Synthesis Of Flavonoids Research Articles

Chromosome-scale genome analysis of Apocynum venetum sheds light on Apocynum phylogenetics, bast fiber development, and flavonoid synthesis

Apocynum venetum is a natural bast fiber crop and officinal plant belonging to the family Apocynaceae. Here, a chromosome-scale genome sequence was assembled using the single molecule sequencing and Hi-C technology. The genome of A. venetum is 228.80 Mb in length and has an N50 of 3.43 Mb. Up to 99.82 % of the contigs were attached to 11 pseudo-chromosomes. A total of 21,327 protein-coding genes were annotated in A. venetum genome. Phylogenetic analysis revealed the evolutionary relationships among the genera of Apocynum, Calotropis, and Catharanthus. Genes involved in bast fiber synthesis were identified in A. venetum by comparing gene expression between stem barks in different locations on the main stem. Combined metabolomic and transcriptomic analysis revealed the coordinated changes in flavonoids and related genes under methyl jasmonate treatment, as well as the dynamic change patterns of the genes encoding enzymes in the biosynthesis of flavonoids. Co-expression network analysis identified key genes that regulate flavonoid biosynthesis, including CHI (Av02G001490 and Av02G016020), MYB (Av01G029390), and bHLH (Av08G002930). The genome assembly of A. venetum provides a useful data set for understanding the evolution of Apocynum, bast fiber synthesis, and flavonoid biosynthesis in A. venetum.

Epigallocatechin-3-gallate-induced tolerance to cadmium stress involves increased flavonoid synthesis and nutrient homeostasis in tomato roots

Cadmium (Cd) is a toxic heavy metal, increasingly accumulating in the environment and its presence in various environmental compartments represents a significant risk to human health via the food chain. Epigallocatechin-3-Gallate (EGCG) is a prominent secondary metabolite, which can safeguard plants from biotic and abiotic stress. However, the role of EGCG in flavonoid synthesis, nutrient acquisition and reactive oxygen species (ROS) metabolism under Cd stress remains unclear. Here, we examined the effects of EGCG and Cd treatment on leaf photochemical efficiency, cell ultrastructure, essential element acquisition, antioxidant system, and secondary metabolism in tomato (Solanum lycopersicum L.). The results showed that O2•−, H2O2, and malondialdehyde levels increased after Cd treatment, but Fv/Fm decreased significantly, suggesting that Cd induced oxidative stress and photoinhibition. However, EGCG mitigated the adverse effects of Cd-induced phytotoxicity in both the roots and leaves. A decrease in ROS accumulation under EGCG + Cd treatment was mainly attributed to the significant enhancement in antioxidant enzyme activity, flavonoid content, and PHENYLALANINE AMMONIA-LYASE expression in roots. Moreover, EGCG reduced Cd content but increased some essential nutrient contents in tomato plants. Transmission electron microscopy-based observations revealed that EGCG treatment safeguards leaf and root cell ultrastructure under Cd stress. This implies that tomato plants subjected to Cd stress experienced advantageous effects upon receiving EGCG treatment. The present work elucidated critical mechanisms by which EGCG induces tolerance to Cd, thereby providing a basis for future investigations into environmentally sustainable agricultural practices in areas contaminated with heavy metals, for utilizing naturally occurring substances found in plants.

Analysis of metabolic differences between Jiaosu fermented from dendrobium flowers and stems based on untargeted metabolomics

Dendrobium officinale is an important traditional Chinese medicinal herb containing bioactive polysaccharides and alkaloids. This study characterized metabolite differences between jiaosu (fermented plant product) from Dendrobium flowers versus stems using untargeted metabolomics. The jiaosu was fermented by mixed fermentation of Saccharomyces cerevisiae, Lactobacillus bulgaricus and Streptococcus thermophilus. Liquid chromatography-mass spectrometry analysis identified 476 differentially expressed metabolites between the two Jiaosu products. Key results showed downregulation of flavonoid metabolism in Dendrobium Stems Edible Plant Jiaosu (SEP) but increased flavonoid synthesis in Dendrobium Flowers Edible Plant Jiaosu (FEP), likely an antioxidant response. SEP displayed upregulation of lignin metabolites with potential antioxidant properties. The findings demonstrate significant metabolite profile differences between SEP and FEP, providing the basis for developing functional jiaosu products targeting specific health benefits.

Multi-omics profiling reveal responses of three major Dendrobium species from different growth years to medicinal components.

Dendrobium is a perennial herb found in Asia that is known for its medicinal and ornamental properties. Studies have shown that the stem is the primary medicinal component of Dendrobium spp. To investigate the effect of the species and age of Dendrobium (in years) on the content of its medicinal components, we collected the stems of 1-to-4-year-old D. officinale, D. moniliforme, and D. huoshanense, sequenced the transcriptome, metabolome, and microbiome, and analyzed the data in a comprehensive multi-omics study. We identified 10,426 differentially expressed genes (DEGs) with 644 differentially accumulated metabolites (DAMs) from 12 comparative groups and mapped the flavonoid pathway based on DEGs and DAMs. Transcriptomic and metabolomic data indicated a general trend of the accumulation of flavonoids exhibiting pharmacological effects in the three Dendrobium species. In addition, joint metabolome and microbiome analyses showed that actinobacteria was closely associated with flavonoid synthesis with increasing age. Our findings provide novel insights into the interactions of flavonoids of Dendrobium with the transcriptome and microbiome.

Transcriptomic and targeted metabolome analyses revealed the regulatory mechanisms of the synthesis of bioactive compounds in Citrus grandis 'tomentosa'.

Exocarpium Citri Grandis is a popular Chinese herbal medicine prepared from Citrus grandis 'tomentosa', and it is rich in several bioactive compounds, including flavonoids, coumarins, and volatile oils. However, studies are yet to elucidate the mechanisms of synthesis and regulation of these active components. Therefore, the present study examined the profiles of flavonoids and volatile oil bioactive compounds in plant petals, fruits, and tender leaves, and then performed RNA sequencing on different tissues to identify putative genes involved in the synthesis of bioactive compounds. The results show that the naringin, naringenin, and coumarin contents of the fruitlets were significantly higher than those of the tender leaves and petals, whereas the tender leaves had significantly higher levels of rhoifolin and apigenin. A total of 49 volatile oils, of which 10 were mainly found in flowers, 15 were mainly found in fruits, and 18 were mainly found in leaves, were identified. RNA sequencing identified 9,942 genes that were differentially expressed in different tissues. Further analysis showed that 20, 15, and 74 differentially expressed genes were involved in regulating flavonoid synthesis, regulating coumarin synthesis, and synthesis and regulation of terpenoids, respectively. CHI1 (Cg7g005600) and 1,2Rhat gene (Cg1g023820) may be involved in the regulation of naringin synthesis in C. grandis fruits. The HDR (Cg8g006150) gene, HMGS gene (Cg5g009630) and GGPS (Cg1g003650) may be involved in the regulation and synthesis of volatile oils in C. grandis petals. Overall, the findings of the present study enhance our understanding of the regulatory mechanisms of secondary metabolites in C. grandis, which could promote the breeding of C. grandis with desired characteristics.

Major quality regulation network of flavonoid synthesis governing the bioactivity of black wolfberry.

Black wolfberry (Lycium ruthenicum Murr.) contains various bioactive metabolites represented by flavonoids, which are quite different among production regions. However, the underlying regulation mechanism of flavonoid biosynthesis governing the bioactivity of black wolfberry remains unclear. Presently, we compared the bioactivity of black wolfberry from five production regions. Multi-omics were performed to construct the regulation network associated with the fruit bioactivity. The detailed regulation mechanisms were identified using genetic and molecular methods. Typically, Qinghai (QH) fruit exhibited higher antioxidant and anti-inflammatory activities. The higher medicinal activity of QH fruit was closely associated with the accumulation of eight flavonoids, especially Kaempferol-3-O-rutinoside (K3R) and Quercetin-3-O-rutinoside (rutin). Flavonoid biosynthesis was found to be more active in QH fruit, and the upregulation of LrFLS, LrCHS, LrF3H and LrCYP75B1 caused the accumulation of K3R and rutin, leading to high medicinal bioactivities of black wolfberry. Importantly, transcription factor LrMYB94 was found to regulate LrFLS, LrCHS and LrF3H, while LrWRKY32 directly triggered LrCYP75B1 expression. Moreover, LrMYB94 interacted with LrWRKY32 to promote LrWRKY32-regulated LrCYP75B1 expression and rutin synthesis in black wolfberry. Transgenic black wolfberry overexpressing LrMYB94/LrWRKY32 contained higher levels of K3R and rutin, and exhibited high medicinal bioactivities. Importantly, the LrMYB94/LrWRKY32-regulated flavonoid biosynthesis was light-responsive, showing the importance of light intensity for the medicinal quality of black wolfberry. Overall, our results elucidated the regulation mechanisms of K3R and rutin synthesis, providing the basis for the genetic breeding of high-quality black wolfberry.

A novel transcription factor FnMYB4 regulates pigments metabolism of yellow leaf mutants in Fragaria nilgerrensis

The strawberry species Fragaria nilgerrensis Schlechtendal ex J. Gay, renowned for its distinctive white, fragrant peach-like fruits and strong disease resistance, is an exceptional research material. In a previous study, an ethyl methane sulfonate (EMS) mutant library was established for this species, resulting in various yellow leaf mutants. Leaf yellowing materials are not only the ideal materials for basic studies on photosynthesis mechanism, chloroplast development, and molecular regulation of various pigments, but also have important utilization value in ornamental plants breeding. The present study focused on four distinct yellow leaf mutants: mottled yellow leaf (MO), yellow green leaf (YG), light green leaf (LG), and buddha light leaf (BU). The results revealed that the flavonoid content and carotenoid-to-chlorophyll ratio exhibited a significant increase among these mutants, while experiencing a significant decrease in chlorophyll and carotenoid contents compared to the wild type (WT). To clarify the regulatory mechanisms and network relationships underlying these mutants, the RNA-seq and weighted gene co-expression network (WGCNA) analyses were employed. The results showed flavonoid metabolism pathway was enriched both in MO and YG mutants, while the chlorophyll biosynthesis pathway and carotenoid degradation pathway were only enriched in MO and YG mutants, respectively. Subsequently, key structural genes and transcription factors were identified on metabolic pathways of three pigments through correlation analyses and quantitative experiments. Furthermore, a R2R3-MYB transcription factor, FnMYB4, was confirmed to be positively correlated with flavonoid synthesis through transient overexpression, virus-induced gene silencing (VIGS), and RNA interference (RNAi), accompanying by reoccurrence and attenuation of mutant phenotype. Finally, dual-luciferase (LUC) and yeast one-hybrid assays confirmed the binding of FnMYB4 to the FnFLS and FnF3H promoters, indicating that FnMYB4 positively regulates flavonoid synthesis. In addition, correlation analyses suggested that FnMYB4 also might be involved in chlorophyll and carotenoid metabolisms. These findings demonstrated the pivotal regulatory role of FnMYB4 in strawberry leaf coloration.

Integrated transcriptomic and metabolomic analyses elucidate the mechanism of flavonoid biosynthesis in the regulation of mulberry seed germination under salt stress

Seed propagation is the main method of mulberry expansion in China, an important economic forest species. However, seed germination is the most sensitive stage to various abiotic stresses, especially salinity stress. To reveal the molecular regulatory mechanism of mulberry seed germination under salt stress, flavonoid metabolomics and transcriptomics analyses were performed on mulberry seeds germinated under 50 and 100 mmol/L NaCl stress. Analysis of the flavonoid metabolome revealed that a total of 145 differential flavonoid metabolites (DFMs) were classified into 9 groups, 40 flavonols, 32 flavones, 16 chalcones and 14 flavanones. Among them, 61.4% (89) of the DFMs accumulated continuously with increasing salt concentration, reaching the highest level at a 100 mmol/L salt concentration; these DFMs included quercetin-3-O-glucoside (isoquercitrin), kaempferol (3,5,7,4'-tetrahydroxyflavone), quercetin-7-O-glucoside, taxifolin (dihydroquercetin) and apigenin (4',5,7-trihydroxyflavone), indicating that these flavonoids may be key metabolites involved in the response to salt stress. Transcriptional analysis identified a total of 3055 differentially expressed genes (DEGs), most of which were enriched in flavonoid biosynthesis (ko00941), phenylpropanoid biosynthesis (ko00940) and biosynthesis of secondary metabolites (ko01110). Combined analysis of flavonoid metabolomic and transcriptomic data indicated that phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), flavonol synthase (FLS), bifunctional dihydroflavonol 4-reductase/flavanone 4-reductase (DFR) and anthocyanidin reductase (ANR) were the key genes involved in flavonoid accumulation during mulberry seed germination under 50 and 100 mmol/L NaCl stress. In addition, three transcription factors, MYB, bHLH and NAC, were involved in the regulation of flavonoid accumulation under salt stress. The results of quantitative real-time PCR (qRT‒PCR) validation showed that the expression levels of 11 DEGs, including 7 genes involved in flavonoid biosynthesis, under different salt concentrations were consistent with the transcriptomic data, and parallel reaction monitoring (PRM) results showed that the expression levels of 6 key enzymes (proteins) involved in flavonoid synthesis were consistent with the accumulation of flavonoids. This study provides a new perspective for investigating the regulatory role of flavonoid biosynthesis in the regulation of mulberry seed germination under salt stress at different concentrations.

Selenium Improves the Control Efficacy of Phytophthora nicotianae by Damaging the Cell Membrane System and Promoting Plant Energy Metabolism.

Tobacco black shank (TBS), caused by Phytophthora nicotianae, poses a significant threat to tobacco plants. Selenium (Se), recognized as a beneficial trace element for plant growth, exhibited inhibitory effects on P. nicotianae proliferation, disrupting the cell membrane integrity. This action reduced the energy supply and hindered hyphal transport through membrane proteins, ultimately inducing hyphal apoptosis. Application of 8 mg/L Se through leaf spraying resulted in a notable decrease in TBS incidence. Moreover, Se treatment preserved chloroplast structure, elevated chitinase activities, β-1,3-GA, polyphenol oxidase, phenylalanine ammonia-lyase, and increased hormonal content. Furthermore, Se enhanced flavonoid and sugar alcohol metabolite levels while diminishing amino acid and organic acid content. This shift promoted amino acid degradation and flavonoid synthesis. These findings underscore the potential efficacy of Se in safeguarding tobacco and potentially other plants against P. nicotianae.

Transcriptional alterations of peanut root during interaction with growth-promoting Tsukamurella tyrosinosolvens strain P9.

The plant growth-promoting rhizobacterium Tsukamurella tyrosinosolvens P9 can improve peanut growth. In this study, a co-culture system of strain P9 and peanut was established to analyze the transcriptome of peanut roots interacting with P9 for 24 and 72 h. During the early stage of co-culturing, genes related to mitogen-activated protein kinase (MAPK) and Ca2+ signal transduction, ethylene synthesis, and cell wall pectin degradation were induced, and the up-regulation of phenylpropanoid derivative, flavonoid, and isoflavone synthesis enhanced the defense response of peanut. The enhanced expression of genes associated with photosynthesis and carbon fixation, circadian rhythm regulation, indoleacetic acid (IAA) synthesis, and cytokinin decomposition promoted root growth and development. At the late stage of co-culturing, ethylene synthesis was reduced, whereas Ca2+ signal transduction, isoquinoline alkaloid synthesis, and ascorbate and aldarate metabolism were up-regulated, thereby maintaining root ROS homeostasis. Sugar decomposition and oxidative phosphorylation and nitrogen and fatty acid metabolism were induced, and peanut growth was significantly promoted. Finally, the gene expression of seedlings inoculated with strain P9 exhibited temporal differences. The results of our study, which explored transcriptional alterations of peanut root during interacting with P9, provide a basis for elucidating the growth-promoting mechanism of this bacterial strain in peanut.

Transcription factors FtMYB22 negatively regulates anthocyanin synthesis by forming an MBW ternary complex in Tartary buckwheat

AbstractAnthocyanins are important flavonoid compounds that are abundant in Tartary buckwheat, providing a variety of bright colors and enhancing abiotic stress resistance. Our study found that FtMYB22 specifically affected anthocyanin anabolism. In this study, overexpression of the FtMYB22 transcription factor in tobacco significantly reduced the accumulation of flavonol in the petals of transgenic tobacco plants. Moreover, it significantly downregulated the expression of the genes involved in anthocyanin biosynthesis in tobacco. Overexpression of the FtMYB22 transcription factor in Tartary buckwheat hairy roots significantly inhibited anthocyanin and total flavonoids biosynthesis, and downregulated the expression of the anthocyanin biosynthesis genes. Comparing wild‐type and transgenic hairy roots, FtMYB22 overexpression significantly inhibited anthocyanin accumulation and downregulated most flavonoid synthesis pathway gene expression. Yeast two‐hybrid assays demonstrated that FtMYB22 could interact with the FtbHLH family member FtTT8 and the FtWD40 family member FtTTG1 to form a ternary MBW complex to inhibit anthocyanin synthesis. Therefore, FtMYB22 can be exploited as a target site for further gene editing and molecular breeding to improve the resilience and quality of Tartary buckwheat.

Generation and characterisation of an Arabidopsis thaliana f3h/fls1/ans triple mutant that accumulates eriodictyol derivatives

BackgroundFlavonoids are plant specialised metabolites, which derive from phenylalanine and acetate metabolism. They possess a variety of beneficial characteristics for plants and humans. Several modification steps in the synthesis of tricyclic flavonoids cause for the amazing diversity of flavonoids in plants. The 2-oxoglutarate-dependent dioxygenases (2-ODDs) flavanone 3-hydroxylase (F3H, synonym FHT), flavonol synthase (FLS) and anthocyanidin synthase (ANS, synonym leucoanthocyanidin dioxygenase (LDOX)), catalyse oxidative modifications to the central C ring. They are highly similar and have been shown to catalyse, at least in part, each other’s reactions. FLS and ANS have been identified as bifunctional enzymes in many species, including Arabidopsis thaliana, stressing the capability of plants to bypass missing or mutated reaction steps on the way to flavonoid production. However, little is known about such bypass reactions and the flavonoid composition of plants lacking all three central flavonoid 2-ODDs.ResultsTo address this issue, we generated a f3h/fls1/ans mutant, as well as the corresponding double mutants and investigated the flavonoid composition of this mutant collection. The f3h/fls1/ans mutant was further characterised at the genomic level by analysis of a nanopore DNA sequencing generated genome sequence assembly and at the transcriptomic level by RNA-Seq analysis. The mutant collection established, including the novel double mutants f3h/fls1 and f3h/ans, was used to validate and analyse the multifunctionalities of F3H, FLS1, and ANS in planta. Metabolite analyses revealed the accumulation of eriodictyol and additional glycosylated derivatives in mutants carrying the f3h mutant allele, resulting from the conversion of naringenin to eriodictyol by flavonoid 3’-hydroxylase (F3’H) activity.ConclusionsWe describe the in planta multifunctionality of the three central flavonoid 2-ODDs from A. thaliana and identify a bypass in the f3h/fls1/ans triple mutant that leads to the formation of eriodictyol derivatives. As (homo-)eriodictyols are known as bitter taste maskers, the annotated eriodictyol (derivatives) and in particular the observations made on their in planta production, could provide valuable insights for the creation of novel food supplements.

Unraveling the Molecular Basis of Color Variation in Dioscorea alata Tubers: Integrated Transcriptome and Metabolomics Analysis.

Dioscorea alata L. (Dioscoreaceae) is a widely cultivated tuber crop with variations in tuber color, offering potential value as health-promoting foods. This study focused on the comparison of D. alata tubers possessing two distinct colors, white and purple, to explore the underlying mechanisms of color variation. Flavonoids, a group of polyphenols known to influence plant color and exhibit antioxidant properties, were of particular interest. The total phenol and total flavonoid analyses revealed that purple tubers (PTs) have a significantly higher content of these metabolites than white tubers (WTs) and a higher antioxidant activity than WTs, suggesting potential health benefits of PT D. alata. The transcriptome analysis identified 108 differentially expressed genes associated with the flavonoid synthesis pathway, with 57 genes up-regulated in PTs, including CHS, CHI, DFR, FLS, F3H, F3'5'H, LAR, ANS, and ANR. The metabolomics analysis demonstrated that 424 metabolites, including 104 flavonoids and 8 tannins, accumulated differentially in PTs and WTs. Notably, five of the top ten up-regulated metabolites were flavonoids, including 6-hydroxykaempferol-7-O-glucoside, pinocembrin-7-O-(6″-O-malonyl)glucoside, 6-hydroxykaempferol-3,7,6-O-triglycoside, 6-hydroxykaempferol-7-O-triglycoside, and cyanidin-3-O-(6″-O-feruloyl)sophoroside-5-O-glucoside, with the latter being a precursor to anthocyanin synthesis. Integrating transcriptome and metabolomics data revealed that the 57 genes regulated 20 metabolites within the flavonoid synthesis pathway, potentially influencing the tubers' color variation. The high polyphenol content and antioxidant activity of PTs indicate their suitability as nutritious and health-promoting food sources. Taken together, the findings of this study provide insights into the molecular basis of tuber color variation in D. alata and underscore the potential applications of purple tubers in the food industry and human health promotion. The findings contribute to the understanding of flavonoid biosynthesis and pigment accumulation in D. alata tubers, opening avenues for future research on enhancing the nutritional quality of D. alata cultivars.

Metabolic Patterns of Flavonoid and Its Key Gene Expression Characteristics of Five Cultivars of Tulipa gesneriana during Flower Development

Flower color is one of the most important ornamental traits of tulips (Tulipa gesneriana). Five typical tulip cultivars were selected to identify the flavonoid components and analyze their key gene expression in their tepals. Firstly, after preliminary determination of the pigment type, the flavonoids were identified by UPLC-Q-TOF-MS. A total of 17 anthoxanthins were detected in the five cultivars. The total anthoxanthin content in the white tulip and the red tulip showed a similar decreasing trend, while an increasing trend was observed in the black tulip. Similarly, a total of 13 anthocyanins were detected in five tulip cultivars. The black tulip contained the largest number of anthocyanins, mainly delphinidin derivatives (Dp) and cyanidin derivatives (Cy). The total anthocyanin content (TAC) in the orange, red, and black cultivars was higher than that in the white and yellow cultivars and presented an overall increase trend along with the flower development. TgCHS, TgFLS, TgF3H, TgF3′H, TgF3′5′H, and TgDFR, as key structural genes, were involved in the flavonoid synthesis pathway, and the expression patterns of these genes are basically consistent with the components and accumulation patterns of flavonoids mentioned above. Taken together, the flower color in tulips was closely related to the composition and content of anthocyanins and anthoxanthins, which were indeed regulated by certain key structural genes in the flavonoid pathway.

Integrated Transcriptomics and Metabolomics Analysis Reveals the Effects of Cutting on the Synthesis of Flavonoids and Saponins in Chinese Herbal Medicine Astragalus mongholious.

Astragali Radix, derived from the roots of Astragalus mongholicus, is a traditional Chinese medicine containing flavonoids and saponins as its key ingredients. With a shortage in the wild sources of the herbal plant, it is especially important to explore a cultivation mode for A. mongholicus for medicinal purposes. Cutting, a physical environmental stress method, was used in this study with the objective of improving the quality of this herbal legume. We found that cutting of the top 1/3 of the aboveground part of A. mongholicus during the fruiting period resulted in a significant increase in the content of flavonoids and saponins, as well as in root growth, including length, diameter, and dry weight. Furthermore, the leaves were sampled and analyzed using a combined transcriptome and metabolome analysis approach at five different time points after the treatment. Sixteen differentially expressed unigenes (DEGs) involved in the biosynthesis of flavonoids were identified; these were found to stimulate the synthesis of flavonoids such as formononetin and calycosin-7-O-β-D-glucoside. Moreover, we identified 10 DEGs that were associated with the biosynthesis of saponins, including astragaloside IV and soyasaponin I, and found that they only regulated the mevalonic acid (MVA) pathway. These findings provide new insights into cultivating high-quality A. mongholicus, which could potentially alleviate the scarcity of this valuable medicinal plant.

Changes in Biologically Active Compounds in Pinus sylvestris Needles after Lymantria monacha Outbreaks and Treatment with Foray 76B.

Due to climate warming, the occurrence of Lymantria monacha outbreaks is predicted to become more frequent, causing repeated and severe damage to conifer trees. Currently, the most effective way to control the outbreaks is aerial spraying with the bioinsecticide Foray 76B. The present study aimed to determine the impact of both: (i) L. monacha outbreaks and (ii) treatment with Foray 76B on tree resistance through the synthesis of polyphenols (TPC), flavonoids (TFC), photosynthetic pigments (chlorophyll a and b, carotenoids), lipid peroxidation (MDA), and soluble sugars (TSS) in Pinus sylvestris needles. Samples were collected from visually healthy (control), damaged/untreated, and damaged/Foray 76B-treated plots in 2020 and 2021 (following year after the outbreaks). The results revealed that L. monacha outbreaks contributed to the increase in TPC by 34.1% in 2020 and 26.7% in 2021. TFC negatively correlated with TPC, resulting in 17.6% and 11.1% lower concentrations in L. monacha-damaged plots in 2020 and 2021, respectively. A decrease in MDA was found in the damaged plots in both 2020 and 2021 (10.2% and 23.3%, respectively), which was associated with the increased synthesis of photosynthetic pigments in 2021. The research results also showed that in the following year after the outbreaks, the increase in the synthesis of photosynthetic pigments was also affected by the treatment with Foray 76B. Moreover, the increase in the synthesis of TPC and photosynthetic pigments in the damaged plots in 2021 illustrates the ability of pines to keep an activated defense system to fight biotic stress. Meanwhile, a higher synthesis of photosynthetic pigments in Foray 76B-treated plots indicates a possible effect of the treatment on faster tree growth and forest recovery after L. monacha outbreaks.

Effect of light intensity on celery growth and flavonoid synthesis.

Light is one of the important environmental factors affecting the growth and development of facility vegetables. In this experiment, we investigated the effects of different light intensities on the growth, nutritional quality and flavonoid accumulation of celery under hydroponic and full LED light conditions. Four light intensities of 40, 100, 200, or 300 µmol·m-2·s-1 were set up in the experiment, and three harvest periods were set up on the basis of different light intensities, which were 15, 30, and 45 d after treatment (labeled as S1, S2, and S3, respectively). The results showed that the plant height and aboveground biomass of celery increased with the increase of light intensity, and the light intensity of 200 μmol·m-2·s-1 was beneficial to increase the contents of chlorophyll, carotenoids, total phenols, vitamin C, cellulose, total flavones and apigenin in celery. During the S1-S3 period, the activities of PAL, CHS, CHI and ANS increased gradually under 200 and 300 μmol·m-2·s-1 light intensity treatments, and the activities of FNS and CHS enzymes were the highest under 200 μmol·m-2·s-1 light intensity treatment. The expression and ANS activity of Ag3GT, a key gene for anthocyanin synthesis, reached the maximum value at 300 μmol·m-2·s-1, and the expression level and FNS activity of AgFNS, a key gene for apigenin synthesis, reached a maximum value at 200 μmol·m-2·s-1. In general, the anthocyanin content was the highest at 300 μmol·m-2·s-1, and the apigenin content was the highest at 200 μmol·m-2·s-1. In conclusion, light intensity of 200 µmol·m-2·s-1 treatment was more favorable for celery growth and nutrient synthesis.

Integrated metabolome and transcriptome analysis reveals the regulatory mechanism of low nitrogen-driven biosynthesis of saponins and flavonoids in Panax notoginseng

BackgroundNitrogen (N) is an important macronutrient involved in the biosynthesis of primary and secondary metabolites in plants. However, the metabolic regulatory mechanism of low-N-induced triterpenoid saponin and flavonoid accumulation in rhizomatous medicinal Panax notoginseng (Burk.) F. H. Chen remains unclear. MethodsTo explore the potential regulatory mechanism and metabolic basis controlling the response of P. notoginseng to N deficiency, the transcriptome and metabolome were analysed in the roots. ResultsThe N content was significantly reduced in roots of N0-treated P. notoginseng (0 kg·N·667 m−2). The C/N ratio was enhanced in the N-deficient P. notoginseng. N deficiency promotes the accumulation of amino acids (L-proline, L-leucine, L-isoleucine, L-norleucine, L-arginine, and L-citrulline) and sugar (arabinose, xylose, glucose, fructose, and mannose), thus providing precursor metabolites for the biosynthesis of flavonoids and triterpenoid saponins. Downregulation of key structural genes (PAL, PAL3, ACC1, CHS2, PPO, CHI3, F3H, DFR, and FGT), in particular with the key genes of F3H, involved in the flavonoid biosynthesis pathway possibly induced the decrease in flavonoid content with increased N supply. Notoginsenoside R1, ginsenoside Re, Rg1, Rd, F1, R1 + Rg1 + Rb1 and total triterpenoid saponins were enhanced in the N0 groups than in the N15 (15 kg·N·667 m−2) plants. Higher phosphoenolpyruvate (an intermediate of glycolyticwith pathway metabolism) and serine (an intermediate of photorespiration) levels induced by N deficiency possibly promote saponin biosynthesis through mevalonic acid (MVA) and methylerythritol (MEP) pathways. Genes (MVD2, HMGS, HMGR1, HMGR2, DXR, and HMGR1) encoding the primary enzymes HMGS, HMGR, DXR, and MVD in the MVA and MEP pathways were significantly upregulated in the N0-treated P. notoginseng. The saponin biosynthesis genes DDS, DDS, CYP716A52, CYP716A47, UGT74AE2, and FPS were upregulated in the N-deficient plants. Upregulation of genes involved in saponin biosynthesis promotes the accumulation of triterpenoid saponins in the N0-grown P. notoginseng. ConclusionsN deficiency enhances primary metabolisms, such as amino acids and sugar accumulation, laying the foundation for the synthesis of flavonoids and triterpenoid saponins in P. notoginseng. F3H, DDS, FPS, HMGR, HMGS and UGT74AE2 can be considered as candidates for functional characterisation of the N-regulated accumulation of triterpenoid saponins and flavonoids in future.

Integrated metabolomic and transcriptomic analyses revealed metabolite variations and regulatory networks in Cinnamomum cassia Presl from four growth years.

To understand the mechanism of the dynamic accumulation of active ingredients in Cinnamomum cassia Presl, metabolomic and transcriptomic analyses of 5~8 years old C. cassia were performed. A total of 72 phenylpropanoids, 146 flavonoids, and 130 terpenoids showed marked changes. Most phenylpropanoids and flavonoids showed markedly higher abundances in 6-year-old C. cassia than in others, which was related to the higher expression of genes that synthesize and regulate phenylpropanoids and flavonoid. We identified transcription factors (TFs) and genes involved in phenylpropanoids and flavonoids synthesis and regulation through co-expression network analyses. Furthermore, most of the terpenoids in 5-year-old C. cassia showed markedly higher abundances than in others, which was due to the differentially expressed genes upstream of the terpenoids pathway. The results of our study provide new insights into the synthesis and accumulation of phenylpropanoid, flavonoids and terpenoids in C. cassia at four growth stages.

Transcriptome Analysis of Plant Growth-promoting Bacteria Alleviating Microplastic and Heavy Metal Combined Pollution Stress in Sorghum

Microplastics can become potential transport carriers of other environmental pollutants (such as heavy metals), so the combined pollution of microplastics and heavy metals has attracted increasing attention from researchers. To explore the mechanism of plant growth-promoting bacteria VY-1 alleviating the combined pollution stress of heavy metals and microplastics in sorghum, the effects of inoculation on biomass and accumulation of heavy metals in sorghum were analyzed using a hydroponics experiment, and the effects of inoculation on gene expression in sorghum were analyzed via transcriptomics. The results showed that the combined pollution of polyethylene (PE) and cadmium (Cd) decreased the dry weight of above-ground and underground parts by 17.04% and 10.36%, respectively, compared with that under the single Cd pollution, which showed that the combined toxicity effect of the combined pollution on plant growth was enhanced. The inoculation of plant growth-promoting bacteria VY-1 could alleviate the toxicity of Cd-PE combined pollution and increase the length of aboveground and underground parts by 33.83% and 73.21% and the dry weight by 56.64% and 33.44%, respectively. Transcriptome sequencing showed that 904 genes were up-regulated after inoculation with VY-1. Inoculation with growth-promoting bacteria VY-1 could up-regulate the expression of several genes in the auxin, abscisic acid, flavonoid synthesis, and lignin biosynthesis pathways, which promoted the response ability of sorghum under Cd-PE combined pollution stress and improved its resistance. The above results indicated that plant growth-promoting bacteria could alleviate the stress of heavy metal and microplastic combined pollution by regulating plant gene expression, which provided a reference for plant-microbial joint remediation of heavy metal and microplastic combined pollution.