Search Results (188)

This study presents an integrated approach combining molecular, phytochemical, and biological analyses to characterize a newly discovered Zizania specimen from the northern Nile Delta, Egypt. Genetic fingerprinting using RAPD and ISSR markers revealed 85% band-sharing similarity with Zizania texana (Z. texana), though distinct morphological and genetic traits suggested potential intraspecific variation. Phytochemical profiling identified high concentrations of bioactive compounds, including quercetin (42.1 µg/mL), β-caryophyllene (11.21%), and gallic acid (23.4 µg/mL), which are pertinent and correlated with robust biological activities. The ethanolic leaf extract exhibited significant antioxidant capacity (IC₅₀ = 38.6 µg/mL in DPPH assay), potent antimicrobial effects against Candida albicans (C. albicans) (IC₅₀ = 4.9 ± 0.6 µg/mL), and dose-dependent cytotoxicity against cancer cell lines. MCF-7 has the lowest IC₅₀ (28.3 ± 1.5 µg/mL), indicating the highest potency among the tested cell lines. In contrast, HepG2 demonstrates moderate sensitivity (IC₅₀ = 31.4 ± 1.8 µg/mL), while A549 shows the highest IC₅₀ value (36.9 ± 2.0 µg/mL), indicating greater resistance. These findings underscore the taxonomic novelty of the specimen and its potential as a source of natural antioxidants, antimicrobials, and anticancer agents. The study highlights the importance of interdisciplinary approaches in resolving taxonomic uncertainties and unlocking the medicinal value of understudied aquatic plants. Full article

Longshan Lilium lancifolium is a well-known medicinal and edible lily and has been registered as a geographical indicator in China. Polyploidization confers many advantages in lily production; however, characteristics of Longshan L. lancifolium improved by polyploidization have not been reported. Here, polyploidization was induced in regenerated Longshan L. lancifolium shoots using colchicine, and the mutant plantlets were characterized by morphological observation, flow cytometry, and inter simple sequence repeat (ISSR) marker technology. The optimal medium for inducing shoot regeneration was Murashige and Skoog (MS) media supplemented with 0.2 mg/L of naphthaleneacetic acid (NAA) and 0.4 mg/L of thidiazuron (TDZ). The greatest mutation induction effect was obtained after soaking the regenerated shoots in 0.10% colchicine for 48 h, for an 80.00% frequency of morphological variants. Forty-one mutant plantlets were subjected to flow cytometry, identifying one homozygous polyploid, ‘JD-12’, and one chimeric polyploid, ‘JD-37’. Additionally, 68 chromosomes were found in the ‘JD-12’ root tip cells. Compared with the control, both the tissue-cultured and field-generated ‘JD-12’ plantlets presented a slight decrease in plant height, a darker green leaf color, a rougher leaf surface, and a larger bulblet diameter; furthermore, the upper epidermal and guard cells of ‘JD-12’ were much larger with a significantly lower stomatal density. The ISSR marker detection indicated a genetic variation rate of 6.10% in ‘JD-12’. These results provide a basis for lily polyploidization breeding and the cultivation of superior Longshan L. lancifolium via shoot regeneration. 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

Background/Objectives: Saussurea obvallata (DC.) Edgew., Asteraceae, is a traditional medicinal herbnative to the Qinghai–Tibet Plateau (QTP). Pharmacological investigationshave validated its pharmacological effects in anti-tumor, anti-inflammatory, heat-clearing, detoxifying, and analgesia. S. obv is presently facing habitat fragmentation and population decline. Therefore, we analyzed its genetic and chemical diversity to provide a scientific basis for the conservation and sustainable use of S. obv. Methods: Seven populations of S. obv were sampled from Xizang, China. The genetic diversity was analyzed using inter-simple sequence repeat (ISSR) markers, and metabolites were identified by ultra-high-performance liquid chromatography-tandem-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). Correlation analysis among genetic diversity, differential metabolites, and climatic factors were performed by R. Results: The genetic diversity among and within populations were both lowly and significantly correlated with geographical distance, showing a decreasing trend from east to west of the QTP. A total of 110 compounds were identified, including flavonoids, phenylpropanoids, lipids, fatty acids, terpenoids, alkaloids, etc. The metabolite contents among populations varied greatly and were related to environmental factors, mainly annual mean temperature and temperature fluctuation. The genetic diversity had little effect on the metabolic differences. Conclusions: These findings provided valuable baseline information for the conservation and pharmacological utilization of S. obv. Meanwhile, further research is necessary for the efficacy evaluation of anti-inflammatory, anti-tumor, radiation protection, and scar removal both in vitro and in vivo. 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

In this study, a reliable and efficient micropropagation protocol was developed for Stachys byzantina, a valuable and promising ornamental species. For the initial in vitro cultures on the Murashige and Skoog (MS) medium, shoot tips were used as explants. The addition of 5 μM of kinetin (KIN) resulted in the production of multiple (6.0 shoots/explant) and elongated (3.6 cm) shoots. The MS medium supplemented with 10 μM of a-Naphthaleneacetic acid (NAA) proved efficient for the in vitro rooting (73.3%) of the microshoots. For the ex vitro rooting of the microshoots, the treatment with 0.5 g L⁻¹ of Indole-3-butyric acid potassium salt (K-IBA), before planting in 1:1 (v/v) peat and perlite substrate and placed in a fog system, led to 86.7% rooting. The acclimatization stage was successful, and 96.7% survival was recorded for the ex vitro-rooted plantlets. Inter Simple Sequence Repeat (ISSR) markers were employed to examine the genetic uniformity of the in vitro-derived plantlets with the mother S. byzantina plants. The monomorphic banding pattern in the micropropagated plants and the mother plant confirmed the genetic uniformity of the in vitro-derived plantlets and revealed the reliability of the proposed in vitro protocol for S. byzantina. As far as we know, this is the first study on a combined micropropagation and genetic uniformity assessment of the species, the findings of which could be further used to apply new in vitro cultivation techniques or to produce elite genotypes of S. byzantina. Full article

Efficient extraction of high-quality DNA from plants is a critical challenge in molecular research, especially in species such as Gossypium barbadense L., native to Peru, due to the presence of inhibitors such as polysaccharides and phenolic compounds. This study presents a modified CTAB-based protocol with silica columns that is designed to overcome these limitations without the need for liquid nitrogen or expensive reagents. Native cotton samples were collected in Lambayeque, Peru, and processed using a simplified procedure that optimizes the purity and concentration of the extracted DNA. Eight cultivars of G. barbadense L. with colored fibers (cream, fifo, light brown, dark brown, orange-brown, reddish, fine reddish, and white) were evaluated, yielding DNA with A260/A280 ratios between 2.14 and 2.19 and A260/A230 ratios between 1.8 and 3.14; these values are higher than those obtained with the classical CTAB method. DNA quality was validated by PCR amplification using ISSR and RAPD molecular markers, which yielded clear and well-defined banding patterns. Furthermore, the extracted DNA was suitable for advanced applications, such as Sanger sequencing, by which high-quality electropherograms were obtained. The results demonstrate that the proposed protocol is an efficient, economical, and adaptable alternative for laboratories with limited resources, allowing the extraction of high-quality DNA from Gossypium barbadense L. and other plant species. This simplified approach facilitates the development of genetic and biotechnological research, contributing to the knowledge and valorization of the genetic resources of Peruvian native cotton. Full article

The taxonomic identification of plant species is traditionally based on morphological traits, the use of which may create difficulties in cases of close-related species showing great morphological variability. In such cases, the use of DNA markers for species identification and delimitation can be of great help. Himantoglossum W.D.J.Koch (Orchidaceae) is a genus with notable morphological variability, comprising the clade hircinum-caprinum (Himantoglossum s.s.) with nine taxa, from which H. jankae, H. hircinum, H. montis-tauri, H. caprinum and H. samariense have being reported in Greece. However, a previous morphological study of Himantoglossum s.s. from all over Greece could not verify the presence of these reported species, but of only one highly diverse taxon throughout the country. Here, we studied the genetic variation and differentiation of Himantoglossum s.s. populations from the entire distribution of the genus in Greece employing ISSR markers, to further elucidate the taxonomic status of Himantoglossum s.s. in Greece. High genetic variation was revealed, both in the populations of the “core” distribution and in the peripheral/marginal ones, pointing to their evolutionary potential. This variation is mainly attributed to differences within the populations and, to a lesser extent, among them. No differentiation of the populations proposed to belong to a different taxon was found and no species-specific markers were identified that may discriminate the above populations from the rest. In addition, two cpDNA and one nDNA fragments (accD, psbA-trnH and ITS2, respectively) were sequenced in a number of individuals representative of the whole dataset. All three fragments were conserved, showing restricted polymorphism and having no correlation to the populations or to the taxa of Himantoglossum s.s. in Greece. Overall, the high genetic variation of the populations of Himantoglossum s.s. in Greece, especially of the peripheral/marginal ones, is a valuable asset towards their conservation. Full article

Moricandia moricandiodes is an endemic species found in the south and east of the Iberian Peninsula. Five subspecies have been recognized, and all exist as fragmented populations on limestones and marls with salt and gypsum intrusions under a continental Mediterranean climate, except for one of the subspecies, which inhabits semi-arid and hotter environments. In this study, we sampled populations covering the distribution area of the species and performed a population and phylogeographic study to assess the evolutionary history of populations and the taxonomic relationships of subspecies. ISSR markers, nrITS, and plastid sequences were used in the analyses. The results revealed that, in general, southern populations showed higher genetic diversity than northern populations, suggesting that the former are located in glacial refugia. Furthermore, we did not find clear differences between subspecies, except for M. moricandioides subsp. pseudofoetida, which showed exclusive haplotypes and an exclusive ribotype. Isolation and rapid divergence are discussed as the probable causes of differentiation, whereas bottlenecks and secondary contact between populations would explain the absence of differentiation among the other subspecies. Finally, we propose a few guidelines for the conservation of M. moricandioides. 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

Tamarix chinensis Lour. is a halophytic shrub native to coastal China, commonly used in afforestation and ecological restoration due to its high tolerance to salinity and drought. To understand how this species maintains genetic variation and adapts to extreme environments, we examined the genetic diversity, mating system, and spatial genetic structure of a natural T. chinensis population on the geographically isolated and environmentally harsh Wudi Seashell Island. Using both SSR and ISSR markers, we observed high levels of genetic diversity despite the small population size and spatial fragmentation. SSR markers revealed an average of 11.75 alleles per locus, with an expected heterozygosity (He) of 0.754 and an observed heterozygosity (Ho) of 0.702. ISSR markers showed a polymorphic locus rate of 97.87%, with a mean He of 0.402. Parentage analysis revealed relatively long seed and pollen dispersal distances, with most dispersal occurring within 150 m and seeds and pollens occasionally reaching 948 m and 447 m, respectively. The species exhibited a mixed mating system, with a multilocus outcrossing rate of 0.554, contributing to gene flow and reducing inbreeding. A fine-scale spatial genetic structure was detected within 75 m, consistent across both SSR and ISSR markers, suggesting limited local gene dispersal. These findings provide new insights into the adaptive strategies of T. chinensis in marginal habitats and offer valuable guidance for conservation and restoration efforts in vulnerable coastal ecosystems. Full article

Honey is a multifaceted substance whose composition is intricately affected by various biotic and abiotic elements generated in the bee colony’s surroundings, including botanical and geographical origins, climatic conditions, soil characteristics, and beekeeping techniques. Monofloral honeys are identified by pollen analysis and are derived from the nectar of a predominant plant species, exhibiting rich sensory and nutritional profiles, making them food matrices with unique characteristics and excellent qualities. To explore the monofloral honey potential harvested in different regions of Portugal, a comprehensive study was conducted including the determination of phenolic composition and the assessment of biological activities. In addition to this evaluation, the inter simple sequence repeat (ISSR) was used to help differentiate honeys by botanical origin. The phenolic content and the antioxidant capacity were evaluated by spectrophotometric methods, observing, in general, differences between monofloral honeys. The honey from Citrus sinensis (Silves) exhibited the lowest phenolic content, including total phenols, ortho-diphenols, and flavonoids, whereas honeydew (Vinhais) showed the highest values. Regarding the antioxidant capacity, honey from Lavandula stoechas (Almodôvar) presented the lowest values, while honeydew (Vinhais) displayed the highest values for both DPPH and FRAP assays. In relation to the ABTS assay, the honey from Metrosideros excelsa (Aveiro) exhibited the lowest values, whereas the honey from Eucalyptus spp. (Arouca) showed the highest. The ISSR marker analysis allows the distribution of the samples based on the honey’s botanical origin, suggesting its potential role in honey authentication. Full article

Garlic (Allium sativum L.) is the second most significant species within the Allium genus worldwide, widely used in cooking and both traditional and modern medicine due to its beneficial biological and therapeutic properties. In Italy, diverse pedo-climatic conditions and historical–cultural fragmentation have led to the development of various garlic landraces, prized for their unique organoleptic qualities and cultural importance. This study aimed to assess the intra-varietal diversity and uniqueness of two red garlic landraces from the Lazio region in central Italy, “Aglio Rosso di Castelliri” and “Aglio Rosso di Proceno”, using SSR and ISSR molecular markers, along with evaluations of bulb morphological traits, total phenolic content, and antioxidant properties. The molecular analysis included 11 accessions of “Aglio Rosso di Castelliri”, nine of “Aglio Rosso di Proceno”, and 15 control accessions, comprising eight Italian red-type garlic landraces, four Spanish red garlic commercial varieties, two white garlic accessions, and an accession of A. ampeloprasum var. holmense used as an outgroup. SSR and ISSR markers revealed moderate genetic diversity within the collection, with mean PIC values of 0.41 and 0.17, respectively. The molecular data identified four distinct genetic clusters, with the two Lazio landraces forming separate groups, indicating their genetic distinctiveness. The results from the STRUCTURE analysis support the hypothesis that these landraces may have originated from the widely cultivated “Aglio Rosso di Sulmona” or a common ancestral population once prevalent in central Italy. The study also revealed significant intra-population genetic diversity within the two garlic landraces, underscoring the need for in situ conservation and clonal selection. Phenotypic evaluations confirmed the distinctiveness of the two landraces, with “Aglio Rosso di Castelliri” characterized by smaller bulbs and cloves with higher dry matter content and distinct color profiles. Additionally, significant variation in total phenolic content and antioxidant activity was observed by analyzing 13 selected accessions from the two landraces (six from “Aglio Rosso di Proceno” and seven from “Aglio Rosso di Castelliri”) and five red garlic control accessions, with the two Lazio landraces exhibiting higher levels than the control group. This study highlights the importance of integrating molecular, phenotypic, and chemical analyses to understand garlic landrace diversity, with significant implications for their conservation and protection of local agro-food products. Full article

Apomictic populations, which produce seeds with embryos without proper sexual syngamy, often show low genetic diversity, but eventually, such diversity has been reported to be surprisingly high. We studied here the genetic diversity in agamic complexes of Eriotheca crenulata (comb. n. E. gracilipes), E. pubescens (Malvaceae-Bombacoideae), and Handroanthus ochraceus (Bignoniaceae), tropical tree species from the savannas in Central Brazil. We evaluated the genetic diversity and structure of self-fertile polyploid sporophytic apomicts versus self-sterile diploid or tetraploid sexual populations by using dominant ISSR markers. Genetic diversity was either similar or even higher in apomictic populations of E. crenulata and E. pubescens, but the opposite was observed in some populations of H. ochraceus. Only two individuals of E. pubescens showed identical ISSR profiles, so strict clonality in adult individuals was very rare among the studied trees. The genetic variability was notably higher within populations than among populations of H. ochraceus and very similar among and within populations of Eriotheca species. Ordination, clustering, and Bayesian analyses showed a clear distinction between populations of Eriotheca species with different breeding systems. But for H. ochraceus, a sexual population was actually grouped with the apomictics. As in other studies, eventual sexual and recombination events seem to increase genetic diversity in apomictic populations. This may explain the similar genetic diversity among apomictic and sexual populations in the studied agamic complexes and the virtual absence of strict clonal individuals. The results have evolutionary and ecological consequences for the threatened Neotropical savanna trees. Full article