المؤلفون: Nacer Bellaloui, Dounya Knizia, Jiazheng Yuan, Qijian Song, Frances Betts, Teresa Register, Earl Williams, Naoufal Lakhssassi, Hamid Mazouz, Henry T. Nguyen, Khalid Meksem, Alemu Mengistu, My Abdelmajid Kassem

المصدر: International Journal of Plant Biology, Vol 15, Iss 2, Pp 452-467 (2024)

مصطلحات موضوعية: soybean, RIL, Forrest, Williams 82, linkage map, micronutrients genes, Science, Biology (General), QH301-705.5

الوصف: Soybean is a major crop in the world and an essential source for minerals, including iron (Fe) and zin (Zn). Deficiency of Fe and Zn in soil and soybean plants result in yield loss and poor seed nutritional qualities. Information on genomic regions and candidate genes controlling seed Fe and Zn accumulation in soybean seed is limited. Therefore, The objective of this research was to identify genetic regions, known as quantitative trait loci (QTL), and candidate genes that control the accumulation of Fe and Zn in soybean mature seeds. A ‘Forrest’ by ‘Williams 82’ (F × W82) recombinant inbred line (RIL) population (n = 306) was used and genotyped using a total of 5405 single nucleotides polymorphism (SNP) markers using Infinium SNP6K BeadChips. A two-year experiment was conducted across two environments: North Carolina in 2018 (NC) and Illinois in 2020 (IL). Only QTL with LOD scores ≥ 2.5, as identified by the composite interval mapping (CIM) method, are reported here. In total, 6 QTL were identified for seed Fe; specifically, 3 QTL (qFe-01-[NC-2018], qFe-02-[NC-2018], and qFe-03-[NC-2018]) were located on chromosomes 1, 2, and 6, respectively, in the NC environment, and 3 QTL (qFe-01-[IL-2020], qFe-02-[IL-2020], and qFe-03-[IL-2020]) were positioned on chromosomes 1, 2, and 12, respectively, in the IL environment. A total of 6 QTL associated with seed Zn were also identified; 4 QTL (qZn-01-[NC-2018]; qZn-02-[NC-2018]; qZn-03-[NC-2018]; and qZn-04-[NC-2018]), respectively on Chr 2, 3, 7, and 19 in NC; and 2 QTL (qZn-01-[IL-2020] and qZn-02-[IL-2020]), respectively, on Chr 5 and 8 in IL. Several functional genes encode Fe- and Zn-proteins, transcription factors, proteins-zinc finger motifs (involved in DNA binding and transcriptional regulation; crosstalk between the regulatory pathways of Zn and Fe transporters) were identified and located within the QTL interval. To our knowledge, and based on the literature available, the QTL identified here on Chr 2 and Chr 6 are novel and were not previously identified. This current research provides a new knowledge of the genetic basis of seed Fe and Zn and the markers associated with QTL. The QTL identified here will contribute to efficient marker assisted selection for higher Fe and Zn content in soybean seeds. The candidate genes and metal-responsive transcription factors may coordinate the expression of both Zn and Fe transporters in response to changes in metal availability, providing new knowledge on minerals uptake and transport mechanisms, allowing for possible genetic engineering application.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2037-0164/15/2/35Test; https://doaj.org/toc/2037-0164Test

الوصول الحر: https://doaj.org/article/50379eeec02341689e513e6834157500Test

المؤلفون: Sandeep Goshika, Khalid Meksem, Khaled R. Ahmed, Naoufal Lakhssassi

المصدر: International Journal of Molecular Sciences, Vol 25, Iss 1, p 106 (2023)

مصطلحات موضوعية: deep neural networks, soybean leaf damage detection, automatic labeling, computer vision, Biology (General), QH301-705.5, Chemistry, QD1-999

الوصف: Soybean (Glycine max (L.) Merr.) is a major source of oil and protein for human food and animal feed; however, soybean crops face diverse factors causing damage, including pathogen infections, environmental shifts, poor fertilization, and incorrect pesticide use, leading to reduced yields. Identifying the level of leaf damage aids yield projections, pesticide, and fertilizer decisions. Deep learning models (DLMs) and neural networks mastering tasks from abundant data have been used for binary healthy/unhealthy leaf classification. However, no DLM predicts and categorizes soybean leaf damage severity (five levels) for tailored pesticide use and yield forecasts. This paper introduces a novel DLM for accurate damage prediction and classification, trained on 2930 near-field soybean leaf images. The model quantifies damage severity, distinguishing healthy/unhealthy leaves and offering a comprehensive solution. Performance metrics include accuracy, precision, recall, and F1-score. This research presents a robust DLM for soybean damage assessment, supporting informed agricultural decisions based on specific damage levels and enhancing crop management and productivity.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/1422-0067/25/1/106Test; https://doaj.org/toc/1661-6596Test; https://doaj.org/toc/1422-0067Test

الوصول الحر: https://doaj.org/article/cbcaeef2210a4aba92ea9ba068289cb7Test

المؤلفون: Nacer Bellaloui, Dounya Knizia, Jiazheng Yuan, Qijian Song, Frances Betts, Teresa Register, Earl Williams, Naoufal Lakhssassi, Hamid Mazouz, Henry T. Nguyen, Khalid Meksem, Alemu Mengistu, My Abdelmajid Kassem

المصدر: Plants, Vol 12, Iss 21, p 3709 (2023)

مصطلحات موضوعية: soybean, RIL, Forrest, Williams 82, linkage map, micronutrients, Botany, QK1-989

الوصف: Understanding the genetic basis of seed Ni and Mo is essential. Since soybean is a major crop in the world and a major source for nutrients, including Ni and Mo, the objective of the current research was to map genetic regions (quantitative trait loci, QTL) linked to Ni and Mo concentrations in soybean seed. A recombinant inbred line (RIL) population was derived from a cross between ‘Forrest’ and ‘Williams 82’ (F × W82). A total of 306 lines was used for genotyping using 5405 single nucleotides polymorphism (SNP) markers using Infinium SNP6K BeadChips. A two-year experiment was conducted and included the parents and the RIL population. One experiment was conducted in 2018 in North Carolina (NC), and the second experiment was conducted in Illinois in 2020 (IL). Logarithm of the odds (LOD) of ≥2.5 was set as a threshold to report identified QTL using the composite interval mapping (CIM) method. A wide range of Ni and Mo concentrations among RILs was observed. A total of four QTL (qNi-01, qNi-02, and qNi-03 on Chr 2, 8, and 9, respectively, in 2018, and qNi-01 on Chr 20 in 2020) was identified for seed Ni. All these QTL were significantly (LOD threshold > 2.5) associated with seed Ni, with LOD scores ranging between 2.71–3.44, and with phenotypic variance ranging from 4.48–6.97%. A total of three QTL for Mo (qMo-01, qMo-02, and qMo-03 on Chr 1, 3, 17, respectively) was identified in 2018, and four QTL (qMo-01, qMo-02, qMo-03, and qMo-04, on Chr 5, 11, 14, and 16, respectively) were identified in 2020. Some of the current QTL had high LOD and significantly contributed to the phenotypic variance for the trait. For example, in 2018, Mo QTL qMo-01 on Chr 1 had LOD of 7.8, explaining a phenotypic variance of 41.17%, and qMo-03 on Chr 17 had LOD of 5.33, with phenotypic variance explained of 41.49%. In addition, one Mo QTL (qMo-03 on Chr 14) had LOD of 9.77, explaining 51.57% of phenotypic variance related to the trait, and another Mo QTL (qMo-04 on Chr 16) had LOD of 7.62 and explained 49.95% of phenotypic variance. None of the QTL identified here were identified twice across locations/years. Based on a search of the available literature and of SoyBase, the four QTL for Ni, identified on Chr 2, 8, 9, and 20, and the five QTL associated with Mo, identified on Chr 1, 17, 11, 14, and 16, are novel and not previously reported. This research contributes new insights into the genetic mapping of Ni and Mo, and provides valuable QTL and molecular markers that can potentially assist in selecting Ni and Mo levels in soybean seeds.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2223-7747/12/21/3709Test; https://doaj.org/toc/2223-7747Test

الوصول الحر: https://doaj.org/article/68ecfb103149484f875841d3c49a3dbbTest

المؤلفون: Naoufal Lakhssassi, Dounya Knizia, Abdelhalim El Baze, Aicha Lakhssassi, Jonas Meksem, Khalid Meksem

المصدر: International Journal of Molecular Sciences, Vol 23, Iss 19, p 11278 (2022)

مصطلحات موضوعية: PLP, THF, SCN resistance, SHMT, soybean, mutational analysis, Biology (General), QH301-705.5, Chemistry, QD1-999

الوصف: The serine hydroxymethyltransferase (SHMT; E.C. 2.1.2.1) is involved in the interconversion of serine/glycine and tetrahydrofolate (THF)/5,10-methylene THF, playing a key role in one-carbon metabolism, the de novo purine pathway, cellular methylation reactions, redox homeostasis maintenance, and methionine and thymidylate synthesis. GmSHMT08 is the soybean gene underlying soybean cyst nematode (SCN) resistance at the Rhg4 locus. GmSHMT08 protein contains four tetrahydrofolate (THF) cofactor binding sites (L129, L135, F284, N374) and six pyridoxal phosphate (PLP) cofactor binding/catalysis sites (Y59, G106, G107, H134, S190A, H218). In the current study, proteomic analysis of a data set of protein complex immunoprecipitated using GmSHMT08 antibodies under SCN infected soybean roots reveals the presence of enriched pathways that mainly use glycine/serine as a substrate (glyoxylate cycle, redox homeostasis, glycolysis, and heme biosynthesis). Root and leaf transcriptomic analysis of differentially expressed genes under SCN infection supported the proteomic data, pointing directly to the involvement of the interconversion reaction carried out by the serine hydroxymethyltransferase enzyme. Direct site mutagenesis revealed that all mutated THF and PLP sites at the GmSHMT08 resulted in increased SCN resistance. We have shown the involvement of PLP sites in SCN resistance. Specially, the effect of the two Y59 and S190 PLP sites was more drastic than the tested THF sites. This unprecedented finding will help us to identify the biological outcomes of THF and PLP residues at the GmSHMT08 and to understand SCN resistance mechanisms.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/1422-0067/23/19/11278Test; https://doaj.org/toc/1661-6596Test; https://doaj.org/toc/1422-0067Test

الوصول الحر: https://doaj.org/article/71c613cdb6b64013b081dc9d71db4ce6Test

المؤلفون: Mariola Usovsky, Naoufal Lakhssassi, Gunvant B. Patil, Tri D. Vuong, Sarbottam Piya, Tarek Hewezi, Robert T. Robbins, Robert M. Stupar, Khalid Meksem, Henry T. Nguyen

المصدر: The Plant Genome, Vol 14, Iss 2, Pp n/a-n/a (2021)

مصطلحات موضوعية: Plant culture, SB1-1110, Genetics, QH426-470

الوصف: Abstract Reniform nematode (RN, Rotylenchulus reniformis Linford & Oliveira) has emerged as one of the most important plant parasitic nematodes of soybean [Glycine max (L.) Merr.]. Planting resistant varieties is the most effective strategy for nematode management. The objective of this study was to identify quantitative trait loci (QTL) for RN resistance in an exotic soybean line, PI 438489B, using two linkage maps constructed from the Universal Soybean Linkage Panel (USLP 1.0) and next‐generation whole‐genome resequencing (WGRS) technology. Two QTL controlling RN resistance were identified—the soybean cyst nematode (SCN, Heterodera glycines) resistance gene GmSNAP18 at the rhg1 locus and its paralog GmSNAP11. Strong association between resistant phenotype and haplotypes of the GmSNAP11 and GmSNAP18 was observed. The results indicated that GmSNAP11 possibly could have epistatic effect on GmSNAP18, or vice versa, with the presence of a significant correlation in RN resistance of rhg1‐a GmSNAP18 vs. rhg1‐b GmSNAP18. Most importantly, our preliminary data suggested that GmSNAP18 and GmSNAP11 proteins physically interact in planta, suggesting that they belong to the same pathway for resistance. Unlike GmSNAP18, no indication of GmSNAP11 copy number variation was found. Moreover, gene‐based single nucleotide polymorphism (SNP) markers were developed for rapid detection of RN or SCN resistance at these loci. Our analysis substantiates synergic interaction between GmSNAP11 and GmSNAP18 genes and confirms their roles in RN as well as SCN resistance. These results could contribute to a better understanding of evolution and subfunctionalization of genes conferring resistance to multiple nematode species and provide a framework for further investigations.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/1940-3372Test

الوصول الحر: https://doaj.org/article/dfa98a1000034b7e8ee0041109d9cfb1Test

المؤلفون: Dounya Knizia, Jiazheng Yuan, Naoufal Lakhssassi, Abdelhalim El Baze, Mallory Cullen, Tri Vuong, Hamid Mazouz, Henry T. Nguyen, My Abdelmajid Kassem, Khalid Meksem

المصدر: Plants, Vol 11, Iss 9, p 1258 (2022)

مصطلحات موضوعية: soybean, RIL, forrest, Williams 82, linkage map, tocopherol, Botany, QK1-989

الوصف: Soybean seeds are rich in secondary metabolites which are beneficial for human health, including tocopherols. Tocopherols play an important role in human and animal nutrition thanks to their antioxidant activity. In this study, the ‘Forrest’ by ‘Williams 82’ (F×W82) recombinant inbred line (RIL) population (n = 306) was used to map quantitative trait loci (QTL) for seed α-tocopherol, β-tocopherol, δ -tocopherol, γ-tocopherol, and total tocopherol contents in Carbondale, IL over two years. Also, the identification of the candidate genes involved in soybean tocopherols biosynthetic pathway was performed. A total of 32 QTL controlling various seed tocopherol contents have been identified and mapped on Chrs. 1, 2, 5, 6, 7, 8, 9, 10, 12, 13, 16, 17, and 20. One major and novel QTL was identified on Chr. 6 with an R2 of 27.8, 9.9, and 6.9 for δ-tocopherol, α-tocopherol, and total tocopherol content, respectively. Reverse BLAST analysis of the genes that were identified in Arabidopsis allowed the identification of 37 genes involved in soybean tocopherol pathway, among which 11 were located close to the identified QTLs. The tocopherol cyclase gene (TC) Glyma.06G084100 is located close to the QTLs controlling δ-tocopherol (R2 = 27.8), α-tocopherol (R2 = 9.96), and total-tocopherol (R2 = 6.95). The geranylgeranyl diphosphate reductase (GGDR) Glyma.05G026200 gene is located close to a QTL controlling total tocopherol content in soybean (R2 = 4.42). The two methylphytylbenzoquinol methyltransferase (MPBQ-MT) candidate genes Glyma.02G002000 and Glyma.02G143700 are located close to a QTL controlling δ-tocopherol content (R2 = 3.57). The two γ-tocopherol methyltransferase (γ-TMT) genes, Glyma.12G014200 and Glyma.12G014300, are located close to QTLs controlling (γ+ß) tocopherol content (R2 = 8.86) and total tocopherol (R2 = 5.94). The identified tocopherol seed QTLs and candidate genes will be beneficial in breeding programs to develop soybean cultivars with high tocopherol contents.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2223-7747/11/9/1258Test; https://doaj.org/toc/2223-7747Test

الوصول الحر: https://doaj.org/article/68e9c6b5d3314915ae80bb079721763aTest

المؤلفون: Dounya Knizia, Jiazheng Yuan, Nacer Bellaloui, Tri Vuong, Mariola Usovsky, Qijian Song, Frances Betts, Teresa Register, Earl Williams, Naoufal Lakhssassi, Hamid Mazouz, Henry T. Nguyen, Khalid Meksem, Alemu Mengistu, My Abdelmajid Kassem

المصدر: Plants, Vol 10, Iss 10, p 2029 (2021)

مصطلحات موضوعية: soybean, RIL, Forrest, Williams 82, linkage map, isoflavone, Botany, QK1-989

الوصف: Isoflavones are secondary metabolites that are abundant in soybean and other legume seeds providing health and nutrition benefits for both humans and animals. The objectives of this study were to construct a single nucleotide polymorphism (SNP)-based genetic linkage map using the ‘Forrest’ by ‘Williams 82’ (F×W82) recombinant inbred line (RIL) population (n = 306); map quantitative trait loci (QTL) for seed daidzein, genistein, glycitein, and total isoflavone contents in two environments over two years (NC-2018 and IL-2020); identify candidate genes for seed isoflavone. The FXW82 SNP-based map was composed of 2075 SNPs and covered 4029.9 cM. A total of 27 QTL that control various seed isoflavone traits have been identified and mapped on chromosomes (Chrs.) 2, 4, 5, 6, 10, 12, 15, 19, and 20 in both NC-2018 (13 QTL) and IL-2020 (14 QTL). The six QTL regions on Chrs. 2, 4, 5, 12, 15, and 19 are novel regions while the other 21 QTL have been identified by other studies using different biparental mapping populations or genome-wide association studies (GWAS). A total of 130 candidate genes involved in isoflavone biosynthetic pathways have been identified on all 20 Chrs. And among them 16 have been identified and located within or close to the QTL identified in this study. Moreover, transcripts from four genes (Glyma.10G058200, Glyma.06G143000, Glyma.06G137100, and Glyma.06G137300) were highly abundant in Forrest and Williams 82 seeds. The identified QTL and four candidate genes will be useful in breeding programs to develop soybean cultivars with high beneficial isoflavone contents.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2223-7747/10/10/2029Test; https://doaj.org/toc/2223-7747Test

الوصول الحر: https://doaj.org/article/907af698cf6b4b9499d773bc58690e3aTest

المؤلفون: Oussama Badad, Naoufal Lakhssassi, Nabil Zaid, Abdelhalim El Baze, Younes Zaid, Jonas Meksem, David A Lightfoot, Huseyin Tombuloglu, El Houcine Zaid, Turgay Unver, Khalid Meksem

المصدر: Plants, Vol 10, Iss 7, p 1405 (2021)

مصطلحات موضوعية: olive epigenomic, differential methylation, olive genetic diversity, Olea europaea ssp. europaea var. sylvestris, aromatic volatile compounds, Botany, QK1-989

الوصف: Secondary metabolites are particularly important to humans due to their pharmaceutical properties. Moreover, secondary metabolites are key compounds in climate change adaptation in long-living trees. Recently, it has been described that the domestication of Olea subspecies had no major selection signature on coding variants and was mainly related to changes in gene expression. In addition, the phenotypic plasticity in Olea subspecies was linked to the activation of transposable elements in the genes neighboring. Here, we investigated the imprint of DNA methylation in the unassigned fraction of the phenotypic plasticity of the Olea subspecies, using methylated DNA immuno-precipitation sequencing (MeDIP-seq) for a high-resolution genome-wide DNA methylation profiling of leaves and fruits during fruit development in wild and cultivated olives from Turkey. Notably, the methylation profiling showed a differential DNA methylation in secondary metabolism responsible for the sensory quality of olive oil. Here, we highlight for the first time the imprint of DNA methylation in modulating the activity of the Linoleate 9S lipoxygenase in the biosynthesis of volatile aromatic compounds. Unprecedently, the current study reveals the methylation status of the olive genome during fruit ripening.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2223-7747/10/7/1405Test; https://doaj.org/toc/2223-7747Test

الوصول الحر: https://doaj.org/article/f45d133770014cf1b8b07e71d831138aTest

المؤلفون: Naoufal Lakhssassi, Valéria Stefania Lopes-Caitar, Dounya Knizia, Mallory A. Cullen, Oussama Badad, Abdelhalim El Baze, Zhou Zhou, Mohamed G. Embaby, Jonas Meksem, Aicha Lakhssassi, Pengyin Chen, Amer AbuGhazaleh, Tri D. Vuong, Henry T. Nguyen, Tarek Hewezi, Khalid Meksem

المصدر: Cells, Vol 10, Iss 5, p 1245 (2021)

مصطلحات موضوعية: high oleic acid, alternative fatty acid pathway, GmFAD2-2 subfamily, TILLING-by-Sequencing+, subfunctionalization, neofunctionalization, Cytology, QH573-671

الوصف: Soybean is the second largest source of oil worldwide. Developing soybean varieties with high levels of oleic acid is a primary goal of the soybean breeders and industry. Edible oils containing high level of oleic acid and low level of linoleic acid are considered with higher oxidative stability and can be used as a natural antioxidant in food stability. All developed high oleic acid soybeans carry two alleles; GmFAD2-1A and GmFAD2-1B. However, when planted in cold soil, a possible reduction in seed germination was reported when high seed oleic acid derived from GmFAD2-1 alleles were used. Besides the soybean fatty acid desaturase (GmFAD2-1) subfamily, the GmFAD2-2 subfamily is composed of five members, including GmFAD2-2A, GmFAD2-2B, GmFAD2-2C, GmFAD2-2D, and GmFAD2-2E. Segmental duplication of GmFAD2-1A/GmFAD2-1B, GmFAD2-2A/GmFAD2-2C, GmFAD2-2A/GmFAD2-2D, and GmFAD2-2D/GmFAD2-2C have occurred about 10.65, 27.04, 100.81, and 106.55 Mya, respectively. Using TILLING-by-Sequencing+ technology, we successfully identified 12, 8, 10, 9, and 19 EMS mutants at the GmFAD2-2A, GmFAD2-2B, GmFAD2-2C, GmFAD2-2D, and GmFAD2-2E genes, respectively. Functional analyses of newly identified mutants revealed unprecedented role of the five GmFAD2-2A, GmFAD2-2B, GmFAD2-2C, GmFAD2-2D, and GmFAD2-2E members in controlling the seed oleic acid content. Most importantly, unlike GmFAD2-1 members, subcellular localization revealed that members of the GmFAD2-2 subfamily showed a cytoplasmic localization, which may suggest the presence of an alternative fatty acid desaturase pathway in soybean for converting oleic acid content without substantially altering the traditional plastidial/ER fatty acid production.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2073-4409/10/5/1245Test; https://doaj.org/toc/2073-4409Test

الوصول الحر: https://doaj.org/article/a9a63ab689b34d63893a739b100dbbf4Test

المؤلفون: Naoufal Lakhssassi, Zhou Zhou, Mallory A. Cullen, Oussama Badad, Abdelhalim El Baze, Oumaima Chetto, Mohamed G. Embaby, Dounya Knizia, Shiming Liu, Leandro G. Neves, Khalid Meksem

المصدر: International Journal of Molecular Sciences, Vol 22, Iss 8, p 4219 (2021)

مصطلحات موضوعية: TILLING by target capture sequencing, TILLING-by-sequencing+, oil biosynthesis pathway, fatty acid desaturases, GmSACPD-C, GmFAD2-1A/1B, Biology (General), QH301-705.5, Chemistry, QD1-999

الوصف: Reverse genetic approaches have been widely applied to study gene function in crop species; however, these techniques, including gel-based TILLING, present low efficiency to characterize genes in soybeans due to genome complexity, gene duplication, and the presence of multiple gene family members that share high homology in their DNA sequence. Chemical mutagenesis emerges as a genetically modified-free strategy to produce large-scale soybean mutants for economically important traits improvement. The current study uses an optimized high-throughput TILLING by target capture sequencing technology, or TILLING-by-Sequencing+ (TbyS+), coupled with universal bioinformatic tools to identify population-wide mutations in soybeans. Four ethyl methanesulfonate mutagenized populations (4032 mutant families) have been screened for the presence of induced mutations in targeted genes. The mutation types and effects have been characterized for a total of 138 soybean genes involved in soybean seed composition, disease resistance, and many other quality traits. To test the efficiency of TbyS+ in complex genomes, we used soybeans as a model with a focus on three desaturase gene families, GmSACPD, GmFAD2, and GmFAD3, that are involved in the soybean fatty acid biosynthesis pathway. We successfully isolated mutants from all the six gene family members. Unsurprisingly, most of the characterized mutants showed significant changes either in their stearic, oleic, or linolenic acids. By using TbyS+, we discovered novel sources of soybean oil traits, including high saturated and monosaturated fatty acids in addition to low polyunsaturated fatty acid contents. This technology provides an unprecedented platform for highly effective screening of polyploid mutant populations and functional gene analysis. The obtained soybean mutants from this study can be used in subsequent soybean breeding programs for improved oil composition traits.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/1422-0067/22/8/4219Test; https://doaj.org/toc/1661-6596Test; https://doaj.org/toc/1422-0067Test

الوصول الحر: https://doaj.org/article/52b916f9845145ccb68e83d747a56551Test