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Investigation of the Influence of Polyamines on Mature Embryo Culture and DNA Methylation of Wheat (Triticum aestivum L.) Using the Machine Learning Algorithm Method
Numerous factors can impact the efficiency of callus formation and in vitro regeneration in wheat cultures through the introduction of exogenous polyamines (PAs). The present study aimed to investigate in vitro plant regeneration and DNA methylation patterns utilizing the inter-primer binding site (iPBS) retrotransposon and coupled restriction enzyme digestion–iPBS (CRED–iPBS) methods in wheat. This investigation involved the application of distinct types of PAs (Put: putrescine, Spd: spermidine, and Spm: spermine) at varying concentrations (0, 0.5, 1, and 1.5 mM). The subsequent outcomes were subjected to predictive modeling using diverse machine learning (ML) algorithms. Based on the specific polyamine type and concentration utilized, the results indicated that 1 mM Put and Spd were the most favorable PAs for supporting endosperm-associated mature embryos. Employing an epigenetic approach, Put at concentrations of 0.5 and 1.5 mM exhibited the highest levels of genomic template stability (GTS) (73.9%). Elevated Spd levels correlated with DNA hypermethylation while reduced Spm levels were linked to DNA hypomethylation. The in vitro and epigenetic characteristics were predicted using ML techniques such as the support vector machine (SVM), extreme gradient boosting (XGBoost), and random forest (RF) models. These models were employed to establish relationships between input variables (PAs, concentration, GTS rates, Msp I polymorphism, and Hpa II polymorphism) and output parameters (in vitro measurements). This comparative analysis aimed to evaluate the performance of the models and interpret the generated data. The outcomes demonstrated that the XGBoost method exhibited the highest performance scores for callus induction (CI%), regeneration efficiency (RE), and the number of plantlets (NP), with R2 scores explaining 38.3%, 73.8%, and 85.3% of the variances, respectively. Additionally, the RF algorithm explained 41.5% of the total variance and showcased superior efficacy in terms of embryogenic callus induction (ECI%). Furthermore, the SVM model, which provided the most robust statistics for responding embryogenic calluses (RECs%), yielded an R2 value of 84.1%, signifying its ability to account for a substantial portion of the total variance present in the data. In summary, this study exemplifies the application of diverse ML models to the cultivation of mature wheat embryos in the presence of various exogenous PAs and concentrations. Additionally, it explores the impact of polymorphic variations in the CRED–iPBS profile and DNA methylation on epigenetic changes, thereby contributing to a comprehensive understanding of these regulatory mechanisms.
Genotype-Trait (GT) Biplot Analysis for Yield and Quality Stability in Some Sweet Corn (Zea mays L. saccharata Sturt.) Genotypes
A strong statistical method for investigating the correlations between traits, assessing genotypes based on numerous traits, and finding individuals who excel in particular traits is genotype–trait (GT) biplot analysis. The current study was applied to evaluate 11 sweet corn (Zea mays L. saccharata) genotypes and correlate them based on genotype–trait (GT) biplot analysis for two cropping seasons in Erzurum, Türkiye using the RCBD experimental design with three reputations. The results showed that the genotypes were significantly different for the majority of the examined variables according to the combined analysis of variance findings at 0.01 probability level. An ecological analysis was performed to evaluate sweet corn varieties and environmental conditions and interactions between them (genotype × environmental conditions). Our results showed that the summation of the first two and second main components was responsible for 73.51% of the combined cropping years of the sweet corn growth and development variance, demonstrating the biplot graph’s optimum relative validity, which was obtained. In this study, the Khan F1 (G6) genotype was found to be the stablest genotype, and the Kompozit Seker (G7) genotype was the non-stable genotype, moreover based on the first cropping year, second cropping year, and the average mean of the two cropping years. As a conclusion, the Khan F1 (G6) genotype is the highest-yielding genotype, and the Kompozit Seker (G7) is the lowest. Based on the heat map dendrogram, the context of the differential extent of trait association of all genotypes into two clusters is indicated. The highest genetic distance was shown between the BATEM Tatlı (G3) and Febris (G5) genotypes. Our results provide helpful information about the sweet corn genotypes and environments for future breeding programs.
Ethyl Methanesulfonate (EMS) Mutagen Toxicity-Induced DNA Damage, Cytosine Methylation Alteration, and iPBS-Retrotransposon Polymorphisms in Wheat (Triticum aestivum L.)
The use of mutagens in plant breeding is used to create new germplasm, increase agricultural yield, quality, and resistance to diseases and pests. Mutagens are physical or chemical factors that can alter the DNA or RNA structure of an organism, causing mutations above the expected level. One of the most common and potent chemical mutagens is EMS (ethyl-methane sulfonate), which produces point mutations in plants, but to a lesser degree can also cause the loss or deletion of a chromosomal region. This study used inter-primer binding site (iPBS) and coupled restriction enzyme digestion inter-primer binding site (CRED-iPBS) technique analysis to determine the effect of EMS mutagens on methylation rates in wheat genotypes at seedling growth stage. Treatments with five different EMS concentrations (0%; control, 0.1%, 0.2%, 0.3%, and 0.4%) at four different times (0; control, 3, 6, and 9 h) were used. Inter-primer binding site (iPBS) markers were employed to assess genomic instability and cytosine methylation in treated wheat. In seeds treated with EMS at different concentrations and times, the disappearance of regular bands and the formation of new bands due to the effects of the EMS mutagen revealed that genetic diversity exists. The CRED-iPBS analysis revealed that the 3 h + 0.1% EMS treatment produced the highest MspI polymorphism value (19.60%), while the 9 h + 0.1% EMS treatment produced the lowest value (10.90%). The mutagenic effects of EMS treatments had considerable polymorphism on a variety of impacts on the cytosine methylation and genomic instability of wheat. According to the current research, EMS mutagenesis may be a practical method for accelerating breeding programs to produce enough genetic diversity in wheat populations. Mutation-assisted breeding and the subsequent selection of desirable mutants using genetic markers may also be carried out in wheat utilizing an integrated strategy.
Identification of Novel QTLs Associated with Frost Tolerance in Winter Wheat (Triticum aestivum L.)
Low temperature (cold) and freezing stress is a major problem during winter wheat growth. Low temperature tolerance (LT) is an important agronomic trait in winter wheat and determines the plants’ ability to cope with below-freezing temperatures; thus, the development of cold-tolerant cultivars has become a major goal of breeding in various regions of the world. In this study, we sought to identify quantitative trait loci (QTL) using molecular markers related to freezing tolerance in winter. Thirty-four polymorphic markers among 425 SSR markers were obtained for the population, including 180 inbred lines of F12 generation wheat, derived from crosses (Norstar × Zagros) after testing with parents. LT50 is used as an effective selection criterion for identifying frost-tolerance genotypes. The progeny of individual F12 plants were used to evaluate LT50. Several QTLs related to wheat yield, including heading time period, 1000-seed weight, and number of surviving plants after overwintering, were identified. Single-marker analysis illustrated that four SSR markers with a total of 25% phenotypic variance determination were linked to LT50. Related QTLs were located on chromosomes 4A, 2B, and 3B. Common QTLs identified in two cropping seasons based on agronomical traits were two QTLs for heading time period, one QTL for 1000-seed weight, and six QTLs for number of surviving plants after overwintering. The four markers identified linked to LT50 significantly affected both LT50 and yield-related traits simultaneously. This is the first report to identify a major-effect QTL related to frost tolerance on chromosome 4A by the marker XGWM160. It is possible that some QTLs are closely related to pleiotropic effects that control two or more traits simultaneously, and this feature can be used as a factor to select frost-resistant lines in plant breeding programs.
Sodium Azide as a Chemical Mutagen in Wheat (Triticum aestivum L.): Patterns of the Genetic and Epigenetic Effects with iPBS and CRED-iPBS Techniques
Wheat, which is scientifically known as Triticum aestivum L., is a very nutritious grain that serves as a key component of the human diet. The use of mutation breeding as a tool for crop improvement is a reasonably rapid procedure, and it generates a variety that may be used in selective breeding programs as well as functional gene investigations. The present experiment was used to evaluate the potential application of a conventional chemical mutagenesis technique via sodium azide (NaN3) for the germination and seedling growth stage in wheat. Experiments with NaN3 mutagenesis were conducted using four different treatment periods (0, 1, 2, and 3 h) and five different concentrations (0, 0.5, 1, 1.5, and 2 mM). The genomic instability and cytosine methylation of wheat using its seeds were investigated after they were treated. In order to evaluate the genomic instability and cytosine methylation in wheat that had been treated, interprimer binding site (iPBS) markers were used. The mutagenic effects of NaN3 treatments had considerable polymorphism on a variety of impacts on the cytosine methylation and genomic instability of wheat plants. The results of the experiment showed considerable changes in the iPBS profiles produced by the administration of the same treatments at different dosages and at different times. Coupled restriction enzyme digestion interprimer binding site (CRED-iPBS) assays identified changes in gDNA cytosine methylation. The highest polymorphism value was obtained during 1 h + 2 mM NaN3, while the lowest (20.7%) was obtained during 1 h + 1.5 mM NaN3. Results showed that treatments with NaN3 had an effect on the level of cytosine methylation and the stability of the genomic template in wheat plants in the germination stage. Additionally, an integrated method can be used to for mutation-assisted breeding using a molecular marker system in wheat followed by the selection of desired mutants.
Prediction of Grain Yield in Wheat by CHAID and MARS Algorithms Analyses
Genetic information obtained from ancestral species of wheat and other registered wheat has brought about critical research, especially in wheat breeding, and shown great potential for the development of advanced breeding techniques. The purpose of this study was to determine correlations between some morphological traits of various wheat (Triticum spp.) species and to demonstrate the application of MARS and CHAID algorithms to wheat-derived data sets. Relationships among several morphological traits of wheat were investigated using a total of 26 different wheat genotypes. MARS and CHAID data mining methods were compared for grain yield prediction from different traits using cross-validation. In addition, an optimal CHAID tree structure with minimum RMSE was obtained and cross-validated with nine terminal nodes. Based on the smallest RMSE of the cross-validation, the eight-element MARS model was found to be the best model for grain yield prediction. The MARS algorithm proved superior to CHAID in grain yield prediction and accounted for 95.7% of the variation in grain yield among wheats. CHAID and MARS analyses on wheat grain yield were performed for the first time in this research. In this context, we showed how MARS and CHAID algorithms can help wheat breeders describe complex interaction effects more precisely. With the data mining methodology demonstrated in this study, breeders can predict which wheat traits are beneficial for increasing grain yield. The adaption of MARS and CHAID algorithms should benefit breeding research.
Zinc Oxide Nanoparticles: An Influential Element in Alleviating Salt Stress in Quinoa (Chenopodium quinoa L. Cv Atlas)
Climate change has intensified abiotic stresses, notably salinity, detrimentally affecting crop yield. To counter these effects, nanomaterials have emerged as a promising tool to mitigate the adverse impacts on plant growth and development. Specifically, zinc oxide nanoparticles (ZnO-NPs) have demonstrated efficacy in facilitating a gradual release of zinc, thus enhancing its bioavailability to plants. With the goal of ensuring sustainable plant production, our aim was to examine how green-synthesized ZnO-NPs influence the seedling growth of quinoa (Chenopodium quinoa L. Cv Atlas) under conditions of salinity stress. To induce salt stress, solutions with three different NaCl concentrations (0, 100, and 200 mM) were prepared. Additionally, Zn and ZnO-NPs were administered at four different concentrations (0, 50, 100, and 200 ppm). In this study, plant height (cm), plant weight (g), plant diameter (mm), chlorophyll content (SPAD), K/Na value, Ca/Na value, antioxidant enzyme activities (SOD: EU g−1 leaf; CAT: EU g−1 leaf; POD: EU g−1 leaf), H2O2 (mmol kg−1), MDA (nmol g−1 DW), proline (µg g−1 FW), and sucrose (g L−1), content parameters were measured. XRD analysis confirmed the crystalline structure of ZnO nanoparticles with identified planes. Salinity stress significantly reduced plant metrics and altered ion ratios, while increasing oxidative stress indicators and osmolytes. Conversely, Zn and ZnO-NPs mitigated these effects, reducing oxidative damage and enhancing enzyme activities. This supports Zn’s role in limiting salinity uptake and improving physiological responses in quinoa seedlings, suggesting a promising strategy for enhancing crop resilience. Overall, this study underscores nanomaterials’ potential in sustainable agriculture and stress management.
Insights into Drought Tolerance of Tetraploid Wheat Genotypes in the Germination Stage Using Machine Learning Algorithms
Throughout germination, which represents the initial and crucial phase of the wheat life cycle, the plant is notably susceptible to the adverse effects of drought. The identification and selection of genotypes exhibiting heightened drought tolerance stand as pivotal strategies aimed at mitigating these effects. For the stated objective, this study sought to evaluate the responses of distinct wheat genotypes to diverse levels of drought stress encountered during the germination stage. The induction of drought stress was achieved using polyethylene glycol at varying concentrations, and the assessment was conducted through the application of multivariate analysis and machine learning algorithms. Statistical significance (p < 0.01) was observed in the differences among genotypes, stress levels, and their interaction. The ranking of genotypes based on tolerance indicators was evident through a principal component analysis and biplot graphs utilizing germination traits and stress tolerance indices. The drought responses of wheat genotypes were modeled using germination data. Predictions were then generated using four distinct machine learning techniques. An evaluation based on R-square, mean square error, and mean absolute deviation metrics indicated the superior performance of the elastic-net model in estimating germination speed, germination power, and water absorption capacity. Additionally, in assessing the criterion metrics, it was determined that the Gaussian processes classifier exhibited a better performance in estimating root length, while the extreme gradient boosting model demonstrated superior performance in estimating shoot length, fresh weight, and dry weight. The study’s findings underscore that drought tolerance, susceptibility levels, and parameter estimation for durum wheat and similar plants can be reliably and efficiently determined through the applied methods and analyses, offering a fast and cost-effective approach.
Genetic Diversity and Population Structure in Bread Wheat Germplasm from Türkiye Using iPBS-Retrotransposons-Based Markers
This study investigated the genetic diversity and population structure of 63 genotypes from Turkish bread wheat germplasm using iPBS-retrotransposons primers. The thirty-four iPBS primers produced a total of 1231 polymorphic bands, ranging from 8 (iPBS-2375) to 60 (iPBS-2381) alleles per marker, with an average number of 36.00 alleles. The polymorphism information content (PIC) per marker varied between 0.048 (iPBS 2087) and 0.303 (iPBS 2382), with an average of 0.175. The numbers of effective alleles (ne), genetic diversity of Nei (h), and Shannon’s information index (I) value were calculated as 1.157, 0.95, and 0.144, respectively. The greatest genetic distance (0.164) was between Eastern Anatolia Agricultural Research Institute genotypes and Çukurova Agricultural Research Institute genotypes. The unweighted pair-group method with arithmetic mean (UPGMA) dendrogram placed the 63 wheat genotypes into three clusters. The percentage of genetic diversity explained by each of the three main coordinates of the basic coordinate analysis was determined to be 44.58, 12.08, and 3.44, respectively. AMOVA (Analysis of Molecular Variance) showed that the variation within populations was 99% and that between populations was 1%. The result of genetic structure analysis suggests that the greatest value of K was calculated as 3. The F-statistic (Fst) value was determined as 0.4005, 0.2374, and 0.3773 in the first to third subpopulations, respectively. Likewise, the expected heterozygosity values (He) were determined as 0.2203, 0.2599, and 0.2155 in the first, second, and third subpopulations, respectively. According to the information obtained in the study, the most genetically distant genotypes were the G1 (Aksel 2000) and G63 (Karasu 90) genotypes. This study provided a deep insight into genetic variations in Turkish bread wheat germplasm using the iPBS-retrotransposons marker system.