Search Results (5,329)

Black rice is rich in anthocyanins with potential antioxidant benefits, but their specific role in storage stability remains unclear due to confounding genetic backgrounds in previous studies. In this study, we used CRISPR/Cas9-mediated gene editing to generate OsKala4 knockout lines in the black rice cultivar Heizhen (HZ), creating an isogenic system to test whether endogenous anthocyanins contribute to storage-related quality stability. Knockout lines showed blocked anthocyanin biosynthesis (0.5–0.6 vs. 155.6 mg/100 g, p < 0.001) and altered grain composition. Under accelerated aging (45 °C, 90% RH, 2 weeks), HZ maintained higher antioxidant capacity (p < 0.05) and exhibited less pronounced starch aging than the representative knockout line KO2. Apparent amylose content increased less in HZ than in KO2 (16.7% vs. 28.1%, p < 0.05). HZ also showed smaller changes in pasting, thermal, and structural properties. XRD and FTIR analyses further suggested better maintenance of starch crystallinity and molecular order in HZ under accelerated aging conditions. These results suggest that endogenous anthocyanins were associated with storage-related quality stability in black rice. However, direct mechanistic evidence and validation under natural storage conditions are still needed. Full article

The valorization of agri-food by-products aligns with circular economy principles and offers sustainable sources of bioactive compounds. This study investigated the peels of the purple-fleshed Solanum tuberosum L. cv. Vitelotte Noire (VN), cultivated in Sardinia, as a potential resource for nutraceutical antioxidants. Extracts were obtained using solvents of different polarities (water, 80% and 96% ethanol) and characterized. Phytochemical screening revealed high concentrations of total phenolics, flavonoids, and anthocyanins, with the 96% ethanolic extract showing superior anthocyanin content. Antioxidant capacity, assessed via ORAC-PYR, TEAC-ABTS, and DPPH assays, was highest in the alcoholic extracts. Furthermore, all extracts showed protective effects in an in vitro model of AAPH-induced oxidative DNA damage, as indicated by the preservation of plasmid supercoiling. Untargeted LC-QTOF-MS analysis detailed a rich metabolomic profile, including organic acids, amino acids, and vitamins. The findings confirm VN peel as a potent, sustainable source of antioxidants, supporting its valorization for developing high-added-value nutraceutical and functional food ingredients, while reducing waste disposal costs and environmental impact. Full article

Background/Objectives: Anthocyanins are dietary pigments associated with reduced risk of chronic diseases, yet their low systemic bioavailability challenges the traditional direct antioxidant hypothesis. This review aims to reconceptualize anthocyanin bioactivity by proposing the gut microbiome as a key mediator that biotransforms these compounds into bioactive metabolites responsible for systemic health effects. Methods: This review synthesizes evidence on the microbial metabolism of anthocyanins and includes a structured appraisal of the literature using an evidence evaluation framework analogous to GRADE, focusing on their transit to the colon, enzymatic biotransformation by gut microbiota, and resulting production of phenolic metabolites such as protocatechuic acid (PCA). It also examines the role of specific bacterial taxa (e.g., Bifidobacterium and Lactobacillus) in enhancing bioavailability and explores the downstream cellular pathways modulated by these metabolites. Results: Gut microbiota convert anthocyanins into smaller phenolic metabolites such as PCA, syringic acid, gallic acid, and other respective metabolites, which achieve plasma concentrations up to 100-fold higher than parent compounds and can cross the blood–brain barrier. These metabolites exert systemic effects by modulating key signaling pathways (NF-κB and Nrf2) and restoring redox homeostasis. Additionally, beneficial gut bacteria enhance anthocyanin bioavailability and support the production of short-chain fatty acids (SCFAs). Conclusions: Systemic health benefits of anthocyanins are largely mediated by gut microbiota through the generation of bioactive metabolites. This microbiota-driven process redefines the mechanistic understanding of anthocyanin action and highlights the microbiome as a critical determinant of their efficacy in preventing cardiometabolic and neurodegenerative diseases. Full article

Despite the wide and accepted implementation of contemporary pharmaceutical medicine, the use of medicinal plants still prevails in several regions around the world, including Mexico. According to the World Health Organization (WHO), the use of incorrect species in natural and complementary medicine is a threat to consumer safety. Therefore, there is a need to characterize properly those plant species used in traditional medicine. In this study, a medicinal plant called Calanca, which is traded in the local market of a small community within the State of Puebla (Mexico), was characterized by different approaches. Conventional and molecular taxonomy analyses showed that Calanca belonged to the Asteraceae family, genus Chrysactinia. On one hand, molecular markers (rbcL, matK and ITS) helped to identify Calanca at the species level, being identified as C. mexicana. On the other hand, although not used for molecular taxonomy, additional gene markers were amplified and submitted to the GenBank database to expand the toolkit for C. mexicana identification. In addition, soil taxonomy and quantitative chemical analyses provided insights into the relationship between growing conditions and the chemical compounds produced by C. mexicana. Chemical compounds associated with medicinal properties such as phenolic acids, flavonoids, terpenes, and anthocyanins were identified in C. mexicana extracts. Finally, greenhouse conditions for the cultivation of this species were also investigated. Overall, this comprehensive characterization provides the essential botanical and chemical foundation required for future toxicological and clinical safety assessments, while establishing a robust framework for the long-term conservation of this endangered medicinal resource. Full article

This study provides a comprehensive evaluation of the phytochemical composition, antioxidant capacity, antibacterial activity, and mineral content of Trigonella foenum-graecum, Linum usitatissimum, and Panicum miliaceum extracts obtained using aqueous, ethanolic, and methanolic solvents. An integrated analytical strategy combining LC–MS/MS-based metabolite profiling, mineral analysis, and multivariate statistical tools (PCA) was applied to investigate the relationships between chemical composition and biological activities. The ethanolic extract of P. miliaceum showed the highest total phenolic content (TPC: 157.438 ± 0.521 µg GAE/mg extract), whereas L. usitatissimum exhibited the strongest antioxidant activity (IC₅₀ ≈ 65 µg/mL). Trigonella foenum-graecum displayed the most significant antibacterial activity, with a minimum inhibitory concentration (MIC) of 62.5 mg/mL against Staphylococcus aureus. LC–MS/MS analysis allowed the identification and structural characterization of more than twenty bioactive compounds through multiple reaction monitoring (MRM), including flavonoids, phenolic acids, and anthocyanins. Principal Component Analysis indicated that sample discrimination was mainly driven by solvent polarity rather than plant species, underlining the critical influence of extraction conditions on phytochemical profiles and associated bioactivities. These findings highlight the relevance of combining analytical and statistical approaches to better understand the interplay between plant origin, extraction conditions, and biological properties, and support the potential of these species as promising sources of nutraceutical and pharmaceutical compounds. Full article

Brassica juncea exhibits diverse foliar pigmentation patterns caused by anthocyanin accumulation, but the molecular basis of margin-specific pigmentation remains unclear. Here, we combined anthocyanin measurement, comparative transcriptomics, and functional analysis of BjMYB113 to investigate anthocyanin spatial distribution in mustard. The data showed that anthocyanin content was significantly higher in the leaf margin (LM) than in the leaf interior (LI) of the bicolored accession ZD30. Transcriptome analysis identified 618 DEGs between LM and LI in ZD30, compared with only 134 DEGs in the uniformly purple accession JCS53. Enrichment analyses indicated that ZD30-specific DEGs were mainly involved in flavonoid metabolism, anthocyanin biosynthesis, and secondary metabolism. Expression profiles of genes involved in anthocyanin biosynthesis indicated that BjMYB113 and BjTT8 were more highly expressed in the pigmented margin of ZD30, together with key late biosynthetic genes (DFR, ANS, and UFGT) and GSTF. In addition, transient overexpression of BjMYB113 promoted anthocyanin accumulation in leaves, suggesting that BjMYB113 acts as a positive regulator of anthocyanin accumulation and supporting a putative model in which localized activation of anthocyanin-related genes contributes to margin-specific pigmentation in B. juncea. This study provides insight into the transcriptional regulation of anthocyanin spatial distribution in mustard. Full article

Dihydroflavonol 4-reductase (DFR) plays a pivotal role in regulating flavonoid and anthocyanin biosynthesis, governing the accumulation of plant secondary metabolites. This study aimed to characterize the DFR gene family in Ginkgo biloba and elucidate the function of the predominant gene GbDFR2 in the flavonoid metabolic network. Through transcriptome analysis, three differentially expressed GbDFR genes were identified. Bioinformatic analysis revealed that all three GbDFR proteins are hydrophilic and acidic and belong to the NADB_Rossmann superfamily. RT-qPCR analysis of different tissues of ginkgo revealed that all three GbDFR genes exhibited the highest expression levels in the leaves. An overexpression vector of GbDFR2 was constructed and stably transformed into Nicotiana benthamiana. Metabolomic and qPCR analyses showed that heterologous GbDFR2 expression significantly remodeled the flavonoid profile, upregulating sakuranetin and 3,7-Di-O-methylquercetin while downregulating narcissin and naringenin chalcone. Additionally, it upregulated endogenous NbCHI and NbDFR, and suppressed the transcription factors NbMYL2b and NbERF4a. These findings suggest that GbDFR2 can act as a regulator of flavonol biosynthesis and provide a candidate gene for the metabolic engineering of flavonoids in woody plants. Full article

Wine phenolic composition is strongly influenced by tannin structure, yet how the polymerization degree of exogenous proanthocyanidins modulates wine quality during aging remains unclear. This study investigated the effects of adding grape seed proanthocyanidins (GSP) with different mean degrees of polymerization (mDP 4.63, 3.29, and 1.31) to Cabernet Sauvignon wine by analyzing phenolic compounds, tannin structure, anthocyanin components, CIELAB color parameters, and astringency over 6 months of bottle aging. Low-mDP GSP (rich in galloylated monomers) provided the biggest initial phenolic boost, while high-mDP GSP (dominated by non-galloylated units) sustained tannin enrichment throughout aging. Low-mDP GSP accelerated tannin maturation and color evolution toward aged wine characteristics, with Mv-3-Coglu identified as a key precursor for brick-red hue development. Sensory evaluation revealed that high-mDP GSP enhanced coarse and drying astringency, whereas low-mDP GSP promoted velvety mouthfeel. These findings establish that GSP polymerization degree critically determines phenolic evolution, color stability, and mouthfeel during bottle aging, providing a scientific basis for selecting structure-specific proanthocyanidins to achieve targeted wine quality outcomes. Full article

Rhododendron lapponicum (L.) Wahlenb., prized for its vibrant and diverse floral displays, holds significant ornamental and ecological value. However, advances in its molecular breeding have been constrained by the absence of reliable tools for accurate gene expression analysis. A fundamental requirement for such studies is the identification of stable reference genes for qRT-PCR. To date, no systematically validated reference genes exist for normalizing gene expression across R. lapponicum cultivars with diverse flower colors, representing a major technical obstacle to elucidating the molecular mechanisms of color formation. This study aimed to fill this gap by systematically identifying and validating optimal reference genes for petal tissues in six distinct R. lapponicum cultivars. We assessed the expression stability of 11 candidate genes using four independent algorithms and integrated the results via RefFinder. Our comprehensive analysis across multiple algorithms consistently identified RlaEF1-α and RlaACT as the most stably expressed reference genes. Their reliability was robustly validated by normalizing the expression of RlaMYB113, a key anthocyanin regulator; the normalized expression levels showed an extremely significant difference between rose-red and white cultivars (p < 0.001) and produced a coherent, phenotype-correlated profile, in contrast to the distorted patterns obtained with unstable references. This study establishes RlaEF1-α and RlaACT as a precise dual-gene internal control for qRT-PCR. By providing a validated normalization framework, our work enables accurate quantification of color-related genes and directly supports molecular breeding efforts aimed at the targeted development and selection of novel R. lapponicum cultivars with desirable and stable flower colors. Full article

Saline–alkali stress severely limits crop production worldwide. Soybean [Glycine max (L.) Merr.] is particularly sensitive during seed germination, a stage critical for stand establishment. This complex stress environment encompasses two distinct yet equally critical dimensions: neutral salt stress and alkaline salt stress, each eliciting specialized physiological and metabolic responses. Here, a comparative assessment of four genotypes (tolerant: CN16, CN17; sensitive: Williams 82, K18) under 100 mmol/L Na⁺ revealed that alkaline salt stress exerts a significantly more potent inhibitory effect than neutral salt stress. Tolerant cultivars maintained 75–80% germination under alkaline conditions, whereas sensitive ones dropped below 15%, a divergence primarily driven by superior oxidative mitigation capacity. Integrated multi-omics analysis of the tolerant variety CN16 identified stage-specific regulatory shifts: early alkaline salt stress (60 h) triggers extensive transcriptional reprogramming focused on physical barrier reinforcement, including cell walls and lipid remodeling. By 96 h, regulatory modes between the two stress types diverged: neutral salt elicited a transcriptional surge, while alkaline salt transitioned to a metabolically dominant regulation, shifting flux from growth-related isoflavonoids to defense-related anthocyanins. Crucially, this study uncovers the distinct bioenergetic trade-offs governing these responses: whereas adaptation to neutral salt relies on low-energy galactose metabolism, tolerance to alkaline salt demands energy-intensive processes, specifically the active vacuolar compartmentalization of organic acids and anthocyanins for intracellular buffering. This obligatory energy expenditure restricts biomass accumulation, explaining the severe growth penalties observed in complex saline-alkali environments. Finally, the identification of a core regulatory module driven by key genes, including GmPHOT2b, GmLOG, and GmSHMT08, enriches the metabolic regulatory network under saline-alkali stress, providing core targets and a theoretical framework for precisely balancing metabolic expenditure with biomass accumulation in breeding practice. Full article

Low temperatures severely restrict tea plant (Camellia sinensis) growth and yield stability, yet how brassinosteroid (BR) signaling modulates cold acclimation at a systems level remains insufficiently defined. Here, we integrated transcriptomic and UHPLC–MS metabolomic profiling of tea leaves under Control, 24-epibrassinolide (EBR), Cold, and Cold + EBR treatments to delineate BR-potentiated cold responses. Principal component analyses revealed clear treatment-specific separation and tight clustering of biological replicates at both omics levels. Quantitatively, cold stress induced extensive reprogramming (4075 differentially expressed genes (DEGs) and 298 differentially accumulated metabolites (DAMs)), whereas EBR alone exerted relatively modest effects (231 DEGs and 50 DAMs). Notably, EBR under cold conditions further reshaped cold-responsive networks (371 BR-modulated DEGs and 17 BR-modulated DAMs), consistent with a potentiating role for BR signaling. Functional enrichment analyses highlighted phenylpropanoid metabolism and hormone signal transduction as core responsive modules, with coordinated activation of key gateway genes (PAL, C4H, and 4CL) and concurrent engagement of lignin-, flavonoid-, and catechin-associated branches under Cold + EBR. Metabolomic analyses identified flavonoids as the dominant responsive metabolite class (49.31%), particularly anthocyanins and flavonol glycosides. Integrative TF–metabolite–gene correlation networks prioritized WRKY transcription factors (TEA001162, TEA027058) and a UDP-glycosyltransferase gene (TEA025792) as candidate hub genes linking hormone signaling to phenylpropanoid outputs. Collectively, this work provides a systems-level framework of co-regulated transcript–metabolite modules and candidate molecular targets, offering a foundation for functional validation and practical improvement of cold resilience in tea production. Full article

Fruit anthocyanins are primary determinants of color, sensory quality, and nutritional value in grapes; however, their endogenous biosynthesis is governed by complex interactions among genetic, environmental, agronomic, and postharvest factors. This review elaborates recent advances in physiology and molecular biology to clarify the biosynthetic mechanisms in grapes, including the coordinated action of structural enzymes, MYB–bHLH–WD40 regulatory complexes, hormone-mediated signaling pathways, and vacuolar transport processes. Key environmental factors, such as temperature fluctuations, light exposure, water availability, and soil properties, regulate these networks, contributing to significant variation in pigmentation profiles across cultivars and growing regions. Strategic agronomic practices, including canopy management, regulated deficit irrigation, balanced nutrient management, and temperature-mitigation techniques, further influence pigmentation by modifying the microclimate of the fruit zone during development. Based on these mechanistic insights, this review evaluates targeted strategies for enhancing anthocyanin accumulation, highlighting recent progress in genetic improvement through CRISPR/Cas genome editing, transgenic approaches, and marker-assisted selection (MAS), which enable precise modulation of biosynthetic and regulatory genes. Complementary postharvest interventions, such as optimized cold storage, modified-atmosphere packaging, hormonal elicitors, and controlled oxidative technologies, provide additional opportunities to maintain or enhance pigment stability after harvest. Collectively, these advances establish a comprehensive framework linking molecular regulation with practical vineyard, breeding, and postharvest strategies, offering an integrated pathway to improve anthocyanin consistency, berry quality, and the phenolic characteristics of grape-derived products. Full article

The valorization of agro-industrial by-products is of increasing importance within circular economy strategies. Aronia melanocarpa pomace, a by-product of juice production, represents a valuable source of polyphenols with potential applications in food, nutraceutical, and cosmetic formulations. This study aimed to evaluate the effect of different preservation methods on the polyphenolic composition of Aronia pomace and to optimize accelerated solvent extraction (ASE). Pomace samples were subjected to drying, freeze-drying, freezing (−18 °C), and deep freezing (−80 °C). UAE was applied as a rapid screening method for polyphenol extraction, while ASE was used as an advanced technique for efficient recovery of target compounds. ASE parameters, including temperature (40–120 °C), methanol concentration (40–100%), and number of extraction cycles (1–3), were optimized using response surface methodology (RSM) based on a Box–Behnken design. Qualitative and quantitative analyses were performed using UHPLC–MS and HPLC–DAD. The developed models were statistically significant (p < 0.01) with high coefficients of determination (R² = 0.88–0.97). Temperature had a positive effect on phenolic acid extraction but negatively affected anthocyanins due to thermal degradation. Optimal extraction conditions differed between compound groups: phenolic acids were maximized at 120 °C and 75% methanol (two cycles), while anthocyanins required milder conditions (82 °C, 92% methanol, three cycles). Moreover, our study showed that drying significantly reduced the content of several compounds, particularly anthocyanins, whereas low-temperature methods had minimal impact. The results highlight the importance of tailored extraction strategies and support the sustainable utilization of Aronia pomace as a source of bioactive compounds. Full article

Anthocyanins are plant polyphenols widely regarded as antioxidants, yet they can exert concentration-dependent effects and act as pro-oxidants in specific contexts. Although their protective role in stressed testicular cells is established, their impact on Leydig cell steroidogenesis under non-pathological conditions remains poorly understood. Here, we investigated how an anthocyanin-enriched fraction from Callistemon citrinus (0–1.00 μg/mL) affects androgen synthesis in murine TM3 Leydig cells. Cell viability, intracellular ROS, antioxidant capacity, mitochondrial function, androstenedione production, steroidogenic gene expression, and the exometabolome by ¹H-NMR were assessed. The fraction exhibited biphasic, dose-dependent effects. At 0.01 μg/mL, it induced a mitohormetic response, upregulating mitochondrial complexes III and V. Conversely, higher concentrations (0.10–1.00 μg/mL) reduced metabolic activity, increased intracellular ROS, and significantly suppressed androstenedione synthesis independently of Star. These concentrations also induced dose-dependent repression of Cyp17a1, concomitant with upregulation of Nr0b2, encoding the transcriptional repressor Small Heterodimer Partner (SHP). Overall, the data support a redox-dependent mechanism whereby elevated ROS promotes Nr0b2 expression, leading to Cyp17a1 suppression and impaired androstenedione production. These findings challenge the view of anthocyanins as uniformly beneficial for male fertility and identify Callistemon citrinus as a sustainable source of bioactive anthocyanins capable of modulating redox–endocrine homeostasis in a dose-dependent manner under basal conditions. Full article

Four sweet cherry cultivars (FuChen, Redlight, Huangmi, and Samituo) grown in northern China were used to produce sweet cherry wines with two alcohol levels. Physicochemical properties, antioxidant capacity, and volatile aroma compounds of the wines were systematically investigated. The results showed that wine from the Redlight cultivar with an alcohol content of 11.22 ± 0.17% contained the highest phenolic content and also exhibited the strongest antioxidant capacity as measured by DPPH and ABTS•⁺ assays. Meanwhile, wine from the FuChen cultivar with an alcohol content of 11.45 ± 0.03% had the highest anthocyanin content and showed the strongest FRAP antioxidant activity. Orthogonal partial least squares discriminant analysis (OPLS-DA) based on electronic nose data clearly distinguished the eight sweet cherry wine samples from different cultivars. A total of 58 volatile compounds were identified by headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME-GC-MS). Both principal component analysis (PCA) and OPLS-DA revealed clear differences among the sweet cherry wines based on their volatile composition. Using variable importance in projection (VIP) scores > 1 and relative odor activity values (ROAVs), the key aroma compounds contributing to the characteristic aroma profiles of the eight sweet cherry wines were identified as ethyl butanoate, isoamyl acetate, isoamyl hexanoate, methyl decanoate, ethyl decanoate, ethyl benzoate, methyl salicylate, citronellol, and eugenol. These findings provide important guidance for the selection of raw materials to improve the production of sweet cherry wines with targeted alcohol levels. Full article