Search Results (72)

Indian senna (Senna alexandrina Mill.), a perennial medicinal species belonging to the family Fabaceae, holds significant therapeutic and commercial importance owing to its rich content of sennosides and rhein derivatives, which confer well-established laxative properties. Its high market demand, however, renders the species vulnerable to deliberate or inadvertent adulteration. While previous investigations have utilized functional marker systems such as SCoT (Start Codon Targeted Polymorphism)- and CBDP (CAAT Box Derived Polymorphism)-derived SCAR (Sequence Characterised Amplified Region) markers for genetic characterization, the present study is the first to report the development of sequence-specific RAPD- and ISSR-based SCAR markers consolidated into a single-tube multiplex PCR assay. Genomic DNA isolated from young leaves of S. alexandrina and its commonly encountered adulterant species was amplified using RAPD primer OPI-02 and ISSR primer UBC-835. Polymorphic amplicons were cloned, sequenced, and employed for the design of SCAR primers, which were rigorously validated for specificity. Species-specific SCAR markers were successfully integrated into a single multiplex reaction, enabling precise and unequivocal identification of S. alexandrina, Cassia fistula and Senna sophera. The multiplex amplification profiles were entirely consistent with corresponding uniplex assays, endorsing the method’s robustness and reproducibility. This streamlined, one-tube multiplex SCAR-PCR system represents a significant advancement toward reliable, high-throughput molecular authentication of Indian senna and its closely related medicinal plant species (adulterants). Full article

Platonia insignis Mart. (Clusiaceae) is a native fruit tree of great ecological and socioeconomic importance in the Brazilian Amazon and Cerrado. However, habitat loss is threatening its genetic variability. We investigated whether habitat fragmentation across the Amazon, Cerrado, and transition zones shapes the genetic diversity and population structure of five natural populations of P. insignis, using ISSR markers. Leaf samples from 13–15 individuals per population were collected, and DNA was extracted using the CTAB protocol. Twelve ISSR primers amplified 149 loci, used to estimate genetic parameters. AMOVA showed that 73.58% of genetic variation occurred within populations and 26.41% among populations (F_ST = 0.261). Amazonian populations exhibited the highest genetic diversity, while transition zone populations had the lowest values. The Cerrado population was genetically distinct and maintained moderate intrapopulation diversity. Bayesian clustering, PCoA, and UPGMA revealed three genetic groups corresponding to the sampled regions. Transitional populations showed high genetic admixture, indicating their role as potential corridors for gene flow. Our results highlight the need to preserve genetically diverse Amazonian populations, safeguard the Cerrado population as an evolutionarily significant unit, and maintain transitional populations to promote landscape connectivity. The study provides a genetic baseline to support conservation and management of P. insignis germplasm resources. Full article

Sugar beet (Beta vulgaris L.) is an economically important crop that accounts for approximately 20% of global sugar production. The success of future breeding programs depends on the effective utilization of existing genetic resources. The aim of this study was to assess the genetic diversity and population structure of 192 sugar beet (Beta vulgaris L.) genotypes, including commercial cultivars and accessions obtained from the USDA gene bank, using SCoT and ISSR molecular markers, and to identify potential genetic resources for sugar beet breeding programs. In this study, a total of 192 sugar beet genotypes, including 187 accessions from the USDA (U.S. Department of Agriculture) gene bank and 5 commercial cultivars, were evaluated for genetic diversity using Start Codon Targeted (SCoT) and Inter Simple Sequence Repeat (ISSR) markers. A total of 68 scorable bands were obtained from five SCoT and three ISSR primers, and all bands were found to be polymorphic (100% polymorphism). Parameters such as polymorphic information content (PIC), Nei’s genetic diversity, and Shannon’s index indicated a high level of variation within the gene pool, with SCoT markers being more informative than ISSR markers. Dendrogram analyses based on Nei’s genetic distance revealed that the populations were separated into two main groups, while the sub-clusterings contained broad genetic variation. STRUCTURE analysis identified four (K = 4) populations for the SCoT data and three (K = 3) populations for the ISSR data; the inclusion of a high number of individuals in the admixture population indicated extensive gene flow. Principal component analysis (PCA) revealed both homogeneous groups and differentiated genotypes contributing to within-population diversity. The results demonstrate that the combined use of SCoT and ISSR markers provides powerful and complementary tools for assessing genetic diversity in sugar beet. The findings provide a solid scientific basis for the development of new, high-yielding and high-quality sugar beet cultivars as well as for the conservation of existing genetic resources. Molecular data constitute an important reference for guiding sugar beet breeding programs and for the effective utilization of genetic resources. Full article

Lavender has been cultivated in Bulgaria for over a century. The high essential oil content and quality of Bulgarian lavender varieties have established the country as a leading global producer. Studies into the crop’s genetic diversity are essential for selecting varieties best suited to specific environmental conditions, maximizing resilience and yield. Therefore, identifying appropriate genetic markers to monitor lavender diversity is a key prerequisite for developing effective crop selection strategies, particularly in response to the challenges posed by global climate change. In this study, we evaluate the versatility of markers for assessing genetic diversity of lavender genotypes. A total of 96, 97 and 96 bands were recorded using the 13 Start Codon Targeted Polymorphism (SCoT), 13 Inter-Simple Sequence Repeat (ISSR) and 14 Cis-Element Aligned Polymorphism (CEAP) primers, respectively. All amplification programs used were successful in the studied genotypes. Additionally, four informative primers of each marker system were applied for assessment of the within-field genetic variability in two lavender plantations from Bulgaria. This is the first report on the combined use and comparison of CEAP, SCoT and ISSR primers in lavender genotypes in Bulgaria. Full article

Rivomarginella morrisoni (Gastropoda: Marginellidae) is a narrowly distributed freshwater snail inhabiting drainage basins of central and southern Thailand. To clarify patterns of genetic differentiation across its range, 45 individuals from 11 sites across eight river basins were analyzed using two dominant molecular markers: sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeats (ISSR). SRAP primers produced higher polymorphic information content (PIC) values than ISSR primers (0.35 vs. 0.27). Analysis of molecular variance (AMOVA) revealed strong population structure, with 80.29% of the genetic variation occurring among populations and 19.71% within populations Population differentiation statistic (PhiPT) = 0.803, p < 0.001). Unweighted Pair Group Method with Arithmetic mean (UPGMA) and principal coordinate analysis (PCoA) consistently separated central and southern populations, and STRUCTURE supported K = 2 as the most likely number of clusters. Similarly, principal component analysis (PCA) of morphological traits also distinguished specimens into two groups corresponding to these geographic regions, confirming region-specific divergence. Overall, the genetic and morphological patterns indicate restricted gene flow among basins and a clear separation between central and southern lineages of R. morrisoni. This study provides the first molecular evidence of population structure in this species and offers important baseline information for future taxonomic, ecological, and conservation research on freshwater marginellid snails. Full article

Major historic olive tree cultivars around the Mediterranean originate from the Jordan area and possess a proven abiotic stress tolerance; however, they were unexplored from the diversity perspective. Therefore, historic olive tree accessions from three northern regions—Irbid (i), Jerash (J), and Ajloun (A)—were analyzed using DNA molecular markers to identify and study their genetic relationships and genetic structure. DNA molecular markers of inter-simple sequence repeats (ISSR) were used. A total of 3150 data entries (859 present and 2291 absent) were generated with fragment sizes ranging from 350 to 2000 bp. Data entries were evaluated with UPGMA and population genetic structure analysis. The results showed that similarity among the investigated sixty-three accessions ranged from 9% between J14 and i20 up to 100% between ‘J11’ and ‘J12’ and between A8 and A9. The discriminating power values for ISSR_807, ISSR_810, and ISSR_825 were 0.70, 0.61, and 0.83, respectively. A generated dendrogram showed ten major clades, while the genetic structure could resolve four unique genetic pools: one for Irbid, one for Jerash, and two for Ajloun. In addition, analysis of 19 phenotypic parameters covering leaf, fruit, stone, and flesh was able to confirm the molecular data. Phenotypic and ISSR data were analyzed using PCA, cluster, and Mantel tests. ISSR markers showed clear genetic differentiation among groups, whereas phenotypic traits displayed lower variation but a significant correlation with molecular diversity. Promising accessions with either pure or admixture genetic makeup were identified. The resolved genetic structure of the investigated historic olive accessions would open new frontiers for olive breeding and utilization, helping to overcome current production challenges and climate change limitations. Full article

The Chinese holly (Ilex chinensis Sims.), an evergreen tree species native to China, is distributed mainly in regions south of the Qinling Mountains and Huai River. This research aimed to characterize the molecular profiles and genetic relationships of 40 Chinese holly genotypes via inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) polymorphism markers. Genetic diversity analysis revealed that the ISSR markers detected 111 polymorphic bands from 13 primers, with a polymorphism rate of 88.10%. The analysis generated parameters such as the observed allele number (Na = 1.876), effective allele number (Ne = 1.461), Shannon’s information index (I = 0.271), and expected heterozygosity (H = 0.411). In comparison, the SCoT markers produced 65 polymorphic bands from the 6 primers, resulting in a 100% polymorphism rate, with Na = 2.000, Ne = 1.695, I = 0.393, and H = 0.575. Cluster analysis classified the 40 genotypes into two main clusters with genetic similarity coefficients of 0.69 (ISSR) and 0.55 (SCoT). The ISSR markers presented the greatest similarity between the ZSS and ZLS genotypes, whereas the ZZDH and ZWW genotypes presented lower similarity. Conversely, the SCoT markers identified ZZP and ZJDS as the most similar, with ZLJ and ZHX showing less similarity. These results provide a theoretical basis for hybrid breeding, germplasm innovation, and conservation strategies of Chinese holly in China. Full article

The nopal cactus, a plant from the Cactaceae family, holds significant economic and nutritional value for Mexico. This study aimed to enhance the genetic diversity and morphological traits of Opuntia velutina, a species cultivated as a vegetable nopal. A total of 1050 in vitro O. velutina explants were exposed to 15 different doses of gamma radiation from ⁶⁰Co gamma, ranging from 5 to 125 Gy. The lethal dose was above 50 Gy, with an LD₅₀ of 22.8 Gy for stimulating in vitro shoot growth. Shoots derived from doses between 5 and 50 Gy were subjected to in vitro shoot proliferation across four consecutive generations to stabilize morphological traits. Cluster analysis categorized the 178 irradiated shoots into 13 distinct morphological groups (CG1–CG13). Twenty-seven shoots exhibiting significant morphological improvements, such as a 50–100% increase in cladode length, up to a six-fold increase in shoot number, and up to a seven-fold increase in root number, were selected for molecular analysis of genetic diversity. Six primers were used with the Inter Simple Sequence Repeat (ISSR) molecular markers to examine genetic uniformity, yielding 54.5% polymorphic bands, indicating a high level of genetic variation. Both a UPGMA dendrogram and STRUCTURE-based Bayesian analysis confirmed the genetic divergence among the selected mutant lines. Overall, gamma irradiation effectively enhanced both phenotypic and genotypic diversity in O. velutina. This study corroborates that in vitro mutagenesis through gamma radiation is a viable strategy for generating novel genotypes with breeding potential within the Opuntia genus. Full article

Melina (Gmelina arborea Roxb.) is a tree native to Asia, whose timber is not utilized in that region for a variety of reasons. However, the tree’s fast growth and extensive range of applications have increased its acceptance in other world’regions. G. arborea was introduced to Mexico in 1971, and it is currently the fifth most utilized forest species in commercial forest plantations (CFPs). However, its genetic diversity has not been evaluated in Mexico. The objective of this research was to investigate the genetic variability of Melina in Mexico using molecular markers. This investigation was undertaken to acquire valuable insights for the implementation of effective improvement strategies. A total of 85 Melina samples were collected from various locations in southeastern Mexico between 2017 and 2022. Genetic fingerprints were obtained using ten simple primer amplification reactions (SPARs): five Directed Amplification of Minisatellite DNA regions (DAMD), and five Inter-Simple Sequence Repeats (ISSRs). The polymorphic information content (PIC) was 0.940 and 0.950 for the DAMD and ISSR, respectively, and the similarity coefficients ranged from 0.12 to 0.88, indicating a high degree of polymorphism in the species under investigation. This is the first attempt to ascertain the genetic variability of Gmelina arborea in Mexico. Full article

In nature, orchid seed germination is extremely low, making in vitro asymbiotic seed germination essential for the propagation and conservation of endangered Vanda coerulea. This study optimized a micropropagation protocol and evaluated the genetic homogeneity of regenerated orchids. The synergistic effect of kinetin (KN) with auxins in the Mitra (M) medium best supported protocorm formation and seedling development. The highest shoot multiplication (5.62 ± 0.09) was achieved with 1.2 mg L⁻¹ KN and 0.6 mg L⁻¹ IBA (indole-3-butyric acid) in the medium. Enhanced leaf production (4.81 ± 0.37) was observed when 3.2 mg L⁻¹ KN was combined with 1.8 mg L⁻¹ IAA (indole-3-acetic acid), while root development was superior when 3.2 mg L⁻¹ KN together with 2.4 mg L⁻¹ IAA was incorporated in the medium. Anatomical sections confirmed well-developed leaf and root structures. Genetic fidelity assessment using random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), inter-primer binding site (iPBS), and start codon targeted (SCoT) markers revealed 97.17% monomorphism (240/247 bands) and low Nei’s genetic distances (0.000–0.039), indicating high similarity among the regenerants. Dendrogram clustering was supported by a high cophenetic correlation coefficient (CCC = 0.806) and strong resolution in Principal Coordinate Analysis (PCoA) (44.03% and 67.36% variation on the first two axes). The Mantel test revealed a significant correlation between both ISSR and SCoT markers with the pooled marker data. Flow cytometry confirmed the genome stability among the in vitro-propagated orchids, with consistently low CV (FL2-A) values (4.37–4.94%). This study demonstrated the establishment of a reliable in vitro protocol for rapidly propagating genetically identical V. coerulea via asymbiotic seed germination. Full article

Globally, people are cultivating finger millet, an important cereal, to improve food availability and health benefits for humans. However, the biotechnological research on this millet is limited and insufficient in this field. The primary focus of this study is to optimize an efficient regenerated protocol for initiating further plant transformation studies, using the shoot apex as an explant and various growth regulators. For example, three cytokinins (BAP, TDZ, and Kin) at different concentrations were used to induce multiple shoots of finger millet. Among these, TDZ (4.5 µM) provided the maximum number (17.3) of shoots as compared to BAP and Kin. IBA (2.46 µM), along with MS medium, was used for the induction of roots, where 5.6 roots were produced in an individual shoot and the length of the root was longer with a size of 8.2 cm after two weeks of incubation. The clonal fidelity of the in vitro regenerated plantlets of finger millet was confirmed by ISSR primers. Overall, the present work developed a robust and reliable procedure for the establishment of efficient and reproducible regeneration through the shoot apex that will be useful for the genetic improvement of this crop. The genetic enhancement of these millets as well as the successful creation of transgenic plant varieties modified for resistance to biotic and abiotic challenges in the near future would be aided by this study. Full article

Türkiye has a unique position globally as it connects the Asian and European continents. In this study, the genetic diversity of a total of 27 mulberry (Morus alba) genotypes collected from northeastern Türkiye was evaluated in terms of morphological, biochemical, and molecular characteristics. In the fruits of the mulberry genotypes, the fruit weight (FW) ranged from 2.00 to 5.13 g, fruit width (Fw) ranged from 12.64 to 16.93 mm, and fruit length (FL) ranged from 20.89 to 32.34 mm. Total soluble solids (TSS) ranged from 16.56% to 26.00%, fruit pH ranged from 3.87 to 5.10, total phenolic content (TPC) ranged from 72.69 to 171.34 mg GAE/100 g, total flavonoid content (TFC) ranged from 75.55 to 86.85 mg QE 100/g, and antioxidant activity (AA) ranged from 42.35% to 74.93%. According to principal component analysis (PCA), the first three principal components (eigenvalue ≥ 1.00) explain 54.05% of the total variation. In general, the fruit and leaf characteristics were more effective in the first two principal components, while the biochemical traits were more effective in the third principal component. As a result of molecular analysis using 13 inter-simple sequence repeat (ISSR) primers, 94.23% of the total of 76 bands was polymorphic. Molecular variance analysis (AMOVA) showed that within the population, genetic variation was 79%, and among the populations, the variation was 21%. The similarities of the genotypes in the UPGMA dendrogram created according to the results of the molecular analysis varied between 0.74 and 0.98, and the genotypes were divided into two main groups. This study guides breeders in future studies by evaluating mulberry genotypes from morphological, biochemical, and molecular perspectives and explaining the wide variation among genotypes. Full article

In this study, we present the first-ever comparison of the effectiveness of SRAP and ISSR markers on three Allium species. In addition, to visualize the results of each dataset in a simpler way, the Fruchterman–Reingold algorithm was used to generate a link graph and neighbor-joining methods were used to obtain a phylogenetic tree. The genetic similarity matrices were compared using the Mantel test. Primers generated 59 ISSR and 72 SRAP fragments. There was no statistically significant difference between the polymorphism information content of the marker sets. In terms of the effective multiplex ratio, SRAP markers were higher than ISSR markers, with values of 6.700 for garlic, 6.400 for onion, and 5.800 for leek (3.490, 4.316, and 2.573, respectively). Similarly, the marker index was calculated as 2.820, 3.056, and 2.505 for SRAP and 1.903, 1.523, and 1.050 for ISSR in onion, garlic, and leek species, respectively. The highest value regarding cophenetic correlation coefficients was obtained from the Jaccard method. According to the neighbor-joining method, the tree drawn using SRAP and ISSR data together shows a more distinct hierarchical structure of genotypes. The results obtained proved that SRAPs have higher values in terms of sign efficiency criteria, but they are not sufficient for the homogeneous grouping of different Allium species. Full article

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc), is recognized as one of the most devastating diseases affecting banana cultivation worldwide. In India, Foc extensively affects Malbhog banana (AAB genomic group) production. In this study, we isolated 25 Foc isolates from wilt-affected Malbhog plantations inIndia. A pathogenicity test confirmed the identity of these isolates as Foc, the primary causative agent of wilt in bananas. The morpho-cultural characterization of Foc isolates showed large variations in colony morphological features, intensity, and pattern of pigmentation, chlamydospores, and conidial size. The molecular identification of these isolates using Race1- and Race4-specific primers established their identity as Race1 of Foc, with the absence of Tropical Race 4 of Foc. For a more comprehensive understanding of the genetic diversity of Foc isolates, we employed ISSR molecular typing, which revealed five major clusters. About 96% of the diversity within the Foc population indicated the presence of polymorphic loci in individuals of a given population evident from the results of Nei’s genetic diversity, Shannon’s information index, and the polymorphism information content values, apart from the analysis of molecular variance (AMOVA). The current findings provide significant insights toward the detection of Foc variants and, consequently, the deployment of effective management practices to keep the possible epidemic development of disease under control along the Malbhog banana growing belts of northeast India. Full article

Peach (Prunus persica) is a temperate fruit crop that is widely cultivated in the northern region of Vietnam. This study investigated the genetic diversity of 59 peach accessions collected from northern mountainous provinces in Vietnam using ISSR (inter-simple sequence repeat) markers. The obtained results enabled the evaluation of genetic variation and relationships among peach varieties, which supports breeding programs and germplasm conservation. The analysis identified high levels of polymorphism (92.5%) across six ISSR primers. The accessions were grouped into two main clusters based on a genetic similarity coefficient threshold of 0.674. There were no significant correlations between genetic and geographic distances. The findings underscore the importance of molecular markers like ISSR for identifying genetic relationships and conserving germplasm resources. The results also highlight the potential genetic drift resulting from the trading and exchange of peach varieties among farmers, leading to the creation of regionally named varieties. This study provides valuable insights into the genetic diversity of Vietnamese peaches, supporting efforts to preserve and utilize these resources for breeding and agricultural development. Full article