Flavonoid Biosynthesis Genes Research Articles

Metabolomic and Transcriptomic Analyses of Flavonoid Biosynthesis in Different Colors of Soybean Seed Coats.

Soybean has outstanding nutritional and medicinal value because of its abundant protein, oil, and flavonoid contents. This crop has rich seed coat colors, such as yellow, green, black, brown, and red, as well as bicolor variants. However, there are limited reports on the synthesis of flavonoids in the soybean seed coats of different colors. Thus, the seed coat metabolomes and transcriptomes of five soybean germplasms with yellow (S141), red (S26), brown (S62), green (S100), and black (S124) seed coats were measured. In this study, 1645 metabolites were detected in the soybean seed coat, including 426 flavonoid compounds. The flavonoids differed among the different-colored seed coats of soybean germplasms, and flavonoids were distributed in all varieties. Procyanidins A1, B1, B6, C1, and B2, cyanidin 3-O-(6″-malonyl-arabinoside), petunidin 3-(6″-p-coumaryl-glucoside) 5-glucoside, and malvidin 3-laminaribioside were significantly upregulated in S26_vs._S141, S62_vs._S141, S100_vs._S141, and S124_vs._S141 groups, with a variation of 1.43-2.97 × 1013 in terms of fold. The differences in the contents of cyanidin 3-O-(6″-malonyl-arabinoside) and proanthocyanidin A1 relate to the seed coat color differences of red soybean. Malvidin 3-laminaribioside, petunidin 3-(6″-p-coumaryl-glucoside) 5-glucoside, cyanidin 3-O-(6″-malonyl-arabinoside), and proanthocyanidin A1 affect the color of black soybean. The difference in the contents of procyanidin B1 and malvidin 3-glucoside-4-vinylphenol might be related to the seed coat color differences of brown soybeans. Cyanidin 3-gentiobioside affects the color of green soybean. The metabolomic-transcriptomic combined analysis showed that flavonoid biosynthesis is the key synthesis pathway for soybean seed color formation. Transcriptome analysis revealed that the upregulation of most flavonoid biosynthesis genes was observed in all groups, except for S62_vs._S141, and promoted flavonoid accumulation. Furthermore, CHS, CHI, DFR, FG3, ANR, FLS, LAR, and UGT88F4 exhibited differential expression in all groups. This study broadens our understanding of the metabolic and transcriptomic changes in soybean seed coats of different colors and provides new insights into developing bioactive substances from soybean seed coats.

Molecular and Metabolic Regulation of Flavonoid Biosynthesis in Two Varieties of Dendrobium devonianum.

Dendrobium devonianum is an important medicinal plant, rich in flavonoid, with various pharmacological activities such as stomachic and antioxidant properties. In this study, we integrated metabolome and transcriptome analyses to reveal metabolite and gene expression profiles of D. devonianum, both green (GDd) and purple-red (RDd) of semi-annual and annual stems. A total of 244 flavonoid metabolites, mainly flavones and flavonols, were identified and annotated. Cyanidin and delphinidin were the major anthocyanidins, with cyanidin-3-O-(6″-O-p-Coumaroyl) glucoside and delphinidin-3-O-(6″-O-p-coumaroyl) glucoside being the highest relative content in the RDd. Differential metabolites were significantly enriched, mainly in flavonoid biosynthesis, anthocyanin biosynthesis, and flavone and flavonol biosynthesis pathways. Transcriptomic analysis revealed that high expression levels of structural genes for flavonoid and anthocyanin biosynthesis were the main reasons for color changes in D. devonianum stems. Based on correlation analysis and weighted gene co-expression network analysis (WGCNA) analysis, CHS2 (chalcone synthase) and UGT77B2 (anthocyanidin 3-O-glucosyltransferase) were identified as important candidate genes involved in stem pigmentation. In addition, key transcription factors (TFs), including three bHLH (bHLH3, bHLH4, bHLH5) and two MYB (MYB1, MYB2), which may be involved in the regulation of flavonoid biosynthesis, were identified. This study provides new perspectives on D. devonianum efficacy components and the Dendrobium flavonoids and stem color regulation.

Integrated Transcriptomic and Metabolomic Analysis Revealed Regulatory Mechanisms on Flavonoids Biosynthesis in the Skin of Passion Fruit (Passiflora spp.).

Passion fruit is one of the most famous fruit crops in tropical and subtropical regions due to its high edible, medicinal, and ornamental value. Flavonoids, a class of plant secondary metabolites, have important health-related roles. In this study, a total of 151 flavonoid metabolites were identified, of which 25 key metabolites may be the main contributors to the purple phenotype. Using RNA sequencing, 11,180 differentially expressed genes (DEGs) were identified. Among these, 48 flavonoid biosynthesis genes (PAL, 4CL, C4H, CHS, CHI, F3H, DFR, ANS, and UFGT) and 123 transcription factors were identified. Furthermore, 12 distinct modules were identified through weighted gene coexpression network analysis, of which the brown module displays a robust positive correlation with numerous flavonoid metabolites. Overexpression of PeMYB114 significantly promoted flavonoids accumulation in tobacco leaves. Our study provided a key candidate gene for molecular breeding to improve color traits in passion fruit.

Metabolomic and Transcriptomic Analysis Reveals Flavonoid-Mediated Regulation of Seed Antioxidant Properties in Peanut Seed Vigor.

Peanut (Arachis hypogaea L.) is an oilseed crop grown worldwide. Flavonoids have profound benefits for plant growth and development because of their powerful antioxidant properties. Seed vigor is an important indicator of seed quality. However, how flavonoids impact seed vigor formation in large-seed peanuts is still poorly understood. Here, we profiled flavonoids, phytohormones, and transcriptomes of developing seeds of large-seed peanut varieties with low (ZP06) and high (H8107) seed vigor. A total of 165 flavonoids were identified, 51 of which were differentially accumulated in ZP06 and H8107. Lower levels of dihydromyricetin (0.28 times) and hesperetin-7-O-glucoside (0.26 times) were observed in ZP06 seeds than in H8107. All flavonoid biosynthesis structural genes were down-regulated in ZP06. The different hormone levels found in ZP06 and H8107 seeds could be associated with the expression of flavonoid biosynthesis genes via MYB and bHLH transcription factors. Dihydromyricetin could relate to ZP06's poor seed vigor by impacting its seed antioxidant properties. Thus, the presence of flavonoids in large-seed peanuts could contribute to their physiological quality and germination potential through controlling the accumulation of reactive oxygen species to improve seed antioxidant properties.

Tissue-specific transcriptome analyses unveils candidate genes for flavonoid biosynthesis, regulation and transport in the medicinal plant Ilex asprella

It is not clear that the genes involved with flavonoids synthesis, regulation and transport in Ilex asprella. Transcriptome analysis of leaf, stem and root has uncovered 28,478 differentially expressed genes (DEGs) that are involved in various biological processes. Among these, the expression of 31 candidate synthetase genes, 19 transcription factors, and 5 transporters associated with flavonoid biosynthesis varies across tissues, encompassing seven complete biosynthetic pathways (stilbene, aurone, flavone, isoflavone, flavonol, phlobaphene, and anthocyanin) and one partial pathway (proanthocyanidin). Tissue-specific expression patterns suggest that the stilbenes, aurones, flavones and anthocyanin branches are more prominent in roots, as indicated by key genes such as STS(Ilex_044726), CH4ʹGT(Ilex_047989), FNS(Ilex_043640) and UFGT(Ilex_014720). In leaves, the phlobaphenes and flavonols branches are dominant, determined by CHI(Ilex_005941), FNR(Ilex_039777) and FLS(Ilex_046424). The isoflavone pathway appears to be more active in stems due to the presence of IFS(Ilex_029360), mirroring the accumulation of the intermediate metabolite chalcone, which is regulated by CHS(Ilex_047537). The absence of LAR genes implies that gallocatechin, and catechin liked proanthocyanidins cannot be synthesized in I. asprella. Meanwhile, the general phenylpropanoid pathway is more active in roots, stems than in leaves, as evidenced by the expression of PAL(Ilex_042231, Ilex_014816), C4H(Ilex_017598), and 4CL(Ilex_042033). Flavanone, dihydroflavonol and leucoanthocyanidin, key intermediates, accumulate more rapidly in stem, stem and root, respectively, regulated by CHI(Ilex_005941), F3H(Ilex_004635) and DFR(Ilex_004771). Correlation and network analyses reveal that candidate regulators and transporters are closely associated with the synthesis genes. The study provides profound snoop into flavonoids metabolism in I. asprella and offers valuable refer for medicinal plant.

Manipulating flavonoid biosynthesis in Trigonella persica through controlled spectral lighting

Trigonella persica Boiss., is renowned for its rich phytochemical profile, particularly the presence of the flavonol quercetin. This study explored the effects of various light treatments, including blue, red, blue-red (1:1) radiation (BRR), and pink fluorescent light (PFL), on the biochemical and molecular mechanisms governing quercetin and flavonoid biosynthesis in T. persica at different growth stages. Our results showed that light treatments significantly influenced the activity of key enzymes phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) during germination and vegetative growth, with blue light inducing higher PAL and TAL activities compared to control conditions. High-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses revealed that 48-h-old sprouts grown under red light exhibited the highest levels of flavonoid components and phenolic acids, with catechin as the predominant flavonoid. Notably, BRR treatment led to elevated concentrations of the bioavailable quercetin-3-rhamnoside in 48-h-old sprouts, while 15-day-old plants grown under PFL conditions showed a significant accumulation of the sulfated quercetin-3-sulfate. Real-time PCR analysis demonstrated that BRR upregulated the expression of flavonoid biosynthesis genes PAL, chalcone synthase (CHS), and chalcone isomerase (CHI) in sprouts, whereas PFL treatment induced higher expression of these genes, as well as cinnamate 4-hydroxylase (C4H), in aerial parts. These findings suggest that targeted light treatments, particularly blue and red LED light, can enhance the accumulation of bioavailable quercetin-3-rhamnoside during T. persica germination and sprouts exhibit higher levels of flavonoids and phenolic acids than aerial parts during different vegetative growth stages.

Strigolactones Regulate Secondary Metabolism and Nitrogen/Phosphate Signaling in Tea Plants via Transcriptional Reprogramming and Hormonal Interactions.

Strigolactones (SLs) are known to regulate plant architecture formation, nitrogen (N) and phosphorus (P) responses, and secondary metabolism, but their effects in tea plants remain unclear. We demonstrated that the application of a bioactive SL analogue GR24 either to tea roots or leaves initially stimulated but later inhibited catechins, theanine, and caffeine biosynthesis. GR24 treatment also promoted the accumulation of flavonols and insoluble proanthocyanidins in a time- and dose-dependent manner. GR24 influenced flavonoid and theanine biosynthesis genes, such as up-regulating CsTT2c, CsMYB12, and CsbZIP1, modulating N-responsive and assimilation genes (CsNRT1,1, CsGSI/TS1, CsHRS1, CsPHR1, CsNLA1, and CsLBD37/38/39), and repressing N/P transport and signaling genes (CsPHO2, CsPHT1s, CsNRT2,2, CsHHO1, and CsWRKY38). GR24-induced changes in secondary metabolites were also observed in the leaves of tea plants. GR24-regulated CsLBD37a interacted with CsTT8a and CsTT2c, repressing catechins biosynthesis by interrupting MBW complex formation. GR24 regulated caffeine biosynthesis and regulator genes CsS40 and CsNAC7 and may thereby suppress caffeine production. GR24 altered the transcriptomic profiles of multiple hormone biosynthesis and signaling genes that potentially regulate tea characteristic metabolism and N/P signaling. This study provides new insights into SL-induced transcriptional reprogramming that leads to changes in N/P nutrition, secondary metabolism, and hormone signaling in tea plants.

Metabolome and transcriptome reveal the biosynthesis of flavonoids and amino acids in Isatis indigotica fruit during development.

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.

Identification and analysis of CmOMT gene family in Chrysanthemum morifolium

Chrysanthemum morifolium is rich in hydroxyflavonoids and methoxyflavonoids(OMFs), and dissecting the biosynthetic pathway of OMFs in Ch. morifolium is of great theoretical and economic value because of the diverse physiological activities and pharmacological effects of OMFs. To investigate the biosynthetic pathway of OMFs in Ch. morifolium, this study systematically analyzed the CmOMT gene family based on the whole genome data of Ch. morifolium by using bioinformatics approaches. The results showed that 123 CmOMT genes were identified in the genome of Ch. morifolium, encoding 112-541 amino acid residues. The putative proteins had the isoelectric points ranging from 4.92 to 7.59, relative molecular mass ranging from 12 651.69 to 61 300.23. Except CmOMT50, the rest 122 proteins were acidic and 79 of them were hydrophilic. CmOMTs predominantly existed in the cytoplasm with uneven distribution. Seven pathways were enriched by the Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis, including metabolism, phenylpropanoid biosynthesis, and isoflavonoid biosynthesis. The Gene Ontology(GO) enrichment results showed that the 123 CmOMT genes were annotated to 42 GO terms of 12 molecular functions(such as O-methyltransferase activity and S-adenosylme-thanethionine-dependent methyltransferase activity) and 27 biological processes(such as methylation and flavonoid metabolism processes). The prediction results of the cis-acting elements in the promoter region suggested that the CmOMT gene family was involved in the responses to multiple biotic and abiotic stress conditions including light, drought, low temperature, methyl jasmonate, and abscisic acid. In addition, the promoters of CmOMT genes had MYB binding sites involved in the regulation of genes for flavonoid biosynthesis. The analysis of the CmOMT gene family provides a reference for deciphering the biosynthetic pathway of OMFs in Ch. morifolium and lays a foundation of the subsequent mining and functional validation of candidate OMT genes in C. morifolium.

Metabolite concentrations and the expression profiles of the corresponding metabolic pathway genes in eggplant (Solanum melongena L.) fruits of contrasting colors.

Eggplant (Solanum melongena L.) ranks fifth in importance among vegetable crops of the Solanaceae family, in part due to the high antioxidant properties and polyphenol content of the fruit. Along with the popular purple-fruited varieties of S. melongena, there are cultivars, the fruits of which are rich in phenolic compounds, but are white-colored due to the lack of anthocyanin biosynthesis. Determination of the amount of anthocyanins and other phenolic compounds, as well as carotenoids and sugars, is included in the assessment of the quality of eggplant fruits of commercial (technical) ripeness. In addition to antioxidant and taste properties, these metabolites are associated with fruit resistance to various stress factors. In this study, a comparative analysis of the content of anthocyanins, carotenoids and soluble sugars (sucrose, glucose, fructose) in the peel and pulp of the fruit of both technical and biological ripeness was carried out in purple-fruited (cv. Vlas) and white-fruited (cv. Snezhny) eggplant accessions of domestic selection. The peel and pulp of biologically ripe fruits of the cvs Vlas and Snezhny were used for comparative transcriptomic analysis. The key genes of the flavonoid and carotenoid metabolism, sucrose hydrolysis, and soluble sugar transport were shown to be differentially expressed between fruit tissues, both within each cultivar and between them. It has been confirmed that the purple color of the peel of the cv. Vlas fruit is due to substantial amounts of anthocyanins. Flavonoid biosynthesis genes showed a significantly lower expression level in the ripe fruit of the cv. Vlas in comparison with the cv. Snezhny. However, in both cultivars, transcripts of anthocyanin biosynthesis genes (DFR, ANS, UFGT) were not detected. Additionally, the purple fruit of the cv. Vlas accumulated more carotenoids and sucrose and less glucose and fructose than the white fruit of the cv. Snezhny. Biochemical data corresponded to the differential expression pattern of the key genes encoding the structural proteins of metabolism and transport of the compounds analyzed.

Duplication and sub-functionalization of flavonoid biosynthesis genes plays important role in Leguminosae root nodule symbiosis evolution.

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.

Functional impacts of PtrMYB203 on phenylpropanoid pathway regulation and wood properties in hybrid poplar

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.

Integrative Transcriptomic and Metabolic Analyses Reveal That Flavonoid Biosynthesis Is the Key Pathway Regulating Pigment Deposition in Naturally Brown Cotton Fibers.

Brown cotton is a major cultivar of naturally colored cotton, and brown cotton fibers (BCFs) are widely utilized as raw materials for textile industry production due to their advantages of being green and dyeing-pollution-free. However, the mechanisms underlying the pigmentation in fibers are still poorly understood, which significantly limits their extensive applications in related fields. In this study, we conducted a multidimensional comparative analysis of the transcriptomes and metabolomes between brown and white fibers at different developmental periods to identify the key genes and pathways regulating the pigment deposition. The transcriptomic results indicated that the pathways of flavonoid biosynthesis and phenylpropanoid biosynthesis were significantly enriched regulatory pathways, especially in the late development periods of fiber pigmentation; furthermore, the genes distributed in the pathways of PAL, CHS, F3H, DFR, ANR, and UFGT were identified as significantly up-regulated genes. The metabolic results showed that six metabolites, namely (-)-Epigallocatechin, Apiin, Cyanidin-3-O-glucoside, Gallocatechin, Myricetin, and Poncirin, were significantly accumulated in brown fibers but not in white fibers. Integrative analysis of the transcriptomic and metabolomic data demonstrated a possible regulatory network potentially regulating the pigment deposition, in which three MYB transcription factors promote the expression levels of flavonoid biosynthesis genes, thereby inducing the content increase in (-)-Epigallocatechin, Cyanidin-3-O-glucoside, Gallocatechin, and Myricetin in BCFs. Our findings provide new insights into the pigment deposition mechanism in BCFs and offer references for genetic engineering and breeding of colored cotton materials.

Physiological and metabolomic analyses reveal the resistance response mechanism to tea aphid infestation in new shoots of tea plants (Camellia sinensis)

The tea plant (Camellia sinensis (L.) O. Kuntze) is an important woody economic crop and often attacked by various insect pests such as tea aphids (Toxoptera aurantii Boyer de Fonscolombe). To reveal tea plant responses to tea aphid infestation, tea aphid-resistant cultivar ‘Taicha13’ and -susceptible one ‘Jingfeng’ were first screened, and then widely targeted metabolomics profiling, biochemical parameter and RT-qPCR analyses were performed in three samples, TCN (tea aphid non-infested ‘Taicha13’), JFN (tea aphid non-infested ‘Jingfeng’) and JFA (tea aphid-infested ‘Jingfeng’). The results showed amnio acid (plus proline) and protein content, and amnio acid: sucrose ratio were significantly less abundant in TCN relative to JFN, suggesting TCN is low nutritional quality for tea aphids. Metabolite profiling revealed that flavonoids and JA-isoleucine (JA-Ile) were more enriched in TCN and JFA compared to JFN, proposing effective roles of flavonoids and JA-Ile in tea plant defense against tea aphid infestation. RT-qPCR results demonstrated that five JA biosynthesis genes (CsLOX2, CsLOX3, CsOPR, CsAOC and CsJAR) in exception of CsAOS, and six flavonoid biosynthesis genes (CsPAL, CsC4H, CsCHS1, CsCHS2, CsFL and CsDFR) shared similarly higher abundance in TCN than in JFN, and exhibited elevated expression levels in JFA relative to JFN, further suggesting flavonoids and JA confer tea plant resistance against tea aphid feeding. Overall, our findings uncovered that tea aphid-infestation activated defensive responses and highlighted involvement of nutritional quality, flavonoids and JA-Ile in tea plant resistance against tea aphid infestation.

Flavonoid Biosynthesis in Scutellaria baicalensis Georgi: Metabolomics and Transcriptomics Analysis

Scutellaria baicalensis Georgi (SB), a plant of the Lamiaceae family, contains flavonoids with potent human health benefits. The full mechanistic details and regulatory networks related to the biosynthesis of these compounds in SB have been the focus of recent research but are still fragmented. Similarly, a complete account of the metabolites produced, specifically flavonoids, and their distribution in different parts of the plant is incomplete. To provide a more complete picture, herein we have explored the SB metabolites and differentially expressed genes in underground and aerial tissues. Of the 947 metabolites identified, 373 were differentially accumulated flavonoids (DAFs), and 147 of these were differentially accumulated in roots relative to other tissues. Interestingly, roots accumulated more baicalin and baicalein than aboveground tissues, but they were low in scutellarein and wogonoside, in contrast to previous reports. These differences may be attributed to either plant variety, age of the plants, or the extraction protocol. Transcriptomics analysis identified 56 key genes from the flavonoid synthesis pathway in all six SB plant tissues. A weighted gene correlation network analysis conducted using four DAFs (baicalin, baicalein, scutellarein and wogonoside) produced 13 modules. Baicalin and baicalein were positively correlated with one of these modules, whereas wogonoside and scutellarein were correlated with three other modules. Gene expression in these modules was consistent with the observed accumulation of these compounds in plant tissues. Fourteen structural genes were highly correlated with baicalin, baicalein and scutellarein, and 241 transcription factors (TFs) associated to these four compounds. The 13 highly correlated structural genes and 21 highly correlated TFs were used to construct correlation networks, where genes were identified to be highly correlated with flavonoid biosynthesis genes. Overexpression of some of these genes, namely, SbMYB8 (Sb02g25620), SbMYB14 (Sb09g00160) and SbbHLH94 (Sb07g11990), in SB callus increased flavonoid content and regulated the expression of genes involved in the flavonoid biosynthesis pathway, confirming their association to flavonoid production. Overall, the present work contributes to delineating the differences in flavonoid biosynthesis among different SB tissues.

Adaptive significance and origin of flavonoid biosynthesis genes in the grain of cultivated cereals

The majority of cultivated cereals including maize, rice, wheat, barley, oat and rye are consisted of numerous varieties lacking anthocyanin pigmentation or having weak coloration of vegetative organs and/or caryopses. Only rare local races and wild related species have intense coloration of plants and/or grains. The coloration of caryopses is associated with the biosynthesis of colored flavonoids in maternal (pericarp and testa) and hybrid (aleuron) caryopsis tissues. The trait is controlled by dominant alleles of regulatory genes encoding conserved transcription factors of the MYB, bHLH-MYC, and WD40 families forming the MBW protein complex. Recent studies have proven the participation of uncolored and colored flavonoids in the response of plants to biotic and abiotic stresses, and significance of their presence in the whole grain foods has been determined. However, many questions about the adaptive effects and health benefits of anthocyanins remain unanswered. In particular, the reasons why the dominant alleles of regulatory genes controlling pericarp coloration did not become widespread in the course of domestication and breeding of cereals are not clear, although these genes receive special attention in association with health-improving effects of grain nutrition. This article discusses the similarity and specificity of the genetic control of the biosynthesis of flavonoids in the caryopsis in three related cultivated cereals – wheat, barley and rye, and their biological role in the development of the caryopsis and seed germination.

Metabolic and Transcriptomic Profile Revealing the Differential Accumulating Mechanism in Different Parts of Dendrobium nobile.

Dendrobium nobile is an important orchid plant that has been used as a traditional herb for many years. For the further pharmaceutical development of this resource, a combined transcriptome and metabolome analysis was performed in different parts of D. nobile. First, saccharides, organic acids, amino acids and their derivatives, and alkaloids were the main substances identified in D. nobile. Amino acids and their derivatives and flavonoids accumulated strongly in flowers; saccharides and phenols accumulated strongly in flowers and fruits; alkaloids accumulated strongly in leaves and flowers; and a nucleotide and its derivatives and organic acids accumulated strongly in leaves, flowers, and fruits. Simultaneously, genes for lipid metabolism, terpenoid biosynthesis, and alkaloid biosynthesis were highly expressed in the flowers; genes for phenylpropanoids biosynthesis and flavonoid biosynthesis were highly expressed in the roots; and genes for other metabolisms were highly expressed in the leaves. Furthermore, different members of metabolic enzyme families like cytochrome P450 and 4-coumarate-coA ligase showed differential effects on tissue-specific metabolic accumulation. Members of transcription factor families like AP2-EREBP, bHLH, NAC, MADS, and MYB participated widely in differential accumulation. ATP-binding cassette transporters and some other transporters also showed positive effects on tissue-specific metabolic accumulation. These results systematically elucidated the molecular mechanism of differential accumulation in different parts of D. nobile and enriched the library of specialized metabolic products and promising candidate genes.

MYB4, a member of R2R3-subfamily of MYB transcription factor functions as a repressor of key genes involved in flavonoid biosynthesis and repair of UV-B induced DNA double strand breaks in Arabidopsis

Plants accumulate flavonoids as part of UV-B acclimation, while a high level of UV-B irradiation induces DNA damage and leads to genome instability. Here, we show that MYB4, a member of the R2R3-subfamily of MYB transcription factor plays important role in regulating plant response to UV-B exposure through the direct repression of the key genes involved in flavonoids biosynthesis and repair of DNA double-strand breaks (DSBs). Our results demonstrate that MYB4 inhibits seed germination and seedling establishment in Arabidopsis following UV-B exposure. Phenotype analyses of atmyb4-1 single mutant line along with uvr8-6/atmyb4-1, cop1-6/atmyb4-1, and hy5-215/atmyb4-1 double mutants indicate that MYB4 functions downstream of UVR8 mediated signaling pathway and negatively affects UV-B acclimation and cotyledon expansion. Our results indicate that MYB4 acts as transcriptional repressor of two key flavonoid biosynthesis genes, including 4CL and FLS, via directly binding to their promoter, thus reducing flavonoid accumulation. On the other hand, AtMYB4 overexpression leads to higher accumulation level of DSBs along with repressed expression of several key DSB repair genes, including AtATM, AtKU70, AtLIG4, AtXRCC4, AtBRCA1, AtSOG1, AtRAD51, and AtRAD54, respectively. Our results further suggest that MYB4 protein represses the expression of two crucial DSB repair genes, AtKU70 and AtXRCC4 through direct binding with their promoters. Together, our results indicate that MYB4 functions as an important coordinator to regulate plant response to UV-B through transcriptional regulation of key genes involved in flavonoids biosynthesis and repair of UV-B induced DNA damage.

In silico modelling of proteins encoded by important genes of flavonoid biosynthesis pathway in Gymnema sylvestre R. Br.

Understanding the proteins and metabolites produced by plants aids in developing superior planting materials. Gymnema sylvestre, a significant medicinal plant containing beneficial compounds such as oleanane, dammarane, flavonoids and gymnemic acids, exhibiting antioxidant and antidiabetic properties, making it a promising alternative treatment for Type 2 diabetes with minimal side effects. Transcriptomics in G. sylvestre yielded Coding DNA Sequences (CDS) for 12 key genes of flavonoid biosynthesis. Translated protein sequences of those 12 genes underwent characterization using protPARAM and i-TASSER. In silico protein modelling found similarities to validated enzymes, suggesting functional potential of the studied proteins. The study enlightens the first comprehensive insight into key proteins involved in G. sylvestre's flavonoid biosynthesis pathway. GsCHS (G. sylvestre chalcone synthase), a key enzyme in flavonoid biosynthesis, corroborated its phylogenetic proximity to Vincetoxicum mongolicum's CHS. Protein docking analysis delineated the distinctive features of GsCHS from cpCHS1 of Cyclosorus parasiticus, particularly in its substrate binding pockets and interactive sites crucial for synthesizing essential flavonoids.

Integrated transcriptomics and metabolomics revealed the mechanism of catechin biosynthesis in response to lead stress in tung tree (Vernicia fordii)

Lead (Pb) affects gene transcription, metabolite biosynthesis and growth in plants. The tung tree (Vernicia fordii) is highly adaptive to adversity, whereas the mechanisms underlying its response to Pb remain uncertain. In this work, transcriptomic and metabolomic analyses were employed to study tung trees under Pb stress. The results showed that the biomass of tung seedlings decreased with increasing Pb doses, and excessive Pb doses resulted in leaf wilting, root rot, and disruption of Pb homeostasis. Under non-excessive Pb stress, a significant change in the expression patterns of flavonoid biosynthesis genes was observed in the roots of tung seedlings, leading to changes in the accumulation of flavonoids in the roots, especially the upregulation of catechins, which can chelate Pb and reduce its toxicity in plants. In addition, Pb-stressed roots showed a large accumulation of VfWRKY55, VfWRKY75, and VfLRR1 transcripts, which were shown to be involved in the flavonoid biosynthesis pathway by gene module analysis. Overexpression of VfWRKY55, VfWRKY75, and VfLRR1 significantly increased catechin concentrations in tung roots, respectively. These data indicate that Pb stress-induced changes in the expression patterns of those genes regulate the accumulation of catechins. Our findings will help to clarify the molecular mechanism of Pb response in plants.