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Berberis vulgaris L. is a shrub exhibiting significant morphological and biochemical diversity. This study evaluated nine wild genotypes from Isfahan Province, Iran, based on 21 morphological and biochemical traits. Fruit length varied from 0.6 to 1.07 cm, width from 0.47 to 0.72 cm, and weight from 0.14 to 0.26 g, with colors ranging from light yellow to dark red. Pearson correlation analysis revealed key relationships: fruit weight correlated positively with seed length ( r  = 0.50), fruit color had a strong negative correlation with fruit length ( r = − 0.68) and a positive correlation with leaf color ( r  = 0.74). The number of fruits per panicle correlated negatively with seed width ( r = − 0.73), while fresh fruit weight correlated positively with leaf blade length ( r  = 0.79). Biochemical analysis revealed significant antioxidant activity (ranging from 89.20 to 97.10%), which correlated with phenolic content (5.18–14.34 mg GAE/g DW) and flavonoid content (5.3–16.86 mg QE/g DW). Sample 2 had the highest total phenolics (TP) content (14.34 mg GAE/g DW.), flavonoids (13.83 mg QE/g DW), whereas sample 7 exhibited the highest flavonoid concentration (16.86 mg QE/g DW) and sample 3 showed the strong antioxidant capacity (97.10%). Hierarchical clustering grouped genotypes into three clusters: Cluster I (samples 7 and 8) with distinct biochemical profiles; Cluster II (samples 1 and 2) with darker fruits and higher phenolics; Cluster III with lighter fruits and lower biochemical levels. These results highlight the genetic diversity and medicinal potential of B. vulgaris , suggesting fruit color and morphological traits as useful markers for selecting genotypes with high antioxidant capacity. Conservation and breeding efforts should prioritize these valuable wild resources for pharmaceutical and nutraceutical uses. Future studies integrating molecular markers with morphological and biochemical traits will provide deeper insights into the genetic basis of variability and support targeted breeding programs.

Melatonin is a regulatory molecule that increases secondary metabolite biosynthesis in plants. Berberis vulgaris L. is a well-known medicinal plant, valued for its alkaloids and phenolic compounds. This study aimed to determine whether and how melatonin affects biosynthesis of alkaloids (particularly berberine) and phenolics in B. vulgaris . For this, cell cultures were treated with 0, 25, 50, and 100 µM melatonin, and the contents of berberine, phenolics, and amino acids, activity of phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL), and oxidative status were assessed. Melatonin altered total alkaloid and berberine contents, with the highest level observed at 50 µM (1.63 mg g −1 FW and 60.63 µg g −1 DW, respectively). Melatonin also significantly increased tyrosine (Tyr) level, a precursor to berberine. Besides, phenylalanine (Phe) content and PAL activity enhanced in response to exogenous melatonin, leading to a dose-dependent cinnamic acid and coumaric acid accumulation. The HPLC results revealed that melatonin also influences the individual flavonoids profile. The highest increase was detected in catechin, kaempferol, and myricetin contents, which were 19.59, 14.73, and 9.26 times higher than the control levels, respectively. Oxidative balance was also disturbed in melatonin-treated B. vulgaris cells, indicated by increased generation of hydrogen peroxide (H 2 O 2 ) and nitric oxide (NO). The resulting oxidative stress was controlled by activation of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX), along with increased ascorbic acid (AsA) levels. These findings suggest that melatonin can allocate Tyr and Phe towards berberine and phenolics accumulation, respectively, possibly by modulating the oxidative status.

Native to Iran, seedless barberry ( Berberis vulgaris L.) is a nutritionally and economically important crop prized for its therapeutic uses and food industry applications. Nevertheless, the vegetative propagation of the plant and lack of seeds limit conventional breeding techniques, therefore reducing genetic variation and impeding cultivars improvement. This work sought to maximize gamma radiation dosages to cause mutations in seedless barberry hence increasing genetic variability for breeding projects. Hardwood cuttings were subjected to gamma radiation dosages of 0 (control), 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 Gy using a Cobalt-60 source. Under both in vitro and in vivo settings, the impacts on survival rates, leaf and shoot development, and morphometric features were assessed. Results revealed that rising radiation doses significantly reduced survival rates and growth metrics. The LD 50 (Lethal dose) was determined to be roughly 19 Gy for the in vivo and 13.6 Gy for the in vitro. Gamma irradiation negatively influenced plant growth according to values for leaf length, leaf width, fresh weight, and dry weight growth reduction (GR 50 ). Optimal dosages for causing mutations, while preserving survival, were found to be 15 Gy for the in vivo and 10 Gy for the in vitro.

Background One of the wild fruit species with a natural distribution in Türkiye, and historically used for medicinal purposes due to its rich composition, is Berberis crataegina DC. Various parts of the plant, including its roots, bark, leaves, flowers, and fruits, have been utilized in traditional medicine, while its fruits are also consumed in various forms as food. This study aimed to characterize the morphological, biochemical, and molecular traits of B. crataegina genotypes naturally growing in the Kayseri region, located in central Türkiye. Results The fruit weight of the genotypes ranged from 0.047 to 0.137 g, fruit width from 3.06 to 4.64 mm, and fruit length from 5.80 to 9.05 mm. Similarly, the leaf traits of the genotypes exhibited wide variation. The total phenolic content ranged from 190.53 to 297.55 mg GAE/100 g, total flavonoid content from 82.03 to 203.89 mg QE/100 g, total anthocyanin content from 4.54 to 11.76 mg cyn-3 gluc/100 g, and total antioxidant capacity between 57.76 and 87.93%. A principal component analysis (PCA) of 11 traits identified four principal components with eigenvalues greater than 1. The first four components accounted for 71.89% of the total variation, with PCA1 explaining 23.48%, PCA2 18.68%, PCA3 16.39%, and PCA4 13.34%. ISSR molecular analysis using nine markers revealed a band count ranging from 4 to 13, polymorphic band count between 3 and 10, and polymorphism rates from 61.54 to 100%, with band lengths ranging from 200 to 1000 base pairs. According to the UPGMA dendrogram based on molecular analyses, the genetic similarity between the genotypes ranged from 0.64 to 1.00, with B2 and B13 being the most similar genotypes. Conclusions In conclusion, the Kayseri region is rich in B. crataegina genotypes with wide genetic variation. The genotypes identified in this region may serve as valuable genetic resources for future studies.

Berberis has numerous applications in the food industry and traditional medicine because of its antioxidant, anti-inflammatory, antimicrobial, and cholagogic properties. In the current study, delimitation of four closely related species (i.e., B. crataegina , B. integerrima , B . orthobotrys and B . vulgaris ; in total 96 accessions) and the genetic variation of the Iranian B. integerrima (59 accessions) were studied using DNA barcodes and a phytochemical approach. A total of 240 sequences of six nuclear and chloroplast DNA barcodes, including ITS, mat K, ndh F, rbc L, trn H- psb A, and trn L- trn F intergenic spacers, were obtained and analyzed using ML, STRUCTURE, UPGMA, AMOVA, and PCoA approaches. Based on our findings, the evaluated DNA barcodes, either individually or in combination, do not provide sufficient resolution for discriminating the closely related species of Berberis . However, ITS and trn H- psb A did get better resolution compared to the other markers. In order to evaluate the genetic diversity of Iranian B . integerrima , we studied 14 wild populations to build the first local DNA barcode reference library. The results of STRUCTURE analysis suggested that B . integerrima accessions can be grouped into four subpopulations. In correlation with the results of STRUCTURE analysis, the studied accessions were resolved into four major clusters on the UPGMA dendrogram, where the accessions from the same or neighboring areas were mostly grouped in a common cluster. The AMOVA analysis revealed that 24.91% (nrDNA) and 34.91% (cpDNA) of the observed variances corresponded to differences among subpopulations, whereas 75.08% (nrDNA) and 65.09% (cpDNA) matched to differences within subpopulations. For the phytochemical analysis, six key traits (total phenolic content (TPC), total flavonoid content (TFC), anthocyanin content, and antioxidant activity using DPPH, FRAP, and ABTS⁺ assays) were evaluated across 25 populations of B . integerrima . Moreover, we observed significant variation in phytochemical content among B. integerrima populations. The accessions that grow at higher elevations have the most antioxidants (DPPH = 90.62%, FRAP = 4.62 mg GAE g⁻ 1 DW), anthocyanins (5.06 mg g⁻ 1 DW), TPC (36.38 mg GAE g⁻ 1 DW), and TFC (8.52 mg QE g⁻ 1 DW). We identified positive correlations between these phytochemicals and antioxidant activity. Cooler climates, higher latitudes, increased UV radiation, and lower rainfall enhanced bioactive compound levels, indicating environmental stress stimulates secondary metabolite production. These findings underscore the significant nutraceutical and medicinal potential of populations growing at higher altitudes. To our knowledge, this study represents the most comprehensive analysis of the genetic variation and population structure of barberry in Iran. The insights gained from this work offer valuable implications for germplasm collection, genetic improvement, and the systematic utilization of B . integerrima .

Medicinal plant extracts are typically interpreted mainly through their organic phytochemical constituents; however, growing evidence suggests that inorganic components may also contribute to biological activity and cellular fate. In this study, we performed an integrated organic–inorganic mechanistic evaluation of the anticancer activity of Berberis thunbergii DC. leaf extract, focusing on its effects in A549 human lung carcinoma cells. Treatment concentrations were determined using MTT-based cytotoxicity assays. The phenolic composition of the methanolic extract was characterized by LC–MS/MS, while apoptosis induction and cell-cycle distribution were analyzed by flow cytometry. Inorganic anions were quantified by ion chromatography under conditions corresponding to the A549 IC50 value. LC–MS/MS profiling identified rutin, chlorogenic acid, and hesperidin as the major phenolic constituents. Exposure of A549 cells to the extract at its IC50 significantly reduced cell viability, whereas HEL299 cells retained high viability. Ion chromatography revealed the presence of fluoride, phosphate, and sulfate anions. Functional assays using certified NaF, Na 2 HPO 4 , and Na 2 SO 4 standards showed that NaF significantly induced apoptosis and G0/G1 arrest, while sulfate caused S-phase depletion and G2/M accumulation. No significant effects were observed after phosphate treatment. These phase-specific responses suggest that Berberis thunbergii DC. leaf extract acts as a phytochemical–ionic system in which inorganic anions contribute to selective cell-cycle modulation in A549 cells.

Evolutionary classification attempts to integrate information on shared ancestry, evolutionary processes, and phenetic information into the taxonomic concept. Berberis species in Azad Jammu and Kashmir, especially the Poonch division are highly diverse. However, a comprehensive study was required to reveal the evolutionary relationship between the Berberis species. Hence, we planned our study to explore the evolutionary relationship between Berberis species through morphological traits. For this purpose, eight identified species i.e., Berberis lycium , Berberis parkeriana , Berberis ulcinia , Berberis royleana , Berberis brevissima , Berberis kashmiriana , Berberis orthobotrys, and Berberis orthobotrys subsp. capitata were identified and subjected to phylogenetic characterization. Pearson correlation coefficients indicated a strong correlation of fruit yield per plant with the number of primary branches per plant and the number of berries per branch. Principal component analysis biplot showed that B. lycium , B. parkeriana , B. brevissima, and B. orthobotrys were intended for most of the variation within the study area. Cluster analysis identified morphological similarities of B. lycium with B. parkeriana , B. brevissima with B. royleana, and B. orthobotrys with B. orthobotrys subsp. capitata . Our research confirmed the previous predictions that B. lycium is closely associated with B. parkeriana and B. orthobotrys subsp. capitata is the sub-specie of B. orthobotrys . The research contributed to the effective identification and evolutionary classification of Berberis species found in the area. The study provided an effective tool to advance the knowledge of species identification, characterization, and evolution in the region. These findings are important for future identification, conservation of Berberis species for medicinal use, and wheat rust disease management research.

Berberis integerrima Bunge is a valuable plant of the Berberidaceae family used in medicine and industry. The properties of the different parts of this plant vary and depend on the distribution of its secondary metabolites, including alkaloids and phenolic compounds. Here, we aimed to evaluate and compare phenolics and alkaloids of the different organs and tissues of B. integerrima . Our results showed that the highest content of phenolic compounds was found in the fruit and leaf, while the total alkaloid level was higher in the root. In the fruit, leaf and stem, the main phenolic acids were caffeic acid, cinnamic acid and gallic acid, respectively. In contrast, the highest levels of ferulic acid, catechin, resveratrol and luteolin were detected in the root. The highest content of berberine, one of the most important alkaloids of barberry, was found in the root, especially in the bark tissue. Further experiments showed that phenolic compounds and berberine, in the aerial organs and root of B. integerrima , respectively, are likely responsible for the antioxidant capacity of these organs. Given the high berberine content of the root (6.26 mg g -1 Dry Weight), and after trying to find a simple yet effective method to extract berberine, it was found that 80% ethanol containing 2% acetic acid at 25 °C with 72 h of maceration gave the highest berberine yield. Overall, the distribution and accumulation patterns of the secondary metabolites in the different organs of B. integerrima lead to their different applications.

Berberis darwinii, known for its bioactive alkaloids like berberine and palmatine, has gained attention for its medicinal properties. However, comprehensive studies on the specific bioactive molecules of Michay are lacking, as previous research has primarily focused on wild plants. Therefore, this study proposes to evaluate the alkaloid content in various tissues of B. darwinii collected from different locations, aiming to identify high-yielding accessions suitable for consistent bioactive alkaloid production. This research focuses on plants from a cultivated Michay orchard established five years ago. Leaves, stems, roots, and fruits from 96 accessions of Michay were collected to obtain an alkaloidal extract used for the characterization and comprehensive analysis of bioactive alkaloids through high-performance liquid chromatography. Based on these results, a search for the main outliers was conducted to identify the accessions with the highest alkaloid production. The results showed that roots had the highest concentrations of both berberine and palmatine, followed by stems, while leaves and seeds had lower levels, and the pulp from fruits had no detectable alkaloids. Notably, alkaloid concentrations reached up to 30,806 µg/g in roots, with accession C2P18 standing out for its combined total of 20,827.74 µg/g of berberine and 9978.27 µg/g of palmatine. Accession C3P26 showed the highest berberine concentration at 26,482.20 µg/g. These values underscore the wide variation in alkaloid accumulation and highlight the potential for selecting elite accessions with exceptionally high yields. These findings highlight the importance of plant selection for optimal alkaloid extraction. Choosing high-yielding accessions and standardizing cultivation practices will ensure a stable supply of berberine and palmatine for pharmaceutical, nutraceutical, and food industry applications.

In the South Khorasan province of Iran, the seedless barberry variety 'Zereshk Bidaneh' is cultivated extensively and is noted for its exceptional resilience to adverse climatic conditions. As part of a breeding program to develop new barberry cultivars, 90 F hybrid progenies were produced from crosses between 'Zereshk Bidaneh' and selected wild genotypes. This study investigates the biochemical and phenological diversity within this F population. Phenological assessments revealed a broad range in flowering time among genotypes, with statistically significant differences (p < 0.05). The earliest flowering was observed on April 15 in genotype '0506', while the latest occurred on May 5 in '0202', resulting in a 40-day interval. Biochemical evaluations indicated substantial and significant variability (p < 0.05) in total anthocyanin content (TAC), with values spanning from 133.37 mg/L ('0203') to 2063.55 mg/L ('0601'). The average TAC was 614.44 mg/L, and the high coefficient of variation (67%) emphasized the extent of diversity. Notably, four genotypes ('0601', '0606', '0607', and '0617') exhibited TAC levels above 1250 mg/L. Vitamin C concentrations also showed significant differences (p < 0.05), averaging 108.50 mg/100 mL, with several genotypes exceeding 130 mg/100 mL. Total antioxidant activity (TAA) ranged markedly from 7.89 to 90%, while total phenolic content (TPC) had a mean of 1770 mg GAE/100 mL (p < 0.05), underscoring the presence of bioactive compound-rich genotypes. Total soluble solids (TSS) varied between 14 and 38 °Brix, with a mean of 29.27 °Brix (p < 0.05). Measured pH values ranged from 2.66 to 3.76. The fruit flavor index (FI) also varied widely, ranging from 0.98 to 7.68 (p < 0.05). Principal component analysis (PCA) revealed that the first two principal components accounted for 50.04% of the total variance, with TAC, TAA, titratable acidity (TA), and FI identified as the most influential variables. Heat map analysis further classified the genotypes into seven distinct biochemical clusters, reinforcing the diversity patterns observed. The results reveal extensive phenotypic and biochemical diversity among the F barberry progenies, with several traits showing statistically significant variation (p < 0.05). These findings provide a valuable foundation for selecting superior genotypes and advancing breeding programs aimed at developing improved barberry cultivars with enhanced nutritional and horticultural qualities. Among the evaluated progenies, genotypes such as '0601' and '0203' stood out due to their exceptionally high total anthocyanin content (2063.55 mg/L) and antioxidant activity (90%), respectively, highlighting their potential value in future breeding strategies.