3 Results Figure 1 shows that NaCl concentrations of 400 mM and 500 mM significantly reduced the shoot height and root length of 35-day-old quinoa seedlings compared to control values. However, the reduction caused by 200 mM NaCl was not significant. The extent of the decrease increased with higher NaCl concentrations. Similar significant reductions were observed in fresh and dry weights as well as in the water content of both shoots and roots, with all NaCl concentrations affecting these parameters. Overall, the reductions in shoot height, root length, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, shoot water content, and root water content ranged from 18–56%, 10–36%, 34–68%, 38–79%, 16–40%, 27–66%, 4–12%, and 3–11%, respectively. However, pre-soaking quinoa seeds in the synthesized ZnO/SiO 2 @GO nanocomposite (40 mg L − 1 ) improved the growth parameters under normal conditions, resulting in increases of approximately 17%, 11–13%, and 5–11% in shoot and root length, fresh weight, and dry weight, respectively, along with slight increases in water content. Furthermore, the nanocomposite completely mitigated the negative effects of all NaCl concentrations on these growth parameters, making NaCl have no significant effect except for the 500 mM concentration, which still had a significant effect on shoot fresh weight. The highest decreases induced by NaCl in the presence of the nanocomposite were limited to no more than 13%, 11%, 6%, and 1% in shoot and root length, fresh weight, dry weight, and water content, respectively, compared to the control. In contrast, without the nanocomposite, the reductions reached 56%, 79%, 66%, and 12%, respectively. Figure 2 shows that NaCl treatment significantly reduced the contents of chlorophylls and carotenoids in quinoa compared to untreated controls. The decreases induced by 200 mM NaCl were non-significant, but they increased as NaCl concentration rose. The reductions in chlorophyll a, chlorophyll b, and carotenoids at 400 mM NaCl were approximately 38%, 39%, and 35%, respectively, compared to the untreated control, while 500 mM NaCl caused reductions of 50%, 40%, and 45%, respectively. Total pigments were also significantly reduced by NaCl, with high concentrations leading to reductions of 37–46%. The application of ZnO/SiO 2 @GO under normal conditions resulted in increases of approximately 8–11% in chlorophyll and carotenoid content relative to the control. In NaCl-treated samples, the nanocomposite greatly mitigated these decreases, bringing pigment levels closer to those of the untreated control. In fact, some increases were observed, particularly at 200 mM and 400 mM NaCl (2–8%). In Fig. 3 , protein content was significantly decreased by 400 mM and 500 mM NaCl, with the decrease being about 35% and 46%, respectively, compared to the untreated control. The decrease induced by 200 mM NaCl was insignificant. Conversely, NaCl treatment caused significant increases in soluble sugars, with increases ranging from 22–24% for all concentrations. Insoluble sugar content was significantly reduced by 18%-48%, with the highest reduction observed at 500 mM NaCl. Consequently, total sugar content decreased under all NaCl treatments, with the greatest reduction (27%) occurring at 500 mM, compared to 16% and 6% reductions at 200 mM and 400 mM, respectively. However, pre-soaking quinoa seeds in ZnO/SiO₂@GO nanocomposite as a seed pretreatment greatly increased protein content by about 21% in the absence of salinity and counteracted the effects of NaCl. Additional increases in protein content of 15%, 12%, and no change were observed in response to 200 mM, 400 mM, and 500 mM NaCl, respectively. Similarly, the nanocomposite mitigated the increases in soluble sugars, returning them to levels near the control, while the decreases in insoluble and total sugars were largely compensated, with reductions no greater than 5%. As shown in Fig. 4 , MDA and H 2 O 2 contents were significantly elevated in NaCl-treated quinoa, with higher concentrations leading to greater accumulation. The increases induced by 200 mM, 400 mM, and 500 mM NaCl were approximately 125%, 160%, and 184% for MDA, and 103%, 147%, and 159% for H 2 O 2 , respectively. In contrast, NaCl treatment caused significant decreases in the endogenous contents of phenolics, with reductions of about 29%, 42%, and 49% for 200 mM, 400 mM, and 500 mM NaCl, respectively. However, pre-soaking quinoa seeds in ZnO/SiO 2 @GO greatly suppressed the accumulation of MDA and H 2 O 2 , fully mitigating the effects of NaCl. Under normal conditions, the nanocomposite induced a 10% increase in phenolics, and it limited the decrease induced by the highest NaCl concentration to no more than 3%. In concomitant, NaCl treatment significantly inhibited the activities of peroxidase and catalase, with the effect being more pronounced at 500 mM NaCl compared to 400 mM and 200 mM (Fig. 5 ). The inhibition reached about 51 − 31% for peroxidase and 49 − 28% for catalase activities. However, seed soaking in ZnO/SiO 2 @GO led to substantial increases in both peroxidase and catalase activities in normally grown samples (about 17% and 25%, respectively). Additionally, the inhibitory effects of NaCl on peroxidase and catalase activities were largely mitigated, and further enhancements were observed, with increases of 5%-11% for peroxidase and 6%-17% for catalase compared to control values. 4 Discussion Quinoa's exceptional nutritive profile and its higher tolerance to salinity compared to other cereals make it an ideal candidate for cultivation in saline and marginal soils. The ZnO/SiO 2 @GO nanocomposite was synthesized in this study to modulate and enhance quinoa's tolerance to high salinity, providing an efficient protectant against elevated salt levels. High NaCl concentrations (400 mM and 500 mM) significantly reduced quinoa growth, photosynthetic pigments, protein, insoluble and total sugars, and phenolic compounds while inhibiting peroxidase and catalase activities. In contrast, soluble sugars, MDA, and H 2 O 2 levels were elevated under high salinity. These results confirm the negative effects of high salinity on quinoa growth, despite its higher salt tolerance than wheat, corn, and barley [ 3 ]. Generally, salinity disrupts several plant processes by altering water potential, causing ion imbalance and toxicity, impairing cell division and expansion, and generating reactive oxygen species (ROS), which further damage cellular components [ 4 , 31 ]. Salinity may reduce plant growth by causing chlorophyll degradation, as chlorophyll is a critical determinant for photosynthesis. This study supports findings by Mazumder et al. [ 32 ], who observed that salt stress led to pigment degradation due to the vulnerability of photosystems. Salinity can also disrupt electron transport in the photosynthetic system, leading to oxidative damage. Carotenoids, however, protect chlorophyll from photooxidation by removing oxygen from excited chlorophyll-oxygen complexes. According to Nemat Alla et al. [ 33 ], the inhibition of carotenoids in maize seedlings resulted in chlorophyll degradation, which compromised the photosynthetic capacity. Similarly, in this study, NaCl treatment reduced carotenoid content, potentially exacerbating the chlorophyll degradation and negatively impacting photosynthesis. However, NaCl increased soluble sugars to act as osmolytes against salinity for membrane protection and osmotic adjustment. Salinity increased soluble sugars, which act as osmolytes to protect membranes and facilitate osmotic adjustment to survive extreme osmotic stress. These sugars might have resulted from the hydrolysis of polysaccharides rather than new synthesis. Elevated soluble sugars provide protection against salinity by stabilizing proteins and cell membranes under osmotic stress [ 34 ]. Conversely, protein synthesis appeared to be impaired by the salt-induced decrease in photosynthetic activity. This is supported by the alterations in peroxidase and catalase activities, which are vital antioxidant enzymes that protect against reactive oxygen species (ROS) generated under stress which are destructive to proteins and DNA. The substantial increase in MDA and H 2 O 2 levels under NaCl treatment, coupled with inhibited antioxidant enzyme activity, suggests the induction of oxidative stress. Under oxidative stress, ROS damage proteins, DNA, and lipids, further impairing growth and metabolism. Peroxidase and catalase can scavenge H 2 O 2 [ 31 ]. The inhibition of peroxidase and catalase activities under salt stress impedes ROS detoxification, exacerbating cellular damage and limiting growth. The present work declares that quinoa tolerated low concentrations of NaCl but suffers from the high concentration which resulted in deteriorations. However, soaking quinoa seeds in the ZnO/SiO 2 @GO nanocomposite prior to salinity treatment improved growth and mitigated the adverse effects of NaCl. The nanocomposite enhanced antioxidant activities, alleviated oxidative stress, and improved pigment content, protein synthesis, and photosynthetic capacity. This suggests that ZnO/SiO 2 @GO plays a key role in enhancing quinoa's tolerance to high salinity. The inclusion of Zn, SiO 2 , and graphene oxide (GO) in the nanocomposite likely contributed to improved plant health. Zn supports enzymatic functions and ion homeostasis [ 13 ], SiO 2 provides stress tolerance and boosts photosynthesis [ 14 ], and GO improves chlorophyll content and water retention [ 15 ]. Nanoparticles (NPs) have been reported to increase plant stress tolerance by preserving critical physiological and biochemical functions Kalteh et al. [ 12 ] found that NPs increased growth, yield, and starch content in wheat subjected to drought stress. In consistent with the present results, NPs mitigated abiotic stresses and improved wheat growth under stress conditions [ 35 ]. Also, NPs increased plant tolerance and improved physiological processes of flaxseed under stress [ 36 ]. In this study, the nanocomposite ameliorated quinoa growth and alleviated the reduction in chlorophyll and carotenoid levels caused by NaCl, possibly by reducing oxidative stress and enhancing the synthesis of these pigments. The nanocomposite also mitigated the accumulation of MDA and H 2 O 2 , and increased the activity of antioxidant enzymes, highlighting its role in protecting quinoa from oxidative damage under saline conditions. These improvements in photosynthetic capacity, combined with elevated pigment content, total sugars, and protein levels, point to enhanced plant growth under stress. Zhan et al. [ 35 ] who concluded that NPs promoted wheat growth by elevating chlorophyll content. Mehrian et al. [ 37 ] indicated that NPs significantly elevated redox enzyme activity in Lycopersicon esculentum . Similarly, Saha and Gupta [ 38 ] reported that Ag NPs enhanced in vitro Swertia chirata growth by the activation of antioxidants for scavenging free radicals as a defense mechanism in response to abiotic stress [ 39 ]. So, the amelioration in quinoa growth by the nanocomposite under high salt stress synchronized with elevated photosynthetic pigments, total sugars and protein contents could reveal an improvement of photosynthetic incapacity. Meanwhile, the nanocomposite suppressed the accumulated MDA and H 2 O 2 concomitantly with enhancing phenolic compounds and the activities of peroxidase and catalase pointing at ameliorated antioxidant performance that would result in modulating quinoa tolerance to high salinity and improving growth. 5 Conclusions The high concentrations of NaCl significantly impacted quinoa growth, pigments, protein, and sugar content, and induced oxidative stress. MDA and H 2 O 2 levels increased, while phenolic compounds and antioxidant enzyme activities were inhibited. These findings clearly declare that quinoa suffered from high salinity levels. However, the application of a synthesized ZnO/SiO 2 @GO nanocomposite (40 mg L − 1 ) through seed soaking effectively improved quinoa's growth parameters, pigment content, and protein synthesis, demonstrating its potential to alleviate the harmful effects of high salinity. The nanocomposite seemed beneficial in eliminating the oxidative stress by suppressing MDA and H 2 O 2 levels and enhancing phenolic compounds and antioxidant enzyme activities, making it a promising strategy for improving quinoa tolerance to high salinity. These findings highlight the nanocomposite's ability to enhance quinoa's resilience under saline conditions, which is crucial for growing quinoa in saline-affected soils. Declarations Author contributions MN: planed the scientific experiments, performed data analysis, wrote, edited and reviewed the manuscript; EB, NM and ME: performed the experiments. All the authors have approved the final manuscript. Ethics approval and consent to participate The plant seeds used in this study were obtained from Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt. The plants used in the study comply with local guidelines. The plant samples collected in this study were cultivated varieties and planted in the greenhouse of the Faculty of Science, Damietta University, Egypt. Samples are allowed to be picked for scientific research purposes. Consent to Participate declaration Seeds were obtained from Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt. All the authors have approved the final manuscript. Conflict of interest The authors declare that there is no conflict of interest. Funding Open access funding is provided by The Science, Technology & Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB). Competing interests The authors declare that they have no competing interests. Consent to Publish declaration Not applicable. This work does not involve patient(s). Data availability Data availability has been declared in the ‘Declaration’ in the system. Data availability statement that was provided in the manuscript has been added on the editorial system as well. 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Molecules 25:5511 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 28 Apr, 2025 Reviews received at journal 27 Apr, 2025 Reviews received at journal 26 Apr, 2025 Reviewers agreed at journal 19 Apr, 2025 Reviewers agreed at journal 18 Apr, 2025 Reviewers invited by journal 18 Apr, 2025 Editor assigned by journal 03 Apr, 2025 Editor invited by journal 02 Apr, 2025 Submission checks completed at journal 02 Apr, 2025 First submitted to journal 02 Apr, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6165086","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":448637900,"identity":"6fd5a0a1-c38a-4b46-845d-ac11f82e65a7","order_by":0,"name":"Enas G. Badran","email":"","orcid":"","institution":"Damietta University","correspondingAuthor":false,"prefix":"","firstName":"Enas","middleName":"G.","lastName":"Badran","suffix":""},{"id":448637901,"identity":"aa43fe6e-545c-4748-ba78-83271f8c6ea1","order_by":1,"name":"Nouriya S. Mohammed","email":"","orcid":"","institution":"Sebha University","correspondingAuthor":false,"prefix":"","firstName":"Nouriya","middleName":"S.","lastName":"Mohammed","suffix":""},{"id":448637902,"identity":"48089b3d-81c7-42d5-b8ce-f3838265ebd0","order_by":2,"name":"Mamdouh M. Nemat Alla","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9ElEQVRIiWNgGAWjYFADduYGhg9sIFYCcRokGJgZGxhnkKyFmYcYLbrtp9MefMyxq+NvZmz8bFN2mIGfPceA4cMv3FrMzuRuN5y5LVlC4jBjs3TOucMMkj1vDBhn9uHRciB3mzTvNmYJhsOMDdK5bYcZDG7kGDDz9uDRcv7tNum/2+ol5IG2/LYEarEHafmLT8sNoC2M2w5LGBxmbJNmBNkiAdTC8AOflrfbJHu3HZfcCNRi2XMunUfizLOCg70N+ByWu03i57ZqfrnjzYdv/CizluNvT9744Mcf3FowAA+IOMDYRoIWKCDFllEwCkbBKBjuAAClK1Q85YYuhgAAAABJRU5ErkJggg==","orcid":"","institution":"Damietta University","correspondingAuthor":true,"prefix":"","firstName":"Mamdouh","middleName":"M. Nemat","lastName":"Alla","suffix":""},{"id":448637903,"identity":"61fa5cd7-4bf9-4e30-a7b8-c748101c02b6","order_by":3,"name":"Mohamed M. EL-Zahed","email":"","orcid":"","institution":"Damietta University","correspondingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"M.","lastName":"EL-Zahed","suffix":""}],"badges":[],"createdAt":"2025-03-05 19:08:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6165086/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6165086/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":81519907,"identity":"b21c4c0b-bf70-446f-b040-4871d1476f10","added_by":"auto","created_at":"2025-04-28 07:54:48","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":50865,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in growth parameters in shoots and roots of 35-day old quinoa subjected to NaCl treatment for the preceding 21 days, following seed soaking in the zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) at a concentration of 20 mg L\u003csup\u003e-1\u003c/sup\u003e. Black bars represent the control (without ZnO/SiO2@GO) while white bars represent treated samples (with ZnO/SiO2@GO). Values are expressed as means ± SD. Analysis of variance (ANOVA) was preformed followed by least significant differences (LSD) at the 5% level (vertical bars: solid: for control, dotted for treated). Water contents were derived from original means; thus no ANOVA or LSD was applied.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6165086/v1/482a8e989cb6e23388a91b9c.png"},{"id":81519908,"identity":"157fa6ec-f1b3-4aac-a37a-1648d3e2894d","added_by":"auto","created_at":"2025-04-28 07:54:48","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":26968,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in pigment contents in shoots of 35-day old quinoa subjected to NaCl treatment for the preceding 21 days, following seed soaking in the zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) at a concentration of 20 mg L\u003csup\u003e-1\u003c/sup\u003e. Black bars represent the control (without ZnO/SiO2@GO) while white bars represent treated samples (with ZnO/SiO2@GO). Values are expressed as means ± SD. Analysis of variance (ANOVA) was preformed followed by least significant differences (LSD) at the 5% level (vertical bars: solid: for control, dotted for treated). Total pigments were derived from original means; thus no ANOVA or LSD was applied.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6165086/v1/bb42843824e964a33ec99f31.png"},{"id":81520663,"identity":"4c3f4fb4-6976-4b84-a0ad-aa1524344628","added_by":"auto","created_at":"2025-04-28 08:02:48","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":29812,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in contents of protein and sugars in shoots of 35-day old quinoa subjected to NaCl treatment for the preceding 21 days, following seed soaking in the zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) at a concentration of 20 mg L\u003csup\u003e-1\u003c/sup\u003e. Black bars represent the control (without ZnO/SiO2@GO) while white bars represent treated samples (with ZnO/SiO2@GO). Values are expressed as means ± SD. Analysis of variance (ANOVA) was preformed followed by least significant differences (LSD) at the 5% level (vertical bars: solid: for control, dotted for treated). Total sugars were derived from original means; thus no ANOVA or LSD was applied.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6165086/v1/8cbecd9d4fe866deb9bb889f.png"},{"id":81522375,"identity":"2c14816b-703e-474c-826a-b8e53342fda9","added_by":"auto","created_at":"2025-04-28 08:10:48","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":36847,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in contents of lipid peroxide (MDA), hydrogen peroxide and phenolics in shoots of 35-day old quinoa subjected to NaCl treatment for the preceding 21 days, following seed soaking in the zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) at a concentration of 20 mg L\u003csup\u003e-1\u003c/sup\u003e. Black bars represent the control (without ZnO/SiO2@GO) while white bars represent treated samples (with ZnO/SiO2@GO). Values are expressed as means ± SD. Analysis of variance (ANOVA) was preformed followed by least significant differences (LSD) at the 5% level (vertical bars: solid: for control, dotted for treated).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6165086/v1/c5305653a6ba311bcfce4ac5.png"},{"id":81523127,"identity":"8ff91bad-92f1-4380-8527-0d8e602a6cb2","added_by":"auto","created_at":"2025-04-28 08:18:48","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":22391,"visible":true,"origin":"","legend":"\u003cp\u003eChanges in peroxidase and catalase activities in shoots of 35-day old quinoa subjected to NaCl treatment for the preceding 21 days, following seed soaking in the zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) at a concentration of 20 mg L\u003csup\u003e-1\u003c/sup\u003e. Black bars represent the control (without ZnO/SiO2@GO) while white bars represent treated samples (with ZnO/SiO2@GO). Values are expressed as means ± SD. Analysis of variance (ANOVA) was preformed followed by least significant differences (LSD) at the 5% level (vertical bars: solid: for control, dotted for treated).\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6165086/v1/77afa9f33a0659c776b3b197.png"},{"id":81524349,"identity":"5a0013ba-32ac-4e1a-8d0c-33e051fdcdf1","added_by":"auto","created_at":"2025-04-28 08:26:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":821479,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6165086/v1/3ad373a9-ec86-464f-9507-f1a430a4aac6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eThe synthesized zinc oxide/silicon dioxide@graphene oxidenanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) enhances quinoa tolerance to high salinity\u003c/p\u003e","fulltext":[{"header":"Highlights","content":"\u003cp\u003e\u0026bull; The nanocomposite ZnO/SiO@GO was synthesized\u003c/p\u003e\u003cp\u003e\u0026bull; High NaCl levels reduced quinoa\u0026rsquo;s growth, pigments, protein, sugars, phenolics and peroxidase and catalase activities but increased soluble sugars, MDA and HO\u003c/p\u003e\u003cp\u003e\u0026bull; ZnO/SiO@GO mitigated the detrimental effects of NaCl\u003c/p\u003e\u003cp\u003e\u0026bull; The nanocomposite enhances quinoa\u0026rsquo;s tolerance and is recommended as a safe protectant against high NaCl levels\u003c/p\u003e"},{"header":"1 Introduction","content":"\u003cp\u003eQuinoa (\u003cem\u003eChenopodium quinoa\u003c/em\u003e) has emerged as an ideal food security crop due to its exceptional nutritional profile. It is considered an alternative to cereals, providing comparable energy levels to wheat, rice, beans, and corn while also helping regulate blood sugar levels and prevent degenerative diseases [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Additionally, quinoa is rich in proteins, minerals, vitamins, high-quality fatty acids, and various secondary metabolites [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDespite its beneficial characteristics, quinoa is regarded as a facultative halophyte, with a salt tolerance greater than wheat, corn, and barley, tolerating up to 250 mM NaCl in most genotypes [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. However, high salinity disrupts several vital physiological and biochemical processes in plants [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], leading to osmotic stress, water deficiency, ion toxicity and imbalance, membrane damage, metabolic dysfunction, enzyme inhibition, alterations in growth regulators, and the production of reactive oxygen species (ROS) [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. ROS can damage cellular membranes, nucleic acids, proteins, and carbohydrates [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], but their toxic effects are counteracted by enzymatic and non-enzymatic antioxidative defense systems [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Salinity tolerance in plants is influenced by stress adaptation effectors [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Although quinoa exhibits higher salinity tolerance than other cereals, exposure to high NaCl concentrations can still have detrimental effects. While plants possess endogenous mechanisms to cope with salinity stress, external interventions are often necessary [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eGiven quinoa's nutritional value, various strategies, including the application of nanoparticles (NPs), have been explored to enhance its salinity tolerance, facilitating cultivation in saline soils. Research indicates that NPs can maintain crucial biochemical and physiological functions in plants, as demonstrated in cotton [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] and basil [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eZinc (Zn) and silicon (Si) play essential roles in plant physiology. Zn is involved in enzyme activation, ion homeostasis, and overall plant growth, development, and yield [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Si enhances growth, yield, crop quality, photosynthesis, nitrogen fixation, and stress tolerance [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Furthermore, graphene oxide has been found to improve chlorophyll content, relative water content, and growth performance in plants such as \u003cem\u003eSilybum marianum\u003c/em\u003e [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. This study aimed to synthesize a high-performance nanocomposite (ZnO/SiO2@GO) by decorating ZnO and SiO2 NPs onto GO and evaluate its effectiveness in mitigating the detrimental effects of high salt stress on quinoa. Seeds primed with ZnO/SiO2@GO were cultivated, and the resulting seedlings were exposed to elevated NaCl levels. The impacts on growth parameters, pigments, sugars, proteins, MDA, H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, phenolics, and peroxidase and catalase activities were assessed to determine whether the nanocomposite could enhance quinoa's tolerance to salinity, positioning it as a promising protective agent against high salt stress.\u003c/p\u003e"},{"header":"2 Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Synthesis of zinc oxide/silicon dioxide@graphene oxide nanocomposite (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO)\u003c/h2\u003e \u003cp\u003eZnO was synthesized by gradual addition of KOH solution into 0.2 M Zn(NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e solution while being vigorously stirred and centrifuged at 5000 \u0026times;g for 15 min. The resulting ZnO NPs were calcined at 500\u0026deg;C for 3 h [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].SiO\u003csub\u003e2\u003c/sub\u003ewas synthesized by dropwise addition of Na\u003csub\u003e2\u003c/sub\u003eSiO\u003csub\u003e3\u003c/sub\u003esolution to 33% ammonia/absolute ethanol (3:1 v/v) and centrifuged at 5000 \u0026times;g for 15 min. SiO\u003csub\u003e2\u003c/sub\u003e NPs were dried at 50\u0026deg;C for 24 h and 185\u0026deg;C for additional 5 h [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. GO was synthesized from the oxidation of graphite [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] using H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e/HNO\u003csub\u003e3\u003c/sub\u003e (3:1 v/v) in ice-water bath with agitation for 3 hat 35\u0026deg;C then distilled water was gradually added while being vigorously stirred. After changing the color into dark brown, the reaction mixture was diluted then H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003ewas added dropwise. The resultant GO was adjusted to pH 7and centrifuged for 15 min then GO was dried at 80\u0026deg;C. Decoration of ZnO NPs and SiO2 NPs onto GO was performed by dispersing GO in chitosan/polyvinyl alcohol (CS/PVA) [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] previously prepared from dissolving PVA in 5% acetic acid followed by the addition of CS [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. CS/PVA containing GO was ultrasonicated for an hour at 25\u0026deg;C. ZnO NPs and SiO\u003csub\u003e2\u003c/sub\u003eNPs were added and stirred for an hour, and then glutaraldehyde was added to the stirring suspension.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Plant material and growth conditions\u003c/h2\u003e \u003cp\u003eCertified seeds of quinoa [\u003cem\u003eChenopodium quinoa\u003c/em\u003e (L.)] were obtained from Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt. Seeds were surface sterilized in 1% w/v sodium hypochlorite for 5 min, rinsed with water several times and soaked for 6 h either in water or in ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO (40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). Then the seeds of both groups were planted in plastic pots (40 \u0026times; 25 \u0026times; 15 cm), each contained 3 kg of soil (clay: sand; 2:1 w/w), approximately 2 cm-depth and spaced 5 cm apart in rows and allowed to grow at 22\u0026thinsp;\u0026plusmn;\u0026thinsp;2/12\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C, day/night temperature with a 12-h photoperiod at 420\u0026ndash;460 \u0026micro;mol m\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e s\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e PPFD. All pots received water (200 ml per pot) daily for 3 days followed by Long Ashton nutrient solution up to the 10th day, then pots of both groups were divided into four subgroups for treatment with NaCl at 0 mM (control), 200 mM, 400 mM and 500 mM every 3 days (200 ml per pot) for the following 21 days. When seedlings were 35-day-old, samples were collected, washed thoroughly with water, plotted dry between layers of tissue, and separated into shoots and roots. Shoot height, root length and fresh weights were determined, and then dried at 80\u0026deg;C for 2 days for dry weight determination from which water contents were calculated. The new leaves and the upper parts were frozen in liquid N and used for analyses.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Determination of photosynthetic pigments, protein and soluble sugar\u003c/h2\u003e \u003cp\u003eChlorophylls (a and b) and carotenoids in fresh tissues were extracted with 80% acetone, then absorbance was read at 663.2, 646.8 and 470 nm. Pigment contents were calculated as \u0026micro;g ml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e then converted to \u0026micro;g unit weight\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Protein in fresh tissue was extracted with chilled acetone, centrifuged and the residues were allowed to dry at room temperature to obtain acetone powders [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Aliquots of the acetone powders were mixed with Tris-HCl (0.05 M, pH 9.0) for 10 min at 4\u0026deg;C and then centrifuged at 20000 \u0026times;g for 15 min at 4\u0026deg;C. Protein content was determined spectrophotometrically at 595 nm by reaction with Coomassie Brilliant Blue G-250 [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].Soluble sugars were extracted in 80% ethanol [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. An aliquot was mixed with 3 ml anthrone reagent (8.6 mM anthrone in 80% H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e) and heated for 10 min, then cooled in ice bath for 30 min and absorbance was read at 623 nm [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The residues remained from extraction of soluble sugars were digested in 1.6 M perchloric acid for the determination of non-soluble sugars using anthrone.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Determination of lipid peroxidation (malondialdehyde, MDA), hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) and phenolic compounds\u003c/h2\u003e \u003cp\u003eMDA in fresh tissues was extracted with 10% Trichloroacetic acid (TCA, w/v) then centrifuged at 12000 \u0026times;g for 15 min and absorbance was read at 450, 532, and 600 nm after reaction with tribromo-arsenazo colorimetry using 10% TCA containing 0.5% thiobarbituric acid from the equation: 6.45(A532\u0026thinsp;\u0026minus;\u0026thinsp;A600)\u0026thinsp;\u0026minus;\u0026thinsp;0.56(A450) [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e was extracted with 0.1% TCA and centrifuged at 12000 \u0026times;g for 15 min at 4\u0026deg;C, assayed in potassium phosphate buffer (10 mM, pH 7.0) containing 1 M KI and absorbance was measured at 390 nm [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Phenolic compounds were determined using Folin\u0026ndash;Ciocalteu reagent and absorbance was read at 765 nm [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Extraction and assay of peroxidase and catalase\u003c/h2\u003e \u003cp\u003ePeroxidase and catalase in fresh tissues were extracted with 50 mM sodium phosphate buffer (pH 7.0) containing 1 mM EDTANa\u003csub\u003e2\u003c/sub\u003e and 0.5% Polyvinylpolypyrrolidone under cold conditions and centrifuged at 13000 \u0026times;g for 30 min at 4\u0026deg;C. Peroxidase activity was assayed following the ascorbate oxidation by the decrement in absorption at 290 nm [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Catalase activity was assayed by controlling the H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003eabsorbance reduction at 240 nm [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Statistical analysis\u003c/h2\u003e \u003cp\u003eThe experiment was repeated twice and samples for each analysis were taken in triplicates from both experiments (n\u0026thinsp;=\u0026thinsp;6) and mean values (\u0026plusmn;\u0026thinsp;SD) were applied. The experiment was designed as a complete randomized block consisting of 160 pots (2 sets for ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO treatments: control, 40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) x (4 sets for NaCl treatments: control, 200 mM, 400 mM and 500 mM) x (10 replications) x (2 repetitions). The full data were subjected to analysis of variance (ANOVA) followed thereafter by least significant differences (LSD) at 5% level.\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results","content":"\u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows that NaCl concentrations of 400 mM and 500 mM significantly reduced the shoot height and root length of 35-day-old quinoa seedlings compared to control values. However, the reduction caused by 200 mM NaCl was not significant. The extent of the decrease increased with higher NaCl concentrations. Similar significant reductions were observed in fresh and dry weights as well as in the water content of both shoots and roots, with all NaCl concentrations affecting these parameters. Overall, the reductions in shoot height, root length, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, shoot water content, and root water content ranged from 18\u0026ndash;56%, 10\u0026ndash;36%, 34\u0026ndash;68%, 38\u0026ndash;79%, 16\u0026ndash;40%, 27\u0026ndash;66%, 4\u0026ndash;12%, and 3\u0026ndash;11%, respectively. However, pre-soaking quinoa seeds in the synthesized ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO nanocomposite (40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) improved the growth parameters under normal conditions, resulting in increases of approximately 17%, 11\u0026ndash;13%, and 5\u0026ndash;11% in shoot and root length, fresh weight, and dry weight, respectively, along with slight increases in water content. Furthermore, the nanocomposite completely mitigated the negative effects of all NaCl concentrations on these growth parameters, making NaCl have no significant effect except for the 500 mM concentration, which still had a significant effect on shoot fresh weight. The highest decreases induced by NaCl in the presence of the nanocomposite were limited to no more than 13%, 11%, 6%, and 1% in shoot and root length, fresh weight, dry weight, and water content, respectively, compared to the control. In contrast, without the nanocomposite, the reductions reached 56%, 79%, 66%, and 12%, respectively.\u003c/p\u003e \u003cp\u003eFigure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows that NaCl treatment significantly reduced the contents of chlorophylls and carotenoids in quinoa compared to untreated controls. The decreases induced by 200 mM NaCl were non-significant, but they increased as NaCl concentration rose. The reductions in chlorophyll a, chlorophyll b, and carotenoids at 400 mM NaCl were approximately 38%, 39%, and 35%, respectively, compared to the untreated control, while 500 mM NaCl caused reductions of 50%, 40%, and 45%, respectively. Total pigments were also significantly reduced by NaCl, with high concentrations leading to reductions of 37\u0026ndash;46%. The application of ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO under normal conditions resulted in increases of approximately 8\u0026ndash;11% in chlorophyll and carotenoid content relative to the control. In NaCl-treated samples, the nanocomposite greatly mitigated these decreases, bringing pigment levels closer to those of the untreated control. In fact, some increases were observed, particularly at 200 mM and 400 mM NaCl (2\u0026ndash;8%).\u003c/p\u003e \u003cp\u003eIn Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, protein content was significantly decreased by 400 mM and 500 mM NaCl, with the decrease being about 35% and 46%, respectively, compared to the untreated control. The decrease induced by 200 mM NaCl was insignificant. Conversely, NaCl treatment caused significant increases in soluble sugars, with increases ranging from 22\u0026ndash;24% for all concentrations. Insoluble sugar content was significantly reduced by 18%-48%, with the highest reduction observed at 500 mM NaCl. Consequently, total sugar content decreased under all NaCl treatments, with the greatest reduction (27%) occurring at 500 mM, compared to 16% and 6% reductions at 200 mM and 400 mM, respectively.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eHowever, pre-soaking quinoa seeds in ZnO/SiO₂@GO nanocomposite as a seed pretreatment greatly increased protein content by about 21% in the absence of salinity and counteracted the effects of NaCl. Additional increases in protein content of 15%, 12%, and no change were observed in response to 200 mM, 400 mM, and 500 mM NaCl, respectively. Similarly, the nanocomposite mitigated the increases in soluble sugars, returning them to levels near the control, while the decreases in insoluble and total sugars were largely compensated, with reductions no greater than 5%.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e contents were significantly elevated in NaCl-treated quinoa, with higher concentrations leading to greater accumulation. The increases induced by 200 mM, 400 mM, and 500 mM NaCl were approximately 125%, 160%, and 184% for MDA, and 103%, 147%, and 159% for H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, respectively. In contrast, NaCl treatment caused significant decreases in the endogenous contents of phenolics, with reductions of about 29%, 42%, and 49% for 200 mM, 400 mM, and 500 mM NaCl, respectively. However, pre-soaking quinoa seeds in ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO greatly suppressed the accumulation of MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, fully mitigating the effects of NaCl. Under normal conditions, the nanocomposite induced a 10% increase in phenolics, and it limited the decrease induced by the highest NaCl concentration to no more than 3%.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn concomitant, NaCl treatment significantly inhibited the activities of peroxidase and catalase, with the effect being more pronounced at 500 mM NaCl compared to 400 mM and 200 mM (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). The inhibition reached about 51\u0026thinsp;\u0026minus;\u0026thinsp;31% for peroxidase and 49\u0026thinsp;\u0026minus;\u0026thinsp;28% for catalase activities. However, seed soaking in ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO led to substantial increases in both peroxidase and catalase activities in normally grown samples (about 17% and 25%, respectively). Additionally, the inhibitory effects of NaCl on peroxidase and catalase activities were largely mitigated, and further enhancements were observed, with increases of 5%-11% for peroxidase and 6%-17% for catalase compared to control values.\u003c/p\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eQuinoa's exceptional nutritive profile and its higher tolerance to salinity compared to other cereals make it an ideal candidate for cultivation in saline and marginal soils. The ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO nanocomposite was synthesized in this study to modulate and enhance quinoa's tolerance to high salinity, providing an efficient protectant against elevated salt levels. High NaCl concentrations (400 mM and 500 mM) significantly reduced quinoa growth, photosynthetic pigments, protein, insoluble and total sugars, and phenolic compounds while inhibiting peroxidase and catalase activities. In contrast, soluble sugars, MDA, and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e levels were elevated under high salinity. These results confirm the negative effects of high salinity on quinoa growth, despite its higher salt tolerance than wheat, corn, and barley [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Generally, salinity disrupts several plant processes by altering water potential, causing ion imbalance and toxicity, impairing cell division and expansion, and generating reactive oxygen species (ROS), which further damage cellular components [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eSalinity may reduce plant growth by causing chlorophyll degradation, as chlorophyll is a critical determinant for photosynthesis. This study supports findings by Mazumder et al. [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], who observed that salt stress led to pigment degradation due to the vulnerability of photosystems. Salinity can also disrupt electron transport in the photosynthetic system, leading to oxidative damage. Carotenoids, however, protect chlorophyll from photooxidation by removing oxygen from excited chlorophyll-oxygen complexes. According to Nemat Alla et al. [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e], the inhibition of carotenoids in maize seedlings resulted in chlorophyll degradation, which compromised the photosynthetic capacity. Similarly, in this study, NaCl treatment reduced carotenoid content, potentially exacerbating the chlorophyll degradation and negatively impacting photosynthesis. However, NaCl increased soluble sugars to act as osmolytes against salinity for membrane protection and osmotic adjustment.\u003c/p\u003e \u003cp\u003eSalinity increased soluble sugars, which act as osmolytes to protect membranes and facilitate osmotic adjustment to survive extreme osmotic stress. These sugars might have resulted from the hydrolysis of polysaccharides rather than new synthesis. Elevated soluble sugars provide protection against salinity by stabilizing proteins and cell membranes under osmotic stress [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Conversely, protein synthesis appeared to be impaired by the salt-induced decrease in photosynthetic activity. This is supported by the alterations in peroxidase and catalase activities, which are vital antioxidant enzymes that protect against reactive oxygen species (ROS) generated under stress which are destructive to proteins and DNA.\u003c/p\u003e \u003cp\u003eThe substantial increase in MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e levels under NaCl treatment, coupled with inhibited antioxidant enzyme activity, suggests the induction of oxidative stress. Under oxidative stress, ROS damage proteins, DNA, and lipids, further impairing growth and metabolism. Peroxidase and catalase can scavenge H\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003eO\u003csub\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sub\u003e [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The inhibition of peroxidase and catalase activities under salt stress impedes ROS detoxification, exacerbating cellular damage and limiting growth.\u003c/p\u003e \u003cp\u003eThe present work declares that quinoa tolerated low concentrations of NaCl but suffers from the high concentration which resulted in deteriorations. However, soaking quinoa seeds in the ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO nanocomposite prior to salinity treatment improved growth and mitigated the adverse effects of NaCl. The nanocomposite enhanced antioxidant activities, alleviated oxidative stress, and improved pigment content, protein synthesis, and photosynthetic capacity. This suggests that ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO plays a key role in enhancing quinoa's tolerance to high salinity.\u003c/p\u003e \u003cp\u003eThe inclusion of Zn, SiO\u003csub\u003e2\u003c/sub\u003e, and graphene oxide (GO) in the nanocomposite likely contributed to improved plant health. Zn supports enzymatic functions and ion homeostasis [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], SiO\u003csub\u003e2\u003c/sub\u003e provides stress tolerance and boosts photosynthesis [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], and GO improves chlorophyll content and water retention [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Nanoparticles (NPs) have been reported to increase plant stress tolerance by preserving critical physiological and biochemical functions Kalteh et al. [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e] found that NPs increased growth, yield, and starch content in wheat subjected to drought stress. In consistent with the present results, NPs mitigated abiotic stresses and improved wheat growth under stress conditions [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Also, NPs increased plant tolerance and improved physiological processes of flaxseed under stress [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn this study, the nanocomposite ameliorated quinoa growth and alleviated the reduction in chlorophyll and carotenoid levels caused by NaCl, possibly by reducing oxidative stress and enhancing the synthesis of these pigments. The nanocomposite also mitigated the accumulation of MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, and increased the activity of antioxidant enzymes, highlighting its role in protecting quinoa from oxidative damage under saline conditions. These improvements in photosynthetic capacity, combined with elevated pigment content, total sugars, and protein levels, point to enhanced plant growth under stress. Zhan et al. [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] who concluded that NPs promoted wheat growth by elevating chlorophyll content. Mehrian et al. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] indicated that NPs significantly elevated redox enzyme activity in \u003cem\u003eLycopersicon esculentum\u003c/em\u003e. Similarly, Saha and Gupta [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e] reported that Ag NPs enhanced \u003cem\u003ein vitro Swertia chirata\u003c/em\u003e growth by the activation of antioxidants for scavenging free radicals as a defense mechanism in response to abiotic stress [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. So, the amelioration in quinoa growth by the nanocomposite under high salt stress synchronized with elevated photosynthetic pigments, total sugars and protein contents could reveal an improvement of photosynthetic incapacity. Meanwhile, the nanocomposite suppressed the accumulated MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e concomitantly with enhancing phenolic compounds and the activities of peroxidase and catalase pointing at ameliorated antioxidant performance that would result in modulating quinoa tolerance to high salinity and improving growth.\u003c/p\u003e"},{"header":"5 Conclusions","content":"\u003cp\u003eThe high concentrations of NaCl significantly impacted quinoa growth, pigments, protein, and sugar content, and induced oxidative stress. MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e levels increased, while phenolic compounds and antioxidant enzyme activities were inhibited. These findings clearly declare that quinoa suffered from high salinity levels. However, the application of a synthesized ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO nanocomposite (40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) through seed soaking effectively improved quinoa's growth parameters, pigment content, and protein synthesis, demonstrating its potential to alleviate the harmful effects of high salinity. The nanocomposite seemed beneficial in eliminating the oxidative stress by suppressing MDA and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e levels and enhancing phenolic compounds and antioxidant enzyme activities, making it a promising strategy for improving quinoa tolerance to high salinity. These findings highlight the nanocomposite's ability to enhance quinoa's resilience under saline conditions, which is crucial for growing quinoa in saline-affected soils.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMN: planed the scientific experiments, performed data analysis, wrote, edited and reviewed the manuscript; EB, NM and ME: performed the experiments. All the authors have approved the final manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe plant seeds used in this study were obtained from Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt. The plants used in the study comply with local guidelines. The plant samples collected in this study were cultivated varieties and planted in the greenhouse of the Faculty of Science, Damietta University, Egypt.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSamples are allowed to be picked for scientific research purposes.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSeeds were obtained from Sakha Agricultural Research Station, Kafr El-Sheikh, Egypt.\u0026nbsp;All the authors have approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there is no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOpen access funding is provided by The Science, Technology \u0026amp; Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. This work does not involve patient(s).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData availability has been declared in the \u0026lsquo;Declaration\u0026rsquo; in the system. Data availability statement that was provided in the manuscript has been added on the editorial system as well. All data supporting the findings of this study are available within the paper and its Supplementary Information.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003ePereira E, Encina-Zelada C, Barros L, Gonzales-Barron U, Cadavez V, Ferreira IC (2019) Chemical and nutritional characterization of Chenopodium quinoa Wild (quinoa) grains: A good alternative to nutritious food. 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Molecules 25:5511\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-life","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Life](https://link.springer.com/journal/11084)","snPcode":"11084","submissionUrl":"https://submission.springernature.com/new-submission/11084/3","title":"Discover Life","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"catalase, growth, peroxidase, phenolics, salinity, nanocomposite","lastPublishedDoi":"10.21203/rs.3.rs-6165086/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6165086/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe nanocomposite zinc oxide/silicon dioxide@graphene oxide (ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO) was synthesized by the decoration of ZnO nanoparticles (NPs) and SiO\u003csub\u003e2\u003c/sub\u003e NPs onto GO. The synthesized nanocomposite was used as seed soaking for modulation of tolerance of quinoa (\u003cem\u003eChenopodium quinoa\u003c/em\u003e) to high salinity stress. Seeds were soaked in 40 mg ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of the nanocomposite for 6 h thereafter NaCl was applied at 200 mM, 400 mM and 500 mM -on the 10th day of sowing- for a duration of 21 days. The results demonstrated that high NaCl (400 mM and 500 mM) significantly reduced growth parameters, photosynthetic pigments, protein, insoluble and total sugars, phenolics and activities of peroxidase and catalase. Conversely, soluble sugars, lipid peroxide (malondialdehyde, MDA) and hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) levels were elevated, with the effects being more pronounced at higher NaCl concentrations. However, using ZnO/SiO\u003csub\u003e2\u003c/sub\u003e@GO as a seed priming agent effectively mitigated the adverse effects of NaCl on these parameters. Growth was enhanced, while reductions in pigments, protein, sugars, phenolics, and enzymatic activities were counteracted. Additionally, elevated levels of soluble sugars, MDA, and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e were suppressed, approaching control levels. These findings highlight the severe impacts of high salinity stress and indicate that treatment with the ZnO/SiO2@GO nanocomposite can mitigate these adverse effects. Therefore, this nanocomposite could be considered a safe and efficient protectant to enhance quinoa's tolerance to high NaCl levels.\u003c/p\u003e","manuscriptTitle":"The synthesized zinc oxide/silicon dioxide@graphene oxidenanocomposite (ZnO/SiO2@GO) enhances quinoa tolerance to high salinity","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-28 07:54:43","doi":"10.21203/rs.3.rs-6165086/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-04-28T07:12:19+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-27T14:48:32+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-26T15:01:14+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"279758395182188755851590592191098078169","date":"2025-04-19T05:02:59+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"315844993803128318123272899662983713587","date":"2025-04-18T13:45:37+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-04-18T09:22:52+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-04-03T08:20:42+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-04-03T03:18:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-04-02T09:05:37+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Life","date":"2025-04-02T09:04:28+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-life","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Life](https://link.springer.com/journal/11084)","snPcode":"11084","submissionUrl":"https://submission.springernature.com/new-submission/11084/3","title":"Discover Life","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"987613fd-f4b3-4539-a4aa-675fd2ea50c2","owner":[],"postedDate":"April 28th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-06-30T10:08:31+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-28 07:54:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6165086","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6165086","identity":"rs-6165086","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}