Ceramides contribute to the development of type 2 diabetes but it is uncertain whether they predict gestational diabetes (GDM). In this multicentre case–control study including 1040 women with GDM and 958 non-diabetic controls, early pregnancy  concentrations of four ceramides—Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1)—were determined by a validated mass-spectrometric method from biobanked serum samples. Traditional lipids including total cholesterol, LDL, HDL and triglycerides were measured. Logistic and linear regression and the LASSO logistic regression were used to analyse lipids and clinical risk factors in the prediction of GDM. The concentrations of four targeted ceramides and total cholesterol, LDL and triglycerides were higher and HDL was lower among women with subsequent GDM than among controls. After adjustments, Cer(d18:1/24:0), triglycerides and LDL were independent predictors of GDM, women in their highest quartile had 1.44-fold (95% CI 1.07–1.95), 2.17-fold (95% CI 1.57–3.00) and 1.63-fold (95% CI 1.19–2.24) odds for GDM when compared to their lowest quartiles, respectively. In the LASSO regression modelling ceramides did not appear to markedly improve the predictive performance for GDM alongside with clinical risk factors and triglycerides. However, their adverse alterations highlight the extent of metabolic disturbances involved in GDM. GDM predisposes both the woman and her child to major short- and long-term adverse health outcomes10. Among women with a history of GDM, up to half develop type 2 diabetes later in life6 and they also have a higher prevalence of several cardiometabolic risks, including hypertension, dyslipidaemia, metabolic syndrome and cardiovascular disease11.Gestational diabetes (GDM) is one of the most common pregnancy complications, affecting between 7 and 28% of pregnancies worldwide13. Among women with GDM, chronic insulin resistance is often present before conception, and hyperglycaemia develops when pancreatic compensatory mechanisms fail14. Besides being linked to hyperglycaemia, insulin resistance is also linked to alterations in lipid metabolism during pregnancy15. It is well documented that circulating traditional lipids, especially triglycerides, are associated with an increased risk of GDM16. However, the relationships between lipid metabolism and diabetic pathways are far more complex and require a better understanding than can be gained from measuring traditional lipids17.Physiological insulin resistance and alterations in lipid metabolism characterize a normal pregnancy22. Ceramides, the precursors of more complex sphingolipids, are formed from a sphingoid base attached to a fatty acid of varying length ranging from 14 to 34 carbon (C) atoms20. In particular, the oversupply of saturated fat leads to the accumulation of these biologically active metabolites in several tissues, such as the liver, adipose tissue and skeletal muscle. Ceramides can modify several intracellular signalling pathways, such as inhibiting insulin signalling by blocking protein kinase B (Akt), leading to decreased insulin-induced glucose uptake20.Ceramides (Cer) are sphingolipids which have a key role in the development of insulin resistance, type 2 diabetes and cardiovascular disease23. The value of three ceramides—Cer(d18:1/16:0), Cer(d18:1/18:0) and Cer(d18:1/24:1)—and their ratios to Cer(d18:1/24:0) in predicting cardiovascular risk, especially cardiovascular death has been previously demonstrated and validated for clinical use24. Further, ceramide species which contain C16:0  and C18:0 (stearic acid) have showed the strongest association with insulin resistance and incident type 2 diabetes25 and the ratio of ceramide stearic to palmitic acid [Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio] has found to be an independent predictor for incident type 2 diabetes18. Although ceramides and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio can be used to identify nonpregnant individuals at the greatest risk for type 2 diabetes, it is not clear whether they are useful in the prediction of GDM. In three studies, higher concentrations of early-pregnancy ceramide species containing C14:026, C18:128, and C18:028 were associated with subsequent GDM.Circulating ceramides are already elevated years before the onset of type 2 diabetes24 type 2 diabetes- and cardiovascular disease-associated serum ceramides Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio, as well as traditional lipids, measured in early pregnancy, are predictors of subsequent GDM.The aim of this large, case–control study to examine whether four previously validated30. Briefly, 1146 women with GDM and 1066 pregnant controls with no diabetes were recruited between 1 February 2009 and 31 December 2012 from delivery units in seven Finnish delivery hospitals, each serving its own geographic catchment area. Women with GDM were recruited at the delivery units as they entered to give birth, and the next-consenting mother with no diabetes giving birth at the same unit was recruited as a control. Women with multiple pregnancies or pregestational diabetes were excluded.This case–control study is a part of the clinical genetic arm of the Finnish Gestational Diabetes (FinnGeDi) study, which has been described in detail31. OGTT was performed already at the 12th–16th weeks of gestation on high-risk women , and it was repeated at the 24th–28th weeks of gestation if the first OGTT was normal. The cut-off values for the venous glucose concentrations were as follows: fasting ≥ 5.3 mmol/L; 1 h ≥ 10.0 mmol/L; and 2 h ≥ 8.6 mmol/L. At least one abnormal value was diagnostic for GDM. The GDM status of each participant was confirmed from the medical records.According to the Finnish National Current Care Guidelines, comprehensive screening of GDM by a 2 h 75 g oral glucose tolerance test (OGTT) was performed on all women at the 24th–28th weeks of gestation except for those with a very low risk for GDM Participants completed background questionnaires about their lifestyles and medical and family histories. Detailed data on pregnancy and delivery were collected from the hospital and maternal welfare clinic records and combined with individually linked register data obtained from the Finnish Medical Birth Register (FMBR).2) was calculated. Gestational weight gain was calculated as the difference between the pre-pregnancy weight and the weight at the last antenatal visit (≥ 35 weeks of gestation). Based on the questionnaire data, educational attainment was categorised as basic or less, upper secondary, lower-level tertiary or upper-level tertiary. Chronic hypertension was defined as systolic blood pressure ≥ 140 mmHg or diastolic blood pressure ≥ 90 mmHg measured repeatedly or the use of antihypertensive medication before 20 weeks of gestation, while gestational hypertension was defined when hypertension appeared after 20 weeks of gestation. Pre-eclampsia was considered when hypertension appeared after 20 weeks of gestation and was accompanied with proteinuria (≥ 300 mg/day or two ≥ 1 + readings on a dipstick test). Data on previous pregnancies, including prior GDM, were obtained from the questionnaire and the FMBR. The participants’ family history of type 2 diabetes was taken from the questionnaire.Data on maternal age at delivery, parity and smoking during pregnancy were obtained from the FMBR. Self-reported maternal height and pre-pregnancy weight were obtained from the maternal welfare clinic records, and BMI , a nationwide biobank containing leftover serum samples from routine the early pregnancy routine infectious disease screening. Therefore, fasting before sampling was not required. The samples were stored at − 25 °C in the Biobank Borealis of Northern Finland.n = 192: n = 124, no sample in the biobank; n = 68, sample was depleted) did not significantly differ from those of the samples included. Samples drawn after 20 weeks of pregnancy (n = 22) were excluded. The samples were drawn, on average, at 10.7 (SD 2.1) weeks of gestation. Finally, 1998 participants (1040 with GDM and 958 controls) were included in the analyses . The background characteristics of those participants with missing samples -labelled internal standards—D7-Cer(d18:1/16:0), D7-Cer(d18:1/18:0), D7-Cer(d18:1/24:0) and D7-Cer(d18:1/24:1)—and extracted in isopropanol:ethyl acetate . Quantification of the individual ceramides was performed in multiple-reaction-monitoring mode and assessed through calibration line samples composed of known amounts of synthetic Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:0) and Cer(d18:1/24:1) and corresponding 2H-labelled standards. The peak area ratio for each ceramide to its corresponding 2H-labelled form was calculated and plotted against the added concentration of ceramide, followed by linear regression analysis. Concentrations of ceramides are presented in µmol/L. The ratio of Cer(d18:1/18:0)/Cer(d18:1/16:0) was calculated. The NordLab clinical laboratory analysed traditional lipids  using a standard enzymatic assay on a Siemens Advia automatic biochemical analyser  in Oulu, Finland. Random first-trimester samples of pooled serum from the FMC were incorporated with the sample runs; these samples acted as internal controls, and the inter-assay coefficients of variation  were derived from these samples and intra-assay CVs were derived from the intra-assay quality control samples , Cer(d18:1/18:0), Cer(d18:1/24:0), Cer(d18:1/24:1)—were measured using a validated, targeted, liquid chromatography–tandem mass spectrometry assay by Zora Biosciences Oy in Espoo, Finland8:1/16:0,t-test for continuous variables  and the χ2 test for categorical variables (expressed as frequencies). The main outcome was GDM. We compared the means of traditional lipids, ceramides and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio using both linear and logistic regression. To estimate the association of each variable with GDM, mean differences and odds ratios (ORs) with 95% confidence intervals (CI) per SD and per quartile (Q2–Q4) were calculated using the lowest quartile (Q1) as a reference. Model 1 was unadjusted. In Model 2, the results were adjusted for pre-pregnancy BMI, age, parity (dichotomous variable: primipara/multipara) and gestational weeks at sampling. Model 3 was adjusted for Model 2 and educational attainment, a history of GDM, parental type 2 diabetes and delivery unit. Categorical variables were added as dummy-coded, with a separate dummy variable indicating missing values. The directed acyclic graph summarising the hypothetical causality between ceramides and traditional lipids and GDM, and potential confounding variables used in the regression analyses is shown in Supplemental Fig. Statistical analyses were performed using SPSS 28.0 and R software (version 4.2.1). The baseline characteristics of the study participants were described using the unpaired Student’s glmnet’); this regularization method is useful in selecting parsimonious predictive models, particularly when there is multicollinearity among covariates (as is the case with lipids in this study)32. Models with different sets of covariates were considered. First, clinical predictors for GDM , educational attainment and delivery unit, altogether 16 covariates, were considered. Then, all four ceramides, the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio and traditional lipids  were included. Furthermore, models including logarithmic, square, cubic and square root transformations of continuous covariates were considered.Last, to assess the relative contributions of covariates on the risk of GDM, a least absolute shrinkage and selection operator (LASSO) logistic regression analysis was constructed  value, and the average root mean squared error (RMSE) was recorded. The model corresponding to the optimal value  of lambda was selected. Furthermore, tenfold cross-validation was also applied to assess the out-of-sample prediction accuracy of various models. Here, the model was repeatedly fitted using 90% of the data, and the accuracy of the model predictions to the actual observed values (according to the AUC or RMSE criteria) for the remaining 10% of the data not used in the model fitting (holdout data) was evaluated.The power of the study was sufficient to identify small differences between the study groups. With a power of 0.80, a significance level of 0.05 and an effect size of d 0.13, we were able to detect a difference of 0.13 SD in lipids between women with GDM and the controls.The study was carried out according to the Declaration of Helsinki and approved by the Ethics Committee of Northern Ostrobothnia Hospital District (Reference Number 33/2008), the Finnish Institute for Health and Welfare and the scientific committee of the Northern Finland Biobank Borealis. All participants gave written informed consent after full explanation of the purpose and nature of all procedures used.The women with GDM were older, less often primiparous and had higher BMI and blood pressure than the controls Table . In the Overall, the early pregnancy concentrations of all four ceramides—Cer(d18:1/16:0), Cer(d18:1/18:0), Cerd18:1/24:0) and Cer(d18:1/24:1)—and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio were higher among women with GDM compared to women without GDM (Model 1)  showed 1.43-fold odds (95% CI 1.11–1.83), Cer(d18:1/18:0) showed 2.40-fold odds (95% CI 1.86–3.09), Cer(d18:1/24:0) showed 2.11-fold odds (95% CI 1.64–2.72), Cer(d18:1/24:1) showed 2.21-fold odds (95% CI 1.71–2.84) and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio showed 2.38-fold odds (95% CI 1.84–3.07) for GDM , Cer(d18:1/18:0), Cer(d18:1/24:0), Cer(d18:1/24:1) and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio, as well as triglycerides, LDL and total cholesterol were higher and HDL was lower among women who subsequently developed GDM. In logistic regression models with a single predictor, Cer(d18:1/24:0), triglycerides and LDL were independent predictors for GDM. In the LASSO regression modelling, in addition to clinical risk factors and triglycerides ceramides did not appear to markedly improve the predictive performance for GDM.22. Hypercaloric diet and obesity lead to excess delivery of fatty acids, which causes dysregulation of multiple lipid metabolic pathways and accumulation of numerous lipid subtypes such as ceramides20. Further, these changes in lipid metabolism promote insulin resistance, mitochondrial dysfunction, oxidative stress and inflammation17. Concentrations of several circulating ceramides elevate years before the onset of type 2 diabetes33.Ceramides play a lipotoxic role in the development of insulin resistance, type 2 diabetes and cardiovascular disease20. In nonpregnant populations, elevated levels of ceramides containing long acyl chains, such as C16:0 and C18:0, have shown the strongest association with insulin resistance and incident type 2 diabetes25. Instead, very long chains containing ceramides, such as C24:0, have been suggested as neutral or protective20; however, some studies have reported them to be associated with insulin resistance and type 2 diabetes35. When normoglycaemic women were studied 12 weeks after GDM pregnancy, the levels of C22:0 and C24:0 ceramide species were higher among those who develop type 2 diabetes in the long term33.Although the underlying mechanisms are not fully known, the length of the acyl chain of ceramide seems to play a role in the development of insulin resistance38. Maternal hyperlipidaemia is primarily aimed at securing fetal growth and development, especially in the third trimester38. Only a few studies have examined the associations of early pregnancy serum ceramide concentrations in subsequent GDM, with conflicting results40. Three previous studies, a prospective lipidomic study including 492 women with GDM26, a prospective cohort study including 53 women with GDM27 and a nested case–control study including 24328 women with GDM, reported that higher levels of circulating C14:026, C18:028, and C18:128 ceramide species in early pregnancy were associated with subsequent GDM.During pregnancy, the serum concentrations of several ceramides, as well as traditional lipids, are known to increase28, we also found that the early pregnancy concentrations of Cer(d18:1/18:0) were higher in women who developed GDM compared with those who did not, but the difference was mostly explained by their higher BMI and age. Further, the difference was attenuated by other clinical risk factors for GDM, such as a history of GDM and a family history of type 2 diabetes. Although Cer(d18:1/18:0) was selected for the LASSO model, it did not improve predictive performance alongside clinical risk factors and/or triglycerides. In line with our findings, in a recent lipidomic study of 336 women with GDM, C18:0 ceramide was not independently associated with GDM39. Instead, they detected by the LASSO regression 10 lipid biomarkers in three categories of lipid classes, including one upregulated glycerolipid, five glycerophospholipids and four downregulated sphingolipids. Furthermore, in two lipidomics studies of 10740 and 10036 women with GDM, some di- and triacylglycerides were independent biomarkers for GDM, but ceramides were not40.In line with previous findings18, in this study among pregnant women, it did not improve the predictive performance of GDM when clinical risk factors, especially pre-pregnancy BMI or triglycerides, were considered. Further, we found Cer(d18:1/24:0) to be an independent predictor for GDM; in contrast, previous studies with smaller sample sizes found either an inverse association41 or no association between C24:0 and GDM27. Although Cer(d18:1/24:0) was positively related to GDM, it did not improve the prediction of GDM when clinical risk factors and triglycerides were considered.Although the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio has been shown to be an independent predictor of type 2 diabetesPossible explanations for discordant results may be differences in sample size and settings of studies, diagnostic criteria of GDM, varying methods of determining ceramides and differences in study populations and analysing methods, including adjustments for covariates, especially where pre-pregnancy BMI plays an important role.24, were measured during early pregnancy in a large group of pregnant women in this well-defined case–control setting. This was the first study assessing the early pregnancy levels of these ceramides and the Cer(d18:1/18:0)/Cer(d18:1/16:0) ratio together with traditional lipids and evaluating their roles as predictors for subsequent GDM. The LASSO regression, also previously applied in several lipidomic studies assessing circulating lipids in early pregnancy with subsequent GDM42, was selected as an efficient method to create a parsimonious predictive model in the presence of multicollinear predictors. The GDM status of each participant was confirmed from the medical records, and several potential confounders were considered in the analyses. The study provides reference data for ceramide lipids among pregnant women in relation to GDM status.This study has several strengths. Four ceramides, previously validated in nonpregnant populations43. Secondly, the majority of the participants were of Finnish ancestry, which may limit the generalisability of the findings. Thirdly, the quantifications of other ceramides, diacyl- or triacylglycerols, or lipidomic analyses could have brought a broader perspective to this subject but they were not possible to realise within this study. Finally, the results could not be validated in an external cohort; to control this limitation, the LASSO regression and cross-validation were performed.The study also has some limitations. Firstly, serum samples were taken at non-fasting state, which may have a minor effect on triglyceride levelsFuture studies with independent external validation are needed to confirm the findings of this study. Further, it would be important to study whether these early pregnancy alterations in lipid metabolism are related to the long-term metabolic health and development of type 2 diabetes, and whether the ceramide profile varies depending on the stage of the diabetic cascade.The early pregnancy levels of ceramides and traditional lipids were higher among women who developed GDM compared to those who did not. Cer(d18:1/24:0), triglycerides and LDL were found to be independent predictors of GDM. Clinical risk factors played a dominant role in predicting GDM and after combined with triglycerides, ceramides did not markedly improve the predictive performance for GDM. However, adverse alterations in lipids reflects the clustering of metabolic risk factors related to GDM.Supplementary Information.


Those genotypes included Giza-168, Giza-171, Misr-1, Misr-3, Shandweel-1, Sids-1, Sids-12, and Sids-14. The study included six treatments as follows: regular irrigation with 100% Field Capacity (FC) at a temperature of 23 ± 3 °C (T1), drought stress with 60% FC (T2), heat stress of 38 °C for 5 h·day−1 (T3), foliar spray of 10 mg·L−1 of SeNPs only (T4), a combination of drought stress with foliar spray of 10 mg·L−1 of SeNPs (T5), and heat stress with foliar spray of 10 mg·L−1 of SeNPs (T6). The experiment continued for 31 days. Foliar application of SeNPs improved the plant growth, morpho-physiological and biochemical responses, and expression of stress-responsive genes in wheat (T. aestivum L.) seedlings. Overall, morpho-physiological traits such as plant height (PH), shoot fresh weight (SFW), shoot dry weight (SDW), root fresh weight (RFW), and root dry weight (RDW) of wheat genotypes grown under different conditions ranged from 25.37–51.51 cm, 3.29–5.15 g, 0.50–1.97 g, 0.72–4.21 g, and 0.11–1.23 g, respectively. From the morpho-physiological perspective, drought stress had a greater detrimental impact on wheat plants than heat stress, whereas heat stress significantly impacted the expression of stress-responsive genes. Stress responses to drought and heat varied between wheat genotypes, suggesting that different genotypes are more resilient to stress. Exogenous spraying of 10 mg·L−1 of SeNPs improved the photosynthetic pigments, photosynthetic rate, gas exchange, and transpiration rate of wheat plants and enhanced drought and heat tolerance by increasing the activity of antioxidant enzymes including catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD) and the expression level of stress-responsive genes. Our results showed that spraying wheat seedlings with 10 mg·L−1 of SeNPs enhanced SOD activity for all genotypes as compared to the control, with the Sids-12 genotype having the highest value (196.43 U·mg−1 FW·min−1) and the Giza-168 genotype having the lowest (152.30 U·mg−1 FW·min−1). The expression of PIP1, LEA-1, HSP70, and HSP90 stress-responsive genes was more significant in tolerant genotypes (Giza-171 and Giza-168) than in sensitive ones (Misr-1 and Misr-3) in response to drought and heat stresses. Under stress conditions, the shoot and root fresh weights, photosynthetic pigment content, stomatal conductance (SC), and transpiration rate (TR) were positively correlated with plant height (PH), while root and shoot dry weights, malondialdehyde (MDA), proline, hydrogen peroxide (H2O2), and APX were negatively correlated. Multivariate analysis and biplot results revealed that genotypes Giza-168, Giza-171, Sids-12, and Sids-14 performed well in both stress situations and were classified as stress-tolerant genotypes. These best genotypes may be employed in future breeding projects as tools to face climate change. This study concluded that various physio-biochemicals and gene expression attributes under drought and heat stress could be modulated by foliar application of SeNPs in wheat genotypes, potentially alleviating the adverse effects of drought and heat stress.In this study, the role of selenium nanoparticles (SeNPs, 10 mg·L Triticum aestivum L.) is a staple crop. The global wheat industry faces numerous challenges, including the convergence of genetic resources in behavior and productivity, biotic and abiotic stress, soil poverty, and an increase in global consumption because of an increase in the world population, global supply chains, and a monopoly on fertilizers for better agricultural practices. According to [https://www.fas.usda.gov/data/egypt-grain-and-feed-update-5, accessed on 20 January 2023).Globally, wheat  and selenite (SeO3−2). Size reduction will also boost the activity of nanoparticles and make them more efficient [Physiological and biochemical changes occur in wheat plants due to drought stress, including hormonal imbalance, ethylene production, and reduced photosynthesis, because drought affects cell and tissue structure and important processes at all stages of growth . As a refficient .According to the best of our knowledge, there are no reports available about the drought or heat tolerance status at the seedling stage of the investigated wheat genotypes. Therefore, the current study aimed to screen the selected wheat genotypes for drought and/or heat tolerance and to investigate the impacts of chemically synthesized SeNPs on the physiology, biochemistry, and expression level of stress-responsive genes induced by drought and heat stress in the eight wheat genotypes chosen for this investigation. This could help identify the characteristics that influence the drought and heat tolerance of wheat plants for use in breeding programs in the future.T. aestivum L.) genotypes: Giza-168, Giza-171, Misr-1, Misr-3, Shandweel-1, Sids-1, Sids-12, and Sids-14 were obtained from the Wheat Research Department, Field Crops Research Institute, Agricultural Research Center (ARC), Ministry of Agriculture, Egypt. These genotypes are the most widely used by Egyptian farmers and in the local Egyptian markets. The names of these genotypes and pedigrees are listed in The SeNPs used in this study were obtained from Sigma-Aldrich  in physical dispersion form, with average particle sizes ranging from 10 to 100 nm and a concentration of 0.15% of selenium in water. Eight bread wheat (The form and size of selenium nanoparticles were investigated using transmission electron microscopy (TEM) . An aqueous suspension containing the SeNPs was dispersed ultrasonically, and the sample was prepared by placing a drop of the solution on a copper grid coated with carbon and drying it under an IR lamp. Micrographs were obtained using a TEM operated at an accelerating voltage of 80 kV. The related size distribution pattern of SeNPs was plotted by manually counting 200 individual particles from different TEM images.−1–4000 cm−1.Fourier transform infrared spectroscopy analysis was done to characterize the functional groups on the surface of SeNPs. A powder sample of SeNPs was used for FTIR spectroscopy measurements. Powder and potassium bromide (KBr) were mixed in a 1:100 ratio, and a Shimadzu FTIR  apparatus was used to record the spectra. The FTIR spectrum was obtained with wave numbers between 500 cmA pot experiment was conducted at the greenhouse of Scientific Research City and Technological Applications (SRTA-City), New Borg El Arab City, Egypt, to examine the impact of SeNPs on the morpho-physiological, biochemical, and molecular responses of eight bread wheat genotypes growing under drought and heat stress conditions. In plastic pots (10 cm diameter × 12 cm depth), filled with soil, clay, and compost (4:1), and adding 10 g of N, K, and P (10-5-5) nutrients. After wheat seed sterilization, we sowed 20 homogenous, healthy seeds in each pot and submerged the pots with ¼ strength Hoagland nutrient-supplemented irrigation water  every ot−1 of SeNPs with regular irrigation (100% FC) and temperature of 23/17 ± 3 °C; T5, drought stress (60% FC) + 10 mg·L−1 of SeNPs; and T6, heat stress (38 °C) + 10 mg·L−1 of SeNPs. All those treatment conditions were examined in pot experiments with three replicates for each treatment, which continued until the end of the experiment. After 1 week (7 DAS), the germinated wheat seedlings were reduced to 8 wheat seedlings per pot (thinning) to have homogeneity among all pots and continued germination for another 2 weeks (21 DAS). During these 2 weeks, the drought stress treatments (T2 and T5) were with moderate drought (60% FC) by irrigation every 3 days, and the heat stress treatments (T3 and T6) were at 38 °C (5 h·day−1). Finally, at 21 DAS, all pots were moved to the greenhouse standard conditions (regular irrigation (100% FC) and 23/17 ± 3 °C), and an exogenous application of 10 mg·L−1 of SeNPs (20 mL·pot−1) was applied to T4, T5, and T6 at 3-day intervals for 10 days. Distilled water was used for the control treatment (T1). At the end of the experiment (31 DAS), samples were collected for further analysis. SeNPs concentration (10 mg·L−1) was selected in this study according to previous studies that recommended using 30 mg·L−1 or less of SeNPs. The SeNPs at a concentration of 30 mg·L−1 or less were found to be the most suitable concentrations that enhanced the morphological, physiological, biochemical, and antioxidant parameters under different biotic and abiotic stresses [Spirulina platensis polysaccharides (SPs) as stabilizers to prepare SPs-SeNPs and reported that the median toxic concentration (TC50) of SPs-SeNPs was approximately 22,000 μg·L−1. Three replicates for each treatment were established (The greenhouse photoperiod of 13/11 h (light/dark), air relative humidity of 65 ± 5%, and (day/night) temperature were maintained at 23/17 ± 3 °C throughout the pot experiments other than the heat stress condition. The treatments were as follows: T1, control ) at (day/night) temperature of 23/17 ± 3 °C; T2, drought stress (60% FC); T3, heat stress (38 °C) with regular irrigation (100% FC); T4, 10 mg·Lstresses . Additioablished .2) per plant were measured at the final harvesting date.At the end of the experiment (31 DAS), plant samples from three separated pots in each treatment were harvested. To estimate growth and physiological parameters, three wheat plants from each treatment were taken and washed with distilled water to remove soil and adhering particles. A measuring scale immediately assessed plant height (PH) (cm) after harvest. Calculations were made for shoot fresh weight (SFW) and root fresh weight (RFW). To clean the root for mass analysis, the root system was held at the plant base and gently shaken free of bulk soil. The root was then transferred to a clean sheet of paper, and the soil that was not rigidly attached to the roots was gently brushed off with a soft brush. The soil that remained attached to roots after this treatment was considered rhizosheath soil. After that, the roots were washed over a 0.5-mm mesh, blotted, and weighed to obtain the fresh root mass. Additionally, after oven-drying at 65 °C for 48 h, the shoot dry weight (SDW) and root dry weight (RDW) were determined. The number of leaves per plant and leaf area . The net photosynthetic rate (PN), stomatal conductance (SC), and transpiration rate (TR) were measured using an infrared gas analyzer system . These parameters were measured in the middle region of the uppermost fully expanded leaves of treated and non-treated wheat plants on a bright sunny day between 9:30 and 11:30 a.m.According to Arnon, chlorophyll content (CHLO) in the uppermost fully expanded leaves was determined spectrophotometrically . A leaf Electrolyte leakage (EL) was measured by boiling fresh leaf discs in 10 mL of deionized water, and electrical conductivity (EC1) was measured. After that, tubes were heated at 55 °C for 30 min, and electrical conductivity (EC2) was measured again. Again, after boiling the tissue for 10 min at 100 °C, electrical conductivity (EC3) was recorded . The calThe method of Sun et al.  was usedOn fully expanded leaves of physiological activity-controlled wheat plants and wheat plants under stress, leaf water potential (Ψ) was measured. Then, based on the formula described by Campos et al. , the wat2O2 levels were measured according to the previously described method [g for 15 min. Then, in a reaction mixture of 0.5 mL enzyme supernatants, 0.5 mL phosphate buffer (pH 7.0), and 1 mM potassium iodide, the absorbance of the mixture was measured at 390 nm against H2O2 as a standard.The Hd method . BrieflyT. aestivum fresh leaves. Briefly, 0.5 g of leaf samples were homogenized in 10 mL of ethanol and then centrifuged for 10 min at 10,000× g. An amount of 1 milliliter of the extract was added to 2 milliliters of 0.65% thiobarbituric acid (TBA) in 20% TCA. After boiling for 30 min, the mixture was rapidly cooled. Following centrifugation of the samples at 10,000× g for 5 min. The MDA content was determined by comparing non-specific absorption at 600 and 532 nm.A method described by Heath and Packer  was usedg, 2 mL of the supernatant was added to 2 mL glacial acetic acid and 2 mL of the ninhydrin reagent, and the reaction was incubated at 100 °C for 1 h. Afterward, tubes were kept in an ice bath, and toluene was used to separate the proline. An optical density of 520 nm was then determined [Proline was determined by extracting 500 mg of dried powdered leaf tissue in 3% sulphosalicylic acid. Following centrifugation of the extract for 20 min at 3000× g for 20 min at 4 °C. The activity of superoxide dismutase (SOD) was determined by the reduction of nitroblue tetrazolium (NBT). The SOD was measured at 560 nm in an assay mix containing sodium phosphate buffer , 100 µL EDTA, L-methionine, 75 µM NBT, riboflavin, and 100 µL enzyme extract [Using a prechilled pestle and mortar, we homogenized 1 g of fresh leaf tissue in a chilled 50 mM phosphate buffer (pH 7.0) with 1% polyvinyl pyrrolidone and 1 mM EDTA to extract the antioxidant enzymes. To obtain enzyme sources, the homogenate was centrifuged at 15,000×  extract . One uni2O2. The reaction mixture contained 25 mM phosphate buffer (pH 7.0), 10 mM H2O2, and 0.1 mL enzyme extract.To determine ascorbate peroxidase (APX) activity, we monitored the absorption change in the rate of ascorbate oxidation at 290 nm for 3 min in a reaction mixture containing potassium phosphate buffer (pH 7.0), 0.5 mM ascorbic acid, hydrogen peroxide, and enzyme extract in a 1 mL reaction volume according to Nakano and Asada . To dete−1), 2.5 μL 5× buffer, 2.5 μL MgCl2, 2.5 μL 2.5 mM dNTPs, 4 μL from oligo (dT), 0.2 μL (5 Unit·μL−1) reverse transcriptase , and 2.5 μL RNA. The reverse transcription PCR (RT-PCR) amplification reaction was conducted in a thermal cycler PCR, programmed at 42 °C for 1 h and 72 °C for 20 min.Total RNA was isolated from a 0.5 g sample of wheat plants of all treatments by using the Plant RNA Kit (Sigma-Aldrich) according to the manufacturer’s instructions. The purified RNA was quantitated spectrophotometrically and analyzed on a 1.5% agarose gel. For each sample, 10 μg total RNA was treated with DNAse RNAse-free (Fermentas), 5 μg of which was reverse transcribed in a reaction mixture consisting of oligo dT primer . The relative gene expression was determined using the 2−ΔΔCt method [Quantitative Real-time PCR (QPCR) was conducted on 1 μL diluted cDNA in triplicate using real-time analysis using the Rotor-Gene™ 6000 system . The primer sequences used in QPCR are listed in t method .p ≤ 0.05 using SPSS 16.0 . Data were collected in a randomized complete block design with three replications. Histograms and violin plots were plotted using GraphPad Prism 9 . Normality was assessed using Shapiro–Wilk normality testing at the 0.05 level. Correlation coefficient matrices adapted to compute normalized mean values for generating scatter plots and heatmaps of the morpho-physiological parameters among studied genotypes. Principal component analysis (PCA-biplot) is a multivariate approach that examines the distribution of eight wheat genotypes based on data matrices comprising many associated quantitative dependent variables. Additionally, PCA-biplots and the score plots PC1 and PC2 were conducted of all data under different conditions. The PCA-biplot analysis was performed using JMP®, Version 16 .For morpho-physiological and molecular parameters among the genotypes, all data obtained were subjected to two-factor (treatment × genotypes) analysis of variance (ANOVA) using the general linear model, and the mean differences were compared using Tukey’s HSD test at The morphology of the SeNPs was observed using TEM, as shown in −1. A small band at 2345 cm−1 is associated with nitro compounds (N-O asymmetric stretch) present in the mixture, and a strong band at 1652 cm−1 with the C=O stretching vibrations. The band at 1571 cm−1 was due to N–O asymmetric stretch nitro compounds. Two sharp peaks at 1126 and 991 cm−1 were associated with the bending vibrations of the CH2 groups. The 933 cm−1 represented the stretching of C-N bonds in amines. A band at 916 cm−1 resulted from carboxylic acid’s O–H bend. As a result of C–X stretching in alkyl halides, there were bands at 814 and 694 cm−1. According to these results, various functional groups related to surface proteins in SeNPs may be responsible for their reduction and stabilization.The FTIR analysis of SeNPs is provided in −1 of SeNPs (T4), drought stress + 10 mg·L−1 of SeNPs (T5), and heat stress + 10 mg·L−1 of SeNPs (T6) are presented in As shown in The performance of eight wheat genotypes on morphological traits grown under control, drought, and heat stress conditions without and with SeNPs foliar application was scored . The res−1 of SeNPs significantly reduced the negative impacts of drought and heat stress on seedling growth parameters of wheat genotypes, where Sids-1 and Sids-12 recorded the highest responses (51.51 cm and 47.31 cm) to SeNPs for PH under drought and heat stress, respectively (Foliar application of 10 mg Lectively A. In theectively .−1. FW and 5.52 μmol (CO2) m−2·s−1), respectively, under drought stress, while it was at 2.90 mg·g−1. FW and 5.72 μmol (CO2) m−2·s−1, respectively, under heat stress, as compared to the control wheat plants (−1 FW and 4.80 μmol (CO2) m−2·s−1), respectively, under drought stress were reported in the Giza-171 genotype, while under heat stress they were 1.84 mg·g−1 FW and 5.02 μmol (CO2) m−2·s−1, respectively, as compared to the control. Under regular irrigation conditions (100% FC) and foliar application of 10 mg·L−1 of SeNPs, leaf chlorophyll content and photosynthetic rate percentage were increased in all wheat genotypes. The maximum percentage of chlorophyll content (4.12 mg·g−1 FW) and photosynthetic rate (6.98 μmol (CO2) m−2·s−1) were recorded in Sids-12 genotypes, respectively, due to foliar application of SeNPs. Under drought and heat stress, spraying with SeNPs (10 mg·L−1) increased leaf chlorophyll content to the maximum percentage (3.35 and 3.45 mg·g−1 FW) in the Sids-12 and Sids-1 genotypes, respectively (2) m−2·s−1) were recorded in the Sids-12 genotype sprayed with SeNPs and drought or heat stress, respectively, compared to wheat plants treated with drought or heat stress only (The effect of SeNPs on leaf chlorophyll content (CHLO) and photosynthetic rate (PN) for eight wheat genotypes under drought and heat stress conditions was investigated. As shown in t plants A,B. Howeectively A,B. Meaness only A,B.The indicators of stomatal conductance (SC) and transpiration rate (TR) of wheat seedlings were significantly reduced when wheat plants were treated with drought and heat stress either alone or in combination with SeNPs compared to the control wheat plants .2O·m−2·s−2) of SC and TR for wheat seedlings treated by drought stress alone were recorded in Misr-1 and Giza-168 genotypes, respectively, compared to the control wheat plants, demonstrating their tolerance/resistance to drought stress. However, the highest decreasing values (11.26 and 0.60 mmol H2O·m−2·s−2) were found in the Sids-1 and Sids-12 genotypes, respectively. Regarding heat stress, the lowest decreasing values (18.01 and 0.74 mmol H2O·m−2·s−2) of SC and TR appeared in the Sids-12 genotype, reflecting their resistance to heat stress, and the highest decreasing values (16.51 and 0.55 mmol H2O·m−2·s−2) observed in the Sids-12 and Giza-168 genotypes treated by heat stress alone compared to the control wheat plants  and TR (0.81 mmol H2O·m−2·s−2) was recorded in the Sids-12 genotype. On the other hand, treatment with SeNPs (10 mg·L−1) in drought stress decreased the SC percentage in all wheat genotypes except Sids-12, where the SC percentage increased by 14.16% compared to drought stress control. However, the SC percentage increased in all studied wheat genotypes treated by heat stress with SeNPs except the Shandaweel-1 genotype, and the maximum percentage value of SC (20.30 mmol H2O·m−2·s−2) was reported in the Sids-12 genotype compared to heat stress control (Under the full-irrigation condition (100% FC), the application of SeNPs increased stomatal conductance and transpiration rate in all wheat genotypes, where the maximum percentage value of SC  and leaf water potential (LWP) as compared to individual heat stress and the control . HoweverThe highest increases in LWP (1.15 and 1.11 MPa) were observed in the Shandaweel-1 genotype under drought and heat stresses, respectively; however, the lowest increases in LWP (0.15 and 0.92 MPa) were found in the Misr-3 and Sids-12 genotypes as compared to the control wheat plants F. Notabl−1 of SeNPs resembled its content compared to spraying with water, and the greatest value was observed in Shandaweel-1 and the lowest one was noticed in Giza-168 compared to other genotypes  in the Giza-168 wheat genotype compared with the control.The proline concentrations in the leaves of wheat plants were significantly affected under drought and heat stress . The lea−1 of SeNPs enhanced SOD activity for all genotypes as compared to spraying with water, with Sids-12 having the highest value and Giza-168 having the lowest value.Compared to the control treatment, the imposed stressors with and without SeNPs spraying increased SOD activity. SOD activity rose considerably with stress exposure, with the maximum SOD activity recorded under drought and heat stresses in the Sids-14 wheat genotype. In the control treatment, the Giza-168 genotype had the lowest SOD activity . It was −1 of SeNPs did not show a significant difference in CAT activity.Drought raised ascorbic acid peroxidase (APX) activity more than heat stress in all wheat genotypes . The hig2O2, proline, APX, and CAT under drought stress. Likewise, SFW and RFW positively correlated with CHLO, PN, TR, and SOD, whereas they were negatively correlated with SDW, RDW, SC, EL, MDA, H2O2, proline, APX, and CAT , aquaporin (PIP1), heat shock protein 70 (Hsp70), and heat shock protein 90 (Hsp90) genes. They were conducted on 8 wheat genotype seedlings at 31 DAS under drought and heat stresses in the presence or absence of SeNPs  compared to the control.Under heat stress with and without SeNPs, both Giza-168 and Giza-171 showed the highest upregulation of gulation C,D. In dA PCA-biplot was used to analyze the average data of morpho-physiological traits and genomic constitution. PCA reflects the significance of the most significant contributor to total variation along each differentiation axis. PCA revealed similar groups, correlating the results of the interrelationship among all measured morpho-physiological and molecular parameters under different conditions .2O2 are the most extended vectors, and the small angle between these vectors proves a significant and strong positive correlation between these traits in cluster II. On the other side, SOD and APX traits had a strong positive correlation with each other but had an extremely negative correlation with proline, LWP, and H2O2. Moreover, the vector for MDA showed the strong discrimination power of the Shandweel-1 genotype among the other wheat genotypes (Under regular irrigation (control), variation in the eight-wheat genotype described by the first two PCs per cluster ranged from 19.5 to 51.5% of the total variation. In the system of the first two components, the length of the vector and cosine of the angle were used to discriminate wheat genotypes A. The eienotypes A.LEA-1, SDW, EL, and SDW. Additionally, HSP70 and RDW had a strong positive correlation with each other, whereas HSP70 and SOD had larger cosines of the angle with which they were not likely to be correlated. Cluster II had four genotypes , with frequent vectors such as HSP90, PH, TR, RFW, and SFW that had a strong positive correlation with each other. LWP and CAT were the most frequent vectors to differentiate cluster III (Sids-14 and Shandweel-1) genotypes from the others  with strong vectors such as CHLO, PN, SC, RFW, and SFW. EL and PH vectors had strong discrimination power between clusters II and III, which had three genotypes . The most frequent vector, SOD, which differentiates within cluster III genotypes, had a negative correlation with RDW , which differentiated from others by MDA, PIP1, APX, and proline  as described in Under drought and SeNPs conditions, PCA1 reported 58.1% of the total variation, while PCA2 described a 13.1% E. The thHSP70, HSP90, PIP1, LEA-1, H2O2, EL, APX, and MDA vectors; cluster II had four genotypes  with SC, TR, PH, CHLO, and RFW vectors; and cluster III had two genotypes (Sids-14 and Shandweel-1) with LWP, CAT, and SOD vectors, which strongly positive correlated as observed in Under heat and SeNPs conditions, PCA1 reported 59.4%, while PCA2 described 15.9% of the total variation F. The thAccording to current climate estimates, drought and heat waves are expected to become more prevalent and extreme in the future . Every d−1) improved all seedling development metrics  in wheat types when compared to negative control circumstances (drought and heat stress) and normal watering (100% FC). Seedling metrics improved more pronouncedly in Giza-168 and Sids-12 genotypes treated with 10 mg·L−1 of SeNPs. Our findings support the findings of Shoeibi et al. [−1 of SeNPs is more effective in boosting the development of wheat under drought conditions compared to the control.A range of strategies is being utilized to increase plant abiotic stress tolerance, including NPs, which are expected to boost efficiency under stressful circumstances. NPs improved germination and seedling development, physiological and biochemical activities such as photosynthesis and nitrogen metabolism, chlorophyll quality, antioxidant enzyme activities, carbohydrate and protein content, and positive gene expression, implying their potential use in crop enhancement . Accordii et al. , who demi et al. . Treatmei et al. . Furtheri et al. . Furtheri et al.  discoverShoot length, root length, and fresh weight are considered essential criteria for choosing drought-tolerant wheat genotypes . In wheaThe scatter plot and heatmap correlation matrix between morpho-physiological and biochemical parameters of eight wheat genotypes were used in this investigation. Plant height, RFW, SFW, RDW, and SDW were among the seedling morphological features evaluated. Understanding the relationship between these parameters was critical for improving the effectiveness of wheat drought tolerance breeding . A corre2 fixation, resulting in decreased photosynthesis and chlorophyll concentrations. Photosynthesis is also particularly sensitive under heat and drought circumstances, as the ratio and amount of chlorophyll (a and b) and carotenoid decreased with rising heat and drought severity [Drought directly affects wheat on all yielding and yield-related traits, as the yield decline due to persistent drought stress CD, 83.60%) was much greater than that due to terminal drought stress  . A tempeseverity . Furtherseverity . In this% was mucseverity ,61.Plant tissue temperature is sustained via water intake and transpiration, which stabilizes the water content of the tissue. However, when there is water scarcity, a temperature rise might be fatal . Leaf re−1 concentration, the Se/SiO2 NPs improve plant properties such as relative water content (RWC), membrane stability index, and water usage efficacy [Our results supported the findings of Bezabeh et al. , who disefficacy .Heat, salt, and drought stress all induce the creation of reactive oxygen species (ROS), which cause oxidative damage in the plant and, eventually, death. Under heat stress, ROS accumulate, causing severe oxidative damage to the plant and impeding growth and development . To comb2O2 is a hazardous molecule that harms cells by causing lipid peroxidation and membrane damage [2O2 accumulation in wheat plants, resulting in greater H2O2 accumulation in wheat plants subjected to both conditions. As previously stated, the drought and heat stress-induced rise in H2O2 led to a significant increase in leaf electrolyte leakage EL and MDA. The use of nano-selenium affects and improves germination and is a strong seed, as the coefficients of stem length, root length, wet weight, and dry weight were improved by 22.8, 24.9, 19.2, and 20%, respectively, over untreated controls [A high MDA concentration in this study implies membrane lipid peroxidation, which might indicate the amount of damage under unfavorable conditions. MDA content increased much faster in Giza-168 and Giza-171 seedling leaves than in other wheat genotypes during drought and heat stress, indicating their susceptibility to both stressors. Hcontrols .−1 of SeNPs, proline concentrations decreased significantly, perhaps mitigating the deleterious effects of drought and heat stress. According to Amoah et al. [2O2 accumulation, MDA, and proline content was 64.20%, 25.67%, 54.49%, and 14.49%, respectively, while the minimum increase in these parameters was 43.01%, 5.61%, 25.81%, and 5.6%, respectively, in wheat genotypes under drought stress. This is consistent with earlier research that has found even larger increases in heat-stressed wheat plants. In addition, Iqbal et al. [−1) reduced the proline content (4%) of wheat plants’ resistance to heat stress. Although osmolytes increased in some AgNPs (50 mg·L−1) treated with heat stress, the total rise was less significant when compared to heat stress application alone. Osmolytes, such as proline, play a critical role in providing resistance against abiotic stress  or SeNPs (T4) had reduced enzyme activity. Previously, increased POD and APX activities were found to operate as a mechanism of plant drought resistance [Antioxidant enzymes such as SOD, CAT, and APX are all key antioxidant enzymes in the wheat genotypes in this study. SOD activity increased much more than CAT and APX in all sensitive and tolerant genotypes in response to stress. When wheat plants were treated with 10 mg·Lsistance . Our finsistance  reportedsistance .2 and H2O2, which can lead to lipid peroxidation [2O2, and proline correlations after SeNPs administration to drought stressed wheat genotypes, with MDA becoming negatively associated with H2O2 and showing an increasingly positive correlation with proline, while H2O2 became negatively correlated to proline, APX, and CAT. After applying SeNPs to heat-challenged wheat genotypes, the MDA, H2O2, and proline interacted favorably with SOD, while SOD became positively connected with APX and CAT. When SeNPs are applied to wheat plants, they may cause acclimation reactions that lead to abiotic stress tolerance by up-regulating antioxidant enzymes. Although not yet recognized, seleniferous peptides and selenoamino acids are predicted to either directly neutralize ROS or do so through the production or modulation of other antioxidants in wheat plants [2 NPs are known to increase drought tolerance in stressed plants by boosting the activity of antioxidant enzymes such as GPX, SOD, APX, and CAT, as well as lowering lipid peroxidation and H2O2 concentrations [Drought decreases or eliminates chlorophyll concentration, which generates ROS such as Oxidation . Droughtxidation . There wt plants . Se/SiO2trations .Non-enzymatic antioxidant systems comprise low-molecular-weight compounds such as vitamins (vitamins C and E), β-carotene, uric acid, glutathione, ascorbic acid, carotenoids, phenolic compounds, α-tocopherols, alkaloids, flavonoids, carbohydrates, oils, anthocyanin, proline, glycine-betaine, non-protein amino acids, and hormones .Hsp70 and Hsp90 genes was elevated in this study when compared to the response to elevated temperature. Our findings are consistent with those of Bi et al. [TaHsfA2e-5D was expressed consistently in numerous wheat tissues, most notably roots during the reproductive stage. Heat, cold, drought, high salinity, and various phytohormones increased the expression of TaHsfA2e-5D in wheat seedlings.Heat stress significantly influenced gene expression, but there were only minor variations in methylation patterns in a genome-wide analysis of DNA methylation in wheat. However, methylation was linked to minor alterations in gene expression of key genes during heat stress in certain circumstances . These fi et al. , who detHVA1 (AT3G15670.1), a barley member of this protein group, has been demonstrated to be positively correlated with drought tolerance in barley and other plant species [LEA gene reported to be drought-responsive is WSI18 (AT3G15670.1) [LEA genes. Transcription factors alter shape when a wheat plant encounters water loss or scarcity. When exposed to drought stress, the LEA protein reacts to helicase, glucose, or rubisco [LEA genes, whereas other plant species include around 15 dehydrin genes and 100 LEA genes [MIP family, are often reduced in abundance, so helping to limit water loss [PIP1 and LEA-1 genes were considerably increased in response to drought stress, particularly PIP1 genes. Drought and heat stress induced the expression pattern of most LEA-1 genes under different conditions as evaluated by qRT-PCR analysis. The tolerant genotypes can modify genes under drought stress more strongly than the more vulnerable upland cotton genotypes [A number of the ABA-inducible genes were up-regulated by the stress encoded LEA proteins.  species . Another15670.1) . Wheat i rubisco . Wheat cEA genes . FurtherEA genes . Howeverter loss . In our enotypes .LEA-1, and HSP90. The most common vectors under heat stress were RFW, SFW, CHLO, PN, SC, MDA, H2O2, proline, APX, SOD, HSP90, and PIP1. Our PCA results were comparable to those published by wheat researchers [Principal component analysis (PCA) is a multivariate statistical tool used to examine and simplify complicated and enormous datasets. Only PCAs with eigenvalues greater than one were considered significant. During heat conditions, PCA1 contributed 65.1% to the total variance and PCA2 contributed 13.1%. Xhulaj et al.  discoverearchers . In thisAs a last point, selenium nanoparticles (SeNPs) increase the fraction of organic Se in wheat grain, which will improve the human body’s ability to absorb Se . In this−1 of SeNPs fortifies wheat crops against stress and improves morpho-physiological, biochemical, and molecular characteristics. It was found that four stress-responsive genes were significantly induced, suggesting that the ability of wheat plants to acclimate and induce antioxidant defenses may play a part in determining genotypic differences in drought and heat tolerance. Foliar application of SeNPs on wheat plants under stress enhances plant growth and maintains better water relations, reduces membrane damage, improves photosynthetic activity, and shows differentiation in gene expression trends. Outperforming drought and heat-tolerant wheat genotypes (Giza-168 and Giza-171) were followed by Sids-12 and Sids-14 wheat genotypes, which were able to maintain higher dry matter and pigment stability due to increased proline, lower accumulation of H2O2, MDA contents, and reduced oxidative stress with enhanced antioxidant capacity. The findings of the current study can be used in breeding programs to generate new promising wheat genotypes with improved drought and heat tolerance/resistance. In addition, our results provide a foundation for future molecular research in wheat at the seedling stage, modifying the antioxidant system’s related biochemical responses to better adapt wheat to early-stage stress. More research is needed to construct a pathway for converting data from the seedling stage to the other developmental phases, including yield and related characteristics. Several bioactive and biosafety features have been identified in SeNPs. To use SeNPs commercially, it is necessary to identify any negative effects they may have on agriculture and ecosystems.Drought and heat are the two significant environmental variables limiting agricultural yield worldwide. The primary scientific issues are improving drought and heat tolerance and eliminating the detrimental effects of stress in wheat. The current study concluded that foliar treatment with 10 mg·L


The physical vapor deposition (PVD) technique was successfully used to deposit the mono nanocomposite thin film with good adhesion and film thickness ≅ 100 ± 3 nm. The structural and morphological properties of the [PoPDA/TiO2]MNC thin films were studied by X-ray diffraction (XRD) and scanning electron microscope (SEM). The measured optical properties of the [PoPDA/TiO2]MNC thin films such as reflectance (R) in the UV–Vis-NIR spectrum, absorbance (Abs), and transmittance (T) were employed to probe the optical characteristics at room temperatures. As well as the calculations of TD-DFT , optimization through the TD-DFTD/Mol3 and Cambridge Serial Total Energy Bundle (TD-DFT/CASTEP) was employed to study the geometrical characteristics. The dispersion of the refractive index was examined by the single oscillator Wemple–DiDomenico (WD) model. Moreover, the single oscillator energy  was synthesized via the oxidative polymerization route. A poly(o-phenylene diamine) (PoPDA)/titanium dioxide nanoparticle mono nanocomposite [PoPDA/TiO One of the most crucial approaches to meeting the rising global energy needs using a renewable resource is to directly harvest energy from sunlight using photovoltaic technology. Due to the possibility of fabricating polymeric solar cells (PSCs) onto large regions of lightweight flexible substrates by solution processing at a cheap cost, they represent a promising alternative for creating clean and renewable energy ,2. Organ3 and CASTEP methodologies) in investigating the structure and phase stability [2]MNC nanostructured thin films for inserting into the devices such as solar cells. A significant role is also the control of the defects and impurity states in the organic molecular crystals to achieve proper photoelectrical characteristics. Moreover, [PoPDA/TiO2]MNC is structurally characterized by XRD. The thin film of [PoPDA/TiO2]MNC is fabricated by deposition on top of glass using (PVD) with annealing at 298 K for 1 h, and then, its optical characteristics investigated. In addition, the experimental data are investigated by dispersion models for the real refractive index and extinction coefficient. Moreover, the role of doping on optical properties is demonstrated.In the last 10 years, many reports have exhibited the crucial role of the contemporary TD-DFT applications .None of the compounds were further purified before usage. Ortho phenylenediamine was obtained from Across Organics, Thermo Fisher Scientific. The following substances were used as purchased: ferric chloride (Aldrich), ethanol, anhydrous dimethyl formaldehyde (DMF), dimethyl sulfoxide (DMSO), and polyethylene glycol (PEG200) from Shanghai Chemicals . Sigma-Aldrich provided us with hydrochloric, hydrofluoric, nitric, and acetic acids as well as a single crystal of p-Si and tetrahydrofuran (THF) and titanium dioxide . Then, 4 mL of PEG200 was added to the oPDA solution (as a soft surfactant to control the shape of the resultant polymer). After that, 6–7 mL of concentrated HCl was added to the oPDA solution while it was being stirred with a magnetic stirrer at 650 rpm and 25 °C. Under the previously mentioned circumstances, 160 mL (0.5 M) anhydrous ferric chloride solution was then gradually added to the oPDA solution as an oxidizing agent. It was discovered that the resulting solution had a pH of 0. The monomer and initiator are at a 1:1 ratio. The PoPDA that was produced was for seven days at room temperature. To remove of the extra oxidizing agent, surfactant, and protonated monomer, the resultant PoPDA was repeatedly rinsed with distilled water. At 60 °C, the resultant PoPDA was dried.2]MNC solution was doped with x = 3.0, 5.0, and 10.0 wt% of the sol–gel-prepared [TiO2]NPs. The required mass . Thin films were deposited onto a single crystal of (p-Si) wafer and an ITO/glass substrate using a UNIVEX 250 Leybold (Germany) at a base pressure of 5 × 103 Pa, interdigitated electrodes spaced by 75 m, and a deposition rate of 3 2]MNC were balanced using a quartz crystal microbalance on a UNIVEX 250 Leybold machine  was dissolved in 80 mL tetrahydrofuran (THF) under stirring at 65 °C for 35 min in tightly closed beakers under magnetic stirrer for additional 35 min at room temperature. To prepare nanocomposite films, the [PoPDA/TiOIso and nanocomposite [PoPDA/TiO2]Iso in the TD-DFT in gas phases state estimated evidence of PBE/GGA functionality, natural pseudo-positive preservatives, and a straightforward DNP set for acceptable compounds, DMol3 was used [Iso and nanocomposite [PoPDA/TiO2]Iso in gaseous state were detected using DMol3 IR features, leading to a GP frequency approximation. Three more factors have been found to alter how Becke functions [2]Iso, and the nanofluid in the gaseous phase have all been improved with WBX97XD/6-311G. The symmetric variables, images of enhancements, power, and vibration of processed nanocomposite mixtures are examined by the GAUSSIAN 09W system programming. Numerous useful details about the relationship between the setup and the variety of results offered by our group have been revealed by the WBX97XD/6-311G B3LYP technique [Iso and nanocomposite [PoPDA/TiO2]Iso Gaussian in isolated molecules, deliberate variations in descriptors, prototype vitality data, and the employment of various modifications with varying difficulties were evaluated using the GAUSSIAN 09W and DMOl3 methodologies.In order to examine the frequency studies and molecular structure performance for the polymer [PoPDA]was used ,51. The unctions . B3LYP Lunctions  Form andechnique . The porTF and nanocomposite [PoPDA/TiO2]MNC were recorded by utilizing the following instruments: the powder X-ray diffraction (XRD) analysis at 2θ = 4–90°, its collected data for the line broadening on the synthesized powder and phase composition by using a Shimadzu apparatus with Xpert diffractometer . The surface morphology of the produced [PoPDA]TF and [PoPDA/TiO2]MNC thin films were studied utilizing a scanning electron microscope (SEM) (QUANTA FEG 250) and EDX unite. The TEM images were captured at an accelerating voltage of 120 kV utilizing a JEM 200CX transmission electron microscope . The optical properties and absorbance of the organic compounds were computed on a PerkinElmer Lambda 365 spectrophotometer. Photoluminescence was measured with a Hitachi F-7100 fluorescence spectrometer equipped with a detector photomultiplier R928F. Current density versus voltage (J–V) curves of the [PoPDA/TiO2]MNC were measured by a Keithley 2400 source meter in air conditions that scanned from 300 nm to 700 nm, under the 100 ± 3 nm illuminations with intensities of 300 mW/cm2, 100 mW/cm2, and 50 mW/cm2, respectively. The monochromatic light used in all these measurements was provided by a 150 W xenon lamp coupled with a monochromator. EQE is calculated as:R is the responsivity, The various methods of analysis and typical settings that describe the polymer [PoPDA]TF and [PoPDA/TiO2]MNC thin film are shown in TF and [PoPDA/TiO2]MNC thin films have strong diffraction peaks in the range 10° ≤ 2θ ≤ 60° and 20° ≤ 2θ ≤ 60°, respectively, which indicate the long-range and crystallinity order. The estimated crystalline structure of [PoPDA]TF and [PoPDA/TiO2]MNC thin films, crystal device variance, hkl, d-spacing (d), and FWHM (β) are shown in Iso thin film with the parameters a = 13.95(6) Ǻ, b = 13.83(3) Ǻ, c = 9.943(9) Ǻ, α = 90°, β = 90°, γ = 90°, and volume = 1910(8) Ǻ3 [2]Iso thin film with a MONOCLINIC space group unit cell with the parameters a = 13.342(1) Ǻ, b = 13.127(2) Ǻ, c = 8.994(2) Ǻ, α = 90°, β = 100.28°, γ = 90°, and volume = 1550.16(2) Ǻ3. The [PoPDA]TF and [PoPDA/TiO2]MNC thin films’ average crystalline sizes (DAv) were found to be 102.5 and 88.7 nm, respectively (β) and crystalline size D were measured in the range 10°≤ 2θ ≤ 60° for [PoPDA]Iso and listed in λ = 0.154 nm and β is in radians. The database code amcsd 0001206 is in excellent agreement with the interplanar distance d-spacing. Peak lines estimated by the TD-DFT-DFT and Crystal Sleuth Microsoft applications are close to the measured data [X-ray diffraction pattern (XRD) studies were applied to determine the crystalline phase configurations and atomic orientation of thin film and multi-layered films. Experimental XRD studies for [PoPDA]10(8) Ǻ3 . Figure ectively . It is kred data .2] as isolated molecules were investigated using electron density and electrostatic potential as shown in The chemical and physical features of the gaseous phase of the polymer [PoPDA] and nanocomposite [PoPDA/TiOIso and [PoPDA/TiO2]Iso , which was calculated based on surface density, increased the likelihood of electron transfer [PoPDA]Iso and [PoPDA/TiO2]Iso in the gaseous phase.In TD-DFT and TD-DFT/Gaussian ideas, the electron density was used as a crucial operator to explore the isolated electron system for [PoPDA]TiO2]Iso a. FigureIso and [PoPDA/TiO2]Iso, based on the data in −1 ≤ [MEP] ≤ 8.608 × 10−2 and −2.072 × 10−1 ≤ [MEP]≤ 2.491 × 10−1 for [PoPDA]Iso and [PoPDA/TiO2]Iso, respectively, in the isolated phase of the molecule. The MEP diagram also revealed the potential negative regions of the hydrogen atoms’ positive potential. Red, brown, and blue were listed as the order of the colors [2=CH–) was represented in [PoPDA]Iso and [PoPDA/TiO2]Iso, with a maximum value of +3.87 a.u. According to the MEP map computation, the sites with positive potential were (–CH2C–), while the sites with negative potential were (–C=N), (CH2=CH–), and (–CH2C–). Molecule intermolecular interaction has given remarkably evidence for these locations. Consequently, intermolecular hydrogen-bonding existence is emphasized in e colors . Blue ine colors ,63 Addit3TD-DFT/DMOl procedure, LUMO  and HOMO  states as presented in LUMO and HOMO states impact the fragment molecular orbital (FMO) method’s complicated analysis in quantum chemical simulations. The computed energy values of μ), global hardness (S), the global index of electrophilicity (ω), the electronic charge maximum amount (Iso and [PoPDA/TiO2]IsoConsidering the advantage of the vity (χ) ,65 of MNC thin films are shown in TF in 2]MNC surface is shown in 2]MNC film, as well as the rates of relative evaporation and condensation, in which the volume and size of the pore were determined [SEM images of the [PoPDA]termined .Generally, the variety of nanoparticle shapes that can be used is an important detail in the composition of hybrid solar cells. A specific shape of nanoparticles may improve the efficiency and performance of hybrid solar cells, but no direct relationship has been demonstrated. Lazaro A. et al. , for exaTF and [PoPDA/TiO2]MNC  [2]MNC show a disappearance of the absorption peak at 387 nm and a new absorption peak appears at 480 nm, which belongs to TiO2 nanoparticles. The addition of TiO2 nanoparticles results in a reduction in the 332 nm absorption peak intensity to 33.33% which agrees with the previous work MNC are 2.296 eV and 2.114 eV, respectively, in the case of direct transition, and 3.511 eV and 3.209 eV, respectively, in the case of indirect transition. The calculated values of the direct and indirect energy gaps show a significant decrease in the direct and indirect energy gap values by adding TiO2 nanoparticles and indicate the preference for direct electron transition.Using absorption coefficient α, which was calculated using the relation: Interaction between incident light and polymer nanocomposites plays an important role in using polymer nanocomposites in different applications such as optical devices and solar cells. The refractive index n(λ) and absorption index k(λ) indicate how much incident light is refracted and absorbed when interacting with the thin film. The absorption index k(λ) and refractive index n(λ) were calculated using the equations:TF and decreased to ranges between 1.63 and 1.29 for [PoPDA/TiO2]MNC, which indicates the improvements in the optical properties.On the other hand, the similarity in the behavior between the absorption index k(λ) and absorption spectra in 1 and ε2 represent the material’s ability to store electric energy and the ohmic resistance of the material. Depending on the refractive index n(λ) and absorption index k(λ) values, the real and imaginary parts of the dielectric can be calculated asReal and imaginary parts of the dielectric constant ε1(ω) and ε2(ω) as a function of photon energy, and similar behavior of the simulated ε1(ω) and ε2(ω) is observed  reaches its maximum intensity of 2.961 (F·m−1) at hν = 3.725 eV, while ε2(ω)’s maximum intensity was 4.49 × 10−8 (F·m−1) at the same photon energy. On the other hand, ε1(ω) and ε2(ω) reached their maximum intensities of 2.667 and 4.309 × 10−8 (F·m−1), respectively, at hν = 2.569 eV for [PoPDA/TiO2]MNC. High values of ε1(ω) for both [PoPDA]TF and [PoPDA/TiO2]MNC indicate a great capacity to store electric energy, while low values of ε2(ω) indicate a good electrical conductivity behavior. DFT-CASTEP simulations of dielectric constants show a good agreement with dielectric experimental data.observed b. For TF, while for [PoPDA/TiO2]MNC, σ2(ω) has a different behavior than σ1(ω), which is a result of adding TiO2 nanoparticles. Simulated data using DFT-CASTEP simulations in TF and [PoPDA/TiO2]MNC in d and single oscillator energy Eo (related to direct energy gap by the relation Eo ≈ 1.5 → 2 Eg) using the following equation [The Wemple–DiDomenico model is applied on [PoPDA]equation :(19)nlculated c. n∞, λoTF and nanocomposite [PoPDA/TiO2]MNC are represented in The comparative studies for the nanoblend [PoPDA]TF and [PoPDA/TiO2]MNC thin films in the range of 1.5 to 6.25 eV are shown in TF, the main emission peak appeared at 2.469 eV; this peak is related to the excess of fluorophore molecules in the polymer chain. In the case of [PoPDA/TiO2]MNC, the emission peak, in blue, shifted to 3.297 eV; this shift is due to the addition of TiO2 nanoparticles. A shift of 0.828 eV occurred when TiO2 nanoparticles were added to the PoPDA polymer, which is a fluorescent polymer due to the random lasing in the weakly scattering system. In random lasing cases, the feedback does not occur in conventional resonant cavities, but the introduction of nano-scatterers (TiO2 nanoparticles in this case) as a disordering factor strongly influences the feedback mechanism [Photoluminescence spectra for [PoPDA]echanism .TF and [PoPDA/TiO2]MNC, for [PoPDA]TF digital photographs obtained at hν = 2.469 eV, which is located at the coordinates , while the digital photographs for [PoPDA/TiO2]MNC were obtained at hν = 3.297 eV and located at the coordinates . A significant change in the color of the emission can be figured out since the emission color changed from light purple for [PoPDA]TF to pale blue for [PoPDA/TiO2]MNC. The effect of adding TiO2 to PoPDA to fabricate [PoPDA/TiO2]MNC results in making it close to emitting white light from a single material, giving it potential applications in lighting and display devices.Commission Internationale de l’Eclairage (CIE) graphs  illustra2]MNC/n-Si/Al heterojunction diode are shown in 2). As shown in oc changed from 0 V in the dark condition to 0.135 V in the illuminated condition; at a bias of +2 V, the device at different illumination intensities of 0, 50, 100, and 200 mW/cm2 showed forward currents of 0.001, 0.166, 0.667, and 1.828 mA/cm2, respectively. It is known that the quality of the diode is determined by the ratio of forwarding current to reverse current (JF/JR); the forward currents of the Au/[ PoPDA/TiO2]MNC/n-Si/Al heterojunction diode at +2 V are 0.001, 0.166, 0.667, and 1.828 mA/cm2, respectively, while the reverse currents at V = −2 V are 0.0044, 0.46, 1.4, and 5.8 μA/cm2, respectively, for the illuminations 0, 50, 100, and 200 mW/cm2, respectively. The resulting rectification ratios (JF/JR) are 0.22, 0.36, 0.47, and 0.31 × 103, respectively, at ±2 V.Dark and illuminated J-V characteristic curves for the Au/TF/n-Si/Al and Au/[ PoPDA/TiO2]MNC/n-Si/Al heterojunction diodes. Moreover, the values of r2 take the same behavior as r1 with temperature, the values of r2 decrease by raising T (K), and the polymers’ conduction mechanism is stated by the nonlinear coefficient parameters (r). When r = 1 we can obtain the ohmic behavior, and r = 2 (a dominant mechanism) indicates the incomplete trap-free space charge. Finally, the case when r > 2 indicates the defectiveness of the mechanism in terms of the trap charge.The nonlinear coefficient parameter r can be deduced using the relation: 2 nanoparticles interact with the polymer by a weak interfacial contact and TiO2 nanoparticles aggregate as a result of van der Waals force, uniform dispersion is then difficult to achieve. In this study, TiO2 nanoparticles dispersed excellently in the polymer matrix as indicated by the SEM and AFM images. 2 nanoparticles interacted with PoPDA in the nanocomposite film. The σdc of the [PoPDA/TiO2]MNC thin film was 66.06 × 10−5 S m−1, which is much larger than the value of 10.09 × 10−5 S m−1 for the [PoPDA]TF thin film. The values of Ln σdc ranged between (−7.49 and−8.79) at 1/T = 0.0026 1/K and (−3.18 and −3.95) at 1/T = 0.002359 1/K. There were two problems that occurred in order to increase TiO2 nanoparticles’ loading: (1) the viscosity of PoPDA, which makes it difficult to absorb additional TiO2 nanoparticles and (2) the high surface energy of TiO2 nanoparticles, which makes them tend to agglomerate. An appropriate applied electrical field (E) tends to align TiO2 nanoparticles, which results in an agglomeration decrease and the network expands from the negative to the positive electrode. Conducting pathways are generated by following the stages: (1) the rotating of TiO2 nanoparticles to a specific angle according to the applied E, resulting in dipole moment generation at TiO2 nanoparticles edges, which makes them align to the electric field direction, (2) a 3D network is created by the attraction between the TiO2 nanoparticles until they make contact, and (3) the moving and adherence of TiO2 nanoparticles to the negative electrode. In conclusion, by ignoring the ionic conductivity, σdc of the nanocomposite films is primarily caused by the electronic conductivity of the TiO2 nanoparticles. The charge transfer is increased by increasing the temperature T (K), as seen in TF and the low concentrations of TiO2 nanoparticles, there were no 3D networks, but they would be activated by the charge carrier collected energy to leap potential barriers. Finally, heating and increasing the TiO2 nanoparticles’ content will optimize these routes, increase σdc, and develop networks MNC/p-Si/Al polymer solar cell. I-V curves of the solar cell were measured by a computerized Keithley 2635A system. A GPIB/USB cable was used to connect the source meter to the host computer. The photovoltaic performance, saturation current density (JSC), and open-circuit voltage (VOC) of the manufactured solar cell are listed in SC) and (VOC) were increased by increasing the incident light intensity. A prominent photovoltaic response was exhibited from the Au/[ PoPDA/TiO2]MNC/p-Si/Al heterojunction device since many high rectification degrees were exhibited in J-V characteristics (sc is related to the incident light intensity by the relation:mJ), voltage density (mV), and maximum power (mP) are tabulated in in). It can be figured out that mJ, mV, and mP are increased by increasing the light intensity. The fill factor (FF) and power conversion efficiency of the Au/[ PoPDA/TiO2]MNC/p-Si/Al heterojunction device were calculated using the equations:FF and 2]MNC/p-Si/Al heterojunction device is improved by increasing the light intensity. At Pin = 200 mW/cm2, FF and Excited state photo-physics generated from absorption in the aggregate state can be investigated easily from the photovoltaic response. Moreover, increasing the nanoparticle content  and nanocomposite Au/[PoPDA/TiO2]MNC/p-Si/Al (B), more electrons can be easily injected from P-Si onto the LUMO of the nanoblend and nanocomposite due to the small injection barrier of 0.4 eV and the large contact interface between P-Si and the nanoblend and nanocomposite. The injected electrons can be efficiently transported in the active layer along the continuous electron transport channels due to the nanoblend and nanocomposite, resulting in the relatively high dark current and high withstanding reverse bias up to 19 V. In order to investigate the photo response of all the polymer photodetectors (PPDs) with the nanoblend and composite, EQE spectra of all PPDs were measured under the given reverse biases and are shown in 2]NPs under reverse bias. An interesting phenomenon is that there is a distinct dip from about 490 nm to 570 nm and two apparent peaks at about 380 nm and 625 nm in the EQE spectra of a trap-assisted photomultiplication (PM)-phenomenon-type PPD. It is very apparent that the EQE spectra of the PM-type PPDs cannot match the absorption spectrum of the nanoblend or nanocomposite.For the devices, nanoblend Au/[PoPDA]2]NPs and poly orthophenylene diamine in powder form. Physical vapor deposition (PVD) technology was used to create thin films of [PoPDA]TF polymer and [PoPDA/TiO2]MNC composite measuring 100 ± 3 nm in thickness at room temperature. Using a quartz crystal microbalance, the [PoPDA/TiO2]MNC thin film’s thickness was measured using a UNIVEX 250 Leybold machine. The structural properties were investigated using XRD, SEM, and AFM. According to XRD calculations, the typical crystallite sizes of [PoPDA]TF and [PoPDA/TiO2]MNC are 102.5 nm and 88.7 nm, respectively. The TD-DFT calculations accurately matched the observed XRD and optical spectra and validated the molecular structure of the examined materials. The values of the optical energy gap TF and [PoPDA/TiO2]MNC are 2.296 eV and 2.114 eV, respectively, in the case of direct transition, and 3.511 eV and 3.209 eV, respectively, in the case of indirect transition. The bandgap energy of the pristine polymer can be reduced by adding [TiO2]NPs to the polymer, reducing the bandgap energy by a factor of 51.29%. As established by TD-DFT/DMol3, the isolated molecule of the composite [PoPDA/TiO2]Iso has a band gap of 2.558 eV. By calculating the values of 2]NPs nanoparticles, the DC conductivity characteristics of the polymer nanocomposite can be improved by adding these particles. The Au/[PoPDA/TiO2]MNC/n-Si/Al polymer solar cell can be made using [PoPDA/TiO2]MNC. The Au/[ PoPDA/TiO2]MNC/n-Si/Al heterojunction device’s fill factor (FF) and power conversion efficiency are 68.04% and 19.69 at 200 W·cm−2, respectively.In this study, oxidative polymerization was used to synthesize a polycrystal-solid form of the poly orthophenylene diamine polymer. Additionally, the sol–gel process was used to create the novel composite made of titanium dioxide nanoparticles [TiO