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Abstract In this paper, we explored the influence of lipase on cuticle wax and anthocyanin metabolism in blueberries. The results demonstrated a progressive alteration in wax crystal structure, transitions from rod‐like to short rod‐like and flaky, and even disappearance of the cuticle wax. Gas chromatography–mass spectrometry analysis revealed a reduction in esters, aldehydes, long‐chain alkanes, and total wax content following lipase treatment of blueberry fruits. Additionally, ultra high performance liquid chromatography–mass spectrometry analysis showed significant decreases in peonidin, petunidin, cyanidin, and delphinidin, accompanied by an increase in malvidin content due to lipase treatment. Subsequent investigations uncovered that lipase treatment inhibited the activities of key anthocyanin synthetic enzymes, namely phenylalanine ammonia‐lyase (PAL), chalcone isomerase (CHI), and dihydroflavonol reductase (DFR). Moreover, lipase treatment downregulated the expression of synthesis‐related genes, including PAL, 4‐coumarate: CoA ligase, chalcone synthase (CHS), CHI, and DFR. In summary, our findings indicate that lipase disrupts the integrity of cuticle wax structure, inhibits anthocyanin synthesis, and reduces anthocyanin content. Accordingly, our studies suggest that inhibiting lipase could offer a novel approach to preserving the freshness and anthocyanin concentration of blueberries.

Introduction: The scarcity of naturally available sources for blue colorants has driven reliance on synthetic alternatives. Nevertheless, growing health concerns have prompted the development of naturally derived blue colorants, which remains challenging with limited success thus far. Anthocyanins (ACNs) are known for providing blue colors in plants, and metal complexation with acylated ACNs remains the primary strategy to generate stable blue hues. However, this approach can be costly and raise concerns regarding potential metal consumption risks., Objectives: Our study aims to introduce a metal-free approach to achieve blue coloration in commonly distributed non-acylated 3-glucoside ACNs by exploring their interactions with proteins and unveiling the underlying mechanisms., Methods: Using human serum albumin (HSA) as a model protein, we investigated the structural influences of ACNs on their blue color generation using visible absorption spectroscopy, fluorescence quenching, and molecular simulations. Additionally, we examined the bluing effects of six proteins derived from milk and egg and identified the remarkable roles of bovine serum albumin (BSA) and lysozyme (LYS)., Results: Our findings highlighted the importance of two or more hydroxyl or methoxyl substituents in the B-ring of ACNs for generating blue colors. Cyanidin-, delphinidin- and petunidin-3-glucoside, featuring two neighboring hydroxyl groups in the B-ring, exhibited blue coloration when interacting with HSA or LYS, driven primarily by favorable enthalpy changes. In contrast, malvidin-3-glucoside, with two methoxyl substituents, achieved blue coloration through interactions with HSA or BSA, where entropy change played significant roles., Conclusion: Our work, for the first time, demonstrates the remarkable capability of widely distributed 3-glucoside ACNs to generate diverse blue shades through interactions with certain proteins. This offers a promising and straightforward strategy for the production of ACN-based blue colorants, stimulating further research in this field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: Drying is a critical post-harvest process for medicinal plants, which are typically high in moisture and microorganisms. To prevent spoilage and quality loss, it is essential to dry these plants promptly. The drying method significantly impacts the levels of secondary metabolites and the organoleptic characteristics of medicinal plants. This study aimed to investigate the effects of various drying methods on the total phenolics, flavonoids, anthocyanins, antioxidant activity, and phenolic acids in caper (Capparis spinosa L.) fruits. The experiment was performed using a completely randomized design with three replications and included 11 treatments: shade drying, sun drying, oven drying (at 50 °C, 60 °C, and 70 °C), microwave drying (at 300 W, 600 W, and 900 W), freeze-drying, salt-drying, and a fresh plant sample as a control., Results: Among the drying methods tested, microwave drying consistently produced the highest levels of flavonoids, anthocyanins, and antioxidant activity, regardless of wattage. Specifically, the highest total phenol content was observed in samples dried at 900 W microwave, 600 W microwave, and 70 °C in the oven (5.3, 5.37, and 5.31 mg GAE/g DW, respectively). Drying at 600 W microwave yielded the highest levels of caffeic, cinnamic, ferulic, vanillic, and protocatechuic acids (13.03, 3.85, 4.28, 9.73, and 5.6 µg/g, respectively) while drying at 900 W microwave also resulted in elevated levels of caffeic, ferulic, protocatechuic, and p-coumaric acids. The 70 °C oven drying method also showed high levels of caffeic, cinnamic, ferulic, vanillic, and protocatechuic acids. Freeze-drying achieved the highest levels of rosmarinic, gallic, and m-coumaric acids (320.17, 175.3, and 12.99 µg/g, respectively), while shade drying produced high levels of p-hydroxybenzoic, cinnamic, ferulic, m-coumaric, protocatechuic, and p-coumaric acids., Conclusions: Overall, microwave drying (especially at 600 W), oven drying at 70 °C, and freeze-drying emerged as effective alternatives to traditional drying methods. These methods not only preserved the color, texture, and taste of the fruits but also enhanced their bioactive compound levels., Competing Interests: Declarations. Ethics approval and consent to participate: The plant materials utilized in this study were C. spinosa fruits collected from their natural habitats in Khorramabad, Iran. Plant material was sampled on private land with the owner’s permission. We adhered to all relevant institutional, national, and international guidelines and legislation during the collection process. All procedures were carried out accordnig to the guidelines. Consent for publication: Not applicable. Guideline statement: The authors confirm that all methods were carried out following relevant guidelines and regulations. Clinical trial number: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Red Actinidia arguta has recently become highly popular because of its red appearance resulting from anthocyanin accumulation, and has gradually become an important breeding direction. However, regulators involved in anthocyanin biosynthesis have not been fully characterized in A. arguta., Results: Here, we demonstrated that a key R2R3-MYB transcription factor, AaMYB61-like, plays a crucial role in A. arguta anthocyanin biosynthesis. The RT-qPCR results revealed that transient overexpression of AaMYB61-like in A. arguta fruit at 90-100 DAFB significantly promoted anthocyanin biosynthesis, as did the gene expression levels of AaCHS, AaCHI, AaF3H, AaLDOX, and AaF3GT, whereas the result of VIGS revealed the opposite results in A. arguta fruit at 105-115 DAFB. A transcriptional activation assay indicated that AaMYB61-like exhibited transcriptional activation activity. Y1H and LUC assays revealed that AaMYB61-like activates the promoters of AaCHS, AaLDOX, and AaF3GT. In addition, AabHLH137 was found to be related to fruit color from the transcriptome data. We demonstrated that AaMYB61-like promotes anthocyanin biosynthesis by interacting with AabHLH137 via Y2H, BiFC, and Agrobacterium-mediated co-transformation., Conclusions: Our study not only reveals the functions of AaMYB61-like and AabHLH137 in anthocyanin regulation, but also broadly enriches color regulation theory, establishing a foundation for clarifying the molecular mechanism of fruit coloration in kiwifruit., Competing Interests: Declarations. Ethics approval and consent to participate: This study did not include human or animal subjects. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Gloriosa L. possesses exceptional ornamental value, with its floral hues exhibiting a wide range of variations. In this study, we employed sophisticated colorimetry, Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS), and transcriptome sequencing to investigate the phenotypic expression of tepal colors, the composition of carotenoids and anthocyanins, and the differential gene expression in four Gloriosa varieties during their full bloom phase. Our findings revealed that the redness of the tepals, indicated by higher a * values, increased with the intensity of the red hue, while lighter colors corresponded to higher L * values. Metabolomic analysis identified 50 carotenoids and 60 anthocyanins. It was observed that carotenoids primarily influence the yellow and orange color of Gloriosa tepals, with β-carotene, lutein, and zeaxanthin being the predominant carotenoids. Anthocyanins serve as the principal coloring agents in the orange, red and purple tepals of Gloriosa. High levels and proportions of cyanidins and pelargonidins are key contributors to the formation of red and purple tepals, while high levels and proportions of peonidins also play a significant role in purple coloration. In contrast, the presence of high levels and proportions of pelargonidins alone is a crucial factor in the formation of orange tepals. Transcriptomic data unearthed 57 and 92 candidate differentially expressed genes (DEGs) belong to carotenoid and anthocyanin biosynthesis pathway, respectively, with PSY, PDS, DFR, and ANS genes considered as critical genes for the differential accumulation of pigments of Gloriosa tepals. Weighted gene co-expression network analysis (WGCNA) revealed significant co-expression patterns between 217 transcription regulatory factors (TFs) and 8 carotenoid biosynthesis genes, and between 194 TFs and 41 anthocyanin biosynthesis genes. qRT-PCR verified the expression patterns of four carotenoid biosynthesis-related genes, eight anthocyanin biosynthesis-related genes, and three transcription regulatory factors. It was found that Cluster-121969.6 (MYB) gene is specifically expressed in the tepals of the four varieties (compared to stems and leaves) and shows a high consistency with the trend of anthocyanin content changes. This research provides new insights into the mechanisms underlying the formation of diverse floral colors in Gloriosa tepals., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Anthocyanin is an important class of water-soluble pigments that are widely distributed in various tissues of plants, and it not only facilitates diverse color changes but also plays important roles in various biological processes. Maize silk, serving as an important reproductive organ and displaying a diverse range of colors, plays an indispensable role in biotic resistance through its possession of anthocyanin. However, the copy numbers, characteristics, and expression patterns of genes involved in maize anthocyanin biosynthesis are not fully understood. In this study, gene numbers, distribution, structure, cis-elements of the anthocyanin synthetic gene family were identified, and then the potential transcriptional factors were predicted by two analyzed methods. Finally, genes involved in maize silk pigment were screened by un-targeted metabolism analysis., Results: Ten gene families involved in the maize anthocyanin biosynthesis pathway were identified, and 142 synthetic genes were obtained. These anthocyanin biosynthetic genes have high copy numbers and are normally clustered on chromosomes. The promoters of these synthetic genes contain various cis-elements and the gene expression patterns and transcriptional regulatory networks were analyzed. These genes are distributed on different chromosomes and gene expression patterns vary across different tissues in maize. Specifically, these genes often exhibit higher expression in the stem, leaves, and seeds. Ten highly expressed genes in silks were identified. Based on un-targeted metabolites detection in the silks of four maize representative inbred lines with different colors, two main differential anthocyanin components were identified. Furthermore, the gene expression patterns of the ten highly expressed genes and their potential interacting transcriptional factors were analyzed across the four inbred lines., Conclusions: The results in this study show a through picture of maize anthocyanin synthetic genes, and the structure and function of genes related to anthocyanin biosynthesis in maize could be further investigated., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests. Clinical trial number: Not applicable., (© 2025. The Author(s).)

Background: Wheat seeds display different colors due to the types and contents of anthocyanins, which is closely related to anthocyanin metabolism. In this study, a transcriptomic and metabolomic analysis between white and purple color wheat pericarp aimed to explore some key genes and metabolites involved in anthocyanin metabolism., Results: Two wheat cultivars, a white seed cultivar Shiluan02-1 and purple seed cultivar Hengzi151 were used to identify the variations in differentially expressed genes (DEGs) and differentially accumulated flavonoids (DAFs). Based on metabolomic data, 314 metabolites and 191 DAFs were identified. Chalcone, flavonol, pro-anthocyanidin and anthocyanidin were the most differentially accumulated flavonoid compounds in Hengzi151. 2610 up-regulated and 2668 down-regulated DEGs were identified according to transcriptomic data. The results showed that some structural genes in anthocyanin synthesis pathway were prominently activated in Hengzi151, such as PAL, CAD, CHS and so on. Transcription factors (TFs) of MYB, bHLH, WD40 and some other TFs probably involved in regulating anthocyanin biosynthesis were identified. Some genes from hormone synthetic and signaling pathways that may participate in regulating anthocyanin biosynthesis also have been identified., Conclusions: Our results provide valuable information on the candidate genes and metabolites involved in the anthocyanin metabolism in wheat pericarp., Competing Interests: Declarations. Ethics approval and consent to participate: All experimental research and field studies on plants in our study complies with Chinese institutional, national, and international guidelines and legislation. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: The quality of soybeans is reflected in the seed coat color, which indicates soybean quality and commercial value. Researchers have identified genes related to seed coat color in various plants. However, research on the regulation of genes related to seed coat color in soybeans is rare., Methods: In this study, four lines of seed coats with different colors (medium yellow 14, black, green, and brown) were selected from the F 2:5 population, with Beinong 108 as the female parent and green bean as the male parent, and the dynamic changes in the anthocyanins in the seed coat were stained with 4-dimethylaminocinnamaldehyde (DMACA) during the grain maturation process (20 days from grain drum to seed harvest). Through RNA-seq of soybean lines with four different colored seed coats at 30 and 50 days after seeding, we can further understand the key pathways and gene regulation modules between soybean seed coats of different colors., Results: DMACA revealed that black seed coat soybeans produce anthocyanins first and have the deepest staining. Clustering and principal component analysis (PCA) of the RNA-seq data divided the eight samples into two groups, resulting in 16,456 DEGs, including 5359 TFs. GO and KEGG enrichment analyses revealed that the flavonoid biosynthesis, starch and sucrose metabolism, carotenoid biosynthesis, and circadian rhythm pathways were significantly enriched. We also conducted statistical and expression pattern analyses on the differentially expressed transcription factors. Based on weighted gene coexpression network analysis (WGCNA), we identified seven specific modules that were significantly related to the four soybean lines with different seed coat colors. The connectivity and functional annotation of genes within the modules were calculated, and 21 candidate genes related to soybean seed coat color were identified, including six transcription factor (TF) genes and three flavonoid pathway genes., Conclusions: These findings provide a theoretical basis for an in-depth understanding of the molecular mechanisms underlying differences in soybean seed coat color and provide new genetic resources.

Chilling stress seriously affects the growth and yield of tomato. Anthocyanin is a typical chilling-induced metabolite with strong antioxidant activity and photoprotective capacity. Here, we found that anthocyanin was also involved in ascorbic acid biosynthesis and glycolysis under chilling stress. SlAN2 is an important positive gene in anthocyanin biosynthesis. The results of physiological indicators showed that SlAN2 overexpression lines (A189) had a greater ability to tolerate cold stress than wild-type (WT) plants. Conjoint analysis of transcriptomics and metabonomics of A189 lines and WT plants was used to analyze the metabolic difference and the cold resistance mechanisms caused by anthocyanin under chilling stress. The anthocyanin accumulated more in A189 than that in WT under chilling stress at 4 °C for 24 h, which led to hexoses and ascorbic acid increased significantly. Results indicate that SlAN2 overexpression reduces the expression of key enzyme genes in glycolytic pathway such as phosphofructokinase (PFK) and pyruvate kinase (PK) genes, weakens glycolysis ability, and promotes accumulation of hexoses in A189 lines at 4 °C for 24 h compared with wild lines. Additionally, ascorbic acid content is increased by up-regulated the genes of ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR). The increased hexose content can reduce cell osmotic potential, freezing point and synthesize more ascorbic acid, while the increased ascorbic acid content can enhance the ability to scavenge reactive oxygen species, so improves the cold resistance of tomato. The glycolysis and ascorbic acid metabolism pathway mediated by SlAN2 provides a new insight for the molecular mechanism of anthocyanins in improving the cold resistance of tomato and provides a new theoretical basis for cultivating new cold-tolerant tomato varieties., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Crabapple is a valuable tree species in gardens due to its captivating array of flower and leaf colors, rendering it a favored choice in landscaping. The economic and ornamental values of Malus crabapple are closely associated with the biosynthesis of anthocyanin, a pigment responsible for its vibrant hues. The intricate regulation of anthocyanin biosynthesis involves the concerted activity of various genes. However, the specific mechanism governing this process in crabapple warrants in-depth exploration. In this study, we explored the inhibitory role of MsMYB62-like in anthocyanin biosynthesis. We identified MsDFR and MsANS as two downstream target genes of MsMYB62-like. These genes encode enzymes integral to the anthocyanin biosynthetic pathway. The findings demonstrate that MsMYB62-like directly binds to the promoters of MsDFR and MsANS , resulting in the downregulation of their expression levels. Additionally, our observations indicate that the plant hormone cytokinins exert a suppressive effect on the expression levels of MsMYB62-like , while concurrently upregulating MsDFR and MsANS . This study reveals that the MsMYB62-like- MsDFR / MsANS module plays an important role in governing anthocyanin levels in Malus crabapple. Notably, the regulatory interplay is modulated by the plant hormone cytokinins.

Fruit ripening is a highly-orchestrated process that requires the fine-tuning and precise control of gene expression, which is mainly governed by phytohormones, epigenetic modifiers, and transcription factors. How these intrinsic regulators coordinately modulate the ripening remains elusive. Here we report the identification and characterization of FvALKBH10B as an N 6 -methyladenosine (m 6 A) RNA demethylase necessary for the normal ripening of strawberry (Fragaria vesca) fruit. FvALKBH10B is induced by phytohormone abscisic acid (ABA), and ABA-Responsive Element Binding Factor 3 (FvABF3), a master regulator in ABA signaling, is responsible for this activation. FvALKBH10B mutation leads to a delay in fruit ripening and causes global m 6 A hypermethylation of 1859 genes. Further analyses show that FvALKBH10B positively modulates the mRNA stability of SEPALLATA3 (FvSEP3) encoding a transcription factor via m 6 A demethylation. In turn, FvSEP3 targets numerous ripening-related genes including those associated with biosynthesis of ABA and anthocyanin and regulates their expression. Our findings uncover an FvABF3-FvALKBH10B-FvSEP3 cascade in controlling fruit ripening in strawberry and provide insights into the complex regulatory networks involved in this process., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Cinnamomum camphora, a key multifunctional tree species, primarily serves in landscaping. Leaf color is crucial for its ornamental appeal, undergoing a transformation to red that enhances the ornamental value of C. camphora. However, the molecular mechanisms underlying this transformation remain largely unexplored. In this study, green leaf (GL), color turning red leaf (RL) and whole red leaf (WRL) were obtained to measure pigment contents, while GL and RL were analyzed for transcriptomic alterations. A decline in chlorophyll content and a rise in anthocyanins were observed during the transition from green to red leaves. Using LC MS/MS, 11 types of anthocyanins showed significant accumulative differences, with cyanidin-3,5-O-diglucoside exhibiting the greatest disparity. Comparative RNA-seq identified 22,948 genes against reference genes, revealing 544 novel genes. Of these, 3,222 genes were up-regulated and 7,391 genes were down-regulated when the FPKM mean value > 1 in at least one group. The ribosome was identified as the most abundant KEGG term, with a substantial number of down-regulated differentially expressed genes (DEGs). The results indicated a downward trend in protein content, with GL exhibiting the highest protein concentration. 22, 4, and 29 DEGs were associated with chlorophyll biosynthesis, chlorophyll degradation, and anthocyanin biosynthesis, respectively. Most DEGs related to chlorophyll biosynthesis were down-regulated. SGR and SGRL, which are associated with chlorophyll degradation, exhibited opposite differential expression, resulting in a significant decrease in chlorophyll content in RL. The significantly up-regulated genes ANS and UFGT are advantageous for anthocyanin biosynthesis, contributing to the red coloration observed. Additionally, differential expression was noted in 40 R2R3-MYBs. Two MYB90 (Ccam01G003512 and Ccam01G003515) homologs of AtMYB113 were also identified showed high levels of up-regulation in RL. These findings suggest a strong correlation between pigment metabolism and transcriptome data, elucidating the mechanism that leads to the red coloration of leaves in C. camphora., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Rapeseed (Brassica napus L.) possesses substantial economic value as an oil, vegetable, and forage crop, while also exhibiting notable ornamental characteristics. Recent advances in flower colour breeding have significantly enhanced the visual appeal of rapeseed, with anthocyanins identified as the primary contributor to the development of red, purple, and pink flowers. However, the mechanisms underlying the synthesis and regulation of anthocyanins during petal coloration in rapeseed are still poorly understood. This research combined miRNA and mRNA expression data from four different color phases, along with degradome analysis, to discover important miRNA-mRNA modules responsible for controlling the accumulation of anthocyanin in purple-flowered rapeseed. In the process of petal development, a grand sum of 247 miRNAs (including 223 known and 24 novel miRNAs) were effectively detected, with 64 of them displaying differential expression patterns. Degradome sequencing was used to conduct a comprehensive analysis of 152 targets for the differential expression miRNAs. Out of these, 108 miRNA-mRNA modules exhibit contrasting expression patterns. Some miRNAs and their corresponding targets have additionally been discovered, potentially playing a role in governing the buildup of anthocyanin in purple-flowered rapeseed. The regulatory modules miR156-SPL9 and miR828-PAP2 composed of miR156b and miR828 and their targets may play a key role in this process. The results offer a thorough analysis of miRNAs linked to the regulation of anthocyanin in B. napus, offering valuable understanding into the regulatory processes that govern miRNA-mediated anthocyanin production in Brassica crops., Competing Interests: Declarations. Ethics approval and consent to participate: This study including the collection on plants material complies with relevant institutional, national, and international guidelines and legislation. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The color variation of the leaves in autumn is a significant ornamental feature of Acer truncatum Bunge, especially when the leaves gradually become redder. Many studies focused on leaf color changes; however, less research has been conducted on the mechanism by which A. truncatum 's autumn leaves turn red. Red, middle and green leaves of Acer truncatum were used as the study materials to evaluate their flavonoid-related metabolites and infer gene and metabolite expression patterns in conjunction with transcriptome expression. For a start, phenotypic and leaf color parameters analyses showed that red leaves had the highest color redness and greenness (a*). In addition, a total of 23 flavonoid-related metabolites were identified through the metabolome, including five anthocyanins. Of them, cyanidin 3-O-β-D-sambubioside, cyanidin 3-O rutinoside, pelargonidin 3-O-3″,6″-O-dimalonylglucoside, delphinidin 3,7-di-O-β-D-glucoside and 3-O-β-D-sambubioside would help the leaves turn red in A. truncatum . Similarly, combined transcriptomics and metabolomics analyses showed that most genes in the flavonoid and anthocyanin biosynthetic pathways were differentially expressed in both types of leaves. Chalcone synthase ( CHS ), dihydroflavonol 4-reductase ( DFR ) and anthocyanin synthase ( ANS ) could affect flavonoid synthesis during leaf color change. This study could provide data for the genetic improvement of maple plants by exploring valuable metabolites and genes in flavonoid synthesis, and enhance the understanding of different developmental stages.

Background: The anthocyanin synthase (ANS) gene is pivotal in plant anthocyanin biosynthesis, significantly influencing fruit coloration. Despite extensive on various studies, there remains a notable absence of research on the regulatory functions of the ANS gene in the 'Lingwuchangzao' variety under elevated temperature and drought stress., Results: In this study, 107 ZjANS genes were identified. Analyses of their physicochemical properties and gene structures revealed that most are localized in the cytosol and nucleus, with only seven ZjANS genes exhibiting basic characteristics. All genes contained the conserved domains 2OG-FeII_Oxy and DIOX-N. Chromosomal localization and Ka/Ks analysis indicated that these genes are distributed across 12 chromosomes, with 22 tandem gene pairs, suggesting purifying selection. Phylogenetic and synteny analysis demonstrated that ZjANS genes are evolutionarily conserved, forming collinear gene pairs with four species. Cis-acting element analysis identified elements related to anthocyanin synthesis and environmental stress, such as LTR and MBS. Predictions of protein interaction indicated that certain ZjANS proteins interact with MYB, CHI, and GA proteins, potentially involved in the anthocyanin synthesis pathway. Transcriptome and qRT-PCR analysis revealed that thirteen ZjANS genes positively respond to coloring in jujube under elevated temperature and drought stress, with ZjANS17 identified as particularly significant., Conclusion: This study identified 107 ZjANS genes and systematically analyzed their evolutionary conservation. Thirteen ZjANS genes were found to positively influence fruit coloring in 'Lingwuchangzao' under elevated temperature and drought stress. These findings provide a foundation for further investigations into ZjANS genes and the breeding new jujube varieties., Competing Interests: Declarations. Ethics approval and consent to participate: The plant material of this study protocol comply with relevant institutional, national, and international guidelines and legislation. All experimental protocols were approved by the Ningxia University. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The basic helix-loop-helix (bHLH) family is one of the three major transcription factor families that play important transcriptional regulatory roles in plant growth and development. One of the most crucial elements in defining Lagerstroemia indica's decorative qualities is flower color. However, the function of the bHLH transcription factor family in L. indica anthocyanin glycoside synthesis has not been clarified. Using the transcriptome data of flower color, 79 LibHLH genes were found in this study. Phylogenetic analysis showed that the LibHLH genes can be divided into 16 subfamilies, and most of the genes in the same subfamily have similar conserved motifs. The total anthocyanin glycoside content of L. indica 'Zihua Guifei' petals was determined during three developmental stages of the petals' growth. The results showed that the total anthocyanin glycoside content grew gradually with growth and development, and that it accumulated most during the full bloom stage. By using gene expression analysis, protein interaction network analysis, and bioinformatics, it was possible to determine which member of the III f family, LibHLH29, is important for the synthesis of anthocyanin glycosides in L. indica. Its expression was confirmed by qRT-PCR, and the results were essentially compatible with the transcriptome data. It was more prominent in the light-colored bloom stage the color-transition stage of L. indica 'Zihua Guifei'. It can be further investigated as a major candidate gene for regulating anthocyanin glycoside synthesis in L. indica, thus laying the foundation for an in-depth study of the interactions among transcription factors., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Cadmium (Cd) is a toxic heavy metal that interferes with essential metabolic pathways crucial for plant growth, often resulting in toxicity and plant death. Blueberry plants exhibit metabolic adaptations to mitigate the stress caused by elevated Cd levels. In this review, we highlighted the effects of Cd-induced stress on blueberry plants and explored the potential alleviating effects of arbuscular mycorrhizal fungi (AMF). Cd uptake disrupts plant metabolism and impacts primary and secondary metabolites, including anthocyanins, which play a role in defense mechanisms against pathogens. Hence, Cd-induced stress alters anthocyanin levels in blueberry leaves, negatively affecting antioxidant defense mechanisms and hindering growth. Conversely, AMF establishes a symbiotic relationship with blueberry plants, promoting nutrient absorption and enhancing stress tolerance. Understanding the association between Cd stress, anthocyanin responses in blueberries, and AMF-mediated mitigation is crucial for developing integrated strategies to enhance blueberry plant health and improve quality. Employing AMF to remediate metal-related stress represents a significant breakthrough for sustainable crop production in a Cd-contaminated environment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

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., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Research involving human participants and, or animals: Not applicable. Clinical Trial Study: Not applicable. Clinical trial number: Not applicable. Informed consent: Not applicable. Statement specifying permissions: For this study, we acquired permission to study Berberis crataegina issued by the Agriculture and Forestry Ministry of the Republic of Türkiye. Statement on experimental research and field studies on plants: The either cultivated or wild-growing plants sampled comply with relevant institutional, national, and international guidelines and domestic legislation of Türkiye. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Blue barley grain pigmentation results from anthocyanin accumulation in the aleurone layer. Anthocyanins are known for their beneficial effects on human health. The gene encoding the MYELOCYTOMATOSIS 2 (MYC2) transcription factor is potentially responsible for the blue coloration of the aleurone. In non-pigmented barley, a single nucleotide insertion in this gene causes a frameshift mutation with a premature stop codon. It was hypothesized that restoring the MYC2 reading frame could activate anthocyanin accumulation in the aleurone. Using a targeted mutagenesis approach in the present study, the reading frame of MYC2 was restored in the non-pigmented cultivar Golden Promise. Genetic constructs harboring cas9 and gRNA expression units were developed, pre-validated in protoplasts, and then functional MYC2 alleles were generated at the plant level via Agrobacterium -mediated transformation. Anthocyanin accumulation in the aleurone layer of grains from these mutants was confirmed through microscopy and chemical analysis. The expression of anthocyanin biosynthesis genes was analyzed, revealing that the restoration of MYC2 led to increased transcript levels of F3H and ANS genes. These results confirm the critical role of the MYC2 transcription factor in the blue aleurone trait and provide a biotechnological solution for enriching barley grain with anthocyanins.

Background : Verbena stricta is a perennial herb of the Verbenaceae family, known for its medicinal properties, wide adaptability, and high resistance. Methods : This research investigated the metabolic pathways of flower color change by combining transcriptome and metabolomics analyses. Results : In purple flowers and white variants, a total of 118 differentially accumulated metabolites (DAMs), including 20 anthocyanins, and 7627 differentially expressed genes (DEGs) were found. The downregulation of delphinidin-3-O-galactoside, delphinidin-3-O-glucoside, and delphinidin-3-O-(6″-O-p-coumaroyl) glucoside, along with the absence of petunidin and malvidin derivatives, may explain the loss of pigmentation in the white-flower mutant. Fourteen candidate genes involved in anthocyanin biosynthesis were identified, among which the expression of Flavonoid 3', 5'-hydroxylase (F3'5'H) was significantly downregulated, notably limiting flux through the delphinidin pathway and reducing delphinidin accumulation. This limitation in upstream reactions, coupled with the multi-shunt process in downstream reactions, completely blocked the production of petunidin and malvidin. Conclusions : These findings offer new opinions on the anthocyanin metabolites and key genes responsible for the floral pigmentation in V. stricta . Additionally, the white variant provides a valuable platform for future research into the ornamental flower color of the Verbenaceae family., Competing Interests: The authors declare no conflicts of interest.

WD40 proteins play important roles in the synthesis and regulation of anthocyanin, the regulation of plant morphology and development, and the response to various abiotic stresses. However, the role of WD40 in Fragaria vesca ( F. vesca ) has not been studied. In this study, a total of 216 FvWD40 family members were identified, which were divided into four subfamilies based on evolutionary tree analysis. Subcellular localization predictions show that FvWD40 family members are mainly localized in chloroplasts, nuclei, and cytoplasm. An analysis of collinearity revealed a total of eight pairs of intraspecific collinearity of the FvWD40 gene family, and interspecific collinearity showed that the FvWD40 gene family covaried more gene pairs with Arabidopsis thaliana ( Arabidopsis ) than with rice ( Oryza sativa ). Promoter cis-acting elements revealed that the FvWD40 gene family contains predominantly light, hormone, and abiotic stress response elements. Tissue-specific expression analysis showed that a number of members including FvWD40-111 and FvWD40-137 were highly expressed in all tissues, and a number or members including FvWD40-97 and FvWD40-102 were lowly expressed in all tissues. The FvWD40 gene family was found to be expressed at all four different coloring stages of F. vesca by qRT-PCR, with lower expression at the 50% coloring stage (S3). FvWD40-24 , FvWD40-50 , and FvWD40-60 showed the highest expression during the white fruit stage (S1) period, suggesting that these genes play a potential regulatory role in the pre-fruit coloring stage. FvWD40-62 , FvWD40-88 and FvWD40-103 had the highest expression at the 20% coloration stage (S2), and FvWD40-115 , FvWD40-170 , FvWD40-184 and FvWD40-195 had the highest expression at the full coloration stage (S4). These results suggest a potential role for these genes during fruit coloration. This study lays a foundation for further research on the function of the WD40 gene family.

The herbicide fluridone (1-methyl-3-phenyl-5-[3-trifluoromethyl (phenyl)]-4(1H)-pyridone) interferes with carotenoid biosynthesis in plants by inhibiting the conversion of phytoene to phytofluene. Fluridone also indirectly inhibits the biosynthesis of abscisic acid and strigolactones, and therefore, our study indirectly addresses the effect of reduced ABA on the roots and leaf buds development in stem cuttings of Salix babylonica L. 'Tortuosa'. The stem cuttings were kept in distilled water (control) or in a solution of fluridone (10 mg/L) in natural greenhouse light and temperature conditions. During the experiments, morphological observations were carried out on developing roots and leaf buds, as well as their appearance and growth. After three weeks of continuous treatments, adventitious roots and leaf buds were collected and analysed. Identification and analysis of anthocyanins were carried out using micro-HPLC-MS/MS-TOF, while HPLC-MS/MS was used to analyse phenolic acids, flavonoids and salicinoids. The fluridone applied significantly inhibited root growth, but the number or density of roots was higher compared to the control. Contents of salicortin and salicin were several dozen times higher in leaf buds than in roots of willow. Fluridone increased the content of salicortin in roots and leaf buds and declined the level of salicin in buds. Fluridone also declined the content of most anthocyanins in roots but enhanced their content in buds, especially cyanidin glucoside, cyanidin galactoside and cyanidin rutinoside. Besides, fluridone markedly decreased the level of chlorophylls and carotenoids in the leaf buds. The results indicate that applied fluridone solution reduced root growth, caused bleaching of leaf buds, and markedly affected the content of secondary metabolites in the adventitious roots and leaf buds of S. babylonica stem cuttings. The paper presents and discusses in detail the significance of fluridone's effects on physiological processes and secondary metabolism.

The postharvest lighting environment is a main factor that influences quality preservation for harvested biomass. The objective of this study was to evaluate postharvest changes in bioactive compounds of lettuce with different storage light spectra. The effects of green LEDs with peaks at 500 nm and 530 nm, white LEDs (400-700 nm), and dark storage were evaluated, where light intensity (10 μmol m-2 s-1) and photoperiod (12 h per day) were constant with air temperature at 5°C over the 14 d treatment period. Lettuce stored with 500 nm and 530 nm green LEDs exhibited 1474.5% and 1451.8% (approximately 15.7 and 15.5 times) higher antioxidant activity, respectively, compared to dark storage. Significant improvements in total phenolic content, and 67.5% and 64.8% increases in total soluble solids with 530 nm and 500 nm green LEDs over dark storage were discerned. Exposure to 530 nm green LEDs led to 128.2% (approximately 2.28 times) higher anthocyanin content, a 26.2% increase in carotenoids, and a 95% rise in flavonoid content compared to dark storage. Increases of 26.4% and 16.0% in chlorophyll a content in lettuce stored under 500 nm and 530 nm green LEDs, respectively, and 65.6% and 46.6% rises in the Chlorophyll a/b ratio were observed. Compared to dark storage, green LEDs (500 nm) resulted in a 13.5% higher total chlorophyll content. Findings underscore the positive impact of green LEDs on the nutritional quality of lettuce, providing insight for postharvest practices., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Salehinia et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Aeonium arboreum 'Halloween', a popular indoor ornamental succulent in China, changes its leaf colour to red on light exposure. However, the underlying molecular mechanisms is still vague. Comparative analysis of transcriptome data from 'Halloween' leaves treated under dark and light conditions revealed two R2R3-MYB transcription factors, AaMYB113 and AaMYB114, that may mediate anthocyanin accumulation. In this study, we cloned the AaMYB113 and AaMYB114 genes, encoding proteins of 279 and 248 amino acids, respectively. Transcriptional activity analysis revealed that AaMYB113 exhibits strong transcriptional activity, in contrast to AaMYB114, which demonstrates minimal activity. Transient expression studies in tobacco leaves demonstrated that AaMYB113 induced red pigmentation, whereas AaMYB114 did not. Subsequent stable overexpression in Arabidopsis thaliana confirmed that AaMYB113, but not AaMYB114, could similarly turn Arabidopsis leaves red. Further stable transformation of AaMYB113 in tobacco affected multiple floral components, including leaves, petals, calyx, flower tubes, and filaments, turning them red. Quantitative real-time PCR (qRT-PCR) assay in leaves of AaMYB113 stably transformed tobacco and Arabidopsis revealed upregulation of anthocyanin biosynthesis-related structural genes and TT8-like transcription factors. Moreover, the dual luciferase analysis confirmed that AaMYB113 can activate the promoters of 'Halloween' anthocyanin synthesis structural genes, AaCHS, AaCHI, AaF3H, AaDFR and AaANS. The above results indicate that AaMYB113 can promote anthocyanin synthesis, while AaMYB114 does not have this function. This study contributes significantly to the limited body of research on the molecular mechanisms of anthocyanin synthesis in succulents, advancing our understanding of how these pathways are regulated in 'Halloween' succulents and potentially other species., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Tea is an important cash crop that significantly contributes to rural development, poverty reduction and food security in many developing countries. It provides livelihoods for millions of smallholder producers and aids their economic stability. Anthocyanins in tea leaves provides excellent commercial quality and germplasm exploration potential. These compounds give tea leaves vibrant colors and increase health benefits. The current understanding of the synergistic regulation mechanisms responsible for color changes in purple tea, attributed to anthocyanin degradation, remains unclear., Results: In this study, we have identified 30 gene families within the genome that are associated to with anthocyanin metabolism from tea. These gene families play distinct roles in the biosynthesis of anthocyanin including the formation of the core, structure, modification of the molecular framework, facilitation of transport process, regulation of gene expression, breakdown pathways, sugar transportation and iron ion respectively. Subsequently, we investigated the synergistic mechanisms of anthocyanin metabolism related gene families within tea leaves using structural equation modeling. The results showed that sugar transport positively affects anthocyanin transportation, and promotes anthocyanin degradation during leaf pigmentation, whereas, it inhibits anthocyanin degradation during the fading of leaf color. Further, Iron ions facilitate the degradation of anthocyanins during their deposition and conversely, impede this degradation process during digestion. These finding suggests that tea plants may regulate the synthesis and degradation of anthocyanins through sugar transport and iron ions ensure healthy levels and vibrant colors., Conclusions: Our study contributes valuable information into the dynamic equilibrium anthocyanin mechanism and sheds light on complex regulatory mechanisms that govern the synthesis, transport and degradation of these pigments. These insights could be further used to develop strategies for enhancing anthocyanins content in unique tea germplasm to aid tea industry in producing new tea products with increased health benefits and aesthetic appeals., Competing Interests: Declarations Ethics approval and consent to participate This study was carried out in compliance with the relevant institutional, national, as well as international guidelines and legislation, and comply with the relevant rules of the Tea Research Institute, Yunnan Academy of Agricultural Sciences. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: In plants, Cytochrome P450 (CYP450) constitutes the largest family of metabolic enzymes and plays a crucial role in various physiological processes, including plant growth and development. Eggplants are well-known for their peel's high concentration of anthocyanin compounds that provide significant health benefits to humans. The accumulation of anthocyanins in eggplant peels is an important process during growth and development. Therefore, it is essential to identify the CYP450 genes in eggplant (SmCYPs) and analyze their expression profiles during the period of anthocyanin accumulation in the peel., Results: A total of 180 SmCYPs were identified in the eggplant genome and classified into eight subfamilies based on phylogenetic analysis. These SmCYPs exhibited highly conserved gene structure (exon/intron) and protein motifs, especially within their respective subgroup. Sixteen pairs of genes with collinearity were identified through gene duplication analysis. Promoter cis-acting element analysis revealed that SmCYPs are involved in various responses, including growth and development, stress responsiveness, and light responsiveness. Transcriptome data analysis revealed that all SmCYPs were expressed in various eggplant tissues, such as roots, stems, leaves, flowers, and fruits; with diverse expression patterns among members. The expression patterns of SmCYPs in eggplant peel also exhibited diversity during different stages of anthocyanin accumulation. qRT-PCR analysis demonstrated similar expression patterns for 15 selected SmCYPs as observed in the transcriptome data. Metabolomics analysis further suggested that SmCYPs are involved in the biosynthesis of secondary metabolites and metabolic pathways related to flavonoid and flavone/flavonol biosynthesis. Notably, three specific SmCYPs (SmCYP73A1/75A/98A1) play a significant role in flavonoid biosynthesis, particularly in anthocyanin synthesis in eggplant., Conclusion: Genome-wide identification, phylogenetic analysis, expression profile analysis, and exploration of metabolic pathways related to SmCYPs provide valuable insights into the roles of these genes in anthocyanin accumulation in various tissues and organs, including eggplant peel. The findings from this study lay a foundation for the functional analysis of SmCYPs involvement in anthocyanin accumulation in eggplant peel, providing a molecular basis for breeders to cultivate novel varieties of eggplants with high levels of anthocyanin content., (© 2024. The Author(s).)

Phenolic acids have health-promoting properties, however, but their low concentrations in Salvia miltiorrhiza limit broader medicinal applications. MYB and bHLH transcription factors activate multiple target genes involved in phenylpropanoid metabolism, thereby enhancing the production of various secondary metabolites. We introduced the MYB transcription factor Antirrhinum Rosea1 ( AmROS1 ) or Delila ( AmDEL ) into S. miltiorrhiza and observed that antioxidant activity in transgenic plants increased by 1.40 to 1.80-fold. The total content was significantly higher in transformants compared to the controls. Furthermore, heterologous expression of AmROS1 or AmDEL triggered moderate accumulations of rosmarinic acid and salvianolic acid at various growth stages. Levels of total phenolics, total flavonoids, and anthocyanins were significantly elevated. These biological and phytochemical alterations were correlated with the upregulated expression of genes involved in phenolic acid biosynthesis. Our findings demonstrate that AmROS1 and AmDEL function as a transcriptional activator in phenolic acids biosynthesis. This study offers further insights into the heterologous or homologous regulation of phenolics production, potentially enabling its engineering in S. miltiorrhiza .

Acetaldehyde is a key carbonyl by-product during red wine alcoholic fermentation; it is reactive and takes part in certain reactions involving anthocyanin. The aim of this study was to investigate the influence of SO 2 and condensed tannin on the acetaldehyde accumulation of Saccharomyces cerevisiae ( S. cerevisiae ) during alcoholic fermentation and the ripple effect on wine anthocyanin. In this study, six sets of Cabernet Sauvignon alcoholic fermentation with two different sulfur levels (HS and LS) were carried out by adding exogenous condensed tannins before fermentation (T0) in the acetaldehyde rise period (TA) of S. cerevisiae and at the end of fermentation (TE), separately. The acetaldehyde evolution was identified during fermentation and anthocyanin was analyzed comparatively. The results showed that HS treatment slowed down the degradation of acetaldehyde, while tannins accelerated the degradation of acetaldehyde during alcoholic fermentation, especially TA wines. Furthermore, TA wines possessed a unique anthocyanin profile after fermentation regardless of SO 2 level compared with other wines. These results suggest that acetaldehyde-mediated anthocyanin polymerization most likely occurs timely at the acetaldehyde production phase of S. cerevisiae during alcoholic fermentation, and managing tannin addition time during production could be used to regulate the anthocyanin profile.

Colored leaves, a notable horticultural trait, have high research and ornamental value. The evergreen sweet olive (Osmanthus fragrans), one of the top ten traditional flowers in China, has been cultivated for more than two thousand years. However, in recent years, an increasing number of O. fragrans cultivars with colored leaves have been cultivated for their ornamental value. To study the molecular mechanism underlying the observed changes in leaf color, we selected O. fragrans 'Yinbi Shuanghui' (Y), which has yellow-white leaves, and O. fragrans 'Sijigui' (S), which has green leaves, as materials. Pigment content measurement showed that the chlorophyll, carotenoid and anthocyanin contents in Y were lower than in S. According to the SWATH-MS sequencing results, a total of 3,959 proteins were quantitatively identified, 1,300 of which were differentially expressed proteins (DEPs), including 782 up-regulated and 518 down-regulated proteins in Y compared to S. Functional enrichment analysis of DEPs revealed that down-regulated expression of photosynthesis related proteins may lead to the inhibition of chlorophyll synthesis in Y, this may be the main cause of leaf color change. Moreover, a protein interaction prediction model also showed that proteins such as PetC, PsbO, PsbP, and PsbQ were key proteins in the interaction network, and the up-regulated proteins participating in the anthocyanin and carotenoid pathways may be related to the formation of yellow-white leaves. Taken together, our findings represent the first SWATH-MS-based proteomic report on colored leaf O. fragrans and reveal that chlorophyll synthesis and secondary metabolism pathways contribute to the changes in leaf color., (© 2024. The Author(s).)

Background: This study examines the effects of various complementary light spectra on the growth, development, antioxidant activity, and nutrient absorption in strawberry plants under stress conditions. Light-emitting diodes (LEDs) were used to provide specific wavelengths, including monochromatic blue (460 nm), monochromatic red (660 nm), a dichromatic mix of blue and red (1:3 ratio), full-spectrum white light (400-700 nm), and ambient light as a control (no LED treatment). The stress treatments applied were: control (no stress), salinity (80 mM NaCl), alkalinity (40 mM NaHCO₃), and a combined salinity/alkalinity condition., Results: Our results indicated that complementary light spectra, especially red and blue/red, helped mitigate the adverse effects of stress on plant growth and development. These spectra improved plant tolerance by enhancing the activity of polyphenol oxidase and peroxidase enzymes and increasing starch accumulation in the leaves. Furthermore, under stress conditions, red and blue-red light significantly boosted fruit anthocyanin levels. Although stress elevated antioxidant activity, supplementary light reduced this activity by alleviating stress compared to ambient light. While stress led to increased Na and Cl ion concentrations in leaves, treatments with blue, red, and blue-red light minimized these harmful effects and promoted the absorption of beneficial ions such as K, Mg, Fe, and Cu., Conclusions: Adjusting light quality significantly influences the morphology and physiology of strawberry plants, underscoring the role of specific light spectra in promoting optimal growth under stress conditions., Clinical Trial Number: Not applicable., (© 2024. The Author(s).)

Background: Strawberries are bright in color, sweet and sour in taste, and rich in nutrients and flavonoid compounds such as anthocyanins and proanthocyanidins. The synthesis and accumulation of anthocyanins are the decisive factors that make strawberries appear bright red. From the perspective of plant breeding, a change in flesh color is an important goal., Methods: In this study, two strawberry plants with different flesh colors were selected, and transcriptome and metabolome analyses were performed during the color change period (S1) and ripening period (S2)., Results: RNA-seq revealed a total of 13,341 differentially expressed genes (DEGs) between and within materials, which were clustered into 5 clusters. A total of 695 metabolites were detected via metabolome analysis, and 243 differentially regulated metabolites (DRMs) were identified. The anthocyanin biosynthesis, starch and sucrose metabolism and glycolysis/gluconeogenesis pathways were determined to be important regulatory pathways for changes in strawberry flesh color through a joint analysis of RNA-seq data and the metabolome. The leucoanthocyanidin reductase (LAR) and chalcone synthase (CHS) gene is a key gene related to anthocyanins, cinnamic acid, and phenylalanine. In addition, through joint RNA-seq and metabolome analyses combined with weighted gene co-expression network analysis (WGCNA), we identified 9 candidate genes related to strawberry flesh color., Conclusions: Our research findings have laid the groundwork for a more comprehensive understanding of the molecular mechanisms governing the color transformation in strawberry flesh. Additionally, we have identified novel genetic resources that can be instrumental in advancing research related to strawberry color change.

Anthocyanins, key quality components of tea, act as an important bridge between plants and the environment due to their function on protecting plants from biotic and abiotic irritants. This study aimed to assess the interactions between anthocyanins metabolism and the environment. Purple (P) and green (G) leaves with different anthocyanin contents were inoculated with tea plant anthracnose. High-throughput metabolomics and 16S microbial diversity sequencing methods were used to screen the anthocyanin fractions of tea plant leaves responsive to anthracnose. The interconnections between metabolites and the resistance of phyllosphere microorganisms to fungal pathogens were then analyzed. The results showed that leaves with high anthocyanin content (0.14% of diseased area ratio) were less impacted by anthracnose infestation than leaves with low anthocyanin (3.12%). The cyanidin content decreased after infection in purple leaves (PR) and increased in green leaves (GR). The relative abundance of Cyanobacteria was suppressed by the significant enrichment of Proteobacteria after anthracnose infection in green leaves. However, there were no significant differences between these two groups of microorganisms in purple leaves. Collinear network analysis revealed a strong correlation between Cyanobacteria and Dihydrosorbinol and between Proteobacteria and cyanidin metabolites. Among them, OTU456 (Bosea) was identified as the key taxonomic group of bacterial communities in the green-infected leaf network. In summary, the anthracnose resistance benefits from cyanidin mediated by proteobacteria in tea plants. These results deepen our understanding of the regulation of secondary metabolism in tea plants and the formation of plant resistance.

Brown adipose tissue (BAT) plays an important role in energy metabolism because it uses fatty acids for thermogenesis during cold exposure. Preclinical studies found that boysenberry anthocyanins (BoyACs) activate BAT. Therefore, the aim of this preliminary study was to evaluate how BoyAC intake affects BAT in humans. We performed an open-label single-arm nonrandomized study in healthy volunteers. Before and after 4 weeks of daily consumption of 100 ml boysenberry juice (BoyJ) containing 61 mg of BoyACs, participants were assessed at 24 °C and then after 1 h of mild cold exposure (18 °C). An infrared thermography camera was used to measure skin surface temperatures in the supraclavicular BAT region (Tscv) and the non-BAT region of the upper chest (Tch). Energy metabolism was measured by indirect calorimetry. For each endpoint, we calculated Δ as the difference between values before and after cold exposure and compared the values before and after BoyJ intake. 10 volunteers participated (age: 36.1 ± 4.1, body mass index (BMI): 20.9 ± 0.6). After BoyJ intake, ΔTscv-ch was significantly higher (p = 0.029), but Δ energy expenditure, Δ fat oxidation, and Δ carbohydrate oxidation were not significantly different. We found a significant positive correlation between BMI and Δfat oxidation with BoyJ intake. The results indicate that 4 weeks of BoyJ intake activates cold-induced thermogenesis in the scv-BAT but does not have a significant effect on energy metabolism. BoyJ intake may increase fat oxidation during cold exposure in individuals with higher BMI.Trial registry number: UMIN000043476, 05/03/2021., (© 2024. The Author(s).)

Fruit ripening is a highly coordinated process involving molecular and biochemical changes that collectively determine fruit quality. The underlying metabolic programs and their transitions leading to fruit ripening remain largely under-characterized in blueberry (Vaccinium sp.), which exhibits atypical climacteric behavior. In this study, we focused on sugar, acid and anthocyanin metabolism in two rabbiteye blueberry cultivars, Premier and Powderblue, during fruit development and ripening. Concentrations of the three major sugars, sucrose (Suc), glucose (Glc), and fructose (Fru) increased steadily during fruit development leading up to ripening, and increased dramatically by around 2-fold in 'Premier' and 2- to 3-fold in 'Powderblue' during the final stage of fruit ripening. Starch concentration was very low throughout fruit development in both cultivars indicating that it does not serve the role of a major transitory carbon (C) storage form in blueberry fruit. Together, these patterns indicate continued import of C, likely in the form of Suc, throughout blueberry fruit development. Concentrations of the predominant acids, malate and quinate, decreased during ripening, and may contribute to increased shikimate biosynthesis which, in-turn, allows for downstream phenylpropanoid metabolism leading to anthocyanin synthesis. Consistently, anthocyanin concentrations were highest in fully ripened blue fruit. Weighted gene co-expression network analysis (WGCNA) was performed using a 'Powderblue' fruit ripening transcriptome and targeted fruit metabolite concentration data. A 'dark turquoise' module positively correlated with sugars and anthocyanins, and negatively correlated with acids (malate, quinate), was identified. Gene Ontology (GO) enrichment analysis of this module identified transcripts related to sugar, acid, and phenylpropanoid metabolism pathways. Among these, increased transcript abundance of a VACUOLAR INVERTASE during ripening was consistent with sugar storage in the vacuole. In general, transcript abundance of the glycolysis pathway genes was upregulated during ripening. The transcript abundance of PHOSPHOENOLPYRUVATE (PEP) CARBOXYKINASE increased during fruit ripening and was negatively correlated with malate concentration, suggesting increased malate conversion to PEP, which supports anthocyanin production during fruit ripening. This was further supported by the co-upregulation of several anthocyanin biosynthesis-related genes. Together, this study provides insights into important metabolic programs, and their underlying gene expression patterns during fruit development and ripening in blueberry., (© 2024. The Author(s).)

The species-rich cosmopolitan genus Rhododendron offers a good system for exploring the genomic mechanisms underlying adaptation to diverse habitats. Here, we report high-quality chromosomal-level genome assemblies of nine species, representing all five subgenera, different altitudinal distributions, and all flower color types of this genus. Further comprehensive genomic analyses indicate diverse adaptive strategies employed by Rhododendron, particularly adaptation to alpine and subalpine habitats by expansion/contraction of gene families involved in pathogen defense and oxidative phosphorylation, genomic convergent evolution, and gene copy-number variation. The convergent adaptation to high altitudes is further shown by population genomic analysis of R. nivale from the Himalaya-Hengduan Mountains. Moreover, we identify the genes involved in the biosynthesis of anthocyanins and carotenoids, which play a crucial role in shaping flower color diversity and environmental adaptation. Our study is significant for comprehending plant adaptive evolution and the uneven distribution of species diversity across different geographical regions., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Blueberries ( Vaccinium spp.) are extremely sensitive to drought stress. Flavonoids are crucial secondary metabolites that possess the ability to withstand drought stress. Therefore, improving the drought resistance of blueberries by increasing the flavonoid content is crucial for the development of the blueberry industry. To explore the underlying molecular mechanism of blueberry in adaptation to drought stress, we performed an integrated analysis of the metabolome and transcriptome of blueberry leaves under drought stress. We found that the most enriched drought-responsive genes are mainly involved in flavonoid biosynthesis and plant hormone signal transduction pathways based on transcriptome data and the main drought-responsive metabolites come from the flavonoid class based on metabolome data. The UDP-glucose flavonoid 3-O-glucosyl transferase ( UFGT ), flavonol synthase ( FLS ), and anthocyanidin reductase ( ANR-2 ) genes may be the key genes for the accumulation of anthocyanins, flavonols, and flavans in response to drought stress in blueberry leaves, respectively. Delphinidin 3-glucoside and delphinidin-3-O-glucoside chloride may be the most important drought-responsive flavonoid metabolites. VcMYB1 , VcMYBPA1 , MYBPA1.2 , and MYBPA2.1 might be responsible for drought-induced flavonoid biosynthesis and VcMYB14, MYB14, MYB102 , and MYB108 may be responsible for blueberry leaf drought tolerance. ABA responsive elements binding factor ( ABF ) genes, MYB genes, bHLH genes, and flavonoid biosynthetic genes might form a regulatory network to regulate drought-induced accumulation of flavonoid metabolites in blueberry leaves. Our study provides a useful reference for breeding drought-resistant blueberry varieties.

Background: Spine grape (Vitis davidii) is a promising source of high-quality anthocyanins, with vast potential for application in food, pharmaceutical, and cosmetic industries. However, their availability is limited by resource constraints. Plant cell culture has emerged as a valuable approach for anthocyanin production and serves as an ideal model to investigate the regulation of anthocyanin biosynthesis. Elicitors are employed to achieve targeted enhancement of anthocyanin biosynthesis. The present study investigated the impact of 5-aminolevulinic acid (ALA) as an elicitor on the accumulation of anthocyanins and flavonoids during spine grape callus growth. Specifically, we examined the effects of ALA on anthocyanin and its component accumulation in callus, and biosynthetic anthocyanin gene expression., Results: ALA at 25 µg/L increased the biomass of spine grape callus. ALA induction enhanced the levels of flavonoids, anthocyanins and proanthocyanidins in callus, with maximum values reaching 911.11 mg/100 g DW, 604.60 mg/100 g DW, and 5357.00 mg/100 g DW, respectively, after callus culture for 45 days. Notably, those levels were 1.47-, 1.93- and 1.83-fold higher than controls. ALA induction modulated the flavonoid profile, and among 97 differential flavonoid metabolites differing from controls, 77 were upregulated and 20 were downregulated. Six kinds of anthocyanins, namely cyanidin (8), delphinidin (6), peonidin (5), malvidin (4), petunidin (3) and pelargonidin (3), were detected in callus, with peonidin most abundant. Compared with controls, anthocyanin components were increased in ALA-treated callus. The key genes PAL1, PAL2, PAL4, CHI, CHS3, F3'H, F3H, FLS, DFR, UFGT, MYBA1, LDOX, OMT3, GT1 and ACT involved in anthocyanin biosynthesis were upregulated following ALA treatment, resulting in anthocyanin accumulation., Conclusion: This study revealed a novel mode of ALA-mediated promotion of plant anthocyanin biosynthesis and accumulation at the cellular level, and a strategy for enhancing anthocyanin content in spine grape callus. The findings advance commercial-scale production of anthocyanins via spine grape callus culture. we also explored the accumulation patterns of flavonoids and anthocyanins under ALA treatment. Augmentation of anthocyanins coincided with elevated expression levels of most genes involved in anthocyanin biosynthesis within spine grape callus following ALA treatment., (© 2024. The Author(s).)

Horticulture in controlled environments has been increasingly used to tackle limitations on crop production. As a crucial environmental factor, light regulate plant growth and metabolism. In the present study, basil plants were subjected to different light durations and intensities considering constant daily light integral (DLI). The lighting environment included 200, 300, and 400 µmol m - 2 s - 1 intensities for 18, 12, and 9 h, respectively. DLI amounted to 12.96 mol m - 2 d - 1 among all light treatments (LI200 for 18 h, LI300 for 12 h, and LI400 for 9 h). Half of the plants under each light treatment were exposed to 30 µmol m - 2 s - 1 of far-red light. The results indicated the general negative impact of LI400/9 on the growth of basils. Exposure to far-red light hurt the growth of the shoot, while it enhanced stem and petiole elongation. This effect was due to higher gibberellin accumulation, which resulted in shade avoidance responses. Exposure to far-red light also reduced anthocyanin and flavonoid contents, as two important nutritional components. Soluble carbohydrates increased, while storage carbohydrates decreased by increasing lighting duration/decreasing light intensity or by far-red light inclusion. The lowest antioxidant activity was detected in LI400/9. In the LI200/18, the highest level of auxin and the lowest level of cytokinin were detected, while the LI300/12 exhibited the highest level of gibberellin hormone. Low light intensity and long photoperiod enhanced plant biomass and phytochemical production and are recommended for basil production in controlled environments., (© 2024. The Author(s).)

The color of coffee fruits is influenced by several factors, including cultivar, ripening stage, and metabolite composition. However, the metabolic accumulation of pigments and the molecular mechanisms underlying peel coloration during the ripening process of Coffea arabica L. remain relatively understudied. In this study, UPLC-MS/MS-based metabolomics and RNA sequencing (RNA-seq)-based transcriptomics were integrated to investigate the accumulation of anthocyanins and carotenoids in the peel of Coffea arabica at different ripening stages: green peel (GP), green-yellow peel (GYRP), red peel (RP), and red-purple peel (RPP). This integration aimed at elucidating the molecular mechanisms associated with these changes. A total of ten anthocyanins, six carotenoids, and thirty-five xanthophylls were identified throughout the ripening process. The results demonstrated a gradual decrease in the total carotenoid content in the peel with fruit maturation, while anthocyanin content increased significantly. Notably, the accumulation of specific anthocyanins was closely associated with the transition of peel colors from green to red. Integrated metabolomics and transcriptomics analyses identified the GYRP stage as critical for this color transition. A weighted gene co-expression network analysis (WGCNA) revealed that enzyme-coding genes such as 3AT, BZ1, and lcyE, along with transcription factors including MYB, NAC, and bHLH, which interact with PHD and SET TR, may regulate the biosynthesis of anthocyanins and carotenoids, thereby influencing peel pigmentation. These findings provide valuable insights into the molecular mechanisms underlying the accumulation of anthocyanins and carotenoids in Coffea arabica peel during fruit maturation.

Background: Understanding the molecular basis of sport mutations in fruit trees has the potential to accelerate generation of improved cultivars., Results: For this, we analyzed the genome of the apple tree that developed the RubyMac phenotype through a sport mutation that led to the characteristic fruit coloring of this variety. Overall, we found 46 somatic mutations that distinguished the mutant and wild-type branches of the tree. In addition, we found 54 somatic gene conversions (i.e., loss-of-heterozygosity mutations) that also distinguished the two parts of the tree. Approximately 20% of the mutations were specific to individual cell lineages, suggesting that they originated from the corresponding meristematic layers. Interestingly, the de novo mutations were enriched for GC =  > AT transitions while the gene conversions showed the opposite bias for AT =  > GC transitions, suggesting that GC-biased gene conversions have the potential to counteract the AT-bias of de novo mutations. By comparing the gene expression patterns in fruit skins from mutant and wild-type branches, we found 56 differentially expressed genes including 18 involved in anthocyanin biosynthesis. While none of the differently expressed genes harbored a somatic mutation, we found that some of them in regions of the genome that were recently associated with natural variation in fruit coloration., Conclusion: Our analysis revealed insights in the characteristics of somatic change, which not only included de novo mutations but also gene conversions. Some of these somatic changes displayed strong candidate mutations for the change in fruit coloration in RubyMac., (© 2024. The Author(s).)

Background: Polymorphisms are common in nature, but they are rarely shared among closely related species. Polymorphisms could originate through convergence, ancestral polymorphism, or introgression. Although shared neutral genomic variation across species is commonplace, few examples of shared functional traits exist. The blue-orange petal color polymorphisms in two closely related species, Lysimachia monelli and L. arvensis were investigated with UV-vis reflectance spectra, flavonoid biochemistry, and transcriptome comparisons followed by climate niche analysis., Results: Similar color morphs between species have nearly identical reflectance spectra, flavonoid biochemistry, and ABP gene expression patterns. Transcriptome comparisons reveal two orange-specific genes directly involved in both blue-orange color polymorphisms: DFR-2 specificity redirects flux from the malvidin to the pelargonidin while BZ1-2 stabilizes the pelargonidin with glucose, producing the orange pelargonidin 3-glucoside. Moreover, a reduction of F3'5'H expression in orange petals also favors pelargonidin production. The climate niches for each color morph are the same between the two species for three temperature characteristics but differ for four precipitation variables., Conclusions: The similarities in reflectance spectra, biochemistry, and ABP genes suggest that a single shift from blue-to-orange shared by both lineages is the most plausible explanation. Our evidence suggests that this persistent flower color polymorphism may represent an ancestrally polymorphic trait that has transcended speciation, yet future analyses are necessary to confidently reject the alternative hypotheses., (© 2024. The Author(s).)

Background: Oil palm (Elaeis guineensis Jacq.) is a very important tropical woody oil plant with high commercial and ornamental value. The exocarp of the oil palm fruit is rich in anthocyanosides and proanthocyanidins, which not only give it a bright colour, but also mark the maturity of the fruit. The study of the dynamic change pattern of anthocyanoside content and important anthocyanoside metabolism-related regulatory genes during oil palm ripening is conducive to the improvement of the ornamental value of oil palm and the determination of the optimal harvesting period of the fruits., Methods: We analyzed the virescens oil palm (AS) and nigrescens oil palm (AT) at 95 days (AS1, AT1), 125 days (AS2, AT2) and 185 days (AS3, AT3) after pollination were used as experimental materials for determining the changes in the total amount of anthocyanins as well as their metabolomics and transcriptomics studies by using the LC-MS/MS technique and RNA-Seq technique., Result: The results showed that the total anthocyanin content decreased significantly from AS1 (119 µg/g) to AS3 (23 µg/g), and from AT1 (1302 µg/g) to AT3 (170 µg/g), indicating a clear decreasing trend during fruit development. Among them, the higher flavonoids in AS and AT included anthocyanins such as peonidin-3-O-rutinoside (H35), pelargonidin-3-O-rutinoside (H21), and cyanidin-3-O-glucoside (H7), as well as condensed tannins such as procyanidin B2 (H47), procyanidin C1 (H49), and procyanidin B3 (H48). Notably, nine genes involved in the anthocyanin biosynthetic pathway exhibited up-regulated expression during the pre-development stage of oil palm fruits, particularly during the AS1 and AT1 periods. These genes include: Chalcone synthase (CHS; LOC105036364); Flavanone 3-hydroxylase (F3H; LOC105054663); Dihydroflavonol 4-reductase (DFR; LOC105040724, LOC105048473); Anthocyanidin synthase (ANS; LOC105035842), UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT; LOC105039612); Flavonoid 3',5'-hydroxylase (F3'5'H; LOC105036086, LOC105044124, LOC105045493). In contrast, five genes demonstrated up-regulated expression as the fruits developed, specifically during the AS3 and AT3 periods. These genes include: Chalcone synthase (CHS; LOC105036921, LOC105035716); Chalcone isomerase (CHI; LOC105045978); UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT; LOC105046326); Flavonoid 3'-hydroxylase (F3'H; LOC105036086)., Conclusion: Most of differentially expressed genes exhibited up-regulation during the early stages of fruit development, which may contribute to the elevated anthocyanin content observed in oil palm fruits of both types during the pre-developmental period. Furthermore, the expression levels of most genes were found to be higher in the AT fruit type compared to the AS fruit type, suggesting that the differential expression of these genes may be a key factor underlying the differences in anthocyanoside production in the exocarp of oil palm fruits from these two fruit types. The findings of this study provide a theoretical foundation for the identification and characterization of genes involved in anthocyanin synthesis in oil palm fruits, as well as the development of novel variations using molecular biology approaches., (© 2024. The Author(s).)

Microalgae like Chlamydomonas are beneficial organisms employed as biological stimulants to improve plants' growth, fruit quality, and stress tolerance. In the current study, the effects of Chlamydomonas sp. foliar spraying (0, 20, and 40 ml L -1 ) were assayed on Camarosa strawberry plants under salinity stress (0, 40, and 80 mM NaCl). The results showed that the foliar application of Chlamydomonas extract influenced strawberry's morphological, physiological, and biochemical characteristics under salinity stress. Foliar treatment of Chlamydomonas extract with and without salinity stress increased the leaf number and leaf area, the leaf relative water content, and photosynthetic pigments content. Moreover, the foliar application of Chlamydomonas extract decreased lipid peroxidation and hydrogen peroxide content and, on the other hand, enhanced the antioxidant enzymes activity (superoxide dismutase, guaiacol peroxidase, and peroxidase), phenolics, flavonoids, and anthocyanins content under salinity stress. For instance, the highest total antioxidant capacity was found in the plants foliar treated with 40 ml L -1  of Chlamydomonas algae extract under 80 mM salinity stress, which increased by 102.4% compared to the controls, as well as the highest total phenolic compounds and anthocyanin's content were 30.22, and 7.2% more than the control plants, respectively. Overall, the foliar application of Chlamydomonas algae extracts, especially at a concentration of 20 ml L -1  enhanced the strawberry's growth, yield, and physiological traits under saline conditions. The results with more detailed evaluations will be advisable for the pioneer farmers and extension section., (© 2024. The Author(s).)

Background: While water availability is important for quality at harvest, it also continues to influence the quality of pomegranates during storage. Reducing the amount of irrigation, in addition to water saving has different effects on bioactive compounds of pomegranate during storage time. This study was conducted to determine the influence of irrigation level on fruit quality changes during storage period of two commercial Iranian pomegranate cultivars ('Shishecap' and 'Malas-Yazdi'). Sustained deficit irrigation (SDI) was applied to plants that received 75% (moderate stress) or 50% (severe stress) of their normal water requirement. A control group received 100% of their water requirement., Results: At harvest time and during storage period, fruit weight loss and some biochemical traits such as fruit total soluble solids (TSS), titratable acidity (TA), pH, total phenolic compounds (TPC), total anthocyanins content (TAC), antioxidant activity and vitamin C were measured in pomegranate fruits. Also, the quantity of the produced product was also measured at the time of harvesting. Results indicated that control fruits exhibited more weight loss than those produced under water deficit during the storage period in both years. According to results, fruit TSS, TAC, and antioxidant activity significantly increased during storage period but fruit TA and vitamin C significantly decreased throughout storage period. Also, reduction in irrigation level resulted in a decline in the yield., Conclusions: This study revealed a crucial link between irrigation level and the quality of pomegranate fruits, despite a reduction in the yield. This included affecting weight loss and the content of bioactive compounds, both at harvest and during storage., (© 2024. The Author(s).)

Anthocyanins are bioactive compounds responsible for various physiological processes in plants and provide characteristic colors to fruits and flowers. Their biosynthetic pathway is well understood; however, the enzymatic degradation mechanism is less explored. Anthocyanase (β-glucosidase (BGL)), peroxidase (POD), and polyphenol oxidase (PPO) are enzymes involved in degrading anthocyanins in plants such as petunias, eggplants, and Sicilian oranges. The aim of this work was to investigate the physicochemical interactions between these enzymes and the identified anthocyanins (via UPLC-MS/MS) in cranberry ( Vaccinium macrocarpon ) through molecular docking to identify the residues likely involved in anthocyanin degradation. Three-dimensional models were constructed using the AlphaFold2 server based on consensus sequences specific to each enzyme. The models with the highest confidence scores (pLDDT) were selected, with BGL, POD, and PPO achieving scores of 87.6, 94.8, and 84.1, respectively. These models were then refined using molecular dynamics for 100 ns. Additionally, UPLC-MS/MS analysis identified various flavonoids in cranberries, including cyanidin, delphinidin, procyanidin B2 and B4, petunidin, pelargonidin, peonidin, and malvidin, providing important experimental data to support the study. Molecular docking simulations revealed the most stable interactions between anthocyanase and the anthocyanins cyanidin 3-arabinoside and cyanidin 3-glucoside, with a favorable ΔG of interaction between -9.3 and -9.2 kcal/mol. This study contributes to proposing a degradation mechanism and seeking inhibitors to prevent fruit discoloration.

Basic leucine zipper (bZIP) transcription factors (TFs) play a crucial role in anthocyanin accumulation in plants. In addition to bZIP TFs, abscisic acid (ABA) increases anthocyanin biosynthesis. Therefore, this study aimed to investigate whether bZIP TFs are involved in ABA-induced anthocyanin accumulation in sweet cherry and elucidate the underlying molecular mechanisms. Specifically, the BLAST method was used to identify bZIP genes in sweet cherry. Additionally, we examined the expression of ABA- and anthocyanin-related genes in sweet cherry following the overexpression or knockdown of a bZIP candidate gene. In total, we identified 54 bZIP -encoding genes in the sweet cherry genome. Basic leucine zipper 6 ( bZIP6 ) showed significantly increased expression, along with increased anthocyanin accumulation in sweet cherry. Additionally, yeast one-hybrid and dual-luciferase assays indicated that PavbZIP6 enhanced the expression of anthocyanin biosynthetic genes ( PavDFR , PavANS , and PavUFGT ), thereby increasing anthocyanin accumulation. Moreover, PavbZIP6 interacted directly with the PavBBX6 promoter, thereby regulating PavNCED1 to promote abscisic acid (ABA) synthesis and enhance anthocyanin accumulation in sweet cherry fruit. Conclusively, this study reveals a novel mechanism by which PavbZIP6 mediates anthocyanin biosynthesis in response to ABA and contributes to our understanding of the mechanism of bZIP genes in the regulation of anthocyanin biosynthesis in sweet cherry.

Dihydroflavonol 4-reductase (DFR) significantly influences the modification of flower color. To explore the role of DFR in the synthesis of strawberry anthocyanins, in this study, we downloaded the CDS sequences of the DFR gene family from the Arabidopsis genome database TAIR; the DFR family of forest strawberry was compared; then, a functional domain screen was performed using NCBI; the selected strawberry DFR genes were analyzed; and the expression characteristics of the family members were studied by qRT-PCR. The results showed that there are 57 members of the DFR gene family in strawberry, which are mainly expressed in the cytoplasm and chloroplast; most of them are hydrophilic proteins; and the secondary structure of the protein is mainly composed of α-helices and random coils. The analysis revealed that FvDFR genes mostly contain light, hormone, abiotic stress, and meristem response elements. From the results of the qRT-PCR analysis, the relative expression of each member of the FvDFR gene was significantly different, which was expressed throughout the process of fruit coloring. Most genes had the highest expression levels in the full coloring stage (S4). The expression of FvDFR30 , FvDFR54 , and FvDFR56 during the S4 period was 8, 2.4, and 2.4 times higher than during the S1 period, indicating that the DFR gene plays a key role in regulating the fruit coloration of strawberry. In the strawberry genome, 57 members of the strawberry DFR gene family were identified. The higher the DFR gene expression, the higher the anthocyanin content, and the DFR gene may be the key gene in anthocyanin synthesis. Collectively, the DFR gene is closely related to fruit coloring, which lays a foundation for further exploring the function of the DFR gene family.

Long non-coding RNAs (lncRNAs), a class of important regulatory factors for many biological processes in plants, have received much attention in recent years. To explore the molecular roles of lncRNAs in sweet cherry fruit ripening, we conducted widely targeted metabolome, transcriptome and lncRNA analyses of sweet cherry fruit at three ripening stages (yellow stage, pink stage, and dark red stage). The results show that the ripening of sweet cherry fruit involves substantial metabolic changes, and the rapid accumulation of anthocyanins (cyanidin 3-rutinoside, cyanidin 3-O-galactoside, and cyanidin 3-O-glucoside) is the main cause of fruit coloration. These ripening-related alterations in the metabolic profile are driven by specific enzyme genes related to the synthesis and decomposition of abscisic acid (ABA), cell wall disintegration, and anthocyanin biosynthesis, as well as transcription factor genes, such as MYBs , bHLHs , and WD40s . LncRNAs can target these ripening-related genes to form regulatory modules, incorporated into the sweet cherry fruit ripening regulatory network. Our study reveals that the lncRNA-mRNA module is an important component of the sweet cherry fruit ripening regulatory network. During sweet cherry fruit ripening, the differential expression of lncRNAs will meditate the spatio-temporal specific expression of ripening-related target genes (encoding enzymes and transcription factors related to ABA metabolism, cell wall metabolism and anthocyanin metabolism), thus driving fruit ripening.

Background: The Flavonoid 3'-hydroxylase gene(F3'H) is an important structural gene in the anthocyanin synthesis pathway of plants, which has been proven to be involved in the color formation of organs such as leaves, flowers, and fruits in many plants. However, the mechanism and function in barley are still unclear., Results: In order to explore the molecular mechanism of the grain color formation of purple qingke, we used the cultivated qingke variety Nierumzha (purple grain) and the selected qingke variety Kunlun 10 (white grain) to conduct transcriptomic sequencing at the early milk, late milk and soft dough stage. Weighted Gene Co-expression Network Analysis (WGCNA) was used to construct weighted gene co-expression network related to grain color formation, and three key modules (brown, yellow, and turquoise modules) related to purple grain of qingke were selected. F3'H (HORVU1Hr1G094880) was selected from the hub gene of the module for the yeast library, yeast two-hybrid (Y2H), subcellular localization and other studies. It was found that in purple qingke, HvnF3'H mainly distributed in the cytoplasm and cell membrane and interacted with several stress proteins such as methyltransferase protein and zinc finger protein., Conclusions: The results of this study provide reference for the regulation mechanism of anthocyanin-related genes in purple grain qingke., (© 2024. The Author(s).)

Indoor farming systems enable plant production in precisely controlled environments. However, implementing stable growth conditions and the absence of stress stimulants can weaken plants' defense responses and limit the accumulation of bioactive, health-beneficial phytochemicals. A potential solution is the controlled application of stressors, such as supplemental ultraviolet (UV) light. To this end, we analyzed the efficiency of short-term pre-harvest supplementation of the red-green-blue (RGB, LED) spectrum with ultraviolet B (UV-B) or C (UV-C) light to boost phytochemical synthesis. Additionally, given the biological harm of UV radiation due to high-energy photons, we monitored plants' photosynthetic activity during treatment and their morphology as well as sensory attributes after the treatment. Our analyses showed that UV-B radiation did not negatively impact photosynthetic activity while significantly increasing the overall antioxidant potential of lettuce through enhanced levels of secondary metabolites (total phenolics, flavonoids, anthocyanins), carotenoids, and ascorbic acid. On the contrary, UV-C radiation-induced anthocyanin accumulation in the green leaf cultivar significantly harmed the photosynthetic apparatus and limited plant growth. Taken together, we showed that short-term UV-B light supplementation is an efficient method for lettuce biofortification with healthy phytochemicals, while UV-C treatment is not recommended due to the negative impact on the quality (morphology, sensory properties) of the obtained leafy products. These results are crucial for understanding the potential of UV light supplementation for producing functional plants.