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Öğe Assessment of genetic diversity and population structure in local alfalfa genotypes using iPBS molecular markers(Springer, 2023) Eren, Baris; Keskin, Bilal; Demirel, Fatih; Demirel, Serap; Turkoglu, Aras; Yilmaz, Abdurrahim; Haliloglu, KamiliPBS retrotransposon markers, in other words, inter-primer binding site markers based on retrotransposon, have been helpful for the determination of genetic diversity in several plants. The study was evaluated using 10 iPBS molecular markers on the level of genetic diversity and genetic structure of alfalfa genotypes. A total of 280 alleles in 50 alfalfa genotypes (48 local genotypes and 2 commercial varieties) were obtained by 267 polymorphic markers with an average of 28 per locus, ranging from 9 to 41 alleles. The rate of polymorphism of the markers ranged from 86.36 to 100% with an average of 93.71%. The average number of polymorphic bands per marker was detected as 26.7. The mean PIC value and Dice's similarity index were calculated as 0.14 and 0.50 respectively. The results of UPGMA analysis, principal coordinate analysis (PCoA), and STRUCTURE demonstrated that the 50 alfalfa genotypes could be classified into 4 subpopulations, namely the Q1, Q2, Q3, and Q4. The Nei's genetic distances ranged from 0.0121 to 0.0359. iPBS markers and alfalfa genotypes used in this study may be used in studies of alfalfa breeding and germplasm conservation.Öğe Ethyl Methanesulfonate (EMS) Mutagen Toxicity-Induced DNA Damage, Cytosine Methylation Alteration, and iPBS-Retrotransposon Polymorphisms in Wheat (Triticum aestivum L.)(Mdpi, 2023) Turkoglu, Aras; Haliloglu, Kamil; Tosun, Metin; Bujak, Henryk; Eren, Baris; Demirel, Fatih; Szulc, PiotrThe 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.Öğe Investigation of the Influence of Polyamines on Mature Embryo Culture and DNA Methylation of Wheat (Triticum aestivum L.) Using the Machine Learning Algorithm Method(Mdpi, 2023) Eren, Baris; Turkoglu, Aras; Haliloglu, Kamil; Demirel, Fatih; Nowosad, Kamila; Ozkan, Guller; Niedbala, GniewkoNumerous 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 R-2 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 R-2 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.Öğe Machine Learning Analysis of the Impact of Silver Nitrate and Silver Nanoparticles on Wheat (Triticum aestivum L.): Callus Induction, Plant Regeneration, and DNA Methylation(Mdpi, 2023) Turkoglu, Aras; Haliloglu, Kamil; Demirel, Fatih; Aydin, Murat; Cicek, Semra; Yigider, Esma; Demirel, SerapThe objective of this study was to comprehend the efficiency of wheat regeneration, callus induction, and DNA methylation through the application of mathematical frameworks and artificial intelligence (AI)-based models. This research aimed to explore the impact of treatments with AgNO3 and Ag-NPs on various parameters. The study specifically concentrated on analyzing RAPD profiles and modeling regeneration parameters. The treatments and molecular findings served as input variables in the modeling process. It included the use of AgNO3 and Ag-NPs at different concentrations (0, 2, 4, 6, and 8 mg L-1). The in vitro and epigenetic characteristics were analyzed using several machine learning (ML) methods, including support vector machine (SVM), random forest (RF), extreme gradient boosting (XGBoost), k-nearest neighbor classifier (KNN), and Gaussian processes classifier (GP) methods. This study's results revealed that the highest values for callus induction (CI%) and embryogenic callus induction (EC%) occurred at a concentration of 2 mg L-1 of Ag-NPs. Additionally, the regeneration efficiency (RE) parameter reached its peak at a concentration of 8 mg L-1 of AgNO3. Taking an epigenetic approach, AgNO3 at a concentration of 2 mg L-1 demonstrated the highest levels of genomic template stability (GTS), at 79.3%. There was a positive correlation seen between increased levels of AgNO3 and DNA hypermethylation. Conversely, elevated levels of Ag-NPs were associated with DNA hypomethylation. The models were used to estimate the relationships between the input elements, including treatments, concentration, GTS rates, and Msp I and Hpa II polymorphism, and the in vitro output parameters. The findings suggested that the XGBoost model exhibited superior performance scores for callus induction (CI), as evidenced by an R-2 score of 51.5%, which explained the variances. Additionally, the RF model explained 71.9% of the total variance and showed superior efficacy in terms of EC%. Furthermore, the GP model, which provided the most robust statistics for RE, yielded an R-2 value of 52.5%, signifying its ability to account for a substantial portion of the total variance present in the data. This study exemplifies the application of various machine learning models in the cultivation of mature wheat embryos under the influence of treatments and concentrations involving AgNO3 and Ag-NPs.Öğe Mammalian Sex Hormones as Steroid-Structured Compounds in Wheat Seedling: Template of the Cytosine Methylation Alteration and Retrotransposon Polymorphisms with iPBS and CRED-iBPS Techniques(Mdpi, 2023) Demirel, Fatih; Turkoglu, Aras; Haliloglu, Kamil; Eren, Baris; Ozkan, Guller; Uysal, Pinar; Pour-Aboughadareh, AlirezaPhytohormones are chemical compounds found naturally in plants that have a significant effect on their growth and development. The increase in research on the occurrence of mammalian sex hormones (MSHs) in plants has prompted the need to investigate the functions performed by these hormones in plant biology. In the present study, we investigated the effects of MSHs on DNA damage and DNA methylation of wheat (Triticum aestivum L.) during the seedling growth stage, using the CRED-iPBS (coupled restriction enzyme digestion/inter primer binding site) assay and iPBS analysis to determine DNA methylation status. Exogenous treatment with four MSHs (17-& beta;-estradiol, estrogen, progesterone, and testosterone) was carried out at four different concentrations (0, 0.05, 0.5, and 5 & mu;M). The highest genomic template stability (GTS) value (80%) was observed for 5 & mu;M 17-& beta;-estradiol, 0.5 & mu;M testosterone, and 0.05 & mu;M estrogen, while the lowest value (70.7%) was observed for 5 & mu;M progesterone and 0.5 & mu;M estrogen. The results of the CRED-iPBS analysis conducted on MspI indicate that the 0.05 & mu;M estrogen-treated group had the highest polymorphism value of 40%, while the 5 & mu;M progesterone-treated group had the lowest value of 20%. For HpaII, treatment with 0.5 & mu;M 17-& beta;-estradiol had the highest polymorphism value of 33.3%, while the group treated with 0.05 & mu;M 17-& beta;-estradiol and 0.05 & mu;M progesterone had the lowest value of 19.4%. In conclusion, MSH treatments altered the stability of the genomic template of wheat plants and affected the cytosine methylation status at the seedling growth stage. Upon comprehensive examination of the results, it was seen that the employed methodology successfully detected alterations in cytosine methylation of genomic DNA (gDNA), as well as changes in the pattern of genomic instability.Öğe Modeling Callus Induction and Regeneration in Hypocotyl Explant of Fodder Pea (Pisum sativum var. arvense L.) Using Machine Learning Algorithm Method(Mdpi, 2023) Turkoglu, Aras; Bolouri, Parisa; Haliloglu, Kamil; Eren, Baris; Demirel, Fatih; Isik, Muhammet Islam; Piekutowska, MagdalenaA comprehensive understanding of genetic diversity and the categorization of germplasm is important to effectively identify appropriate parental candidates for the goal of breeding. It is necessary to have a technique of tissue culture that is both effective and reproducible to perform genetic engineering on fodder pea genotypes (Pisum sativum var. arvense L.). In this investigation, the genetic diversity of forty-two fodder pea genotypes was assessed based on their ability of callus induction (CI), the percentage of embryogenic callus by explant number (ECNEP), the percentage of responding embryogenic calluses by explant number (RECNEP), the number of somatic embryogenesis (NSE), the number of responding somatic embryogenesis (RSE), the regeneration efficiency (RE), and the number of regenerated plantlets (NRP). The findings of the ANOVA showed that there were significant differences (p < 0.001) between the genotypes for all in vitro parameters. The method of principal component analysis (PCA) was used to study the correlations that exist between the factors associated with tissue culture. While RE and NRP variables were most strongly associated with Do & gbreve;ruyol, Ova & ccedil;evirme-4, Do & scedil;eli-1, Yolge & ccedil;mez, and Incili-3 genotypes, RECNEP, NSE, RDE, and RECNEP variables were strongly associated with Avc & imath;lar, Ova & ccedil;evirme-3, and Ardahan Merkez-2 genotypes. The in vitro process is a complex multivariate process and more robust analyses are needed for linear and nonlinear parameters. Within the scope of this study, artificial neural network (ANN), random forest (RF), and multivariate adaptive regression spline (MARS) algorithms were used for RE estimation, and these algorithms were also compared. The results that we acquired from our research led us to the conclusion that the employed ANN-multilayer perceptron (ANN-MLP) model (R-2 = 0.941) performs better than the RF model (R-2 = 0.754) and the MARS model (R-2 = 0.214). Despite this, it has been shown that the RF model is capable of accurately predicting RE in the early stages of the in vitro process. The current work is an inquiry regarding the use of RF, MARS, and ANN models in plant tissue culture, and it indicates the possibilities of application in a variety of economically important fodder peas.Öğe Prediction of Grain Yield in Wheat by CHAID and MARS Algorithms Analyses(Mdpi, 2023) Demirel, Fatih; Eren, Baris; Yilmaz, Abdurrahim; Turkoglu, Aras; Haliloglu, Kamil; Niedbala, Gniewko; Bujak, HenrykGenetic 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.Öğe Sodium Azide as a Chemical Mutagen in Wheat (Triticum aestivum L.): Patterns of the Genetic and Epigenetic Effects with iPBS and CRED-iPBS Techniques(Mdpi, 2023) Turkoglu, Aras; Haliloglu, Kamil; Tosun, Metin; Szulc, Piotr; Demirel, Fatih; Eren, Baris; Bujak, HenrykWheat, 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.