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Ultraviolet-B (UV-B) radiation is a major abiotic stress that dragon trees are exposed to during their growth and development; however, it is also an environmental signal perceived by plants that affects the flavonoid pathway. Previous studies have demonstrated that amounts of flavonoids are contained in dragon tree resin, otherwise known as dragon's blood. However, the traits and mechanism involved in the UV-B-mediated increase in flavonoids in dragon trees are still unknown. Here, we studied the response of Dracaena cambodiana under full solar UV-B radiation. The results showed that the contents of total flavonoids in D. cambodiana significantly increased after UV-B radiation exposure. Then, the transcriptome was used for determining the interactive mechanism of flavonoid accumulation and UV-B stress. Differential expression analyses identified 34 differentially expressed genes (DEGs) involved in flavonoid synthesis; specifically, 24 of the identified DEGs were significantly up-regulated after UV-B radiation exposure. In addition, 57 DEGs involved in Ca2+/kinase sensors, 58 DEGs involved in ROS scavenging and the plant hormone pathway, and 116 DEGs transcription factors in 5 families were further identified and analyzed. Finally, we deduced the potential mechanism of UV-B-promoting flavonoid formation to neutralize ROS damage derived from UV-B radiation in D. cambodiana based on the gene co-expression network and previous studies from other plants. Considering that wild dragon tree populations are currently highly threatened by anthropogenic and natural stressors, the interactive studies between D. cambodiana plants and UV-B radiation provide valuable information toward understanding the mechanism of dragon's blood formation and help us reveal the evolution of D. cambodiana, with the eventual goal of aiding in the global conservation of this precious biological resource. [ABSTRACT FROM AUTHOR]

Methyl jasmonate (MJ) has potential to regulate fruit ripening and quality. 'Yoho' and 'Jiro' persimmons were sprayed with MJ (0, 2, 4, and 6 mM), four weeks before anticipated harvest to evaluate its effects on fruit colour and bioactive compounds. Preharvest MJ application significantly improved fruit colour with increased a*, b*, chroma, and colour index. The MJ 6 mM application had significantly enhanced soluble solids content (SSC), reduced total chlorophyll content in peel and pulp, and soluble and total tannins in persimmons. MJ treatments exhibited higher contents of total phenolics, flavonoids, carotenoids, and antioxidant activities. Additionally, MJ treatments enhanced the activities of shikimate dehydrogenase (SKDH), phenylalanine ammonia-lyase (PAL), catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and lipoxygenase (LOX) enzymes. Overall, pre-harvest MJ application at 6 mM four weeks before anticipated harvest could be useful for advancing colour and improving bioactive compounds in 'Yoho' and 'Jiro' persimmons., 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 Ltd.. All rights reserved.)

The sliced and dried hawthorn berries are easily infested by insects during storage. This study aimed to determine the effect of insect infestation on the quality of hawthorn berries and assess the change at metabolite level by analyzing physicochemical property and metabolomics profiling. A total of 184 shared differential metabolites were obtained, mainly including flavonoids, fatty acids, carboxylic acids and derivatives, and nitrogenous compounds. Through receiver operating characteristic curve assessment, 9 significant differential markers were screened out to distinguish insect infestation of hawthorn berries. Correlation analysis showed that the color, total organic acids, total phenolics, and total flavonoids were effective indicators for quality evaluation of insect infestation, and uric acid and hippuric acid can serve as biomarkers for the quality deterioration of hawthorn berries during storage. This study demonstrated that insect infestation could decrease the quality of hawthorn berries from macro and micro perspectives., 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 Ltd. All rights reserved.)

Background: Dendrobium catenatum is a perennial herb of the genus Dendrobium orchidaceae. It has been known as "Golden Grass, Soft Gold" since ancient times with effects of strengthening the body, benefiting the stomach, generating body fluid, nourishing Yin and clearing internal heat. The flowers of D. catenatum have anti-oxidation, immune regulation and other biological activities. The composition analysis of flowers showed that flavonoid glycosides were significantly accumulated in floral tissue. However, in the flowers of D. catenatum, there was only one case of the UDP-glycosyltransferase (UGT) responsible for the glycosylation of flavonoids has been reported., Result: In this study, a new UGT (named UGT708S6) was cloned from D. catenatum flowers rich in O-glycosides and C-glycosides, and its function and biochemical properties were characterized. Through homology comparison and molecular docking, we identified the key amino acid residues affecting the catalytic function of UGT708S6. The glycosyltransferase UGT708S6 was characterized and demonstrated C-glycosyltransferase (CGT) activity in vitro assay using phloretin and 2-hydroxynaringenin as sugar acceptors. The catalytic promiscuity assay revealed that UGT708S6 has a clear sugar donor preference, and displayed O-glycosyltransferase (OGT) activity towards luteolin, naringenin and liquiritigenin. Furthermore, the catalytic characteristics of UGT708S6 were explored, shedding light on the structural basis of substrate promiscuity and the catalytic mechanism involved in the formation of flavonoid C-glycosides. R271 was a key amino acid residue site that sustained the catalytic reaction. The smaller binding pocket resulted in the production of new O-glycosides and the reduction of C-glycosides. This highlighted the importance of the binding pocket in determining whether C-glycosides or O-glycosides were produced., Conclusions: The findings suggest that UGT708S6 holds promise as a new glycosyltransferase for synthesizing flavonoid glycosides and offer valuable insights for further understanding the catalytic mechanisms of flavonoid glycosyltransferases., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Flavonoids are a structurally diverse group of secondary metabolites, predominantly produced by plants, which include a range of compounds with pharmacological importance. Pinocembrin is a key branch point intermediate in the biosynthesis of a wide range of flavonoid subclasses. However, replicating the biosynthesis of these structurally diverse molecules in heterologous microbial cell factories has encountered challenges, in particular the modest pinocembrin titres achieved to date. In this study, we combined genome engineering and enzyme candidate screening to significantly enhance the production of pinocembrin and its derivatives, including chrysin, pinostrobin, pinobanksin, and galangin, in Escherichia coli., Results: By implementing a combination of established strain engineering strategies aimed at enhancing the supply of the building blocks phenylalanine and malonyl-CoA, we constructed an E. coli chassis capable of accumulating 353 ± 19 mg/L pinocembrin from glycerol, without the need for precursor supplementation or the fatty acid biosynthesis inhibitor cerulenin. This chassis was subsequently employed for the production of chrysin, pinostrobin, pinobanksin, and galangin. Through an enzyme candidate screening process involving eight type-1 and five type-2 flavone synthases (FNS), we identified Petroselinum crispum FNSI as the top candidate, producing 82 ± 5 mg/L chrysin. Similarly, from a panel of five flavonoid 7-O-methyltransferases (7-OMT), we found pinocembrin 7-OMT from Eucalyptus nitida to yield 153 ± 10 mg/L pinostrobin. To produce pinobanksin, we screened seven enzyme candidates exhibiting flavanone 3-hydroxylase (F3H) or F3H/flavonol synthase (FLS) activity, with the bifunctional F3H/FLS enzyme from Glycine max being the top performer, achieving a pinobanksin titre of 12.6 ± 1.8 mg/L. Lastly, by utilising a combinatorial library of plasmids encoding G. max F3H and Citrus unshiu FLS, we obtained a maximum galangin titre of 18.2 ± 5.3 mg/L., Conclusion: Through the integration of microbial chassis engineering and screening of enzyme candidates, we considerably increased the production levels of microbially synthesised pinocembrin, chrysin, pinostrobin, pinobanksin, and galangin. With the introduction of additional chassis modifications geared towards improving cofactor supply and regeneration, as well as alleviating potential toxic effects of intermediates and end products, we anticipate further enhancements in the yields of these pinocembrin derivatives, potentially enabling greater diversification in microbial hosts., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Nitrogen fertilization in broccoli is very important for plant growth and productivity. However, it is known that the nitrogen source of the nitrogen fertilizer is also effective on quality parameters. In this study we investigated the effects of different nitrogen sources (ammonium sulphate and calcium nitrate) on bioactive compounds and leaf nutrients of broccoli cultivars (Monet, Italian and Karadede)., Results: The highest total phenolic content was 0.24 mg g - 1 fw in cultivar Monet. Nitrate-based fertilizer treatments provided higher total phenolic (0.28 mg g - 1 fw) and flavonoid (0.56 mg g 1 fw) contents and FRAP (0.24 mmol g - 1 fw) values compared to ammonium-based fertilizer treatments. On the other hand, DPPH (0.77 mmol g - 1 fw) values were higher in the ammonium nitrogen treatments. Among the cultivars, Monet had the highest total phenolics, flavonoids and FRAP values. When leaf nutrient concentrations were analyzed, ammonium sulfate application resulted in higher sulfur (0.86%), zinc (20.72 mg kg - 1 ) and manganese (28.80 mg kg - 1 ) concentrations compared to calcium nitrate. Calcium nitrate treatment provided the highest calcium (2.09%) and iron (37.50 mg kg - 1 ) concentration in Monet cultivar. Phosphorus (0.31-0.35%) and magnesium (0.073-0.084%) concentrations differed among cultivars., Conclusions: In conclusion, the choice of nitrogen source and cultivar in broccoli cultivation plays an important role in the bioactive compound profile and nutrient composition of the broccoli crop. The results provide useful information for the improvement of broccoli quality., Competing Interests: Declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Ajuga turkestanica preparations are used as anti-aging cosmeceuticals and for medicinal purposes. Herein we describe the characterization and quantification of its metabolites in different organs using UHPLC-MS and NMR spectroscopy. A total of 51 compounds belonging to various phytochemical classes (11 flavonoids, 10 ecdysteroids, 9 diterpenes, 6 fatty acids, 5 iridoids, 3 phenylpropanoids, 3 sugars, 2 phenolics, 1 coumarin, 1 triterpene) were annotated and tentatively identified by UHPLC-ESI-QqTOF-MS/MS of methanolic extracts obtained separately from the organs. 1D and 2D NMR spectroscopy independently confirmed the identity of six major compounds. The abundances of these main constituents in flowers, fruits, leaves, roots, seeds, and stems were compared and quantified using 1 H NMR. The results showed that 8-O-acetylharpagide, 20-hydroxyecdysone (ecdysterone) and ajugachin B were the most abundant constituents in the species. The two major compounds, 8-O-acetylharpagide and 20-hydroxyecdysone, were chosen as the markers for the quality assessment of A. turkestanica material. The methanolic extract of the aerial parts of A. turkestanica showed no noteworthy anthelmintic (antihelmintic), antifungal, or cytotoxic effect in in vitro assays., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

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.)

With the developing resistance to traditional antiparasitic medications, the purpose of this study was to efficiently develop a series of six noble flavanoidal tetrazinane-6'-one derivatives by a one-pot reaction pathway. FT-IR, 1 HNMR, 13 CNMR, and Mass spectra were employed for the structural elucidation of the synthesized compounds (7-12). Clinostomum complanatum, a parasite infection model that has been well-established, demonstrated that all the synthesized compounds are potent antiparasitic agents. DNA is the main target for various medicinal compounds. As a result, thestudy of how small molecules attach to DNA has received a lot of attention. In the present study, we have performed various biophysical techniques to determine the mode of binding of synthesized compounds (7-12) with calf thymus DNA (ct-DNA). It was observed from the UV-visible absorbance and fluorescence spectra that all synthesized compounds (7-12) form complexes with the ct-DNA. The value of binding constant (K b ) was obtained to be in the range of 4.36---24.50 × 10 3 M - 1 at 298 K. Competitive displacement assay with ethidium bromide (EB), CD spectral analysis, viscosity measurements, and in silico molecular docking confirmed that ligands (7-12) incorporate with ct-DNA through groove binding only. Molecular docking studies were performed for all synthesized compounds with the calf thymus DNA and it was found that all the newly synthesized compounds strongly bind with the chain B of DNA in the minor groove with the value of binding energy in the range of -8.54 to -9.04 kcal per mole and several hydrogen bonding interactions., 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.)

Kombucha was fermented by the pure cultured tea fungus, and the changes of functional compounds and their transformation were explored. After fermentation, the contents of total polyphenols, total flavonoids, quercetin, kaempferol and catechins respectively enhanced by 77.14%, 69.23%, 89.11%, 70.32% and 45.77% compared with the control, while flavonol glycosides reduced by 38.98%. The bioavailability of polyphenols and flavonoids respectively increased by 29.52% and 740.6%, and DPPH and ABTS respectively increased by 43.81% and 35.08% compared with the control. Correlation analysis showed that microorganisms and the antioxidant activity were highly positive correlation with total polyphenols, total flavonoids, EGC, EC, EGCG, ECG, quercetin and kaempferol, and negative correlation with kaempferol-3-glucoside. The constructed models confirmed that organic acids were more likely to damage the structure of tea leaves, and enzymes (polygalacturonidase and tannase) and solvents (acids, alcohols and esters) had a synergistic effect on the biotransformation of functional compounds., Competing Interests: Declaration of competing interest The authors confirm that they have no conflicts of interest with respect to the work described in this manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Bergenia purpurascens is an important medicinal, edible and ornamental plant. The lack of omics information hinders the study of its metabolic pathways and related genes. In order to investigate candidate genes and pathways involved in secondary metabolism in B. purpurascens, roots, stems and leaves of B. purpurascens were subjected to metabolomic and transcriptomic analyses in this study. A total of 351 differentially accumulated secondary metabolites were identified. We identified 120 candidate genes involved in phenylpropanoid and flavonoid biosynthesis pathway, from which 29 key candidate genes were obtained by WGCNA. Five UDP-Glycosyltransferases and four O-methyltransferases were suggested to be the candidate enzymes involved in synthetic pathway from gallic acid to bergenin by correlation analysis between transcriptional and metabolic levels and phylogenetic analysis. This study provides data resources and new insights for further studies on the biosynthesis of major active components in B. purpurascens., Competing Interests: Declarations Ethics approval and consent to participate The B. purpuraceus we purchased originated from Wenchuan Botanical Garden, Wenchuan County, Sichuan Province, a private botanical garden. Consent for publication Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Understanding the influence of light spectra on plant growth and antioxidant activities is crucial for optimizing cultivation practices and enhancing crop quality. In this study, we investigated the effects of different light treatments on growth parameters and antioxidant activities in five plant species: peppermint, Thai basil, cumin, lemon basil, and green holy basil. Our results revealed distinct responses to varying light spectra, with green light consistently promoting taller plant heights across all species. Additionally, blue light induced notable increases in plant width for certain species. Analysis of antioxidant activities demonstrated dynamic fluctuations in Total Phenolic Content (TPC) and Flavonoid Content (TFC) among different light treatments and plant species. While white and red light generally promoted higher TPC levels, blue light unexpectedly exhibited the highest TPC levels at specific time points. Moreover, investigation into DPPH Radical Scavenging activity revealed diverse temporal responses to light spectra, with blue light demonstrating exceptional activity at early stages and white and red light showing heightened activity at later time points. These findings underscore the importance of tailored light regimes in optimizing growth parameters and enhancing antioxidant activities in cultivated plants, thereby offering promising avenues for sustainable agriculture and food production practices., Competing Interests: Declarations Competing interests The authors declare no competing interests. Research involving plants The present study complied with the international and national guidelines for the use of experimental plant material., (© 2024. The Author(s).)

Revealing the interaction mechanism of proteins with bioactive molecules and the location of their binding pockets is crucial for predicting the structure-function relationship of proteins in drug discovery and design. Despite some published papers on the interaction of β-casein with small bioactive molecules, the ambiguity of the location and constituent amino acids of β-casein binding pockets prompted us to identify them by in silico simulation of its interaction with three polyphenols, chrysin, apigenin, and luteolin. Molecular docking revealed that the primary β-casein binding pocket for chrysin consists of five nonpolar amino acids (Leu73, Phe77, Pro80, Ile89, and Pro196), three polar neutral amino acids (Ser137, Gln138, and Gln197), and two polar charged amino acids (Glu136, and Arg198). For β-casein/apigenin and β-casein/luteolin complexes, Asn83 also contributes to forming the pocket. Molecular dynamics provided more details, such as the relative contribution of determinative amino acids and the role of various forces. For example, we found that Glu210, Glu132, and Glu35 are the most destructive residues in the binding of chrysin, apigenin, and luteolin to β-casein, respectively. Also, we observed that hydrophobic forces mainly stabilize β-casein/chrysin and β-casein/apigenin, and polar solvation (including hydrogen bonds) stabilizes β-casein/luteolin, all by spontaneous processes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Amin Sahraei and Reza Sahraei report financial support was provided by Iran National Science Foundation (INSF). If there are other authors, they 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 Inc. All rights reserved.)

Akkermansia muciniphila ( A. muciniphila ) possesses health-promoting properties. Nevertheless, A. muciniphila enrichment remains a challenging endeavor. Quercetin-3- O -rutinose-7- O -α-l-rhamnoside (QRR), a flavonoid found in lychee pulp, has a unique double-substituted glycosylated structure, requiring a specific intestinal microbiota for effective metabolism. Here, QRR was fermented using a coculture of Bacteroides uniformis and A. muciniphila , and the interactions between the two were elucidated in terms of QRR regulation of microbial growth changes and metabolic properties. The results demonstrated that QRR effectively promoted the proliferation of A. muciniphila based on the metabolic action of B. uniformis in vitro, which was evidenced by a notable increase in the number of viable bacteria. Furthermore, the coculture sample exhibited a significant increase in SCFAs. Qualitative analysis of metabolites by UPLC-ESI-Triple-TOF-MS/MS showed that B. uniformis could release sugars on QRR to produce quercetin-3- O -glucoside-7- O -α-rhamnoside and further quercetin. In the coculture and B. uniformis culture, quercetin was converted to taxifolin, which was identified as a crucial intermediate in the metabolism of QRR. Notably, the metabolite kaempferol was only detected in the coculture. The present study reveals the interaction between QRR and the coculture of A. muciniphila and B. uniformis , providing a practical basis for the potential prebiotic value of QRR.

Bioactive compounds play an increasingly prominent role in breeding functional and nutritive fruit crops such as citrus. However, the genomic and metabolic bases for the selection and differentiation underlying bioactive compound variations in citrus remain poorly understood. In this study, we constructed a species-level variation atlas of genomes and metabolomes using 299 citrus accessions. A total of 19 829 significant SNPs were targeted to 653 annotated metabolites, among which multiple significant signals were identified for secondary metabolites, especially flavonoids. Significant differential accumulation of bioactive compounds in the phenylpropane pathway, mainly flavonoids and coumarins, was unveiled across ancestral citrus species during differentiation, which is likely associated with the divergent haplotype distribution and/or expression profiles of relevant genes, including p-coumaroyl coenzyme A 2'-hydroxylases, flavone synthases, cytochrome P450 enzymes, prenyltransferases, and uridine diphosphate glycosyltransferases. Moreover, we systematically evaluated the beneficial bioactivities such as the antioxidant and anticancer capacities of 219 citrus varieties, and identified robust associations between distinct bioactivities and specific metabolites. Collectively, these findings provide citrus breeding options for enrichment of beneficial flavonoids and avoidance of potential risk of coumarins. Our study will accelerate the application of genomic and metabolic engineering strategies in developing modern healthy citrus cultivars., (Copyright © 2024 The Author. Published by Elsevier Inc. All rights reserved.)

Isatis indigotica Fort. is a famous medicinal plant that is also used as a natural dye and functional vegetable. The characteristics of the I. indigotica fruit during development are largely unknown, information that is essential for the exploitation and seedlings cultivation of I. indigotica. In this study, the biochemical, metabolite characteristics and gene expression profiling of I. indigotica at four developmental stages were investigated. A total of 428 metabolites were detected and categorized into 17 categories. High contents of anthocyanins, especially cyanidin 3-glucoside, might contribute to the purple colouration of I. indigotica fruits. Moreover, dozens of flavonoid components, including taxifolin, quercetin, astragalin and isovitexin 2″-O-beta-D-glucoside, and several other active components were also up-regulated in mature fruits. The abundance of antioxidants might endow a significantly stronger antioxidant activity of mature I. indigotica fruits compared to many other reported species. Enrichment analyses revealed that flavonoid and anthocyanin biosynthesis genes were mostly enriched in up-regulated gene sets during fruit development. The up-regulated structural genes, including IiCHS, IiCHI, IiF3H, IiDFR, IiANS, IiFLS, IiUGT, and transcription factors such as IiMYBs, IibHLHs and IiNACs were identified as candidate regulators of flavonoid and anthocyanin biosynthetic pathway. Furthermore, biosynthesis of amino acids was enriched in all pairwise comparisons of metabolites in fruits at four developmental stages. The differential accumulation of amino acids might result from the differentially expressed genes involved in amino acid biosynthesis. Taken together, these findings provide a comprehensive understanding of metabolite profiling and gene expression patterns in I. indigotica fruit during maturity, which is useful for pharmaceutical extractions and seedling cultivation of I. indigotica., (© 2024 Scandinavian Plant Physiology Society.)

Polygonatum rhizome is a traditional Chinese medicine of the same origin as food and medicine, and it has high economic value and social benefits. To screen the excellent germplasm resources of Polygonatum kingianum ( P. kingianum ) and clarify the nutritional and medicinal value of the rhizome of P. kingianum , we used widely targeted metabolomics to analyze the traits and metabolomics of rhizomes of different germplasms of P. kingianum from different growth years. The results showed that different germplasms and growth years of P. kingianum were rich in different nutritional and medicinal components. Among them, Polygonatum kingianum 'Linyun 1' rhizome (PWR) was richer in amino acids and derivatives, alkaloids, and phenolic acids, while Polygonatum kingianum rhizome (PRR) was richer in flavonoids, organic acids, and phenolic acids. Most of the differential compounds were mainly enriched in PRR when the growth year was one, and PWR had a greater variety and higher content of differential compounds in the third year, which also reflected the advantages of Polygonatum kingianum 'Linyun 1' ( P. kingianum 'Linyun 1') as an excellent new variety of P. kingianum . The Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analysis showed that in P. kingianum with the same age and different germplasms, the significantly enriched metabolic pathway was more active in biosynthesis in PWR. In the same germplasm of P. kingianum from different years, the metabolites involved in PRR were mainly the highest in one-year-old P. kingianum (PR-1) or three-year-old P. kingianum (PR-3), and the metabolites involved in PWR were mainly the highest in three-year-old P. kingianum 'Linyun 1' (PW-3). The above results showed that the three-year-old PWR had more advantages based on chemical substances. Therefore, this study provided a new theoretical reference for the development of P. kingianum products and the breeding of new varieties.

Saffron stigma, derived from Crocus sativus L., has long been revered in global traditional medicine and continues to hold significant market value. However, despite the extensive focus on saffron stigma, the therapeutic potential of other floral components remains underexplored, primarily due to limited insights into their complex molecular architectures and chemical diversity. To address this gap, we performed a comprehensive metabolomic analysis of various floral organs utilizing advanced analytical platforms, including GC-MS and UPLC-MS/MS. This in-depth profiling revealed a diverse array of 248 metabolites, encompassing amino acids, sugar derivatives, fatty acids, flavonoids, vitamins, polyamines, organic acids, and a broad spectrum of secondary metabolites. Distinct correlation patterns among these metabolites were identified through PCA and PLS-DA, highlighting unique metabolomic signatures inherent to each floral organ. We further integrated these metabolomic findings with our transcriptomic data, enabling a detailed understanding of the molecular and metabolic variations across different floral organs. The pronounced abundance of differentially expressed genes and metabolites in the stamen (424), leaf (345), tepal (196), stigma (177), and corm (133) underscores the intricate regulatory networks governing source-to-sink partitioning and dynamic metabolic processes. Notably, our study identified several bioactive compounds, including crocin, picrocrocin, crocetin, safranal, cannabielsoin, quercetin, prenylnaringenin, isorhamnetin, pelargonidin, kaempferol, and gallic acid, all of which exhibit potential therapeutic properties. In conclusion, this comprehensive analysis significantly enhances our understanding of the molecular mechanisms driving the biosynthesis of apocarotenoids, cannabinoids, anthocyanins, and flavonoids in saffron, thereby providing valuable insights and paving the way for future research in this area., (© 2024 Scandinavian Plant Physiology Society.)

Background: Camellia nitidissima Chi is a popular ornamental plant because of its golden flowers, which contain flavonoids and carotenoids. To understand the regulatory mechanism of golden color formation, the metabolites of C. nitidissima petals at five different developmental stages were detected, a proteome map of petals was first constructed via tandem mass tag (TMT) analysis, and the accuracy of the sequencing data was validated via parallel reaction monitoring (PRM)., Results: Nineteen color components were detected, and most of these components were carotenoids that gradually accumulated, while some metabolites were flavonoids that were gradually depleted. A total of 97,647 spectra were obtained, and 6,789 quantifiable proteins were identified. Then, 1,319 differentially expressed proteins (DEPs) were found, 55 of which belong to the flavonoid and carotenoid pathways, as revealed by pairwise comparisons of protein expression levels across the five developmental stages. Notably, most DEPs involved in the synthesis of flavonoids, such as phenylalanine ammonium lyase and 4-coumarate-CoA ligase, were downregulated during petal development, whereas DEPs involved in carotenoid synthesis, such as phytoene synthase, 1-deoxy-D-xylulose-5-phosphate synthase, and β-cyclase, tended to be upregulated. Furthermore, protein‒protein interaction (PPI) network analysis revealed that these 55 DEPs formed two distinct PPI networks closely tied to the flavonoid and carotenoid synthesis pathways. Phytoene synthase and chalcone synthase exhibited extensive interactions with numerous other proteins and displayed high connectivity within the PPI networks, suggesting their pivotal biological functions in flavonoid and carotenoid biosynthesis., Conclusion: Proteomic data on the flavonoid and carotenoid biosynthesis pathways were obtained, and the regulatory roles of the DEPs were analyzed, which provided a theoretical basis for further understanding the golden color formation mechanism of C. nitidissima., (© 2024. The Author(s).)

Flavonoids, phenolic acids and terpenoids are important active ingredients that are biomarkers for evaluating the quality of Zizyphus jujuba Mill. Cv. Goutou jujube fruit. Nevertheless, regulatory network of these active ingredients biosynthesis in jujube fruit is still unclear. Here, integrated metabolomics and transcriptomics analyses were conducted at four different stages during the ripening of jujube fruits. Cytochrome P450 enzymes (CYP450s) and uridine 5'-diphospho-glucuronosyltransferases (UGTs) are pivotal enzymes for flavonoids, phenolic acids and terpenoids biosynthesis in plants. Benzoylmalic acid, a phenolic acid, 4', 5, 7-trihydroxyflavanon and quercetin-3-O-(6″-p-coumaroyl), two flavonoid metabolites, and jujuboside B1, a triterpenoid metabolite were targeted as they were correlated with both CYP450s and UGTs. Furthermore, networks of TFs, CYP450s and UGTs involved in the target metabolites biosynthesis were elucidated. NAC_1 and bZIP2 up-regulated CYP71A7 expression, while G2-like2 and bHLH_1 positively regulated the CSE expression contributing to promoted benzoylmalic acid biosynthesis. G2-like2, bHLH_1 and bHLH_2 indicated a positive relationship with CYP93D1, CYP86C2/3 or UGT71A16 which were positively correlated with 4', 5, 7-trihydroxyflavanon biosynthesis. MYB1/2/3, C2H2_2 and WRKY positively regulated expression of CYP82A4 or UGT_1 resulted in increased quercetin-3-O-(6″-p-coumaroyl) galactoside biosynthesis. G2-like2 and bHLH_1 up-regulated 4 C L, CYP93D1 or UGT71A16 was the reason for an increase of jujuboside B1 biosynthesis. The findings provide new insight into molecular breeding of high-quality jujube fruits., 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 Masson SAS.)

Aluminium (Al) stress is the second-leading abiotic stress on crops. An improved understanding of the response mechanisms of plants to Al stress will provide scientific guidance for enhancing the crops' tolerance to Al stress. In this study, Al stress (50-200 μM AlCl 3 ) caused visible damage to broad bean (Vicia faba L.) roots rather than shoots, which was attributed to Al accumulation and distribution in different tissues. Root transcriptomic analysis revealed that Al stress altered cell wall properties by downregulating lignin synthesis and several xyloglucan endotransglucosylase/hydrolase-, expansin- and peroxidase (POD)-encoding genes, which likely weakened cell extensibility to inhibit root growth. Additionally, Al stress impeded reactive oxygen species scavenging pathways involving POD activity and flavonoid biosynthesis, leading to oxidative damage characterised by malondialdehyde accumulation. These results indicate that optimising cell wall properties and/or enhancing antioxidant processes are crucial for alleviating Al toxicity to broad beans. Interestingly, exogenous application (500 and 1000 μM) of the flavonoid apigenin effectively alleviated Al toxicity in broad bean roots by partially improving the total antioxidant capacity of the roots. This study contributes to understanding the interaction between plants and Al and provides new strategies to alleviate Al toxicity in crops., 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 Masson SAS. All rights reserved.)

Plant adaptation from aquatic to terrestrial environments required modifications to cell wall structure and function to provide tolerance to new abiotic and biotic stressors. Here, we investigate the nature and function of red auronidin pigment accumulation in the cell wall of the liverwort Marchantia polymorpha. Transgenic plants with auronidin production either constitutive or absent were analysed for their cell wall properties, including fractionation of polysaccharide and phenolic components. While small amounts of auronidin and other flavonoids were loosely associated with the cell wall, the majority of the pigments were tightly associated, similar to what is observed in angiosperms for polyphenolics such as lignin. No evidence of covalent binding to a polysaccharide component was found: we propose auronidin is present in the wall as a physically entrapped large molecular weight polymer. The results suggested auronidin is a dual function molecule that can both screen excess light and increase wall strength, hydrophobicity and resistance to enzymatic degradation by pathogens. Thus, liverworts have expanded the core phenylpropanoid toolkit that was present in the ancestor of all land plants, to deliver a lineage-specific solution to some of the environmental stresses faced from a terrestrial lifestyle., (© 2024 The Author(s). The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

The diversity of secondary metabolites is an important means for plants to cope with the complex and ever-changing terrestrial environment. Plant biosynthetic gene clusters (BGCs) are crucial for the biosynthesis of secondary metabolites. The domestication and evolution of BGCs and how they affect plant secondary metabolites biosynthesis and environmental adaptation are still not fully understood. Buckwheat exhibits strong resistance and abundant secondary metabolites, especially flavonoids, allowing it to thrive in harsh environments. A non-canonical BGC named UFGT3 cluster is identified, which comprises a phosphorylase kinase (PAK), two transcription factors (MADS1/2), and a glycosyltransferase (UFGT3), forming a complete molecular regulatory module involved in flavonoid biosynthesis. This cluster is selected during Tartary buckwheat domestication and is widely present in species of the Fagopyrum genus. In wild relatives of cultivated buckwheat, a gene encoding anthocyanin glycosyltransferase (AGT), which glycosylates pelargonidin into pelargonidin-3-O-glucoside, is found inserted into this cluster. The pelargonidin-3-O-glucoside can help plants resist UV stress, endowing wild relatives with stronger high-altitude adaptability. This study provides a new research paradigm for the evolutionary dynamics of plant BGCs, and offers new perspectives for exploring the mechanism of plant ecological adaptability driven by environmental stress through the synthesis of secondary metabolites., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

UDP-glycosyltransferases (UGTs) play a crucial role in the glycosylation of secondary metabolites in plants, which is of significant importance for growth and response to biotic or abiotic stress. Despite the wide identification of UGT family members in various species, limited information is available regarding this family in citrus. In this study, we identified 87 UGT genes from the Citrus sinensis genome and classified them into 14 groups. We characterized their gene structures and motif compositions, providing insights into the molecular basis underlying discrepant functions of UGT genes within each evolutionary branch. Tandem duplication events were found to be the main driving force behind UGT gene expansion. Additionally, we identified numerous cis-acting elements in the promoter region of UGT genes, including those responsive to light, growth factors, phytohormones, and stress conditions. Notably, light-responsive elements were found with a frequency of 100 %. We elucidated the expression pattern of UGTs during fruit development in Citrus aurantium using RNA-seq and quantitative real-time PCR (qRT-PCR), revealing that 10 key UGT genes are closely associated with biosynthesis of bitter flavanone neohesperidosides (FNHs). Furthermore, we identified Ca1,2RhaT as a flavonoid 1-2 rhamnosyltransferase (1,2RhaT) involved in FNHs biosynthesis for the first time. Isolation and functional characterization of the gene Ca1,2RhaT from Citrus aurantium in vitro and in vivo indicated that Ca1,2RhaT encoded a citrus 1,2RhaT and possessed rhamnosyl transfer activities. This work provides comprehensive information on the UGT family while offering new insights into understanding molecular mechanisms regulating specific accumulation patterns of FNHs or non-bitter flavanone rutinosides (FRTs) in citrus., 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.)

Mung beans were pretreated with a combination of ultrasonic and calcium ion to enhance the polyphenol content and antioxidant capacity during germination. Changes in polyphenol content and antioxidant capacity during germination, along with underlying mechanisms, were investigated. Both single ultrasound and combined ultrasound-Ca 2+ pretreatments significantly increased the polyphenol content and enhanced the antioxidant capacity (p < 0.05) of mung beans depending on germination period. Among 74 polyphenolic metabolites identified in germinated mung beans, 50 were differential. Notably, 23 of these metabolites showed a significant positive correlation with antioxidant activity. Ultrasound pretreatment promoted flavonoid biosynthesis, whereas ultrasound-Ca 2+ pretreatment favored the tyrosine synthesis pathway. Polyphenol composition and accumulation changes were mainly influenced by metabolic pathways like flavonoid, isoflavonoid, anthocyanin, and flavone/flavonol biosynthesis. The results suggest that ultrasound alone or combined with calcium ion pretreatments effectively enhance mung bean polyphenol content and antioxidant capacity during germination., 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 Ltd. All rights reserved.)

The phenylpropanoid pathway is vital for plant growth and development, producing lignin and flavonoids. This study investigates PtrMYB203, a homolog of MYB repressors of proanthocyanidin (PA) biosynthesis in Populus trichocarpa, as a transcriptional repressor in the phenylpropanoid pathway of hybrid poplar (Populus alba x P. glandulosa). Overexpression of PtrMYB203 (35S::PtrMYB203) in hybrid poplar detrimentally impacted plant growth and development. Histological analysis revealed irregular xylem vessel formation and decreased lignin content, corroborated by Klason lignin assays. Moreover, 35S::PtrMYB203 transgenic poplars exhibited significant decreases in anthocyanin and PA accumulations in callus tissues, even under high light conditions. Quantitative RT-PCR analysis and protoplast-based transcriptional activation assay confirmed the downregulation of lignin and flavonoid biosynthesis genes. This genetic modification also alters the expression of several MYB transcription factors, essential for phenylpropanoid pathway regulation. Remarkably, saccharification efficiency in the 35S::PtrMYB203 poplar was improved by over 34% following hot water treatment alone. These findings suggest PtrMYB203 as a potential genetic target for enhancing wood properties for bioenergy production, providing valuable insights into the manipulation of metabolite pathways in woody perennials to advance wood biotechnology., 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 Masson SAS. All rights reserved.)

Plant responses to cold stress include either induction of flavonoid biosynthesis as part of defense responses or initially elevated levels of these substances to mitigate sudden temperature fluctuations. The role of chromatin modifying factors and, in general, epigenetic variability in these processes is not entirely clear. In this work, we review the literature to establish the relationship between flavonoids, cold and chromatin modifications. We demonstrate the relationship between cold acclimation and flavonoid accumulation, and then describe the cold adaptation signaling pathways and their relationship with chromatin modifying factors. Particular attention was paid to the cold signaling module OST1-HOS1-ICE1 and the novel function of the E3 ubiquitin protein ligase HOS1 (a protein involved in chromatin modification during cold stress) in flavonoid regulation., 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 Masson SAS. All rights reserved.)

Lilium spp. is a world-famous bulbous flower with outstanding ornamental, edible, and medicinal value. Evaluating the taste of edible lilies and identifying important active substances and genes are necessary for germplasm improvement, new variety breeding, and industrial application. To better understand the phenylpropanoids and regalosides biosynthesis, L. davidii var. unicolor and L. lancifolium Thunb. bulbs were used for transcriptome and metabolite analysis. Results showed that the phenols and flavonoid contents in JT were lower than in LT, while the saponins and alkaloid contents in JT were higher than in LT. A total of 20,520 differentially expressed genes (DEGs) and 383 differential metabolites were searched. Integrated transcriptomics and metabolomics analysis showed that phenylpropanoid biosynthesis and flavonoid biosynthesis were differentially altered. Ninety-nine unigenes encoding ten phenolic acids and two flavonoids were identified as candidate genes involved in phenylpropanoid and flavonoid biosynthesis. WGCNA analysis showed 76 phenylpropanoid and flavonoid biosynthesis-related unigenes were verified as likely to be involved in phenylpropanoid metabolism and regalosides accumulation. Among them, 15 genes were used for qRT-PCR, and four genes were utilized for tissue-specific expression pattern analysis. Down-regulation of LdPAL2 and LdC4H1 in bulbs of L. davidii var. unicolor via virus induced gene silence (VIGS) reduced the contents of p-coumaric acid and cinnamic acid. These results contribute to understanding phenylpropanoid metabolism and identifying potential functional genes for improving the regalosides and flavonoids content in Lilium bulbs., Competing Interests: Declaration of competing interest The authors declare that there are no potential conflicts or competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Banisteriopsis (Malpighiaceae) is an important genus of neotropical savannas with related biological and medicinal activities but under-explored metabolomic profiles. We present a chemometric analysis for discriminating secondary metabolites of three species of Banisteriopsis (B. laevifolia, B. malifolia, and B. stellaris) leaves. Initially, each species was separately extracted with ethanol:water (4 : 1, v/v) and analysed by Ultra Performance Liquid Chromatography coupled with Mass Spectrometry (UPLC-MS/MS). The chromatographic profiles were subjected to Global Natural Product Social (GNPS) and Partial Least Squares Discriminant Analysis (PLS-DA). Eighty-nine compounds (cosine≥0.90) were annotated, including flavonoids, phenolics, and acids. The chemometric analysis (VIP Score) showed each species' relative concentration of the more relevant compounds. In addition, four compounds that discriminate the metabolomic profiles of B. laevifolia, B. malifolia, and B. stellaris were identified by PLS-DA., (© 2024 Crown copyright and The Author(s). Chemistry & Biodiversity published by Wiley-VHCA AG. This article is published with the permission of the Controller of HMSO and the King's Printer for Scotland.)

Flavonoid compounds are widely present in various organs and tissues of different plants, playing important roles when plants are exposed to abiotic stresses. Different types of flavonoids are biosynthesized by a series of enzymes that are encoded by a range of gene families. In this study, a total of 63 flavonoid pathway genes were identified from the genome of Medicago truncatula. Gene structure analysis revealed that they all have different gene structure, with most CHS genes containing only one intron. Additionally, analysis of promoter sequences revealed that many cis-acting elements responsive to abiotic stress are located in the promoter region of flavonoid pathway genes. Furthermore, analysis on M. truncatula gene chip data revealed significant changes in expression level of most flavonoid pathway genes under the induction of salt or drought treatment. qRT-PCR further confirmed significant increase in expression level of several flavonoid pathway genes under NaCl and mannitol treatments, with CHS1, CHS9, CHS10, F3'H4 and F3'H5 genes showing significant up-regulation, indicating they are key genes in response to abiotic stress in M. truncatula. In summary, our study identified key flavonoid pathway genes that were involved in salt and drought response, which provides important insights into possible modification of flavonoid pathway genes for molecular breeding of forage grass with improved abiotic resistance., 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 GmbH. All rights reserved.)

Tilianin and linarin, two rare glycosylated flavonoids in the aromatic endangered medicinal plant Nardostachys jatamansi (D.on)DC., play an important role in the fields of medicine, cosmetics, food and dye industries. However, there remains a lack of comprehensive understanding regarding their biosynthetic pathway. In this study, the phytochemical investigation of N. jatamansi resulted in the isolation of linarin. With help of AlphaFold2 to cluster the entire glycosyltransferase family based on predicted structure similarities, we successfully identified a flavonoid glycosyltransferase NjUGT73B1, which could efficiently catalyze the glucosylation of acacetin at 7-OH to produce tilianin, also the key precursor in the biosynthesis of linarin. Additionally, NjUGT73B1 displayed a high degree of substrate promiscuity, enabling glucosylation at 7-OH of many flavonoids. Molecular modeling and site-directed mutagenesis revealed that H19, H21, H370, F126, and F127 play the crucial roles in the glycosylation ability of NjUGT73B1. Notably, comparation with the wild NjUGT73B1, mutant H19K led to a 50% increase in the activity of producing tilianin from acacetin., 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 Ltd. All rights reserved.)

The identification of new genes involved in regulating cold tolerance in rice is urgent because low temperatures repress plant growth and reduce yields. Cold tolerance is controlled by multiple loci and involves a complex regulatory network. Here, we show that rice jacalin-related lectin (OsJRL) modulates cold tolerance in rice. The loss of OsJRL gene functions increased phenylalanine metabolism and flavonoid biosynthesis under cold stress. The OsJRL knock-out (KO) lines had higher phenylalanine ammonia-lyase (PAL) activity and greater flavonoid accumulation than the wild-type rice, Nipponbare (NIP), under cold stress. The leaves had lower levels of reactive oxygen species (ROS) and showed significantly enhanced cold tolerance compared to NIP. In contrast, the OsJRL overexpression (OE) lines had higher levels of ROS accumulation and showed lower cold tolerance than NIP. Additionally, the OsJRL KO lines accumulated more abscisic acid (ABA) and jasmonic acid (JA) under cold stress than NIP. The OsJRL OE lines showed increased sensitivity to ABA compared to NIP. We conclude that OsJRL negatively regulates the cold tolerance of rice via modulation of phenylalanine metabolism and flavonoid biosynthesis., (© 2024 John Wiley & Sons Ltd.)

Lactobacillus strains have emerged as promising probiotics for enhancing the bioactivities of plant-based foods associated with flavonoid biotransformation. Employing microbial fermentation and mass spectrometry, we explored flavonoid metabolism in lychee pulp fermented separately by Lactiplantibacillus plantarum and Limosilactobacillus fermentum. Two novel metabolites, 3,5,7-trihydroxychromone and catechol, were exclusively identified in L. plantarum-fermented pulp. Concomitant with consumption of catechin and quercetin glycosides, dihydroquercetin glycosides, 2,4-dihydroxybenzoic acid and p-hydroxyphenyllactic acid were synthesized by two strains through hydrogenation and fission of C-ring. Quantitative analysis revealed that bound phenolics were primarily located in water-insoluble polysaccharides in lychee pulp. Quercetin 3-O-rutinoside was partially liberated from water-insoluble polysaccharides and migrated to water-soluble polysaccharides during fermentation. Meanwhile, substantial accumulations in short-chain fatty acids (increased 1.45 to 3.08-fold) and viable strains (increased by 1.97 to 2.00 Log 10 CFU/mL) were observed in fermentative pulp. These findings provide broader insight into microbial biotransformation of phenolics and possible guidance for personalized nutrition., 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 Ltd. All rights reserved.)

Blueberries are vulnerable to chilling injury (CI). This can lead to limited longevity when they are subjected to cold storage conditions. This study investigated the effectiveness of a preharvest spray containing 0.02% hexanal in reducing CI and improving the postharvest storage quality of 'Star' and 'Biloxi' blueberries. The blueberries were stored for a period of 5 weeks at 2 °C and in 90% relative humidity (RH). The findings revealed that the preharvest hexanal spraying of both cultivars delayed senescence by mitigating CI, as evidenced by the bolstering of the antioxidant defense system through increased superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), catalase (CAT), and phenylalanine ammonia lyase (PAL) enzyme activity. The treated fruit also maintained elevated levels of total phenol content (TPC), total flavonoids (TFC), and vitamin C, demonstrating enhanced free radical scavenging capacity (FRSC), while exhibiting reduced polyphenoloxidase (PPO) activity, and reduced malondialdehyde (MDA), and H 2 O 2 content in comparison with the control group. The preharvest hexanal treatment also suppressed fruit softening by maintaining greater firmness and higher membrane stability index (MSI) scores, inhibiting the activity of polygalacturonase (PG), pectinmethylesterase (PME), xylanase, and α-amylase, and reducing microbial counts (MC) and incidence of decay (DI) in comparison with the control. Preharvest hexanal treatment also improved the overall storage quality by reducing weight loss, total soluble solids (TSS), pH, and the TSS/acid ratio, while increasing titratable acidity (TA) in comparison with the control during cold storage. The findings suggest that hexanal, as a preharvest application, delays senescence effectively and preserves overall quality by enhancing cold tolerance through antioxidant defense mechanisms in blueberry storage under cold conditions. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Salvia miltiorrhiza and Salvia prattii seeds are rich in metabolites that are beneficial to human health and can be utilised as nutritional supplements. In this study, UPLC-MS and GC-MS based on extensively focused metabolomics were used to compare the seed metabolomics of the two species. LC-MS detected 118 metabolites, primarily Lipids and phenylpropanoids. GC- MS detected a total of 188 metabolites, mainly organic acids and their derivatives, of which Salvia prattii seeds contain high levels of nutrients. In addition, we experimentally determined antioxidant activity of two Salvia species, and the results showed that the antioxidant activity of Salvia prattii seeds was about twice as high as that of Salvia miltiorrhiza seeds. We used WGCNA to group the metabolites, and found the central metabolites in the focal modules including flavonoids and terpenoids. Our study contributes valuable knowledge for future research on the chemical makeup of Salvia prattii seeds.

Round green tea (RGT) presents unique properties and is widely distributed in China, and during processing, it undergoes dynamic changes in non-volatile metabolites (NVMs), which are poorly understood. Utilizing UHPLC-Q-Exactive/MS analysis, this study comprehensively characterized 216 NVMs during RGT processing and identified fixation and pan-frying as key processes influencing NVMs. Additionally, 23 key differential NVMs were screened, with amino acid and flavonoid metabolism highlighted as key metabolic pathways for RGT taste and color quality. The impact of pan-frying degree on shape, color, and taste was also explored. Moderate pan-frying led to optimal results, including a tight and round shape, green and bright color, mellow and umami taste, and reduced astringent and bitter taste NVMs, including epigallocatechin gallate, procyanidin B2, myricetin 3-O-galactoside, quinic acid, strictinin, phenylalanine, and theobromine. This study addresses the NVM research gap in RGT processing, thus providing a technical foundation for the precision-oriented processing of high-quality tea., Competing Interests: Declaration of competing interest The authors declare no potential conflicts of interest regarding the research, authorship, and/or publication of this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Rice (Oryza sativa L.) husk harbors a substantial proportion of biological metabolites, as one of the most plentiful agriculture by-products in rice milling process, rice husk remains poorly utilized. As a continuing search for potential bioactive molecules from the husk of rice, a totally of twelve components (1-12), including six sterol ferulates (1-6), one flavonoid (7), one dipeptide (8), and four phenylpropanoid derivatives (9-12) were obtained. All the chemical structures were elucidated based on comprehensive spectroscopic data. Wherein, compounds 1 and 2 were yield as previous undescribed metabolites, and the comprehensive NMR data for compounds 3 and 4 were first presented in its entirety. Motivated by the similarity of the structural motifs of components 1-6 to that of reported sterol ferulates, the antioxidant and anti-inflammatory effects for compounds 1-6 were evaluated in vitro. Among them, compounds 5/6 had a significant antioxidant activity compare to that of vitamin E in both DPPH and reducing power assay up to the concentration 40 μg/ml; while compounds 1 and 2 exhibited weak suppressive effect on the production of nitric oxide, with the IC 50 values of 53.27±1.37 μM., (© 2024 Wiley-VHCA AG, Zurich, Switzerland.)

Background: Kratom (Mitragyna speciosa) has a long history of traditional use. It contains various alkaloids and polyphenols. The properties of kratom's alkaloids have been well-documented. However, the property of kratom's polyphenols in water-soluble phase have been less frequently reported. This study assessed the effects of water-soluble Mitragyna speciosa (kratom) extract (MSE) on gut microbiota and their metabolite production in fecal batch culture., Results: The water-soluble kratom extract (MSE0) and the water-soluble kratom extract after partial sugar removal (MSE50) both contained polyphenols, with total phenolic levels of 2037.91 ± 51.13 and 3997.95 ± 27.90 mg GAE/g extract, respectively and total flavonoids of 81.10 ± 1.00 and 84.60 ± 1.43 mg CEQ/g extract. The gut microbiota in fecal batch culture was identified by 16S rRNA gene sequencing at 0 and 24 h of fermentation. After fermentation, MSE50 stimulated the growth of Bifidobacterium more than MSE0. MSE0 gave the highest total fatty acids level among the treatments. The phenolic metabolites produced by some intestinal microbiota during fecal fermentation at 24 h were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The major metabolite of biotransformation of both water-soluble MSEs by intestinal microbiota was pyrocatechol (9.85-11.53%)., Conclusion: The water-soluble MSEs and their produced metabolites could potentially be used as ingredients for functional and medicinal food production that supports specific gut microbiota. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Slow-release urea, as an environmentally friendly fertiliser, can provide a continuous and uniform supply of nutrients needed by the crop, reduce the amount and frequency of fertiliser application, and promote the uptake and utilisation of nitrogen in crops. The production of E. ferox is often dominated by the application of quick-acting fertilisers, resulting in serious problems of over-fertilisation, inappropriate periods of fertilisation, eutrophication of soil and water due to fertilisation, and difficulties in applying fertilisers. Therefore, in this study, different amounts (CK, T1, T2, T3, T4, T5) of SRU (Slow-release Urea) were first applied, and T3 (18.8 kg·667 m -2 ) was found to significantly improve both yield and quality. Further, it was found that under different SRU (CK, S1, S2, S3, S4) application period treatments, application of 18.8 kg·667 m -2 at AFP20 (S2) period significantly increased the yield and quality of E. ferox . In the seed kernels of E. ferox , the total yield, soluble sugar content, total starch, and flavonoid content increased significantly by 10.35%, 36.40%, 5.91%, and 22.80%, respectively, compared with CK. In addition, the expression of key sugar transporter genes ( EfSWEETs ), flavonoid synthesis-related genes ( EfPAL , EfDFR , etc.), and starch synthesis-related enzyme activities (SBE, SSS, GBSS) were significantly increased. By exploring the quantity of application and application period of SRU, this study was carried out to investigate the in-depth effect of SRU on the growth and development of E. ferox and to provide technical references for the increase in E. ferox yield, the improvement in E. ferox quality, and the simplification of fertiliser application.

Pulsed light (PL) is a prospective non-thermal technology that can improve the degradation of ginkgolic acid (GA) and retain the main bioactive compounds in Ginkgo biloba leaves (GBL). However, only using PL hasn't yet achieved the ideal effect of reducing GA. Fermentation of GBL to make ginkgo dark tea (GDT) could decrease GA. Because different microbial strains are used for fermentation, their metabolites and product quality might differ. However, there is no research on the combinative effect of PL irradiation fixation and microbial strain fermentation on main bioactive compounds and sensory assessment of GDT. In this research, first, Bacillus subtilis and Saccharomyces cerevisiae were selected as fermentation strains that can reduce GA from the five microbial strains. Next, the fresh GBL was irradiated by PL for 200 s (fluences of 0.52 J/cm 2 ), followed by B. subtilis, S. cerevisiae, or natural fermentation to make GDT. The results showed that compared with the control (unirradiated and unfermented GBL) and the only PL irradiated GBL, the GA in GDT using PL + B. subtilis fermentation was the lowest, decreasing by 29.74%; PL + natural fermentation reduced by 24.53%. The total flavonoid content increased by 14.64% in GDT using PL + B. subtilis fermentation, whose phenolic and antioxidant levels also increased significantly. Sensory evaluation showed that the color, aroma, and taste of the tea infusion of PL + B. subtilis fermentation had the highest scores. In conclusion, the combined PL irradiation and solid-state fermentation using B. subtilis can effectively reduce GA and increase the main bioactive compounds, thus providing a new technological approach for GDT with lower GA., 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 © 2023. Published by Elsevier Ltd.)

Neglected and underutilised plants such as Pseudocydonia sinensis (Chinese quince) have garnered global interest as invaluable sources of natural bioactive compounds. Herein, a wide-targeted metabolomics-based approach revealed 1199 concurrent metabolites, with further analysis of their fluctuations across with the five stages of fruit growth. The bioactive compounds in Chinese quince primarily comprised sugars and organic acids, flavonoids, and terpenoids. Moreover, 395 metabolites were identified as having medicinal properties and rutin was the most content of them. Transcriptome analysis further provided a molecular basis for the metabolic changes observed during fruit development. By thoroughly analysing metabolite and transcriptome data, we revealed changes in bioactive compounds and related genes throughout fruit development. This study has yielded valuable insights into the ripening process of Chinese quince fruit, presenting substantial implications for industrial applications, particularly in quality control., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Despite being a highly nutritious and resilient cereal, pearl millet is not popular among consumers and food industries due to the short shelf-life of flour attributed to rapid rancidity development. The biochemical mechanism underlying rancidity, a complex and quantitative trait, needs to be better understood. The present study aims to elucidate the differential accumulation of metabolites in pearl millet that impact the rancidity process. Metabolite profiling was conducted on ten pearl millet genotypes with varying levels of rancidity-comprising high, low, and medium rancid genotypes-utilizing liquid chromatography and high-resolution mass spectrometry (LC-HRMS) at different accelerated ageing conditions. Through non-targeted metabolomic analysis, crucial metabolites associated with rancidity were identified across various biochemical pathways, including fatty acids, glycerophospholipids, sphingolipids, glycerol lipids, flavonoids, alkaloids, and terpenoids. Notably, metabolites such as fatty aldehydes, fatty alcohols, fatty esters, fatty acyls, fatty esters, and fatty amides were significantly elevated in high rancid genotypes, indicating their involvement in the rancidity process. These fatty acids-related metabolites further break down into saturated and unsaturated fatty acids. Four key fatty acids-stearic, palmitic, linoleic and linolenic acid-were quantified in the ten pearl millet genotypes, confirming their role in rancidity development. This investigation promises novel insights into utilizing metabolomics to understand the biochemical processes and facilitate precision breeding for developing low-rancidity pearl millet lines.

This study investigated postharvest dopamine treatment efficiency in ameliorating chilling injury of banana fruits during storage at 7 ºC for 21 days. Our results showed that dopamine treatment at 150 µM promoted phenols and flavonoids biosynthesis acquired by higher phenylalanine ammonia-lyase (PAL) expression and activity concurrent with lower polyphenol oxidase (PPO) expression and activity leading to higher DPPH, FRAP, and ABTS radicals scavenging activity. In addition, dopamine treatment at 150 µM promoted endogenous proline biosynthesis by activating pyrroline-5-carboxylate synthetase (P5CS) and ornithine δ-aminotransferase (OAT) expression and activity concurrent with suppressing proline dehydrogenase (ProDH) expression and activity. Furthermore, higher endogenous γ-aminobutyric acid (GABA) biosynthesis in banana fruits by 150 µM dopamine treatment was accompanied by higher glutamate decarboxylase (GAD) and GABA transaminase (GABA-T) expression and activity. Therefore, our results suggest that dopamine treatment at 150 µM might be employed for banana fruits chilling injury amelioration by enhancing phenylpropanoid pathway activity and boosting endogenous proline and GABA biosynthesis., (© 2024. The Author(s).)

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.

The complex of oat β-glucan (OBG) and flavonoids hampered the digestion of starch-based food and retarded the blood glucose response; however, its effect on gastric emptying and its relative mechanism have not been thoroughly investigated. By using Fourier transform infrared (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), antioxidant ability, and enzymic inhibitory tests for the characterization and in vitro semi-dynamic digestion of complexes of OBG (high and low molecular weights) and sea buckthorn flavonoids, we found that the higher molecular weight complex (FU) exhibited stronger ABTS and DPPH radical scavenging abilities and higher α-glucosidase and α-amylase inhibition rates. Mice fed with rice flour with FU addition exhibited the slowest gastric emptying and intestinal propulsion rates and blood glucose rise and had the lowest activity of digestive enzymes and levels of insulin, ghrelin, motilin (MTL), and relevant gene (ghrelin and GHSR mRNA) expression than those in the control and low-molecular-weight groups. This study provided scientific data for the development of foods with delayed gastric rate and hypoglycemic index for specific populations.

Duckweeds (Spirodela polyrrhiza) are free-floating macrophytes that grow profusely in nutrient-rich waters. Under ideal conditions, they exhibit a rapid growth rate and can absorb a substantial amount of nutrients, macromolecules, and pollutants from bodies of water. Zinc oxide nanoparticles (ZnO NPs) synthesized from plant extracts, particularly under stress conditions, have opened new research avenues in the field of nanotechnology. Under salinity stress, the accumulation of flavonoids in duckweeds can affect the structure of ZnO NPs, helping researchers ascertain their antimicrobial role. In our study, we exposed mid-log phase duckweed monocultures to 75 mM NaCl in a full-strength Murashige and Skoog medium for 7 days, followed by a 15-day recovery period. We observed significant overexpression of superoxide and hydrogen peroxide as reactive oxygen species. As a result, chlorophyll and certain metabolites were produced in lesser amounts, while flavonoid and phenol content increased by 12% and 8%, respectively. This overproduction persisted up to 10 days into the recovery treatment period but dropped by 8% and 5%, respectively, by the 15th day. The flavonoid coating transformed the NPs into rosette clusters, which exhibited reduced antimicrobial activity against Aeromonas hydrophila, a Gram-negative, fish-pathogenic bacterium. Herein, we discuss potential mechanisms for the conformational transformation of ZnO NPs into finer dimensions in response to NaCl-induced oxidative stress in duckweed. In this study, the azo dye degradation capacity of salinity-treated plants increased as the flavonoid profile became enriched. Zinc oxide nanoparticles, both prior to and after salinity treatment, were found to be efficient in scavenging azo dye and mitigating its toxicity, as evidenced by improved germination, growth, and overall plant morphometry., (© 2024. The Author(s).)

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: The utilization of nutrition management, has recently been developed as a means of improving the growth and production of phytochemical compounds in herbs. The present study aimed to improve the growth, physiological, and phytochemical characteristics of lemon balm (Melissa officinalis L.) using different NH 4 + (ammonium) to NO 3 ̄ (nitrate) ratios (0:100, 25:75, 50:50, 75:25 and 100:0) under floating culture system (FCS)., Results: The treatment containing 0:100 - NH 4 + :NO 3 ̄ ratio showed the most remarkable values for the growth and morpho-physiological characteristics of M. officinalis. The results demonstrated that maximum biomass (105.57 g) earned by using the ratio of 0:100 and minimum at 75:25 ratio of NH 4 + : NO 3 ̄. The plants treated with high nitrate ratio (0:100 - NH 4 + :NO 3 ̄) showed the greatest concentration of total phenolics (60.40 mg GAE/g DW), chlorophyll a (31.32 mg/100 g DW), flavonoids (12.97 mg QUE/g DW), and carotenoids (83.06 mg/100 g DW). Using the 75:25 - NH 4 + :NO 3 ̄ ratio caused the highest dry matter (DM), N and K macronutrients in the leaves. The highest antioxidant activity by both DPPH (37.39 µg AAE/mL) and FRAP (69.55 mM Fe ++ /g DW) methods was obtained in 75:25 - NH 4 + :NO 3 ̄ treatment. The p-coumaric acid as a main abundant phenolic composition, was detected by HPLC analysis as the highest content in samples grown under 0:100 - NH 4 + :NO 3 ̄ treatment. Also, the major compounds in M. officinalis essential oil were identified as geranial, neral, geranyl acetate and geraniol by GC analysis. With increasing NO 3 ̄ application, geraniol and geranyl acetate contents were decreased., Conclusions: The findings of present study suggest that the management of NH4 + to NO 3 ̄ ratios in nutrient solutions could contribute to improving growth, physiological and phytochemical properties of M. officinalis. The plants treated with high nitrate ratio (especially 0:100 - NH 4 + :NO 3 ̄) showed the greatest effects on improving the growth and production of morpho-physiological and phytochemical compounds. By comprehensively understanding the intricate dynamics among nitrogen sources, plants, and their surroundings, researchers and practitioners can devise inventive approaches to optimize nitrogen management practices and foster sustainable agricultural frameworks., (© 2024. The Author(s).)

Background: Glechoma longituba, recognized as a medicinal plant, provides valuable pharmaceutical raw materials for treating various diseases. Saline-alkali stress may effectively enhance the medicinal quality of G. longituba by promoting the synthesis of secondary metabolites. To investigate the changes in the primary medicinal components of G. longituba under saline-alkali stress and improve the quality of medicinal materials, Na 2 CO 3 was applied to induce short-term stress under different conditions and the biomass, physiologically active substances and primary medicinal components of G. longituba were measured in this study., Results: Under alkaline salt stress, the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) were elevated in G. longituba, accompanied by increased accumulation of proline (Pro) and malondialdehyde (MDA). Furthermore, analysis of the medicinal constituents revealed that G. longituba produced the highest levels of soluble sugars, flavonoids, ursolic acid, and oleanolic acid under 0.6% Na 2 CO 3 stress for 48 h, 0.2% Na 2 CO 3 stress for 72 h, 0.4% Na 2 CO 3 stress for 12 h, and 0.4% Na 2 CO 3 stress for 8 h, respectively., Conclusions: Short-term Na 2 CO 3 stress enhances the synthesis of medicinal components in G. longituba. By manipulating stress conditions, the production of various medicinal substances could be optimized. This approach may serve as a basis for the targeted cultivation of G. longituba, offering potential applications in the treatment of diverse diseases., (© 2024. The Author(s).)

Eurotium cristatum, a unique probiotic in Fu brick tea, is widely used in food processing to enhance added values. Here, green kernel black beans (GKBBs) were solid-fermented with E. cristatum and dynamic changes in flavour, chemical composition and metabolites during fermentation were investigated. As results, E. cristatum fermentation altered aroma profiles and sensory attributes of GKBBs, especially reduced sourness. After fermentation, total polyphenolic and flavonoid contents in GKBBs were elevated, while polysaccharides, soluble proteins and short-chain fatty acids contents were decreased. E. cristatum fermentation also induced biotransformation of glycosidic isoflavones into sapogenic isoflavones. During fermentation, dynamic changes in levels of 17 amino acids were observed, in which 3 branched-chain amino acids were increased. Non-targeted metabolomics identified 51 differential compounds and 10 related metabolic pathways involved in E. cristatum fermentation of GKBBs. This study lays foundation for the development of green kernel black bean-based functional food products with E. cristatum fermentation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)