Integrating transformer-based machine learning with SERS technology for the analysis of hazardous pesticides in spinach.

This study introduces an innovative strategy for the rapid and accurate identification of pesticide residues in agricultural products by combining surface-enhanced Raman spectroscopy (SERS) with a state-of-the-art transformer model, termed SERSFormer. Gold-silver core-shell nanoparticles were synthesized and served as high-performance SERS substrates, which possess well-defined structures, uniform dispersion, and a core-shell composition with an average diameter of 21.44 ± 4.02 nm, as characterized by TEM-EDS. SERSFormer employs sophisticated, task-specific data processing techniques and CNN embedders, powered by an architecture features weight-shared multi-head self-attention transformer encoder layers. The SERSFormer model demonstrated exceptional proficiency in qualitative analysis, successfully classifying six categories, including five pesticides (coumaphos, oxamyl, carbophenothion, thiabendazole, and phosmet) and a control group of spinach data, with 98.4% accuracy. For quantitative analysis, the model accurately predicted pesticide concentrations with a mean absolute error of 0.966, a mean squared error of 1.826, and an R2 score of 0.849. This novel approach, which combines SERS with machine learning and is supported by robust transformer models, showcases the potential for real-time pesticide detection to improve food safety in the agricultural and food industries.

It takes two to attach - endo-1,3-ß-d-glucanase as a potential receptor of mannose-independent, FimH-dependent Salmonella Typhimurium binding to spinach leaves.

Currently, fresh, unprocessed food has become a relevant element of the chain of transmission of enteropathogenic infections. To survive on a plant surface and further spread the infections, pathogens like Salmonella have to attach stably to the leaf surface. Adhesion, driven by various virulence factors, including the most abundant fim operon encoding type 1 fimbriae, is usually an initial step of infection, preventing physical removal of the pathogen. Adhesion properties of Salmonella's type 1 fimbriae and its FimH adhesin were investigated intensively in the past. However, there is a lack of knowledge regarding its role in interaction with plant cells. Understanding the mechanisms and structures involved in such interaction may facilitate efforts to decrease the risk of contamination and increase fresh food safety. Here, we applied Salmonella genome site-directed mutagenesis, adhesion assays, protein-protein interactions, and biophysics methods based on surface plasmon resonance to unravel the role of FimH adhesin in interaction with spinach leaves. We show that FimH is at least partially responsible for Salmonella binding to spinach leaves, and this interaction occurs in a mannose-independent manner. Importantly, we identified a potential FimH receptor as endo-1,3-ß-d-Glucanase and found that this interaction is strong and specific, with a dissociation constant in the nanomolar range. This research advances our comprehension of Salmonella's interactions with plant surfaces, offering insights that can aid in minimizing contamination risks and improving the safety of fresh, unprocessed foods.

Inoculation of heavy metal resistant bacteria alleviated heavy metal-induced oxidative stress biomarkers in spinach (Spinacia oleracea L.).

Most vegetable crops are severely affected by the uptake of heavy metals from the soil. Heavy metals in vegetable bodies generate reactive oxygen species (ROS) that unbalance the antioxidant defense system. This study was initiated to determine the physiological and biochemical characteristics of spinach plants grown on soil contaminated with heavy metals and responding to Bacillus cereus and Bacillus aerius were isolated from soil contaminated with heavy metals. Heavy metal contamination led to a significant reduction in seed germination, seedling biomass, protein, and total nitrogen content of spinach plants grown in contaminated soils compared to control soils. In contrast, a significant increase in the content of metallothioneins and antioxidant enzymes was observed. Plants inoculated with B. cereus and B. aerius significantly reduced the oxidative stress induced by heavy metals by improving seed germination (%), seedling growth, nitrogen, and protein content. The content of metallothioneins and the activities of antioxidant enzymes were reduced in spinach plants grown from seeds inoculated with bacterial strains. In addition, plants inoculated with, B. cereus and B. aerius showed greater stomata opening than plants grown on soil contaminated with heavy metals, whose stomata were almost closed. These results suggested that both bacterial strains enhanced plant growth by reducing oxidative stress caused by metals.

Effective degradation of synthetic micropollutants and real textile wastewater via a visible light-activated persulfate system using novel spinach leaf-derived biochar.

A novel biochar (BC), derived from spinach leaves, was utilized as an activator for persulfate (PS) in the degradation of methylene blue (MB) dye under visible light conditions. Thorough analyses were conducted to characterize the physical and chemical properties of the biochar. The (BC + light)/PS system exhibited superior MB degradation efficiency at 83.36%, surpassing the performance of (BC + light)/hydrogen peroxide and (BC + light)/peroxymonosulfate systems. The optimal conditions were ascertained through the implementation of response surface methodology. Moreover, the (BC + light)/PS system demonstrated notable degradation ratios of 90.82%, 81.88%, and 84.82% for bromothymol blue dye, paracetamol, and chlorpyrifos, respectively, under optimal conditions. The predominant reactive species responsible for MB degradation were identified as sulfate radicals. Notably, the proposed system consistently achieved high removal efficiencies of 99.02%, 96.97%, 94.94%, 92%, and 90.35% for MB in five consecutive runs. The applicability of the suggested system was further validated through its effectiveness in treating real textile wastewater, exhibiting a substantial MB removal efficiency of 98.31% and dissolved organic carbon mineralization of 87.49%.

A Genome-Wide Association Study Reveals the Genetic Mechanisms of Nutrient Accumulation in Spinach.

Spinach is a significant source of vitamins, minerals, and antioxidants. These nutrients make it delicious and beneficial for human health. However, the genetic mechanism underlying the accumulation of nutrients in spinach remains unclear. In this study, we analyzed the content of chlorophyll a, chlorophyll b, oxalate, nitrate, crude fiber, soluble sugars, manganese, copper, and iron in 62 different spinach accessions. Additionally, 3,356,182 high-quality, single-nucleotide polymorphisms were found using resequencing and used in a genome-wide association study. A total of 2077 loci were discovered that significantly correlated with the concentrations of the nutritional elements. Data mining identified key genes in these intervals for four traits: chlorophyll, oxalate, soluble sugar, and Fe. Our study provides insights into the genetic architecture of nutrient variation and facilitates spinach breeding for good nutrition.

QTL analysis of femaleness in monoecious spinach and fine mapping of a major QTL using an updated version of chromosome-scale pseudomolecules.

Although spinach is predominantly dioecious, monoecious plants with varying proportions of female and male flowers are also present. Recently, monoecious inbred lines with highly female and male conditions have been preferentially used as parents for F1-hybrids, rather than dioecious lines. Accordingly, identifying the loci for monoecism is an important issue for spinach breeding. We here used long-read sequencing and Hi-C technology to construct SOL_r2.0_pseudomolecule, a set of six pseudomolecules of spinach chromosomes (total length: 879.2 Mb; BUSCO complete 97.0%) that are longer and more genetically complete than our previous version of pseudomolecules (688.0 Mb; 81.5%). Three QTLs, qFem2.1, qFem3.1, and qFem6.1, responsible for monoecism were mapped to SOL_r2.0_pseudomolecule. qFem3.1 had the highest LOD score and corresponded to the M locus, which was previously identified as a determinant of monoecious expression, by genetic analysis of progeny from female and monoecious plants. The other QTLs were shown to modulate the ratio of female to male flowers in monoecious plants harboring a dominant allele of the M gene. Our findings will enable breeders to efficiently produce highly female- and male-monoecious parental lines for F1-hybrids by pyramiding the three QTLs. Through fine-mapping, we narrowed the candidate region for the M locus to a 19.5 kb interval containing three protein-coding genes and one long non-coding RNA gene. Among them, only RADIALIS-like-2a showed a higher expression in the reproductive organs, suggesting that it might play a role in reproductive organogenesis. However, there is no evidence that it is involved in the regulation of stamen and pistil initiation, which are directly related to the floral sex differentiation system in spinach. Given that auxin is involved in reproductive organ formation in many plant species, genes related to auxin transport/response, in addition to floral organ formation, were identified as candidates for regulators of floral sex-differentiation from qFem2.1 and qFem6.1.

Identification and reactivity of pigments in prominent vegetable leaves of Basella alba L. var. 'Rubra' (Malabar spinach).

The unique profiles of betacyanins as well as their stability and antioxidant activity in purple leaf extracts of the fast-growing, soft-stemmed vine Basella alba L. var. 'Rubra', known as Malabar spinach, are partly characterized for the first time. The distribution of gomphrenin and its acylated derivatives in the leaves is completely different from the profiles of the pigments in the fruits. The most abundant acylated pigment in leaves (24%) turned out 6'-O-E-sinapoyl-gomphrenin (gandolin), however, the most significant difference in the pigment profiles is a presence of two novel pigments tentatively identified as highly abundant 6'-O-(3,4-dimethoxy-E-cinnamoyl)-gomphrenin and 6'-O-(3,4,5-trimethoxy-E-cinnamoyl)-gomphrenin as well as their isoforms. Significant degradation of the pigments in the fruit extracts under the impact of selected metal cations and UV-Vis irradiation as well as high protective activity of the leaf extract matrix were observed. Partial chromatographic purification of the leaf extract resulted in an increase of the pigment concentration which was correlated positively with the increased antioxidant activity of obtained fractions.

Basella alba L. (Malabar Spinach) as an Abundant Source of Betacyanins: Identification, Stability, and Bioactivity Studies on Natural and Processed Fruit Pigments.

The antioxidant and anti-inflammatory activities of acylated and decarboxylated gomphrenins, as well as Basella alba L. fruit extract, were investigated in relation to gomphrenin, known for its high biological potential. The most abundant natural acylated gomphrenins, namely, 6'-O-E-caffeoyl-gomphrenin (malabarin) and 6'-O-E-4-coumaroyl-gomphrenin (globosin), were isolated from B. alba extract for the studies. In addition, controlled thermal decarboxylation of gomphrenin in the purified B. alba extract at 65-75 °C resulted in the formation of the most prevalent decarboxylated products, including 17-decarboxy-gomphrenin and 2,17-bidecarboxy-gomphrenin, along with their isoforms. The structures of the decarboxylated pigments were confirmed by NMR analyses. Exploring the matrix effect on pigment reactivity revealed a tremendous increase in the stability of all betacyanins after the initial stage of extract purification using a cation exchanger under various conditions. This indicates the removal of a substantial portion of the unfavorable matrix from the extract, which presumably contains reactive species that could otherwise degrade the pigments. Furthermore, the high concentration of citrates played a significant role in favoring the formation of 2-decarboxy-gomphrenin to a considerable extent. In vitro screening experiments revealed that the tested compounds demonstrated strong anti-inflammatory properties in lipopolysaccharide (LPS)-activated human macrophages. This effect encompassed the selective inhibition of cytokine and chemokine release from activated macrophages, modulation of the chemotactic activity of immune cells, and the regulation of tissue remodeling mediators' release.

Temporal phenotypic variation of spinach root traits and its relation to shoot performance.

The root system is important for the growth and development of spinach. To reveal the temporal variability of the spinach root system, root traits of 40 spinach accessions were measured at three imaging times (20, 30, and 43 days after transplanting) in this study using a non-destructive and non-invasive root analysis system. Results showed that five root traits were reliably measured by this system (RootViz FS), and two of which were highly correlated with manually measured traits. Root traits had higher variations than shoot traits among spinach accessions, and the trait of mean growth rate of total root length had the largest coefficients of variation across the three imaging times. During the early stage, only tap root length was weakly correlated with shoot traits (plant height, leaf width, and object area (equivalent to plant surface area)), whereas in the third imaging, root fresh weight, total root length, and root area were strongly correlated with shoot biomass-related traits. Five root traits (total root length, tap root length, total root area, root tissue density, and maximal root width) showed high variations with coefficients of variation values (CV ≥ 0.3, except maximal root width) and high heritability (H2 > 0.6) among the three stages. The 40 spinach accessions were classified into five subgroups with different growth dynamics of the primary and lateral roots by cluster analysis. Our results demonstrated the potential of in-situ phenotyping to assess dynamic root growth in spinach and provide new perspectives for biomass breeding based on root system ideotypes.

Circadian-based approach for improving physiological, phytochemical and chloroplast proteome in Spinacia oleracea under salinity stress and light emitting diodes.

Salt stress is a recognized annihilating abiotic stress that has a significant impact on agricultural and horticulture crop productivity. Plant development faces three distinct dangers as a result of salt stress: oxidative stress, osmotic stress, and ionic toxicity. It has been shown that plants can forecast diurnal patterns using the circadian clock; moreover, they can manage their defensive mechanism for the detoxification of reactive oxygen species (ROS). Circadian rhythmicity in gene expression assembles transcription and translation feedback networks to govern plant shape, physiology, cellular and molecular activities. Both external and internal variables influence the systemic rhythm via input routes. The Malav Jyoti (MJ) and Delhi Green (DG) genotypes of spinach (Spinacia oleracea) were grown in the plant growth chamber. The chamber had an optimized temperature of 25 °C and humidity of 65% containing light emitting diode (LED) having Red: Blue: white (one side) and White fluorescent (other side) under salinity stress. The samples were collected on the basis of 4 h intervals of circadian hours (0 h, 4 h, 8 h and 12 h) during Day-10 and Day-20 of salt treatments. Under salt stress, the circadian and light-emitting diode-based strategy had a substantial influence on spinach's anti-oxidative responses, stomatal movement, CO2 assimilation, PS-I and II efficiency, phytochrome pigment efficiency, and photosynthesis. Based on the findings of the free radical scavenging enzyme tests, the photoperiodic hours for the proteome analysis were set to 11 am and 3 pm on Day-20. When compared to white fluorescent, this study found that LED has the capacity to influence the entrainment cues of the circadian clock in the cultivation of salt-sensitive spinach genotypes. According to our findings, changing the cellular scavenging mechanism and chloroplast proteome has increased the survival rate of spinach genotypes under LED when compared to white fluorescent.

Metabolomics reveals how spinach plants reprogram metabolites to cope with intense stress responses induced by photoaged polystyrene nanoplastics (PSNPs).

While land-based sources have been recognized as significant long-term sinks for micro- and nanoplastics, there is limited knowledge about the uptake, translocation, and phytotoxicity of nanoplastics (NPs) in terrestrial environments, especially aged NPs. In this study, we investigated the impact of aged polystyrene nanoplastics (PSNPs) on the uptake, physiology, and metabolism of spinach. Our findings revealed that both pristine and aged PSNPs can accumulate in the roots and subsequently translocate to the aboveground tissues, thereby influencing numerous key growth indicators in spinach plants. A more pronounced impact was observed in the treatment of aged PSNPs, triggering more significant and extensive changes in metabolite levels. Furthermore, alterations in targeted pathways, specifically aminoacyl-tRNA biosynthesis and phenylpropanoid biosynthesis, were induced by aged PSNPs, while pristine PSNPs influenced pathways related to sulfur metabolism, biosynthesis of unsaturated fatty acids, and tryptophan metabolism. Additionally, tissue-specific responses were observed at the metabolomics level in both roots and leaves. These results highlight the existence of diverse and tissue-specific metabolic responses in spinach plants exposed to pristine and aged PSNPs, providing insights into the mechanisms of defense and detoxification against NP-induced stress.

Fates of attached E. coli o157:h7 on intact leaf surfaces revealed leafy green susceptibility.

Leafy greens, especially lettuce, are repeatedly linked to foodborne outbreaks. This paper studied the susceptibility of different leafy greens to human pathogens. Five commonly consumed leafy greens, including romaine lettuce, green-leaf lettuce, baby spinach, kale, and collard, were selected by their outbreak frequencies. The behavior of E. coli O157:H7 87-23 on intact leaf surfaces and in their lysates was investigated. Bacterial attachment was positively correlated with leaf surface roughness and affected by the epicuticular wax composition. At room temperature, E. coli O157:H7 had the best growth potentials on romaine and green-leaf lettuce surfaces. The bacterial growth was positively correlated with stomata size and affected by epicuticular wax compositions. At 37 °C, E. coli O157:H7 87-23 was largely inhibited by spinach and collard lysates, and it became undetectable in kale lysate after 24 h of incubation. Kale and collard lysates also delayed or partially inhibited the bacterial growth in TSB and lettuce lysate at 37 °C, and they sharply reduced the E. coli O157:H7 population on green leaf lettuce at 4 °C. In summary, the susceptibility of leafy greens to E. coli O157:H7 is determined by a produce-specific combination of physiochemical properties and temperature.

Genome-Wide Identification and Characterization of MYB Gene Family and Analysis of Its Sex-Biased Expression Pattern in Spinacia oleracea L.

The members of the myeloblastosis (MYB) family of transcription factors (TFs) participate in a variety of biological regulatory processes in plants, such as circadian rhythm, metabolism, and flower development. However, the characterization of MYB genes across the genomes of spinach Spinacia oleracea L. has not been reported. Here, we identified 140 MYB genes in spinach and described their characteristics using bioinformatics approaches. Among the MYB genes, 54 were 1R-MYB, 80 were 2R-MYB, 5 were 3R-MYB, and 1 was 4R-MYB. Almost all MYB genes were located in the 0-30 Mb region of autosomes; however, the 20 MYB genes were enriched at both ends of the sex chromosome (chromosome 4). Based on phylogeny, conserved motifs, and the structure of genes, 2R-MYB exhibited higher conservation relative to 1R-MYB genes. Tandem duplication and collinearity of spinach MYB genes drive their evolution, enabling the functional diversification of spinach genes. Subcellular localization prediction indicated that spinach MYB genes were mainly located in the nucleus. Cis-acting element analysis confirmed that MYB genes were involved in various processes of spinach growth and development, such as circadian rhythm, cell differentiation, and reproduction through hormone synthesis. Furthermore, through the transcriptome data analysis of male and female flower organs at five different periods, ten candidate genes showed biased expression in spinach males, suggesting that these genes might be related to the development of spinach anthers. Collectively, this study provides useful information for further investigating the function of MYB TFs and novel insights into the regulation of sex determination in spinach.

Nickel uptake in leafy greens from contaminated soil: an investigation into phytoavailability and health risk assessment using in vitro digestion model.

Nickel (Ni) is a toxic metal that not only pollutes the environment but also causes harmful impacts on plant growth and human health. Therefore, it is crucial to assess the relationship between the phytoavailability of Ni in soil and its accumulation in edible and non-edible parts of vegetables. A pot experiment was conducted to investigate Ni uptake in three different leafy vegetables, spinach (Spinacia oleracea L.), lettuce (Lactuca sativa L.), and fenugreek (Trigonella foenum-graecum L.), grown in soil artificially contaminated with Ni at three different treatment levels (100 mg kg-1, 200 mg kg-1, and 300 mg kg-1). The potential dietary toxicity of these vegetables in humans was examined by using an in vitro digestion model. The lowest and highest chlorophyll contents were detected in lettuce at 300 mg kg-1 of Ni concentration and in control plants of spinach. Their values were 34.16 ± 3.01 (SPAD unit) and 53 ± 3.7673 (SPAD unit), respectively. Among the three vegetables, lettuce and spinach at 300 mg kg-1 exhibited the highest accumulation of Ni, with 43 mg kg-1 in edible parts and 182 mg kg-1 in non-edible parts. Furthermore, health risk index (HRI) values were found to be > 1 for lettuce and fenugreek at Ni concentrations of 200 and 300 mg kg-1 for both children and adults. The average bioaccessibility of Ni in lettuce, fenugreek, and spinach during the gastrointestinal phase was 32-23%, 24-10%, and 45-37%, respectively, at a Ni concentration of 300 mg kg-1. All three vegetables grown on Ni-contaminated soil may potentially contribute to food chain toxicity. The HRI values being > 1 suggest that these vegetables are unsafe for consumption. Monitoring of Ni concentration in leafy vegetables is essential to minimize human health risks associated with food chain contamination.

Capillary electrophoresis tandem mass spectrometry to determine multiclass cyanotoxins in reservoir water and spinach samples.

Cyanotoxins constitute a group of toxic secondary metabolites, the presence of which in any water body poses a major health risk. Moreover, advanced organisms such as edible plants exposed to these toxins, are a possible pathway for human exposure. Green analytical chemistry is demanding environmentally friendly analytical techniques. In this sense, we propose the use of capillary electrophoresis coupled to tandem mass spectrometry (CE-MS/MS) to determine a mixture of eight cyanotoxins belonging to three different classes: cyclic peptides (microcystin-LR, microcystin-RR and nodularin), alkaloids (cylindrospermopsin and anatoxin-a) and three isomeric non-protein amino acids (ß-methylamino-l-alanine, 2,4-diaminobutyric acid and N-(2-aminoethyl)glycine). Separation was achieved by using an acidic background electrolyte consisting of 2 M formic acid and 20% acetonitrile in water. Parameters affecting MS/MS detection and the sheath-liquid interface were also studied. Finally, a combination of pH-junction, field-amplified sample stacking (FASS) and acid barrage as online preconcentration strategies, was employed to improve sensitivity and efficiency. The online preconcentration applied, in combination with a dual cartridge solid-phase extraction (SPE) system, allows to obtain limits of detection in the very low range of µg·L-1 for these multiclass cyanotoxins in reservoir water samples (from 0.005 to 0.10 µg·L-1). Furthermore, for the first time cyanotoxins are analysed in spinach samples through CE-MS/MS using the same SPE procedure, following lyophilisation and solid-liquid extraction with 6 mL 80 % aqueous MeOH.

Physiological and biochemical effects of biochar nanoparticles on spinach exposed to salinity and drought stresses.

The utilization of nanobiochar in agricultural practices has garnered substantial interest owing to its promising potential. Its nano-size particles possess an enhanced ability to infiltrate plant cells, potentially instigating biochemical and physiological responses that augment stress tolerance. In our study, we aimed to assess the impact and extent of exogenously applied nanobiochar on the growth dynamics and antioxidative responses in Spinacia oleracea L. (spinach) plants subjected to salt stress (50 mM NaCl) and drought stress (maintained at 60% field capacity) compared with respective controls (0 mM NaCl and 100% field capacity). Following a 15-day exposure to stress conditions, nanobiochar solution (at concentrations of 0, 1, 3, and 5% w/v) was sprayed on spinach plants at weekly intervals (at 14, 21, and 28 days after sowing). The foliar application of nanobiochar markedly improved biomass, net assimilation rate, leaf area, and various other growth parameters under drought and salinity stress conditions. Notably, the application of 3% nanobiochar caused the most significant enhancement in growth traits, photosynthetic pigments, and nutrient content, indicating its efficiency in directly supplying nutrients to the foliage. Furthermore, under drought stress conditions, the application of 3% nanobiochar elicited a notable 62% increase in catalase activity, a two-fold rise in peroxidase activity, and a 128% increase in superoxide dismutase activity compared to the control (without nanobiochar). Additionally, nanobiochar application enhanced membrane stability, evidenced by reduced lipid peroxidation and electrolyte leakage. The foliar application of 3% nanobiochar was found as a promising strategy to significantly enhance spinach growth parameters, nutrient assimilation, and antioxidative defense mechanisms, particularly under conditions of drought and salinity stress.

Oversampling methods for machine learning model data training to improve model capabilities to predict the presence of Escherichia coli MG1655 in spinach wash water.

We assessed the efficacy of oversampling techniques to enhance machine learning model performance in predicting Escherichia coli MG1655 presence in spinach wash water. Three oversampling methods were applied to balance two datasets, forming the basis for training random forest (RF), support vector machines (SVMs), and binomial logistic regression (BLR) models. Data underwent method-specific centering and standardization, with outliers replaced by feature-specific means in training datasets. Testing occurred without these preprocessing steps. Model hyperparameters were optimized using a subset of testing data via 10-fold cross-validation. Models were trained on full datasets and tested on newly acquired spinach wash water samples. Synthetic Minority Oversampling Technique (SMOTE) and Adaptive Synthetic Sampling approach (ADASYN) achieved strong results, with SMOTE RF reaching an accuracy of 90.0%, sensitivity of 93.8%, specificity of 87.5%, and an area under the curve (AUC) of 98.2% (without data preprocessing) and ADASYN achieving 86.55% accuracy, 87.5% sensitivity, 83.3% specificity, and a 92.4% AUC. SMOTE and ADASYN significantly improved (p < 0.05) SVM and RF models, compared to their non-oversampled counterparts without preprocessing. Data preprocessing had a mixed impact, improving (p < 0.05) the accuracy and specificity of the BLR model but decreasing the accuracy and specificity (p < 0.05) of the SVM and RF models. The most influential physiochemical feature for E. coli detection in wash water was water conductivity, ranging from 7.9 to 196.2 µS. Following closely was water turbidity, ranging from 2.97 to 72.35 NTU within this study.

Metabolomics reveals the phytotoxicity mechanisms of foliar spinach exposed to bulk and nano sizes of PbCO3.

PbCO3 is an ancient raw material for Pb minerals and continues to pose potential risks to the environment and human health through mining and industrial processes. However, the specific effects of unintentional PbCO3 discharge on edible plants remain poorly understood. This study unravels how foliar application of PbCO3 induces phytotoxicity by potentially influencing leaf morphology, photosynthetic pigments, oxidative stress, and metabolic pathways related to energy regulation, cell damage, and antioxidant defense in Spinacia oleracea L. Additionally, it quantifies the resultant human health risks. Plants were foliarly exposed to PbCO3 nanoparticles (NPs) and bulk products (BPs), as well as Pb2+ at 0, 5, 10, 25, 50, and 100 mg·L-1 concentrations once a day for three weeks. The presence and localization of PbCO3 NPs inside the plant cells were confirmed by TEM-EDS analysis. The maximum accumulation of total Pb was recorded in the root (2947.77 mg·kg-1 DW for ion exposure), followed by the shoot (942.50 mg·kg-1 DW for NPs exposure). The results revealed that PbCO3 and Pb2+ exposure had size- and dose-dependent inhibitory effects on spinach length, biomass, and photosynthesis attributes, inducing impacts on the antioxidase activity of CAT, membrane permeability, and nutrient elements absorption and translocation. Pb2+ exhibited pronounced toxicity in morphology and chlorophyll; PbCO3 BP exposure accumulated the most lipid peroxidation products of MDA and H2O2; and PbCO3 NPs triggered the largest cell membrane damage. Furthermore, PbCO3 NPs at 10 and 100 mg·L-1 induced dose-dependent metabolic reprogramming in spinach leaves, disturbing the metabolic mechanisms related to amino acids, antioxidant defense, oxidative phosphorylation, fatty acid cycle, and the respiratory chain. The spinach showed a non-carcinogenic health risk hierarchy: Pb2+ > PbCO3 NPs > PbCO3 BPs, with children more vulnerable than adults. These findings enhance our understanding of PbCO3 particle effects on food security, emphasizing the need for further research to minimize their impact on human dietary health.

Development of packaging films based on UiO-66 MOF loaded melatonin with antioxidation functions for spinach preservation.

Spinach tends to deteriorate after harvest due to physiological metabolic activities. As a natural, pollution-free, and environmentally friendly preservative, melatonin (MT) can effectively maintain the quality of fruits and vegetables after harvest and delay senescence. To enhance the preservation effect of MT, this study developed antioxidant films using MT-loaded UiO-66 metal-organic framework (MOF) nanoparticles. This approach effectively extends the shelf life of spinach while preserving its quality. The underlying mechanism involves leveraging the microporous structure and stability of UiO-66 MOF. Experimental results obtained from the packaging films demonstrated significant improvements in both mechanical strength and antioxidant properties when UiO-66 was loaded with MT at a concentration of 0.20 mg/mL and combined with sodium alginate. Freshness preservation experiments also indicated the effective preservation effect of these films on spinach. In conclusion, the results of this study suggest that MT-loaded UiO-66 MOF is a promising active packaging material for spinach preservation.

Mechanism and synergistic effect of sulfadiazine (SDZ) and cadmium toxicity in spinach (Spinacia oleracea L.) and its alleviation through zinc fortification.

Cadmium (Cd) and antibiotic's tendency to accumulate in edible plant parts and fertile land is a worldwide issue. The combined effect of antibiotics and heavy metals on crops was analyzed, but not mitigation of their toxicity. This study investigated the potential of zinc oxide nanoparticles (ZnO NPs) to alleviate the SDZ and Cd toxicity (alone/combined) to promote spinach growth. Results revealed that the ZnO 200 mg L-1 spray decreased the malondialdehyde (MDA) 14%, hydrogen peroxide (H2O2) 13%, and electrolyte leakage (EL) 7%, and increased the superoxide dismutase (SOD) 8%, peroxidase (POD) 25%, catalase (CAT) 39% and ascorbate peroxidase (APX) 12% in spinach leaves under combined SDZ+Cd (25 mg Kg-1 +50 mg Kg-1) stress compared to ZnO 100 mg L-1 spray. Likewise, ZnO NPs 200 mg L-1 spray enhanced the zinc (Zn) 97%, iron (Fe) 86%, magnesium (Mg) 35%, manganese (Mn) 8%, and potassium (K) 23% in shoots under combined SDZ+Cd (25 mg Kg-1 +50 mg Kg-1) stress compared to ZnO 100 mg L-1 spray. Further, ZnO 200 mg L-1 spray reduced Cd uptake in roots by 9% and shoots 15% under combined SDZ+Cd (25 mg Kg-1 +50 mg Kg-1) stress compared to ZnO 100 mg L-1. Overall, ZnO NPs alleviated the SDZ and Cd toxicity and enhanced spinach growth in all treatments.