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Nitrogen deficiency is a key constraint on crop yield. Cassava, the world's sixth-largest food crop and a crucial source of feed and industrial materials, can thrive in marginal soils, yet its yield is still significantly affected by limited nitrogen availability. Investigating cassava's response mechanisms to nitrogen scarcity is therefore essential for advancing molecular breeding and identifying nitrogen-efficient varieties. This research undertook a comprehensive analysis of cassava seedlings' physiological, gene expression, and metabolite responses under low nitrogen stress. Findings revealed that nitrogen deficiency drastically suppressed seedling growth, significantly reduced nitrate and ammonium transport to aerial parts, and led to a marked increase in carbohydrate, reactive oxygen species, and ammonium ion levels in the leaves. Transcriptomic and metabolomic analyses further demonstrated notable alterations in genes and metabolites linked to carbon and nitrogen metabolism, flavonoid biosynthesis, and the purine metabolic pathway. Additionally, several transcription factors associated with cassava flavonoid biosynthesis under nitrogen-deficient conditions were identified. Overall, this study offers fresh insights and valuable genetic resources for unraveling cassava's adaptive mechanisms to nitrogen deprivation. [ABSTRACT FROM AUTHOR]

Safflower (Carthamus tinctorius L.), a versatile medicinal and economic crop, harbors untapped genetic resources essential for stress resilience and metabolic regulation. The TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factors, exclusive to plants, are pivotal in orchestrating growth, development, and stress responses, yet their roles in safflower remain unexplored. Here, we report the comprehensive identification and characterization of 26 safflower TCP genes (CtTCPs), categorized into Class I (PROLIFERATING CELL FACTOR, PCF) and Class II (CINCINNATA and TEOSINTE BRANCHED1/CYCLOIDEA, CIN and CYC/TB1) subfamilies. Comparative phylogenetics, conserved motif, and gene structure analyses revealed a high degree of evolutionary conservation and functional divergence within the gene family. Promoter analyses uncovered light-, hormone-, and stress-responsive cis-elements, underscoring their regulatory potential. Functional insights from qRT-PCR analyses demonstrated dynamic CtTCP expression under abiotic stresses, including abscisic acid (ABA), Methyl Jasmonate (MeJA), Cold, and ultraviolet radiation b (UV-B) treatments. Notably, ABA stress triggered a significant increase in flavonoid accumulation, correlated with the upregulation of key flavonoid biosynthesis genes and select CtTCPs. These findings illuminate the complex regulatory networks underlying safflower's abiotic stress responses and secondary metabolism, offering a molecular framework to enhance crop resilience and metabolic engineering for sustainable agriculture [ABSTRACT FROM AUTHOR]

Introduction: Introduction: Light is not only essential for plant photosynthesis and growth, but also acts as a signal to regulate its secondary metabolism. Despite the influence of light quality on the yield and flavonoid compounds in commercial crops is well-documented, its role in regulating wild understorey species, particularly medicine plants whose flavonoid biosynthesis driven by multiple spectral regions of canopy sunlight, is less understood. Methods: To address it, we conducted a light-quality manipulation experiment on Scutellaria baicalensis Georgi, a widespread understorey medicinal species, with light-emitting diodes (LED). This study included eight treatments: UV-A (UV-A radiation), CK (control group), Green (monochromatic green light), and different combinations of blue and red light (R0B4: monochromatic blue light; R1B3: 25% Red+75% Blue light; R1B1: 50% Red+50% Blue light; R3B1: 75% Red+25% Blue light; R4B0: monochromatic red light). Results: Our results showed that light quality significantly drove morphology, biomass accumulation, and flavonoids biosynthesis in S. baicalensis. R0B4 treatment promoted growth and flavonoids accumulation, including baicalin, and wogonoside concentrations. In contrast, UV-A radiation and green light negatively impacted these parameters compared to CK treatment. Interestingly, plant biomass and flavonoid concentrations were lower in R1B3, R1B1 and R3B1 treatments compared to monochromatic blue or red light. Discussion: Our study found that red light may antagonize blue light-stimulated growth and flavonoids accumulation, indicating a complex crosstalk between photoreceptors. These findings highlight the importance of blue light for optimizing the yield and quality of S. baicalensis in the understorey cultivation. It provides practice suggestion for the efficient management and sustainable cultivation of understorey medicinal plants. [ABSTRACT FROM AUTHOR]

Background: Safflower thrives in dry environments but faces difficulties with flowering in wet and rainy summers. Flavonoids play a role in flower development and can potentially alleviate these challenges. Furthermore, the FLOWERING LOCUS T (FT) family of phosphatidylethanolamine-binding protein (PEBP) genes play a crucial role in the photoperiodic flowering pathway. However, their direct impact on flowering and flavonoid biosynthesis under different light duration is elusive. Results: Utilizing the genome sequencing of Safflower (Jihong NO.1), the current study identifies three specific genes (CtFT1, CtFT2, and CtFT3) that exhibit upregulation in response to long-day conditions. The overexpression of CtFT2, displayed an early, whereas CtFT1 and CtFT3 late flowering phenotype in Arabidopsis thaliana. Interestingly, the transient overexpression of CtFT1 in safflower leaves caused early flowering, while overexpressing CtFT2 and CtFT3 led to late flowering. Additionally, overexpressing CtFT3 in Arabidopsis and CtFT1, CtFT2, and CtFT3 in safflower leaves, significantly increased flavonoid synthesis. Conclusions: These findings showed that overexpression of CtFT genes could affect the flowering time and significantly increase the flavonoid content of safflower. The function of CtFT gene is different in safflower and Arabidopsis. This study provides valuable insights into the role of CtFT genes in flower formation and flavonoid synthesis in safflower, which may help in improving safflower breeding quality and its adaptability to diverse environmental conditions. [ABSTRACT FROM AUTHOR]

Elevated levels of reactive oxygen species (ROS) are caused by ultraviolet B radiation (UV-B) stress. In response, plants strengthen their cell membranes, impeding photosynthesis. Additionally, UV-B stress initiates oxidative stress within the antioxidant defense system and alters secondary metabolism, particularly by increasing the quantity of UV-absorbing compounds such as flavonoids. The v-myb avian myeloblastosis viral oncogene homolog (MYB) transcription factor (TF) may participate in a plant's response to UV-B damage through its regulation of flavonoid biosynthesis. In this study, we discovered that the photosynthetic activity of Rhododendron chrysanthum Pall. (R. chrysanthum) decreased when assessing parameters of chlorophyll (PSII) fluorescence parameters under UV-B stress. Concurrently, antioxidant system enzyme expression increased under UV-B exposure. A multi-omics data analysis revealed that acetylation at the K68 site of the RcTRP5 (telomeric repeat binding protein of Rhododendron chrysanthum Pall.) transcription factor was upregulated. This acetylation modification of RcTRP5 activates the antioxidant enzyme system, leading to elevated expression levels of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT). Upregulation is also observed at the K95 site of the chalcone isomerase (CHI) enzyme and the K178 site of the anthocyanidin synthase (ANS) enzyme. We hypothesize that RcTRP5 influences acetylation modifications of CHI and ANS in flavonoid biosynthesis, thereby indirectly regulating flavonoid production. This study demonstrates that R. chrysanthum can be protected from UV-B stress by accumulating flavonoids. This could serve as a useful strategy for enhancing the plant's flavonoid content and provide a valuable reference for research on the metabolic regulation mechanisms of other secondary substances. [ABSTRACT FROM AUTHOR]

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

Antioxidant activity and flavonoid content are important characteristics of colored rice grains. Previously, we obtained a preliminary understanding of the metabolic markers of antioxidant activity, namely, phlorizin and trilobatin, in different colored rice varieties and purple rice grains at different growth stages, but the mechanisms associated with these markers have not yet been confirmed. In this study, purple rice was selected as the experimental material, and clover extract was applied during the grain-filling stage to explore the impact of clover extract on the total antioxidant capacity and flavonoid biosynthesis in purple rice grains. The results indicated that the total flavonoid content, total phenolic content, oligomeric proanthocyanidin content, and total antioxidant capacity of purple rice grains treated with an exogenous application of trilobatin (T30) were significantly greater than those of the control (CK). The flavonoids in the T30 and CK groups accounted for 29.81% of the total flavonoids. The phenylpropanoid biosynthesis and flavonoid biosynthesis metabolic pathways were constructed on the basis of the differentially abundant metabolites between the T30 and CK groups. Additionally, 1-O-sinapoyl-beta-D-glucose, coniferaldehyde, 6″-acetylapiin, and kaempferol-3-O-rutinoside were determined to be essential metabolites for trilobatin-mediated flavonoid biosynthesis in purple rice. The correlation network diagram between biochemical indexes and metabolites revealed that 1-O-sinapoyl-beta-D-glucose, coniferaldehyde, 6″-acetylapiin, and kaempferol-3-O-rutinoside were important metabolites. This study provides a scientific basis for improving the nutritional quality of rice grains and understanding the flavonoid biosynthesis pathway. [ABSTRACT FROM AUTHOR]

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.

Abstract The golden camellia, a highly valued ornamental plant, belongs to the Chrysantha section, the sole group in the Camellia genus that produces yellow flowers. The cultivation and broader application of this unique species have been significantly hindered by the scarcity of golden camellia and the limited knowledge of the molecular mechanisms governing yellow flower development. To investigate the dynamics of metabolite accumulation and gene expression during the yellow flower formation of C. perpetua, a continuously flowering golden camellia, a comprehensive metabolomic and transcriptomic analysis was performed across five distinct developmental stages. This analysis identified 1,160 metabolites, with flavonoids constituting the largest proportion at 21.6%. Comparative transcriptomic profiling indicated that differentially expressed genes (DEGs) associated with photosynthesis were predominantly active during the early stages of flower development, while DEGs involved in the flavonoid biosynthesis pathway showed a marked increase as the flowers matured. The integrated metabolomic and transcriptomic data highlighted the pivotal regulatory role of the flavonoid biosynthesis pathway in the formation of yellow flowers in C. perpetua, identifying 50 key genes and 17 crucial metabolites and mapping their interaction networks. Furthermore, weighted gene co-expression network analysis pinpointed several hub genes, such as flavonol synthase, which are likely critical to flavonoid production due to their elevated expression levels, particularly in the early bud and yellowing phases. This study provides valuable insights into the molecular pathways that underlie yellow flower development in golden camellia species.

Anthocyanins, flavonoid pigments, are responsible for the purple and red hues in potato tubers. This study analyzed tubers from four potato cultivars—red RR, purple HJG, yellow QS9, and white JZS8—to elucidate the genetic mechanisms underlying tuber pigmentation. Our transcriptomic analysis identified over 2400 differentially expressed genes between these varieties. Notably, genes within the flavonoid biosynthesis pathway were enriched in HJG and RR compared to the non-pigmented JZS8, correlating with their higher levels of anthocyanin precursors and related substances. Hierarchical clustering revealed inverse expression patterns for the key genes involved in anthocyanin metabolism between pigmented and non-pigmented varieties. Among these, several MYB transcription factors displayed strong co-expression with anthocyanin biosynthetic genes, suggesting a regulatory role. Specifically, the expression of 16 MYB genes was validated using qRT-PCR to be markedly higher in pigmented HJG and RR versus JZS8, suggesting that these MYB genes might be involved in tuber pigmentation. This study comprehensively analyzed the transcriptome of diverse potato cultivars, highlighting specific genes and metabolic pathways involved in tuber pigmentation. These findings provide potential molecular targets for breeding programs focused on enhancing tuber color.

Hyperaccumulators within the Noccaea genus possess many promising genetic and metabolic adaptations that could be potentially exploited to support phytoremediation efforts and/or crop improvement and biofortification. Although hyperaccumulation is very common in this genus, individual species display specific traits as they can accumulate different elements (e.g., zinc, cadmium, and/or nickel). Moreover, there appears to be some populational variability with natural selection increasing the metal tolerance in metallicolous populations. Therefore, employing robust methods, such as integrated analysis of the transcriptome and metabolome, is crucial for uncovering pivotal candidate genes and pathways orchestrating the response to metal stress in Noccaea hyperaccumulators. Our study highlights several species-specific traits linked to the detoxification of metals and metal-induced oxidative stress in hyperaccumulating N. praecox when compared to a closely related model species, N. caerulescens, when grown in the field. Transcriptome analysis revealed distinct differences between the three studied natural Noccaea populations. Notably, we observed several pathways frequently connected to metal stress, i.e., glutathione metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis, which were enriched. These differences were observed despite the relative evolutionary closeness of studied species, which emphasizes the importance of further expanding our knowledge on hyperaccumulators if we want to exploit their mechanisms for phytoremediation efforts or food quality improvements. [ABSTRACT FROM AUTHOR]

The basic helix-loop-helix (bHLH) transcription factors play crucial roles in various processes, such as plant development, secondary metabolism, and response to biotic/abiotic stresses. Sophora flavescens is a widely used traditional herbal medicine in clinical practice, known for its abundant flavonoids as the main active compounds. However, there has been no comprehensive analysis of S. flavescens bHLH (SfbHLH) gene family reported currently. In this study, we identified 167 SfbHLH genes and classified them into 23 subfamilies based on comparative genomics and phylogenetic analysis. Furthermore, widespread duplications significantly contributed to the expansion of SfbHLH family. Notably, SfbHLH042 was found to occupy a central position in the bHLH protein-protein interaction network. Transcriptome analysis of four tissues (leaf, stem, root and flower) revealed that most SfbHLH genes exhibited high expression levels exclusively in specific tissues of S. flavescens. The integrated analysis of transcriptomics and metabolomics during pod development stages revealed that SfbHLH042 may play a central role in connecting SfbHLH genes, flavonoids, and key enzymes involved in the biosynthesis pathway. Moreover, we also checked the expression of 8 SfbHLH genes using RT-qPCR analysis to realize the expression profiles of these genes among various tissues at different cultivated periods and root development. Our study would aid to understand the phylogeny and expression profile of SfbHLH family genes, and provide a promising candidate gene, SfbHLH042 , for regulating the biosynthesis of flavonoids in S. flavescens. [ABSTRACT FROM AUTHOR]

Seed vigor is an important trait closely related to improved seed quality and long-term germplasm conservation, and it gradually decreases during storage, which has become a major concern for agriculture. However, the underlying regulatory mechanisms of seed vigor loss in terms of genes remain largely unknown in quinoa. Here, two cultivars of quinoa seeds with different storage performance, Longli No.4 (L4) and Longli No.1 (L1), were subjected to transcriptome sequencing to decipher the pathways and genes possibly related to vigor loss under artificial aging. Multispectral imaging features and germination phenotypes showed significantly less seed vigor loss in L1 than in L4, indicating L1 seeds having stronger aging resistance and storability. Totally, 272 and 75 differentially expressed genes (DEGs) were, respectively, identified in L4 and L1 during aging. Transcriptomic analysis further revealed the differences in metabolic pathways, especially, flavonoid biosynthesis, TCA cycle, and terpenoid backbone biosynthesis were significantly enriched in L4 seeds, while carbon metabolism in L1 seeds, which involved key genes such as CHS , CHI , AACT , ENO1 , IDH , NADP-ME , and HAO2L. It indicated that the adverse effects on flavonoids and terpenoids induced by aging might be the significant reasons for more vigor loss in storage sensitive seeds, whereas storage tolerant seeds had a stronger ability to maintain carbon metabolism and energy supply. These findings elucidated the underlying molecular mechanism of seed vigor loss in quinoa, which also provided novel insights into improving seed vigor through modern molecular breeding strategies. [ABSTRACT FROM AUTHOR]

The basic helix-loop-helix (bHLH) transcription factors play crucial roles in various processes, such as plant development, secondary metabolism, and response to biotic/abiotic stresses. Sophora flavescens is a widely used traditional herbal medicine in clinical practice, known for its abundant flavonoids as the main active compounds. However, there has been no comprehensive analysis of S. flavescens bHLH (SfbHLH) gene family reported currently. In this study, we identified 167 SfbHLH genes and classified them into 23 subfamilies based on comparative genomics and phylogenetic analysis. Furthermore, widespread duplications significantly contributed to the expansion of SfbHLH family. Notably, SfbHLH042 was found to occupy a central position in the bHLH protein-protein interaction network. Transcriptome analysis of four tissues (leaf, stem, root and flower) revealed that most SfbHLH genes exhibited high expression levels exclusively in specific tissues of S. flavescens. The integrated analysis of transcriptomics and metabolomics during pod development stages revealed that SfbHLH042 may play a central role in connecting SfbHLH genes, flavonoids, and key enzymes involved in the biosynthesis pathway. Moreover, we also checked the expression of 8 SfbHLH genes using RT-qPCR analysis to realize the expression profiles of these genes among various tissues at different cultivated periods and root development. Our study would aid to understand the phylogeny and expression profile of SfbHLH family genes, and provide a promising candidate gene, SfbHLH042, for regulating the biosynthesis of flavonoids in S. flavescens. [ABSTRACT FROM AUTHOR]

Seed vigor is an important trait closely related to improved seed quality and long-term germplasm conservation, and it gradually decreases during storage, which has become a major concern for agriculture. However, the underlying regulatory mechanisms of seed vigor loss in terms of genes remain largely unknown in quinoa. Here, two cultivars of quinoa seeds with different storage performance, Longli No.4 (L4) and Longli No.1 (L1), were subjected to transcriptome sequencing to decipher the pathways and genes possibly related to vigor loss under artificial aging. Multispectral imaging features and germination phenotypes showed significantly less seed vigor loss in L1 than in L4, indicating L1 seeds having stronger aging resistance and storability. Totally, 272 and 75 differentially expressed genes (DEGs) were, respectively, identified in L4 and L1 during aging. Transcriptomic analysis further revealed the differences in metabolic pathways, especially, flavonoid biosynthesis, TCA cycle, and terpenoid backbone biosynthesis were significantly enriched in L4 seeds, while carbon metabolism in L1 seeds, which involved key genes such as CHS, CHI, AACT, ENO1, IDH, NADP-ME, and HAO2L. It indicated that the adverse effects on flavonoids and terpenoids induced by aging might be the significant reasons for more vigor loss in storage sensitive seeds, whereas storage tolerant seeds had a stronger ability to maintain carbon metabolism and energy supply. These findings elucidated the underlying molecular mechanism of seed vigor loss in quinoa, which also provided novel insights into improving seed vigor through modern molecular breeding strategies. [ABSTRACT FROM AUTHOR]

Flavonoids are secondary metabolites present in plant organs and tissues. These natural metabolites are the most prevalent and display a wide range of beneficial physiological effects, making them usually intriguing in several scientific fields. Due to their safety for use and protective attributes, including antioxidant, anti‐inflammatory, anticancer, and antimicrobial functions, flavonoids are broadly utilized in foods, pharmaceuticals, and nutraceuticals. However, conventional methods for producing flavonoids, such as plant extraction and chemical synthesis, entailed dangerous substances, and laborious procedures, with low product yield. Recent studies have documented the ability of microorganisms, such as fungi and bacteria, to synthesize adequate amounts of flavonoids. Bacterial biosynthesis of flavonoids from plant biomass is a viable and environmentally friendly technique for producing flavonoids on a larger scale and has recently received much attention. Still, only a few bacteria species, particularly Escherichia coli, have been extensively studied. The most recent developments in bacterial biosynthesis of flavonoids are reviewed and discussed in this article, including their various applications as natural food biocontrol agents. In addition, the challenges currently faced in bacterial flavonoid biosynthesis and possible solutions, including the application of modern biotechnology approaches for developing bacterial strains that could successfully produce flavonoids on an industrial scale, were elucidated. [ABSTRACT FROM AUTHOR]

Tartary buckwheat (Fagopyrum tataricum Gaertn.) is a coarse grain crop rich in flavonoids that are beneficial to human health because they function as anti-inflammatories and provide protection against cardiovascular disease and diabetes. Flavonoid biosynthesis is a complex process, and relatively little is known about the regulatory pathways involved in Tartary buckwheat. Here, we cloned and characterized the FtMYB163 gene from Tartary buckwheat, which encodes a member of the R2R3-MYB transcription factor family. Amino acid sequence and phylogenetic analysis indicate that FtMYB163 is a member of subgroup 7 (SG7) and closely related to FeMYBF1, which regulates flavonol synthesis in common buckwheat (F. esculentum). We demonstrated that FtMYB163 localizes to the nucleus and has transcriptional activity. Expression levels of FtMYB163 in the roots, stems, leaves, flowers, and seeds of F. tataricum were positively correlated with the total flavonoid contents of these tissues. Overexpression of FtMYB163 in transgenic Arabidopsis enhanced the expression of several genes involved in early flavonoid biosynthesis (AtCHS, AtCHI, AtF3H, and AtFLS) and significantly increased the accumulation of several flavonoids, including naringenin chalcone, naringenin-7-O-glucoside, eriodictyol, and eight flavonol compounds. Our findings demonstrate that FtMYB163 positively regulates flavonol biosynthesis by changing the expression of several key genes in flavonoid biosynthetic pathways. [ABSTRACT FROM AUTHOR]

Main conclusions: The albino phenotype of Agave angustifolia Haw. accumulates higher levels of phenylalanine and phenylpropanoids, while the green phenotype has a greater concentration of phenolic compounds. The metabolic consequences of chlorophyll deficiency in plants continue to be a captivating field of research, especially in relation to production of metabolic compounds. This study conducts a thorough analysis of the metabolome in green (G), variegated (V), and albino (A) phenotypes of Agave angustifolia Haw. Specifically, it examines the differences in the accumulation of compounds related to the phenylpropanoid and flavonoid biosynthesis pathways. Methanol extracts of leaf and meristem tissues from the three phenotypes grown in vitro were analyzed using liquid chromatography coupled with quadrupole time-of-flight high-resolution mass spectrometry (UPLC-MS-QTOF) for untargeted metabolomics and triple quadrupole (QqQ) mass spectrometry for targeted metabolomic analyses. By employing these methods, we discovered notable differences in the levels of important metabolites such as L-phenylalanine, 4-hydroxyphenylpyruvic acid, and various flavonoids among the different phenotypes. The results of our study indicate that the A phenotype shows a significant increase in the levels of phenylalanine and phenylpropanoids in both leaf and meristem tissues. This is in contrast to a decrease in flavonoids, suggesting a metabolic reprogramming to compensate for the lack of chlorophyll. Significantly, compounds such as kaempferol-3-O-glucoside and rutin exhibited significant quantitative reduction in the A leaves, suggesting a subtle modification in the production of flavonols and potentially a changed mechanism for antioxidant protection. This study emphasizes the complex metabolic changes in A. angustifolia´s chlorophyll-deficient phenotypes, providing insight into the complex interplay between primary and secondary metabolism in response to chlorophyll deficiency. Our research not only enhances the comprehension of plant metabolism in albino phenotypes but also opens new avenues for exploring the biochemical and genetic basis of such adaptations, with potential biotechnological applications of these distinct plant variants. [ABSTRACT FROM AUTHOR]

WRKY transcription factor (TF) plays a crucial role in plant abiotic stress response, but it is rarely reported in Michelia crassipes. Our studies have found that the transcription factor McWRKY43, a member of the IIc subgroup, is strongly upregulated under cold stress. In this study, we cloned the full length of McWRKY43 to further investigate the function of McWRKY43 in resistance to cold stress and its possible regulatory pathways in M. crassipes. Under cold stress, the seed-germination rate of transgenic tobacco was significantly higher than that of the wild type, and the flavonoid content, antioxidant enzyme activities, and proline content of transgenic tobacco seedlings were significantly increased, which promoted the expression of flavonoid pathway structural genes. In addition, the transient transformation of McWRKY43 in the M. crassipes leaves also found the accumulation of flavonoid content and the transcription level of flavonoid structural genes, especially McLDOX, were significantly increased under cold stress. Yeast one-hybrid (Y1H) assay showed that McWRKY43 could bind to McLDOX promoter, and the transcription expression of McLDOX was promoted by McWRKY43 during cold stress treatment. Overall, our results indicated that McWRKY43 is involved in flavonoid biosynthetic pathway to regulate cold stress tolerance of M. crassipes, providing a basis for molecular mechanism of stress resistance in Michelia. [ABSTRACT FROM AUTHOR]

The Pyrus spp. (pears) are crucial for the fruit industry; however, low spring temperatures can cause frost damage to their reproductive organs, which poses challenges to the final yields. In this study, we evaluated the response of the flowers and young fruits of the ‘Jinguang’ pear cultivar to low temperatures from integrated phenotypic, physiological, and molecular approaches. We found that the flowers were less sensitive to low temperatures than the young fruits, of which their over-cooling points were −5.6°C and −5.0°C, respectively. Transcriptomic data showed that the differentially expressed genes from flowers and young fruits compared to the control conditions were primarily involved in the biosynthesis of flavonoids, phenylalanine, and tyrosine. Further weighted gene co-expression network analysis uncovered the core transcription factors that may be potentially involved in the pear cold resistance, including MYB20, WRKY53, and WRKY30. Our findings provide valuable insights and candidate gene resources for further exploration of the molecular mechanisms underlying cold resistance in pear trees.

Pepper mild mottle virus (PMMoV), a member of the genus Tobamovirus, causes severe damage on pepper worldwide. Despite its impact, the pathogenicity mechanisms of PMMoV and the pepper plant’s response to infection remain poorly understood. Here, we compared the transcriptomic changes in a susceptible pepper inbred line 21C241 with a resistant inbred line 21C385 seedlings, following systemic PMMoV infection using RNA sequencing. Our results revealed that PMMoV induced more pronounced mosaic symptoms and higher viral accumulation levels in the susceptible line 21C241 compared to the resistant line 21C385. We identified 462 and 401 differentially expressed genes (DEGs) in the systemically-infected leaves of the susceptible and resistant lines, respectively, when compared to their healthy counterparts. The majority of these DEGs were involved in photosynthesis and the biosynthesis of secondary metabolites, with 28 DEGs exhibiting distinct expression patterns between the two lines. Notably, the expression level of the chlorophyll a-b binding protein 13 (CAB13) was significantly up-regulated in resistant line 21C385 following PMMoV infection. Functional analysis through silencing of CAB13 in pepper and Nicotiana benthamiana demonstrated a reduction in PMMoV accumulation, suggesting that CAB13 plays a positive role in facilitating PMMoV infection in pepper plants. Taken together, our findings highlight the distinct gene expression profiles between susceptible and resistant pepper lines in response to PMMoV infection and confirm the proviral role of CAB13. This study provides valuable insights into the molecular mechanisms underlying resistance and susceptibility in pepper plants and may inform future strategies for disease management.

Nitrogen deficiency is a key constraint on crop yield. Cassava, the world’s sixth-largest food crop and a crucial source of feed and industrial materials, can thrive in marginal soils, yet its yield is still significantly affected by limited nitrogen availability. Investigating cassava’s response mechanisms to nitrogen scarcity is therefore essential for advancing molecular breeding and identifying nitrogen-efficient varieties. This research undertook a comprehensive analysis of cassava seedlings’ physiological, gene expression, and metabolite responses under low nitrogen stress. Findings revealed that nitrogen deficiency drastically suppressed seedling growth, significantly reduced nitrate and ammonium transport to aerial parts, and led to a marked increase in carbohydrate, reactive oxygen species, and ammonium ion levels in the leaves. Transcriptomic and metabolomic analyses further demonstrated notable alterations in genes and metabolites linked to carbon and nitrogen metabolism, flavonoid biosynthesis, and the purine metabolic pathway. Additionally, several transcription factors associated with cassava flavonoid biosynthesis under nitrogen-deficient conditions were identified. Overall, this study offers fresh insights and valuable genetic resources for unraveling cassava’s adaptive mechanisms to nitrogen deprivation.

Drought is one of the most detrimental stresses that severely constrains plant growth and productivity. Although Chinese chive (Allium tuberosum Rottler) is a vegetable species that is cultivated and consumed worldwide, few studies have investigated how this species responds to drought. In this study, we conducted transcriptomics, metabolomics, and proteomics analyses on chive seedlings exposed to different water availability conditions (mild drought, moderate drought, severe drought, and re-watering) and found that the accumulation of flavonoids in chive leaves was substantially altered under drought stress. Gene co-expression regulatory network analysis, conducted by integrating transcriptome and metabolome data, revealed a chive R2R3-MYB transcription factor, AtuMYB306, as a central regulator of flavonoid synthesis. Overexpression of AtuMYB306 significantly improved osmotic stress tolerance and enhanced flavonoid content in Arabidopsis. We further demonstrated that AtuMYB306 directly binds to the promoters of three flavonoid biosynthetic genes (Atu4CL, AtuF3H, and AtuF3’H) and activates their expression. These results suggest that AtuMYB306 improves drought tolerance in Chinese chive by enhancing flavonoid biosynthesis to scavenge reactive oxygen species (ROS) generated under water-deficit conditions. Thus, our findings provide evidence that AtuMYB306 playing a pivotal role in improving drought resistance in Chinese chive.

IntroductionIntroduction: Light is not only essential for plant photosynthesis and growth, but also acts as a signal to regulate its secondary metabolism. Despite the influence of light quality on the yield and flavonoid compounds in commercial crops is well-documented, its role in regulating wild understorey species, particularly medicine plants whose flavonoid biosynthesis driven by multiple spectral regions of canopy sunlight, is less understood.MethodsTo address it, we conducted a light-quality manipulation experiment on Scutellaria baicalensis Georgi, a widespread understorey medicinal species, with light-emitting diodes (LED). This study included eight treatments: UV-A (UV-A radiation), CK (control group), Green (monochromatic green light), and different combinations of blue and red light (R0B4: monochromatic blue light; R1B3: 25% Red+75% Blue light; R1B1: 50% Red+50% Blue light; R3B1: 75% Red+25% Blue light; R4B0: monochromatic red light).ResultsOur results showed that light quality significantly drove morphology, biomass accumulation, and flavonoids biosynthesis in S. baicalensis. R0B4 treatment promoted growth and flavonoids accumulation, including baicalin, and wogonoside concentrations. In contrast, UV-A radiation and green light negatively impacted these parameters compared to CK treatment. Interestingly, plant biomass and flavonoid concentrations were lower in R1B3, R1B1 and R3B1 treatments compared to monochromatic blue or red light.DiscussionOur study found that red light may antagonize blue light-stimulated growth and flavonoids accumulation, indicating a complex crosstalk between photoreceptors. These findings highlight the importance of blue light for optimizing the yield and quality of S. baicalensis in the understorey cultivation. It provides practice suggestion for the efficient management and sustainable cultivation of understorey medicinal plants.

Abstract Background Pruning is an important cultivation management option that has important effects on peach yield and quality. However, the effects of pruning on the overall genetic and metabolic changes in peach leaves and fruits are poorly understood. Results The transcriptomic and metabolomic profiles of leaves and fruits from trees subjected to pruning and unpruning treatments were measured. A total of 20,633 genes and 622 metabolites were detected. Compared with those in the control, 1,127 differentially expressed genes (DEGs) and 77 differentially expressed metabolites (DEMs) were identified in leaves from pruned and unpruned trees (pdLvsupdL), whereas 423 DEGs and 29 DEMs were identified in fruits from the pairwise comparison pdFvsupdF. The content of three auxin analogues was upregulated in the leaves of pruned trees, the content of all flavonoids detected in the leaves decreased, and the expression of almost all genes involved in the flavonoid biosynthesis pathway decreased. The phenolic acid and amino acid metabolites detected in fruits from pruned trees were downregulated, and all terpenoids were upregulated. The correlation analysis revealed that DEGs and DEMs in leaves were enriched in tryptophan metabolism, auxin signal transduction, and flavonoid biosynthesis. DEGs and DEMs in fruits were enriched in flavonoid and phenylpropanoid biosynthesis, as well as L-glutamic acid biosynthesis. Conclusions Pruning has different effects on the leaves and fruits of peach trees, affecting mainly the secondary metabolism and hormone signalling pathways in leaves and amino acid biosynthesis in fruits.

Abstract Background Anthocyanins are important contributors to coloration across a wide phylogenetic range of plants. Biological functions of anthocyanins span from reproduction to protection against biotic and abiotic stressors. Owing to a clearly visible phenotype of mutants, the anthocyanin biosynthesis and its sophisticated regulation have been studied in numerous plant species. Genes encoding the anthocyanin biosynthesis enzymes are regulated by a transcription factor complex comprising MYB, bHLH and WD40 proteins. Results A systematic comparison of anthocyanin-pigmented vs. non-pigmented varieties was performed within numerous plant species covering the taxonomic diversity of flowering plants. The literature was screened for cases in which genetic factors causing anthocyanin loss were reported. Additionally, transcriptomic data sets from four previous studies were reanalyzed to determine the genes possibly responsible for color variation based on their expression pattern. The contribution of different structural and regulatory genes to the intraspecific pigmentation differences was quantified. Differences concerning transcription factors are by far the most frequent explanation for pigmentation differences observed between two varieties of the same species. Among the transcription factors in the analyzed cases, MYB genes are significantly more prone to account for pigmentation differences compared to bHLH or WD40 genes. Among the structural genes, DFR genes are most often associated with anthocyanin loss. Conclusions These findings support previous assumptions about the susceptibility of transcriptional regulation to evolutionary changes and its importance for the evolution of novel coloration phenotypes. Our findings underline the particular significance of MYBs and their apparent prevalent role in the specificity of the MBW complex.

Abstract Background Pb stress, a toxic abiotic stress, critically affects maize production and food security. Although some progress has been made in understanding the damage caused by Pb stress and plant response strategies, the regulatory mechanisms and resistance genes involved in the response to lead stress in crops are largely unknown. Results In this study, to uncover the response mechanism of maize to Pb stress phenotype, physiological and biochemical indexes, the transcriptome, and the metabolome under different concentrations of Pb stress were combined for comprehensive analysis. As a result, the development of seedlings and antioxidant system were significantly inhibited under Pb stress, especially under relatively high Pb concentrations. Transcriptome analysis revealed 3559 co-differentially expressed genes(co-DEG) under the four Pb concentration treatments (500 mg/L, 1000 mg/L, 2000 mg/L, and 3000 mg/L Pb(NO3)2), which were enriched mainly in the GO terms related to DNA-binding transcription factor activity, response to stress, response to reactive oxygen species, cell death, the plasma membrane and root epidermal cell differentiation. Metabolome analysis revealed 72 and 107 differentially expressed metabolites (DEMs) under T500 and T2000, respectively, and 36 co-DEMs. KEGG analysis of the DEMs and DEGs revealed a common metabolic pathway, namely, flavonoid biosynthesis. An association study between the flavonoid biosynthesis-related DEMs and DEGs revealed 20 genes associated with flavonoid-related metabolites, including 3 for genistin and 17 for calycosin. Conclusion In summary, the study reveals that flavonoid metabolism plays an important role in response to Pb stress in maize, which not only provides genetic resources for the genetic improvement of maize Pb tolerance in the future but also enriches the theoretical basis of the maize Pb stress response.

This research investigates the flavonoid biosynthesis pathways of two ecotypes of Leymus chinensis, distinguished by their gray-green (GG) and yellow-green (YG) leaf colors, to uncover the molecular bases of their adaptability to different environmental conditions. By integrating comprehensive transcriptomic and metabolomic analyses, we identified 338 metabolites, with 161 showing differential expression—124 upregulated and 37 downregulated. The transcriptomic data revealed substantial variation, with 50,065 genes differentially expressed between the ecotypes, suggesting complex genetic regulation of the flavonoid biosynthesis pathways involving 20 enzyme-coding genes. KEGG pathway enrichment analysis further highlighted the involvement of 26 genes in the synthesis of four distinct types of flavonoid metabolites, indicating the sophisticated modulation of these pathways. Our results demonstrate that the GG and YG ecotypes of Leymus chinensis exhibit distinct flavonoid profiles and gene expression patterns, with the GG ecotype showing a higher accumulation of quercetin and kaempferol (increased by 25% and 33%, respectively, compared to YG), suggesting enhanced antioxidant capacity. Conversely, the YG ecotype displayed a broader spectrum of flavonoid metabolites, possibly indicating an adaptive strategy favoring diverse ecological interactions. Our results show that the GG and YG ecotypes of Leymus chinensis exhibit distinct flavonoid profiles and gene expression patterns, suggesting divergent adaptive strategies to environmental stress. This study highlights the crucial role of flavonoid metabolites in plant adaptation strategies, enhancing our understanding of plant resilience and adaptability. The distinct metabolic profiles observed suggest that the GG ecotype may be better equipped to handle oxidative stress, while the YG ecotype could be predisposed to broader ecological interactions. This emphasizes the value of applying machine learning in predicting plant adaptability, providing a new perspective for the future exploration of how plants adapt to environmental challenges. Meanwhile, the information gleaned from this nuanced study offers a foundation for future investigations into the genetic and environmental factors involved in plant adaptation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Pb stress, a toxic abiotic stress, critically affects maize production and food security. Although some progress has been made in understanding the damage caused by Pb stress and plant response strategies, the regulatory mechanisms and resistance genes involved in the response to lead stress in crops are largely unknown. Results: In this study, to uncover the response mechanism of maize to Pb stress phenotype, physiological and biochemical indexes, the transcriptome, and the metabolome under different concentrations of Pb stress were combined for comprehensive analysis. As a result, the development of seedlings and antioxidant system were significantly inhibited under Pb stress, especially under relatively high Pb concentrations. Transcriptome analysis revealed 3559 co-differentially expressed genes(co-DEG) under the four Pb concentration treatments (500 mg/L, 1000 mg/L, 2000 mg/L, and 3000 mg/L Pb(NO3)2), which were enriched mainly in the GO terms related to DNA-binding transcription factor activity, response to stress, response to reactive oxygen species, cell death, the plasma membrane and root epidermal cell differentiation. Metabolome analysis revealed 72 and 107 differentially expressed metabolites (DEMs) under T500 and T2000, respectively, and 36 co-DEMs. KEGG analysis of the DEMs and DEGs revealed a common metabolic pathway, namely, flavonoid biosynthesis. An association study between the flavonoid biosynthesis-related DEMs and DEGs revealed 20 genes associated with flavonoid-related metabolites, including 3 for genistin and 17 for calycosin. Conclusion: In summary, the study reveals that flavonoid metabolism plays an important role in response to Pb stress in maize, which not only provides genetic resources for the genetic improvement of maize Pb tolerance in the future but also enriches the theoretical basis of the maize Pb stress response. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Polygonati rhizoma, known for its distinct yellow rhizomes, is a common therapeutic and culinary plant in Far East Asia. The hue of medicinal plants is closely tied to the flavonoid biosynthesis and content levels. In this research, the fibrous root and taproot of Polygonatum kingianum Coll.et Hemsl. were studied to explore the secondary metabolite expression and flavonoid biosynthesis mechanisms using transcriptomics and metabolomics. Metabolic analysis identified that the differentially accumulated metabolites (DAMs) in the fibrous root and taproot were predominantly flavonoids, steroids, alkaloids, and phenolic acids. Overall, 200 flavonoids were identified in P. kingianum Coll.et Hemsl., with 170 exhibiting variances between the fibrous root and taproot. The transcriptome analysis revealed that a total of 289 unigenes encoding 32 enzymes were annotated into four flavonoid biosynthesis pathways, which include phenylpropanoid biosynthesis pathway, flavonoid biosynthesis pathway, isoflavonoid biosynthesis pathway, and flavone and flavonol biosynthesis pathway. The integration of transcriptomic and metabolomic data elucidated that the 76 differentially expressed genes (DEGs) encoding 13 enzyme genes (HCT, CCOMT, C4H, C3′H, CHI, PGT1, FLS, F3′H, CHS, ANR, DFR, F3′5′H, and LAR) and 15 DAMs preferred to be regulated in the flavonoid biosynthesis pathway. The expression of 10 DEGs was validated by qRT-PCR, agreeing with the same results by RNA-Seq. These findings shed light into the biosynthesis of secondary metabolites in P. kingianum Coll.et Hemsl., offering valuable information for the sustainable utilization and enhancement of this plant species. [ABSTRACT FROM AUTHOR]

Abiotic stress, especially drought stress, poses a significant threat to terrestrial plant growth, development, and productivity. Although mulberry has great genetic diversity and extensive stress-tolerant traits in agroforestry systems, only a few reports offer preliminary insight into the biochemical responses of mulberry leaves under drought conditions. In this study, we performed a comparative metabolomic and transcriptomic analysis on the "drooping mulberry" (Morus alba var. pendula Dippel) under PEG-6000-simulated drought stress. Our research revealed that drought stress significantly enhanced flavonoid accumulation and upregulated the expression of phenylpropanoid biosynthetic genes. Furthermore, the activities of superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) content were elevated. In vitro enzyme assays and fermentation tests indicated the involvement of flavonol synthase/flavanone 3-hydroxylase (XM_010098126.2) and anthocyanidin 3-O-glucosyltransferase 5 (XM_010101521.2) in the biosynthesis of flavonol aglycones and glycosides, respectively. The recombinant MaF3GT5 protein was found to recognize kaempferol, quercetin, and UDP-glucose as substrates but not 3-/7-O-glucosylated flavonols and UDP-rhamnose. MaF3GT5 is capable of forming 3-O- and 7-O-monoglucoside, but not di-O-glucosides, from kaempferol. This implies its role as a flavonol 3, 7-O-glucosyltransferase. The findings from this study provided insights into the biosynthesis of flavonoids and could have substantial implications for the future diversified utilization of mulberry. [ABSTRACT FROM AUTHOR]

Background: Pruning is an important cultivation management option that has important effects on peach yield and quality. However, the effects of pruning on the overall genetic and metabolic changes in peach leaves and fruits are poorly understood. Results: The transcriptomic and metabolomic profiles of leaves and fruits from trees subjected to pruning and unpruning treatments were measured. A total of 20,633 genes and 622 metabolites were detected. Compared with those in the control, 1,127 differentially expressed genes (DEGs) and 77 differentially expressed metabolites (DEMs) were identified in leaves from pruned and unpruned trees (pdLvsupdL), whereas 423 DEGs and 29 DEMs were identified in fruits from the pairwise comparison pdFvsupdF. The content of three auxin analogues was upregulated in the leaves of pruned trees, the content of all flavonoids detected in the leaves decreased, and the expression of almost all genes involved in the flavonoid biosynthesis pathway decreased. The phenolic acid and amino acid metabolites detected in fruits from pruned trees were downregulated, and all terpenoids were upregulated. The correlation analysis revealed that DEGs and DEMs in leaves were enriched in tryptophan metabolism, auxin signal transduction, and flavonoid biosynthesis. DEGs and DEMs in fruits were enriched in flavonoid and phenylpropanoid biosynthesis, as well as L-glutamic acid biosynthesis. Conclusions: Pruning has different effects on the leaves and fruits of peach trees, affecting mainly the secondary metabolism and hormone signalling pathways in leaves and amino acid biosynthesis in fruits. [ABSTRACT FROM AUTHOR]

Background: Breeding programs for nutrient-efficient tea plant varieties could be advanced by the combination of genotyping and phenotyping technologies. This study was aimed to search functional SNPs in key genes related to the nitrogen-assimilation in the collection of tea plant Camellia sinensis (L.) Kuntze. In addition, the objective of this study was to reveal efficient vegetation indices for phenotyping of nitrogen deficiency response in tea collection. Methods: The study was conducted on the tea plant collection of Camellia sinensis (L.) Kuntze of Western Caucasus grown without nitrogen fertilizers. Phenotypic data was collected by measuring the spectral reflectance of leaves in the 350–1100 nm range calculated as vegetation indices by the portable hyperspectral spectrometer Ci710s. Single nucleotide polymorphisms were identified in 30 key genes related to nitrogen assimilation and tea quality. For this, pooled amplicon sequencing, SNPs annotation and effect prediction with SnpEFF tool were used. Further, a linear regression model was applied to reveal associations between the functional SNPs and the efficient vegetation indices. Results: PCA and regression analysis revealed significant vegetation indices with high R2 values (more than 0.5) and the most reliable indices to select ND-tolerant genotypes were established: ZMI, CNDVI, RENDVI, VREI1, GM2, GM1, PRI, and Ctr2, VREI3, VREI2. The largest SNPs frequency was observed in several genes, namely F3'5'Hb, UFGTa, UFGTb, 4Cl, and AMT1.2. SNPs in NRT2.4, PIP, AlaDC, DFRa, and GS1.2 were inherent in ND-susceptible genotypes. Additionally, SNPs in AlaAT1, MYB4, and WRKY57, were led to alterations in protein structure and were observed in ND-susceptible tea genotypes. Associations were revealed between flavanol reflectance index (FRI) and SNPs in ASNb and PIP, that change the amino acids. In addition, two SNPs in 4Cl were associated with water band index (WBI). Conclusions: The results will be useful to identify tolerant and susceptible tea genotypes under nitrogen deficiency. Revealed missense SNPs and associations with vegetation indices improve our understanding of nitrogen effect on tea quality. The findings in our study would provide new insights into the genetic basis of tea quality variation under the N-deficiency and facilitate the identification of elite genes to enhance tea quality. [ABSTRACT FROM AUTHOR]

Background: Anthocyanins are important contributors to coloration across a wide phylogenetic range of plants. Biological functions of anthocyanins span from reproduction to protection against biotic and abiotic stressors. Owing to a clearly visible phenotype of mutants, the anthocyanin biosynthesis and its sophisticated regulation have been studied in numerous plant species. Genes encoding the anthocyanin biosynthesis enzymes are regulated by a transcription factor complex comprising MYB, bHLH and WD40 proteins. Results: A systematic comparison of anthocyanin-pigmented vs. non-pigmented varieties was performed within numerous plant species covering the taxonomic diversity of flowering plants. The literature was screened for cases in which genetic factors causing anthocyanin loss were reported. Additionally, transcriptomic data sets from four previous studies were reanalyzed to determine the genes possibly responsible for color variation based on their expression pattern. The contribution of different structural and regulatory genes to the intraspecific pigmentation differences was quantified. Differences concerning transcription factors are by far the most frequent explanation for pigmentation differences observed between two varieties of the same species. Among the transcription factors in the analyzed cases, MYB genes are significantly more prone to account for pigmentation differences compared to bHLH or WD40 genes. Among the structural genes, DFR genes are most often associated with anthocyanin loss. Conclusions: These findings support previous assumptions about the susceptibility of transcriptional regulation to evolutionary changes and its importance for the evolution of novel coloration phenotypes. Our findings underline the particular significance of MYBs and their apparent prevalent role in the specificity of the MBW complex. [ABSTRACT FROM AUTHOR]

Epimedium sagittatum (Sieb. et Zucc.) Maxim. (ESM) which accumulates several principal flavonoid compounds including epimedin A, B, C and icariin, is extensively utilized in traditional herbs for sexual dysfunction, osteoporosis etc. In China, ESM has a wealth of wild plant resources and characterized by significant variability in medicinal compounds accumulation. Understanding the diversity of ESMs can lead to better utilization of these plant resources. In this study, we integrated the metabolomic and transcriptomic analysis of three ESMs that originated in Anhui, Hubei and Jiangxi in China. Results showed that the flavonoid biosynthesis as well as the related gene expression in these ESMs revealed substantial differences. For example, the epimedin A, B, C and icariin as well as some related gene expression in ESMs from Anhui are significantly lower than those of in others. These results suggested that the ESMs from wild population without quality checkout may not be suitable for directly use as the materials for preparation of Chinese medicine and ESMs with different accumulation of metabolites could be used for distinct applications. [ABSTRACT FROM AUTHOR]

In nature, plants have developed a series of resistance mechanisms to face various external stresses. As understanding of the molecular mechanisms underlying plant resistance continues to deepen, exploring endogenous resistance in plants has become a hot topic in this field. Despite the multitude of studies on plant-induced resistance, how plants respond to stress under natural conditions remains relatively unclear. To address this gap, we investigated Chinese pine (Pinus tabuliformis) using pine caterpillar (Dendrolimus tabulaeformis) under natural conditions. Healthy Chinese pine trees, approximately 10 years old, were selected for studying induced resistance in Huangtuliangzi Forestry, Pingquan City, Chengde City, Hebei Province, China. Pine needles were collected at 2 h and 8 h after feeding stimulation (FS) via 10 pine caterpillars and leaf clipping control (LCC), to simulate mechanical damage caused by insect chewing for the quantification of plant hormones and transcriptome and metabolome assays. The results show that the different modes of treatments significantly influence the contents of JA and SA in time following treatment. Three types of differentially accumulated metabolites (DAMs) were found to be involved in the initial response, namely phenolic acids, lipids, and flavonoids. Weighted gene co-expression network analysis indicated that 722 differentially expressed genes (DEGs) are positively related to feeding stimulation and the specific enriched pathways are plant hormone signal transduction and flavonoid biosynthesis, among others. Two TIFY transcription factors (PtTIFY54 and PtTIFY22) and a MYB transcription factor (PtMYB26) were found to be involved in the interaction between plant hormones, mainly in the context of JA signal transduction and flavonoid biosynthesis. The results of this study provide an insight into how JA activates, serving as a reference for understanding the molecular mechanisms of resistance formation in conifers responding to mandibulate insects. [ABSTRACT FROM AUTHOR]

Polygonatum cyrtonema Hua (P. cyrtonema) rhizomes are rich in flavonoids and other secondary metabolites, exhibiting remarkable antioxidant, anti-tumor, and immunomodulatory effects. Polygonatum flavonoid-biosynthesis-related genes have been characterized already. However, a comprehensive overview of Polygonatum flavonoid biosynthesis pathways is still absent. To articulate the accumulation of the flavonoid biosynthesis pathways, we examined transcriptome changes using Illumina HiSeq from five different tissues and the RNA-seq of 15 samples had over 105 Gb of a clean base, generating a total of 277,955 unigenes. The cDNA libraries of the fruits (F), leaves (L), roots (R), stems (S), and rhizomes (T) of three-year-old P. cyrtonema plants generated 57,591, 53,578, 60,321, 51,530, and 54,935 unigenes. Comparative transcriptome analysis revealed that 379 differentially expressed genes (DEGs) were in the group of F _vs_ T, L _vs_ T, R _vs_ T, and S _vs_ T, and the transcripts of flavonoid-biosynthesis-related DEGs were principally enriched in rhizomes. In addition, combined with WGCNA and the FPKM of five tissues' transcription, nine differentially expressed transcription factor families (MYB, WRKY, AP2/ERF, etc.) were characterized in the red module, the red module positively correlated with rhizome flavonoid accumulation. Quantitative real-time PCR (qRT-PCR) further indicated that BZIP1, C3H31, ERF114, and DREB21 are differentially expressed in rhizomes, accompanied in rhizome development in P. cyrtonema. Therefore, this study provides a foundation for further research into uncovering the accumulation of flavonoid biosynthesis in the rhizomes of P. cyrtonema. [ABSTRACT FROM AUTHOR]