SCoT Primers Research Articles

Effect of meta-Topolin on morphological, physiochemical, and molecular dynamics during in vitro regeneration of Salix tetrasperma Roxb.

An efficient in vitro propagation protocol has been established for a valuable medicinal plant, Salix tetrasperma using mature nodal explants. The investigation aimed to observe the influence of various combinations and concentrations of cytokinins (mT, BA, and Kn) and auxins (NAA, IAA, and IBA) on regeneration potential using the Murashige and Skoog (MS) medium. Among individual cytokinin treatments, 5.0 µM mT resulted highest response of 92% with maximum shoot number (11.6 ± 0.08) per explant and shoot length (4.5 ± 0.22 cm) after 12 weeks of culture. However, synergistic treatment of mT (5.0 µM) and NAA (0.5 µM) further improved proliferation with (21.3 ± 0.40) shoots per explant and (6.9 ± 0.13 cm) shoot length in 96% cultures after 12 weeks of incubation. Rooting from in vitro raised microshoots was achieved on ½ MS medium supplemented with various concentrations of low-dose auxins. The highest number of roots (10.4 ± 0.20) per shoot with mean root length (5.7 ± 0.11 cm) with maximum rooting frequency (97%) was observed in 0.5 µM IBA, after 4 weeks of culture. The rooted plantlets achieved a remarkable 86% survivability rate, when transferred to ex vitro conditions during acclimatization. Analysis of photosynthetic parameters and their characteristics during the acclimatization phase revealed a gradual decline in photosynthetic attributes during initial weeks; however, a significant improvement was noted as the growth proceeded. SEM analysis revealed the ultra-morphological structural differences between in vivo and in vitro derived leaves of S. tetrasperma. Moreover, DPPH assay observed differential antioxidant activity of in vitro raised plantlets throughout the acclimatization period. The GC-MS analysis from leaf extracts of donor plants and in vitro derived plantlets has revealed a broad spectrum of phytochemical compounds with significant pharmacological properties. No polymorphism in the banding pattern was found when the genetic fidelity of the regenerated plants was evaluated using SCoT primers, indicating the clonal stability of micropropagated plants. This study is the first to explore the use of mT in regeneration of S. tetrasperma, showing its more effectiveness than BA and Kn.

Genetic diversity and population structure of Turkish European chestnut (Castanea sativa) genotypes assessed using start codon targeted polymorphism (SCoT) markers

Abstract The European chestnut (Castanea sativa) is an important nut crop that grows naturally in the Black Sea and Aegean regions of Türkiye. This study examined the genetic diversity and population structure of chestnut genotypes from prominent regions in Türkiye using Start Codon Targeted Polymorphism (SCoT) markers. A total of 44 Turkish chestnut genotypes from the Aegean, Marmara, and Black Sea regions, along with a control group of French variety, were analyzed. The SCoT primers underwent tests to select the most suitable ones, producing 8 selected amplified fragments, 65.34% of which were found to be polymorphic. The unweighted pair group method with arithmetic mean (UPGMA) and principal coordinate analysis (PCoA) analyses showed clear discrimination between two populations based on their origins, which was supported by the population structure analysis. The analysis of molecular variance (AMOVA) analysis revealed that 3% of the genetic variation was within populations and 97% was among individuals. The out-group (French variety) showed the furthest genetic similarity, and genetic similarity values decreased with increasing geographic distance. The SCoT primers successfully fingerprinted chestnut genotypes and could be used in future research to analyze the phylogeny of chestnuts using genomic DNA.

Genetic polymorphism amongst moth bean (Vigna aconitifolia (Jacq.) Marechal) accessions evaluated using various molecular markers

This study presents genetic level diversity of 50 moth bean legume accessions (Vigna aconotifolia (Jacq.) Marechal). Despite seeds being a rich source of soluble nutritional proteins, it still remains an underutilized crop commercially. To assess the molecular diversity amongst accessions, RAPD, ISSR, DAMD, SCoT and SSR were employed as PCR markers. Of the total 5064 polymorphic bands shown with these makers, DAMD1 and SCoT1 presented higher levels of polymorphism with 407 and 262 polymorphic bands respectively. These genotypes revealed logical similarity and variation under UPGMA clustering. The PCA analysis of IC-258167, IC-415167, and IC-402286 present three highest eigen values, hence seem fit to be considered as parents for moth bean breeding programme. The polymorphic bands as produced by SCoT and DAMD primers can serve unique genotypic identifiers. This study seems to present a maiden report in labelling genetic marker based polymorphism amongst the accessions of moth bean.

Evaluation of genetic diversity within different rabbit (Oryctolagus cuniculus) genotypes utilizing start codon targeted (SCoT) and inter-simple sequence repeat (ISSR) molecular markers

Abstract. This work aimed at studying the genetic diversity among different rabbit genotypes reared in Egypt by two different molecular markers (start codon targeted, SCoT, and inter-simple sequence repeat, ISSR) to improve breeding strategies. Five different groups of rabbits were used Gabali (Gab), New Zealand white (NZW), Californian (Cal), Rex, and Papillon (Pap). DNA was extracted and analyzed using SCoT and ISSR-PCR, and the obtained fragments were analyzed. Six SCoT primers scored 60 bands with 78.33 % polymorphism; primer SCoT 6 was the most polymorphic marker with 92.31 % polymorphism, while SCoT 5 was the lowest with 60 %. A dendrogram based on SCoT-PCR revealed that the Rex breed was the most genetically different. Seven ISSR primers gained 56 bands in total with 49.762 % polymorphism. ISSR 4 was the most polymorphic primer that detected 75 % of polymorphism, while ISSR 6 was not able to detect any polymorphism. It was suggested that the SCoT markers may be more effective than ISSR for differentiating and identifying the genetic variations within investigated breeds. Also, the usage of molecular markers of SCoT and ISSR may be more proper for calculating genetic diversity and common ancestry among tested rabbit breeds. Furthermore, evaluating genetic variability is important for enhancing existing breeds' adaptation to ecological alterations and crucial for preservation or breeding purposes.

Applicability of SCoT markers in unraveling genetic variation and population structure among sugar beet (Beta vulgaris L.) germplasm.

Sugar beet (Beta vulgaris L.) holds significant importance as a crop globally cultivated for sugar production. The genetic diversity present in sugar beet accessions plays a crucial role in crop improvement programs. During the present study, we collected 96 sugar beet accessions from different regions and extracted DNA from their leaves. Genomic DNA was amplified using SCoT primers, and the resulting fragments were separated by gel electrophoresis. The data were analyzed using various genetic diversity indices, and constructed a population STRUCTURE, applied the unweighted pair-group method with arithmetic mean (UPGMA), and conducted Principle Coordinate Analysis (PCoA). The results revealed a high level of genetic diversity among the sugar beet accessions, with 265 bands produced by the 10 SCoT primers used. The percentage of polymorphic bands was 97.60%, indicating substantial genetic variation. The study uncovered significant genetic variation, leading to higher values for overall gene diversity (0.21), genetic distance (0.517), number of effective alleles (1.36), Shannon's information index (0.33), and polymorphism information contents (0.239). The analysis of molecular variance suggested a considerable amount of genetic variation, with 89% existing within the population. Using STRUCTURE and UPGMA analysis, the sugar beet germplasm was divided into two major populations. Structure analysis partitioned the germplasm based on the origin and domestication history of sugar beet, resulting in neighboring countries clustering together. The utilization of SCoT markers unveiled a noteworthy degree of genetic variation within the sugar beet germplasm in this study. These findings can be used in future breeding programs with the objective of enhancing both sugar beet yield and quality.

Comparing genetic differentiation and variation using ISSR and SCoT among Juniper plant markers in Saudi Arabia.

Juniperus, a genus of fragrant evergreen trees in the Cupressaceae family, encompasses up to 67 distinct species distributed globally. Among these, Juniperus procera, also known as the East African pencil cedar or African Juniper, stands out for its notable medicinal properties. Due to the well-recognized therapeutic benefits of Juniper species, assessing genetic diversity is essential for various breeding initiatives. Thus, in this work, six ISSR and six SCoT primers were utilized to evaluate the genetic diversity between 23 Juniper genotypes collected from different locations in Saudi Arabia. 29 out of 103 and 26 out of 105 amplified bands, respectively, were found to be polymorphic markers using the ISSR and SCoT studies. With the help of 120 genotype-specific markers, including 60 for ISSR and 60 for SCoT, several varieties of Juniper were discovered. In addition, the polymorphism information content (PIC) was computed to assess the effectiveness of the markers. The findings of this study highlight the importance of conserving the genetic diversity of Juniperus procera, as it holds immense potential for developing new medicinal products. Additionally, the results provide valuable insights into the genetic structure of Juniper populations in western Saudi Arabia, which can inform future conservation and management efforts. However, all of the techniques utilized to profile the genotypes of Juniper can be regarded as useful techniques for long-term fingerprinting and diagnostic markers.

Optimization of PCR Analysis Based on Start Codon Targeted Markers (SCoT Markers) for Identification of Genetic Variation of Seaweed from Central Sulawesi

Seaweed is a fishery commodity that has high economic value because it contains carrageenan which can be used in the pharmaceutical, food, cosmetics, and industrial fields. Central Sulawesi Province is the second largest seaweed producer in Indonesia after South Sulawesi Province. The genetic diversity of seaweed in Central Sulawesi has not been studied much. Therefore, molecular-based characterization activities to identify the genetic variation of seaweed in the province are urgently needed. The purpose of the study was to optimize PCR techniques based on targeted start codon markers (SCoT markers) to identify the genetic diversity of seaweed accessions from Central Sulawesi Province. The results showed that the parameters for seaweed PCR amplification have been optimized. Of the fifteen SCoT primers, 10 of them can work to amplify seaweed DNA collected from several regions in Central Sulawesi. This is shown by the production of PCR result amplicons. PCR results also indicate polymorphism from the seaweed samples tested, although some seaweed samples have not been successfully amplified. The parameters of the PCR technique still need to be improved, especially the concentration of DNA prints, so that valid PCR results will be obtained for use in identifying seaweed genetic diversity.

Morphological, Biochemical, and Molecular Diversity Assessment of Egyptian Bottle Gourd Cultivars.

The genetic variability and relationships between ten bottle gourd cultivars were evaluated based on morphological, biochemical, and molecular parameters. The results displayed high variability among selected cultivars in terms of photosynthetic pigments, total free amino acids, total phenol content, isozymes pattern, and protein electrophoresis. Furthermore, differences in molecular markers were revealed by the SCoT technique. The peroxidase (POD) and polyphenyl oxidase (PPO) isozymes patterns did not detect significant differences in bands among cultivars. The protein patterns revealed seventeen bands ranging from 126 to 9 kDa and five polymorphic bands representing 29.41%. On the other hand, eight SCoT primers were used to evaluate the genetic variability and relationships between the ten Egyptian bottle gourd cultivars. The results of SCoT analysis detected 44 amplicons with 50% polymorphism. In addition, the results of the phylogenetic tree that is constructed based on the similarity coefficient revealed by SCoT analysis confirm the results of biochemical analysis indicating a genetic relationship between the most efficient bottle gourd cultivars (S1 and S2 cultivars). In addition, there is a genetic relationship among the less efficient bottle gourd cultivars (S4 and S5 cultivars). These results could be beneficial to distinguish among bottle gourd cultivars in the plant breeding programs.

Effectiveness of DNA barcoding, SCOT markers and phytochemical characterization in biodiversity assessment of some Zea mays hybrids

Because of the rapid fluctuation in world's climate, politics, and economy, maize outputs must be increased to meet food security demands. Due to the importance of the genetic materials under study as a source of quality traits and genetic variability, this study was designed to examine the effectiveness and strength of assessing biodiversity in some maize hybrids using barcodes and molecular SCoTs markers in addition to phytochemical characterization. Using matK and rbcL sequences, pairwise distances were obtained and assessed. The hybrids under study were distributed into different groups based on the matK and rbcL sequences. The 12 SCoT primers amplified 155 amplicons with an average of 12.92 bands per primer, of which 63 monomorphic bands (MB) (40.65 %) and 92 polymorphic band (PB) (59.35 %). Additionally, the number of unique bands was six bands. The SCoT-6 primer produced three bands individually, while the primers SCoT-2, SCoT-3, SCoT-4, and SCoT-7 produced one band each. Hybrids S.C. 10, S.C. 128, S.C. 130, S.C. 131, S.C. 132 and TWC. 324 were more influenced by some primary constituents and bioactive secondary compounds such as moisture content (%), crude fiber, total carbohydrates, total sugars, soluble carbohydrates, non-reducing sugars, phenolics compounds, and tannins. The hybrids S.C. 168, S.C. 176, S.C. 178, TWC. 360, TWC. 368 and TWC. 321 were more influenced by the SCoT molecular markers. The results of genetic variation between the studied hybrids could be considered a potentially source of genetic information in developing future breeding programs.

Evaluation of genetic diversity using iPBS-SCoT marker methods in native hawthorn genetic resources and species ıdentification by using DNA barcoding method.

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 Çoruh Valley, Türkiye 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 primers 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.

The Potential Genetic Effect for Yield and Foliar Disease Resistance in Faba Bean (Vicia faba L.) Assessed via Morphological and SCoT Markers.

Faba bean is considered one of the most prominent grain legumes, with high protein content for human food consumption and livestock feed. The present study evaluated the nature of gene action and determined the genetic diversity among different populations of three crosses for resistance to foliar diseases at the molecular level. Analysis of variance exposed significant differences among the generations for all measured traits. Both dominance and additive gene effects were essential, but dominance genes, for the most part, exhibited greater effects than additive ones. This indicates an essential role for dominant genes alongside the additives one in inheriting such traits. The third cross (Marina × Giza 40) gave desired significant and positive (additive × additive) values for the number of pods/plant, seeds/plant, and seed yield/plant, in addition to desirable negative values for chocolate spot and rust characteristics. Furthermore, assessing the lines under study using seven SCoT primers disclosed three bands with recorded molecular weights of 260, 207, and 178 bp, generated by SCoT-1, SCoT-4, and SCoT-7 primers, respectively. These bands exist in the resistant parent (Marina), which could be attributed to the high-disease-resistance phenotypes, and they are absent in the sensitive parent (Giza 40) and other putative sensitive lines. Based on the molecular profiles and the genetic similarity between parents and the selected lines, the highest similarity value (0.91) was detected between Marina genotype and BC1, revealing a high foliar disease resistance. Meanwhile, Giza 40 (susceptible to foliar diseases) exhibited the maximum value (0.93) with F2. Additionally, cluster analysis based on genetic relationships was performed, and a high level of correlation between the results of PCR-based SCoT analysis and the foliar disease reactions was observed in the field. Consequently, this study concluded that SCoT markers created reliable banding profiles for evaluating genetic polymorphism among faba bean lines, which could be a foundation for developing an efficient breeding program.

True-to-typeness and phytomedicinal potential in somatic embryo-derived plants of Crinum malabaricum (Amaryllidaceae): A medicinally important source of pharmaceutical biomolecules

Crinum malabaricum, Lekhak & S.R. Yadav (Amaryllidaceae) is a critically endangered bulbous plant endemic to India. Establishing an efficient plant regeneration protocol is necessary for its conservation and large-scale propagation. In the present study, regeneration was achieved through somatic embryogenesis from callus produced on MS media supplemented with various different concentrations of 2,4-D alone and in combination with N6-benzyl-adenine (BA). Different advanced stages of embryo development (globular, torpedo and cotyledonary) and maturation were obtained on MS medium, with combinations of picloram and thidiazuron (TDZ). A high rate of somatic embryos (55.89 ± 0.60) was obtained after eight weeks. SEM examinations showed the occurrence of cell clusters (embryogenic) which converted to somatic embryos. Well-developed cotyledon embryos were successfully germinated on PGRs free full and half strength MS medium, however, 94.03% of somatic embryos germinated on half-strength MS medium supplemented with 1.0 mg L−1 gibberellic acid. The true-to-type genetic conformity of regenerated plants was examined by SCoT, ISSR and RAPD primers based molecular analysis. The amplification bands produced by SCoT, ISSR and RAPD primers applied were monomorphic across all the regenerated plants. This confirmed their genetic homogeneity compared to the mother plant and also demonstrated the reliability of our somatic embryogenesis system for C. malabaricum. A rapid method for phytochemical analysis based on LC-ESI/MS exhibited better separation and analysis of galanthamine and lycorine from methanolic extracts of in vitro raised plants derived from somatic embryogenesis. The protocol developed should be helpful in reintroduction, restoration and ex situ conservation of this valuable plant in the natural condition as well as in the pharmaceutical sectors.

Intergeneric hybridization of two endangered orchids, Vanda stangeana and Phalaenopsis hygrochila, and molecular confirmation of hybridity using SSR and SCoT markers

Vanda stangeana Rchb. f. is an endemic species of Manipur, India. Its flowers are greenish-yellow, 5 cm in size with a pink mid-lobe, tessellated in chestnut brown color and tough texture with a long shelf life. Phalaenopsis hygrochila J.M.H. Shaw is a large-sized, hot to cool growing, epiphytic orchid with sturdy, 20 to 40 cm, more or less flattened stem with elliptic-oblong leaves. The flowers are fragrant, pale greenish yellow, more or less dotted with brown or brown-red, with a whitish or white base, waxy, long-lasting, up to 6 cm in diameter, column white, labellum slightly purplish purple, paler towards the margins. Both plants are included under CITES' Appendix II. The current study concentrates on a hybridization program involving two vandaceous orchid genera, namely Vanda stangeana and Phalaenopsis hygrochila to investigate their cross-ability and molecular confirmation of the synthesized hybrid using SSR and SCoT marker system. MS medium at half-strength enriched with 0.5 mg/L NAA + 0.5 mg/L Kn was the optimal medium for the development of the synthesized hybrid seedlings with the longest shoot (2.6 cm), the maximum number of leaves (4.5), leaf length (2.5 cm) and highest number of root (4.4) after 150 days of culture on the medium. In total, 65 amplified bands were revealed by seven SCoT primers (SCoT4, SCoT6, SCoT9, SCoT10, SCoT34, SCoT35 and SCoT36), of which 38 (58.46%) were polymorphic with the percentage ranging from 22.22% to 85.75%, thereby convincingly demonstrated the hybridity. Out of 8 pairs of SSR primers used, 3 primers (C106, MOK26 and C268) showed complementary banding patterns. Our study demonstrated that an intergeneric hybrid can be successfully produced by the hybridization of vandaceous orchids belonging to two different genera.

Exploring the morphological and genetic diversity of Egyptian basil landraces (Ocimum sp.) for future breeding strategies

BackgroundBasil (Ocimum sp.) exhibits significant morphological and genetic diversity. This variation provides an opportunity to identify novel traits that can be used in breeding programs to improve the productivity, quality, and disease resistance of basil plants. The integration of morphological and genetic data for basil varieties can improve our understanding of biodiversity, conservation, and breeding programs.ResultsIn 2019, we conducted a comprehensive investigation on 25 basil landraces collected from 12 Egyptian governorates to explore their morphological and genetic diversity. Morphological characterization revealed variability in plant vigor, flower color, cyme shape, and leaf shape, indicating distinct growth patterns and potential for specific applications. Productivity and biochemical evaluation demonstrated significant variation in yield and oil distillation, identifying top-performing landraces L5, L11, and L24 that can be used in breeding programs to optimize performance and reduce environmental impact. The genetic diversity of the landraces was investigated using 11 SCoT primers revealed high levels of genetic diversity among landraces, with a wide range of polymorphism variation. The phylogenetic analysis identified two distinct clades, providing valuable insights into genetic diversity and relationships between different landraces. An interesting observation was made in the study, whereby L6 and L13, collected from Fayoum and Port-Said, respectively, were found to be the closest landraces. Following closely were L7 and L14, which were also collected from the same governorates. These findings have significant implications for the conservation of these landraces, as they may have evolved from similar species.ConclusionsThis study sheds light on the genetic diversity and relationships among 25 Egyptian basil landraces. Using SCoT markers, a high level of polymorphism was detected, indicating significant variation in the genetic makeup of the landraces. The study also revealed interesting observations regarding the relationships among the landraces, with some landraces appearing to have evolved from similar species. The phylogenetic analysis provided insights into the genetic relationships among the landraces, which can guide conservation efforts and breeding programs. Overall, this study provides valuable information for researchers, breeders, and farmers involved in the conservation, breeding, and utilization of basil genetic resources in Egypt.

Indirect shoot regeneration from root explants, assessment of clonal fidelity of regenerated plants using SCoT primers and antioxidant analysis in Thottea siliquosa (Lamk.) Ding Hou

Indirect shoot regeneration from root explants, assessment of clonal fidelity of regenerated plants using SCoT primers and antioxidant analysis in Thottea siliquosa (Lamk.) Ding Hou

Deciphering the level of genetic diversity in some aegilops species using CAAT box-derived polymorphism (CBDP) and start codon target polymorphism (SCoT) markers.

Maintaining genetic diversity is of the most essential principle for a long-term conservation of plant genetic resources and could play a crucial role in their management. The genus Aegilops is one important member of wheat germplasm, and there are evidences that novel genes of this genus' species can be studied/utilized as ideal sources for the wheat cultivar improvement. The objective of this study was to dissect the genetic diversity and population structure among a set of Iranian Aegilops using two gene-based molecular markers. This study investigated the level of genetic diversity among 157 Aegilops accessions consisting of Ae. tauschii Coss. (DD genome), Ae. crassa Boiss. (DDMM genome), and Ae. cylindrica Host. (CCDD genome) belonging to NPGBI using two sets of CBDP and SCoT markers. The SCoT and CBDP primers yielded 171 and 174 fragments, out of which 145 (90.23%) and 167 (97.66%) fragments were polymorphic, respectively. The average of polymorphism information content (PIC)/ marker index (MI)/resolving power (Rp) for SCoT and CBDP markers were 0.32/3.59/16.03 and 0.29/3.01/16.26, respectively. Results of AMOVA revealed the genetic variability within species was greater than the variation observed among them (SCoT: 88% vs. 12%; CBDP: 72% vs. 28%; SCoT + CBDP: 80% vs. 20%). Based on the information obtained from both markers, the higher level of genetic diversity was found in Ae. tauschii as compared to other species. The grouping patterns obtained by Neighbor-joining algorithms, principal coordinate analysis (PCoA), and Bayesian-model-based structure were consistent with each other and resulted in grouping all studied accessions according to their genomic constitutions. The results of this study revealed a high level of genetic diversity among Iranian Aegilops germplasm. Moreover, SCoT and CBDP marker systems were efficient in deciphering DNA polymorphism and classification of Aegilops germplasm.

Changes in morphological traits, anatomical and molecular alterations caused by gamma-rays and zinc oxide nanoparticles in spinach (Spinacia oleracea L.) plant

Spinach seeds were irradiated with gamma-rays after that soaked in zinc oxide nanoparticles (ZnO–NPs) at 0.0, 50, 100 and 200 ppm for twenty-four hours at room temperature. Vegetative plant growth, photosynthetic pigments, and proline contents were investigated. Also, anatomical studies and the polymorphism by the SCoT technique were conducted. The present results revealed that the germination percentage was at the maximum values for the treatment of 100 ppm ZnO–NPs (92%), followed by 100 ppm ZnO–NPs + 60 Gy (90%). The application of ZnO–NPs resulted in an enhancement in the plant length. The maximum of chlorophylls and carotenoids content was recorded in the treatment, 100 ppm ZnO–NPs + 60 Gy. Meanwhile, the irradiation dose level (60 Gy) with all ZnO–NPs treatments increased proline content and reached its maximum increase to 1.069 mg/g FW for the treatment 60 Gy combined with 200 ppm ZnO–NPs. Also, the anatomical studies declared that there were variations between the treatments; un-irradiated and irradiated combined with ZnO–NPs plants which reveal that the leave epidermal tissue increased with 200 ppm ZnO–NPs in both the upper and lower epidermis. While irradiated plants with 60 Gy combined with 100 ppm ZnO–NPs gave more thickness of upper epidermis. As well as SCoT molecular marker technique effectively induced molecular alterations between the treatments. Where, SCoT primers targeted many new and missing amplicons that are expected to be associated with the lowly and highly expressed genes with 18.2 and 81.8%, respectively. Also, showed that the soaking in ZnO-NPs was helped for reducing molecular alteration rate, both spontaneous and induced by gamma irradiation. This nominates ZnO–NPs as potential nano-protective agents that can reduce irradiation-induced genetic damage.

Investigation of molecular variability in some Aegilops species using Start Codon Targeted Polymorphism (SCoT) and CAAT-Box Derived Polymorphism (CBDP) markers

Among wild relatives of wheat, Aegilops species are ideal genetic resources for the discovery of new characteristics such as resistance to environmental stresses and even grain quality for wheat improvement. Hence, knowledge of the population structure and genetic diversity of this germplasm is very important for their conservation and further utilization. In the present study, 80 accessions of the Aegilops including Ae. tauschii, Ae. cylindrica and Ae. triuncialis were investigated for genetic diversity using SCoT and CBDP markers. Eight SCOT and twelve CBDP primers amplified a total of 84 and 94 fragments with a mean of 10.50 and 7.83 fragments per primer, respectively. Resolving power (Rp) for SCoT and CBDP primers varied between 6.04 and 11.65, and 13.08 and 28.02, with the polymorphic information content (PIC) from 0.40 to 0.49 and 0.35 to 0.48, respectively. The results of analysis of molecular variance (AMOVA) indicated that the highest proportion of genetic variance referred to between species. SCoT primers indicated high values for all informativeness parameter (except resolving power) than CBDP primers across all tested accessions. However, CBDP primers indicated higher values of the genetic parameters than using SCoT primers. As a result, the maximum values for genetic parameters such as number of effective alleles (Ne), Nei?s gene diversity (H) and Shannon?s information index (I) were detected in Ae. cylindrica and Ae. triuncialis using SCoT and CBDP markers, respectively. Cluster analysis based on those molecular system grouped all accessions into three main clusters. The grouping pattern observed by CBDP primers indicated more clear phylogenetic relationship among some Aegilops species, so that PCoA?s results confirmed the grouping pattern. In conclusion, it was observed that SCoT and CBDP displayed good efficiency in depicting polymorphism among the tested accessions, however, CBDP markers provided a clear grouping pattern of evaluated accessions. Hence, the use of CBDP markers in determining population structure and estimating genetic diversity in other plant species is recommended.

Biotechnological studies on Arthrospira platensis biomass cultivated in enriched culture with chelated leather waste and chelated glycinate

The current work aims to evaluate the growth rate, molecular patterns and biological activities of Arthrospira platensis cultivated in culture enriched with different chelated minerals (Fe, Zn, Cu, Mn, and Mg) of leather protein hydrolysates (LPHs) and glycine in addition to determine the phytochemical contents of Arthrospira platensis cultivated in modified Zarrouk medium. Also, the work study the protein profile with molecular variations (using SDS-PAGE, Comet assay, and SCOT-PCR) in collected biomasses when compared with control (cyanobacteria cultivated in Zarrouk medium). Superoxide dismutase (SOD), glutathione peroxidase (GPX), and mineral content of each treatment were also determined. The obtained Results showed that a significant increment of growth rate of A. platensis in glycinate treatments was observed especially with Cu-glycinate more than control medium. Additionally, T11 has high O.D. than other chelated minerals but control medium had the highest O.D. than LW-chelated minerals. All A. platensis treated with the glycine-chelates possessed both conserved and additional proteins in their SDS-PAGE banding patterns. Moreover, A. platensis treated by chelated minerals with leather waste hydrolysate induced high discrimination than their analogues chelated with glycine. Six ScoT primers were applied and produced 323 amplicons with an average of 68% polymorphism/primer. SDS and Scot dendrogram revealed that among all treatments, control and Mg-glycine-chelated treatment were closely related. All A. platensis samples treated by chelated minerals with leather waste hydrolysate possessed more polymorphic bands than monomorphic ones (either as proteins or DNA), despite that no significant DNA damage (as % tDNA) was detected on A. platensis treated with either chelated treatment.

Genetic diversity using biochemical, physiological, karyological and molecular markers of Sesamum indicum L.

The genetic diversity and the relationships among sesame cultivars were investigated using physiological and cyto/molecular analysis. To our information, no studies have yet been conducted on the genetic evaluation of sesame genotypes based on cyto/molecular analysis in Saudi Arabia. This study showed that genotype Bah-312 had the highest values from physiological and biochemical traits (plant height, harvest index, total plant dry matter, seed yield, oil content, and fatty acids content). Using 20 ISSR and 25 SCoT primers, the studied genotypes amplified 233 and 275 alleles, while the average polymorphism percentage (P%) was 65.32% (ISSR) and 77.8% (SCoT) across all the studied genotypes, respectively. To assess the markers efficiency analysis the polymorphism information contents (PIC), Marker Index (MI), Effective Multiplex Ratio (EMR), Resolving Power (Rp) were estimated. In general, primers (ISSR 2 & SCoT 21) and (ISSR 4 & SCoT 3) revealed the highest and lowest values for P %, PIC, MI, and EMR%. Furthermore, 188 positive and negative unique bands were detected, out of which ISSR generated 84, while 104 were amplified by SCoT analysis. In this regard, genotype Bah-312 generated 41 unique amplicons, and Jiz-511 genotype 23 unique amplicons. In the same context, the population genetics parameters, number of different alleles (Na), number of effective alleles (Ne), Shannon’s index (I), expected heterozygosity (He), and Unbiased Expected Heterozygosity (uHe), were calculated. ISSR marker showed the highest values for all the estimated parameters. In this regard, genotype Bah-312 exhibited the highest values (1.35, 1.37, 0.31, 0.21, 0.29) & (1.31, 1.35, 0.30, 0.20, 0.27) while, genotype Ahs-670 revealed the least values (1.29, 1.31, 0.26, 0.16, 0.23) &(1.14, 1.26, 0.22, 0.15, 0.20) for ISSR and SCoT markers respectively. For cytological data, according to the highest asymmetry index (AsK%) and lowest total form percentage (TF%) values, genotype Ahs-670 was the most advanced cultivar, and genotype Bah-312 was the most primitive one. According to the degree of asymmetry of karyotype (A) and intrachromosomal asymmetry index (A1), sesame genotype Ahs-670 was the most asymmetrical, and Bah-312 was the most symmetrical genotype. This study gives some helpful information about the genetic diversity of six sesame landraces. The variation harbored by these landraces could be used in sesame breeding programs.