Nano-priming of Phaseolus vulgaris OTI cultivar with cobalt ferrite nanoparticles enhances the mineral composition of progeny seeds

Nano-priming of Phaseolus vulgaris OTI cultivar with cobalt ferrite nanoparticles enhances the mineral composition of progeny seeds | 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 Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Nano-priming of Phaseolus vulgaris OTI cultivar with cobalt ferrite nanoparticles enhances the mineral composition of progeny seeds Yazmín Stefani Perea-Vélez, Rogelio Carrillo-González, Ma. Carmen A. González-Chávez, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4578599/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Aug, 2024 Read the published version in Journal of Nanoparticle Research → Version 1 posted 7 You are reading this latest preprint version Abstract Nano-priming is an emerging application of nanotechnology in agriculture intending to increase crop yield and nutritional quality, while reducing fertilizer applications. This study aimed to investigate the effects of seed priming with citrate-coated CoFe 2 O 4 nanoparticles (NPs) suspensions (10, 20, and 40 mg NPs L − 1 ) on the life cycle of the Phaseolus vulgaris L. OTI cultivar and evaluate the technology costs. The effect of nano-priming was assessed in the germination, flowering, and harvest stages. Unprimed and hydro-primed seeds were negative and positive controls, respectively. Nano-priming with CoFe 2 O 4 NPs had no effect neither on the germination nor on plant nutrition (in the flowering stage) of OTI beans compared to unprimed and hydro-primed seeds. In contrast, nitrogenase activity (343.3 ± 1.1 µmol h − 1 plant − 1 of C 2 H 4 ) was detected in the plants from the 40 mg kg − 1 nano-primed seeds. The K concentration of progeny seeds from nano-priming with 10, 20, and 40 mg NPs L − 1 increased significantly by 3%, 16%, and 13% compared to the control seeds. The Zn concentration in the seeds from nano-priming with 10 mg NPs L − 1 was 27% higher than in the control and 28% higher than in the hydro-primed seeds. When nano-priming with 40 mg NPs L − 1 , the Zn concentration was 5% and 6% higher than the control and hydro-primed seeds. The calculated cost of nano-priming seeds per ha ranged from 121 to 143 USD. In this regard, nano-priming of bean seeds with citrate-coated CoFe 2 O 4 NPs could be a low-cost approach to achieve nutritional security and agricultural sustainability. seed priming nano-enabled agriculture nanotechnology cost biofortification Figures Figure 1 Figure 2 Figure 3 Introduction Adopting a sustainable model that is profitable, productive, and conserves natural resources is a challenge for agriculture today, and the soils are one of the most relevant elements in this new agricultural model. The premise is that it is no longer enough to maintain its quality, but it is also necessary to be able to improve it. In this regard, low-input technologies (such as seed priming, cover crops/green manure, or intercrops) can be one of the several possible ways to achieve sustainability [ 1 ]. On the one hand, nano-enabled agriculture has shown that through nano-fertilizers it is possible to improve plant health, nutrient biofortification, increased photosynthetic output, and higher rates of nitrogen fixation[ 2 ]. Among the nano-enabled technologies, seed priming or nano-priming is an emerging field proven to be more promising than traditional priming approaches [ 3 – 5 ]. Seed priming is a low-input pre-sowing seed treatment. It allows synchronized germination and maturity of crops, improves germination time and water use efficiency, and increases nutrient uptake and tolerance of plants to biotic and abiotic stresses [ 4 , 6 ]. Seed priming is generally defined as the controlled hydration of seeds to a level that allows pre-germinative metabolic activity without radical protrusions [ 7 , 8 ]. After hydration, the seed is dried again to its original weight [ 8 , 9 ]. Unlike traditional seed priming, which employs water or solutions containing nutrients, hormones, microorganisms, or biopolymers [ 6 ], nano-priming uses nanoparticle (NPs) suspensions or nano-formulations [ 3 ]. Numerous nanomaterials have been tested as priming agents, for instance, Ag, Au, Cu, TiO 2 , FeS 2 , Fe 2 O 3 , and ZnO [ 3 , 5 , 10 – 13 ], proving that they can significantly increase the germination rate, vigor index, root elongation, plumule length, and the antioxidant response of seedling of many crops [ 5 ]. However, ferrite NPs (such as CoFe 2 O 4 , NiFe 2 O 4 , MnFe 2 O 4, and Fe 2 O 4 ) have attracted attention for nano-enabled agriculture due to their biocompatibility, and safety for humans and plants. For instance, Sundaria et al. [ 14 ] reported increased germination percentage and shoot length in two wheat varieties (high and low iron efficiency) after nano-priming with Fe 2 O 4 NPs. Cobalt ferrite (CoFe 2 O 4 ) and MnFe 2 O 4 NPs can significantly enhance the symbiotic nitrogen fixation efficiency by 260% [ 15 ], and increase the root nodulation by 61% in Glycine max (L.) Merr. compared to the control plants [ 16 ]. On the other hand, Phaseolus vulgaris (common bean) was selected for this study because is a protein source for more than 300 million people from Latin America, the Caribbean, and Eastern Africa [ 17 ]. It is considered a nearly perfect food because of its high protein, fiber, prebiotic, vitamin (A, C, and folate), and mineral content (Ca, Mg, K, Cu, Fe, and Zn) [ 18 ]. For this reason, beans are a tool for fighting to hide hunger. Another interesting aspect to consider is that the common bean is a crop that enriches the soil through biological nitrogen fixation, which is based on symbiosis with bacteria such as Rhizobium leguminosarium bv. Phaseoli [ 19 ]. The use of leguminous as intercrop in arable systems can reduce nitrous oxide emission by 18%, and N fertilizer use by 24% compared to systems without legumes [ 20 ]. Although the common bean is considered an inefficient N fixer compared to other legumes [ 21 ], bean plants are used as an intercrop in Africa by smallholder farmers. In this context, this study aimed to evaluate the effects of nano-priming on the life cycle of Phaseolus vulgaris L. OTI cultivar with citrate-coated cobalt ferrite (CoFe 2 O 4 ) NPs. We hypothesized that (1) nano-priming may improve germination traits compared to unprimed seeds, (2) enhance yield, and (3) improve the nutritional quality of bean grains. Moreover, the cost of the nano-priming was estimated. The results of this research can help to provide accessible solutions to smallholder farmers, because the low productivity of these farmers is mainly related to the limited use of inputs, such as irrigation, pesticides, mechanization, and fertilizers [ 22 ]. In this regard, nano-priming can be an alternative to reduce the dependency on external fertilizer purchases. On the other hand, the use of NPs to increase the nodulation, and N fixation capacity of beans is a breakthrough attempt at sustainable agriculture. Material and methods Two experiments were conducted to assess the effect of nano-priming with citrate-coated CoFe 2 O 4 NPs on the life cycle of P. vulgaris OTI cultivar. The first experiment evaluated the effect of nano-priming on the germination stage, while the second experiment focused on its effects from the vegetative to maturity stages of bean plants. For quality assurance and quality control in analytical measurements, the material was washed in a 0.25 M HCl solution to remove any traces of contaminating materials. All the reagents used were of analytical grade (J.T. Baker, Merck, and Sigma–Aldrich), and standard solutions were prepared using certified stock solutions. Plant material P. vulgaris OTI cultivar seeds were obtained from the Genetic Resources and Productivity Program of Colegio de Postgraduados. OTI is an improved bean cultivar that is adapted to the central high valleys of the Mexican Republic. The main characteristics of OTI beans are the cooking time (61–85 min), its yield (2.5 t ha − 1 ), and its high resistance to Colletotrichum lindemuthianum , Sclerotinia sp., and Rhizoctonia solani ; medium resistance to Uromyces appendiculathus var. appendiculatus and moderate tolerance to Pseudomonas phaceolicola . The seed coat of OTI beans has a light brown background with bright purple spots [ 23 ]. Synthesis and properties of citrate-coated CoFe 2 O 4 NPs Citrate-coated CoFe 2 O 4 NPs were synthesized using a co-precipitation method. Eight mL of 1 M Fe(NO 3 ) 3 and 4 mL of 1M Co(NO 3 ) 2 solutions were mixed. Then, 38 mL of 1.5 M NaOH solution was added dropwise to the metal ion solution under constant stirring conditions (400 rpm). The mixture solution was continuously stirred and heated at 90°C for 1h. The resulting black precipitate was washed three times with deionized water. The precipitate was then mixed with 50 mL sodium citrate 4 mM solution and stirred at 80°C for 30 min. After that, the NPs were washed with deionized water until the pH of the supernatant was 7 [ 24 , 25 ]. The washed NPs were suspended in 50 mL of deionized water to prepare a NPs stock solution. The NPs characterization data were published by Perea-Vélez et al. (2022), with results indicating that the NPs had a semi-spherical shape with an average primary size of 13 ± 5 nm, hydrodynamic diameter of 216 ± 10 nm, zeta potential in deionized water of 10.5 ± 6.6 mV, point of zero charge of 6.8, and 48% Fe, 29% Co composition. Experiment one. Assessment of the effect of nano-priming on the germination of bean The seed water uptake curve was evaluated before the start of the experiment (Figure S1 ). This determined the imbibition time for nano-priming because each crop cultivar has its critical soaking duration, and the success of priming depends on achieving optimal seed hydration. So, according to this assay, the soaking time was fixed at 2 h and 30 min. Afterward, a completely randomized experimental design was set up to assess the effect of nano-priming on the germination traits of the P. vulgaris OTI cultivar. The treatment factor was the concentration of the NPs suspension with three levels: 10, 20, and 40 mg NPs L − 1 (equivalent to 3.5, 7.5, and 15 mg Fe L − 1 , and 2.4, 4.5. and 9.5 mg Co L − 1 , respectively). The concentrations of the nano-priming suspensions were chosen to ensure the dispersion of NPs and to avoid their fast agglomeration. Furthermore, to differentiate between water and NPs effects, seeds were primed with distilled water (hydro-priming or positive control), and unprimed seeds were used as a negative control. Three replicates were used for each treatment (clear-hinged containers with ten seeds). Preparation of priming suspension and nano-priming The priming suspensions were freshly prepared in sterilized deionized water before their use. An aliquot of NPs stock solution was placed in a Flask to get a NPs concentration of 10, 20, and 40 mg NPs L − 1 . The NPs suspension was sonicated with an ultrasonic probe for 2 min at 130 W and 90% amplitude. On the other hand, bean seeds were surface-sterilized by soaking them in a 3% (v/v) sodium hypochlorite solution for 10 min and then carefully rinsed with sterilized deionized water to remove all chlorine. Afterward, the seeds were soaked in the NPs suspension for 2 h 30 min at room temperature (20–25°C) with constant agitation on a rotor shaker (30 turns per minute). For the hydro-priming, distilled water was used instead of NPs suspension. The primed seeds were then dried back to their original moisture content; for this, the seeds were placed in sterilized paper bags and dried in an oven at 25°C for 3 d. Dried seeds were then stored at 4°C for fifteen days until further use. Seed germination assay Ten seeds were placed in a clear-hinged plastic container that was surface-disinfected. The bottom of the container was lined with sterilized cotton and two sterile filter papers. The seeds were moistened with 15 mL of sterile distilled water and kept in the darkness at 24°C for 7 d to initiate germination. Germinating seeds were observed and counted daily based on radical emergence up to 2 mm [ 12 ]. Germination percentage (GP), energy period (EP), germination energy (GE), germination rate (GR), and mean germination time (MGT) were calculated based on the following equations [ 27 , 28 ]: $$GP=\frac{number of seeds germinated}{total number of seeds placed for germination test}\times 100$$ $$EP=Days after starting the germination test to reach \ge 50\text{%} of germinated seeds$$ $$GE=\frac{Cumulative daily total of germinated seeds at day 4}{Number of seeds set to germinate}\times 100$$ $$GR=\left(\frac{a}{1}\right)+\left(\frac{b-a}{2}\right)+\left(\frac{c-b}{3}\right)+\dots +\left(\frac{n-{n}_{-1}}{D}\right)$$ $$mean germination time \left(MGT\right)=\frac{\sum \left(D\times n\right)}{\sum n}$$ where a, b, c, …, n is the number of seeds germinated on days 1, 2, 3, …, D, and D is the number of days counted from the beginning of the test. The fresh and dry weights of the seedlings were determined at the end of the germination experiment (day seven). Stem height and root length were measured using the ImageJ software. The germination index (GI) and the relative seed germination (RSG) were calculated according to the following equations [ 29 ]: $$GI=(G\times L)/(Gw\times Lw)$$ $$RSG=\frac{Seeds germinated from nanopriming }{Seeds germinated from hydro-priming or control }\times 100$$ Where G and L are the germination and radicle length recorded for a specific treatment, respectively, and Gw and Lw are the values recorded for the control seeds. The seedling vigor was calculated based on the following equation [ 27 ]: $$Vigor index =germination \text{%} \times seedling weight (root+shoot)$$ Quantitative estimation of Fe and Co content in nano-primed seeds Ground dried primed seeds (0.5 g) were acid-digested (1 mL H 2 O 2 and 4 mL HClO 4 :H 2 SO 4 ; 4:1 v/v), and the digested sample was diluted to 25 mL with deionized water. The Fe and Co concentrations were determined using flame atomic absorption spectroscopy (Perkin Elmer, model 3110). Localization of NPs and iron in nano-primed seeds The Perls’ Prussian blue staining technique was employed for the initial detection of NPs within seeds [ 30 ]. This process involved placing dried primed seeds in distilled water for one hour to hydrate the primed seeds partially. Subsequently, the primed seeds were incubated with the Prussian blue solution for one hour to allow staining. After staining, the seeds were thoroughly rinsed with distilled water, and excess water was removed using paper tissue. The samples were then mounted on microscope slides and subjected to qualitative analysis using a reflectance light microscope (Carl Zeiss Stereo V20). High-resolution images were captured by using a Canon 5D digital camera. Environmental scanning electron microscopy ESEM analysis was performed to confirm the presence of the NPs in the seeds. The dried nano-primed seeds were mounted on double-sided carbon conductive tape and observed using an ESEM (Carl Zeiss EVO LS10, Jena, Germany) microscope and an X-ray detector (Bruker, Quantax 200, Germany). Images were taken at 30 kV and 80 Pa of water vapor pressure. Experiment two. Effect of nano-priming on the growth of bean plants (from VE to R2 growth stage) and nodulation (greenhouse experiment) A completely randomized experimental design was used in this study. The treatments were as follows: negative control (unprimed seeds), positive control (hydro-primed seeds), and nano-primed seeds at 10, 20, and 40 mg NPs L − 1 . Each treatment had ten replicates. Clay soil collected from the surface layer of the experimental field of Colegio de Postgraduados (19°27’56.0” N, 98°54’ 10.4” W) was used in this experiment. The soil properties are listed in Table S1 . The soil was air-dried for at least one week in the shadow. Then, it was sieved to < 2 mm and manually homogenized with a shovel. No external source of macro or micronutrients was added to the soil because soil nutrient content was high enough to fulfill the nutrient demands of beans (53 kg N t − 1 , 8 kg P t − 1 , 55 kg K t − 1 , 40 kg Ca t − 1 , 8 kg Mg t − 1 , 271 g Fe t − 1 , 90 g Cu t − 1 , and 192 g Zn t − 1 ) according to Ayala Garay et al. [ 31 ]. Two primed or unprimed seeds were sown in pots containing 8 kg of soil. Seedling emergence was recorded when the cotyledons were completely raised above the soil. At the V2 growth stage (when the second trifoliate leaf was established), one of the seedlings was removed. The plant height was recorded once a week after plant emergence. The plants were grown in an open greenhouse from April to June 2022 at an average temperature of 21°C (maximum and minimum temperatures of 27°C and 14°C, respectively). Plants were watered daily with tap water to maintain approximately 60% of the soil field moisture capacity. To assess the effect of nano-priming on nodulation, four plants per treatment were carefully harvested 51 d after sowing (R2 plant growth stage), and shoots were separated from the roots. Shoots were used to determine the leaf area and plant nutrition, while fresh nodules were collected, and the nodule fresh weight and number of active nodules (pink pigmented) or inactive nodules (brown dark-green) per plant root were determined. Nitrogenase activity Nitrogenase activity was determined by acetylene reduction. For plants at the R2 growth stage (51 d after sowing), samples of roots with nodules were transferred into a plastic hermetic closed container (1.5 L), and the excess adhered soil was carefully removed before placing the roots in the container. A serum stopper was inserted, and 10% of the air was replaced with acetylene and incubated for 1 h [ 32 , 33 ]. The gas samples were analyzed using a gas chromatograph (Clarus 400, PerkinElmer). Foliar area Digital images (Moto g6 camera) of leaves arranged on a white background and under bright light conditions were taken. Images were analyzed using the ImageJ analysis software. Evaluation of plant nutrition The aerial part of the plant was dried in an oven at 70°C for 3 d and then ground into a powder. An aliquot of 500 mg was acid-digested (one mL H 2 O 2 and four mL HClO 4 : H 2 SO 4 , 4:1 v/v) in a digest block for 16 h at 90°C. The digested samples were then made up to 25 mL with deionized water and filtered. The Ca, Mg, Fe, and Zn concentrations were determined using flame atomic absorption spectroscopy (Perkin Elmer, model 3110). The N concentration was calculated by the Kjeldahl method [ 34 ], while the P and K concentration was determined using the vanadomolybdo phosphoric acid colorimetric method [ 35 ] and flame photometry (Jenway, PFP7), respectively. Assessment of the effects of nano-priming on the agronomic traits and grain nutritional quality of bean plants Plants (6) were harvested when they reached maturity (R9, 81 d after sowing). The agronomic variables evaluated were the number of pods per plant, weight of pods, number of seeds per pod and plant, seed yield (weight of seeds per plant), seed index (100 seed weight), and harvest index. The macro (Ca, Mg, K, and P) and micronutrient concentrations (Fe, Zn, and Cu) in the seeds were determined by atomic absorption spectroscopy (Perkin Elmer, model 3110) or flame photometry (Jenway PFP7). The P concentration in the seeds was determined using the vanamolybdo phosphoric acid colorimetric method [ 35 ]. The protein concentration in the seeds was evaluated using the Kjeldahl method [ 34 ]. The protein content was calculated by multiplying the amount of total nitrogen by a conversion factor of 6.25, which assumes that the nitrogen content of proteins in foodstuffs is 16% [ 36 ]. The phytic acid concentration in seeds was estimated using the colorimetric Wade reagent method, modified as described by Gao et al. [ 37 ]. The molar ratios of phytic acid to Fe and Zn were calculated by converting the concentrations of Fe (55.84 g mol − 1 ), Zn (65.38 g mol − 1 ), and phytic acid (660.04 g mol − 1 ) into moles. Economic evaluation of nano-priming seed treatment The economic analysis was performed using the economic indicators described by Dhaliwal et al. [ 38 ] and Sarwar et al. [ 39 ]: $$Gross income=yield \left(t {ha}^{-1}\right)\times grain price$$ $$Profitable return \left(PR\right)=gross income-total production cost$$ $$PR over control=PR-control treatment$$ $$Cost benefit ratio \left(CBR\right)=\frac{PR over control}{Total production cost}$$ $$Investment factor \left(IF\right)=\frac{Gross income}{Total production cost}$$ The estimation of the gross income indicator relied on the established guaranteed price of beans in Mexico in 2022 (MXN 16, 000 t − 1 ) as reported by Mexico’s Secretariat of Agriculture and Rural Development [ 40 ]. To calculate production costs, data from the five leading bean-producing states in Mexico were obtained from the Trust Funds for Rural Development (FIRA in Spanish; Table S2 ). The cost of the nano-priming treatment was determined by considering the price of nanoparticles, which amounted to 5 750 USD per kg [ 41 ]. To report the economic indicators in terms of USD, the exchange rate used was 19.9 MXN/USD [ 42 ], which was an average of the exchange rates between 1 and 15 June 2022. Data analysis The data were analyzed with the statistical software R version 4.0.3 [ 43 ]. Germination data were analyzed using a time-to-event model [ 44 , 45 ] because this type of analysis provides more reasonable inferences considering that the germination event did not occur at the specific time of evaluation but during the interval between assessments. Parametric and non-parametric approaches were applied for data analysis. The goodness-of-fit model was evaluated graphically from the observed versus the predicted values. In the case of the parametric time-to-event model, a likelihood ratio test was performed to compare the time-to-event models between the different treatments. Model optimization was done using the Akaike Information Criterion (AIC), where the selected model had the lowest AIC. In the case of no parametric approach to modeling the effect of the experimental factor, a Wilcoxon-type statistic was carried out [ 46 ]. Plant growth (plant height) data were analyzed using repeated-measures ANOVA. Subsequently, pairwise comparisons between time points at each and between group levels were performed using the Bonferroni p-adjustment method. On the other hand, an analysis of variance was used (α = 0.05) to compare the effect of seed priming on germination traits, agronomic traits, and seed nutritional quality. Before ANOVA analysis, Shapiro’s and Bartlett’s tests were used to verify the compliance with the assumptions of normality and homogeneity of variances. Data that did not exhibit assumptions of normality and homogeneity of variance (GP, GR, EP, GE, MGT, relative seed germination vs. control treatment, number of nodules, and fresh weight of nodules) were analyzed using the Kruskal-Wallis rank sum test. The data for phytic acid and P grain concentrations were box cox-transformed. The Tukey honest significant difference test (α = 0.05) and Bonferroni test were used to detect differences between treatments. Results and discussion Nano-priming of OTI bean and its effects on the germination variables The seed Fe concentration, after nano-priming with 10, 20, and 40 mg NPs L − 1 , were 53.31 ± 3.58, 53.72 ± 6.31, and 57.27 ± 2.99 mg kg − 1 , respectively. The seed Fe concentration was not significantly (p = 0.0571) different from that of unprimed (60.90 ± 0.22) and hydro-primed (53.73 ± 2.99) seeds. Cobalt was not detected in the acid digestion extracts of the primed seeds. After nano-priming, the NPs may or may not be taken up by the seeds, but It has been suggested that most NPs may remain on the seed surface after nano-priming [ 47 , 48 ]. Histochemical staining and microscopic analyses were used to confirm the presence and location of NPs on or within the seed. These analyses complemented the atomic absorption analysis, which did not reveal significant differences in Fe seed concentration between primed and unprimed seeds. Based on the staining intensity, the NPs were localized in the hilum (Fig. 1 b white arrows) and seed coat (Fig. 1 a white arrows). In comparison, the blue staining intensity of cotyledons increased with increasing NPs concentration (Fig. 1 c). Elemental mapping analysis also confirmed the presence of Fe and Co in the cotyledons (Fig. S2 ). These findings suggest that NPs adhere to the seed coat and are taken up by the seed through the hilum region. This is consistent with previous studies that showed that Ag, Cu, Zn, and Fe NPs remained within the common bean seed coat, mainly in the hilum area.[ 49 , 50 ] The adhesion and uptake of NPs may be attributed to their physical and chemical properties, such as size, charge, and surface chemistry [ 3 ]. The positive surface charge (zeta potential 10.5 ± 6.6 mV [ 26 ] of the citrate-coated CoFe 2 O 4 NPs favors the electrostatics interaction of NPs with the negatively charged groups on the seed coat surface [ 51 ]. However, the precise mechanism involved in the interaction between NPs and the seed coat requires further investigation to optimize the use of NPs for seed priming applications, as this aspect has received relatively little attention so far [ 11 , 49 ]. The seed water uptake after nano-priming ranged from 40%±3–48%±4%, but no significant changes (p = 0.101) were observed compared to the hydro-priming (52.3%± 5.2%). These results contrast with other studies where nano-priming promoted faster water uptake compared to conventional seed priming techniques. For example, Afzal et al. [ 52 ] reported 50% more water uptake in rice grains imbibed with 20 and 40 mg FeO NPs L − 1 for 24 h compared with hydro-priming and FeSO 4 solution (20 mg L − 1 ). Similar results were observed with other types of NPs as priming agents and other seeds. For instance, nano-priming of rapeseeds ( Brassica napus variety Zhongshuang 11) with polyacrylic acid-coated nanoceria (0.1 mM) increased the water uptake by 52% compared to the priming with TES buffer in the first hour of the priming. The increment of water uptake in nano-primed seeds after 3 and 8 h of imbibition was 14% and 12% more compared to the observed with TES buffer treatment [ 10 ]. In wheat grains, nano-priming with ZnO NPs (10 mg L − 1 ) increased water uptake by 51% compared to hydro-priming after 12 h of imbibition [ 12 ]. It has been pointed out that nano-priming accelerates the seed water uptake. However, the mechanisms of water uptake into the seed under the influence of NM are still unknown [ 53 ]. Moreover, the effects of NPs on plant or seed performance depend on the properties and concentrations of NPs, plant species, and seed properties [ 47 , 53 ]. Using traditional evaluation methods, which consider variables such as GP, GI, RSG, EP, MGT, and GR, it was observed that nano-priming did not exert a significant influence on the germination of OTI beans compared to hydro-priming and control treatments (Table S3 ). However, the time-to-event analysis revealed differences in the germination curves between treatments. Non-parametric time-to-event curves (Fig. 2 c) showed that the germination of control seeds and those nano-primed with 10 mg NPs L − 1 differed from the other treatments, as confirmed by Wilcoxon scores. The sum of the Wilcoxon scores for control and nano-primed seeds at 10 mg NPs L − 1 were 7.57 and 3.57, respectively, while negative values were observed for hydro-primed seeds (-2.08), and nano-primed seeds at 20 mg NPs L − 1 (-2.54) and 40 mg NPs L − 1 (-6.52). Thus, the germination of control and nano-primed seeds with 10 mg NPs L − 1 was, on average, the fastest compared to the other treatments. This conclusion was further supported by the parametric time-to-event curves, which estimated the lowest median germination time for control and nano-primed seeds at 10 mg of NPs L − 1 (Table 1 ). Overall, the parametric approach of time-to-event models is well-suited for making inferences about underlying biological mechanisms, such as germination [ 46 ]. However, the likelihood ratio test for the parametric log-logic time-to-event model curves (p = 0.0164) indicated that the germination curves were different between treatments (Fig. 2 b). Based on the AIC criterion, the control treatment had the fastest germination, followed by 10 mg NPs, hydro-priming, 40 mg NPs, and 20 mg NPs. The nano-priming did not affect the fresh (p = 0.066) and dry weight (p = 0.176) of seedlings, average root length (p = 0.068), root diameter (p = 0.195), or the number of roots (p = 0.313; Table S4 ). No significant differences (p = 0.061) were found between the vigor index of seedlings from the nano-priming and control (Table S4 ). These observations contrast with studies that stated that nano-priming accelerates or enhances germination and improves seedling growth and vigor in different plant species [ 53 , 54 ]. In rice, nano-priming with FeO NPs at 20 and 40 mg L − 1 increased radicle length by 50% and plumule length by 22% compared with hydro-priming [ 52 ]. Pawar et al. [ 55 ] found that Fe 2 O 3 NPs at 4–8 mg L − 1 significantly enhanced the radicle length and plumule length of chickpea ( Cicer arietinum L variety Digvijay) seedlings. Table 1 Parameter estimates from the parametric time-to-event model and the Akaike information criterion (AIC) for the germination of OTI beans after priming treatment. Parameter Treatment Control Hydro-priming 10 mg NPs 20 mg NPs 40 mg NPs Slope 27.40 (29.26) 11.77* (2.19) 14.67* (2.58) 8.21* (1.52) 12.37* (2.35) GF † 0.97* (0.03) 0.93* (0.05) 0.97* (0.03) 0.94* (0.05) 0.90* (0.05) MGT † 3.01* (0.13) 3.39* (0.11) 3.17* (0.09) 3.44* (0.15) 3.59* (0.12) AIC 90.69 106.62 94.79 123.15 108.45 † GF, germinate fraction; † MGT, mean germination time for the germinate fraction; * indicates that the estimate is significant; data in parentheses are standard errors of estimates. Effect of nano-priming on the life cycle of OTI bean plants and the agronomic traits From primed seeds, seedling emergence between 80% and 100% was observed 8 d after sowing (Fig. 3 a). One hundred % of seedling emergence was recorded 11 d after sowing. Overall, the plants (from unprimed and primed seeds) had a shorter growing duration (81–83 d after sowing) than the varietal description (110–130 d after sowing), and flowering occurred 11 d before the expected time (Fig. 3 b). The plant height (until R2, flowering) resulting from primed seeds exceeded the maximum height reported in the varietal description (Fig. 3 b). The simple main effect of nano-priming was significant at 14 d (p = 0.018) and 20 d (p = 0.013) for treatments with NPs at 10 and 20 mg L − 1 (Fig. 3 b). When comparing the plant growth curves (Fig. 3 b), plants from treated seeds with NPs at 20 and 40 mg L − 1 were taller (75.1 ± 13.0, and 68.9 ± 12.7 cm, respectively) than those of the control (43.9 ± 5.1 cm) and nano-priming with 10 mg NPs L − 1 (61.8 ± 20.0 cm). On the other hand, in terms of the vegetative growth parameters (foliar area, fresh and dry weight of shoot and root system; Table S5 ), plant performance until the flowering stage was not affected by the nano-priming compared to the control or hydro-priming treatment (p > 0.1). The nutritional status of plants at the flowering stage (R2, 51 d after sowing) was within the normal values of reference for macro-and micronutrients for soybean and other beans (Table 2 ), except for N, Ca, Mg, and Fe, the concentrations of these elements was higher than the typical values found for beans. When comparing the plants from the nano-priming with those from the control and hydro-priming, no significant differences were observed in the concentrations of macronutrients (N, P, K, Ca, and Mg), Fe, and Cu. Nevertheless, the Zn concentration in the leaves of plants treated with 20 mg NPs L − 1 was higher than in plants treated with 10 mg L − 1 of NPs. Table 2 Nutritional status of OTI bean plants at the flowering stage (R2, 51 d after sowing) and reference values. Treatment N P K Ca Mg Zn Fe Cu (%) mg kg − 1 Control 28.6 ± 5.4 ns 0.3 ± 0.0 ns 3.7 ± 0.1 ns 6.8 ± 1.6 ns 1.1 ± 0.1 ns 47.1 ± 0.3 ab 487.3 ± 152 ns 15.3 ± 2.5 ns Hydro-priming 28.8 ± 3.8 ns 0.3 ± 0.0 ns 3.8 ± 0.4 ns 6.0 ± 1.4 ns 1.0 ± 0.1 ns 47.5 ± 2.4 ab 418.5 ± 95 ns 11.4 ± 7.0 ns 10 mg NPs 28.5 ± 3.7 ns 0.3 ± 0.0 ns 3.8 ± 0.7 ns 6.6 ± 0.9 ns 0.9 ± 0.1 ns 45.1 ± 3.6 b 472.3 ± 125 ns 10.3 ± 3.4 ns 20 mg NPs 25.2 ± 2.5 ns 0.3 ± 0.0 ns 3.4 ± 0.3 ns 5.9 ± 1.5 ns 0.9 ± 0.1 ns 57.9 ± 9.4 a 305.7 ± 230 ns 12.4 ± 5.2 ns 40 mg NPs 29.3 ± 3.8 ns 0.3 ± 0.0 ns 3.3 ± 0.4 ns 4.9 ± 1.7 ns 0.9 ± 0.1 ns 47.6 ± 4.0 ab 510.6 ± 160 ns 8.1 ± 1.7 ns Reference values Sufficient or normal 4.25–6.0 † 0.30–6.00 † > 2.0 † 0.8–3.0 † > 0.30 † 25.0–150.0 δ 25.0–300.0 † 5.0–30.0 δ Mean ± standard deviation of n = 4. Different letters represent significant differences between the treatments (Tukey’s α = 0.05). and ns means that no significant differences were detected between the treatments. Reference values for interpreting plant analyses were obtained according to δ Kabata-Pendias [ 56 ] and † Plank and Kissel [ 57 ]. Regarding nodule formation at the flowering stage (51d after sowing), no significant differences were found in the number of active nodules and their fresh weights due to the treatment factor. It is worth mentioning that the observed nodules were products of native rhizobia present in the soil. The soil used has a history of growing maize plants, and symbiotic bacteria, such as Rhizobium etli , are naturally associated with the rhizosphere of maize and inside their roots [ 58 ]. In contrast, the reduction of acetylene (to ethylene C 2 H 4 ) was only detected in the roots of plants from the 40 mg kg − 1 nano-primed seeds. The produced C 2 H 4 was 343.3 ± 1.1 µmol h − 1 plant − 1 , this result is in agreement with De Souza-Torres et al. [ 21 ] who found that the application of iron oxide (Fe 3 O 4 ) NPs, through irrigation water, stimulated the nitrogenase activity (from 4.3 to 6.5 µmol h − 1 plant − 1 ) and nodulation of common bean plants compared to control plants (without NPs supplementation). Zhang et al. [ 59 ] reported that seed soaking plus leaf spraying of 10 mg L − 1 of FeNPs significantly increased the nitrogenase activity by 91% compared with the control plants of alfalfa (45 d old after sowing). On the other hand, at the R9 stage (maturation), it was observed that plants derived from nano-primed seeds with 40 mg L − 1 exhibited the highest number of total nodules (187 ± 35) compared to both the control plants (40 ± 25) and those subjected to hydro-priming (84 ± 55, Table S6 ). The maximum change in the total nodules between flowering and plant maturity stages was observed in plants from the nano-priming seeds with 20 mg L − 1 (3.38-fold change), followed by plants from the nano-priming with 40 mg L − 1 (2.74-fold change), hydro-priming (1.47-fold change), nano-priming with 10 mg L − 1 (0.36-fold change) and control treatment (0.25-fold change). However, future analysis should show that the nodules after flowering are active during this period because the root nodules have a short life span (12–18 d) [ 60 ], and there was a lapse of 28 days between flowering and harvesting. There is limited information regarding the effect of nano-priming with CoFe 2 O 4 NPs on biological nitrogen fixation, but our observations agree with other studies that yielded promising results regarding the effects of Fe 2 O 4 and Fe 3 O 4 NPs on root nodulation. For instance, Ma et. al. [ 15 ] found that the supplement of CoFe 2 O 4 at 10 mg kg − 1 in the growth medium (river sand) enlarged and increased total nodules by ~ 30%, in soybean plants ( Glycine max L. Meer “Williams 82”) compared to the control treatment (without amendment of NPs). Ma et al. [ 16 ] demonstrated that MnFe 2 O 4 NPs enhanced nodulation in soybeans, leading to a 61% increase in nodules and a 51% increase in nodule weight compared to the control (without NPs), along with improved nitrogen fixation (~ 2 to 2.5 times higher than the control). Wang et al. [ 61 ] showed a 35% increase in root nodules in soybean through foliar application of Fe 3 O 4 NPs compared with the unexposed plants. Comparisons between studies are challenging due to varying experimental conditions, crops, and NPs application methods, these findings underscore the potential positive impact of NPs on biological nitrogen fixation and nodulation. Increasing the capacity of biological nitrogen fixation can reduce or even eliminate the need for synthetic fertilizer [ 62 ] and is an effective strategy to enhance food security [ 63 ]. In this regard, nanotechnology and nano-priming may offer a pathway to improve biological nitrogen fixation, but a more comprehensive understanding of plant-bacterial-NPs interaction is essential. For the agronomic traits (Table S6 ), nano-priming and hydro-priming did not have significant effects on plant dry weight (p = 0.947), stem diameter (p = 0.754), number of pods (p = 0.448), seed yield per plant (p = 0.997), seed index (p = 0.0.692), and harvest index (p = 0.536). However, pod size, number of seeds per pod, and seed index (weight of 100 seeds) were within the values reported in the varietal description. Effect of nano-priming on the nutritional quality of OTI bean seeds Their high protein and mineral contents characterize beans compared to cereals.[ 64 ] In the present study, the protein content in all treatments ranged from 106–137%. Despite the lack of significant differences between offspring seeds from the primed seeds (nano-and hydro-primed), the control, protein concentration was higher than the reported protein content in Mexican bean varieties (14% and 33%) [ 64 ]. Nano-priming affected the mineral composition of bean seeds, as shown in Table 3 . Progeny seeds from plants grown from nano-primed seeds with 20 mg NPs L − 1 , exhibited higher K concentrations than those from the control treatment. Unexpectedly, the highest Zn seed concentration was observed in the offspring seeds from plants of primed seeds with 10 and 40 mg NPs L − 1 compared to the control, hydro-primed. The Zn concentration in the seeds was higher than the value range reported for Mexican bean varieties (Table 3 ), as well as the average Zn concentration in beans (28–31 mg kg − 1 ) from other regions around the globe [ 65 , 66 ]. The Zn concentration in beans from the prime treatments was 29% higher than the extreme values observed in beans, such as 77 mg Zn kg − 1 [ 65 , 66 ]. The target value for Zn biofortification is 17 mg kg − 1 above the local materials in each country or region [ 67 ]. Thus, offspring from the primed seeds can be considered biofortified, as the Zn concentration was between 32 and 59 mg kg − 1 above the average Zn concentration for Mexican bean varieties. Additionally, the Zn concentration in progeny seeds from the 10 and 40 mg NPs L − 1 primed seeds compared favorably with those found in animal products, where the Zn concentration ranged between 23 and 170 mg kg − 1 on a dry weight basis [ 65 ]. In contrast, no significant effects of the nano-priming treatment on the Ca, Mg, Fe, and Cu concentrations were observed in the offspring seeds. Nevertheless, the Fe seed concentrations among the treatments were higher than the average Fe concentration reported for seeds of Mexican bean varieties (35–58 mg kg − 1 ) [ 64 , 68 ]. Meanwhile, it was below the target concentration for biofortification (140 mg kg − 1 ). According to the CIAT, the target Fe concentration for beans must be at least 94 mg Fe kg − 1 above the concentration of local varieties in each country or region [ 65 ]. The potential bioavailability of Fe and Zn depends on their concentration of anti-nutritional compounds, like phytic acid (PA), polyphenols, lectins, and tannins. Among those compounds, it is suggested that PA is one of the primary and significant inhibitors of mineral bioavailability (and thus uptake) from beans, followed by polyphenols [ 69 ]. The PA concentrations ranged from 0.35 to 0.40 g 100 g − 1 for the progeny seeds of plants grown from nano-primed seeds. The PA concentration observed in the control, and primed treatments was statistically similar. However, the PA concentration of tested seeds was below the average phytic acid concentration reported for Mexican bean varieties and bean seeds around the world (from 0.4 to 2.6 g 100 g − 1 ) [ 17 ]. The relative bioavailability of Fe and Zn was determined based on the phytic acid to mineral molar ratios. In the case of Fe, the PA: Fe molar ratio was 5:1 on average, and no significant effect was found in the nano-priming treatment. Likewise, these PA: Fe ratios were lower than the range (from 6:1 to 33:1) observed in different bean seeds worldwide [ 17 ]. For Zn, the lowest ratio (3:1) was observed in seeds from the plants from primed 10 mg NPs L − 1 seeds. Both values can be interpreted as low bioavailability compared to the recommended values (1:1) for adequate bioavailability [ 70 ]. Table 3 Mineral composition, phytic acid concentration, and phytic acid: iron or zinc (phy:Fe, phy:Zn) molar ratio of OTI bean seeds from plants of primed or unprimed seeds. Variable Treatment Reference values Control Hydro-priming 10 mg NPs 20 mg NPs 40 mg NPs Mexican varieties* , δ HarvestPlust † Macronutrients (%) Ca 0.06 ± 0.01 ab 0.10 ± 0.03 ab 0.09 ± 0.02 ab 0.11 ± 0.03 a 0.10 ± 0.00 ab 0.11–0.57 Mg 0.18 ± 0.01 ns 0.17 ± 0.01 ns 0.17 ± 0.01 ns 0.18 ± 0.01 ns 0.18 ± 0.00 ns 0.11–0.13 K 1.71 ± 0.15 c 1.91 ± 0.14 abc 1.76 ± 0.06 ab 1.98 ± 0.06 a 1.93 ± 0.06 ab 0.85–0.95 P 0.05 ± 0.00 ab 0.05 ± 0.01 b 0.06 ± 0.01 ab 0.07 ± 0.01 a 0.05 ± 0.00 b 0.27–0.36 Protein 106.89 ± 14.2 ns 135.26 ± 39.1 ns 127.39 ± 12.3 ns 136.68 ± 18.8 ns 133.20 ± 18.1 ns 14–33 190–230 Micronutrients (mg kg − 1 ) Fe 67.23 ± 1.90 ns 70.71 ± 4.11 ns 62.91 ± 2.07 ns 68.13 ± 7.07 ns 74.91 ± 9.17ns 35–58 64–119 Zn 78.9 ± 14.3 b 77.8 ± 5.2b 100.1 ± 1.3 a 84.1 ± 6.8 b 82.8 ± 7.6 ab 27–41 Cu 5.4 ± 0.8 ns 4.9 ± 0.8 ns 4.2 ± 0.9 ns 4.4 ± 0.1 ns 5.0 ± 0.5 ns 1.14–2.02 Others Phytic acid (g 100g − 1 ) 0.47 ± 0.04 a 0.43 ± 0.05 ab 0.35 ± 0.08 ab 0.38 ± 0.01 ab 0.40 ± 0.07 ab 0.98–2.6 Phy:Fe ratio 5.22 ± 0.5 ns 4.87 ± 1.11 ns 4.96 ± 1.54 ns 5.10 ± 1.06 ns 4.59 ± 0.81 ns Phy:Zn ratio 5.96 ± 0.97 a 5.56 ± 0.82 a 3.51 ± 0.86 b 4.72 ± 0.93 ab 4.85 ± 0.93 ab Mean ± standard deviation of n=. Different letters represent significant differences between the treatments (Tukey’s α = 0.05), and ns means that no significant differences were detected between the treatments. Phy:Fe ratio, phytic and iron molar ratio, Phy:Zn ratio, phytic and zinc molar ratio. Reference values of the mineral composition of bean seeds for Mexican varieties according to * Chávez-Mendoza and Sánchez [ 64 ], and δ Espinoza-García et al. [ 68 ]. † Reference values for protein and Fe reported for the bean varieties of the HarvestPlus program [ 71 ]. Cost, benefits, and challenges of nano-priming seed treatment The gross income estimated for the seed priming (including hydro- and nano-priming) was from 3,139.75 to 3,590.79 USD per ha (Table 4 ). Meanwhile, the gross income from unprimed seeds was 3,521.79 USD. No significant differences (p = 0.83) were observed in the gross income between the control and the nano-priming. The profitable return of nano-priming over the control (unprimed seeds) was between 163.08 and 524.70 USD less. The negative profitable return of nano-priming over the unprimed control can be explained by the relatively low (but not statically significant) seed yield (Table S6 ) of plants from the nano-primed seeds. However, seed priming may be a commercially viable form of incorporating nanotechnology into agriculture compared with the soil application or foliar spray of nano-fertilizer of micronutrients because seed priming is an inexpensive and straightforward method to enhance crop quality in resource-limited areas. For example, the calculated cost of nano-priming bean seeds (with citrate-coated CoFe 2 O 4 NPs) ranged from 121 to 143 USD per ha and used a concentration suspension from 10 to 40 mg NPs L − 1 . For wheat fortification, the estimated cost ranged from 44,283 to 65,523 USD per ha for soil fertilization (98–145 mg CoFe 2 O 4 NPs kg − 1 ), and 1,553 USD for foliar fertilization per ha [ 41 ]. Table 4 Economic indicators for the economic evaluation of the nano-priming seed treatment of OTI bean per hectare of cultivated land. Treatment Gross income (USD) Cost of production (USD) Profitable return (USD) Profitable return over control (USD) Benefit-cost ratio Investment factor Control 3,521.78 936.89 2,584.89 3.76 Hydro-priming 3,590.79 937.65 2,653.14 68.25 0.07 3.83 10 mg NPs 3,479.80 1,057.99 2,421.82 -163.08 -0.15 3.29 20 mg NPs 3,421.55 1,065.18 2,356.38 -228.52 -0.21 3.21 40 mg NPs 3,139.75 1,079.56 2,060.19 -524.70 -0.49 2.91 The potential cost of nanotechnology is a crucial concern for farmers and one of the main factors considered for investing in this new technology [ 72 ]. Farmers from the wine industry of New Zealand expressed acceptance of this novel technology if the nanotechnology can significantly reduce the cost of production, shorten production time, or enhance quality without increasing cost. Indeed, they also expressed that they are willing to prioritize quality over profitability [ 72 ]. As mentioned above, nano-priming is a cheaper approach for implementing NPs in agriculture compared to the foliar or soil supply of NPs because, for the OTI bean cultivar, nano-priming did not affect germination; however, plant height, the total number of nodules, and seed K, and Zn concentrations were higher than those of the control (unprimed seeds). These results might be attractive to farmers, especially smallholder farmers in marginalized or rural areas with limited access to markets and healthcare systems and where diets are dominated by micronutrient-poor staple food [ 73 ]. The land dedicated to producing common beans is approximately 33 M hectares globally. In Africa, five million hectares of beans are cultivated by smallholder farmers, most of whom are women [ 65 ]. From an economic perspective, investing in nano-priming could be a cost-effective method to produce nutrient-rich food and a sustainable approach to combat hidden hunger. On the other hand, despite the increased costs of nano-priming compared to the control (unprimed), the benefits may be greater in the long term. For example, seed priming can extend the nodulation period without threatening plants´ vegetative growth while improving agronomic traits [ 16 ] or food quality. In that case, an opportunity is opened to lower the environmental impact of agriculture because global legume-rhizobial symbioses are estimated to fix 21 Mt of N annually, representing approximately one-tenth of ammonia applied annually synthesized by the Haber-Bosch environmentally expensive process. Moreover, scope two emissions, indirect greenhouse emissions associated with bean cropping, may be reduced, due to the global legume-rhizobial symbioses estimated to save CO 2 emissions of over 150 Mt annually [ 18 , 65 ]. Thus, nano-priming could be a complementary approach to producing beans, while enriching the soils via biological nitrogen fixation and enhancing the nutritional quality of bean grains. However, the effects of nano-priming on other bean varieties should be assessed to know their responses and to improve the nano-priming technology since some studies have reported that seed priming did not affect the germination of seeds. However it may burst in plant development throughout the plant´s life cycle [ 3 , 53 ], and these results agree with those observations. For instance, Acharya et al., [ 74 ] examined the effects of nano-priming with AgNPs on two watermelons ( Citrullus lanatus Riverside variety) seeds in three locations over three years (2017–2019). In 2017, no significant differences were observed in the days required for 50% seed germination, final germination percentage, and the emergence percentage between primed or unprimed seeds. However, in 2017, watermelon seed priming increased significantly the yield compared to that of the plants from unprimed seeds; meanwhile, the fruit quality was not modified. Despite the benefits of nano-priming, one of the drawbacks of the seed priming technique is that no standard protocol can be followed mindlessly [ 75 ]. It requires standardization before application to determine the appropriate “stop-time”, and re-drying [ 75 ] because these variables greatly depend on the plant species and properties of seeds. Thus, the lack of standardization of the priming technique prevents a convincing evaluation of the benefits of seed priming. Therefore, the main challenge in seed priming is standardization. Conclusions Nano-priming may be a complementary tool for improving the nutritional quality of beans. Encouraged results were obtained, although no external source of nutrients was added to bean plants. The hypothesis that nano-priming may improve germination traits compared to unprimed seeds and enhance the yield of bean plants could not be confirmed. Nano-priming with citrate-coated CoFe 2 O 4 NPs affected plant growth, nodulation, and mineral content of bean seeds. Nano-priming improved the nutritional quality of bean seeds in terms of Zn, and K concentration. A low phytic acid: Zn molar ratio was observed in the offspring seeds from the 10 mg L − 1 nano-priming treatment. In addition, the low cost of nano-priming of seeds may be a feasible approach to enhance the mineral content of beans and potentially contribute to the transition to a more sustainable agri-food system, as well as to enhance nutritional security in rural and marginalized areas because it is an easy and cheap tool. However, for its agronomical adoption and more homogenous plant response, standardization of the protocols is highly recommended for conventional seed priming and seed nano-priming. Declarations Author Contributions YSPV: Conceptualization, investigation, data curation, formal analysis, visualization, writing original draft; RCG: Conceptualization, methodology, resources, writing-review & editing; MCGC: Conceptualization, supervision, funding acquisition, resources, review & editing; JV: Validation, review & editing, supervision; DTM: Methodology and analysis with scanning electron microscopy, writing-review. JLL: review & editing. All the authors review and accept the final draft. Conflicts of interest There are no conflicts to declare. Acknowledgments The authors thank Dr. Serafin Cruz Izquierdo for providing the bean seeds, M.Sc. Jorge Valdez Carrasco for his help in the reflected light microscopy imaging. Funding This research received no external funding Data Availability Data generated or analyzed in this study are included in this paper and supplementary materials References Shanka D (2020) Roles of eco-friendly low input technologies in crop production in sub-Saharan Africa. Cogent Food Agric 6:1843882. https://doi.org/10.1080/23311932.2020.1843882 Vaidya S, Deng C, Wang Y, Zuverza-Mena N, Dimkpa C, White JC (2024) Nanotechnology in agriculture: A solution to global food insecurity in a changing climate? NanoImpact 34:100502. https://doi.org/10.1016/j.impact.2024.100502 do Espirito Santo Pereira A, Caixeta Oliveira H, Fernandes Fraceto L, Santaella C (2021) Nanotechnology potential in seed priming for sustainable agriculture. Nanomater. https://doi.org/10.3390/nano11020267 . Dutta P (2018) Seed priming: New vistas and contemporary perspectives. In: Rakshit A, and Singh HB (eds) Advances in seed priming. Springer, Singapore, pp. 3–22. https://doi.org/10.1007/978-981-13-0032-5_1 Nile SH, Thiruvengadam M, Wang Y, et al (2022) Nano-priming as emerging seed priming technology for sustainable agriculture—recent developments and future perspectives. J Nanobiotechnology 20:254. https://doi.org/10.1186/s12951-022-01423-8 Chandrasekaran U, Luo X, Wang Q, Shu K (2020) Are there unidentified factors involved in the germination of nanoprimed seeds? Front Plant Sci. 11:546690. https://doi.org/10.3389/fpls.2020.00832 Shah T, Latif S, Saeed F, Ali I, Ullah S, Abdullah Alsahli A, Jan S, Ahmad P (2021) Seed priming with titanium dioxide nanoparticles enhances seed vigor, leaf water status, and antioxidant enzyme activities in maize ( Zea mays L.) under salinity stress. J King Saud Univ - Sci 33:101207. https://doi.org/10.1016/j.jksus.2020.10.004 Sher A, Sarwar T, Nawaz A, Ijaz M, Sattar A, Ahmad S (2019) Methods of seed priming. In: Hasanuzzaman M, Fotopoulos V (eds) Priming and pretreatment of seeds and seedlings: implication in plant stress tolerance and enhancing productivity in crop plants. Springer Singapore, pp 1–10. https://doi.org/10.1007/978-981-13-8625-1_1 Shelar A, Singh A V, Maharjan RS, et al (2021) Sustainable agriculture through multidisciplinary seed nanopriming: prospects of opportunities and challenges. Cells 10:2428. https://doi.org/10.3390/cells10092428 Khan MN, Li Y, Khan Z, Chen L, Liu J, Hu J, Wu H, Li Z (2021) Nanoceria seed priming enhanced salt tolerance in rapeseed through modulating ROS homeostasis and α-amylase activities. J Nanobiotechnology 19:276. https://doi.org/10.1186/s12951-021-01026-9 Pagano A, Macovei A, Balestrazzi A (2023) Molecular dynamics of seed priming at the crossroads between basic and applied research. Plant Cell Rep 42:657–688. https://doi.org/10.1007/s00299-023-02988-w Rai-Kalal P, Jajoo A (2021) Priming with zinc oxide nanoparticles improve germination and photosynthetic performance in wheat. Plant Physiol Biochem 160:341–351. https://doi.org/10.1016/j.plaphy.2021.01.032 Abbasi Khalaki M, Moameri M, Asgari Lajayer B, Astatkie T (2021) Influence of nano-priming on seed germination and plant growth of forage and medicinal plants. Plant Growth Regul 93:13–28. https://doi.org/10.1007/s10725-020-00670-9 Sundaria N, Singh M, Upreti P, Chauhan RP, Jaiswal JP, Kumar A (2019) Seed priming with iron oxide nanoparticles triggers iron acquisition and biofortification in wheat ( Triticum aestivum L.) Grains. J Plant Growth Regul 38:122–131. https://doi.org/10.1007/s10725-020-00670-9 Ma J, Song Z, Yang J, Wang Y, Han H (2021) Cobalt ferrite nanozyme for efficient symbiotic nitrogen fixation via regulating reactive oxygen metabolism. Environ Sci Nano 8:188–203. http://dx.doi.org/10.1039/D0EN00935K Ma J, Zhou Y, Li J, Song Z, Han H (2022) Novel approach to enhance Bradyrhizobium diazoefficiens nodulation through continuous induction of ROS by manganese ferrite nanomaterials in soybean. J Nanobiotechnology 20:168. https://doi.org/10.1186/s12951-022-01372-2 Petry N, Boy E, Wirth JP, Hurrell RF (2015) Review: The potential of the common bean ( Phaseolus vulgaris ) as a vehicle for iron biofortification. Nutrients 7:1144–1173. https://doi.org/10.3390/nu7021144 Uebersax MA, Cichy KA, Gomez FE, Porch TG, Heitholt J, Osorno JM, Kamfwa K, Snapp SS, Bales S (2023) Dry beans ( Phaseolus vulgaris L.) as a vital component of sustainable agriculture and food security—A review. Legum Sci 5:e155. https://doi.org/10.1002/leg3.155 Murube E, Beleggia R, Pacetti D, et al (2021) Characterization of nutritional quality traits of a common bean germplasm collection. Foods 10:1572. https://doi.org/10.3390/foods10071572 Lisciani S, Marconi S, Le Donne C, et al (2024) Legumes and common beans in sustainable diets: nutritional quality, environmental benefits, spread and use in food preparations. Front Nutr 11:1385232. https://doi.org/10.3389/fnut.2024.1385232 De Souza-Torres A, Govea-Alcaide E, Gómez-Padilla E, Masunaga SH, Effenberger FB, Rossi LM, López-Sánchez R, Jardim RF (2021) Fe 3 O 4 nanoparticles and Rhizobium inoculation enhance nodulation, nitrogen fixation and growth of common bean plants grown in soil. Rhizosphere 17:100275. https://doi.org/10.1016/j.rhisph.2020.100275 Burridge JD, Findeis JL, Jochua CN, Miguel MA, Mubichi-Kut FM, Quinhentos ML, Xerinda SA, Lynch JP (2019) A case study on the efficacy of root phenotypic selection for edaphic stress tolerance in low-input agriculture: Common bean breeding in Mozambique. F Crop Res 244:107612. https://doi.org/10.1016/j.fcr.2019.107612 Estrada-Gómez JA, Estrada-Trejo V, Hernández-Livera A, Molina-Moreno JC, Campos-Escudero A (2004) OTI, una nueva variedad de frijol para el valle de México. Rev Fitotec Mex 27:115 Silva-Silva MJ, Mijangos-Ricardez OF, Vázquez-Hipólito V, Martinez-Vargas S, López-Luna J (2016) Single and mixed adsorption of Cd(II) and Cr(VI) onto citrate-coated magnetite nanoparticles. Desalin Water Treat 57:4008–4017. https://doi.org/10.1080/19443994.2014.991756 Martinez-Vargas S, Martínez AI, Hernández-Beteta EE, Mijangos-Ricardez OF, Vázquez-Hipólito V, Patiño-Carachure C, Hernandez-Flores H, López-Luna J (2017) Arsenic adsorption on cobalt and manganese ferrite nanoparticles. J Mater Sci 52:6205–6215. https://doi.org/10.1007/s10853-017-0852-9 Perea-Velez YS, González Chávez M del CA, Carrillo-González R, Lopez-Luna J (2022) Dissolution kinetics of citrate coated CoFe 2 O 4 nanoparticles in soil solution. Environ Sci Nano. 9:2954–2965. https://doi.org/10.1039/D2EN00330A Feizi H, Rezvani Moghaddam P, Shahtahmassebi N, Fotovat A (2012) Impact of bulk and nanosized titanium dioxide (TiO 2 ) on wheat seed germination and seedling growth. Biol Trace Elem Res 146:101–106. https://doi.org/10.1007/s12011-011-9222-7 Antony D, Yadav R, Kalimuthu R (2021) Accumulation of phyto-mediated nano-CeO 2 and selenium doped CeO 2 on Macrotyloma uniflorum (horse gram) seed by nano-priming to enhance seedling vigor. Biocatal Agric Biotechnol 31:101923. https://doi.org/10.1016/j.bcab.2021.101923 Del Buono D, Luzi F, Tolisano C, Puglia D, Di Michele A (2022) Synthesis of a lignin/zinc oxide hybrid nanoparticles system and its application by nano-priming in maize. Nanomater 12:568. https://doi.org/10.3390/nano12030568 Velu G, Bhattacharjee R, Rai K, Sahrawat K, Longvah T (2008) A simple and rapid screening method for grain zinc content in pearl millet. J SAT Agric Res 6:1–4 Ayala Garay AV, Acosta Gallegos JA, Reyes Muro L (2021) El cultivo del frijol presente y futuro para México. INIFAP, Guanajuato Hashem A, Abd_Allah EF, Alqarawi AA, Al-Huqail AA, Wirth S, Egamberdieva D (2016) The interaction between arbuscular mycorrhizal fungi and endophytic bacteria enhances plant growth of Acacia gerrardii under salt stress. Front Microbiol 7:1089. https://doi.org/10.3389/fmicb.2016.01089 Senthilkumar M, Amaresan N, Sankaranarayanan A (2021) Quantitative estimation of leghemoglobin content in legume root nodules. In: Plant-microbe interactions. Springer Protocols Handbooks. Springer, New York. pp 33–35. https://doi.org/10.1007/978-1-0716-1080-0_5 Kjeldahl J (1883) Neue methode zur bestimmung des stickstoffs in organischen körpern. Zeitschrift für Anal Chemie 22:366–382 Kitson RE, Mellon MG (1944) Colorimetric determination of phosphorus as molybdivanadophosphoric acid. Ind Eng Chem Anal Ed 16:379–383. https://doi.org/10.1021/i560130a017 Mariotti F, Tomé D, Mirand PP (2008) Converting nitrogen into protein—beyond 6.25 and Jones’ factors. Crit Rev Food Sci Nutr 48:177–184. https://doi.org/10.1080/10408390701279749 Gao Y, Shang C, Saghai Maroof MA, Biyashev RM, Grabau EA, Kwanyuen P, Burton JW, Buss GR (2007) A modified colorimetric method for phytic acid analysis in soybean. Crop Sci 47:1797–1803. https://doi.org/10.2135/cropsci2007.03.0122 Dhaliwal SS, Sharma V, Shukla AK, et al (2022) Biofortification of soybean ( Glycine max L.) through FeSO 4 *7H 2 O to enhance yield, iron nutrition and economic outcomes in sandy loam soils of India. Agriculture 47:1797–1803. https://doi.org/10.3390/agriculture12050586 Sarwar G, Hussain N, Schmeisky H, Muhammad S (2007) Use of compost an environment friendly technology for enhancing rice-wheat production in Pakistan. Pakistan J Bot 39:1553–1558 SADER (2022) Precios de garantía a productos alimentarios básicos, seguridad y certidumbre a productores. https://www.gob.mx/agricultura/articulos/precios-de-garantia-a-productos-alimentarios-basicos-seguridad-y-certidumbre-a-productores . Accessed 9 October 2022 Perea-Vélez YS, Carrillo-González R, González-Chávez M del CA, Vangronsveld J, Monasterio IO, Tapia Maruri D (2023) Citrate-coated cobalt ferrite nanoparticles for the nano-enabled biofortification of wheat. Food Funct. 14:4017–4035. https://doi.org/10.1039/D2FO03835H International Monetary Fund (2021) Representative exchange rates for selected currencies for september 2021. https://www.imf.org/external/np/fin/data/rms_mth.aspx?SelectDate=2021-09-30&reportType=REP . Accessed 9 October 2022 R Core Team (2020) R: A language and environment for statistical computing. Vienna, Austria, 2020. Romano A, Stevanato P (2020) Germination data analysis by time-to-event approaches. Plants. 9:617. https://doi.org/10.3390/plants9050617 Onofri A, Benincasa P, Mesgaran MB, Ritz C (2018) Hydrothermal-time-to-event models for seed germination. Eur J Agron 101:129–139. https://doi.org/10.1016/j.eja.2018.08.011 Onofri A, Mesgaran MB, Ritz C (2022) A unified framework for the analysis of germination, emergence, and other time-to-event data in weed science. Weed Sci 70:259–271. https://doi.org/10.1017/wsc.2022.8 Khan MN, Fu C, Li J, Tao Y, Li Y, Hu J, Chen L, Khan Z, Wu H, Li Z (2023) Seed nanopriming: How do nanomaterials improve seed tolerance to salinity and drought? Chemosphere 310:13691. https://doi.org/10.1016/j.chemosphere.2022.136911 Acharya A, Pal PK (2020) Agriculture nanotechnology: Translating research outcome to field applications by influencing environmental sustainability. NanoImpact 19:100232. https://doi.org/10.1016/j.impact.2020.100232 Savassa SM, Castillo-Michel H, Pradas del Real AE, Reyes-Herrera J, Marques JPR, de Carvalho HWP (2021) Ag nanoparticles enhancing Phaseolus vulgaris seedling development: understanding nanoparticle migration and chemical transformation across the seed coat. Environ Sci Nano 8:493–50. http://dx.doi.org/10.1039/D0EN00959H Savassa SM, Duran NM, Rodrigues ES, de Almeida E, van Gestel CAM, Bompadre TF V, P. de Carvalho HW (2018) Effects of ZnO nanoparticles on Phaseolus vulgaris germination and seedling development determined by X-ray spectroscopy. ACS Appl Nano Mater 1:6414–6426. https://doi.org/10.1021/acsanm.8b01619 Forest V, Cottier M, Pourchez J (2015) Electrostatic interactions favor the binding of positive nanoparticles on cells: A reductive theory. Nano Today 10:677–680. https://doi.org/10.1016/j.nantod.2015.07.002 Afzal S, Sharma D, Singh NK (2021) Eco-friendly synthesis of phytochemical-capped iron oxide nanoparticles as nano-priming agent for boosting seed germination in rice ( Oryza sativa L.). Environ Sci Pollut Res 28:40275–40287. https://doi.org/10.1007/s11356-020-12056-5 Ighaiee Oskoiee A, Ghanbari AA, Mirhadi MJ (2021) Effects of iron nanoparticles on seed germination of bean genotypes. SSRN Electron J. https://doi.org/10.2139/ssrn.3899301 Kandhol N, Singh VP, Ramawat N, Prasad R, Chauhan DK, Sharma S, Grillo R, Sahi S, Peralta-Videa J, Tripathi DK (2022) Nano-priming: Impression on the beginner of plant life. Plant Stress 5:100091. https://doi.org/10.1016/j.stress.2022.100091 Pawar VA, Ambekar JD, Kale BB, Apte SK, Laware SL (2019) Response in chickpea ( Cicer arietinum L.) seedling growth to seed priming with iron oxide nanoparticles. Int J Biosci 14:82–91. http://dx.doi.org/10.12692/ijb/14.3.82-91 Kabata-Pendias A (2010) Trace Elements in Soils and Plants (4th edition). CRC Press, Boca Raton. https://doi.org/10.1201/b10158 Plank CO, Kissel DE (1989) Plant Analysis Handbook for Georgia Rosenblueth M, Martínez-Romero E (2004) Rhizobium e tli maize populations and their competitiveness for root colonization. Arch Microbiol 181:337–344. https://doi.org/10.1007/s00203-004-0661-9 Zhang M-X, Zhao L-Y, He Y-Y, Hu J-P, Hu G-W, Zhu Y, Khan A, Xiong Y-C, Zhang J-L (2024) Potential roles of iron nanomaterials in enhancing growth and nitrogen fixation and modulating rhizomicrobiome in alfalfa ( Medicago sativa L.). Bioresour Technol 391:129987. https://doi.org/10.1016/j.biortech.2023.129987 Fedorova EE, Coba de la Peña T, Lara-Dampier V, Trifonova NA, Kulikova O, Pueyo JJ, Lucas MM (2021) Potassium content diminishes in infected cells of Medicago truncatula nodules due to the mislocation of channels MtAKT1 and MtSKOR/GORK. J Exp Bot 72:1336–1348. https://doi.org/10.1093/jxb/eraa508 Wang J, Cao X, Wang C, Chen F, Feng Y, Yue L, Wang Z, Xing B (2022) Fe-based nanomaterial-induced root nodulation is modulated by flavonoids to improve soybean ( Glycine max ) growth and quality. ACS Nano 16:21047–21062. https://doi.org/10.1021/acsnano.2c08753 Montes-Luz B, Conrado AC, Ellingsen JK, Monteiro RA, de Souza EM, Stacey G (2023) Acetylene reduction assay: A measure of nitrogenase activity in plants and bacteria. Curr Protoc. https://doi.org/10.1002/cpz1.766 Li M, Gao L, White JC, Haynes CL, O’Keefe TL, Rui Y, Ullah S, Guo Z, Lynch I, Zhang P (2023) Nano-enabled strategies to enhance biological nitrogen fixation. Nat Nanotechnol 18:688–69. https://doi.org/10.1038/s41565-023-01392-5 Chávez-Mendoza C, Sánchez E (2017) Bioactive compounds from mexican varieties of the common bean ( Phaseolus vulgaris ): Implications for health. Molecules 22:1360. https://doi.org/10.3390/molecules22081360 Huertas R, Karpinska B, Ngala S, et al (2022) Biofortification of common bean ( Phaseolus vulgaris L.) with iron and zinc: Achievements and challenges. Food Energy Secur 12:e406. https://doi.org/10.1002/fes3.406 Diaz S, Polania J, Ariza-Suarez D, Cajiao C, Grajales M, Raatz B, Beebe SE (2022) Genetic correlation between Fe and Zn biofortification and yield components in a common bean ( Phaseolus vulgaris L.). Front Plant Sci 12:739033. https://doi.org/10.3389/fpls.2021.739033 Beebe S (2020) Biofortification of common bean for higher iron concentration. Front Sustain Food Syst 4:573449. https://doi.org/10.3389/fsufs.2020.573449 Espinoza-García N, Martínez-Martínez R, Chávez-Servia JL, Vera-Guzmán AM, Carrillo-Rodríguez JC, Heredia-García E, Velasco-Velasco VA (2016) Contenido de minerales en semilla de poblaciones nativas de frijol común (Phaseolus vulgaris L.). Rev Fitotec Mex 39:215–223 HarvestPlus (2022) Hight iron beans. A food that can change your business and the world naturally. https://www.harvestplus.org/wp-content/uploads/2022/01/Iron-Beans.pdf . Accessed 9 October 2022 Hummel M, Talsma EF, Taleon V, Londoño L, Brychkova G, Gallego S, Raatz B, Spillane C (2020) Iron, zinc and phytic acid retention of biofortified, low phytic acid, and conventional bean varieties when preparing common household recipes. Nutrients 12:658. https://doi.org/10.3390/nu12030658 Castro-Alba V, Lazarte CE, Bergenståhl B, Granfeldt Y (2019) Phytate, iron, zinc, and calcium content of common Bolivian foods and their estimated mineral bioavailability. Food Sci Nutr 7:2854–2865. https://doi.org/10.1002/fsn3.1127 Siimes N, Sharp EL, Lewis N, Kah M (2022) Determining acceptance and rejection of nano-enabled agriculture: A case study of the New Zealand wine industry. NanoImpact 28:100432. https://doi.org/10.1016/j.impact.2022.100432 Garg M, Sharma N, Sharma S, Kapoor P, Kumar A, Chunduri V, Arora P (2018) Biofortified crops generated by breeding, agronomy, and transgenic approaches are improving lives of millions of people around the world. Front Nutr 5:12. https://doi.org/10.3389/fnut.2018.00012 Acharya P, Jayaprakasha GK, Crosby KM, Jifon JL, Patil BS (2020) Nanoparticle-mediated seed priming improves germination, growth, yield, and quality of watermelons ( Citrullus lanatus ) at multi-locations in Texas. Sci Rep 10:5037. https://doi.org/10.1038/s41598-020-61696-7 Paul S, Dey S, Kundu R (2022) Seed priming: an emerging tool towards sustainable agriculture. Plant Growth Regul 97:215–234. https://doi.org/10.1007/s10725-021-00761-1 Additional Declarations No competing interests reported. Supplementary Files FigS1.docx TableS1.docx TableS2.docx TableS3.docx TableS4.docx TableS5.docx TableS6.docx Cite Share Download PDF Status: Published Journal Publication published 30 Aug, 2024 Read the published version in Journal of Nanoparticle Research → Version 1 posted Editorial decision: Revision requested 09 Jul, 2024 Reviews received at journal 09 Jul, 2024 Reviewers agreed at journal 26 Jun, 2024 Reviewers invited by journal 20 Jun, 2024 Editor assigned by journal 19 Jun, 2024 Submission checks completed at journal 18 Jun, 2024 First submitted to journal 13 Jun, 2024 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 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-4578599","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":321139934,"identity":"24761aae-eaae-48f5-bf30-cc32bdae59ec","order_by":0,"name":"Yazmín Stefani Perea-Vélez","email":"","orcid":"","institution":"Colegio de Postgraduados","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yazmín","middleName":"Stefani","lastName":"Perea-Vélez","suffix":""},{"id":321139936,"identity":"f79b8634-511c-4626-ba0b-4c99b1c945f2","order_by":1,"name":"Rogelio Carrillo-González","email":"","orcid":"","institution":"Colegio de Postgraduados","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Rogelio","middleName":"","lastName":"Carrillo-González","suffix":""},{"id":321139938,"identity":"5067d646-2e91-4540-8140-6a3e84935f1e","order_by":2,"name":"Ma. Carmen A. González-Chávez","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAw0lEQVRIiWNgGAWjYBADOQYG5gbStBgzMDCSqCWxgWgt/O1nn274uKM2vb+9se3Bhz8M0QYHCGiROJNudnPmmeO5M84cbDec2caQO5OQZQYMaWy3eduO5W6QSGyT5m1gyO0n5DAD/mdst/+2HUs3AGnh+cOQ20ZQiwTQFsa2mgSIFjYibJG48YztZm/bAUOoXyQI+4W/P43txs+2Onn+9uZjwBCzyd1wgJA1EHAYRLCBbCVOPRDUwbSMglEwCkbBKMAEAGfnQzX+Uw0PAAAAAElFTkSuQmCC","orcid":"","institution":"Colegio de Postgraduados","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Ma.","middleName":"Carmen A.","lastName":"González-Chávez","suffix":""},{"id":321139940,"identity":"340e4656-4066-4403-b858-1d2ac5744b8f","order_by":3,"name":"Jaco Vangronsveld","email":"","orcid":"","institution":"Hasselt University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jaco","middleName":"","lastName":"Vangronsveld","suffix":""},{"id":321139942,"identity":"2e477b0b-509a-4676-8537-97a793878481","order_by":4,"name":"Daniel Tapia-Maruri","email":"","orcid":"","institution":"Instituto Politécnico Nacional","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Daniel","middleName":"","lastName":"Tapia-Maruri","suffix":""},{"id":321139944,"identity":"d1b6adf7-c44e-467c-ab79-c1327e6b11be","order_by":5,"name":"Jaime López-Luna","email":"","orcid":"","institution":"Instituto de Ecología, Red de Estudios Moleculares Avanzados","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jaime","middleName":"","lastName":"López-Luna","suffix":""}],"badges":[],"createdAt":"2024-06-14 00:32:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4578599/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4578599/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11051-024-06101-4","type":"published","date":"2024-08-30T15:57:26+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":59680825,"identity":"4f330b14-dbaa-4659-aaf0-640e73950237","added_by":"auto","created_at":"2024-07-04 18:04:54","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":12474345,"visible":true,"origin":"","legend":"\u003cp\u003eLocalization of Fe on seed coat and cotyledons by Perls’ Prussian blue staining. Pictures a) and b) show the blue staining among the seed coats from the unprimed and primed seeds. Picture c) shows variations in staining intensity among the cotyledons. The white arrows show the staining due to the NPs. SC, seed coat; H, hilum; M, micropyle.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/3d974233caef6b1802581d73.png"},{"id":59680601,"identity":"e1ba3565-0c73-475e-aaab-920db3562abe","added_by":"auto","created_at":"2024-07-04 17:56:54","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":693097,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of nano-priming on germination of OTI beans. a) Germination curve, b) fitted parametric log-logistic model time-to-event, and c) no parametric model time-to-event. For the time-to-event curves, the markers represent the observed data, and the lines show the time-to-event model fit.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/a8d0443e6118e6f3f012e254.png"},{"id":59680603,"identity":"f8130aea-5e52-4741-9d56-d92b21f2458b","added_by":"auto","created_at":"2024-07-04 17:56:54","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":402357,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of nano-priming on the emergence and plant growth of OTI bean. a) Emergence curve, and b) growth curve. The dotted line represents the Max, maximum, Avg, average, and Min, minimum plant height reported in the varietal description. * Symbol indicates that the simple main effect of nano-priming treatment was significant according to the repeated measures analysis. Different letters represent significant differences between growth curves according to the Bonferroni p-adjusted method.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/060620f7a3fa8660512c784d.png"},{"id":63821341,"identity":"7a17834f-7d97-421c-8949-12a00ea3e495","added_by":"auto","created_at":"2024-09-02 16:13:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":24243895,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/b958a613-88a4-4ddc-8881-d0276af13e85.pdf"},{"id":59680267,"identity":"6f4f95c4-9c8a-4943-9d15-941f35045eea","added_by":"auto","created_at":"2024-07-04 17:48:53","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":58291,"visible":true,"origin":"","legend":"","description":"","filename":"FigS1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/e7710dc6774a11d961aeba27.docx"},{"id":59680600,"identity":"5ecbc5b8-dede-475b-a34a-3385c0dbde06","added_by":"auto","created_at":"2024-07-04 17:56:53","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":16787,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/793daf566a3b248fa21c9619.docx"},{"id":59680269,"identity":"f452e6de-78bf-4d1b-aaa6-64bec66de1ca","added_by":"auto","created_at":"2024-07-04 17:48:54","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":17401,"visible":true,"origin":"","legend":"","description":"","filename":"TableS2.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/0c8034649c002605c9ecbdc1.docx"},{"id":59680274,"identity":"4c45098f-546f-4999-94ac-82c9d7a9f15b","added_by":"auto","created_at":"2024-07-04 17:48:54","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":20371,"visible":true,"origin":"","legend":"","description":"","filename":"TableS3.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/d78f2a2cabb325be636376dc.docx"},{"id":59680271,"identity":"3b06d93e-353c-4310-86e2-d42cc78a0969","added_by":"auto","created_at":"2024-07-04 17:48:54","extension":"docx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":16883,"visible":true,"origin":"","legend":"","description":"","filename":"TableS4.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/46f4cb8dc4a84c7680fee7eb.docx"},{"id":59680276,"identity":"bf85a2f5-fcad-4f9c-b584-af1f7966a3cb","added_by":"auto","created_at":"2024-07-04 17:48:54","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":17379,"visible":true,"origin":"","legend":"","description":"","filename":"TableS5.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/a42babdd0a76cd8a6f0200d5.docx"},{"id":59680272,"identity":"4b5d0f17-6370-4ff3-9eec-d4a65be3baea","added_by":"auto","created_at":"2024-07-04 17:48:54","extension":"docx","order_by":7,"title":"","display":"","copyAsset":false,"role":"supplement","size":20610,"visible":true,"origin":"","legend":"","description":"","filename":"TableS6.docx","url":"https://assets-eu.researchsquare.com/files/rs-4578599/v1/d71186ad789a89787bb8a17c.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Nano-priming of Phaseolus vulgaris OTI cultivar with cobalt ferrite nanoparticles enhances the mineral composition of progeny seeds","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAdopting a sustainable model that is profitable, productive, and conserves natural resources is a challenge for agriculture today, and the soils are one of the most relevant elements in this new agricultural model. The premise is that it is no longer enough to maintain its quality, but it is also necessary to be able to improve it. In this regard, low-input technologies (such as seed priming, cover crops/green manure, or intercrops) can be one of the several possible ways to achieve sustainability [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. On the one hand, nano-enabled agriculture has shown that through nano-fertilizers it is possible to improve plant health, nutrient biofortification, increased photosynthetic output, and higher rates of nitrogen fixation[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Among the nano-enabled technologies, seed priming or nano-priming is an emerging field proven to be more promising than traditional priming approaches [\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSeed priming is a low-input pre-sowing seed treatment. It allows synchronized germination and maturity of crops, improves germination time and water use efficiency, and increases nutrient uptake and tolerance of plants to biotic and abiotic stresses [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Seed priming is generally defined as the controlled hydration of seeds to a level that allows pre-germinative metabolic activity without radical protrusions [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. After hydration, the seed is dried again to its original weight [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Unlike traditional seed priming, which employs water or solutions containing nutrients, hormones, microorganisms, or biopolymers [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e], nano-priming uses nanoparticle (NPs) suspensions or nano-formulations [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Numerous nanomaterials have been tested as priming agents, for instance, Ag, Au, Cu, TiO\u003csub\u003e2\u003c/sub\u003e, FeS\u003csub\u003e2\u003c/sub\u003e, Fe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e, and ZnO [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan additionalcitationids=\"CR11 CR12\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], proving that they can significantly increase the germination rate, vigor index, root elongation, plumule length, and the antioxidant response of seedling of many crops [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. However, ferrite NPs (such as CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, NiFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4,\u003c/sub\u003e and Fe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) have attracted attention for nano-enabled agriculture due to their biocompatibility, and safety for humans and plants. For instance, Sundaria et al. [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e] reported increased germination percentage and shoot length in two wheat varieties (high and low iron efficiency) after nano-priming with Fe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs. Cobalt ferrite (CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eNPs can significantly enhance the symbiotic nitrogen fixation efficiency by 260% [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], and increase the root nodulation by 61% in \u003cem\u003eGlycine max\u003c/em\u003e (L.) Merr. compared to the control plants [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOn the other hand, \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e (common bean) was selected for this study because is a protein source for more than 300\u0026nbsp;million people from Latin America, the Caribbean, and Eastern Africa [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. It is considered a nearly perfect food because of its high protein, fiber, prebiotic, vitamin (A, C, and folate), and mineral content (Ca, Mg, K, Cu, Fe, and Zn) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. For this reason, beans are a tool for fighting to hide hunger. Another interesting aspect to consider is that the common bean is a crop that enriches the soil through biological nitrogen fixation, which is based on symbiosis with bacteria such as \u003cem\u003eRhizobium leguminosarium bv. Phaseoli\u003c/em\u003e [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The use of leguminous as intercrop in arable systems can reduce nitrous oxide emission by 18%, and N fertilizer use by 24% compared to systems without legumes [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Although the common bean is considered an inefficient N fixer compared to other legumes [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], bean plants are used as an intercrop in Africa by smallholder farmers.\u003c/p\u003e \u003cp\u003eIn this context, this study aimed to evaluate the effects of nano-priming on the life cycle of \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L. OTI cultivar with citrate-coated cobalt ferrite (CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) NPs. We hypothesized that (1) nano-priming may improve germination traits compared to unprimed seeds, (2) enhance yield, and (3) improve the nutritional quality of bean grains. Moreover, the cost of the nano-priming was estimated. The results of this research can help to provide accessible solutions to smallholder farmers, because the low productivity of these farmers is mainly related to the limited use of inputs, such as irrigation, pesticides, mechanization, and fertilizers [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. In this regard, nano-priming can be an alternative to reduce the dependency on external fertilizer purchases. On the other hand, the use of NPs to increase the nodulation, and N fixation capacity of beans is a breakthrough attempt at sustainable agriculture.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cp\u003eTwo experiments were conducted to assess the effect of nano-priming with citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs on the life cycle of \u003cem\u003eP. vulgaris\u003c/em\u003e OTI cultivar. The first experiment evaluated the effect of nano-priming on the germination stage, while the second experiment focused on its effects from the vegetative to maturity stages of bean plants. For quality assurance and quality control in analytical measurements, the material was washed in a 0.25 M HCl solution to remove any traces of contaminating materials. All the reagents used were of analytical grade (J.T. Baker, Merck, and Sigma\u0026ndash;Aldrich), and standard solutions were prepared using certified stock solutions.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant material\u003c/h2\u003e \u003cp\u003e \u003cem\u003eP. vulgaris\u003c/em\u003e OTI cultivar seeds were obtained from the Genetic Resources and Productivity Program of Colegio de Postgraduados. OTI is an improved bean cultivar that is adapted to the central high valleys of the Mexican Republic. The main characteristics of OTI beans are the cooking time (61\u0026ndash;85 min), its yield (2.5 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), and its high resistance to \u003cem\u003eColletotrichum lindemuthianum\u003c/em\u003e, \u003cem\u003eSclerotinia\u003c/em\u003e sp., and \u003cem\u003eRhizoctonia solani\u003c/em\u003e; medium resistance to \u003cem\u003eUromyces appendiculathus\u003c/em\u003e var. appendiculatus and moderate tolerance to \u003cem\u003ePseudomonas phaceolicola\u003c/em\u003e. The seed coat of OTI beans has a light brown background with bright purple spots [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eSynthesis and properties of citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs\u003c/h2\u003e \u003cp\u003eCitrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs were synthesized using a co-precipitation method. Eight mL of 1 M Fe(NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e3\u003c/sub\u003e and 4 mL of 1M Co(NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e solutions were mixed. Then, 38 mL of 1.5 M NaOH solution was added dropwise to the metal ion solution under constant stirring conditions (400 rpm). The mixture solution was continuously stirred and heated at 90\u0026deg;C for 1h. The resulting black precipitate was washed three times with deionized water. The precipitate was then mixed with 50 mL sodium citrate 4 mM solution and stirred at 80\u0026deg;C for 30 min. After that, the NPs were washed with deionized water until the pH of the supernatant was 7 [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. The washed NPs were suspended in 50 mL of deionized water to prepare a NPs stock solution. The NPs characterization data were published by Perea-V\u0026eacute;lez et al. (2022), with results indicating that the NPs had a semi-spherical shape with an average primary size of 13\u0026thinsp;\u0026plusmn;\u0026thinsp;5 nm, hydrodynamic diameter of 216\u0026thinsp;\u0026plusmn;\u0026thinsp;10 nm, zeta potential in deionized water of 10.5\u0026thinsp;\u0026plusmn;\u0026thinsp;6.6 mV, point of zero charge of 6.8, and 48% Fe, 29% Co composition.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eExperiment one. Assessment of the effect of nano-priming on the germination of bean\u003c/h2\u003e \u003cp\u003eThe seed water uptake curve was evaluated before the start of the experiment (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). This determined the imbibition time for nano-priming because each crop cultivar has its critical soaking duration, and the success of priming depends on achieving optimal seed hydration. So, according to this assay, the soaking time was fixed at 2 h and 30 min. Afterward, a completely randomized experimental design was set up to assess the effect of nano-priming on the germination traits of the \u003cem\u003eP. vulgaris\u003c/em\u003e OTI cultivar. The treatment factor was the concentration of the NPs suspension with three levels: 10, 20, and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (equivalent to 3.5, 7.5, and 15 mg Fe L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, and 2.4, 4.5. and 9.5 mg Co L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, respectively). The concentrations of the nano-priming suspensions were chosen to ensure the dispersion of NPs and to avoid their fast agglomeration. Furthermore, to differentiate between water and NPs effects, seeds were primed with distilled water (hydro-priming or positive control), and unprimed seeds were used as a negative control. Three replicates were used for each treatment (clear-hinged containers with ten seeds).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of priming suspension and nano-priming\u003c/h2\u003e \u003cp\u003eThe priming suspensions were freshly prepared in sterilized deionized water before their use. An aliquot of NPs stock solution was placed in a Flask to get a NPs concentration of 10, 20, and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The NPs suspension was sonicated with an ultrasonic probe for 2 min at 130 W and 90% amplitude. On the other hand, bean seeds were surface-sterilized by soaking them in a 3% (v/v) sodium hypochlorite solution for 10 min and then carefully rinsed with sterilized deionized water to remove all chlorine. Afterward, the seeds were soaked in the NPs suspension for 2 h 30 min at room temperature (20\u0026ndash;25\u0026deg;C) with constant agitation on a rotor shaker (30 turns per minute). For the hydro-priming, distilled water was used instead of NPs suspension. The primed seeds were then dried back to their original moisture content; for this, the seeds were placed in sterilized paper bags and dried in an oven at 25\u0026deg;C for 3 d. Dried seeds were then stored at 4\u0026deg;C for fifteen days until further use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eSeed germination assay\u003c/h2\u003e \u003cp\u003eTen seeds were placed in a clear-hinged plastic container that was surface-disinfected. The bottom of the container was lined with sterilized cotton and two sterile filter papers. The seeds were moistened with 15 mL of sterile distilled water and kept in the darkness at 24\u0026deg;C for 7 d to initiate germination. Germinating seeds were observed and counted daily based on radical emergence up to 2 mm [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Germination percentage (GP), energy period (EP), germination energy (GE), germination rate (GR), and mean germination time (MGT) were calculated based on the following equations [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]:\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$GP=\\frac{number of seeds germinated}{total number of seeds placed for germination test}\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equb\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equb\" name=\"EquationSource\"\u003e\n$$EP=Days after starting the germination test to reach \\ge 50\\text{%} of germinated seeds$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\n$$GE=\\frac{Cumulative daily total of germinated seeds at day 4}{Number of seeds set to germinate}\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equd\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equd\" name=\"EquationSource\"\u003e\n$$GR=\\left(\\frac{a}{1}\\right)+\\left(\\frac{b-a}{2}\\right)+\\left(\\frac{c-b}{3}\\right)+\\dots +\\left(\\frac{n-{n}_{-1}}{D}\\right)$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Eque\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Eque\" name=\"EquationSource\"\u003e\n$$mean germination time \\left(MGT\\right)=\\frac{\\sum \\left(D\\times n\\right)}{\\sum n}$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003ewhere a, b, c, \u0026hellip;, n is the number of seeds germinated on days 1, 2, 3, \u0026hellip;, D, and D is the number of days counted from the beginning of the test.\u003c/p\u003e \u003cp\u003eThe fresh and dry weights of the seedlings were determined at the end of the germination experiment (day seven). Stem height and root length were measured using the ImageJ software. The germination index (GI) and the relative seed germination (RSG) were calculated according to the following equations [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]:\u003cdiv id=\"Equf\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equf\" name=\"EquationSource\"\u003e\n$$GI=(G\\times L)/(Gw\\times Lw)$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equg\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equg\" name=\"EquationSource\"\u003e\n$$RSG=\\frac{Seeds germinated from nanopriming }{Seeds germinated from hydro-priming or control }\\times 100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eWhere G and L are the germination and radicle length recorded for a specific treatment, respectively, and Gw and Lw are the values recorded for the control seeds.\u003c/p\u003e \u003cp\u003eThe seedling vigor was calculated based on the following equation [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]:\u003cdiv id=\"Equh\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equh\" name=\"EquationSource\"\u003e\n$$Vigor index =germination \\text{%} \\times seedling weight (root+shoot)$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eQuantitative estimation of Fe and Co content in nano-primed seeds\u003c/h2\u003e \u003cp\u003eGround dried primed seeds (0.5 g) were acid-digested (1 mL H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e and 4 mL HClO\u003csub\u003e4\u003c/sub\u003e:H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e; 4:1 v/v), and the digested sample was diluted to 25 mL with deionized water. The Fe and Co concentrations were determined using flame atomic absorption spectroscopy (Perkin Elmer, model 3110).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eLocalization of NPs and iron in nano-primed seeds\u003c/h2\u003e \u003cp\u003eThe Perls\u0026rsquo; Prussian blue staining technique was employed for the initial detection of NPs within seeds [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. This process involved placing dried primed seeds in distilled water for one hour to hydrate the primed seeds partially. Subsequently, the primed seeds were incubated with the Prussian blue solution for one hour to allow staining. After staining, the seeds were thoroughly rinsed with distilled water, and excess water was removed using paper tissue. The samples were then mounted on microscope slides and subjected to qualitative analysis using a reflectance light microscope (Carl Zeiss Stereo V20). High-resolution images were captured by using a Canon 5D digital camera.\u003c/p\u003e \u003cp\u003eEnvironmental scanning electron microscopy ESEM analysis was performed to confirm the presence of the NPs in the seeds. The dried nano-primed seeds were mounted on double-sided carbon conductive tape and observed using an ESEM (Carl Zeiss EVO LS10, Jena, Germany) microscope and an X-ray detector (Bruker, Quantax 200, Germany). Images were taken at 30 kV and 80 Pa of water vapor pressure.\u003c/p\u003e \u003cp\u003e \u003cb\u003eExperiment two. Effect of nano-priming on the growth of bean plants (from VE to R2 growth stage) and nodulation (greenhouse experiment)\u003c/b\u003e \u003c/p\u003e \u003cp\u003eA completely randomized experimental design was used in this study. The treatments were as follows: negative control (unprimed seeds), positive control (hydro-primed seeds), and nano-primed seeds at 10, 20, and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. Each treatment had ten replicates.\u003c/p\u003e \u003cp\u003eClay soil collected from the surface layer of the experimental field of Colegio de Postgraduados (19\u0026deg;27\u0026rsquo;56.0\u0026rdquo; N, 98\u0026deg;54\u0026rsquo; 10.4\u0026rdquo; W) was used in this experiment. The soil properties are listed in Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e. The soil was air-dried for at least one week in the shadow. Then, it was sieved to \u0026lt;\u0026thinsp;2 mm and manually homogenized with a shovel. No external source of macro or micronutrients was added to the soil because soil nutrient content was high enough to fulfill the nutrient demands of beans (53 kg N t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 8 kg P t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 55 kg K t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 40 kg Ca t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 8 kg Mg t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 271 g Fe t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 90 g Cu t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, and 192 g Zn t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) according to Ayala Garay et al. [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTwo primed or unprimed seeds were sown in pots containing 8 kg of soil. Seedling emergence was recorded when the cotyledons were completely raised above the soil. At the V2 growth stage (when the second trifoliate leaf was established), one of the seedlings was removed. The plant height was recorded once a week after plant emergence. The plants were grown in an open greenhouse from April to June 2022 at an average temperature of 21\u0026deg;C (maximum and minimum temperatures of 27\u0026deg;C and 14\u0026deg;C, respectively). Plants were watered daily with tap water to maintain approximately 60% of the soil field moisture capacity.\u003c/p\u003e \u003cp\u003eTo assess the effect of nano-priming on nodulation, four plants per treatment were carefully harvested 51 d after sowing (R2 plant growth stage), and shoots were separated from the roots. Shoots were used to determine the leaf area and plant nutrition, while fresh nodules were collected, and the nodule fresh weight and number of active nodules (pink pigmented) or inactive nodules (brown dark-green) per plant root were determined.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eNitrogenase activity\u003c/h2\u003e \u003cp\u003eNitrogenase activity was determined by acetylene reduction. For plants at the R2 growth stage (51 d after sowing), samples of roots with nodules were transferred into a plastic hermetic closed container (1.5 L), and the excess adhered soil was carefully removed before placing the roots in the container. A serum stopper was inserted, and 10% of the air was replaced with acetylene and incubated for 1 h [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. The gas samples were analyzed using a gas chromatograph (Clarus 400, PerkinElmer).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eFoliar area\u003c/h2\u003e \u003cp\u003eDigital images (Moto g6 camera) of leaves arranged on a white background and under bright light conditions were taken. Images were analyzed using the ImageJ analysis software.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eEvaluation of plant nutrition\u003c/h2\u003e \u003cp\u003eThe aerial part of the plant was dried in an oven at 70\u0026deg;C for 3 d and then ground into a powder. An aliquot of 500 mg was acid-digested (one mL H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e and four mL HClO\u003csub\u003e4\u003c/sub\u003e: H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e, 4:1 v/v) in a digest block for 16 h at 90\u0026deg;C. The digested samples were then made up to 25 mL with deionized water and filtered. The Ca, Mg, Fe, and Zn concentrations were determined using flame atomic absorption spectroscopy (Perkin Elmer, model 3110). The N concentration was calculated by the Kjeldahl method [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e], while the P and K concentration was determined using the vanadomolybdo phosphoric acid colorimetric method [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] and flame photometry (Jenway, PFP7), respectively.\u003c/p\u003e \u003cp\u003e \u003cb\u003eAssessment of the effects of nano-priming on the agronomic traits and grain nutritional quality of bean plants\u003c/b\u003e \u003c/p\u003e \u003cp\u003ePlants (6) were harvested when they reached maturity (R9, 81 d after sowing). The agronomic variables evaluated were the number of pods per plant, weight of pods, number of seeds per pod and plant, seed yield (weight of seeds per plant), seed index (100 seed weight), and harvest index.\u003c/p\u003e \u003cp\u003eThe macro (Ca, Mg, K, and P) and micronutrient concentrations (Fe, Zn, and Cu) in the seeds were determined by atomic absorption spectroscopy (Perkin Elmer, model 3110) or flame photometry (Jenway PFP7). The P concentration in the seeds was determined using the vanamolybdo phosphoric acid colorimetric method [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. The protein concentration in the seeds was evaluated using the Kjeldahl method [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. The protein content was calculated by multiplying the amount of total nitrogen by a conversion factor of 6.25, which assumes that the nitrogen content of proteins in foodstuffs is 16% [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The phytic acid concentration in seeds was estimated using the colorimetric Wade reagent method, modified as described by Gao et al. [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. The molar ratios of phytic acid to Fe and Zn were calculated by converting the concentrations of Fe (55.84 g mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), Zn (65.38 g mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), and phytic acid (660.04 g mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) into moles.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eEconomic evaluation of nano-priming seed treatment\u003c/h2\u003e \u003cp\u003eThe economic analysis was performed using the economic indicators described by Dhaliwal et al. [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e] and Sarwar et al. [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]:\u003cdiv id=\"Equi\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equi\" name=\"EquationSource\"\u003e\n$$Gross income=yield \\left(t {ha}^{-1}\\right)\\times grain price$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equj\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equj\" name=\"EquationSource\"\u003e\n$$Profitable return \\left(PR\\right)=gross income-total production cost$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equk\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equk\" name=\"EquationSource\"\u003e\n$$PR over control=PR-control treatment$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equl\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equl\" name=\"EquationSource\"\u003e\n$$Cost benefit ratio \\left(CBR\\right)=\\frac{PR over control}{Total production cost}$$\u003c/div\u003e\u003c/div\u003e\u003cdiv id=\"Equm\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equm\" name=\"EquationSource\"\u003e\n$$Investment factor \\left(IF\\right)=\\frac{Gross income}{Total production cost}$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eThe estimation of the gross income indicator relied on the established guaranteed price of beans in Mexico in 2022 (MXN 16, 000 t\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) as reported by Mexico\u0026rsquo;s Secretariat of Agriculture and Rural Development [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. To calculate production costs, data from the five leading bean-producing states in Mexico were obtained from the Trust Funds for Rural Development (FIRA in Spanish; Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). The cost of the nano-priming treatment was determined by considering the price of nanoparticles, which amounted to 5 750 USD per kg [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. To report the economic indicators in terms of USD, the exchange rate used was 19.9 MXN/USD [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e], which was an average of the exchange rates between 1 and 15 June 2022.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eData analysis\u003c/h2\u003e \u003cp\u003eThe data were analyzed with the statistical software R version 4.0.3 [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. Germination data were analyzed using a time-to-event model [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e] because this type of analysis provides more reasonable inferences considering that the germination event did not occur at the specific time of evaluation but during the interval between assessments. Parametric and non-parametric approaches were applied for data analysis. The goodness-of-fit model was evaluated graphically from the observed versus the predicted values. In the case of the parametric time-to-event model, a likelihood ratio test was performed to compare the time-to-event models between the different treatments. Model optimization was done using the Akaike Information Criterion (AIC), where the selected model had the lowest AIC. In the case of no parametric approach to modeling the effect of the experimental factor, a Wilcoxon-type statistic was carried out [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. Plant growth (plant height) data were analyzed using repeated-measures ANOVA. Subsequently, pairwise comparisons between time points at each and between group levels were performed using the Bonferroni p-adjustment method. On the other hand, an analysis of variance was used (α\u0026thinsp;=\u0026thinsp;0.05) to compare the effect of seed priming on germination traits, agronomic traits, and seed nutritional quality. Before ANOVA analysis, Shapiro\u0026rsquo;s and Bartlett\u0026rsquo;s tests were used to verify the compliance with the assumptions of normality and homogeneity of variances. Data that did not exhibit assumptions of normality and homogeneity of variance (GP, GR, EP, GE, MGT, relative seed germination vs. control treatment, number of nodules, and fresh weight of nodules) were analyzed using the Kruskal-Wallis rank sum test. The data for phytic acid and P grain concentrations were box cox-transformed. The Tukey honest significant difference test (α\u0026thinsp;=\u0026thinsp;0.05) and Bonferroni test were used to detect differences between treatments.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and discussion","content":"\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eNano-priming of OTI bean and its effects on the germination variables\u003c/h2\u003e \u003cp\u003eThe seed Fe concentration, after nano-priming with 10, 20, and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, were 53.31\u0026thinsp;\u0026plusmn;\u0026thinsp;3.58, 53.72\u0026thinsp;\u0026plusmn;\u0026thinsp;6.31, and 57.27\u0026thinsp;\u0026plusmn;\u0026thinsp;2.99 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, respectively. The seed Fe concentration was not significantly (p\u0026thinsp;=\u0026thinsp;0.0571) different from that of unprimed (60.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22) and hydro-primed (53.73\u0026thinsp;\u0026plusmn;\u0026thinsp;2.99) seeds. Cobalt was not detected in the acid digestion extracts of the primed seeds. After nano-priming, the NPs may or may not be taken up by the seeds, but It has been suggested that most NPs may remain on the seed surface after nano-priming [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Histochemical staining and microscopic analyses were used to confirm the presence and location of NPs on or within the seed. These analyses complemented the atomic absorption analysis, which did not reveal significant differences in Fe seed concentration between primed and unprimed seeds. Based on the staining intensity, the NPs were localized in the hilum (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb white arrows) and seed coat (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea white arrows). In comparison, the blue staining intensity of cotyledons increased with increasing NPs concentration (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec). Elemental mapping analysis also confirmed the presence of Fe and Co in the cotyledons (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). These findings suggest that NPs adhere to the seed coat and are taken up by the seed through the hilum region. This is consistent with previous studies that showed that Ag, Cu, Zn, and Fe NPs remained within the common bean seed coat, mainly in the hilum area.[\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e] The adhesion and uptake of NPs may be attributed to their physical and chemical properties, such as size, charge, and surface chemistry [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. The positive surface charge (zeta potential 10.5\u0026thinsp;\u0026plusmn;\u0026thinsp;6.6 mV [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] of the citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs favors the electrostatics interaction of NPs with the negatively charged groups on the seed coat surface [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. However, the precise mechanism involved in the interaction between NPs and the seed coat requires further investigation to optimize the use of NPs for seed priming applications, as this aspect has received relatively little attention so far [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe seed water uptake after nano-priming ranged from 40%\u0026plusmn;3\u0026ndash;48%\u0026plusmn;4%, but no significant changes (p\u0026thinsp;=\u0026thinsp;0.101) were observed compared to the hydro-priming (52.3%\u0026plusmn; 5.2%). These results contrast with other studies where nano-priming promoted faster water uptake compared to conventional seed priming techniques. For example, Afzal et al. [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e] reported 50% more water uptake in rice grains imbibed with 20 and 40 mg FeO NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for 24 h compared with hydro-priming and FeSO\u003csub\u003e4\u003c/sub\u003e solution (20 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). Similar results were observed with other types of NPs as priming agents and other seeds. For instance, nano-priming of rapeseeds (\u003cem\u003eBrassica napus\u003c/em\u003e variety Zhongshuang 11) with polyacrylic acid-coated nanoceria (0.1 mM) increased the water uptake by 52% compared to the priming with TES buffer in the first hour of the priming. The increment of water uptake in nano-primed seeds after 3 and 8 h of imbibition was 14% and 12% more compared to the observed with TES buffer treatment [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. In wheat grains, nano-priming with ZnO NPs (10 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) increased water uptake by 51% compared to hydro-priming after 12 h of imbibition [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. It has been pointed out that nano-priming accelerates the seed water uptake. However, the mechanisms of water uptake into the seed under the influence of NM are still unknown [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. Moreover, the effects of NPs on plant or seed performance depend on the properties and concentrations of NPs, plant species, and seed properties [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eUsing traditional evaluation methods, which consider variables such as GP, GI, RSG, EP, MGT, and GR, it was observed that nano-priming did not exert a significant influence on the germination of OTI beans compared to hydro-priming and control treatments (Table \u003cspan refid=\"MOESM3\" class=\"InternalRef\"\u003eS3\u003c/span\u003e). However, the time-to-event analysis revealed differences in the germination curves between treatments. Non-parametric time-to-event curves (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec) showed that the germination of control seeds and those nano-primed with 10 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e differed from the other treatments, as confirmed by Wilcoxon scores. The sum of the Wilcoxon scores for control and nano-primed seeds at 10 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e were 7.57 and 3.57, respectively, while negative values were observed for hydro-primed seeds (-2.08), and nano-primed seeds at 20 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-2.54) and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (-6.52). Thus, the germination of control and nano-primed seeds with 10 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e was, on average, the fastest compared to the other treatments. This conclusion was further supported by the parametric time-to-event curves, which estimated the lowest median germination time for control and nano-primed seeds at 10 mg of NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Overall, the parametric approach of time-to-event models is well-suited for making inferences about underlying biological mechanisms, such as germination [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. However, the likelihood ratio test for the parametric log-logic time-to-event model curves (p\u0026thinsp;=\u0026thinsp;0.0164) indicated that the germination curves were different between treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb). Based on the AIC criterion, the control treatment had the fastest germination, followed by 10 mg NPs, hydro-priming, 40 mg NPs, and 20 mg NPs. The nano-priming did not affect the fresh (p\u0026thinsp;=\u0026thinsp;0.066) and dry weight (p\u0026thinsp;=\u0026thinsp;0.176) of seedlings, average root length (p\u0026thinsp;=\u0026thinsp;0.068), root diameter (p\u0026thinsp;=\u0026thinsp;0.195), or the number of roots (p\u0026thinsp;=\u0026thinsp;0.313; Table \u003cspan refid=\"MOESM4\" class=\"InternalRef\"\u003eS4\u003c/span\u003e). No significant differences (p\u0026thinsp;=\u0026thinsp;0.061) were found between the vigor index of seedlings from the nano-priming and control (Table \u003cspan refid=\"MOESM4\" class=\"InternalRef\"\u003eS4\u003c/span\u003e). These observations contrast with studies that stated that nano-priming accelerates or enhances germination and improves seedling growth and vigor in different plant species [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. In rice, nano-priming with FeO NPs at 20 and 40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e increased radicle length by 50% and plumule length by 22% compared with hydro-priming [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. Pawar et al. [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e] found that Fe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e NPs at 4\u0026ndash;8 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e significantly enhanced the radicle length and plumule length of chickpea (\u003cem\u003eCicer arietinum\u003c/em\u003e L variety Digvijay) seedlings.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eParameter estimates from the parametric time-to-event model and the Akaike information criterion (AIC) for the germination of OTI beans after priming treatment.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eParameter\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHydro-priming\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10 mg NPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20 mg NPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e40 mg NPs\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSlope\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27.40 (29.26)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11.77* (2.19)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e14.67* (2.58)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8.21* (1.52)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e12.37* (2.35)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGF\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.97* (0.03)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.93* (0.05)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.97* (0.03)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.94* (0.05)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.90* (0.05)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMGT\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.01* (0.13)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.39* (0.11)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.17* (0.09)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.44* (0.15)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.59* (0.12)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e90.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e106.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e94.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e123.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e108.45\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003e\u0026dagger;\u003c/sup\u003eGF, germinate fraction; \u003csup\u003e\u0026dagger;\u003c/sup\u003eMGT, mean germination time for the germinate fraction; * indicates that the estimate is significant; data in parentheses are standard errors of estimates.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eEffect of nano-priming on the life cycle of OTI bean plants and the agronomic traits\u003c/h2\u003e \u003cp\u003eFrom primed seeds, seedling emergence between 80% and 100% was observed 8 d after sowing (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). One hundred % of seedling emergence was recorded 11 d after sowing. Overall, the plants (from unprimed and primed seeds) had a shorter growing duration (81\u0026ndash;83 d after sowing) than the varietal description (110\u0026ndash;130 d after sowing), and flowering occurred 11 d before the expected time (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003eThe plant height (until R2, flowering) resulting from primed seeds exceeded the maximum height reported in the varietal description (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). The simple main effect of nano-priming was significant at 14 d (p\u0026thinsp;=\u0026thinsp;0.018) and 20 d (p\u0026thinsp;=\u0026thinsp;0.013) for treatments with NPs at 10 and 20 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). When comparing the plant growth curves (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb), plants from treated seeds with NPs at 20 and 40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e were taller (75.1\u0026thinsp;\u0026plusmn;\u0026thinsp;13.0, and 68.9\u0026thinsp;\u0026plusmn;\u0026thinsp;12.7 cm, respectively) than those of the control (43.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.1 cm) and nano-priming with 10 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (61.8\u0026thinsp;\u0026plusmn;\u0026thinsp;20.0 cm). On the other hand, in terms of the vegetative growth parameters (foliar area, fresh and dry weight of shoot and root system; Table \u003cspan refid=\"MOESM5\" class=\"InternalRef\"\u003eS5\u003c/span\u003e), plant performance until the flowering stage was not affected by the nano-priming compared to the control or hydro-priming treatment (p\u0026thinsp;\u0026gt;\u0026thinsp;0.1).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe nutritional status of plants at the flowering stage (R2, 51 d after sowing) was within the normal values of reference for macro-and micronutrients for soybean and other beans (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), except for N, Ca, Mg, and Fe, the concentrations of these elements was higher than the typical values found for beans. When comparing the plants from the nano-priming with those from the control and hydro-priming, no significant differences were observed in the concentrations of macronutrients (N, P, K, Ca, and Mg), Fe, and Cu. Nevertheless, the Zn concentration in the leaves of plants treated with 20 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e was higher than in plants treated with 10 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of NPs.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eNutritional status of OTI bean plants at the flowering stage (R2, 51 d after sowing) and reference values.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eK\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMg\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c10\" namest=\"c8\"\u003e \u003cp\u003emg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28.6\u0026thinsp;\u0026plusmn;\u0026thinsp;5.4 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e47.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e487.3\u0026thinsp;\u0026plusmn;\u0026thinsp;152 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e15.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHydro-priming\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e47.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e418.5\u0026thinsp;\u0026plusmn;\u0026thinsp;95 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e11.4\u0026thinsp;\u0026plusmn;\u0026thinsp;7.0 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10 mg NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28.5\u0026thinsp;\u0026plusmn;\u0026thinsp;3.7 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e45.1\u0026thinsp;\u0026plusmn;\u0026thinsp;3.6 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e472.3\u0026thinsp;\u0026plusmn;\u0026thinsp;125 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e10.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20 mg NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e25.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e57.9\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e305.7\u0026thinsp;\u0026plusmn;\u0026thinsp;230 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e12.4\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e40 mg NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e47.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.0 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e510.6\u0026thinsp;\u0026plusmn;\u0026thinsp;160 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e8.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"10\" nameend=\"c10\" namest=\"c1\"\u003e \u003cp\u003eReference values\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSufficient or normal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.25\u0026ndash;6.0\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.30\u0026ndash;6.00\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;2.0\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.8\u0026ndash;3.0\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.30\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.0\u0026ndash;150.0\u003csup\u003eδ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e25.0\u0026ndash;300.0\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e5.0\u0026ndash;30.0 \u003csup\u003eδ\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation of n\u0026thinsp;=\u0026thinsp;4. Different letters represent significant differences between the treatments (Tukey\u0026rsquo;s α\u0026thinsp;=\u0026thinsp;0.05). and ns means that no significant differences were detected between the treatments. Reference values for interpreting plant analyses were obtained according to \u003csup\u003eδ\u003c/sup\u003e Kabata-Pendias [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e] and \u003csup\u003e\u0026dagger;\u003c/sup\u003ePlank and Kissel [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eRegarding nodule formation at the flowering stage (51d after sowing), no significant differences were found in the number of active nodules and their fresh weights due to the treatment factor. It is worth mentioning that the observed nodules were products of native rhizobia present in the soil. The soil used has a history of growing maize plants, and symbiotic bacteria, such as \u003cem\u003eRhizobium etli\u003c/em\u003e, are naturally associated with the rhizosphere of maize and inside their roots [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. In contrast, the reduction of acetylene (to ethylene C\u003csub\u003e2\u003c/sub\u003eH\u003csub\u003e4\u003c/sub\u003e) was only detected in the roots of plants from the 40 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e nano-primed seeds. The produced C\u003csub\u003e2\u003c/sub\u003eH\u003csub\u003e4\u003c/sub\u003e was 343.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1 \u0026micro;mol h\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, this result is in agreement with De Souza-Torres et al. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e] who found that the application of iron oxide (Fe\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) NPs, through irrigation water, stimulated the nitrogenase activity (from 4.3 to 6.5 \u0026micro;mol h\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and nodulation of common bean plants compared to control plants (without NPs supplementation). Zhang et al. [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e] reported that seed soaking plus leaf spraying of 10 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of FeNPs significantly increased the nitrogenase activity by 91% compared with the control plants of alfalfa (45 d old after sowing). On the other hand, at the R9 stage (maturation), it was observed that plants derived from nano-primed seeds with 40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e exhibited the highest number of total nodules (187\u0026thinsp;\u0026plusmn;\u0026thinsp;35) compared to both the control plants (40\u0026thinsp;\u0026plusmn;\u0026thinsp;25) and those subjected to hydro-priming (84\u0026thinsp;\u0026plusmn;\u0026thinsp;55, Table \u003cspan refid=\"MOESM6\" class=\"InternalRef\"\u003eS6\u003c/span\u003e). The maximum change in the total nodules between flowering and plant maturity stages was observed in plants from the nano-priming seeds with 20 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (3.38-fold change), followed by plants from the nano-priming with 40 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (2.74-fold change), hydro-priming (1.47-fold change), nano-priming with 10 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (0.36-fold change) and control treatment (0.25-fold change). However, future analysis should show that the nodules after flowering are active during this period because the root nodules have a short life span (12\u0026ndash;18 d) [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e], and there was a lapse of 28 days between flowering and harvesting.\u003c/p\u003e \u003cp\u003eThere is limited information regarding the effect of nano-priming with CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs on biological nitrogen fixation, but our observations agree with other studies that yielded promising results regarding the effects of Fe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and Fe\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs on root nodulation. For instance, Ma et. al. [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] found that the supplement of CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e at 10 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in the growth medium (river sand) enlarged and increased total nodules by ~\u0026thinsp;30%, in soybean plants (\u003cem\u003eGlycine max\u003c/em\u003e L. Meer \u0026ldquo;Williams 82\u0026rdquo;) compared to the control treatment (without amendment of NPs). Ma et al. [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] demonstrated that MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs enhanced nodulation in soybeans, leading to a 61% increase in nodules and a 51% increase in nodule weight compared to the control (without NPs), along with improved nitrogen fixation (~\u0026thinsp;2 to 2.5 times higher than the control). Wang et al. [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e] showed a 35% increase in root nodules in soybean through foliar application of Fe\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs compared with the unexposed plants. Comparisons between studies are challenging due to varying experimental conditions, crops, and NPs application methods, these findings underscore the potential positive impact of NPs on biological nitrogen fixation and nodulation. Increasing the capacity of biological nitrogen fixation can reduce or even eliminate the need for synthetic fertilizer [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e] and is an effective strategy to enhance food security [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e]. In this regard, nanotechnology and nano-priming may offer a pathway to improve biological nitrogen fixation, but a more comprehensive understanding of plant-bacterial-NPs interaction is essential.\u003c/p\u003e \u003cp\u003eFor the agronomic traits (Table \u003cspan refid=\"MOESM6\" class=\"InternalRef\"\u003eS6\u003c/span\u003e), nano-priming and hydro-priming did not have significant effects on plant dry weight (p\u0026thinsp;=\u0026thinsp;0.947), stem diameter (p\u0026thinsp;=\u0026thinsp;0.754), number of pods (p\u0026thinsp;=\u0026thinsp;0.448), seed yield per plant (p\u0026thinsp;=\u0026thinsp;0.997), seed index (p\u0026thinsp;=\u0026thinsp;0.0.692), and harvest index (p\u0026thinsp;=\u0026thinsp;0.536). However, pod size, number of seeds per pod, and seed index (weight of 100 seeds) were within the values reported in the varietal description.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eEffect of nano-priming on the nutritional quality of OTI bean seeds\u003c/h2\u003e \u003cp\u003eTheir high protein and mineral contents characterize beans compared to cereals.[\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e] In the present study, the protein content in all treatments ranged from 106\u0026ndash;137%. Despite the lack of significant differences between offspring seeds from the primed seeds (nano-and hydro-primed), the control, protein concentration was higher than the reported protein content in Mexican bean varieties (14% and 33%) [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNano-priming affected the mineral composition of bean seeds, as shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Progeny seeds from plants grown from nano-primed seeds with 20 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, exhibited higher K concentrations than those from the control treatment. Unexpectedly, the highest Zn seed concentration was observed in the offspring seeds from plants of primed seeds with 10 and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e compared to the control, hydro-primed. The Zn concentration in the seeds was higher than the value range reported for Mexican bean varieties (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), as well as the average Zn concentration in beans (28\u0026ndash;31 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) from other regions around the globe [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e, \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. The Zn concentration in beans from the prime treatments was 29% higher than the extreme values observed in beans, such as 77 mg Zn kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e, \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. The target value for Zn biofortification is 17 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e above the local materials in each country or region [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e]. Thus, offspring from the primed seeds can be considered biofortified, as the Zn concentration was between 32 and 59 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e above the average Zn concentration for Mexican bean varieties. Additionally, the Zn concentration in progeny seeds from the 10 and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e primed seeds compared favorably with those found in animal products, where the Zn concentration ranged between 23 and 170 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e on a dry weight basis [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. In contrast, no significant effects of the nano-priming treatment on the Ca, Mg, Fe, and Cu concentrations were observed in the offspring seeds. Nevertheless, the Fe seed concentrations among the treatments were higher than the average Fe concentration reported for seeds of Mexican bean varieties (35\u0026ndash;58 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e, \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. Meanwhile, it was below the target concentration for biofortification (140 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). According to the CIAT, the target Fe concentration for beans must be at least 94 mg Fe kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e above the concentration of local varieties in each country or region [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe potential bioavailability of Fe and Zn depends on their concentration of anti-nutritional compounds, like phytic acid (PA), polyphenols, lectins, and tannins. Among those compounds, it is suggested that PA is one of the primary and significant inhibitors of mineral bioavailability (and thus uptake) from beans, followed by polyphenols [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e]. The PA concentrations ranged from 0.35 to 0.40 g 100 g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for the progeny seeds of plants grown from nano-primed seeds. The PA concentration observed in the control, and primed treatments was statistically similar. However, the PA concentration of tested seeds was below the average phytic acid concentration reported for Mexican bean varieties and bean seeds around the world (from 0.4 to 2.6 g 100 g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The relative bioavailability of Fe and Zn was determined based on the phytic acid to mineral molar ratios. In the case of Fe, the PA: Fe molar ratio was 5:1 on average, and no significant effect was found in the nano-priming treatment. Likewise, these PA: Fe ratios were lower than the range (from 6:1 to 33:1) observed in different bean seeds worldwide [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. For Zn, the lowest ratio (3:1) was observed in seeds from the plants from primed 10 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e seeds. Both values can be interpreted as low bioavailability compared to the recommended values (1:1) for adequate bioavailability [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMineral composition, phytic acid concentration, and phytic acid: iron or zinc (phy:Fe, phy:Zn) molar ratio of OTI bean seeds from plants of primed or unprimed seeds.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eReference values\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHydro-priming\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10 mg NPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20 mg NPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e40 mg NPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMexican varieties*\u003csup\u003e, δ\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eHarvestPlust\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMacronutrients (%)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.11\u0026ndash;0.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.11\u0026ndash;0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eK\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14 abc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.85\u0026ndash;0.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.27\u0026ndash;0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e106.89\u0026thinsp;\u0026plusmn;\u0026thinsp;14.2 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e135.26\u0026thinsp;\u0026plusmn;\u0026thinsp;39.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e127.39\u0026thinsp;\u0026plusmn;\u0026thinsp;12.3 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e136.68\u0026thinsp;\u0026plusmn;\u0026thinsp;18.8 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e133.20\u0026thinsp;\u0026plusmn;\u0026thinsp;18.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14\u0026ndash;33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e190\u0026ndash;230\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMicronutrients (mg kg\u003c/b\u003e\u003csup\u003e\u003cb\u003e\u0026minus;\u0026thinsp;1\u003c/b\u003e\u003c/sup\u003e\u003cb\u003e)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFe\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67.23\u0026thinsp;\u0026plusmn;\u0026thinsp;1.90 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e70.71\u0026thinsp;\u0026plusmn;\u0026thinsp;4.11 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e62.91\u0026thinsp;\u0026plusmn;\u0026thinsp;2.07 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e68.13\u0026thinsp;\u0026plusmn;\u0026thinsp;7.07 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e74.91\u0026thinsp;\u0026plusmn;\u0026thinsp;9.17ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e35\u0026ndash;58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e64\u0026ndash;119\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eZn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e78.9\u0026thinsp;\u0026plusmn;\u0026thinsp;14.3 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e77.8\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e84.1\u0026thinsp;\u0026plusmn;\u0026thinsp;6.8 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e82.8\u0026thinsp;\u0026plusmn;\u0026thinsp;7.6 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e27\u0026ndash;41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.14\u0026ndash;2.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOthers\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhytic acid (g 100g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.98\u0026ndash;2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhy:Fe ratio\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.96\u0026thinsp;\u0026plusmn;\u0026thinsp;1.54 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.10\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.81 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePhy:Zn ratio\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.97 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.86 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation of n=. Different letters represent significant differences between the treatments (Tukey\u0026rsquo;s α\u0026thinsp;=\u0026thinsp;0.05), and ns means that no significant differences were detected between the treatments. Phy:Fe ratio, phytic and iron molar ratio, Phy:Zn ratio, phytic and zinc molar ratio. Reference values of the mineral composition of bean seeds for Mexican varieties according to * Ch\u0026aacute;vez-Mendoza and S\u0026aacute;nchez [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e], and \u003csup\u003eδ\u003c/sup\u003e Espinoza-Garc\u0026iacute;a et al. [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. \u003csup\u003e\u0026dagger;\u003c/sup\u003eReference values for protein and Fe reported for the bean varieties of the HarvestPlus program [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eCost, benefits, and challenges of nano-priming seed treatment\u003c/h2\u003e \u003cp\u003eThe gross income estimated for the seed priming (including hydro- and nano-priming) was from 3,139.75 to 3,590.79 USD per ha (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Meanwhile, the gross income from unprimed seeds was 3,521.79 USD. No significant differences (p\u0026thinsp;=\u0026thinsp;0.83) were observed in the gross income between the control and the nano-priming. The profitable return of nano-priming over the control (unprimed seeds) was between 163.08 and 524.70 USD less. The negative profitable return of nano-priming over the unprimed control can be explained by the relatively low (but not statically significant) seed yield (Table \u003cspan refid=\"MOESM6\" class=\"InternalRef\"\u003eS6\u003c/span\u003e) of plants from the nano-primed seeds. However, seed priming may be a commercially viable form of incorporating nanotechnology into agriculture compared with the soil application or foliar spray of nano-fertilizer of micronutrients because seed priming is an inexpensive and straightforward method to enhance crop quality in resource-limited areas. For example, the calculated cost of nano-priming bean seeds (with citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs) ranged from 121 to 143 USD per ha and used a concentration suspension from 10 to 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. For wheat fortification, the estimated cost ranged from 44,283 to 65,523 USD per ha for soil fertilization (98\u0026ndash;145 mg CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), and 1,553 USD for foliar fertilization per ha [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEconomic indicators for the economic evaluation of the nano-priming seed treatment of OTI bean per hectare of cultivated land.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGross income (USD)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCost of production (USD)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eProfitable return (USD)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eProfitable return over control (USD)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eBenefit-cost ratio\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eInvestment factor\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3,521.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e936.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2,584.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3.76\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHydro-priming\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3,590.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e937.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2,653.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e68.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3.83\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10 mg NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3,479.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1,057.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2,421.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e-163.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3.29\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e20 mg NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3,421.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1,065.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2,356.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e-228.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3.21\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e40 mg NPs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3,139.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1,079.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2,060.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e-524.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e-0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e2.91\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe potential cost of nanotechnology is a crucial concern for farmers and one of the main factors considered for investing in this new technology [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. Farmers from the wine industry of New Zealand expressed acceptance of this novel technology if the nanotechnology can significantly reduce the cost of production, shorten production time, or enhance quality without increasing cost. Indeed, they also expressed that they are willing to prioritize quality over profitability [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. As mentioned above, nano-priming is a cheaper approach for implementing NPs in agriculture compared to the foliar or soil supply of NPs because, for the OTI bean cultivar, nano-priming did not affect germination; however, plant height, the total number of nodules, and seed K, and Zn concentrations were higher than those of the control (unprimed seeds). These results might be attractive to farmers, especially smallholder farmers in marginalized or rural areas with limited access to markets and healthcare systems and where diets are dominated by micronutrient-poor staple food [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e]. The land dedicated to producing common beans is approximately 33 M hectares globally. In Africa, five million hectares of beans are cultivated by smallholder farmers, most of whom are women [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. From an economic perspective, investing in nano-priming could be a cost-effective method to produce nutrient-rich food and a sustainable approach to combat hidden hunger.\u003c/p\u003e \u003cp\u003eOn the other hand, despite the increased costs of nano-priming compared to the control (unprimed), the benefits may be greater in the long term. For example, seed priming can extend the nodulation period without threatening plants\u0026acute; vegetative growth while improving agronomic traits [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] or food quality. In that case, an opportunity is opened to lower the environmental impact of agriculture because global legume-rhizobial symbioses are estimated to fix 21 Mt of N annually, representing approximately one-tenth of ammonia applied annually synthesized by the Haber-Bosch environmentally expensive process. Moreover, scope two emissions, indirect greenhouse emissions associated with bean cropping, may be reduced, due to the global legume-rhizobial symbioses estimated to save CO\u003csub\u003e2\u003c/sub\u003e emissions of over 150 Mt annually [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. Thus, nano-priming could be a complementary approach to producing beans, while enriching the soils via biological nitrogen fixation and enhancing the nutritional quality of bean grains. However, the effects of nano-priming on other bean varieties should be assessed to know their responses and to improve the nano-priming technology since some studies have reported that seed priming did not affect the germination of seeds. However it may burst in plant development throughout the plant\u0026acute;s life cycle [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e], and these results agree with those observations. For instance, Acharya et al., [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e] examined the effects of nano-priming with AgNPs on two watermelons (\u003cem\u003eCitrullus lanatus\u003c/em\u003e Riverside variety) seeds in three locations over three years (2017\u0026ndash;2019). In 2017, no significant differences were observed in the days required for 50% seed germination, final germination percentage, and the emergence percentage between primed or unprimed seeds. However, in 2017, watermelon seed priming increased significantly the yield compared to that of the plants from unprimed seeds; meanwhile, the fruit quality was not modified.\u003c/p\u003e \u003cp\u003eDespite the benefits of nano-priming, one of the drawbacks of the seed priming technique is that no standard protocol can be followed mindlessly [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e]. It requires standardization before application to determine the appropriate \u0026ldquo;stop-time\u0026rdquo;, and re-drying [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e] because these variables greatly depend on the plant species and properties of seeds. Thus, the lack of standardization of the priming technique prevents a convincing evaluation of the benefits of seed priming. Therefore, the main challenge in seed priming is standardization.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eNano-priming may be a complementary tool for improving the nutritional quality of beans. Encouraged results were obtained, although no external source of nutrients was added to bean plants. The hypothesis that nano-priming may improve germination traits compared to unprimed seeds and enhance the yield of bean plants could not be confirmed. Nano-priming with citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs affected plant growth, nodulation, and mineral content of bean seeds. Nano-priming improved the nutritional quality of bean seeds in terms of Zn, and K concentration. A low phytic acid: Zn molar ratio was observed in the offspring seeds from the 10 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e nano-priming treatment. In addition, the low cost of nano-priming of seeds may be a feasible approach to enhance the mineral content of beans and potentially contribute to the transition to a more sustainable agri-food system, as well as to enhance nutritional security in rural and marginalized areas because it is an easy and cheap tool. However, for its agronomical adoption and more homogenous plant response, standardization of the protocols is highly recommended for conventional seed priming and seed nano-priming.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eYSPV: Conceptualization, investigation, data curation, formal analysis, visualization, writing original draft; RCG: Conceptualization, methodology, resources, writing-review \u0026amp; editing; MCGC: Conceptualization, supervision, funding acquisition, resources, review \u0026amp; editing; JV: Validation, review \u0026amp; editing, supervision; DTM: Methodology and analysis with scanning electron microscopy, writing-review. JLL: review \u0026amp; editing. All the authors review and accept the final draft.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are no conflicts to declare.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors thank Dr. Serafin Cruz Izquierdo for providing the bean seeds, M.Sc. Jorge Valdez Carrasco for his help in the reflected light microscopy imaging.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research received no external funding\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData generated or analyzed in this study are included in this paper and supplementary materials\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eShanka D (2020) Roles of eco-friendly low input technologies in crop production in sub-Saharan Africa. Cogent Food Agric 6:1843882. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/23311932.2020.1843882\u003c/span\u003e\u003cspan address=\"10.1080/23311932.2020.1843882\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVaidya S, Deng C, Wang Y, Zuverza-Mena N, Dimkpa C, White JC (2024) Nanotechnology in agriculture: A solution to global food insecurity in a changing climate? NanoImpact 34:100502. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.impact.2024.100502\u003c/span\u003e\u003cspan address=\"10.1016/j.impact.2024.100502\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003edo Espirito Santo Pereira A, Caixeta Oliveira H, Fernandes Fraceto L, Santaella C (2021) Nanotechnology potential in seed priming for sustainable agriculture. Nanomater. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/nano11020267\u003c/span\u003e\u003cspan address=\"10.3390/nano11020267\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDutta P (2018) Seed priming: New vistas and contemporary perspectives. In: Rakshit A, and Singh HB (eds) Advances in seed priming. Springer, Singapore, pp. 3\u0026ndash;22. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-981-13-0032-5_1\u003c/span\u003e\u003cspan address=\"10.1007/978-981-13-0032-5_1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNile SH, Thiruvengadam M, Wang Y, et al (2022) Nano-priming as emerging seed priming technology for sustainable agriculture\u0026mdash;recent developments and future perspectives. J Nanobiotechnology 20:254. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12951-022-01423-8\u003c/span\u003e\u003cspan address=\"10.1186/s12951-022-01423-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChandrasekaran U, Luo X, Wang Q, Shu K (2020) Are there unidentified factors involved in the germination of nanoprimed seeds? Front Plant Sci. 11:546690. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2020.00832\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2020.00832\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShah T, Latif S, Saeed F, Ali I, Ullah S, Abdullah Alsahli A, Jan S, Ahmad P (2021) Seed priming with titanium dioxide nanoparticles enhances seed vigor, leaf water status, and antioxidant enzyme activities in maize (\u003cem\u003eZea mays\u003c/em\u003e L.) under salinity stress. J King Saud Univ - Sci 33:101207. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jksus.2020.10.004\u003c/span\u003e\u003cspan address=\"10.1016/j.jksus.2020.10.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSher A, Sarwar T, Nawaz A, Ijaz M, Sattar A, Ahmad S (2019) Methods of seed priming. In: Hasanuzzaman M, Fotopoulos V (eds) Priming and pretreatment of seeds and seedlings: implication in plant stress tolerance and enhancing productivity in crop plants. Springer Singapore, pp 1\u0026ndash;10. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-981-13-8625-1_1\u003c/span\u003e\u003cspan address=\"10.1007/978-981-13-8625-1_1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShelar A, Singh A V, Maharjan RS, et al (2021) Sustainable agriculture through multidisciplinary seed nanopriming: prospects of opportunities and challenges. Cells 10:2428. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/cells10092428\u003c/span\u003e\u003cspan address=\"10.3390/cells10092428\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhan MN, Li Y, Khan Z, Chen L, Liu J, Hu J, Wu H, Li Z (2021) Nanoceria seed priming enhanced salt tolerance in rapeseed through modulating ROS homeostasis and α-amylase activities. J Nanobiotechnology 19:276. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12951-021-01026-9\u003c/span\u003e\u003cspan address=\"10.1186/s12951-021-01026-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePagano A, Macovei A, Balestrazzi A (2023) Molecular dynamics of seed priming at the crossroads between basic and applied research. Plant Cell Rep 42:657\u0026ndash;688. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00299-023-02988-w\u003c/span\u003e\u003cspan address=\"10.1007/s00299-023-02988-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRai-Kalal P, Jajoo A (2021) Priming with zinc oxide nanoparticles improve germination and photosynthetic performance in wheat. Plant Physiol Biochem 160:341\u0026ndash;351. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.plaphy.2021.01.032\u003c/span\u003e\u003cspan address=\"10.1016/j.plaphy.2021.01.032\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbbasi Khalaki M, Moameri M, Asgari Lajayer B, Astatkie T (2021) Influence of nano-priming on seed germination and plant growth of forage and medicinal plants. Plant Growth Regul 93:13\u0026ndash;28. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10725-020-00670-9\u003c/span\u003e\u003cspan address=\"10.1007/s10725-020-00670-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSundaria N, Singh M, Upreti P, Chauhan RP, Jaiswal JP, Kumar A (2019) Seed priming with iron oxide nanoparticles triggers iron acquisition and biofortification in wheat (\u003cem\u003eTriticum aestivum\u003c/em\u003e L.) Grains. J Plant Growth Regul 38:122\u0026ndash;131. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10725-020-00670-9\u003c/span\u003e\u003cspan address=\"10.1007/s10725-020-00670-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMa J, Song Z, Yang J, Wang Y, Han H (2021) Cobalt ferrite nanozyme for efficient symbiotic nitrogen fixation via regulating reactive oxygen metabolism. Environ Sci Nano 8:188\u0026ndash;203. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1039/D0EN00935K\u003c/span\u003e\u003cspan address=\"10.1039/D0EN00935K\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMa J, Zhou Y, Li J, Song Z, Han H (2022) Novel approach to enhance \u003cem\u003eBradyrhizobium diazoefficiens\u003c/em\u003e nodulation through continuous induction of ROS by manganese ferrite nanomaterials in soybean. J Nanobiotechnology 20:168. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12951-022-01372-2\u003c/span\u003e\u003cspan address=\"10.1186/s12951-022-01372-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePetry N, Boy E, Wirth JP, Hurrell RF (2015) Review: The potential of the common bean (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e) as a vehicle for iron biofortification. Nutrients 7:1144\u0026ndash;1173. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/nu7021144\u003c/span\u003e\u003cspan address=\"10.3390/nu7021144\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUebersax MA, Cichy KA, Gomez FE, Porch TG, Heitholt J, Osorno JM, Kamfwa K, Snapp SS, Bales S (2023) Dry beans (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L.) as a vital component of sustainable agriculture and food security\u0026mdash;A review. Legum Sci 5:e155. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/leg3.155\u003c/span\u003e\u003cspan address=\"10.1002/leg3.155\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMurube E, Beleggia R, Pacetti D, et al (2021) Characterization of nutritional quality traits of a common bean germplasm collection. Foods 10:1572. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/foods10071572\u003c/span\u003e\u003cspan address=\"10.3390/foods10071572\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLisciani S, Marconi S, Le Donne C, et al (2024) Legumes and common beans in sustainable diets: nutritional quality, environmental benefits, spread and use in food preparations. Front Nutr 11:1385232. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fnut.2024.1385232\u003c/span\u003e\u003cspan address=\"10.3389/fnut.2024.1385232\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Souza-Torres A, Govea-Alcaide E, G\u0026oacute;mez-Padilla E, Masunaga SH, Effenberger FB, Rossi LM, L\u0026oacute;pez-S\u0026aacute;nchez R, Jardim RF (2021) Fe\u003csub\u003e3\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles and \u003cem\u003eRhizobium\u003c/em\u003e inoculation enhance nodulation, nitrogen fixation and growth of common bean plants grown in soil. Rhizosphere 17:100275. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.rhisph.2020.100275\u003c/span\u003e\u003cspan address=\"10.1016/j.rhisph.2020.100275\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBurridge JD, Findeis JL, Jochua CN, Miguel MA, Mubichi-Kut FM, Quinhentos ML, Xerinda SA, Lynch JP (2019) A case study on the efficacy of root phenotypic selection for edaphic stress tolerance in low-input agriculture: Common bean breeding in Mozambique. F Crop Res 244:107612. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fcr.2019.107612\u003c/span\u003e\u003cspan address=\"10.1016/j.fcr.2019.107612\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEstrada-G\u0026oacute;mez JA, Estrada-Trejo V, Hern\u0026aacute;ndez-Livera A, Molina-Moreno JC, Campos-Escudero A (2004) OTI, una nueva variedad de frijol para el valle de M\u0026eacute;xico. Rev Fitotec Mex 27:115\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSilva-Silva MJ, Mijangos-Ricardez OF, V\u0026aacute;zquez-Hip\u0026oacute;lito V, Martinez-Vargas S, L\u0026oacute;pez-Luna J (2016) Single and mixed adsorption of Cd(II) and Cr(VI) onto citrate-coated magnetite nanoparticles. Desalin Water Treat 57:4008\u0026ndash;4017. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/19443994.2014.991756\u003c/span\u003e\u003cspan address=\"10.1080/19443994.2014.991756\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMartinez-Vargas S, Mart\u0026iacute;nez AI, Hern\u0026aacute;ndez-Beteta EE, Mijangos-Ricardez OF, V\u0026aacute;zquez-Hip\u0026oacute;lito V, Pati\u0026ntilde;o-Carachure C, Hernandez-Flores H, L\u0026oacute;pez-Luna J (2017) Arsenic adsorption on cobalt and manganese ferrite nanoparticles. J Mater Sci 52:6205\u0026ndash;6215. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10853-017-0852-9\u003c/span\u003e\u003cspan address=\"10.1007/s10853-017-0852-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePerea-Velez YS, Gonz\u0026aacute;lez Ch\u0026aacute;vez M del CA, Carrillo-Gonz\u0026aacute;lez R, Lopez-Luna J (2022) Dissolution kinetics of citrate coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles in soil solution. Environ Sci Nano. 9:2954\u0026ndash;2965. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/D2EN00330A\u003c/span\u003e\u003cspan address=\"10.1039/D2EN00330A\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFeizi H, Rezvani Moghaddam P, Shahtahmassebi N, Fotovat A (2012) Impact of bulk and nanosized titanium dioxide (TiO\u003csub\u003e2\u003c/sub\u003e) on wheat seed germination and seedling growth. Biol Trace Elem Res 146:101\u0026ndash;106. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12011-011-9222-7\u003c/span\u003e\u003cspan address=\"10.1007/s12011-011-9222-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAntony D, Yadav R, Kalimuthu R (2021) Accumulation of phyto-mediated nano-CeO\u003csub\u003e2\u003c/sub\u003e and selenium doped CeO\u003csub\u003e2\u003c/sub\u003e on \u003cem\u003eMacrotyloma uniflorum\u003c/em\u003e (horse gram) seed by nano-priming to enhance seedling vigor. Biocatal Agric Biotechnol 31:101923. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.bcab.2021.101923\u003c/span\u003e\u003cspan address=\"10.1016/j.bcab.2021.101923\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDel Buono D, Luzi F, Tolisano C, Puglia D, Di Michele A (2022) Synthesis of a lignin/zinc oxide hybrid nanoparticles system and its application by nano-priming in maize. Nanomater 12:568. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/nano12030568\u003c/span\u003e\u003cspan address=\"10.3390/nano12030568\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVelu G, Bhattacharjee R, Rai K, Sahrawat K, Longvah T (2008) A simple and rapid screening method for grain zinc content in pearl millet. J SAT Agric Res 6:1\u0026ndash;4\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAyala Garay AV, Acosta Gallegos JA, Reyes Muro L (2021) El cultivo del frijol presente y futuro para M\u0026eacute;xico. INIFAP, Guanajuato\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHashem A, Abd_Allah EF, Alqarawi AA, Al-Huqail AA, Wirth S, Egamberdieva D (2016) The interaction between arbuscular mycorrhizal fungi and endophytic bacteria enhances plant growth of \u003cem\u003eAcacia gerrardii\u003c/em\u003e under salt stress. Front Microbiol 7:1089. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fmicb.2016.01089\u003c/span\u003e\u003cspan address=\"10.3389/fmicb.2016.01089\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSenthilkumar M, Amaresan N, Sankaranarayanan A (2021) Quantitative estimation of leghemoglobin content in legume root nodules. In: Plant-microbe interactions. Springer Protocols Handbooks. Springer, New York. pp 33\u0026ndash;35. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-1-0716-1080-0_5\u003c/span\u003e\u003cspan address=\"10.1007/978-1-0716-1080-0_5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKjeldahl J (1883) Neue methode zur bestimmung des stickstoffs in organischen k\u0026ouml;rpern. Zeitschrift f\u0026uuml;r Anal Chemie 22:366\u0026ndash;382\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKitson RE, Mellon MG (1944) Colorimetric determination of phosphorus as molybdivanadophosphoric acid. Ind Eng Chem Anal Ed 16:379\u0026ndash;383. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/i560130a017\u003c/span\u003e\u003cspan address=\"10.1021/i560130a017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMariotti F, Tom\u0026eacute; D, Mirand PP (2008) Converting nitrogen into protein\u0026mdash;beyond 6.25 and Jones\u0026rsquo; factors. Crit Rev Food Sci Nutr 48:177\u0026ndash;184. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/10408390701279749\u003c/span\u003e\u003cspan address=\"10.1080/10408390701279749\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGao Y, Shang C, Saghai Maroof MA, Biyashev RM, Grabau EA, Kwanyuen P, Burton JW, Buss GR (2007) A modified colorimetric method for phytic acid analysis in soybean. Crop Sci 47:1797\u0026ndash;1803. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2135/cropsci2007.03.0122\u003c/span\u003e\u003cspan address=\"10.2135/cropsci2007.03.0122\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDhaliwal SS, Sharma V, Shukla AK, et al (2022) Biofortification of soybean (\u003cem\u003eGlycine max\u003c/em\u003e L.) through FeSO\u003csub\u003e4\u003c/sub\u003e*7H\u003csub\u003e2\u003c/sub\u003eO to enhance yield, iron nutrition and economic outcomes in sandy loam soils of India. Agriculture 47:1797\u0026ndash;1803. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agriculture12050586\u003c/span\u003e\u003cspan address=\"10.3390/agriculture12050586\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSarwar G, Hussain N, Schmeisky H, Muhammad S (2007) Use of compost an environment friendly technology for enhancing rice-wheat production in Pakistan. Pakistan J Bot 39:1553\u0026ndash;1558\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSADER (2022) Precios de garant\u0026iacute;a a productos alimentarios b\u0026aacute;sicos, seguridad y certidumbre a productores. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.gob.mx/agricultura/articulos/precios-de-garantia-a-productos-alimentarios-basicos-seguridad-y-certidumbre-a-productores\u003c/span\u003e\u003cspan address=\"https://www.gob.mx/agricultura/articulos/precios-de-garantia-a-productos-alimentarios-basicos-seguridad-y-certidumbre-a-productores\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Accessed 9 October 2022\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePerea-V\u0026eacute;lez YS, Carrillo-Gonz\u0026aacute;lez R, Gonz\u0026aacute;lez-Ch\u0026aacute;vez M del CA, Vangronsveld J, Monasterio IO, Tapia Maruri D (2023) Citrate-coated cobalt ferrite nanoparticles for the nano-enabled biofortification of wheat. Food Funct. 14:4017\u0026ndash;4035. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1039/D2FO03835H\u003c/span\u003e\u003cspan address=\"10.1039/D2FO03835H\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eInternational Monetary Fund (2021) Representative exchange rates for selected currencies for september 2021. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.imf.org/external/np/fin/data/rms_mth.aspx?SelectDate=2021-09-30\u0026amp;reportType=REP\u003c/span\u003e\u003cspan address=\"https://www.imf.org/external/np/fin/data/rms_mth.aspx?SelectDate=2021-09-30\u0026amp;reportType=REP\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Accessed 9 October 2022\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR Core Team (2020) R: A language and environment for statistical computing. Vienna, Austria, 2020.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRomano A, Stevanato P (2020) Germination data analysis by time-to-event approaches. Plants. 9:617. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/plants9050617\u003c/span\u003e\u003cspan address=\"10.3390/plants9050617\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOnofri A, Benincasa P, Mesgaran MB, Ritz C (2018) Hydrothermal-time-to-event models for seed germination. Eur J Agron 101:129\u0026ndash;139. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.eja.2018.08.011\u003c/span\u003e\u003cspan address=\"10.1016/j.eja.2018.08.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOnofri A, Mesgaran MB, Ritz C (2022) A unified framework for the analysis of germination, emergence, and other time-to-event data in weed science. Weed Sci 70:259\u0026ndash;271. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/wsc.2022.8\u003c/span\u003e\u003cspan address=\"10.1017/wsc.2022.8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhan MN, Fu C, Li J, Tao Y, Li Y, Hu J, Chen L, Khan Z, Wu H, Li Z (2023) Seed nanopriming: How do nanomaterials improve seed tolerance to salinity and drought? Chemosphere 310:13691. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.chemosphere.2022.136911\u003c/span\u003e\u003cspan address=\"10.1016/j.chemosphere.2022.136911\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAcharya A, Pal PK (2020) Agriculture nanotechnology: Translating research outcome to field applications by influencing environmental sustainability. NanoImpact 19:100232. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.impact.2020.100232\u003c/span\u003e\u003cspan address=\"10.1016/j.impact.2020.100232\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSavassa SM, Castillo-Michel H, Pradas del Real AE, Reyes-Herrera J, Marques JPR, de Carvalho HWP (2021) Ag nanoparticles enhancing \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e seedling development: understanding nanoparticle migration and chemical transformation across the seed coat. Environ Sci Nano 8:493\u0026ndash;50. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1039/D0EN00959H\u003c/span\u003e\u003cspan address=\"10.1039/D0EN00959H\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSavassa SM, Duran NM, Rodrigues ES, de Almeida E, van Gestel CAM, Bompadre TF V, P. de Carvalho HW (2018) Effects of ZnO nanoparticles on \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e germination and seedling development determined by X-ray spectroscopy. ACS Appl Nano Mater 1:6414\u0026ndash;6426. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/acsanm.8b01619\u003c/span\u003e\u003cspan address=\"10.1021/acsanm.8b01619\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eForest V, Cottier M, Pourchez J (2015) Electrostatic interactions favor the binding of positive nanoparticles on cells: A reductive theory. Nano Today 10:677\u0026ndash;680. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.nantod.2015.07.002\u003c/span\u003e\u003cspan address=\"10.1016/j.nantod.2015.07.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAfzal S, Sharma D, Singh NK (2021) Eco-friendly synthesis of phytochemical-capped iron oxide nanoparticles as nano-priming agent for boosting seed germination in rice (\u003cem\u003eOryza sativa\u003c/em\u003e L.). Environ Sci Pollut Res 28:40275\u0026ndash;40287. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11356-020-12056-5\u003c/span\u003e\u003cspan address=\"10.1007/s11356-020-12056-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIghaiee Oskoiee A, Ghanbari AA, Mirhadi MJ (2021) Effects of iron nanoparticles on seed germination of bean genotypes. SSRN Electron J. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2139/ssrn.3899301\u003c/span\u003e\u003cspan address=\"10.2139/ssrn.3899301\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKandhol N, Singh VP, Ramawat N, Prasad R, Chauhan DK, Sharma S, Grillo R, Sahi S, Peralta-Videa J, Tripathi DK (2022) Nano-priming: Impression on the beginner of plant life. Plant Stress 5:100091. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.stress.2022.100091\u003c/span\u003e\u003cspan address=\"10.1016/j.stress.2022.100091\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePawar VA, Ambekar JD, Kale BB, Apte SK, Laware SL (2019) Response in chickpea (\u003cem\u003eCicer arietinum\u003c/em\u003e L.) seedling growth to seed priming with iron oxide nanoparticles. Int J Biosci 14:82\u0026ndash;91. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.12692/ijb/14.3.82-91\u003c/span\u003e\u003cspan address=\"10.12692/ijb/14.3.82-91\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKabata-Pendias A (2010) Trace Elements in Soils and Plants (4th edition). CRC Press, Boca Raton. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1201/b10158\u003c/span\u003e\u003cspan address=\"10.1201/b10158\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePlank CO, Kissel DE (1989) Plant Analysis Handbook for Georgia\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRosenblueth M, Mart\u0026iacute;nez-Romero E (2004) \u003cem\u003eRhizobium e\u003c/em\u003etli maize populations and their competitiveness for root colonization. Arch Microbiol 181:337\u0026ndash;344. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00203-004-0661-9\u003c/span\u003e\u003cspan address=\"10.1007/s00203-004-0661-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang M-X, Zhao L-Y, He Y-Y, Hu J-P, Hu G-W, Zhu Y, Khan A, Xiong Y-C, Zhang J-L (2024) Potential roles of iron nanomaterials in enhancing growth and nitrogen fixation and modulating rhizomicrobiome in alfalfa (\u003cem\u003eMedicago sativa\u003c/em\u003e L.). Bioresour Technol 391:129987. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.biortech.2023.129987\u003c/span\u003e\u003cspan address=\"10.1016/j.biortech.2023.129987\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFedorova EE, Coba de la Pe\u0026ntilde;a T, Lara-Dampier V, Trifonova NA, Kulikova O, Pueyo JJ, Lucas MM (2021) Potassium content diminishes in infected cells of \u003cem\u003eMedicago truncatula\u003c/em\u003e nodules due to the mislocation of channels MtAKT1 and MtSKOR/GORK. J Exp Bot 72:1336\u0026ndash;1348. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/jxb/eraa508\u003c/span\u003e\u003cspan address=\"10.1093/jxb/eraa508\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang J, Cao X, Wang C, Chen F, Feng Y, Yue L, Wang Z, Xing B (2022) Fe-based nanomaterial-induced root nodulation is modulated by flavonoids to improve soybean (\u003cem\u003eGlycine max\u003c/em\u003e) growth and quality. ACS Nano 16:21047\u0026ndash;21062. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/acsnano.2c08753\u003c/span\u003e\u003cspan address=\"10.1021/acsnano.2c08753\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMontes-Luz B, Conrado AC, Ellingsen JK, Monteiro RA, de Souza EM, Stacey G (2023) Acetylene reduction assay: A measure of nitrogenase activity in plants and bacteria. Curr Protoc. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/cpz1.766\u003c/span\u003e\u003cspan address=\"10.1002/cpz1.766\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi M, Gao L, White JC, Haynes CL, O\u0026rsquo;Keefe TL, Rui Y, Ullah S, Guo Z, Lynch I, Zhang P (2023) Nano-enabled strategies to enhance biological nitrogen fixation. Nat Nanotechnol 18:688\u0026ndash;69. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41565-023-01392-5\u003c/span\u003e\u003cspan address=\"10.1038/s41565-023-01392-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCh\u0026aacute;vez-Mendoza C, S\u0026aacute;nchez E (2017) Bioactive compounds from mexican varieties of the common bean (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e): Implications for health. Molecules 22:1360. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules22081360\u003c/span\u003e\u003cspan address=\"10.3390/molecules22081360\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHuertas R, Karpinska B, Ngala S, et al (2022) Biofortification of common bean (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L.) with iron and zinc: Achievements and challenges. Food Energy Secur 12:e406. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/fes3.406\u003c/span\u003e\u003cspan address=\"10.1002/fes3.406\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDiaz S, Polania J, Ariza-Suarez D, Cajiao C, Grajales M, Raatz B, Beebe SE (2022) Genetic correlation between Fe and Zn biofortification and yield components in a common bean (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L.). Front Plant Sci 12:739033. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2021.739033\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2021.739033\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBeebe S (2020) Biofortification of common bean for higher iron concentration. Front Sustain Food Syst 4:573449. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fsufs.2020.573449\u003c/span\u003e\u003cspan address=\"10.3389/fsufs.2020.573449\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEspinoza-Garc\u0026iacute;a N, Mart\u0026iacute;nez-Mart\u0026iacute;nez R, Ch\u0026aacute;vez-Servia JL, Vera-Guzm\u0026aacute;n AM, Carrillo-Rodr\u0026iacute;guez JC, Heredia-Garc\u0026iacute;a E, Velasco-Velasco VA (2016) Contenido de minerales en semilla de poblaciones nativas de frijol com\u0026uacute;n (Phaseolus vulgaris L.). Rev Fitotec Mex 39:215\u0026ndash;223\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHarvestPlus (2022) Hight iron beans. A food that can change your business and the world naturally. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.harvestplus.org/wp-content/uploads/2022/01/Iron-Beans.pdf\u003c/span\u003e\u003cspan address=\"https://www.harvestplus.org/wp-content/uploads/2022/01/Iron-Beans.pdf\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Accessed 9 October 2022\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHummel M, Talsma EF, Taleon V, Londo\u0026ntilde;o L, Brychkova G, Gallego S, Raatz B, Spillane C (2020) Iron, zinc and phytic acid retention of biofortified, low phytic acid, and conventional bean varieties when preparing common household recipes. Nutrients 12:658. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/nu12030658\u003c/span\u003e\u003cspan address=\"10.3390/nu12030658\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCastro-Alba V, Lazarte CE, Bergenst\u0026aring;hl B, Granfeldt Y (2019) Phytate, iron, zinc, and calcium content of common Bolivian foods and their estimated mineral bioavailability. Food Sci Nutr 7:2854\u0026ndash;2865. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/fsn3.1127\u003c/span\u003e\u003cspan address=\"10.1002/fsn3.1127\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSiimes N, Sharp EL, Lewis N, Kah M (2022) Determining acceptance and rejection of nano-enabled agriculture: A case study of the New Zealand wine industry. NanoImpact 28:100432. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.impact.2022.100432\u003c/span\u003e\u003cspan address=\"10.1016/j.impact.2022.100432\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarg M, Sharma N, Sharma S, Kapoor P, Kumar A, Chunduri V, Arora P (2018) Biofortified crops generated by breeding, agronomy, and transgenic approaches are improving lives of millions of people around the world. Front Nutr 5:12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fnut.2018.00012\u003c/span\u003e\u003cspan address=\"10.3389/fnut.2018.00012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAcharya P, Jayaprakasha GK, Crosby KM, Jifon JL, Patil BS (2020) Nanoparticle-mediated seed priming improves germination, growth, yield, and quality of watermelons (\u003cem\u003eCitrullus lanatus\u003c/em\u003e) at multi-locations in Texas. Sci Rep 10:5037. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-020-61696-7\u003c/span\u003e\u003cspan address=\"10.1038/s41598-020-61696-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePaul S, Dey S, Kundu R (2022) Seed priming: an emerging tool towards sustainable agriculture. Plant Growth Regul 97:215\u0026ndash;234. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10725-021-00761-1\u003c/span\u003e\u003cspan address=\"10.1007/s10725-021-00761-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\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":"journal-of-nanoparticle-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"nano","sideBox":"Learn more about [Journal of Nanoparticle Research](http://link.springer.com/journal/11051)","snPcode":"11051","submissionUrl":"https://submission.nature.com/new-submission/11051/3","title":"Journal of Nanoparticle Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"seed priming, nano-enabled agriculture, nanotechnology cost, biofortification","lastPublishedDoi":"10.21203/rs.3.rs-4578599/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4578599/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNano-priming is an emerging application of nanotechnology in agriculture intending to increase crop yield and nutritional quality, while reducing fertilizer applications. This study aimed to investigate the effects of seed priming with citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles (NPs) suspensions (10, 20, and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) on the life cycle of the \u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L. OTI cultivar and evaluate the technology costs. The effect of nano-priming was assessed in the germination, flowering, and harvest stages. Unprimed and hydro-primed seeds were negative and positive controls, respectively. Nano-priming with CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs had no effect neither on the germination nor on plant nutrition (in the flowering stage) of OTI beans compared to unprimed and hydro-primed seeds. In contrast, nitrogenase activity (343.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1 \u0026micro;mol h\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of C\u003csub\u003e2\u003c/sub\u003eH\u003csub\u003e4\u003c/sub\u003e ) was detected in the plants from the 40 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e nano-primed seeds. The K concentration of progeny seeds from nano-priming with 10, 20, and 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e increased significantly by 3%, 16%, and 13% compared to the control seeds. The Zn concentration in the seeds from nano-priming with 10 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e was 27% higher than in the control and 28% higher than in the hydro-primed seeds. When nano-priming with 40 mg NPs L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, the Zn concentration was 5% and 6% higher than the control and hydro-primed seeds. The calculated cost of nano-priming seeds per ha ranged from 121 to 143 USD. In this regard, nano-priming of bean seeds with citrate-coated CoFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs could be a low-cost approach to achieve nutritional security and agricultural sustainability.\u003c/p\u003e","manuscriptTitle":"Nano-priming of Phaseolus vulgaris OTI cultivar with cobalt ferrite nanoparticles enhances the mineral composition of progeny seeds","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-04 17:48:49","doi":"10.21203/rs.3.rs-4578599/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-07-10T03:26:01+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-09T17:32:32+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"10083895796663553489786429583921874479","date":"2024-06-26T10:18:24+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-06-21T02:52:35+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-06-19T20:58:11+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-06-18T14:53:01+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Nanoparticle Research","date":"2024-06-14T00:31:03+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-nanoparticle-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"nano","sideBox":"Learn more about [Journal of Nanoparticle Research](http://link.springer.com/journal/11051)","snPcode":"11051","submissionUrl":"https://submission.nature.com/new-submission/11051/3","title":"Journal of Nanoparticle Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"e951996a-9950-41df-ab6c-9c1ac6f3acd6","owner":[],"postedDate":"July 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-09-02T16:07:04+00:00","versionOfRecord":{"articleIdentity":"rs-4578599","link":"https://doi.org/10.1007/s11051-024-06101-4","journal":{"identity":"journal-of-nanoparticle-research","isVorOnly":false,"title":"Journal of Nanoparticle Research"},"publishedOn":"2024-08-30 15:57:26","publishedOnDateReadable":"August 30th, 2024"},"versionCreatedAt":"2024-07-04 17:48:49","video":"","vorDoi":"10.1007/s11051-024-06101-4","vorDoiUrl":"https://doi.org/10.1007/s11051-024-06101-4","workflowStages":[]},"version":"v1","identity":"rs-4578599","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4578599","identity":"rs-4578599","version":["v1"]},"buildId":"zQwnuV7TCBrMSSSToR1PI","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}