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Seagrasses are a paraphyletic group of marine angiosperms and retain certain adaptations from the ancestors of all embryophytes in the transition to terrestrial environments. Among these adaptations is the production of flavonoids, versatile phenylpropanoid secondary metabolites that participate in a variety of stress responses. Certain features, such as catalytic promiscuity and metabolon interactions, allow flavonoid metabolism to expand to produce novel compounds and respond to a variety of stimuli. As marine environments expose seagrasses to a unique set of stresses, these plants display interesting flavonoid profiles, the functions of which are often not completely clear. Flavonoids will likely prove to be effective and versatile agents in combating the new host of stress conditions introduced to marine environments by anthropogenic climate change, which affects marine environments differently from terrestrial ones. These new stresses include increased sulfate levels, changes in salt concentration, changes in herbivore distributions, and ocean acidification, which all involve flavonoids as stress response mechanisms, though the role of flavonoids in combatting these climate change stresses is seldom discussed directly in the literature. Flavonoids can also be used to assess the health of seagrass meadows through an interplay between flavonoid and simple phenolic levels, which may prove to be useful in monitoring the response of seagrasses to climate change. Studies focusing on the genetics of flavonoid metabolism are limited for this group, but the large chalcone synthase gene families in some species may provide an interesting topic of research. Anthocyanins are typically studied separately from other flavonoids. The phenomenon of reddening in certain seagrass species typically focuses on the importance of anthocyanins as a UV-screening mechanism, while the role of anthocyanins in cold stress is discussed less often. Both of these stress response functions would be useful for adaptation to climate change-induced deviations in tidal patterns and emersion. However, ocean warming will likely lead to a decrease in anthocyanin content, which may impact the performance of intertidal seagrasses. This review highlights the importance of flavonoids in angiosperm stress response and adaptation, examines research on flavonoids in seagrasses, and hypothesizes on the importance of flavonoids in these organisms under climate change. [ABSTRACT FROM AUTHOR]

Introduction: Silybum marianum is a medicinal plant that produces silymarin, which has been demonstrated to possess antiviral, anti-neurodegenerative, and anticancer activities. Silybin (A+B) are two major hepatoprotective flavonolignans produced predominantly in S. marianum fruits. Several attempts have been made to increase the synthesis of silymarin, or its primary components, silybin (A+B). Zinc oxide nanoparticles (ZnO-NPs) are considered a highly efficient Zn source widely used to promote crop development and productivity. Methods: In this study, we aimed to investigate the effects of the foliar application of ZnO-NPs on the growth, yield, photosynthetic pigment content, silybin (A+B) content, and the expression of the chalcone synthase (CHS) gene in S. marianum plants. Different concentrations of ZnO-NPs were administered as foliar sprays to S. marianum plants growing in greenhouse conditions. Furthermore, we evaluated the anti-osteoporotic efficacy of the corresponding fruit extract against dexamethasone (Dex)-induced osteoporosis. Results and discussion: Foliar treatment at all ZnO-NP concentrations increased the amounts of bioactive components of silybin (A+B), which enhanced the growth and yield of S. marianum plants while increasing the levels of N, P, K, and Zn in their leaves, roots, and fruits; the levels of photosynthetic pigments in their leaves; and silybin (A+B) content in their fruits, thereby increasing the medicinal value of S. marianum. The highest gains were observed in plants sprayed with the highest ZnO-NP concentration (20.0 mg/L). In addition, gene expression studies revealed that ZnO-NPs stimulated silybin (A+B) production by activating CHS genes. The administration of S. marianum extracts to Dex-administered rats increased osteoblast and bone formation while inhibiting osteoclast and bone resorption, thereby protecting the animals against Dex-induced osteoporosis. [ABSTRACT FROM AUTHOR]

Djulis (Chenopodium formosanum Koidz.), a member of the Amaranthaceae family plant, is noted for its vibrant appearance and significant ornamental value. However, the mechanisms underlying color variation in its spikes remain unexplored. This research initially detected the anthocyanin content at different developmental stages of the spike and subsequently utilized an integrative approach, combining targeted metabolomics, transcriptomics, and untargeted metabolomics analyses, to elucidate the mechanisms of anthocyanin biosynthesis in the spikes of djulis. The results of the combined multi-omics analysis showed that the metabolites associated with anthocyanin synthesis were mainly enriched in the flavonoid biosynthesis pathway (ko00941) and the anthocyanin biosynthesis pathway (ko00942). With the maturation of djulis spikes, a total of 28 differentially expressed genes and 17 differentially expressed metabolites were screened during the transition of spike color from green (G) to red (R) or orange (O). Twenty differentially expressed genes were selected for qRT-PCR validation, and the results are consistent with transcriptome sequencing. The upregulation of seven genes, including chalcone synthase (CfCHS3_1, CfCHS3_2, CfCHS3_3), flavanone 3-hydroxylase (CfF3H_3), flavonoid 3′5′-hydroxylase (CfCYP75A6_1), dihydroflavonol reductase (CfDFRA), and glucosyltransferase (Cf3GGT), promotes the formation and accumulation of delphinidin 3-sambubioside and peonidin 3-galactoside. The research results also showed that anthocyanins and betalains can coexist in the spike of djulis, and the reason for the change in spike color during development may be the result of the combined action of the two pigments. A possible regulatory pathway for anthocyanin biosynthesis during the spike maturation was constructed based on the analysis results. The results provide a reference and theoretical basis for further studying the molecular mechanism of anthocyanin regulation of color changes in Amaranthaceae plants. [ABSTRACT FROM AUTHOR]

Chalcone synthase (CHS), the first key structural enzyme in the flavonoid biosynthesis pathway, plays a crucial role in regulating plant responses to abiotic stresses and hormone signaling. However, its molecular functions remain largely unknown in Phyllostachys edulis, which is one of the most economically and ecologically important bamboo species and the most widely distributed one in China. This study identified 17 CHS genes in Phyllostachys edulis and classified them into seven subgroups, showing a closer evolutionary relationship to CHS genes from rice. Further analysis of PeCHS genes across nine scaffolds revealed that most expansion occurred through tandem duplications. Collinearity analysis indicated strong evolutionary conservation among CHS genes. Motif and gene structure analyses confirmed high structural similarity, suggesting shared functional characteristics. Additionally, cis-acting element analysis demonstrated that PeCHS genes are involved in hormonal regulation and abiotic stress responses. RNA-Seq expression profiles in different bamboo shoot tissues and heights, under various hormone treatments (gibberellin (GA), naphthaleneacetic acid (NAA), abscisic acid (ABA), and salicylic acid (SA)), as well as salinity and drought stress, revealed diverse response patterns among PeCHS genes, with significant differential expression, particularly under hormone treatments. Notably, PeCHS14 consistently maintained high expression levels, suggesting its key role in stress response mechanisms. qRT-PCR analysis further validated the expression differences in five PeCHS genes under GA and ABA treatments. Subcellular localization analysis demonstrated that PeCHS14 and PeCHS15 proteins are localized in the nucleus. This study provides a foundation for investigating the potential functions of PeCHS genes and identifies candidate genes for future research on the responses of Phyllostachys edulis to abiotic stresses and hormone signaling. [ABSTRACT FROM AUTHOR]

A network model was constructed to illustrate the potential regulatory mechanism through which LED influences postharvest ripening and senescence of apricot fruits based on the comprehensive analysis of transcriptome, metabolome and ATAC-Seq. [Display omitted] • LED treatment effectively maintained the quality of apricot fruit during postharvest storage. • LED treatment activated pathways involved in ascorbate and aldarate metabolism, flavonoid biosynthesis. • LED treatment activated pathways involved in ascorbate and aldarate metabolism, flavonoid biosynthesis.LED treatment can promote phenylpropanoid and anthocyanin biosynthesis. • LED treatment influenced the expression of genes associated with plant hormone signal transduction, fruit texture and color transformation, and plant antioxidant activity. Apricot (Prunus armeniaca L.) fruits are highly perishable and prone to quality deterioration during storage and transportation. To investigate the effects of LED white light treatment on postharvest ripening of fruits using metabolomics, transcriptomics, and ATAC-Seq analysis. Fruits were exposed to 5 μmol m−2 s−1 LED white light for 12 h followed by 12 h of darkness at 20 °C daily for 12 days. The effects of the treatments on the physiological and nutritional quality of the fruits were evaluated. These data were combined with transcriptomic, metabolomic, and ATAC-Seq data from fruits taken on 8 d of treatment to provide insight into the potential mechanism by which LED treatment delays ripening. LED treatment activated pathways involved in ascorbate and aldarate metabolism and flavonoid and phenylpropanoid biosynthesis. Specifically, LED treatment increased the expression of UDP-sugar pyrophosphorylase (USP) , L-ascorbate peroxidase (AO) , dihydroflavonol 4-reductase (DFR) , chalcone synthase (CHS) , and caffeoyl-CoA O-methyltransferase (CCOAOMT1) , leading to the accumulation of caffeoyl quinic acid, epigallocatechin, and dihydroquercetin and the activation of anthocyanin biosynthesis. LED treatment also affected the expression of genes associated with plant hormone signal transduction, fruit texture and color transformation, and antioxidant activity. The notable genes affected by LED treatment included 1-aminocyclopropane-1-carboxylate synthase (ACS) , 1-aminocyclopropane-1-carboxylate oxidase (ACO) , hexokinase (HK) , lipoxygenase (LOX) , malate dehydrogenase (MDH) , endoglucanase (CEL) , various transcription factors (TCP, MYB, EFR) , and peroxidase (POD). ATAC-Seq analysis further revealed that LED treatment primarily regulated phenylpropanoid biosynthesis. The results obtained in this study provide insights into the effects of LED light exposure on apricot fruits ripening. LEDs offer a promising approach for extending the shelf life of other fruits and vegetables. [ABSTRACT FROM AUTHOR]

White clover (Trifolium repens) is vulnerable to drought stress. In response to abiotic stress, plants are regulated by NAC transcription factors. The NAC in white clover has not been thoroughly documented until recently. We have identified one white clover NAC transcription factor called TrNAC002. TrNAC002's coding sequence is localized to specific regions on the 3P and 5O chromosomes of white clover and is part of a single-copy nuclear gene. Subcellular localization demonstrates that TrNAC002 is located in the nucleus, while the transcriptional activity assay indicates its transcriptional activity. Arabidopsis plants overexpressing TrNAC002 (OE) exhibit enlarged leaves and increased lateral root growth compared to the wild type (WT). Additionally, the expression levels of the shoot apical meristem (SAM), WUSCHEL (WUS), DNA-binding protein (DBP), and auxin-induced in root cultures3 (AIR3) genes are significantly higher in OE as compared to WT. These findings imply that TrNAC002 could promote vegetative growth by increasing the expression of these genes. Under natural drought stress, OE can survive in dry soil for a longer period of time than WT. Furthermore, OE exhibits a lower level of reactive oxygen species (ROS) level and a higher content of flavonoids than WT. This is also positively correlated with an increased flavonoid content. In white clover, the expression of TrNAC002, chalcone synthase (CHS), and chalcone isomerase (CHI) in leaves demonstrates significant upregulation after drought stress and ABA treatment, as does the flavonoid content. However, the pTRV-VIGS experiment suggests that pTRV2-TrNAC002 white clover shrinks compared to the Mock and Water controls. Additionally, pTRV2-TrNAC002 white clover displays a statistically higher malondialdehyde (MDA) content than the Mock and Water controls, and a significantly lower level of total antioxidant activities, flavonoid content, CHS and CHI relative expression than that of the Mock and Water controls. These findings indicate that TrNAC002 responds to drought and modulates flavonoid biosynthesis in white clover. This study is the first to suggest that TrNAC002 likely responds to drought via ABA and enhances plant drought resistance by synthesizing flavonoids. [ABSTRACT FROM AUTHOR]

Introduction: Heat stress caused by global warming adversely affects wheat yield through declining most nutritional quality attributes in grains, excluding grain protein content. Methods: This research investigated the biochemical, physiological, and antioxidant responses of wheat plants under heat stress, focusing on the role of plant growth-promoting bacteria (Micromonospora sp.). Two wheat genotypes were studied: one heat-sensitive and one heat-tolerant, examining their responses to heat stress with and without bacterial inoculation. Results: Under heat stress, the sensitive cultivar experienced significant reductions in photosynthesis rate, chlorophyll content, and RuBisCO activity (57-61%), while the tolerant cultivar had milder reductions (24-28%). Micromonospora sp. treatment notably improved these parameters in the sensitive cultivar (+48-78%), resulting in a substantial increase in biomass production (+43-53%), which was not seen in the tolerant cultivar. Additionally, oxidative stress markers (H2O2 and MDA) were elevated more in the sensitive cultivar (82% and 90% higher) compared to the tolerant one. Micromonospora sp. treatment effectively reduced these markers in the sensitive cultivar (-28% and -27%). Enhanced activity of antioxidant enzymes and ASC-GSH pathway enzymes was particularly evident in Micromonospora sp.-treated sensitive plants. Carbohydrate metabolism shifted, with increased soluble sugars and significant rises in sucrose content in Micromonospora sp.-treated plants under stress. Discussion: The higher soluble sugar levels facilitated amino acid synthesis, contributing to biosynthesis of secondary metabolites, including flavonoids, polyphenols, and anthocyanins. This was reflected in increased activity of phenylalanine ammonia-lyase, cinnamate (CA) 4-hydroxylase, and chalcone synthase enzymes, indicating the activation of phenylpropanoid pathways. Overall, the findings suggest that Micromonospora sp. can mitigate heat stress effects by enhancing photosynthetic efficiency, antioxidant defense, and metabolic adaptations in heat-sensitive wheat cultivars. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Summary Soybean [Glycine max (L.) Merr.] is an exceptionally rich in isoflavones, and these compounds attach to oestrogen receptors in the human body, lessening the risk of breast cancer and effectively alleviating menopausal syndrome symptoms. Uncovering the molecular mechanisms that regulate soybean isoflavone accumulation is crucial for enhancing the production of these compounds. In this study, we combined bulk segregant analysis sequencing (BSA‐seq) and a genome‐wide association study (GWAS) to discover a novel R2R3‐MYB family gene, GmMYB77, that regulates isoflavone accumulation in soybean. Using the soybean hairy root transient expression system, we verified that GmMYB77 inhibits isoflavone accumulation. Furthermore, knocking out GmMYB77 significantly increased total isoflavone (TIF) content, particularly malonylglycitin, while its overexpression resulted in a notable decrease in contents of malonylglycitin and TIF. We found that GmMYB77 can directly binds the core sequence GGT and suppresses the expression of the key isoflavone biosynthesis genes Isoflavone synthase 1 (GmIFS1), Isoflavone synthase 2 (GmIFS2), Chalcone synthase 7 (GmCHS7) and Chalcone synthase 8 (GmCHS8) by using dual‐luciferase assays, electrophoretic mobility shift assays and yeast one‐hybrid experiments. Natural variations in the promoter region of GmMYB77 affect its expression, thereby regulating the malonylglycitin and TIF contents. Hap‐P2, an elite haplotype, plays a pivotal role in soybean breeding for substantially enhanced isoflavone content. These findings enhance our understanding of the genes influencing soybean isoflavone content and provide a valuable genetic resource for molecular breeding efforts in the future. [ABSTRACT FROM AUTHOR]

During the storage and processing of lily bulbs, the phenomenon of the violet-red color change of the scales often occurs. To investigate the effect of temperature on the violet-red color change of fresh-cut lily bulb scales, Lilium davidii var. unicolor bulb scales were stored at different temperatures (4, 20, 35 °C), and the surface color, total phenolic content, total anthocyanin content, the activities of anthocyanin synthesis-related enzymes, polyphenol oxidase (PPO), and peroxidase (POD) were determined. Results indicated that higher temperatures increased the total phenolic content of lily bulb scales during storage. In the middle of the storage period, temperature promoted the activity of polyphenol oxidase (PPO) and peroxidase (POD), leading to an increase in the browning degree of lily bulb scales and a decrease in surface quality. At the same time, higher temperatures promoted the activity of anthocyanin synthesis-related enzymes, such as phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), dihydroflavonol reductase (DFR), and anthocyanin synthase (ANS), and a storage temperature of 20 °C could significantly increase the accumulation of anthocyanins in lily scales, but it was not enough to cause a violet-red color on the surface of lily bulb scales. Overall, the temperature is not the main factor affecting the violet-red color change of fresh-cut lily bulb scales. [ABSTRACT FROM AUTHOR]

Abiotic stress not only elevates the synthesis of secondary metabolites in plant sprouts but also boosts their antioxidant capacity. In this study, the mechanisms of flavonoid biosynthesis and antioxidant systems in buckwheat sprouts exposed to ultraviolet-B (UV-B) radiation were investigated. The findings revealed that UV-B treatment significantly increased flavonoid content in buckwheat sprouts, with 3-day-old sprouts exhibiting a flavonoid content 1.73 times greater than that of the control treatment. UV-B radiation significantly increased the activities of key enzymes involved in flavonoid biosynthesis (phenylalanine ammonia-lyase, 4-coumarate-CoA ligase, cinnamate 4-hydroxylase, and chalcone synthase) and the relative expression levels of the corresponding genes. Although UV-B radiation caused damage to the cell membranes of buckwheat sprouts, promoting increases in hydrogen peroxide and malondialdehyde content and inhibiting the growth of sprouts, importantly, UV-B radiation also significantly increased the activities of catalase, peroxidase, and superoxide dismutase as well as the relative expression levels of the corresponding genes, thus enhancing the antioxidant system of buckwheat sprouts. This enhancement was corroborated by a notable increase in ABTS, DPPH, and FRAP radical scavenging activities in 3-day-old sprouts subjected to UV-B radiation. Additionally, UV-B radiation significantly increased chlorophyll a and chlorophyll b contents in sprouts. These results suggest that UV-B radiation is advantageous for cultivating buckwheat sprouts with increased flavonoid content and enhanced antioxidant capacity. [ABSTRACT FROM AUTHOR]

Anthocyanins in maize (Zea mays L.) kernels determine the plant's color and can enhance its resistance. Selenium (Se) significantly impacts plant growth, development, and secondary metabolic regulation. However, the molecular mechanisms by which Se regulates anthocyanin synthesis in waxy corn remain unclear. This study employed integrated transcriptomic and metabolomic analyses to investigate the mechanisms through which selenium influences anthocyanin synthesis in yellow and purple waxy corn. The results showed that maize varieties with higher anthocyanin content had higher selenium enrichment capacity in their kernels. Under selenium stress, HN2025 exhibited 1,904 more differentially expressed genes (DEGs) and 140 more differential metabolites compared to HN5. The expression levels of anthocyanin synthesis-related genes and transcription factors such as phenylalanine ammonia-lyase, flavonoid 3-hydroxylase (F3H), dihydroflavonol reductase (DFR), chalcone synthase (CHS), cinnamate-4-hydroxylase (C4H), anthocyanin 5,3-O-glucosyltransferases, and anthocyanidin reductase, MYB, and bHLH were strongly induced in HN2025. Metabolomic analysis revealed significant enrichment in anthocyanin biosynthesis, flavonoid and flavonol biosynthesis, glutathione metabolism, phenylalanine biosynthesis, and phenylalanine metabolism under selenium treatment. Three up-regulated PAL genes and one C4H gene were significantly enriched with DAMs in phenylalanine metabolism, phenylpropanoid biosynthesis, flavonoid biosynthesis, and anthocyanin biosynthesis, resulting in significant differences between HN5 and HN2025 in selenium-induced anthocyanin metabolism-related pathways. These findings provide a theoretical basis for understanding the effects of selenium on the molecular regulatory mechanisms of anthocyanin biosynthesis in maize kernels. [ABSTRACT FROM AUTHOR]

Anthocyanin‐enriched Chinese cabbage has health‐enhancing antioxidant properties. Although various regulators of anthocyanin biosynthesis have been identified, the role of individual repressors in this process remains underexplored. This study identifies and characterizes the R2R3‐MYB BrMYB32 in Chinese cabbage (Brassica rapa), which acts as a repressor in anthocyanin biosynthesis. BrMYB32 expression is significantly upregulated under anthocyanin inductive conditions, such as sucrose and high light treatment. Transgenic tobacco plants overexpressing BrMYB32 show decreased anthocyanin levels and downregulation of anthocyanin biosynthesis genes in flowers, highlighting BrMYB32's repressive role. Located in the nucleus, BrMYB32 interacts with the TRANSPARENT TESTA 8 (BrTT8), a basic helix–loop–helix protein, but no interaction was detected with the R2R3‐MYB protein PRODUCTION OF ANTHOCYANIN PIGMENT 1 (BrPAP1). Functional assays in Chinese cabbage cotyledons and tobacco leaves demonstrate that BrMYB32 represses the transcript level of anthocyanin biosynthesis genes, thereby inhibiting pigment accumulation. Promoter activation assays further reveal that BrMYB32 inhibits the transactivation of CHALCONE SYNTHASE and DIHYDROFLAVONOL REDUCTASE through the C1 and C2 motifs. Notably, BrMYB32 expression is induced by BrPAP1, either alone or in co‐expression with BrTT8, and subsequently regulates the expression of these activators. It verifies that BrMYB32 not only interferes with the formation of an active MYB–bHLH–WD40 complex but also downregulates the transcript levels of anthocyanin biosynthesis genes, thereby fine‐tuning anthocyanin biosynthesis. Our findings suggest a model in which anthocyanin biosynthesis in Chinese cabbage is precisely regulated by the interplay between activators and repressors. [ABSTRACT FROM AUTHOR]

Garden dahlias (Dahlia pinnata) are popular for their wide range of color variations, with polychromatic cultivars enhancing their ornamental value. Previous studies on the anthocyanin biosynthetic pathway (ABP) have indicated that the post-transcriptional suppression of the chalcone synthase gene (CHS) is involved in the formation of the white petals of dahlias. To further explore the complex mechanisms underlying polychromatic petal formation, we selected the bicolor cultivar 'LiRen' to identify candidate genetic factors. Through the detection of proanthocyanidin and anthocyanin, it was indicated that the white tips of the petals lacked anthocyanin but accumulated some proanthocyanidin, albeit at significantly lower levels than those at the red bases of the petals. This suggests that the upstream ABP, which involves CHS, is not entirely inactive. Transcription sequencing and quantitative reverse transcription PCR (qRT-PCR) analysis demonstrated that the inactive ABP in the white tips results from the downregulation of ABP structural genes. The low abundance of DpMYB1 appears to be the key factor influencing the lack of strong transcription activation of the structural genes. Additionally, highly upregulated DpSPL9 targeted by the downregulated miR156 in the white tips was identified through qRT-PCR. This suggests that DpSPL9 may act as an anthocyanin depressor to destabilize the MYB-bHLH-WDR complex through interaction with DpMYB1. The findings indicate that the DpMYB1 and miR156-DpSPL9 modules play potential regulatory roles in the formation of bicolor petals. Overall, these results provide new insights into the color patterning of dahlias and will be valuable for further studies regarding the mechanisms underlying polychromatic petal formation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Burdock fructooligosaccharide (BFO) is fructose with a low polymerization degree, which could improve the immunity to pathogens, quality, and stress resistance of vegetables. Still, there are no studies on applying BFO in Chinese cabbage. In this study, the effects of exogenous BFO sprayed with different concentrations (0, 5, 10, 20, 30 g·L−1) on the growth and soluble sugar content of Chinese cabbage seedlings were determined. The result showed that 10 g·L−1 was the appropriate spraying concentration. Based on metabolome analysis, a total of 220 differentially accumulated metabolites (DAMs) were found, among which flavonoid metabolites, glucosinolate metabolites, and soluble sugar-related metabolites were the key metabolites involved in improving the quality of Chinese cabbage caused by BFO. Further combination analysis with transcriptome, trans-cinnamate 4-monooxygenase (CYP73A5), and chalcone synthase 1 (CHS1) were more closely associated with the DAMs of flavonoid biosynthesis. Sulfotransferases 18 (SOT18), Branched-chain amino acid amino transferases 6 (BCAT6), and cytochrome P450 monooxygenase (CYP83A1) were the key genes in glucosinolate biosynthesis. Hexokinase (HxK1), beta-glucosidase 8 (BGL08), invertase 3 (INV3), beta-glucosidase 3B (BGL3B), and sucrose phosphate synthase 1 (SPS1) were significantly upregulated, potentially playing crucial roles in the soluble sugar metabolism. In conclusion, these results provided an understanding of the effects of BFO on the expression of genes and the accumulation of metabolites related to quality formation in Chinese cabbage. [ABSTRACT FROM AUTHOR]

This study aims to investigate the potential mechanism of melatonin (MT) to alleviate the browning of postharvest lotus seeds. 1) The samples were treated with the distilled water, nitric oxide (NO), MT, the inhibitor of nitric oxide synthase (NOS) nomega-nitro-L-arginine methyl ester (L-NAME), the inhibitor of nitrate reductase (NR) tungstate (TUN), and the scavenger of NO carboxy-PTIO (cPTIO), MT + L-NAME, MT + TUN and MT + cPTIO. A systematic analysis was implemented to explore the possible pathway of NO biosynthesis in the lotus seeds. Then, the levels of key enzymes and substances were determined to involve in the NO biosynthesis. The results showed that the endogenous NO content of the MT treated lotus seeds was significantly higher than that of the control. The NOS activity of lotus seeds treated with MT was promoted by 60.36%−71.08% before 3 days of storage. In addition, the MT treatment significantly increased the contents of Larginine and citrulline (P <0.05), which were the key substances in the NO biosynthesis. It was notable that the citrulline content in the MT treated sample was 1.44~1.59 times than those of the controls. By contrast, there was the varying influence of MT treatment on the activity of NR in lotus seeds. There was the beneficial effect of MT on the lotus seeds and the endogenous NO content of the tissue, when the MT was combined with the inhibitor of NR (TUN). However, this beneficial effect disappeared, when the MT was combined with the inhibitor of NOS (L-NAME) for treating the lotus seeds. The endogenous NO content in this treatment was significantly lower than that in the control. Therefore, the exogenous MT treatment was induced the NO biosynthesis in lotus seeds through the pathway of NOS. 2) The potential mechanism of MT was clarified to regulate the browning of lotus seeds. The phenolic compounds were identified in the lotus seeds using a LC20 HPLC (Shimadzu, Japan) system coupled to a TripleTOF® 5600 + quadrupole time-of-flight (QTOF) mass spectrometer equipped with a DuoSpray™ ion source (Sciex, Ontario, Canada). It was found that there were six flavanols and seven phenolic acids in the lotus seeds, and the content of catechin, a type of flavanol, accounts for over 60% of the total phenol content, the following was ellagitannin, a type of phenolic acid, accounts for 7.45% of the total phenol content. It infers that the catechin was the major phenolic compound of lotus seeds. The flavan-3-ol was the most common direct natural substrate of polyphenol oxidase (PPO) in the plant, especially for the catechin and epicatechin. Thereby, the potential mechanism of MT was determined, where the catechin metabolism was regulated to induce the NO production in the following experiments. The results indicated that the activity of PPO in the MT treated lotus seeds was lower by 35.39%−57.36% than those in the control. What’s more, the MT treatment significantly increased the activities of key enzymes, including cinnamate-4-hydroxylase, dihydroflavonol reductase, chalcone synthase, chalcone isomerase and colorless anthocyanin reductase (P <0.05) in the catechin synthetic metabolism. As a result, the content of catechin in the MT treated sample was higher by 15.35%−47.86% than those in the control after 1 days of storage. Whereas, this positive effect was negated, when the MT was combined with LNAME or cPTIO for treating lotus seeds. Therefore, it was concluded that the exogenous MT treatment was induced the biosynthesis of NO through NOS pathway, and then the accumulated NO acted on the catechin metabolism of lotus seeds. The MT treatment was used to inhibit the PPO activity, and then the catechin was suppressed to participate in the enzymatic browning; The biosynthesis of catechin was promoted in this treatment. Consequently, the lotus seeds browning was also alleviated by the MT treatment. These findings can provide the theoretical and technical support to preservation of lotus seeds and the signal transduction between MT and NO. [ABSTRACT FROM AUTHOR]

Objective: To analyze the differences of goji flavonoids from different origins from the perspective of proteomics. Methods: The protein differences of goji berries from different origins were explored by tandem mass tag (TMT) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology in Ningxia, Qinghai and Gansu. Results: A total of 716 differential expressed proteins (DEPs), and a total of four metabolic pathways associated with flavonoids were found in the KEGG pathway, and the phenylpropane biosynthesis pathway was significantly enriched in all three comparison groups (P < 0.05), indicating that the phenylpropane biosynthesis pathway might make a greater contribution to the synthesis of goji flavonoids. Further screening of 24 key differentially expressed proteins related to flavonoid synthesis, it was found that phenylalanine ammonia-lyase (PAL), cinnamic acid 4-hydroxylase (C4H), chalcone isomerase (CHI), chalcone synthase (CHS), etc. might be important DEPs for the biosynthesis of flavonoids in Ningxia, Qinghai and Gansu. Conclusion: The differential proteins related to the biosynthesis of goji flavonoids were identified from the protein level, and the anabolic pathway of goji flavonoids was preliminarily speculated, which laid a foundation for exploring the differences in the biosynthesis of goji flavonoids in different origins. [ABSTRACT FROM AUTHOR]

Ferns represent the second-largest group of vascular plants, yet their genomic resources lag far behind. Here, we present a chromosome-scale genome assembly of Cibotium barometz (L.) J. Sm., a medicinally important fern species. The 3.49 Gb genome, assembled into 66 chromosomes with 99.41% sequence anchorage, revealed an exceptionally high proportion (83.93%) of repetitive elements, dominated by recently expanded LTR retrotransposons. We identified 30,616 protein-coding genes, providing insights into fern-specific gene families. Genomic analyses uncover the evolutionary dynamics of 513 key biosynthetic genes, particularly those involved in terpenoid and flavonoid production. Expression profiling across tissues revealed tissue-specific regulation of these pathways, with notable upregulation of chalcone synthase genes in roots. Our structural analysis of 1-deoxy-d-xylulose-5-phosphate synthase, a key enzyme in terpenoid biosynthesis, demonstrated high conservation across land plants while highlighting fern-specific adaptations. The identification of multiple isoforms for key enzymes points to potential gene-duplication events or the evolution of fern-specific variants. This genome provides a foundation for understanding fern biology, evolution, and the molecular basis of their medicinal properties. It also offers valuable resources for conservation efforts and pharmacological research, paving the way for sustainable utilization of this valuable medicinal plant and advancing our understanding of plant diversity and natural product biosynthesis. [ABSTRACT FROM AUTHOR]

Camellia hainanica is one of the camellia plants distributed in tropical regions, and its regeneration system and genetic transformation are affected by callus browning. However, the underlying mechanism of Camellia hainanica callus browning formation remains largely unknown. To investigate the metabolic basis and molecular mechanism of the callus browning of Camellia hainanica, histological staining, high-throughput metabolomics, and transcriptomic assays were performed on calli with different browning degrees (T1, T2, and T3). The results of histological staining revealed that the brown callus cells had obvious lignification and accumulation of polyphenols. Widely targeted metabolomics revealed 1190 differentially accumulated metabolites (DAMs), with 53 DAMs annotated as phenylpropanoids and flavonoids. Comparative transcriptomics revealed differentially expressed genes (DEGs) of the T2 vs. T1 associated with the biosynthesis and regulation of flavonoids and transcription factors in Camellia hainanica. Among them, forty-four enzyme genes associated with flavonoid biosynthesis were identified, including phenylalaninase (PAL), 4-coumaroyl CoA ligase (4CL), naringenin via flavanone 3-hydroxylase (F3H), flavonol synthase (FLS), Chalcone synthase (CHS), Chalcone isomerase (CHI), hydroxycinnamoyl-CoA shikimate transferase (HCT), Dihydroflavonol reductase (DFR), anthocyanin reductase (LAR), anthocyanin synthetase (ANS), and anthocyanin reductase (ANR). Related transcription factors R2R3-MYB, basic helix-loop-helix (bHLH), and WRKY genes also presented different expression patterns in T2 vs. T1. These results indicate that the browning of calli in Camellia hainanica is regulated at both the transcriptional and metabolic levels. The oxidation of flavonoids and the regulation of related structural genes and transcription factors are crucial decisive factors. This study preliminarily revealed the molecular mechanism of the browning of the callus of Camellia hainanensis, and the results can provide a reference for the anti-browning culture of Camellia hainanica callus. [ABSTRACT FROM AUTHOR]

Jasmonates, including jasmonic acid (JA) and its derivatives such as methyl jasmonate (MeJA) or jasmonly isoleucine (JA-Ile), regulate plant responses to various biotic and abiotic stresses. In this study, we applied exogenous MeJA onto Senna tora leaves subjected to wounding and conducted a transcriptome deep sequencing analysis at 1 (T1), 3 (T3), 6 (T6), and 24 (T24) h after MeJA induction, along with the pretreatment control at 0 h (T0). Out of 18,883 mapped genes, we identified 10,048 differentially expressed genes (DEGs) between the T0 time point and at least one of the four treatment times. We detected the most DEGs at T3, followed by T6, T1, and T24. We observed the upregulation of genes related to JA biosynthesis upon exogenous MeJA application. Similarly, transcript levels of genes related to flavonoid biosynthesis increased after MeJA application and tended to reach their maximum at T6. In agreement, the flavonols kaempferol and quercetin reached their highest accumulation at T24, whereas the levels of the anthraquinones aloe-emodin, emodin, and citreorosein remained constant until T24. This study highlights an increase in flavonoid biosynthesis following both MeJA application and mechanical wounding, whereas no significant influence is observed on anthraquinone biosynthesis. These results provide insights into the distinct regulatory pathways of flavonoid and anthraquinone biosynthesis in response to MeJA and mechanical wounding. [ABSTRACT FROM AUTHOR]

Anthocyanins in maize (Zea mays L.) kernels determine the plant's color and can enhance its resistance. Selenium (Se) significantly impacts plant growth, development, and secondary metabolic regulation. However, the molecular mechanisms by which Se regulates anthocyanin synthesis in waxy corn remain unclear. This study employed integrated transcriptomic and metabolomic analyses to investigate the mechanisms through which selenium influences anthocyanin synthesis in yellow and purple waxy corn. The results showed that maize varieties with higher anthocyanin content had higher selenium enrichment capacity in their kernels. Under selenium stress, HN2025 exhibited 1,904 more differentially expressed genes (DEGs) and 140 more differential metabolites compared to HN5. The expression levels of anthocyanin synthesis-related genes and transcription factors such as phenylalanine ammonia-lyase, flavonoid 3-hydroxylase (F3H), dihydroflavonol reductase (DFR), chalcone synthase (CHS), cinnamate-4-hydroxylase (C4H), anthocyanin 5,3-Oglucosyltransferases, and anthocyanidin reductase, MYB, and bHLH were strongly induced in HN2025. Metabolomic analysis revealed significant enrichment in anthocyanin biosynthesis, flavonoid and flavonol biosynthesis, glutathione metabolism, phenylalanine biosynthesis, and phenylalanine metabolism under selenium treatment. Three up-regulated PAL genes and one C4H gene were significantly enriched with DAMs in phenylalanine metabolism, phenylpropanoid biosynthesis, flavonoid biosynthesis, and anthocyanin biosynthesis, resulting in significant differences between HN5 and HN2025 in selenium-induced anthocyanin metabolism-related pathways. These findings provide a theoretical basis for understanding the effects of selenium on the molecular regulatory mechanisms of anthocyanin biosynthesis in maize kernels. [ABSTRACT FROM AUTHOR]

Gene innovation plays an essential role in trait evolution. Rhizobial symbioses, the most important N2‐fixing agent in agricultural systems that exists mainly in Leguminosae, is one of the most attractive evolution events. However, the gene innovations underlying Leguminosae root nodule symbiosis (RNS) remain largely unknown. Here, we investigated the gene gain event in Leguminosae RNS evolution through comprehensive phylogenomic analyses. We revealed that Leguminosae‐gain genes were acquired by gene duplication and underwent a strong purifying selection. Kyoto Encyclopedia of Genes and Genomes analyses showed that the innovated genes were enriched in flavonoid biosynthesis pathways, particular downstream of chalcone synthase (CHS). Among them, Leguminosae‐gain type Ⅱ chalcone isomerase (CHI) could be further divided into CHI1A and CHI1B clades, which resulted from the products of tandem duplication. Furthermore, the duplicated CHI genes exhibited exon–intron structural divergences evolved through exon/intron gain/loss and insertion/deletion. Knocking down CHI1B significantly reduced nodulation in Glycine max (soybean) and Medicago truncatula; whereas, knocking down its duplication gene CHI1A had no effect on nodulation. Therefore, Leguminosae‐gain type Ⅱ CHI participated in RNS and the duplicated CHI1A and CHI1B genes exhibited RNS functional divergence. This study provides functional insights into Leguminosae‐gain genetic innovation and sub‐functionalization after gene duplication that contribute to the evolution and adaptation of RNS in Leguminosae. [ABSTRACT FROM AUTHOR]

Colletotrichum siamense, a hemibiotrophic pathogen which caused serious strawberry crown rot. Jasmonic acid (JA) is shown to reduce or promote pathogen infection, but the effect of JA on strawberry crown rot is still unknown. Identified the effect and mechanism of JA on strawberry crown rot is the base of resistance induction and genetic improvement for strawberry crown rot. Exogenous methyl jasmonate (MeJA) was tested for its effect for C. siamense causing strawberry crown rot in this study. MeJA significantly increased lesion width and hypha density caused by C. siamense infection in crown. MeJA reprogrammed crown transcriptome, and it induced 1642 significantly differentially expressed genes. In addition, most differentially expressed genes were most enriched in ‘metabolite biosynthetic processes’ and ‘response to stimulus’ by COG enrichment and KOG function classification. Further, KEGG function enrichment showed ‘flavonoid biosynthesis’ vested in ‘metabolite biosynthetic processes’, ‘plant-pathogen interaction’ vested in ‘response to stimulus’ were suppressed by MeJA. qRT-PCR showed expressions of defense genes like heat shock protein, MYB and cellulose synthase A catalytic subunit 8 and structural genes in ‘flavonoid biosynthesis’ were all suppressed. Confirmed with gene expressions, MeJA decreased total flavonoid and down-regulated activities of chalcone synthase and chalcone isomerase. Thus, exogenous MeJA enhanced C. siamense causing crown rot in strawberry by down-regulating defense genes and flavonoids biosynthesis. [ABSTRACT FROM AUTHOR]

To reveal the effect of sucrose concentration on the production of secondary metabolites, a metabolome and transcriptome joint analysis was carried out using callus induced from grape variety Mio Red cambial meristematic cells. We identified 559 metabolites—mainly flavonoids, phenolic acids, and stilbenoids—as differential content metabolites (fold change ≥2 or ≤0.5) in at least one pairwise comparison of treatments with 7.5, 15, or 30 g/L sucrose in the growing media for 15 or 30 days (d). Resveratrol, viniferin, and amurensin contents were highest at 15 d of subculture; piceid, ampelopsin, and pterostilbene had higher contents at 30 d. A transcriptome analysis identified 1310 and 498 (at 15 d) and 1696 and 2211 (at 30 d) differentially expressed genes (DEGs; log2(fold change) ≥ 1, p < 0.05) in 7.5 vs. 15 g/L and 15 vs. 30 g/L sucrose treatments, respectively. In phenylpropane and isoflavone pathways, DEGs encoding cinnamic acid 4-hydroxylase, chalcone synthase, chalcone isomerase, and flavanone 3-hydroxylase were more highly expressed at 15 d than at 30 d, while other DEGs showed different regulation patterns corresponding to sucrose concentrations and cultivation times. For all three sucrose concentrations, the stilbene synthase (STS) gene exhibited significantly higher expression at 15 vs. 30 d, while two resveratrol O-methyltransferase (ROMT) genes related to pterostilbene synthesis showed significantly higher expression at 30 vs. 15 d. In addition, a total of 481 DEGs were annotated as transcription factors in pairwise comparisons; an integrative analysis suggested MYB59, WRKY20, and MADS8 as potential regulators responding to sucrose levels in flavonoid and stilbene biosynthesis in grape callus. Our results provide valuable information for high-efficiency production of flavonoids and stilbenes using grape callus. [ABSTRACT FROM AUTHOR]

Cotton aphids (Aphis gossypii Glover) cause harm by feeding on phloem sap and spreading plant viruses to lily. Understanding the mechanisms by which aphids infest lily plants is crucial for effective aphid management and control. In this study, we investigated the activity of antioxidants, integrated nontargeted metabolomes and transcriptomes of lilies infested by cotton aphids to explore the changes in lily leaves. Overall, the results indicated that the catalase (CAT) activity in the leaves of the lily plants was greater than that in the leaves of the control plants. A comprehensive identification of 604 substances was conducted in the leaves. Furthermore, the differentially abundant metabolite analysis revealed the enrichment of phenylalanine metabolism and α-linolenic acid metabolism. Moreover, 3574 differentially expressed genes (DEGs), whose expression tended to increase, were linked to glutathione metabolism and phenylpropanoid biosynthesis. In addition, the integrated analysis revealed that the defensive response of lily leaves to aphids is manifested through antioxidant reactions, phenylpropane and flavonoid biosynthesis, and α-linolenic acid metabolism. Finally, the key metabolites were CAT, glutathione, coumaric acid, and jasmonic acid, along with the key genes chalcone synthase (CHS), phenylalanine ammonia-lyase (PAL), and 12-oxo-phytodienoic acid reductase (OPR). Accordingly, the findings of this research elucidate the molecular and metabolic reactions of A. gossypii in lily plants, offering valuable insights for developing aphid resistance strategies in lily farming. [ABSTRACT FROM AUTHOR]

Sunburn stress is one of the main environmental stress factors that seriously affects the fruit development and quality of Chinese olive, a tropical and subtropical fruit in south China. Therefore, the understanding of the changes in physiological, biochemical, metabolic, and gene expression in response to sunburn stress is of great significance for the industry and breeding of Chinese olive. In this study, the different stress degrees of Chinese olive fruits, including serious sunburn injury (SSI), mild sunburn injury (MSI), and ordinary (control check, CK) samples, were used to identify the physiological and biochemical changes and explore the differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) by using transcriptomics and metabolomics. Compared with CK, the phenotypes, antioxidant capacity, and antioxidant-related enzyme activities of sunburn stress samples changed significantly. Based on DEG-based KEGG metabolic pathway analysis of transcriptomics, the polyphenol and flavonoid-related pathways, including phenylpropanoid biosynthesis, sesquiterpenoid, and triterpenoid biosynthesis, monoterpene biosynthesis, carotenoid biosynthesis, isoflavonoid biosynthesis, flavonoid biosynthesis, were enriched under sunburn stress of Chinese olive. Meanwhile, 33 differentially accumulated polyphenols and 99 differentially accumulated flavonoids were identified using metabolomics. According to the integration of transcriptome and metabolome, 15 and 8 DEGs were predicted to regulate polyphenol and flavonoid biosynthesis in Chinese olive, including 4-coumarate-CoA ligase (4CL), cinnamoyl-CoA reductase (CCR), cinnamoyl-alcohol dehydrogenase (CAD), chalcone synthase (CHS), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS). Additionally, the content of total polyphenols and flavonoids was found to be significantly increased in MSI and SSI samples compared with CK. Our research suggested that the sunburn stress probably activates the transcription of the structural genes involved in polyphenol and flavonoid biosynthesis in Chinese olive fruits to affect the antioxidant capacity and increase the accumulation of polyphenols and flavonoids, thereby responding to this abiotic stress. [ABSTRACT FROM AUTHOR]

Seagrasses are a paraphyletic group of marine angiosperms and retain certain adaptations from the ancestors of all embryophytes in the transition to terrestrial environments. Among these adaptations is the production of flavonoids, versatile phenylpropanoid secondary metabolites that participate in a variety of stress responses. Certain features, such as catalytic promiscuity and metabolon interactions, allow flavonoid metabolism to expand to produce novel compounds and respond to a variety of stimuli. As marine environments expose seagrasses to a unique set of stresses, these plants display interesting flavonoid profiles, the functions of which are often not completely clear. Flavonoids will likely prove to be effective and versatile agents in combating the new host of stress conditions introduced to marine environments by anthropogenic climate change, which affects marine environments differently from terrestrial ones. These new stresses include increased sulfate levels, changes in salt concentration, changes in herbivore distributions, and ocean acidification, which all involve flavonoids as stress response mechanisms, though the role of flavonoids in combatting these climate change stresses is seldom discussed directly in the literature. Flavonoids can also be used to assess the health of seagrass meadows through an interplay between flavonoid and simple phenolic levels, which may prove to be useful in monitoring the response of seagrasses to climate change. Studies focusing on the genetics of flavonoid metabolism are limited for this group, but the large chalcone synthase gene families in some species may provide an interesting topic of research. Anthocyanins are typically studied separately from other flavonoids. The phenomenon of reddening in certain seagrass species typically focuses on the importance of anthocyanins as a UV-screening mechanism, while the role of anthocyanins in cold stress is discussed less often. Both of these stress response functions would be useful for adaptation to climate change-induced deviations in tidal patterns and emersion. However, ocean warming will likely lead to a decrease in anthocyanin content, which may impact the performance of intertidal seagrasses. This review highlights the importance of flavonoids in angiosperm stress response and adaptation, examines research on flavonoids in seagrasses, and hypothesizes on the importance of flavonoids in these organisms under climate change.

IntroductionSilybum marianum is a medicinal plant that produces silymarin, which has been demonstrated to possess antiviral, anti-neurodegenerative, and anticancer activities. Silybin (A+B) are two major hepatoprotective flavonolignans produced predominantly in S. marianum fruits. Several attempts have been made to increase the synthesis of silymarin, or its primary components, silybin (A+B). Zinc oxide nanoparticles (ZnO-NPs) are considered a highly efficient Zn source widely used to promote crop development and productivity.MethodsIn this study, we aimed to investigate the effects of the foliar application of ZnO-NPs on the growth, yield, photosynthetic pigment content, silybin (A+B) content, and the expression of the chalcone synthase (CHS) gene in S. marianum plants. Different concentrations of ZnO-NPs were administered as foliar sprays to S. marianum plants growing in greenhouse conditions. Furthermore, we evaluated the anti-osteoporotic efficacy of the corresponding fruit extract against dexamethasone (Dex)-induced osteoporosis.Results and discussionFoliar treatment at all ZnO-NP concentrations increased the amounts of bioactive components of silybin (A+B), which enhanced the growth and yield of S. marianum plants while increasing the levels of N, P, K, and Zn in their leaves, roots, and fruits; the levels of photosynthetic pigments in their leaves; and silybin (A+B) content in their fruits, thereby increasing the medicinal value of S. marianum. The highest gains were observed in plants sprayed with the highest ZnO-NP concentration (20.0 mg/L). In addition, gene expression studies revealed that ZnO-NPs stimulated silybin (A+B) production by activating CHS genes. The administration of S. marianum extracts to Dex-administered rats increased osteoblast and bone formation while inhibiting osteoclast and bone resorption, thereby protecting the animals against Dex-induced osteoporosis.

Abstract Background Polyketide synthases (PKSs) are classified into three types based on their enzyme structures. Among them, type III PKSs, catalyzing the iterative condensation of malonyl-coenzyme A (CoA) with a CoA-linked starter molecule, are important synthases of valuable natural products. However, low efficiency and byproducts formation often limit their applications in recombinant overproduction. Results Herein, a rapid growth selection system is designed based on the accumulation and derepression of toxic acyl-CoA starter molecule intermediate products, which could be potentially applicable to most type III polyketides biosynthesis. This approach is validated by engineering both chalcone synthases (CHS) and host cell genome, to improve naringenin productions in Escherichia coli. From directed evolution of key enzyme CHS, beneficial mutant with ~ threefold improvement in capability of naringenin biosynthesis was selected and characterized. From directed genome evolution, effect of thioesterases on CHS catalysis is first discovered, expanding our understanding of byproduct formation mechanism in type III PKSs. Taken together, a whole-cell catalyst producing 1082 mg L−1 naringenin in flask with E value (evaluating product specificity) improved from 50.1% to 96.7% is obtained. Conclusions The growth selection system has greatly contributed to both enhanced activity and discovery of byproduct formation mechanism in CHS. This research provides new insights in the catalytic mechanisms of CHS and sheds light on engineering highly efficient heterologous bio-factories to produce naringenin, and potentially more high-value type III polyketides, with minimized byproducts formation.

Melatonin (Mt) functions as a growth regulator and multifunctional signaling molecule in plants, thereby playing a crucial role in promoting growth and orchestrating protective responses to various abiotic stresses. However, the mechanism whereby exogenous Mt protects Lonicera japonica Thunb. (L. japonica) against salt stress has not been fully elucidated. Therefore, this study aimed to elucidate how exogenous Mt alleviates sodium chloride (NaCl) stress in L. japonica seedlings. Salt-sensitive L. japonica seedlings were treated with an aqueous solution containing 150 mM of NaCl and aqueous solutions containing various concentrations of Mt. The results revealed that treatment of NaCl-stressed L. japonica seedlings with a 60 µM aqueous solution of Mt significantly enhanced vegetative plant growth by scavenging reactive oxygen species and thus reducing oxidative stress. The latter was evidenced by decreases in electrical conductivity and malondialdehyde (MDA) concentrations. Moreover, Mt treatment led to increases in the NaCl-stressed L. japonica seedlings' total chlorophyll content, soluble sugar content, and flavonoid content, demonstrating that Mt treatment improved the seedlings' tolerance of NaCl stress. This was also indicated by the NaCl-stressed L. japonica seedlings exhibiting marked increases in the activities of antioxidant enzymes (superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase) and in photosynthetic functions. Furthermore, Mt treatment of NaCl-stressed L. japonica seedlings increased their expression of phenylalanine ammonia-lyase 1 (PAL1), phenylalanine ammonia-lyase 2 (PAL2), calcium-dependent protein kinase (CPK), cinnamyl alcohol dehydrogenase (CAD), flavanol synthase (FLS), and chalcone synthase (CHS). In conclusion, our results demonstrate that treatment of L. japonica seedlings with a 60 µM aqueous solution of Mt significantly ameliorated the detrimental effects of NaCl stress in the seedlings. Therefore, such treatment has substantial potential for use in safeguarding medicinal plant crops against severe salinity. [ABSTRACT FROM AUTHOR]

SUMMARY: Due to the chelation of phosphorus in the soil, it becomes unavailable for plant growth and development. The mechanisms by which phosphorus‐solubilizing bacteria activate immobilized phosphorus to promote the growth and development of woody plants, as well as the intrinsic molecular mechanisms, are not clear. Through the analysis of microbial communities in the rhizosphere 16S V3–V4 and a homologous gene encoding microbial alkaline phosphomonoesterase (phoD) in phosphate‐efficient (PE) and phosphate‐inefficient apple rootstocks, it was found that PE significantly enriched beneficial rhizobacteria. The best phosphorus‐solubilizing bacteria, Bacillus sp. strain 7DB1 (B2), was isolated, purified, and identified from the rhizosphere soil of PE rootstocks. Incubating with Bacillus B2 into the rhizosphere of apple rootstocks significantly increased the soluble phosphorus and flavonoid content in the rhizosphere soil. Simultaneously, this process stimulates the root development of the rootstocks and enhances plant phosphorus uptake. After root transcriptome sequencing, candidate transcription factor MhMYB15, responsive to Bacillus B2, was identified through heatmap and co‐expression network analysis. Yeast one‐hybrid, electrophoretic mobility shift assay, and LUC assay confirmed that MhMYB15 can directly bind to the promoter regions of downstream functional genes, including chalcone synthase MhCHS2 and phosphate transporter MhPHT1;15. Transgenic experiments with MhMYB15 revealed that RNAi‐MhMYB15 silenced lines failed to induce an increase in flavonoid content and phosphorus levels in the roots under the treatment of Bacillus B2, and plant growth was slower than the control. In conclusion, MhMYB15 actively responds to Bacillus B2, regulating the accumulation of flavonoids and the uptake of phosphorus, thereby influencing plant growth and development. Significance Statement: In the rhizosphere soil of apple rootstocks on different phosphorus‐efficient/inefficient rootstocks, there exist distinct microbial communities. MhMYB15 can respond to the rhizobacterium Bacillus B2, and interact with flavonoids (MhCHS2) and phosphate transport proteins (MhPHT1;15) to promote plant root development and phosphate absorption. [ABSTRACT FROM AUTHOR]

The application of double-stranded RNAs (dsRNAs) to plant surfaces has emerged as a promising tool for manipulating gene expression in plants and pathogens, offering new opportunities for crop improvement. While research has shown the capability of exogenous dsRNAs to silence genes, the full spectrum of their impact, particularly on the intricate network of microRNAs (miRNAs), remains largely unexplored. Here, we show that the exogenous application of chalcone synthase (CHS)-encoding dsRNA to the rosette leaves of Arabidopsis thaliana induced extensive alterations in the miRNA profile, while non-specific bacterial neomycin phosphotransferase II (NPTII) dsRNA had a minimal effect. Two days after treatment, we detected 60 differentially expressed miRNAs among the 428 miRNAs found in the A. thaliana genome. A total of 59 miRNAs were significantly changed after AtCHS-dsRNA treatment compared with water and NPTII-dsRNA, and 1 miRNA was significantly changed after AtCHS-dsRNA and NPTII-dsRNA compared with the water control. A comprehensive functional enrichment analysis revealed 17 major GO categories enriched among the genes potentially targeted by the up- and downregulated miRNAs. These categories included processes such as aromatic compound biosynthesis (a pathway directly related to CHS activity), heterocycle biosynthesis, RNA metabolism and biosynthesis, DNA transcription, and plant development. Several predicted targets of upregulated and downregulated miRNAs, including APETALA2, SCL27, SOD1, GRF1, AGO2, PHB, and PHV, were verified by qRT-PCR. The analysis showed a negative correlation between the expression of miRNAs and the expression of their predicted targets. Thus, exogenous plant gene-specific dsRNAs induce substantial changes in the plant miRNA composition, ultimately affecting the expression of a wide range of genes. These findings have profound implications for our understanding of the effects of exogenously induced RNA interference, which can have broader effects beyond targeted mRNA degradation, affecting the expression of other genes through miRNA regulation. [ABSTRACT FROM AUTHOR]

Naringenin is a plant polyphenol, widely explored due to its interesting biological activities, namely anticancer, antioxidant, and anti-inflammatory. Due to its potential applications and attempt to overcome the industrial demand, there has been an increased interest in its heterologous production. The microbial biosynthetic pathway to produce naringenin is composed of tyrosine ammonia-lyase (TAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI). Herein, we targeted the efficient de novo production of naringenin in Escherichia coli by performing a step-by-step validation and optimization of the pathway. For that purpose, we first started by expressing two TAL genes from different sources in three different E. coli strains. The highest p-coumaric acid production (2.54 g/L) was obtained in the tyrosine-overproducing M-PAR-121 strain carrying TAL from Flavobacterium johnsoniae (FjTAL). Afterwards, this platform strain was used to express different combinations of 4CL and CHS genes from different sources. The highest naringenin chalcone production (560.2 mg/L) was achieved by expressing FjTAL combined with 4CL from Arabidopsis thaliana (At4CL) and CHS from Cucurbita maxima (CmCHS). Finally, different CHIs were tested and validated, and 765.9 mg/L of naringenin was produced by expressing CHI from Medicago sativa (MsCHI) combined with the other previously chosen genes. To our knowledge, this titer corresponds to the highest de novo production of naringenin reported so far in E. coli. Key points: • Best enzyme and strain combination were selected for de novo naringenin production. • After genetic and operational optimizations, 765.9 mg/L of naringenin was produced. • This de novo production is the highest reported so far in E. coli. [ABSTRACT FROM AUTHOR]

Phenylpropanoid metabolism plays an important role in cantaloupe ripening and senescence, but the mechanism of ozone regulation on phenylpropanoid metabolism remains unclear. This study investigated how ozone treatment modulates the levels of secondary metabolites associated with phenylpropanoid metabolism, the related enzyme activities, and gene expression in cantaloupe. Treating cantaloupes with 15 mg/m3 of ozone after precooling can help maintain postharvest hardness. This treatment also enhances the production and accumulation of secondary metabolites, such as total phenols, flavonoids, and lignin. These metabolites are essential components of the phenylpropanoid metabolic pathway, activating enzymes like phenylalanine ammonia‐lyase, cinnamate 4‐hydroxylase, 4CL, chalcone synthase, and chalcone isomerase. The results of the transcriptional expression patterns showed that differential gene expression related to phenylpropanoid metabolism in the peel of ozone‐treated cantaloupes was primarily observed during the middle and late storage stages. In contrast, the pulp exhibited significant differential gene expression mainly during the early storage stage. Furthermore, it was observed that the level of gene expression in the peel was generally higher than that in the pulp. The correlation between the relative amount of gene changes in cantaloupe, activity of selected enzymes, and concentration of secondary metabolites could be accompanied by positive regulation of the phenylpropanoid metabolic pathway. Therefore, ozone stress induction positively enhances the biosynthesis of flavonoids in cantaloupes, leading to an increased accumulation of secondary metabolites. Additionally, it also improves the postharvest storage quality of cantaloupes. [ABSTRACT FROM AUTHOR]

Rhododendron simsii Planchon is an important ornamental species in the northern hemisphere. Flower color is an important objective of Rhododendron breeding programs. However, information on anthocyanin synthesis in R. simsii is limited. In this research, the regulatory mechanism of anthocyanin biosynthesis in R. simsii was performed through the integrated analysis of metabolome and RNA-seq. A total of 805 and 513 metabolites were screened by positive and negative ionization modes, respectively, In total, 79 flavonoids contained seven anthocyanidins, 42 flavanones, 10 flavans, 13 flavones, and seven flavonols. Methylated and glycosylated derivatives took up the most. Differentially accumulated metabolites were mainly involved in "flavone and flavonol biosynthesis", "cyanoamino acid metabolism", "pyrimidine metabolism", and "phenylalanine metabolism" pathways. For flavonoid biosynthesis, different expression of shikimate O-hydroxycinnamoyltransferase, caffeoyl-CoA O-methyltransferase, flavonoid 3′-monooxygenase, flavonol synthase, dihydroflavonol 4-reductase/flavanone 4-reductase, F3′5′H, chalcone synthase, leucoanthocyanidin reductase, and 5-O-(4-coumaroyl)-D-quinate 3′-monooxygenase genes ultimately led to different accumulations of quercetin, myricetin, cyanidin, and eriodictyol. In flavone and flavonol biosynthesis pathway, differential expression of F3′5′H, flavonoid 3′-monooxygenase and flavonol-3-O-glucoside/galactoside glucosyltransferase genes led to the differential accumulation of quercetin, isovitexin, and laricitrin. This research will provide a biochemical basis for further modification of flower color and genetic breeding in R. simsii and related Rhododendron species. [ABSTRACT FROM AUTHOR]

To explore the regulatory mechanism of endogenous hormones in the synthesis of anthocyanins in Anoectochilus roxburghii (Wall.) Lindl (A. roxburghii) under different light intensities, this study used metabolomics and transcriptomics techniques to identify the key genes and transcription factors involved in anthocyanin biosynthesis. We also analyzed the changes in and correlations between plant endogenous hormones and anthocyanin metabolites under different light intensities. The results indicate that light intensity significantly affects the levels of anthocyanin glycosides and endogenous hormones in leaves. A total of 38 anthocyanin-related differential metabolites were identified. Under 75% light transmittance (T3 treatment), the leaves exhibited the highest anthocyanin content and differentially expressed genes such as chalcone synthase (CHS), flavonol synthase (FLS), and flavonoid 3′-monooxygenase (F3′H) exhibited the highest expression levels. Additionally, 13 transcription factors were found to have regulatory relationships with 7 enzyme genes, with 11 possessing cis-elements responsive to plant hormones. The expression of six genes and two transcription factors was validated using qRT-PCR, with the results agreeing with those obtained using RNA sequencing. This study revealed that by modulating endogenous hormones and transcription factors, light intensity plays a pivotal role in regulating anthocyanin glycoside synthesis in A. roxburghii leaves. These findings provide insights into the molecular mechanisms underlying light-induced changes in leaf coloration and contribute to our knowledge of plant secondary metabolite regulation caused by environmental factors. [ABSTRACT FROM AUTHOR]

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Although nitrogen deficiency and sucrose are linked to anthocyanin synthesis, the potential role of sucrose in regulating anthocyanin biosynthesis under low nitrogen conditions (LN) in purple lettuce (Lactuca sativa L.) remains unclear. We found that adding exogenous sucrose enhanced anthocyanin biosynthesis but significantly inhibited lettuce growth at high concentrations. Optimal results were obtained using 1 mmol/L sucrose in a low-nitrogen nutrient solution (LN + T1). Chlorophyll fluorescence imaging indicated that the addition of exogenous sucrose induced mild stress. Meanwhile, the activities of antioxidant enzymes (SOD, CAT, and POD) and antioxidant capacity were both enhanced. The mild stress activated the antioxidant system, thereby promoting the accumulation of anthocyanins induced by exogenous sucrose. LN + T1 (low nitrogen nutrient solution supplemented with 1 mmol/L sucrose) up-regulated enzyme genes in the biosynthetic pathway of anthocyanins, including phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), dihydroflavonol reductase (DFR), flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), flavone synthase II (FNSII), and anthocyanidin synthase (ANS). Additionally, various transcription factors such as AP2/ERF, MYB, bHLH, C2H2, NAC, C2C2, HB, MADS, bZIP, and WRKY were found to be up-regulated. This study elucidates the regulatory mechanism of anthocyanin metabolism in response to the addition of exogenous sucrose under low nitrogen conditions and provides a nutrient solution formula to enhance anthocyanin content in modern, high-quality agricultural cultivation. [ABSTRACT FROM AUTHOR]

4-methyl-5-vinylthiazole (MVT) is a significant volatile of caladium (Caladium bicolor) which produces a very high level of thiamin thiazole synthase (THI4) in male flowers. We explored transcriptomes upregulating MVT using RNA-seq during the six developmental stages of the male flower (Day−10 to Day0) in C. bicolor 'Tapestry'. THI4 was the highest transcript throughout the male flower development. Additionally, the genes showing the high expression associated with floral volatiles of caladium on Day0 were trans-resveratrol di-O-methyltransferase (ROMT), chalcone synthase (CHS), 3-ketoacyl-CoA thiolase 2 (KAT2), and linalool synthase (TPS). These four genes correspond to the following elevated volatiles of caladium: 1,3,5-trimethoxybenzene, MVT, indole, methyl salicylate, and linalool on Day0 compared to Day−10. The upstream THI4 gene was cloned to drive a fluorescent gene (ZsGreen1) in transient and stable transgenic petunia and tobacco plants, showing the gene expression only in the male tissue. The tissue-specific expression of the caladium THI4 promoter could benefit crop production with minimal modification of plants. Investigating transcriptomes associated with caladium fragrance can help provide insight into understanding the regulatory mechanisms of floral volatiles of caladium. [ABSTRACT FROM AUTHOR]

Naturally synthesized secondarymetabolites in plants are considered an important source of drugs, food additives, etc. Among them, research on natural plant medicinal components and their synthesis mechanisms has always been of high concern. We identified a novel medicinal floral crop, Plumbago auriculata L., that can be treated with methyl jasmonate (MeJA) for the rapid or sustainable production of natural bioactives from hairy roots. In the study, we globally analyzed the changes in the accumulation of plumbagin and others in the hairy roots of Plumbago auriculata L. hairy roots (PAHR) 15834 in P. auriculata L. based on 100 mmol/L of MeJA treatment by RNA-seq profiling, and we found that there was a significant increase in the accumulation of plumbagin and saponin before 24 h. To explain the principle of co-accumulation, it showed thatMeJA induced JA signaling and the shikimic acid pathway, and the methylvaleric acid (MVA) pathway was activated downstream subsequently by the Mfuzz and weighted gene coexpression analysis. Under the shared metabolic pathway, the high expression of PAL3 and HMGR promoted the activity of the "gateway enzymes" phenylalanine ammonia lyase (PAL) and 3-hydroxy-3-methylglutaryl CoA reductase (HMGR), which respectively induced the high expression of key reaction enzyme genes, including chalcone synthase (CHS), isopentenyl diphosphate (IPP), and farnesyl pyrophosphate synthase (FPS), that led to the synthesis of plumbagin and saponin. We speculated that large amounts of ketones and/or aldehydes were formed under the action of these characteristic enzymes, ultimately achieving their coaccumulation through polyketone and high-level sugar and amino acid metabolism. The study results provided a theoretical basis for carrying out the factory refinement and biosynthesis of plumbagin and saponins and also provided new ideas for fully exploiting multifunctional agricultural crops and plants and developing new agricultural by-products. [ABSTRACT FROM AUTHOR]

The application of phosphorus-solubilizing bacteria is an effective method for increasing the available phosphorus content and inhibiting wheat uptake of heavy metals. However, further research is needed on the mechanism by which phosphorus-solubilizing bacteria inhibit cadmium (Cd) uptake in wheat roots and its impact on the expression of root-related genes. Here, the effects of strain Klebsiella aerogenes M2 on Cd absorption in wheat and the expression of root-related Cd detoxification and immobilization genes were determined. Compared with the control, strain M2 reduced (64.1–64.6%) Cd uptake by wheat roots. Cd fluorescence staining revealed that strain M2 blocked the entry of exogenous Cd into the root interior and enhanced the immobilization of Cd by cell walls. Forty-seven genes related to Cd detoxification, including genes encoding peroxidase, chalcone synthase, and naringenin 3-dioxygenase, were upregulated in the Cd+M2 treatment. Strain M2 enhanced the Cd resistance and detoxification activity of wheat roots through the regulation of flavonoid biosynthesis and antioxidant enzyme activity. Moreover, strain M2 regulated the expression of genes related to phenylalanine metabolism and the MAPK signaling pathway to enhance Cd immobilization in roots. These results provide a theoretical basis for the use of phosphorus-solubilizing bacteria to remediate Cd-contaminated fields and reduce Cd uptake in wheat. [ABSTRACT FROM AUTHOR]

The purpose of this study was to explore the effects of different LED light quality on the germination of alfalfa seeds and the content of photosynthetic pigments and phenolic compounds in seedlings, so as to provide a basis for further light quality control technology for the sprout vegetable production and industrial production of alfalfa plants. [Methods] The effects of 10 different LED light quality(red light R660 and R630, blue light B450 and B465, green light G520, yellow light Y590, red and blue combination R5B2, R1B1 and R2B5, white light W)on seed germination, seedling growth, anatomy, and phenolic compound synthesis of alfalfa‘WL656'were studied using dark treatment (D) as the control. [Results] Different treatments had no effect on the germination rate, germination potential and germination index of alfalfa seeds. Among monochromatic light treatments, both pure red lights promoted radicle elongation of alfalfa seedlings; B465 treatment significantly increased the stem diameter, epidermal cell thickness, cortical thickness, and individual vessel area of the hypocotyl of seedlings, and B450 significantly increased the diameter of vascular bundles. Different monochromatic light had no significant effect on chlorophyll a content, while the chlorophyll b content was lowest under two different blue light treatments, and the carotenoid content was highest under R660 treatment. Both pure blue lights promoted the accumulation of anthocyanins, total phenols and flavonoids in seedlings, but seedlings contained almost no anthocyanins under G520, Y590 and D treatments. R630 increased the chalcone synthase (CHS) activity in the hypocotyl, and G520 significantly increased the CHS activity in cotyledons. W increased the anthocyanin synthase (ANS) activity in the hypocotyl of seedlings, and Y590 increased the ANS activity in seedlings. Among the three combinations of red and blue light, with the decrease of red light ratio, stem diameter, cortical thickness and CHS activity in hypocotyl decreased, the content of root length, epidermal cell thickness, vascular bundle diameter, and anthocyanins of hypocotyls decreased first and then increased, and the content of photosynthetic pigments and CHS activity in seedling cotyledons increased. The content of total phenols and flavonoids in hypocotyl was highest under R2B5 treatment, and there was no difference in the content of three phenolic compounds in cotyledons. ANS activity in hypocotyl was the highest under R1B1 treatment, and ANS activity in cotyledon was the highest under R5B2 treatment. [Conclusions] Pure red light promoted seedling elongation after seed germination, pure blue light promoted the formation of anthocyanins, flavonoids, and total phenolsin seedlings, and the influence of red and blue combination light on the physiological characteristics of seedlings was more complicated. Taking into account the growth and physiological indicators, this study preliminarily believed that R2B5 treatment was a suitable LED light quality for the growth of alfalfa seedlings as it had moderate growth, high chlorophyll and phenolic content. [ABSTRACT FROM AUTHOR]

Simple Summary: The presence of secondary metabolites like flavonoids, alkaloids, and phenolic compounds in white mulberry (Morus alba) make it an ornamental tree mainly in the Asian subcontinent. These secondary metabolites are synthesized through various biosynthetic pathways. Flavonoids like rutin, quercetin, and kaempferol are produced via the phenylpropanoid and flavonoid biosynthetic pathways. The key enzymes responsible for this pathway include phenylalanine ammonia-lyase, chalcone synthase, and flavonoid hydroxylases. Phenolic compounds like resveratrol and oxyresveratrol are produced via the phenylpropanoid pathway using phenylalanine as a precursor. Presences of these secondary metabolites, like in the case of flavonoids (rutin and quercetin), in the mulberry tree have potential biological activities like antioxidant, anti-inflammatory and neuroprotective effects. Also, they have been found to have great medicinal importance, for example in treating diabetes, obesity and neurodegenerative diseases. Also, phenolic compounds like reservertrol exhibit cardio-protective, anticancer, and anti-aging properties. Traditional natural products have been the focus of research to explore their medicinal properties. One such medicinally important plant is the white mulberry, Morus alba, widely distributed in the Asian subcontinent. It is one of the most cultivated species of mulberry tree and has attracted more focus from researchers because of its abundance in phytochemicals as well as multipurpose uses. The leaves, fruits and other parts of the white mulberry plant act as a source of valuable bioactive compounds like flavonoids, phenolic acids, terpenoids and alkaloids. These secondary metabolites have manifold healthy uses as they possess antioxidant, anti-inflammatory, antidiabetic, neutrotrophic, and anticancer properties. Despite the increasing scientific interest in this plant, there are very few reviews that highlight the phytochemistry and biological potential of white mulberry for biomedical research. To this end, this review elaborates the phytochemistry, biosynthetic pathways and pharmacological activities of the glycoside flavonoids of Morus alba. A comprehensive analysis of the available literature indicates that Morus alba could emerge as a promising natural agent to combat diverse conditions including diabetes, cancer, inflammation and infectious diseases. To achieve such important objectives, it is crucial to elucidate the biosynthesis and regulation mechanisms of the bioactive compounds in white mulberry as well as the multifaceted pharmacological effects attributed to this plant resource. The present review paper is intended to present a summary of existing scientific data and a guide for further research in the phytochemistry and pharmacology of white mulberry. Further, a biosynthetic pathway analysis of the glycoside flavonoid in mulberry is also given. Lastly, we discuss the pros and cons of the current research to ensure the prudent and effective therapeutic value of mulberry for promoting human and animal health. [ABSTRACT FROM AUTHOR]

Background: Polyketide synthases (PKSs) are classified into three types based on their enzyme structures. Among them, type III PKSs, catalyzing the iterative condensation of malonyl-coenzyme A (CoA) with a CoA-linked starter molecule, are important synthases of valuable natural products. However, low efficiency and byproducts formation often limit their applications in recombinant overproduction. Results: Herein, a rapid growth selection system is designed based on the accumulation and derepression of toxic acyl-CoA starter molecule intermediate products, which could be potentially applicable to most type III polyketides biosynthesis. This approach is validated by engineering both chalcone synthases (CHS) and host cell genome, to improve naringenin productions in Escherichia coli. From directed evolution of key enzyme CHS, beneficial mutant with ~ threefold improvement in capability of naringenin biosynthesis was selected and characterized. From directed genome evolution, effect of thioesterases on CHS catalysis is first discovered, expanding our understanding of byproduct formation mechanism in type III PKSs. Taken together, a whole-cell catalyst producing 1082 mg L−1 naringenin in flask with E value (evaluating product specificity) improved from 50.1% to 96.7% is obtained. Conclusions: The growth selection system has greatly contributed to both enhanced activity and discovery of byproduct formation mechanism in CHS. This research provides new insights in the catalytic mechanisms of CHS and sheds light on engineering highly efficient heterologous bio-factories to produce naringenin, and potentially more high-value type III polyketides, with minimized byproducts formation. [ABSTRACT FROM AUTHOR]

The flavonoids in citrus fruits are crucial physiological regulators and natural bioactive products of high pharmaceutical value. Melatonin is a pleiotropic hormone that can regulate plant morphogenesis and stress resistance and alter the accumulation of flavonoids in these processes. However, the direct effect of melatonin on citrus flavonoids remains unclear. In this study, nontargeted metabolomics and transcriptomics were utilized to reveal how exogenous melatonin affects flavonoid biosynthesis in "Bingtangcheng" citrus fruits. The melatonin treatment at 0.1 mmol L−1 significantly increased the contents of seven polymethoxylated flavones (PMFs) and up-regulated a series of flavonoid pathway genes, including 4CL (4-coumaroyl CoA ligase), FNS (flavone synthase), and FHs (flavonoid hydroxylases). Meanwhile, CHS (chalcone synthase) was down-regulated, causing a decrease in the content of most flavonoid glycosides. Pearson correlation analysis obtained 21 transcription factors co-expressed with differentially accumulated flavonoids, among which the AP2/EREBP members were the most numerous. Additionally, circadian rhythm and photosynthesis pathways were enriched in the DEG (differentially expressed gene) analysis, suggesting that melatonin might also mediate changes in the flavonoid biosynthesis pathway by affecting the fruit's circadian rhythm. These results provide valuable information for further exploration of the molecular mechanisms through which melatonin regulates citrus fruit metabolism. [ABSTRACT FROM AUTHOR]

The current study sought to determine the effects of Colpomenia sinuosa and Sargassum linifolium aqueous extracts on non-enzymatic and enzymatic antioxidants in Spinacia oleracea L. leaf extract. Glutathione synthase (GSS, EC: 6.3.2.3) and -glutamyl cysteine synthetase (-GCS, EC: 6.3.2.2) are involved in the glutathione biosynthesis process. Treatment of S. oleracea with seaweed extracts increased the level of reduced glutathione (GSH), oxidized glutathione (GSSG), and total glutathione at lower concentrations. Total phenols and total flavonoids in S. oleracea leaves accumulated more rapidly after treatment with seaweed extracts. The activity of the enzyme’s phenylalanine ammonia lyase (PAL, EC: 4.3.1.25), chalcone synthase (CHS, EC: 2.3. 1.74), and chalcone isomerase (CHI, EC: 5.5.1.6) involved in the production of phenylpropanoid and flavonoid in S. oleracea leaves rose in a dose-dependent manner. Glutathione reductase (GR, EC 1.6.4.2), glutathione peroxidase (GPX, EC: 1.11.1.9), and glutathione-S-transferase (GST, EC: 2.5.1.18) are examples of antioxidant enzymes whose activity were also elevated by the treatment. In comparison to untreated plants, the treated plants' S. oleracea leaf extract significantly reduced superoxide anion, 1, 1-diphenyl-2-picrylhydrazyl (DPPH), and hydroxyl radicals. In light of this, the current findings imply that the use of seaweed extracts was found to boost the activity of the antioxidant system in Spinacea oleracea [ABSTRACT FROM AUTHOR]

Chalcone synthase (CHS) and chalcone isomerase (CHI) catalyze the first two committed steps of the flavonoid pathway that plays a pivotal role in the growth and reproduction of land plants, including UV protection, pigmentation, symbiotic nitrogen fixation, and pathogen resistance. Based on the obtained X-ray crystal structures of CHS, CHI, and chalcone isomerase-like protein (CHIL) from the same monocotyledon, Panicum virgatum, along with the results of the steady-state kinetics, spectroscopic/thermodynamic analyses, intermolecular interactions, and their effect on each catalytic step are proposed. In addition, PvCHI's unique activity for both naringenin chalcone and isoliquiritigenin was analyzed, and the observed hierarchical activity for those type-I and -II substrates was explained with the intrinsic characteristics of the enzyme and two substrates. The structure of PvCHS complexed with naringenin supports uncompetitive inhibition. PvCHS displays intrinsic catalytic promiscuity, evident from the formation of p-coumaroyltriacetic acid lactone (CTAL) in addition to naringenin chalcone. In the presence of PvCHIL, conversion of p-coumaroyl-CoA to naringenin through PvCHS and PvCHI displayed ~400-fold increased Vmax with reduced formation of CTAL by 70%. Supporting this model, molecular docking, ITC (Isothermal Titration Calorimetry), and FRET (Fluorescence Resonance Energy Transfer) indicated that both PvCHI and PvCHIL interact with PvCHS in a non-competitive manner, indicating the plausible allosteric effect of naringenin on CHS. Significantly, the presence of naringenin increased the affinity between PvCHS and PvCHIL, whereas naringenin chalcone decreased the affinity, indicating a plausible feedback mechanism to minimize spontaneous incorrect stereoisomers. These are the first findings from a three-body system from the same species, indicating the importance of the macromolecular assembly of CHS-CHI-CHIL in determining the amount and type of flavonoids produced in plant cells. [ABSTRACT FROM AUTHOR]