Browsing by Author "Kasapoglu, Ayse Gul"

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Characterization of Two-Component System Gene (TCS) in Melatonin-Treated Common Bean Under Salt and Drought Stress
(Springer, 2023) Kasapoglu, Ayse Gul; Ilhan, Emre; Aydin, Murat; Yigider, Esma; Inal, Behcet; Buyuk, Ilker; Agar, Guleray
The two-component system (TCS) generally consists of three elements, namely the histidine kinase (HK), response regulator (RR), and histidine phosphotransfer (HP) gene families. This study aimed to assess the expression of TCS genes in P. vulgaris leaf tissue under salt and drought stress and perform a genome-wide analysis of TCS gene family members using bioinformatics methods. This study identified 67 PvTCS genes, including 10 PvHP, 38 PvRR, and 19 PvHK, in the bean genome. PvHK2 had the maximum number of amino acids with 1261, whilst PvHP8 had the lowest number with 87. In addition, their theoretical isoelectric points were between 4.56 (PvHP8) and 9.15 (PvPRR10). The majority of PvTCS genes are unstable. Phylogenetic analysis of TCS genes in A. thaliana, G. max, and bean found that PvTCS genes had close phylogenetic relationships with the genes of other plants. Segmental and tandem duplicate gene pairs were detected among the TCS genes and TCS genes have been subjected to purifying selection pressure in the evolutionary process. Furthermore, the TCS gene family, which has an important role in abiotic stress and hormonal responses in plants, was characterized for the first time in beans, and its expression of TCS genes in bean leaves under salt and drought stress was established using RNAseq and qRT-PCR analyses. The findings of this study will aid future functional and genomic studies by providing essential information about the members of the TCS gene family in beans.
Evaluation of Genetic Diversity Using IPBS-Scot Marker Methods in Native Hawthorn Genetic Resources and Species Identification by Using DNA Barcoding Method
(C M B Assoc, 2023) Sagbas, Halil Ibrahim; Ercisli, Sezai; Aydin, Murat; Ilhan, Emre; Aydinyurt, Recep; Kasapoglu, Ayse Gul; Polat, Yakup
Hawthorn is an important medicinal plant that spreads around the world and is used in traditional Chinese medicine. Its flowers and leaves contain flavonoids, vitamins, organic acids and essential oils. Its fruit is consumed as fresh and dried and is an important plant for human health. In this study, iPBS (Inter Primer Binding Site) and SCoT (Start Codon Target Polymorphism) markers were used to analyze genetic variation among 101 hawthorn genotypes collected from coruh Valley, Turkiye and ITS markers were used for DNA barcoding. Ten iPBS primers were used and a total of 400 alleles were identified from ten iPBS primers with an average of 40 alleles. PIC values ranged from 0.239 (iPBS 2387) to 0.272 (iPBS 2244). Twenty SCoT pri-mers were used and have an average of 50.05 alleles. The PIC values of the primers ranged from 0.251 (SCoT 2) to 0.297 (SCoT 34). For the DNA barcoding study, it was confirmed that the correct region was amplified and sequenced. The genotypes we used in the study matched 14 different accession numbers by searching a BLASTN in the NCBI. NCBI similarity rates of hawthorn genotypes are between 90.83% and 100%. The study emphasizes the genetic diversity of hawthorn grown from seed and the importance of preserving plant genetic resources.
Evaluation of Sodium Azide as a Chemical Mutagen in Developing Cold-Tolerant Quinoa (Chenopodium Quinoa Willd.) Lines
(Springer Int Publ Ag, 2023) Ilhan, Emre; Kasapoglu, Ayse Gul; Turkoglu, Aras; Aygoren, Ahmed Sidar; Muslu, Selman; Aydin, Murat; Haliloglu, Kamil
Quinoa (Chenopodium quinoa Willd.) is an allotetraploid grain crop belonging to the Amaranthaceae family. With a nutritionally well-balanced nature and rich oil, protein, vitamin, carbohydrate, and amino acid content, it has various health benefits. Hybridization programs to induce genetic variation are difficult processes in quinoa plants; however, the introduction of variations through mutation to obtain promising genotypes is much easier. In this research, quinoa cultivar Atlas seeds were exposed to different doses and application durations of sodium azide (NaN3) were cultivated in pots. The present findings revealed distinctive differences in all studied traits between native and mutant plants. NaN3 mutagenesis caused genetic variations that were categorized based on studied traits and RAPD data, which led to two distinct groups during M-2. It was noted that 2 mM + 3 h NaN3 treatments had the greatest effects on genetic variations and germination parameters. Present findings were also confirmed by molecular analysis. This research will provide new insights into molecular breeding programs to be conducted to improve present quinoa genotypes and produce better genotypes.
Expression Analysis and Characterization of the CPP Gene Family of Melatonin-Treated Common Bean Cultivars Under Different Abiotic Stresses
(Elsevier, 2023) Rakhimzhanova, Akgul; Kasapoglu, Ayse Gul; Sapakova, Aigul; Ilhan, Emre; Zharmukhametova, Raushan; Turan, Murat; Ciltas, Abdulkadir
Cysteine-rich polycomb-like protein (CPP) transcription factors, which are prevalent in plants and animals but absent in prokaryotes and fungi, play a significant role in response to various abiotic stresses and the reg-ulation of cell division and development of reproductive tissues in plants. Plants can not avoid environmental challenges because they are soil-bound organisms. Therefore, they activate physiological, pharmacological, and molecular stress defence mechanisms. Among the molecular defence systems is the repression or stimu-lation of the expression of specific genes or gene families by particular transcription factors. Identification of these gene families is of great importance. The CPP gene family has been identified in some plants but not in beans. This study aimed to identify and characterize the CPP genes in the P. vulgaris genome and investigate them using various bioinformatics tools on a genome-wide scale. Expression levels of melatonin and stress factors PvCPP genes were determined by qRT-PCR analysis. Seven CPP genes have been identified in the Pha-seolus vulgaris genome. Except for the PvCPP7 gene, the other PvCPP genes contain two CXC domains. The CPP genes of Phaseolus vulgaris, Arabidopsis thaliana, and Glycine max were analyzed for their phylogenetic rela-tionships and were separated into three distinct groups. In qRT-PCR analysis, 5 CPP genes were studied and their expression levels in stress and melatonin applications were determined and differences in expression levels of PvCPP genes were observed in all treatments. The information generated from this study will serve as a foundation for future functional genomics research by understanding the structure and function of CPP genes in bean genotypes.& COPY; 2023 SAAB. Published by Elsevier B.V. All rights reserved.
Genetic Diversity Analysis of Lesser Burnet (Sanguisorba Minor Scop.) Genotypes Collected from Erzurum Province, Türkiye Using Inter-Simple Sequence Repeat (ISSR) Markers
(Springer, 2025) Karatas, Rufayi; Kasapoglu, Ayse Gul; Gullap, Mehmet Kerim; Ilhan, Emre; Aydin, Murat
This research was conducted over a three-year period from 2021 to 2023, utilizing the ISSR marker system to determine the kinship relationships among populations obtained from seeds of lesser burnet (Sanguisorba minor Scop.) collected from the natural pasture areas of the Erzurum region. The study revealed genetic diversity among 42 burnet genotypes, including 40 genotypes and 2 control cultivars (B & uuml;nyan 80 and Alt & imath;nova). The work with ISSR primers yielded PIC values ranging from 0.24 to 0.34, with an average of 0.30. The highest PIC value (0.34) was obtained from the UBC811 primer, while the lowest value (0.24) was from the UBC812 primer. Among the ISSR primers, UBC811 showed the highest Na, Ne, I, and h values with 2, 1.63, 0.54, and 0.37, respectively, whereas the UBC812 primer displayed the lowest Na, Ne, I, and h values with 2, 1.40, 0.42, and 0.26, respectively. When the results were evaluated as a whole, it was determined that there was no relationship among the collected lesser burnet (S. minor Scop.) populations solely in terms of geographical isolation. Additionally, we recorded that almost all of the 40 collected genotypes significantly differed from the commercial cultivars used as controls, indicating potential for the development of new cultivars.
Genetic Diversity and Population Structure of Barley Cultivars Released in Turkey and Bulgaria Using IPBS-Retrotransposon and SCOT Markers
(Ankara Univ, Fac Agriculture, 2022) Gungor, Huseyin; Ilhan, Emre; Kasapoglu, Ayse Gul; Filiz, Ertugrul; Hossein-Pour, Arash; Valchev, Dragomir; Dumlupinar, Ziya
To improve quantitative traits, it is essential to acknowledge genetic structure and diversity of the crop plants. In this study, 54 barley cultivars released from 1963 to date by different institutes in both Turkey and Bulgaria were screened with 18 iPBS and four SCoT markers to evaluate population structure and genetic diversity. According to the results, while total polymorphic band numbers was identified as 560, the polymorphic ones were found as 530 (438 and 92 amplified bands for iPBS and SCoT markers, respectively). In addition, the average polymorphic band number was found as 24.09. While the average polymorphism information content (PIC) value was 0.48, the average PIC value was 0.48 for iPBS and 0.48 for SCoT markers. The highest PIC value was determined as 0.50. The highest effective number of alleles, Shannon's information index, and Nei's genetic diversity were detected from the iPBS2271 marker at 1.61, 0.52 and 0.35, respectively among the iPBS markers while the highest values were obtained from SCoT-71 marker as 1.55, 0.32 and 0.48, respectively. As a result of a distribution of the 530 amplified bands in 54 barley cultivars, structure analysis showed that the subpopulations in the barley cultivars as a value of k=5. The average expected heterozygosity and fixation indices were identified as 0.234 and 0.322, respectively. Based on DICE similarity index, Marti and Zahir cultivars were found the most similar barley cultivars with 75% genetic similarity, whereas Ozdemir and Karatay 94 and Tosunpasa and Konevi cultivars were found 73% similar. On the other hand, Bayrak and Avci-2002 were found the most diverse cultivars with 19.9% genetic similarity. As a result, the barley cultivars released in Turkey and Bulgaria were found varying and, the genetic diversity and statistics index analysis indicated that iPBS and SCoT markers are powerful markers to perform genetic diversity analysis.
Genome-Wide Analysis and Characterization of Eucalyptus Grandis TCP Transcription Factors
(Ankara University, Faculty of Agriculture, 2023) Ilhan, Emre; Kasapoglu, Ayse Gul; Muslu, Selman; Aygoren, Ahmed Sidar; Aydin, Murat
Teosinte branched/Cycloidea/Proliferating cell factors (TCPs), a small transcription gene family, serve in developmental processes such as branching, flowering, and growth of plants. In this study, the TCP transcription gene family of eucalyptus, which is considered important for its medicinal and industrial uses, was bioinformatically investigated. A total of 16 Eucalyptus grandis TCP (Egra-TCP) genes were found to be distributed on chromosomes 1, 2, 4, 6, 7, 9, 10 and 11. Several segmentally-duplicated gene couples including Egra-TCP-7/ Egra-TCP-11,-13 and-16, Egra-TCP-6/Egra-TCP-12 and-15, Egra-TCP-12/ Egra-TCP-15 and Egra-TCP-11/Egra-TCP-13 were discovered. Egra-TCPs were divided into three main clades based on phylogenetic analysis, motif, and gene structure. While Egra-TCP-10 has the highest molecular weight with 47.19 kDa, the lowest was Egra-TCP-1 with 21.68 kDa. Twelve Egra-TCP genes were found to have no introns, while the Egra-TCP-7,-15, and-16 genes had a single intron. The orthologous relationships among E. grandis/Arabidopsis thaliana and E. grandis/Vitis vinifera were identified through a synteny analysis. Digital gene expression profiles of Egra-TCP genes in tissues such as xylem, phloem, shoot tips, young and mature leaf revealed a high expression pattern. The findings of this study contributes to existing knowledge in the biotechnology field by providing contributing to our understanding of the molecular basis of the TCP gene family in the eucalyptus plant.
Genome-Wide Analysis and Characterization of Sabath Gene Family in Phaseolus Vulgaris Genotypes Subject to Melatonin Under Drought and Salinity Stresses
(Springer, 2023) Aygoren, Ahmed Sidar; Gunes, Ebru; Muslu, Selman; Kasapoglu, Ayse Gul; Yigider, Esma; Aydin, Murat; Ilhan, Emre
The SABATH gene family is a plant-specific class of small molecule methyltransferases. The first goal of this study was to conduct a genome-wide analysis and in silico characterization of SABATH gene family members in Phaseolus vulgaris. The second goal was to see how the members of the SABATH gene family react to salt and drought stress, as well as their expression patterns when melatonin is added to the mix. Within the scope of the study, 18 Pvul-SABATH proteins were discovered in the Phaseolus vulgaris genome using various in silico methods. These proteins ranged in size from 30.18 to 42.73 kDa and were made up of 268 to 387 amino acids. The Pvul-SABATH proteins had isoelectric points ranging from 4.95 (Pvul-SABATH-8) to 6.08 (Pvul-SABATH-2 and -3). The Pvul-SABATH genes were found to have at least three but no more than four exons. According to the results of the phylogenetic analysis, Pvul-SABATH proteins were found to be clustered in three major groups with Arabidopsis thaliana, Glycine max species, and various methyltransferase members. There was segmental duplication between the Pvul-SABATH-1 and Pvul-SABATH-5 genes, as well as the Pvul-SABATH-10 and Pvul-SABATH-14 genes. By comparing the expression level of Pvul-SABATH genes, their different expression levels depending on the bean cultivars were discovered. Moreover, these findings suggested that Pvul-SABATH genes may play a role in the growth and development process of the plants in response to various biotic and abiotic stresses. The findings of this study, which is the first to look at the SABATH gene family in Phaseolus vulgaris, are thought to be beneficial in plant biotechnology and molecular biology.
Genome-Wide Analysis of Glutathione S-Transferase Gene Family in P. Vulgaris Under Drought and Salinity Stress
(Springer, 2024) Muslu, Selman; Kasapoglu, Ayse Gul; Gunes, Ebru; Aygoren, Ahmed Sidar; Yigider, Esma; Ilhan, Emre; Aydin, Murat
The enzymes known as glutathione S-transferases (GST), which are present in many evolved organisms, are essential for the defense against reactive oxygen species. GSTs have a role in the development of defenses against biotic and abiotic challenges, especially defending plants from various stresses such as drought, salinity, and heavy metal. This study uses a genome-wide investigation of the GST gene family in Phaseolus vulgaris to pinpoint several distinctive traits. 55 Pv-GST proteins have been identified in P. vulgaris. The molecular weights of these proteins range from 15.02 kDa to 47.99 kDa; the range of amino acid numbers 132 to 420 and the range of theoretical isoelectric points 5.03 to 9.61 were identified. Pv-GST genes are estimated to have at least 2 and a maximum of 10 exons, with an average of 4 exons. Phylogenetic analysis was performed with GST proteins from Arabidopsis thaliana, Glycine max, and Phaseolus vulgaris species, and subfamilies of these GSTs were identified. Using RNAseq data, the expression profiles of Pv-GST genes in leaf tissue of common bean during drought and salinity stress were identified. Using the obtained sequence data, primers for qRT-PCR were designed. Changes in the expression profiles of GST genes caused by salt and drought stress and melatonin treatments in two different common bean cultivars were determined by qRT-PCR experiments. Under drought and salt stress, the expression levels of GSTs decreased, while melatonin treatment with few exceptions increased these expression levels. As a result of this study, it was observed that Pv-GST genes may play a role in the growth and development of bean and may be involved in the response to abiotic stresses. Moreover, the results of this study will provide a basis for functional gene research and the expression profiles and qRT-PCR results under different stress conditions in common bean will provide a valuable contribution to the understanding of the functions of the GST gene family.
Genome-Wide Analysis of Mir172-Mediated Response to Heavy Metal Stress in Chickpea (Cicer Arietinum L.): Physiological, Biochemical, and Molecular Insights
(BMC, 2024) Ucar, Sumeyra; Yaprak, Esra; Yigider, Esma; Kasapoglu, Ayse Gul; Oner, Burak Muhammed; Ilhan, Emre; Aydin, Murat
BackgroundChickpea (Cicer arietinum L.), a critical diploid legume in the Fabaceae family, is a rich source of protein, vitamins, and minerals. However, heavy metal toxicity severely affects its growth, yield, and quality. MicroRNAs (miRNAs) play a crucial role in regulating plant responses to both abiotic and biotic stress, including heavy metal exposure, by suppressing the expression of target genes. Plants respond to heavy metal stress through miRNA-mediated regulatory mechanisms at multiple physiological, biochemical, and molecular levels. Although the Fabaceae family is well represented in miRNA studies, chickpeas have been notably underrepresented. This study aimed to investigate the effects of heavy metal-induced stress, particularly from 100 mu M concentrations of cadmium (Cd), chromium (Cr), nickel (Ni), lead (Pb), and 30 mu M arsenic (As), on two chickpea varieties: ILC 482 (sensitive) and Azkan (tolerant). The assessment focused on physiological, biochemical, and molecular parameters. Furthermore, a systematic characterization of the miR172 gene family in the chickpea genome was conducted to better understand the plant's molecular response to heavy metal stress.ResultsVariance analysis indicated significant effects of genotype (G), treatment (T), and genotype-by-treatment (GxT) interactions on plant growth, physiological, and biochemical parameters. Heavy metal stress negatively impacted plant growth in chickpea genotypes ILC 482 and Azkan. A reduction in chlorophyll content and relative leaf water content was observed, along with increased cell membrane damage. In ILC 482, the highest hydrogen peroxide (H2O2) levels in shoot tissue were recorded under As, Cd, and Ni treatments, while in Azkan, peak levels were observed with Pb treatment. Malondialdehyde (MDA) levels in root tissue were highest in ILC 482 under Cd and Ni exposure and in Azkan under As, Cr, and Cd treatments. Antioxidant enzyme activities, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX), were significantly elevated under heavy metal stress in both genotypes. Gene expression analysis revealed upregulation of essential antioxidant enzyme genes, such as SOD, CAT, and APX, with APX showing notable increases in both shoot and root tissues compared to the control. Additionally, seven miR172 genes (miR172a, miR172b, miR172c, miR172d, miR172e, miR172f, and miR172g) were identified in the chickpea genome, distributed across five chromosomes. All genes exhibited conserved hairpin structures essential for miRNA functionality. Phylogenetic analysis grouped these miR172 genes into three clades, suggesting strong evolutionary conservation with other plant species. The expression analysis of miR172 and its target genes under heavy metal stress showed varied expression patterns, indicating their role in enhancing heavy metal tolerance in chickpea.ConclusionsHeavy metal stress significantly impaired plant growth and physiological and biochemical parameters in chickpea genotypes, except for cell membrane damage. The findings underscore the critical role of miR172 and its target genes in modulating chickpea's response to heavy metal stress. These insights provide a foundational understanding for developing stress-tolerant chickpea varieties through miRNA-based genetic engineering approaches.
Genome-Wide Characterization of the Gpat Gene Family in Bean (Phaseolus Vulgaris L.) and Expression Analysis Under Abiotic Stress and Melatonin
(Springer, 2024) Kasapoglu, Ayse Gul; Muslu, Selman; Aygoren, Ahmed Sidar; Oner, Burak Muhammed; Gunes, Ebru; Ilhan, Emre; Aydin, Murat
GPAT (sn-Glycerol-3-phosphate-1-O-acyltransferase) is a key enzyme in the biosynthesis of membrane lipids. Two kinds of GPATs are involved in the production of glycerolipids. While sn-1-GPATs are responsible for creating lysophosphatidic acid, sn-2-GPATs generate monoacylglycerol. GPAT genes of beans (Phaseolus vulgaris) have never been investigated before. This study aimed to assess the expression profiles of GPAT genes in common bean leaves under drought and salt stress and apply bioinformatics tools to the genome-wide analysis of GPAT gene family members. In the P. vulgaris genome, PvGPAT genes with molecular weights ranging from 50.2 to 60.4 kDa, amino acid counts ranging from 376 to 539, theoretical isoelectric points ranging from 8.41 to 9.46, and instability indices ranging from 31.84 to 51.38 were found. Based on the gene structure analysis, the expected number of exons and introns was between 2 and 12. The GPAT proteins from Arabidopsis thaliana, Glycine max, Vitis vinifera, Cicer arietinum, Lens culinaris, and P. vulgaris were analyzed phylogenetically. GPAT proteins of G. max and A. thaliana are evolutionarily related to those of beans. The PvGPAT4/PvGPAT10 and PvGPAT7/PvGPAT12 gene pairs were detected to be segmentally duplicated. In this work, qRT-PCR was performed on six PvGPAT genes, and the effects of salt, drought, and melatonin treatments on bean GPAT genes were studied. The role of melatonin in the response to salt and drought stress was investigated at the gene expression level. The findings of the present study will shed light on GPAT gene family members in beans and provide insight for future functional research.
Identification and Characterization of Common Bean Ofp Gene Family Under Environmental Stresses
(Springer, 2025) Ucar, Sumeyra; Muslu, Selman; Gunes, Ebru; Kasapoglu, Ayse Gul; Aygoren, Ahmed Sidar; Yaprak, Esra; Yildirim, Ertan
The OVATE FAMILY PROTEIN (OFP) is a remarkable class of plant-specific transcription factors essential for plant growth and development. Common bean (Phaseolus vulgaris L.), the most commonly cultivated legume, was the focus of this study to evaluate the expression of OVATE genes in response to salt and drought stress, along with a genome-wide analysis of the OVATE gene family using bioinformatics tools. The study also involves comprehensive bioinformatics analyses to identify the 22 members of the Pvul-OFP gene family's conserved domains, chromosomal locations, evolutionary relationships, genetic structures, and expression patterns. Additionally, the expression levels of OFP genes in plants treated with melatonin, which has an antioxidant effect against salt and drought stress, were examined using qRT-PCR. Phylogenetic analysis revealed that Pvul-OFP genes are similar to OFP gene families found in Arabidopsis thaliana and Glycine max. Moreover, segmental duplication was detected between Pvul-OFP gene pairs. In addition, while the Pvul-OFP-2 gene had the highest expression level under salt stress, the highest expression level was determined in the Pvul-OFP-22 gene under drought stress. Moreover, it was determined that the expression levels of Pvul-OFP-7 and Pvul-OFP-20 genes did not change in both stress groups compared to the control group. The qRT-PCR results showed significant changes in the expression levels of OFP genes across different varieties and in shoot tissue. This comprehensive analysis will aid future studies associated with the functional validation of OFP genes and contribute to developing cultivars resistant to stress conditions.
Investigation of Biochemical and Molecular Changes in Wheat Genotypes Under Alkaline Salt Stress
(Springer Heidelberg, 2024) Kizilkaya, Damla; Ilhan, Emre; Aydin, Murat; Kasapoglu, Ayse Gul; Yigider, Esma; Turkoglu, Aras; Haliloglu, Kamil
Wheat, a glycophyte plant that is the main staple food for the majority of the world's population, is considerably sensitive to salinity and alkalinity stress. The goal of this study was to investigate antioxidant enzyme activity and elemental analyses to determine the response of three wheat genotypes to alkaline stress during the early development period as well as to examine DNA damage caused by alkaline stress using the inter simple sequence repeats (ISSR) molecular markers. The results demonstrated that NaHCO3 treatments affected the amount and ratio of Na and K in all genotypes' roots and leaves. Furthermore, NaHCO3 treatment had a significant impact on H2O2 contents and malondialdehyde (MDA) levels of genotypes which exhibited varying abilities to reduce reactive oxygen species-induced damage and stress severity affected enzymatic antioxidant systems (SOD, CAT, and POD activities). The ISSR results revealed that genomic template stability rates decreased in response to alkaline stress. Overall, the data revealed significant genotype by alkaline salt exposure interaction, indicating that genetic response to alkaline salt stress may be different with respect to cultivars. In our study, cv. & Ccedil;etinel 2000 had more enzymatic and non-enzymatic activity compared to cv. Aytin-98 and Tir genotypes under concentrations NaHCO3 concentration, while latter two genotypes varied responses under the different concentrations. Our research also showed that genetic variability does exist in wheat for alkaline salt tolerant/resistant genes, further research using transcriptomic techniques is required to establish the gene expression profiles of wheat genotypes under such stress conditions to assess the genetic information about the related genes.
Micropropagation and Genetic Fidelity of Rheum Ribes L.: A Threatened and Endemic Medicinal Herb from the Turkish Eastern Anatolia Region
(Springer, 2025) Aldaif, Muhammed; Ucar, Sumeyra; Yigider, Esma; Aydin, Murat; Kasapoglu, Ayse Gul; Ilhan, Emre
Rheum ribes L., an endemic medicinal herb of Eastern Anatolia, faces serious conservation concerns due to habitat degradation, seed dormancy, and overharvesting. To overcome these challenges, a robust and reproducible in vitro regeneration system was developed using embryo axis, hypocotyl, and cotyledon explants via direct and indirect organogenesis as well as somatic embryogenesis. The highest germination rate (88.8%) and shortest germination time were obtained using 1/2 MS medium supplemented with 1.75 mg/L GA(3) and 1.0 mg/L IBA in mature embryos. The micropropagation experiments were conducted in two stages. In the first stage, embryo axis, cotyledon, and hypocotyl explants were cultured using 36 different methods that included various culture conditions and types of plant growth regulators at differing concentrations. In the second stage, calli from these explants were cultured in the most effective medium (method 30) for callus production, and the explants were subsequently cultured in eight different regeneration media based on MS medium supplemented with varying concentrations of plant growth regulators. Callus induction reached 100% under dark conditions in MS medium containing 0.5 mg/L BAP, 0.5 mg/L KIN, and 1.0 mg/L 2,4-D. Among 36 regeneration protocols, method 13 (3.0 mg/L BAP, 0.2 mg/L NAA, 1.0 mg/L GA(3)) yielded the highest plant regeneration (1.15 plantlets/explant) in embryo axis and hypocotyl explants. In the second stage, the highest indirect somatic embryogenesis (ISER) rate (55.6%) was observed in cotyledon-derived callus (C-callus). In contrast, the highest indirect organogenesis (IOR) (28.1%) was found in embryo axis-derived callus (E-callus). The maximum number of plantlets per explant (PPEN: 1.08) was achieved in MS medium containing 2.0 mg/L BAP, 2.0 mg/L KIN, and 1.0 mg/L GA(3). Genetic fidelity of 15 regeneration systems was assessed using 20 highly polymorphic SCoT primers. Similarity coefficients between regenerated plants and the mother plant ranged from 0.904 to 0.991, with the highest genetic stability observed in plantlets derived via direct somatic embryogenesis. This optimized regeneration protocol is a powerful tool for the large-scale clonal propagation and ex-situ conservation of R. ribes, enabling both genetic preservation and sustainable biotechnological utilization of this pharmacologically valuable species.