Plant Secondary Metabolism Research Articles

Special Issue “Advances in the Physiology of Primary and Secondary Plant Metabolism Under Abiotic and Biotic Stress”

One of the most relevant areas of biology is the study of plant adaptation processes to the action of various stress factors of abiotic and biotic nature, which is reflected in the works of molecular biologists, geneticists, microbiologists, plant physiologists, and biochemists, as well as biotechnologists [1–10]. [...]

Transcriptome Analysis Reveals the Crucial Role of Phenylalanine Ammonia-Lyase in Low Temperature Response in Ammopiptanthus mongolicus

Background: Ammopiptanthus mongolicus is a rare temperate evergreen shrub with high tolerance to low temperature, and understanding the related gene expression regulatory network can help advance research on the mechanisms of plant tolerance to abiotic stress. Methods: Here, time-course transcriptome analysis was applied to investigate the gene expression network in A. mongolicus under low temperature stress. Results: A total of 12,606 differentially expressed genes (DEGs) were identified at four time-points during low temperature stress treatment, and multiple pathways, such as plant hormones, secondary metabolism, and cell membranes, were significantly enriched in the DEGs. Trend analysis found that the expression level of genes in cluster 19 continued to upregulate under low temperatures, and the genes in cluster 19 were significantly enriched in plant hormone signaling and secondary metabolic pathways. Based on the transcriptome data, the expression profiles of the genes in abscisic acid, salicylic acid, and flavonoid metabolic pathways were analyzed. It was found that biosynthesis of abscisic acid and flavonoids may play crucial roles in the response to low temperature stress. Furthermore, members of the phenylalanine ammonia-lyase (PAL) family in A. mongolicus were systematically identified and their structures and evolution were characterized. Analysis of cis-acting elements showed that the PAL genes in A. mongolicus were closely related to abiotic stress response. Expression pattern analysis showed that PAL genes responded to various environmental stresses, such as low temperature, supporting their involvement in the low temperature response in A. mongolicus. Conclusions: Our study provides important data for understanding the mechanisms of tolerance to low temperatures in A. mongolicus.

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

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

Antibacterial and Antibiofilm Activity of Essential Oils Against Aeromonas spp. Isolated from Rainbow Trout

Aeromonas spp. is a major pathogen in aquaculture with a great negative economic impact. Essential oils (EOs) are compounds of the secondary metabolism of plants known for their antibacterial and antibiofilm activities. In this study, in vitro antibacterial activity of eight EOs: tea tree (extracted from Melaleuca alternifolia), eucalyptus (extracted from Eucalyptus globulus LABILL.), knee timber (extracted from Pini mungo L.), peppermint (extracted from Mentha piperita L.), oregano (extracted from Origanum vulgare L.), rosemary (extracted from Rosmarinus officinalis L.), thyme (extracted from Thymus vulgaris L.) and pine EO (extracted from Pinus silvestris L.), obtained from Calendula a.s., was evaluated. Their antibacterial activity was demonstrated against Aeromonas spp. isolates. Oregano and thyme EOs showed the strongest activity against all tested isolates at low concentrations, followed by tea tree and peppermint EOs. The MIC value ranged from 0.06 µL/mL to 1.0 µL/mL. The tested EOs showed a significant antibiofilm activity against biofilm-forming isolates with MBIC50 ranging from 0.015 µL/mL to 0.25 µL/mL. All tested isolates were obtained from rainbow trout free of clinical signs of infection. Twelve isolates of Aeromonas salmonicida subsp. masoucida, four Aeromonas hydrophila, and four isolates of Aeromonas veronii were identified. The results of the in vitro study showed a significant effect of EOs against Aeromonas spp., which confirmed their potential for use in aquaculture as a prevention against bacterial diseases and a way of reducing the use of antibiotics.

Uptake, Translocation, Toxicity, and Impact of Nanoparticles on Plant Physiological Processes

The application of nanotechnology in agriculture has increased rapidly. However, the fate and effects of various nanoparticles on the soil, plants, and humans are not fully understood. Reports indicate that nanoparticles exhibit positive and negative impacts on biota due to their size, surface property, concentration within the system, and species or cell type under test. In plants, nanoparticles are translocated either by apoplast or symplast pathway or both. Also, it is not clear whether the nanoparticles entering the plant system remain as nanoparticles or are biotransformed into ionic forms or other organic compounds. Controversial results on the toxicity effects of nanomaterials on the plant system are available. In general, the nanomaterial toxicity was exerted by producing reactive oxygen species, leading to damage or denaturation of various biomolecules. The intensity of cyto- and geno-toxicity depends on the physical and chemical properties of nanoparticles. Based on the literature survey, it is observed that the effects of nanoparticles on the growth, photosynthesis, and primary and secondary metabolism of plants are both positive and negative; the response of these processes to the nanoparticle was associated with the type of nanoparticle, the concentration within the tissue, crop species, and stage of growth. Future studies should focus on addressing the key knowledge gaps in understanding the responses of plants to nanoparticles at all levels through global transcriptome, proteome, and metabolome assays and evaluating nanoparticles under field conditions at realistic exposure concentrations to determine the level of entry of nanoparticles into the food chain and assess the impact of nanoparticles on the ecosystem.

Genome-Wide Identification of the bHLH Gene Family in Callerya speciosa Reveals Its Potential Role in the Regulation of Isoflavonoid Biosynthesis

Callerya speciosa (Champ. ex Benth.) Schot is a significant leguminous plant valued for its edible tuberous roots, which are a plentiful source of isoflavonoids. Basic helix–loop–helix (bHLH) transcription factors (TFs) have been reported to regulate secondary metabolism in plants, especially flavonoid biosynthesis. However, the bHLH genes in C. speciosa have not yet been reported, and their regulatory role in isoflavonoid biosynthesis remains unexplored. Here, 146 CsbHLH genes were identified in the C. speciosa genome, classifying them into 23 subfamilies based on the gene structures and phylogenetic relationships. All the CsbHLH proteins contained both motifs 1 and 2, whereas motif 8 was only distributed in subgroup III (d + e). Collinearity analysis demonstrated that fragmental replications are the primary driver of CsbHLH evolution, with the majority of duplicated CsbHLH gene pairs experiencing selective pressure. Nine candidate CsbHLH genes were found to play a potential role in regulating isoflavonoid biosynthesis through a combination of gene-to-metabolite correlation analysis and weighted gene co-expression network analysis (WGCNA). Additionally, the cis-regulatory elements and response to MeJA of these nine genes were characterized and confirmed through quantitative real-time PCR (qRT-PCR) analysis. Among them, three CsbHLHs (CsbHLH9, CsbHLH89, and CsbHLH95) were selected for further investigation. Yeast two-hybrid (Y2H), dual-luciferase (LUC) assays, bimolecular fluorescence complementation (BiFC) assays, and transient transformation demonstrated that CsbHLH9 acted as a transcriptional activator through its interaction with CsMYB36 and binding to the promoters of isoflavonoid biosynthesis genes in a MeJA-induced manner, such as CsIFR2, CsI3′H2, and CsCHS4, to promote isoflavonoid (calycosin, calycosin-7-o-glucoside, and formononetin) accumulation. Our results establish a basis for the functional analysis of bHLH genes and investigations into the molecular mechanisms underlying isoflavonoid biosynthesis in C. speciosa.

A R2R3-MYB transcription factor, FeR2R3-MYB, positively regulates anthocyanin biosynthesis and drought tolerance in common buckwheat (Fagopyrum esculentum)

The R2R3-MYB transcription factors (TFs) play a crucial role in regulating plant secondary metabolism and abiotic stress responses, yet they are still poorly understood in common buckwheat (Fagopyrum esculentum), a valuable minor grain crop resource. In this study, a candidate gene, FeR2R3-MYB, was cloned from the anthocyanin-rich common buckwheat variety ‘QZZTQ’. FeR2R3-MYB was found to contain two MYB DNA-binding domains and be located at the nucleus with transcriptional activation activity. Molecular analysis indicated that FeR2R3-MYB is predominantly expressed in flowering tissue and is highly responsive to environmental factors such as light, drought, and cold. In addition, the promoter of FeR2R3-MYB showed a positive correlation with fragment length. Further functional analysis suggested that FeR2R3-MYB not only participates in the anthocyanin biosynthetic pathway by interacting with leucoanthocyanidin reductase (FeLAR), but also enhances drought tolerance in common buckwheat. To sum up, FeR2R3-MYB exhibits positive effects on both pigment production (e.g., anthocyanin) and abiotic stress resistance, providing valuable insights for future research in buckwheat molecular breeding and resource development.

Integrated Analysis of Metabolites and Microorganisms Reveals the Anthracnose Resistance Benefits from Cyanidin Mediated by Proteobacteria in Tea Plants.

Anthocyanins, key quality components of tea, act as an important bridge between plants and the environment due to their function on protecting plants from biotic and abiotic irritants. This study aimed to assess the interactions between anthocyanins metabolism and the environment. Purple (P) and green (G) leaves with different anthocyanin contents were inoculated with tea plant anthracnose. High-throughput metabolomics and 16S microbial diversity sequencing methods were used to screen the anthocyanin fractions of tea plant leaves responsive to anthracnose. The interconnections between metabolites and the resistance of phyllosphere microorganisms to fungal pathogens were then analyzed. The results showed that leaves with high anthocyanin content (0.14% of diseased area ratio) were less impacted by anthracnose infestation than leaves with low anthocyanin (3.12%). The cyanidin content decreased after infection in purple leaves (PR) and increased in green leaves (GR). The relative abundance of Cyanobacteria was suppressed by the significant enrichment of Proteobacteria after anthracnose infection in green leaves. However, there were no significant differences between these two groups of microorganisms in purple leaves. Collinear network analysis revealed a strong correlation between Cyanobacteria and Dihydrosorbinol and between Proteobacteria and cyanidin metabolites. Among them, OTU456 (Bosea) was identified as the key taxonomic group of bacterial communities in the green-infected leaf network. In summary, the anthracnose resistance benefits from cyanidin mediated by proteobacteria in tea plants. These results deepen our understanding of the regulation of secondary metabolism in tea plants and the formation of plant resistance.

Identification of Black Cumin (Nigella sativa) MicroRNAs by Next-Generation Sequencing and Their Implications in Secondary Metabolite Biosynthesis.

Secondary metabolites are bioactive compounds believed to contribute to the pharmacological properties of plants. MicroRNAs (miRNAs) are small non-coding RNA molecules involved in post-transcriptional regulation and are thought to play an important role in regulating secondary metabolism biosynthesis. Nevertheless, the extent of miRNA involvement in secondary metabolism remains minimal. Nigella sativa (black cumin/black seed) is a popular medicinal and culinary plant known for its pharmaceutical properties; however, its genomic information is scarce. In this study, next-generation sequencing (NGS) technology was employed to obtain the miRNA profile of N. sativa, and their involvement in secondary metabolite biosynthesis was explored. A total of 25,139,003 unique reads ranging from 16 to 40 nucleotides were attained, out of which 240 conserved and 34 novel miRNAs were identified. Moreover, 6083 potential target genes were recognized in this study. Several conserved and novel black cumin miRNAs were found to target enzymes involved in the terpenoid, diterpenoid, phenylpropanoid, carotenoid, flavonoid, steroid, and ubiquinone biosynthetic pathways, among others, for example, beta-carotene 3-hydroxylase, gibberellin 3 beta-dioxygenase, trimethyltridecatetraene synthase, carboxylic ester hydrolases, acetyl-CoA C-acetyltransferase, isoprene synthase, peroxidase, shikimate O-hydroxycinnamoyltransferase, etc. Furthermore, sequencing data were validated through qPCR by checking the relative expression of eleven randomly selected conserved and novel miRNAs (nsa-miR164d, nsa-miR166a, nsa-miR167b, nsa-miR171a, nsa-miR390b, nsa-miR396, nsa-miR159a, nsa-miRN1, nsa-miRN29, nsa-miRN32, and nsa-miRN34) and their expression patterns were found to be corroborated with the sequencing data. We anticipate that this work will assist in clarifying the implications of miRNAs in plant secondary metabolism and aid in the generation of artificial miRNA-based strategies to overproduce highly valuable secondary metabolites from N. sativa.

Effects of saline-alkali stress on cotton growth and physiochemical expression with cascading effects on aphid abundance.

Environmental stresses, such as soil salinity or alkalinity, usually affect crop growth and secondary plant metabolism, with follow on effects on foliar-feeding insects. Nevertheless, the underlying mechanism of how saline-alkali stress affects the key cotton pest Aphis gossypii Glover is poorly understood. In this study, we first considered effects of three types of saline-alkali stress (i.e., salinity alone, alkalinity alone - both at different concentration - and their mixed effects) on cotton plants. We then measured impacts of stress on (1) above and below plant growth traits (e.g., plant height, leaf area, root volume), (2) levels of nutrients and secondary metabolites in cotton leaves, and (3) feeding behavior, life-table parameters, and population growth of A. gossypii. We then used a path analysis to evaluate cascading effects of changes in plant growth (due to stress) and changes in levels of nutrients or secondary metabolites on growth of individual cotton aphids and aphid populations. We found either salinity or alkalinity stresses significantly reduced cotton growth, increased the content of tannin, soluble sugars, and proline in the leaves, and suppressed aphid growth and development, (including longevity, fecundity, and intrinsic rate of increase) and aphid population growth. Alkalinity had stronger effects on these traits than did salinity. This work provides insights into the bottom-up interaction mechanism by which these environmental stresses mediate aphid infestation levels in the cotton agricultural ecosystem.

Applications of Plant Essential Oils in Pest Control and Their Encapsulation for Controlled Release: A Review

Essential oils (EOs) are volatile products derived from the secondary metabolism of plants with antioxidant, antimicrobial, and pesticidal properties. They have traditionally been used in medicine, cosmetics, and food additives. In agriculture, EOs stand out as natural alternatives for pest control, as they show biocidal, repellent, and antifeedant effects. However, they are highly volatile compounds and susceptible to oxidation, which has limited their use as pesticides. This has led to exploring micro- and nano-scale encapsulation to protect these compounds, improving their stability and allowing for a controlled release. Various encapsulation techniques exist, such as emulsification, ionic gelation, and complex coacervation. Nanoemulsions are useful in the food industry, while ionic gelation and complex coacervation offer high encapsulation efficiency. Materials such as chitosan, gelatin-gum-Arabic, and cyclodextrins are promising for agricultural applications, providing stability and the controlled release of EOs. Encapsulation technology is still under development but offers sustainable alternatives to conventional agrochemicals. This article reviews the potential of EOs in pest management and encapsulation techniques that enhance their efficacy.

Evaluation of the Secondary Metabolites and Bioactivity of South African Bulbine Natalensis Under Simultaneous Elevated Carbon Dioxide and Temperatures

The secondary metabolism in medicinal plants are responsible for their protection against environmental factors and the health-promoting benefits they provide to users. The purpose of this study was to assess the responses of the secondary metabolic system of B. natalensis and its influence on the antioxidant and antibacterial activities. The phytochemical analyses, antioxidant and antibacterial assays were conducted from using harvested leaves, underground stems and roots from whole plants exposed to simultaneous elevated CO2 and temperatures over eight days in a heatwave simulation. Most of the phytochemical groups were present in the leaves, however majority of the tested compounds had consistent presence throughout the plants in all treatments. The underground stems possessed a greater total phenolic, tannin and proanthocyanidin contents than the leaves and roots under elevated CO2 and temperatures. The leaves had a comparatively better antioxidant activity from the treatment than control. Overall, there was consistency in the antibacterial activity in both control and experimental conditions. The tannins and phenolics had greater concentrations throughout the plants under elevated conditions overall, which could indicate their leading role in plant defence and the improvement and maintenance of the medicinal activity of B. natalensis. The responses of the plant parts of B. natalensis under elevated CO2 and temperatures provides newfound insights on the physiological roles plant organs play on the overall productivity and medicinal activity of the species.

Comparative genomics and transcriptomics analysis of the bHLH gene family indicate their roles in regulating flavonoid biosynthesis in Sophora flavescens.

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.

Comprehensive, Genome-Wide Identification and Expression Analyses of Phenylalanine Ammonia-Lyase Family under Abiotic Stresses in Brassica oleracea.

Phenylalanine ammonia-lyase (PAL) acts as the rate-limiting enzyme for anthocyanin biosynthesis through the phenylpropanoid pathway, a crucial component of plant secondary metabolism. The PAL gene family plays a crucial role in plants' defense and stress responses, but its in silico identification and expression analyses in Brassica oleracea under different abiotic stresses remain unexplored. In this study, nine BolPAL, seven BrPAL, four AtPAL, and seventeen BnPAL genes were obtained from the genomes of B. oleracea, Brassica rapa, Arabidopsis thaliana, and Brassica napus, respectively. Segmental duplication and purifying selection are the causes of the BolPAL gene's amplification and evolution. The BolPAL genes with comparable intron-exon architectures and motifs were grouped together in the same clade. Three categories comprised the cis-regulatory elements: abiotic stressors, phytohormones, and light. According to the results of the qRT-PCR experiments, the majority of the BolPAL genes were expressed highly under MeJA, a low temperature, and a high temperature, and they were downregulated under ABA. Under white light (100 µmol m-2 s-1) with 50, 100, or 150 µmol m-2 s-1 far-red (FR), only a small number of the PAL genes were expressed at 50 and 100 µmol m-2 s-1 FR, while the majority of the PAL genes were slightly elevated at 150 µmol m-2 s-1 FR. This work offers a theoretical foundation for molecular breeding research to investigate the role of BolPAL genes and their role in anthocyanin biosynthesis.

Mitigating gadolinium toxicity in guar (Cyamopsis tetragonoloba L.) through the symbiotic associations with arbuscular mycorrhizal fungi: physiological and biochemical insights

BackgroundGadolinium (Gd) is an increasingly found lanthanide element in soil; thus, understanding its impact on plant physiology, biochemistry, and molecular responses is crucial. Here, we aimed to provide a comprehensive understanding of Gd (150 mg kg− 1) impacts on guar (Cyamopsis tetragonoloba L.) plant yield and metabolism and whether the symbiotic relationship with arbuscular mycorrhizal fungi (AMF) can mitigate Gd toxicity of soil contamination.ResultsAMF treatment improved mineral nutrient uptake and seed yield by 38–41% under Gd stress compared to non-inoculated stressed plants. Metabolic analysis unveiled the defense mechanisms adopted by AMF-treated plants, revealing carbon and nitrogen metabolism adaptations to withstand Gd contamination. This included an increase in the synthesis of primary metabolites, such as total sugar (+ 39% compared to control), soluble sugars (+ 29%), starch (+ 30%), and some main amino acids like proline (+ 57%) and phenylalanine (+ 87%) in the seeds of AMF-treated plants grown under Gd contamination. Furthermore, fatty acid and organic acid profile changes were accompanied by the production of secondary metabolites, including tocopherols, polyamines, phenolic acids, flavones, and anthocyanins.ConclusionsOverall, the coordinated synthesis of these compounds underscores the intricate regulatory mechanisms underlying plant-AMF interactions and highlights the potential of AMF to modulate plant secondary metabolism for enhanced Gd stress tolerance.

Construction of the First High-Density Genetic Linkage Map and QTL Mapping of Shikimic Acid Content in Liquidambar

High-quality genetic maps are effective tools for elucidating the genetic mechanisms of complex quantitative traits and facilitating marker-assisted breeding. Species within the genus Liquidambar (commonly called sweetgum), particularly Liquidambar styraciflua and Liquidambar formosana, are significant forest resources worldwide. These sweetgum trees have been extensively utilized in medical and cosmetic applications for centuries as they contain large amounts of valuable secondary metabolites. Among these, shikimic acid is a notable metabolite with significant pharmaceutical applications. Despite advances in conventional breeding and propagation techniques for sweetgum, the genetic basis and regulatory mechanisms of valuable traits remain largely unexplored. In this study, we constructed the first high-density genetic map for sweetgum using whole-genome resequencing (WGR) of 220 progeny individuals derived from a cross of L. styraciflua × L. formosana. The genetic map spanned a total distance of 1428.51 centimorgans (cM) with an average inter-marker distance of 0.33 cM, incorporating 4268 bin markers across 16 linkage groups. To identify the genetic loci controlling the shikimic acid content, quantitative trait locus (QTL) mapping was carried out based on the genetic map. Two QTLs located on linkage group (LG) 12 were detected, encompassing a total of 213 genes within the QTL interval. Some of these genes are closely related to secondary metabolism in plants, including YUCCA and DXS genes. This study presents the first high-quality genetic map of sweetgum and provides a preliminary QTL analysis for shikimic acid content. Our findings establish a foundational framework for the genetic improvement of sweetgum through marker-assisted breeding and offer valuable insights for further research in sweetgum genetics.

Enzymatic Characterization of SpPAL Genes in S. polyrhiza and Overexpression of the SpPAL3.

Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) catalyzes the deamination of phenylalanine, which is the initial step in the biosynthesis of phenylpropanoids. It serves as a crucial enzyme that facilitates the transfer of carbon from primary to secondary metabolism in plants. Duckweed is regarded as a promising chassis plant in synthetic biology research and application, due to its being rich in secondary metabolites and other advantages. The genes encoding PAL in Spirodela polyrhiza (L.) Schleid, the giant duckweed, were investigated in this study. Three SpPAL genes (SpPAL1-SpPAL3) were identified and cloned. All of them were successfully expressed in E. coli, and their recombinant proteins all showed PAL activity. In addition, SpPAL1 and SpPAL2 proteins could also utilize tyrosine as substrate, although the activity was low. A qRT-PCR analysis demonstrated that the expression of SpPAL3 was most pronounced in young fronds. It was found that the expression of SpPAL1 and SpPAL3 was significantly induced by MeJA treatment. Overexpression of SpPAL3 in Lemna turionifera inhibited the growth of fronds and adventitious roots in the transgenic plants, indicating the importance of SpPAL3 in duckweed besides its involvement in the secondary metabolism.

Integrated HS-SPME-GC–MS and UHPLC-O-TOF-MS-based metabolomics revealed the characteristics flavor and nonvolatile metabolites during the hydrolysis process of Boletus edulis

This research aimed to investigate the flavor characteristic, metabolites and bioactivities of Boletus edulis hydrolysates via HS-SPME-GC-MS and UHPLC-MS/MS. The Maillard reaction (MR) significantly enhanced the sensory qualities of Boletus edulis, including like-meat, caramel, continuity, and complete acceptance. The accumulation of browning precursors during MR was verified through spectral analysis. We also found that the MR enhanced the antioxidant and hypoglycemic activities of Boletus edulis hydrolysates. Furthermore, 11 VOCs were also identified as key aromatic compounds and had important impact on the flavor characteristics of Maillard reaction products (MRPs). Non-targeted metabolomics analyses revealed that the formation of MRPs was affected through amino acid metabolism, lipid metabolism and plant secondary metabolism. Briefly, the flavor and biological activity of Boletus edulis were improved through enzymolysis and MR. The results provided a theoretical foundation for deep processing and resource utilization of Boletus edulis.

BZIP transcription factor responds to changes in light quality and affects saponins synthesis in Eleutherococcus senticosus

Light quality considerably influences plant secondary metabolism, yet the precise mechanism underlying its impact on Eleutherococcus senticosus remains elusive. Comprehensive metabolomic and transcriptomic analyses revealed that varying light quality alters the biosynthesis of triterpene saponins by modulating the expression of genes involved in the process in E. senticosus. Through correlation analysis of gene expression and saponin biosynthesis, we identified four light-responsive transcription factors, namely EsbZIP1, EsbZIP2, EsbZIP4, and EsbZIP5. EsbZIP transcription factors function in the nucleus, with light quality-dependent promoter activity. Except for EsbZIP2, the other EsbZIP transcription factors exhibit transcriptional self-activation. Furthermore, EsbZIP can bind to the promoter areas of genes that encode important enzymes (EsFPS, EsSS, and EsSE) involved in triterpene saponin biosynthesis, thereby regulating their expression. Overexpression of EsbZIP resultes in significant down-regulation of most downstream target genes，which leads to a decrease in saponin content. Overall, varying light quality enhances the content of triterpene saponins by suppressing the expression of EsbZIP. This study thus elucidates the molecular mechanism by which E. senticosus adjusts triterpene saponin levels in response to changes in light quality.

Phytochemical Screening and Biological Activity (Antioxidant and Antibacterial) of Punica granatum Fruit peel extract Cultivated in Sa'dah Governorate-Yemen

Punica granatum (Pomegranate) is a valuable source of secondary plant metabolism compounds (SMs) that are widely used in the pharmaceutical, cosmetics, and food industries. The phytochemical screening illustrates that the Sa’dah P. granatum cultivar peel ethanol extract containsvarious beneficial components, including Flavonoids, Alkaloids, Steroids, Phenols, and Tannins. Moreover, the Sa’dah P. granatum cultivar peel ethanol extract (1.3 μg/ml) exhibited significant antioxidant activity compared to low concentrations of ascorbic acid (0.4, 0.6, 0.8, and 1 μg/ml). Furthermore, the Sa’dah P. granatum cultivar peel ethanol extract displays potent antibacterial activities against selected pathogenic bacteria (Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa) compared to two nominated antibiotics (Ampicillin and Gentamicin). This remarkable performance can be attributed to the presence of phenols and flavonoid compounds, which exhibit high antioxidant activity.