Cinnamate 4-hydroxylase Research Articles

Combined physiological, biochemical, and multi-omic analyses provide insight into the cause of differential browning in early and late harvested fresh-cut yams

Tissue browning reduces the cosmetic quality of fresh-cut yams and poses a marketing problem. The degree of browning of fresh-cut yams varies depending on when they are harvested. In this study, transcriptomic and metabolomic analyses were conducted in yams harvested in September (DHPS) and October (DHPO) to determine the mechanisms responsible for the browning of fresh-cut yams. Results showed that the browning index, total phenol and lignin content, polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia-lyase (PAL), 4-coumarate: CoA Ligase(4CL), and cinnamic acid-4-hydroxylase(C4H) activity in fresh-cut DHPO yams were lower than they were in fresh-cut DHPS yams. Flavonoids and catalase (CAT) activity remained consistently higher during storage in DHPO yams than in DHPS yams. The browning of fresh-cut DHPO yams was mitigated by the regulation of the expression of genes and the abundance of metabolites associated with phenolic metabolism, phytohormones, and signal transduction, as well as sucrose and starch metabolism. The level of browning enzyme activity in DHPO yams was significantly lower than it was in DHPS yams. Our study provides a basis for the selection of optimum harvest date for yams that will undergo fresh-cut processing.

Effects of sucrose and 1-MCP on enzymatic and nonenzymatic antioxidants in postharvest Gynura bicolor DC

After harvesting, Gynura bicolor DC (G. bicolor) undergoes rapid quality deterioration, including decay, nutrient loss, and reactive oxygen species (ROS) burst, greatly limiting its shelf life. This study was performed to evaluate the effects of treatment with sucrose and 1-methylcyclopropene (1-MCP) on indices of quality deterioration, ROS metabolism, and phenylpropanoid metabolism, with the goals of resisting oxidative stress and improving the postharvest quality of G. bicolor. Sucrose treatment activated phenylpropanoid metabolism, increased phenylalanine ammonia lyase (PAL), cinnamate-4-hydroxylase (C4H), and 4-coumarate:CoA ligase (4CL) activities, and promoted the accumulation of phenolics. The increases in nonenzymatic antioxidants decreased O2·−, ·OH, and H2O2 contents. Conversely, 1-MCP treatment increased NADPH oxidase (NOX) activity and induced transient oxidative stress, which significantly activated enzymatic antioxidants, including superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX), thus maintaining a lower ROS level. Treatment with both sucrose and 1-MCP exhibited a synergistic effect on alleviating oxidative stress, downregulating GbSAG101 and GbATG expression. Compared with sucrose treatment, 1-MCP treatment showed a greater inhibitory effect on ROS burst. Phenolic substance contents were similar in plants treated with sucrose and 1-MCP at the end of the storage period. Therefore, we speculated that higher levels of antioxidant enzyme activity may decrease the consumption of nonenzymatic antioxidants. Our experimental results indicated the involvement of different pathways in the effects of sucrose treatment, 1-MCP treatment, and combined treatment on delaying ROS burst, and suggested potential alternative methods for the preservation of vegetables with stems during the postharvest storage.

Ultraviolet-B Radiation Stimulates Flavonoid Biosynthesis and Antioxidant Systems in Buckwheat Sprouts.

Abiotic stress not only elevates the synthesis of secondary metabolites in plant sprouts but also boosts their antioxidant capacity. In this study, the mechanisms of flavonoid biosynthesis and antioxidant systems in buckwheat sprouts exposed to ultraviolet-B (UV-B) radiation were investigated. The findings revealed that UV-B treatment significantly increased flavonoid content in buckwheat sprouts, with 3-day-old sprouts exhibiting a flavonoid content 1.73 times greater than that of the control treatment. UV-B radiation significantly increased the activities of key enzymes involved in flavonoid biosynthesis (phenylalanine ammonia-lyase, 4-coumarate-CoA ligase, cinnamate 4-hydroxylase, and chalcone synthase) and the relative expression levels of the corresponding genes. Although UV-B radiation caused damage to the cell membranes of buckwheat sprouts, promoting increases in hydrogen peroxide and malondialdehyde content and inhibiting the growth of sprouts, importantly, UV-B radiation also significantly increased the activities of catalase, peroxidase, and superoxide dismutase as well as the relative expression levels of the corresponding genes, thus enhancing the antioxidant system of buckwheat sprouts. This enhancement was corroborated by a notable increase in ABTS, DPPH, and FRAP radical scavenging activities in 3-day-old sprouts subjected to UV-B radiation. Additionally, UV-B radiation significantly increased chlorophyll a and chlorophyll b contents in sprouts. These results suggest that UV-B radiation is advantageous for cultivating buckwheat sprouts with increased flavonoid content and enhanced antioxidant capacity.

Manipulating flavonoid biosynthesis in Trigonella persica through controlled spectral lighting

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

Investigation of the association between gene expression levels and phenolic compound content in the leaves of Sonchus arvensis plants under salinity stress

Sonchus arvensis is recognized for its high content of phenolic compounds and antioxidants, exhibiting various medicinal benefits, including anti-diabetic, anti-depressant, and anti-cancer properties. This has positioned the plant as a significant candidate for commercial food, medicinal, and antioxidant supplements. Salinity may enhance the level of chlorogenic and caffeic acid, which are key secondary metabolites in S. arvensis. To investigate this, a completely randomized design experiment with three replications was implemented in a greenhouse to examine the impact of salinity on the expression of six genes responsible for the biosynthesis of chlorogenic acid. Additionally, the study examined how salinity affects the accumulation of chlorogenic acid, caffeic acid, chicoric acid, and apigenin in the lower and middle leaves of plants. Salinity stress treatments were applied at four different levels: 0 (control), 50, 100, and 150 mM of NaCl. The results indicated that the expression levels of phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), and p-coumaroyl ester 3′-hydroxylase (C3′H) were highest in the middle leaves at a concentration of 150 mM NaCl. Notably, there was an eight-fold increase in C4H expression in these leaves under the same salinity conditions. Conversely, the expression of shikimate/quinate O-hydroxycinnamoyl transferase (HCT) and quinate O-hydroxycinnamoyl transferase (HQT) genes decreased across all salinity treatments. Additionally, the levels of chlorogenic acid, caffeic acid, and chicoric acid were significantly elevated at 50 mM NaCl in both the lower and middle leaves, suggesting that cultivating S. arvensis in mildly saline environments could be beneficial. Furthermore, the findings from this study may serve as a preliminary step towards the cloning and full characterization of the genes examined in S. arvensis.

Multiple NADPH-cytochrome P450 reductases from Lycoris radiata involved in Amaryllidaceae alkaloids biosynthesis.

Amaryllidaceae alkaloids (AAs), such as galanthamine and lycorine, are natural products of Lycoris radiata possessing various pharmacological activities including anti-acetylcholinesterase, anti-inflammatory, and antitumour activities. Elucidating the biosynthesis of these special AAs is crucial for understanding their production and potential modification for improved clinical application, of which cytochrome P450 enzymes catalyse the formation of key alkaloid skeletons and subsequent modification processes, with the NAPDH cytochrome P450 reductases (CPRs) serving as essential redox partners. This study identified three CPRs, LrCPR1, LrCPR2, and LrCPR3, encoding 700, 697 and 695 amino acids, respectively, which belong to Class II CPRs. The LrCPRs reduced cytochrome c and ferricyanide in an NADPH-dependent manner, and their activities all followed the typical Michaelis-Menten curve. In yeast, the co-expression of LrCPRs and CYP96T6 produced the galantamine-like alkaloid namely N-demethylnarwedine, suggesting that they support the catalytic activity of CYP96T6. Quantitative analysis of the transcriptional expression profiles showed that LrCPRs were expressed in all the examined tissues of L. radiata, and their gene expression patterns are consistent with other genes that may be involved in the biosynthetic pathway of AAs, including cinnamate 4-hydroxylase and phenylalanine ammonia-lyase. Our study firstly provides the functional characterization of LrCPRs in L. radiata, which will contribute to the discovery of biosynthetic pathways and heterologous production of AAs.

Dielectric Barrier Discharge Cold Plasma Improves Storage Stability in Paddy Rice by Activating the Phenylpropanoid Biosynthesis Pathway.

A nonthermal pretreatment using dielectric barrier discharge cold plasma (DBD-CP) was developed to improve the stress resistance of paddy rice during postharvest storage. The physicochemical properties, bioactive characteristics, and secondary metabolites of paddy rice were assessed after applying an optimized DBD-CP procedure, with enzyme activities and gene expression monitored over a 60 day storage period at 35 °C. A 17.06% reduction in the total color change index was noted in the DBD-CP group. Bioactive compounds, particularly gallic acid, were significantly increased, enhancing the defense mechanisms against high-temperature stress. Nontargeted metabolomics analysis indicated an upregulation of phenylpropanoid metabolism in DBD-CP-treated rice compared to controls, with notable increases in secondary metabolites such as coumaric acid, caffeic acid, and sinapic acid, suggesting potential biomarkers for stress resistance. Further verification showed significant enhancements in key enzymes of phenylpropanoid metabolism, including phenylalanine ammonia lyase (PAL), cinnamic acid-4-hydroxylase (C4H), plant coumaric acid-3-hydroxylase (C3H), and cinnamyl alcohol dehydrogenase (CAD), with increases ranging from 1.71 to 2.28 times. Gene expression levels of OsPAL7, OsC4H4, and OsCAD2 aligned with these enzymatic changes post-DBD-CP treatment. In conclusion, DBD-CP treatment can modulate phenylpropanoid metabolism in paddy rice, thereby enhancing bioactive compound levels to reduce stress damage during high-temperature storage.

Integrated Analysis of Metabolome and Transcriptome Reveals the Effect of Burdock Fructooligosaccharide on the Quality of Chinese Cabbage (Brassica rapa L. ssp. Pekinensis).

Burdock fructooligosaccharide (BFO) is fructose with a low polymerization degree, which could improve the immunity to pathogens, quality, and stress resistance of vegetables. Still, there are no studies on applying BFO in Chinese cabbage. In this study, the effects of exogenous BFO sprayed with different concentrations (0, 5, 10, 20, 30 g·L-1) on the growth and soluble sugar content of Chinese cabbage seedlings were determined. The result showed that 10 g·L-1 was the appropriate spraying concentration. Based on metabolome analysis, a total of 220 differentially accumulated metabolites (DAMs) were found, among which flavonoid metabolites, glucosinolate metabolites, and soluble sugar-related metabolites were the key metabolites involved in improving the quality of Chinese cabbage caused by BFO. Further combination analysis with transcriptome, trans-cinnamate 4-monooxygenase (CYP73A5), and chalcone synthase 1 (CHS1) were more closely associated with the DAMs of flavonoid biosynthesis. Sulfotransferases 18 (SOT18), Branched-chain amino acid amino transferases 6 (BCAT6), and cytochrome P450 monooxygenase (CYP83A1) were the key genes in glucosinolate biosynthesis. Hexokinase (HxK1), beta-glucosidase 8 (BGL08), invertase 3 (INV3), beta-glucosidase 3B (BGL3B), and sucrose phosphate synthase 1 (SPS1) were significantly upregulated, potentially playing crucial roles in the soluble sugar metabolism. In conclusion, these results provided an understanding of the effects of BFO on the expression of genes and the accumulation of metabolites related to quality formation in Chinese cabbage.

Functional analysis of HpMYB72 showed its positive involvement in high-temperature resistance via repressing the expression of HpCYP73A

Global warming has led to occasional occurrence of extreme climates, especially summer high temperature, which constrains crops in growth and development. Pitaya (Hylocereus polyrhizus) characterizes in excellent tolerance to high temperature and drought stresses. Previously, a differentially expressed R2R3-MYB transcription factor gene, designated as HpMYB72, was identified in pitaya as exposure to high temperature. To elucidate the function of HpMYB72, currently, its spatiotemporal expression, as well as the roles involving in high-temperature response and regulating the downstream genes were conducted. The data from RT-qPCR and GUS reporter system showed that HpMYB72 was mainly expressed in rapidly developing tissues, particularly, in the flowers. It was activated by high temperature, and the coded proteins were accumulated in nucleus, indicating that HpMYB72 may act as a positive regulator involved in the high-temperature response of pitaya. Ectopic overexpression of HpMYB72 in Arabidopsis thaliana improved the growth performance and increased the germination rate as exposure to 42 °C; reduced the accumulation of reactive oxygen species under high-temperature stress thus alleviated the oxidative damage; increased the content of osmotic regulators, thereby relieved the water loss as exposure to high temperature. The available evidences comprehensively reflected that overexpression of HpMYB72 improved the tolerance of Arabidopsis against high temperature, supporting the hypothesis that HpMYB72 is a positive regulator of high-temperature response. The results of yeast one-hybrid assay and luciferase reporter system demonstrated that HpMYB72 bound directly to the promoter of pitaya cinnamate 4-hydroxylase gene HpCYP73A and inhibited its transcription. In this way, the NADPH which ought to be the substrate of cinnamate 4-hydroxylase was reassigned to ROS scavenging, and led the conversion of cinnamoyl-CoA to pinocembrin chalcone, which protects pitaya flowers from malformation under high temperature.

Comparative transcriptomic and metabolomic analysis reveals mechanisms of selenium-regulated anthocyanin synthesis in waxy maize (Zea mays L.).

Anthocyanins in maize (Zea mays L.) kernels determine the plant's color and can enhance its resistance. Selenium (Se) significantly impacts plant growth, development, and secondary metabolic regulation. However, the molecular mechanisms by which Se regulates anthocyanin synthesis in waxy corn remain unclear. This study employed integrated transcriptomic and metabolomic analyses to investigate the mechanisms through which selenium influences anthocyanin synthesis in yellow and purple waxy corn. The results showed that maize varieties with higher anthocyanin content had higher selenium enrichment capacity in their kernels. Under selenium stress, HN2025 exhibited 1,904 more differentially expressed genes (DEGs) and 140 more differential metabolites compared to HN5. The expression levels of anthocyanin synthesis-related genes and transcription factors such as phenylalanine ammonia-lyase, flavonoid 3-hydroxylase (F3H), dihydroflavonol reductase (DFR), chalcone synthase (CHS), cinnamate-4-hydroxylase (C4H), anthocyanin 5,3-O-glucosyltransferases, and anthocyanidin reductase, MYB, and bHLH were strongly induced in HN2025. Metabolomic analysis revealed significant enrichment in anthocyanin biosynthesis, flavonoid and flavonol biosynthesis, glutathione metabolism, phenylalanine biosynthesis, and phenylalanine metabolism under selenium treatment. Three up-regulated PAL genes and one C4H gene were significantly enriched with DAMs in phenylalanine metabolism, phenylpropanoid biosynthesis, flavonoid biosynthesis, and anthocyanin biosynthesis, resulting in significant differences between HN5 and HN2025 in selenium-induced anthocyanin metabolism-related pathways. These findings provide a theoretical basis for understanding the effects of selenium on the molecular regulatory mechanisms of anthocyanin biosynthesis in maize kernels.

Exogenous p-coumaric acid treatment improves phenolic compounds biosynthesis of postharvest cherry tomatoes by regulating physiological metabolism.

To investigate the effect of p-coumaric acid (p-CA) on postharvest cherry tomatoes, breaker stage fruits were treated with p-CA to analyze the physiological metabolism during storage. The results showed that exogenous p-CA treatment improved the sensory quality of tomato fruits. Transcriptomics results indicated that 782 genes (339 up-regulated and 443 down-regulated) were differentially expressed between p-CA treated and control fruits. Results suggested that p-CA treatment regulated the synthesis of phenolic compounds and inhibited the fruit ripening through the pathways of mangiferic acid biosynthesis and phenylalanine metabolism. Key enzymes activities of the phenylpropane metabolic pathway of tomato fruits were increased, including phenylalanine ammonia lyase (PAL), cinnamic acid 4-hydroxylase (C4H), and 4-coumarate-CoA ligase (4CL). In addition, the total phenols content, flavonoids content and antioxidant capacities of tomato fruits were improved with p-CA treatment. Overall, these findings showed that p-CA treatment could be a potential strategy for fruit and vegetable preservation.

Metabolome and Transcriptome Joint Analysis Reveals That Different Sucrose Levels Regulate the Production of Flavonoids and Stilbenes in Grape Callus Culture.

To reveal the effect of sucrose concentration on the production of secondary metabolites, a metabolome and transcriptome joint analysis was carried out using callus induced from grape variety Mio Red cambial meristematic cells. We identified 559 metabolites-mainly flavonoids, phenolic acids, and stilbenoids-as differential content metabolites (fold change ≥2 or ≤0.5) in at least one pairwise comparison of treatments with 7.5, 15, or 30 g/L sucrose in the growing media for 15 or 30 days (d). Resveratrol, viniferin, and amurensin contents were highest at 15 d of subculture; piceid, ampelopsin, and pterostilbene had higher contents at 30 d. A transcriptome analysis identified 1310 and 498 (at 15 d) and 1696 and 2211 (at 30 d) differentially expressed genes (DEGs; log2(fold change) ≥ 1, p < 0.05) in 7.5 vs. 15 g/L and 15 vs. 30 g/L sucrose treatments, respectively. In phenylpropane and isoflavone pathways, DEGs encoding cinnamic acid 4-hydroxylase, chalcone synthase, chalcone isomerase, and flavanone 3-hydroxylase were more highly expressed at 15 d than at 30 d, while other DEGs showed different regulation patterns corresponding to sucrose concentrations and cultivation times. For all three sucrose concentrations, the stilbene synthase (STS) gene exhibited significantly higher expression at 15 vs. 30 d, while two resveratrol O-methyltransferase (ROMT) genes related to pterostilbene synthesis showed significantly higher expression at 30 vs. 15 d. In addition, a total of 481 DEGs were annotated as transcription factors in pairwise comparisons; an integrative analysis suggested MYB59, WRKY20, and MADS8 as potential regulators responding to sucrose levels in flavonoid and stilbene biosynthesis in grape callus. Our results provide valuable information for high-efficiency production of flavonoids and stilbenes using grape callus.

Decoding eggplant fruit: Multi-omics profiling of caffeoyl-CoA-3-OMT expression

Eggplant (Solanum melongena L.) is a rich source of health-promoting phenolic acids, mainly chlorogenic acid (CGA), and trace elements. CGA has been shown to have antioxidant, anti-inflammatory, neuroprotective, cardioprotective, anticancer, and antidiabetic properties. The natural genome of eggplants contains essential genes for beneficial traits like drought tolerance, disease resistance, and high concentrations of medicinal substances. Transcriptomic analysis of eggplant overexpressing the hydroxycinnamoyl CoA quinate hydroxycinnamoyl transferase (HQT) gene revealed upregulation of 2165 genes, including three similar to HQT and cinnamate-4-hydroxylase (C4H). Sequence similarity analysis showed homology to caffeoyl-CoA O-methyltransferases, key enzymes in the phenylpropanoid pathway leading to CGA biosynthesis. Gene ontology and KEGG pathway analysis identified 5 highly upregulated glycosyltransferase family 43 genes (650.3-fold change). Glycosyltransferases like UDP-glucose:cinnamate glucosyltransferase and cinnamate-glucose 4′-O-glucosyltransferase are crucial for CGA production in eggplant. qRT-PCR validated the potential upregulation of HQT and C4H in transgenic eggplants. Comparative analysis revealed eggplant has the highest CGA content (5–8.1 g/kg dry weight) among vegetables, with foliar CGA acting as a natural insecticide. CGA offers antioxidant, hypoglycemic, antiviral, and hepatoprotective benefits to humans. This study highlights the role of HQT and C4H in elevating the medicinal properties of eggplant through CGA biosynthesis pathway engineering.

Inhibitory effect of oxyresveratrol on the browning and PPO of fresh-cut potato

The work investigated the browning inhibition of oxyresveratrol (OXY) in fresh-cut potatoes by physicochemical and structural analysis. The results showed that the optimal inhibitory concentration was 0.002 % for fresh-cut potato sticks. Under this condition, oxyresveratrol inhibited the activities of polyphenol oxidase (PPO), peroxidase (POD), cinnamate-4-hydroxylase (C4H) and catalase (CAT), and decreased the content of chlorogenic acid and caffeic acid, but increased the contents of catechic acid and epicatechin in fresh-cut potatoes. Further analysis found that oxyresveratrol was a competitive inhibitor for PPO with the strong ability to chelate Cu2+. Oxyresveratrol formed hydrogen bonds with Ala246, Ala143, Met247 and Ala146 of StPPO2, and constructed stable hydrophobic interactions with His144, Pro245, Glu145 and Ile151 residues. The analysis of tertiary and secondary structures showed that oxyresveratrol decreased the fluorescence intensity and α-helix content of PPO, but increased β-sheet and β-turn content. The results showed that oxyresveratrol is a promising anti-browning agent for fresh-cut potatoes.

Global organization of phenylpropanoid and anthocyanin pathways revealed by proximity labeling of trans-cinnamic acid 4-hydroxylase in Petunia inflata petal protoplasts.

The phenylpropanoid pathway is one of the main carbon sinks in plants, channeling phenylalanine towards thousands of products including monolignols, stilbenes, flavonoids and volatile compounds. The enzymatic steps involved in many of these pathways are well characterized, however the physical organization of these enzymes within the plant cell remains poorly understood. Proximity-dependent labeling allows untargeted determination of both direct and indirect protein interactions in vivo, and therefore stands as an attractive alternative to targeted binary assays for determining global protein-protein interaction networks. We used TurboID-based proximity labeling to study protein interaction networks of the core phenylpropanoid and anthocyanin pathways in petunia. To do so, we coupled the endoplasmic reticulum (ER) membrane anchored cytochrome P450 cinnamic acid 4-hydroxylase (C4H, CYP73A412) from Petunia inflata to TurboID and expressed it in protoplasts derived from anthocyanin-rich petunia petals. We identified multiple soluble enzymes from the late anthocyanin pathway among enriched proteins, along with other C4H isoforms, and other ER membrane anchored CYPs. Several of these interactions were subsequently confirmed by bimolecular fluorescence complementation (BiFC). Our results suggest that C4H co-localizes with enzymes from the phenylpropanoid- and downstream anthocyanin pathways, supporting the idea that C4H may serve as ER anchoring points for downstream metabolic pathways. Moreover, this study demonstrates the feasibility of using protoplasts to perform global mapping of protein network for enzymes in their native cellular environment.

Metabolomic and transcriptomic analyses reveal the mechanism of polysaccharide and secondary metabolite biosynthesis in Bletilla striata tubers in response to shading

Polysaccharides and various secondary metabolites are the major bioactive ingredients in Bletilla striata tubers and their biosynthesis and accumulation are influenced by light intensity. However, the mechanisms underlying shading effects remain largely unknown. In the present study, we used a combined analysis of the physiology, metabolome, and transcriptome to investigate the physiological activities and bioactive component accumulation of B. striata under different shading treatments (S0, S50, S70, and S90). The dry weight of shoots and tubers, net photosynthetic rate, and polysaccharide content were highest in S50 and lowest in S90. The content of precursors (sucrose, Glucose-6P, and Mannose-6P) for polysaccharide synthesis significantly increased in S50. However, the expression levels of genes involved in starch biosynthesis decreased in S50. Several structural genes involved in secondary metabolism, including cinnamic acid 4-hydroxylase (C4H), chalcone synthase (CHS), and 1-Deoxy-D-xylulose-5-phosphate synthase (DXS), showed decreased expression in S50. However, the shading effect on the biosynthesis of secondary metabolites (phenylpropanoids, flavonoids, and terpenoids) was inconsistent. Our study provides the molecular mechanisms underlying the effects of shading on the biosynthesis of polysaccharides and secondary metabolites in B. striata and offers a theoretical basis for the artificial cultivation and industrial production of bioactive ingredients.

Metagenomic and transcriptome insights into the disease resistance mechanisms in fresh-cut apple under pyridoxine treatment

Microbial spoilage poses a significant challenge to fresh-cut apples during storage, impacting quality and consumer health. This study explores pyridoxine’s (vitamin B6; VB6) potential mode of action to mitigate microbial spoilage in fresh-cut apples through physiological, transcriptomic, and metagenomic analysis. VB6 notably reduces microbial spoilage at room temperature by suppressing bacterial genera (Pseudomonas, Curtobacterium, Frigoribacterium) and fungal genera (Alternaria, Penicillium, Aureobasidium, Pseudogymnoascus) proliferation. Spoilage-causing identified biomarkers include species related to Pseudomonas, Curtobacterium, Frigoribacterium, and Penicillium. KEGG analysis suggests correlations of induced disease resistance with tropane alkaloids biosynthesis, plant-pathogen interaction, and phenylpropanoid biosynthesis pathways in fresh-cut apples. Moreover, VB6 treatment enhanced the antioxidant system via maintaining higher total phenolic contents, total flavonoid contents, and ascorbic acid under elevated catalase, superoxide dismutase, and ascorbate peroxidase enzymes activities, induced disease resistance under higher enzymes activities including phenylalanine ammonialyase, chitinase, cinnamate 4-hydroxylase, β-1,3-glucanase, and 4-coumarate: co-enzyme A ligase enzymes activities, maintained elevated VB6 contents, and reduced H2O2 accumulation. Notably, 6 elevated transcription factors involving WRKY, NAC and MYB might regulate disease resistance pathways in fresh-cut apples under VB6 treatment. This research establishes a theoretical and practical foundation for preserving post-processing quality of fresh-cut apples by restraining microbial proliferation through the proposed VB6-induced disease resistance regulatory mechanism.

Hydrogen Sulfide Improves Postharvest Quality of Okra (Abelmoschus esculentus (L.) Moench) Pods by Enhancing Antioxidant Capacity and Delaying Lignification.

Okra (Abelmoschus esculentus (L.) Moench) pod storage is challenging due to its high water content and tendency to lignify. Sodium hydrosulfide (NaHS) served as an H2S donor in this investigation. Compared with the control group, the group treated with 0.5 mmol/L NaHS solution effectively maintained the appearance quality, and its weight loss was only 6.21% at 20 days. The H2S treatment not only preserved tissue nutrients but also significantly enhanced catalase (CAT), ascorbic acid peroxidase (APX), and superoxide dismutase (SOD) activities while decreasing oxidant damage. In addition, H2S slowed down lignin synthesis by inhibiting the activities of key enzymes such as phenylalanine ammonialyase (PAL), cinnamate 4-hydroxylase (C4H), and cinnamyl alcohol dehydrogenase (CAD) in the lignin biosynthesis pathway. Transcriptome analysis revealed that H2S affects 34 genes in the phenylpropanoid biosynthesis pathway, such as AePAL, Ae4CL1, AeCCOAOMT1, AePOD, etc., which inhibit lignin synthesis of okra pods. All in all, moderate H2S can improve postharvest quality and extend the shelf-life of okra pods by enhancing antioxidant capacity and delaying lignification; the results will provide an overview of its application in the preservation of okra pods.

The Impact of Lighting Treatments on the Biosynthesis of Phenolic Acids in Black Wheat Seedlings.

Light, as a crucial environmental determinant, profoundly influences the synthesis of secondary metabolites in plant metabolism. This study investigated the impacts of the red light combined with ultraviolet-A (UV-A) and ultraviolet-B (UV-B) treatments on phenolic acid biosynthesis in black wheat seedlings. The results demonstrate that the red light combined with UV-A and UV-B treatments significantly enhanced the levels of phenolic acids in black wheat seedlings, at 220.4 μg/seedling and 241.5 μg/seedling, respectively. The content of bound phenolic acids in black wheat seedlings increased by 36.0% under the UV-B treatment. The application of the UV-A/UV-B treatments markedly enhanced the activities of phenylalanine ammonia-lyase, 4-coumarate CoA ligase, and cinnamate 4-hydroxylase in black wheat seedlings while also promoting the expression levels of genes related to phenolic acid synthesis. The expression levels of fructose-1,6-bisphosphate aldolase and NADP-malic enzyme related to photosynthesis were significantly upregulated. This resulted in an augmentation in the chlorophyll content, thereby enhancing photosynthesis in black wheat seedlings. Nevertheless, the UV-A and UV-B treatments also had a significant constraining effect on the growth and development of black wheat seedlings. In addition, the UV-A and UV-B treatments increased the activity and gene expression levels of antioxidant enzymes while significantly increasing the contents of total flavonoids and anthocyanins, activating the antioxidant system. The findings reveal that light-source radiation serves as an effective method for promoting the biosynthesis of phenolic acids in black wheat seedlings.

Germination Promotes Flavonoid Accumulation of Finger Millet (Eleusine coracana L.): Response Surface Optimization and Investigation of Accumulation Mechanism.

Germination is an effective measure to regulate the accumulation of secondary metabolites in plants. In this study, we optimized the germination conditions of finger millet by response surface methodology. Meanwhile, physiological characteristics and gene expression were measured to investigate the mechanism of flavonoid accumulation in finger millet at the germination stage. The results showed that when germination time was 5.7 d, germination temperature was 31.2 °C, and light duration was 17.5 h, the flavonoid content of millet sprouts was the highest (7.0 μg/sprout). The activities and relative gene expression of key enzymes for flavonoid synthesis (phenylalanine ammonia-lyase, 4-coumarate-coenzyme a ligase, and cinnamate 4-hydroxylase) were significantly higher in finger millet sprouts germinated at 3 and 5 d compared with that in ungerminated seeds (p < 0.05). In addition, germination enhanced the activities of four antioxidant enzymes (catalase, peroxidase, superoxide dismutase, and ascorbate peroxidase) and up-regulated the gene expression of PAL and APX. Germination increased malondialdehyde content in sprouts, which resulted in cell damage. Subsequently, the antioxidant capacity of the sprouts was enhanced through the activation of antioxidant enzymes and the up-regulation of their gene expression, as well as the synthesis of active substances, including flavonoids, total phenolics, and anthocyanins. This process served to alleviate germination-induced cellular injury. These findings provide a research basis for the regulation of finger millet germination and the enhancement of its nutritional and functional properties.