Micro(nano)plastics (MNPs) often coexist with antibiotics in soil ecosystems via organic fertilizer applications. They can be taken up and accumulated by plants, especially food crops, thereby threatening food safety. De...Micro(nano)plastics (MNPs) often coexist with antibiotics in soil ecosystems via organic fertilizer applications. They can be taken up and accumulated by plants, especially food crops, thereby threatening food safety. Despite increasing global concern, the distinct phytotoxic mechanisms of MNPs and their interactive dynamics with antibiotics remain dispersed. First, the uptake, translocation, and distribution of MNPs in plants are systematically synthesized. Based on the clarification of plant phenotypic and physiological responses to MNP stress, the review assesses their biochemical and molecular mechanisms, ranging from oxidative stress and antioxidant defense to signal transduction and transcriptional regulation to metabolic trade-offs. After clarifying the physicochemical interactions and cotransport mechanisms between MNPs and antibiotics, the review elucidates their combined phytotoxicity and their crucial role in antibiotic resistance gene (ARG) transmission in soil-plant systems. Finally, we identify research gaps and propose future research directions, which will facilitate the evaluation of ecological risks and the formulation of environmental regulations for copollutants.
Tomato () sepal morphology impacts fruit external quality, yet its genetic regulatory mechanism remains unclear. Accordingly, we constructed a segregating population from noncurling (P47) and inward-curling (AC) inbred...Tomato () sepal morphology impacts fruit external quality, yet its genetic regulatory mechanism remains unclear. Accordingly, we constructed a segregating population from noncurling (P47) and inward-curling (AC) inbred lines, and fine-mapped the major sepal curvature locus to a 25-kb distal region of chromosome 4 via BSA-seq and genetic mapping. Four candidate genes were found in this interval, with only harboring a parental-specific Ser9Ala nonsynonymous SNP cosegregating with the trait in cultivated germplasm. This Type I MADS-box transcription factor gene was designated (sepal curvature regulator). expression was significantly lower in AC than in P47. knockout induced inward curling in noncurling sepals of 5P and aggravated inherent sepal curvature in AC, alongside reduced auxin, gibberellin, and cytokinin. These findings establish as a critical regulator of tomato sepal curvature, delivering theoretical underpinnings for sepal development and genetic resources for trait-targeted tomato breeding.
Naringenin, a chiral flavonoid, contains a chiral center and exists as ()- and ()-enantiomers, yet its enantiomer-specific metabolism has remained largely unexplored. Herein, high-purity ()- and ()-naringenin were isolat...Naringenin, a chiral flavonoid, contains a chiral center and exists as ()- and ()-enantiomers, yet its enantiomer-specific metabolism has remained largely unexplored. Herein, high-purity ()- and ()-naringenin were isolated via chiral semipreparative chromatography, and their racemization kinetics under near-physiological conditions were systematically quantified. Incubation with rat liver microsomes revealed pronounced stereoselectivity: the ()-enantiomer was metabolized significantly faster than the ()-enantiomer. Mechanistic studies identified the CYP3A subfamily as the key mediator of this stereoselectivity. Notably, this stereoselective profile was completely absent in human liver microsomes, uncovering a critical species difference. These findings provide direct evidence for enzyme-level chiral recognition in naringenin metabolism and highlight species-specific disparities in its metabolic pathway.
3,4-Dimethylpyrazole phosphate (DMPP) is an effective nitrification inhibitor, but its poor thermal stability limits its performance in fertilizer formulations and soil systems. In this study, DMPP was conjugated with γ-...3,4-Dimethylpyrazole phosphate (DMPP) is an effective nitrification inhibitor, but its poor thermal stability limits its performance in fertilizer formulations and soil systems. In this study, DMPP was conjugated with γ-polyglutamic acid (γ-PGA) via a proposed covalent linkage to enhance its stability and nitrification inhibition efficacy in soil. The optimized DMPP-γ-PGA conjugate formed a compact three-dimensional network with a DMPP loading of 85.1%. Thermal analyses showed that γ-PGA conjugation reduced DMPP loss by 72.6% after heating at 200 °C for 30 min. Soil incubation experiments showed that the DMPP-γ-PGA conjugate (1:10) sustained nitrification inhibition (average 46.8% over 21 days) and reduced cumulative NO emissions by 50.1% relative to urea alone, outperforming free DMPP at the same application rate (45.4%). The improved performance was attributed to enhanced thermal stability and controlled release of DMPP in soil. This study provides a biodegradable polymer-inhibitor conjugation strategy for improving nitrification inhibitor functionality in agricultural systems.
The oleaginous yeast is an outstanding chassis for producing high-value chemicals. Its highly active β-oxidation pathway generates a substantial pool of acetyl-CoA within peroxisomes; however, this abundant reservoir re...The oleaginous yeast is an outstanding chassis for producing high-value chemicals. Its highly active β-oxidation pathway generates a substantial pool of acetyl-CoA within peroxisomes; however, this abundant reservoir remains largely underutilized. Here, we developed a peroxisomal surface-display platform in , enabling enhanced biosynthesis of acetyl-CoA-derived chemicals. We first identified three peroxisomal membrane-targeting signals, derived from Pex3, Pex22, and Pex26 for protein display on the cytosolic face of the peroxisome. By anchoring the carnitine acetyl-CoA transferase Cat2 via these membrane-targeting signals, peroxisomal acetyl-CoA was redirected to the cytosol, increasing acetyl-CoA level and mevalonate production up to 2.4-fold and 2.9-fold, respectively. Further engineering improved squalene titers by 47% to 1.62 g/L. The same platform applied to 3-hydroxypropionic acid biosynthesis improved its production by 22% to 12.85 g/L. Our study establishes a peroxisomal surface display platform for harnessing subcellular acetyl-CoA and provides a versatile toolkit for subcellular engineering in .
Anthocyanin biosynthesis is tightly controlled by the MYB-bHLH-WD40 (MBW) complex in plants. Here, we investigated the regulatory mechanism underlying anthocyanin accumulation in passion fruit ( Sims) peels. We identifie...Anthocyanin biosynthesis is tightly controlled by the MYB-bHLH-WD40 (MBW) complex in plants. Here, we investigated the regulatory mechanism underlying anthocyanin accumulation in passion fruit ( Sims) peels. We identified PebHLH42, a nucleus-localized IIIf bHLH transcription factor that interacts with activator PeMYB114. The PeMYB114-PebHLH42 complex strongly activates and transcription, promoting anthocyanin accumulation in tobacco leaves, whereas PebHLH42 alone shows no inductive effect. We also characterized PeMYB6, a nuclear R2R3-MYB repressor that harbors EAR and TLLLFR motifs. PeMYB6 overexpression reduces the anthocyanin and proanthocyanidin levels and represses flavonoid biosynthetic genes. Mechanistically, PeMYB6 competes with PeMYB114 for binding to PebHLH42, thus suppressing the activation of and . Moreover, PeMYB114 activates PeMYB6 transcription, forming an activator-repressor feedback loop. These results reveal a refined regulatory module for passion fruit pigmentation and benefits molecular breeding for color modification.
Food-grade strategies against foodborne pathogens while preserving intestinal health are gaining attention. Here, we developed a regulated delivery platform using engineered to secrete a tandem dimeric antimicrobial pep...Food-grade strategies against foodborne pathogens while preserving intestinal health are gaining attention. Here, we developed a regulated delivery platform using engineered to secrete a tandem dimeric antimicrobial peptide (SD). The engineered strain (SDLactis) exhibited stable growth, genetic stability, and tolerance to simulated gastrointestinal conditions. Chloride-responsive SD secretion enabled effective antibacterial activity against enterotoxigenic (ETEC) K88 . In ETEC-challenged piglets, oral SDLactis alleviated diarrhea, improved growth, and reduced intestinal injury and inflammation. It strengthened intestinal barrier integrity by upregulating tight junction proteins (ZO-1, Claudin-1,Occludin) and downregulating CFTR. Microbiome analysis revealed that SDLactis partially restored gut microbial diversity, reducing while enriching and short-chain fatty acid-producing genera. Overall, food-grade engineered lactic acid bacteria serve as controllable delivery vehicles for antimicrobial peptides, offering a nonantibiotic strategy for pathogen control and gut health management in food and feed applications.
Cadmium (Cd) contamination in threatens food safety and aroma quality. We compared titanium dioxide nanoparticle (TiO) and methyl jasmonate (MeJA) effects singly or combined using sensory evaluation, HS-SPME-GC-MS, indu...Cadmium (Cd) contamination in threatens food safety and aroma quality. We compared titanium dioxide nanoparticle (TiO) and methyl jasmonate (MeJA) effects singly or combined using sensory evaluation, HS-SPME-GC-MS, inductively coupled plasma mass spectrometry (ICP-MS), and transcriptome analysis. Cd stress reduced pleasant floral and fruity notes, increased irritating odors, elevated floral Cd accumulation, and shifted volatiles to defense-related sesquiterpenes (e.g., α-cadinol, γ-/d-cadinene). MeJA enriched α-farnesene, enhancing fresh green attributes. ICP-MS revealed TiO and MeJA each reduced floral Cd by approximately 50%, while their combination achieved 88.3% reduction. Transcriptomics showed Cd upregulated phenylpropanoid/flavonoid genes (, , , , ) and activated ABC transporters and salicylic acid (SA) axis (). TiO attenuated ABCB and ABCG expression, and MeJA enhanced α-linolenic acid metabolism and jasmonate signaling (via ). Weighted gene coexpression network analysis (WGCNA) and gene set enrichment analysis (GSEA) indicated energy, redox, and terpenoid pathways (e.g., , ) were involved. This study provides a theoretical basis for synergistic safety and flavor of aromatic crops in heavy-metal-contaminated areas.
The widely used deworming drugs nitroxynil (NIT) and oxyclozanide (OXY) pose potential risks to the ecological environment and human health, necessitating a sensitive and rapid detection method. In this study, two novel...The widely used deworming drugs nitroxynil (NIT) and oxyclozanide (OXY) pose potential risks to the ecological environment and human health, necessitating a sensitive and rapid detection method. In this study, two novel haptens targeting OXY and NIT, respectively, were designed and synthesized, with their structural validity evaluated by computer simulation. Through animal immunization and cell fusion, monoclonal antibodies (OXY-1A4 and NIT-2F12) were obtained, with half-maximal inhibitory concentrations of 1.13 ng/mL and 0.23 ng/mL, respectively. Based on this, a multiplex gold nanoparticle-based immunochromatographic assay (multi-AuNPs-ICA) was established for simultaneous detection of OXY and NIT in wastewater, milk, and beef. Visual and calculated limits of detection ranged from 2 to 20 ng/mL and 0.37-7.16 ng/mL, respectively. Spiked recovery rates in wastewater, milk, and beef samples were 102.6%-110.5%, 90.4%-115.6%, and 86.9%-117.7%, respectively. These were highly consistent with instrumental methods, confirming the reliability of multi-AuNPs-ICA and its significant potential for on-site rapid screening.
Global health challenges, including the growing burden of chronic diseases and persistent micronutrient deficiencies, underscore the need for novel functional resources. Rosehip, the fruit of the spp. has a rich history...Global health challenges, including the growing burden of chronic diseases and persistent micronutrient deficiencies, underscore the need for novel functional resources. Rosehip, the fruit of the spp. has a rich history of medicinal and nutritional use, suggesting its potential as an underused source of diverse bioactive compounds. However, research on rosehips remains scattered, with limited systematic integration of their ethnomedicinal uses, bioactive compounds, and therapeutic potentials. This review critically synthesizes recent advances in the identification of rosehip bioactive compounds, their pharmacological activities, traditional medicinal applications, and available clinical safety evidence. The review also identifies the primary constraints affecting the broader application of rosehips, including inefficient resource utilization, fragmented evidence, limited clinical validation, and weak commercialization pathways. The objective is to critically evaluate the existing evidence to assess the potential for developing functional foods and therapeutic products based on rosehips.
Epigallocatechin gallate (EGCG), the most abundant and bioactive catechin in green tea, has limited applications due to its chemical instability, low solubility, and poor bioavailability. Enzymatic glycosylation offers a...Epigallocatechin gallate (EGCG), the most abundant and bioactive catechin in green tea, has limited applications due to its chemical instability, low solubility, and poor bioavailability. Enzymatic glycosylation offers a sustainable method to improve these properties. However, the regioselectivity and catalytic efficiency of enzymatic glycosylation remain challenging. In the present study, a loop-exchanged variant of sucrose phosphorylase from (SP), designated SP_LoopB, shifted the major product from EGCG-4'--α-d-glucopyranoside to EGCG-4″--α-d-glucopyranoside. The SP_LoopB I235S/V297A mutant, generated through a tailored, tunnel-focused mutagenesis strategy, exhibited a 140.5-fold increase in catalytic efficiency compared to SP_LoopB. Under optimized reaction conditions (15 g/L EGCG, 336 g/L sucrose, 2 U/mL enzyme), the yield of EGCG-4″--α-d-glucopyranoside reached 83.3% (16.9 g/L) within 4 h. The mutant also demonstrated excellent performance and stability in the whole-cell biocatalytic system. This work establishes a synergistic engineering approach that enhances specificity and efficiency, enabling the scalable production of well-defined EGCG glycosides.
A key challenge in enzyme engineering is coordinating local catalytic dynamics with global structural integrity. Here, we present a hierarchical strategy integrating local dynamics optimization with global scaffold rigid...A key challenge in enzyme engineering is coordinating local catalytic dynamics with global structural integrity. Here, we present a hierarchical strategy integrating local dynamics optimization with global scaffold rigidification, using d-erythrose-4-phosphate dehydrogenase (Epd) as a model. Guided by evolutionary and conformational network analyses, we first engineered local active-site dynamics, yielding a double mutant (G14A/A234S) with 3-fold higher and improved thermostability (Δ = 2 °C). Global rigidification further empowered the preoptimized active site, producing a quadruple mutant (G14A/A234S/T31I/V17I) with 11.2-fold increased and an additional +2 °C rise. Molecular dynamics simulations revealed that distal rigidification suppresses nonproductive fluctuations and enriches catalytically competent conformations. This "inner flexibility, outer rigidity" architecture boosted vitamin production by 3.9-fold. Our work establishes a mechanism-guided paradigm for synergistically enhancing both activity and stability, offering a generalizable framework for engineering industrial biocatalysts.
Honey bees rely on olfaction to perceive pheromones and environmental chemicals. Odorant-binding proteins (OBPs) are key carriers in odorant recognition, while growing studies suggest that OBPs may function beyond chemos...Honey bees rely on olfaction to perceive pheromones and environmental chemicals. Odorant-binding proteins (OBPs) are key carriers in odorant recognition, while growing studies suggest that OBPs may function beyond chemosensation, particularly in response to chemical stress. In this study, expression in the antennae of was significantly upregulated after sublethal exposure to flonicamid (FLO) or its metabolite 4-(trifluoromethyl) nicotinamide (TFNA-AM). Fluorescence competitive binding assays showed that AmOBP18 exhibited high binding affinity for TFNA-AM ( = 0.65 ± 0.01 μM) and FLO ( = 1.14 ± 0.03 μM). Surface plasmon resonance (SPR) analysis confirmed these findings, with a value of 0.163 μM for TFNA-AM. Moreover, overexpression of in significantly enhanced bacterial tolerance to TFNA-AM. Silencing of increased honey bee sensitivity to both FLO and TFNA-AM. We demonstrate that AmOBP18 may have functions beyond its classical olfactory role, contributing to tolerance-related responses to flonicamid and TFNA-AM.
Serotonin (5-hydroxytryptamine, 5-HT) plays a critical role in maintaining intestinal health, regulating mood and sleep, and enhancing crop stress tolerance. Here, a cell factory was developed for the sustainable and ef...Serotonin (5-hydroxytryptamine, 5-HT) plays a critical role in maintaining intestinal health, regulating mood and sleep, and enhancing crop stress tolerance. Here, a cell factory was developed for the sustainable and efficient synthesis of 5-HT. First, a tryptophan-producing chassis strain, TRP18, was obtained by blocking competitive pathways, modifying the tryptophan operon, and rewiring carbon metabolic flux, and the Trp titer was increased by 23.94-fold over the starting strain, TRP0. Subsequently, a heterologous 5-HT biosynthetic pathway was established, and tryptophan hydroxylase Luz15 was optimized by protein engineering. Finally, the 5-HT pathway was optimized by a dual-cassette system, and an optimal strain, HT-12, was attained, producing 1.23 g/L 5-HT in a 5-L bioreactor, representing the highest titer of 5-HT from glucose in species to date. This study provides an efficient and sustainable process for 5-HT production in , laying the foundation for industrial-scale manufacturing of 5-HT and other Trp-derived biochemicals.
Low-temperature stress seriously affects the growth and development of maize seedlings, endangering their quality and yield. In this study, the low-temperature tolerance at the seedling stage of 299 maize inbred lines wa...Low-temperature stress seriously affects the growth and development of maize seedlings, endangering their quality and yield. In this study, the low-temperature tolerance at the seedling stage of 299 maize inbred lines was evaluated by using the membership function method. A genome-wide association study identified 12 SNPs associated with low-temperature tolerance. By integrating GWAS, RNA-seq, and metabolomics data, we identified a gene 3 encoding an E3 ubiquitin-protein ligase. Its involvement in low-temperature tolerance during seedling development was further supported by the RT-qPCR analysis. 3 may regulate the accumulation of secondary metabolites, influencing the plant hormone and membrane-associated signaling. Functional evidence from the ethylmethanesulfonate (EMS) mutant 3 showed markedly reduced tolerance to low-temperature stress. 3 acts in synergy with 147 and may be involved in the regulation of low-temperature tolerance in maize seedlings. These support its role in promoting seedling adaptation and deepen the current understanding of cold response pathways in maize.
The oriental fruit moth (), a worldwide fruit tree pest, can be effectively controlled by (Bt), with the insect peritrophic membrane (PM) serving as a key target for such control. This study investigates the role of the...The oriental fruit moth (), a worldwide fruit tree pest, can be effectively controlled by (Bt), with the insect peritrophic membrane (PM) serving as a key target for such control. This study investigates the role of the key PM protein ATP synthase subunit b (GmolATPs-b) in the molecular mechanism of Cry2Ab toxin toxicity. was found to be highly expressed in fourth larval midguts, and its expression is downregulated upon Cry2Ab exposure. Ligand blotting, competitive experiments, and molecular docking confirmed specific binding between GmolATPs-b and activated Cry2Ab. Heterologous expression in Sf9 cells enhanced susceptibility to Cry2Ab, whereas larval RNAi knockdown reduced it. assays showed that cofeeding purified GmolATPs-b with Cry2Ab increased larval mortality. This study demonstrates that GmolATPs-b is a functional target of Cry2Ab, binding specifically to the toxin and enhancing its insecticidal effect, thus establishing a molecular basis for novel biopesticides against .
Citrus canker, caused by subsp. (), is a devastating bacterial disease. Given the resistance and pollution from long-term chemical pesticides, plant-derived natural products offer green alternatives. Here, daurichromen...Citrus canker, caused by subsp. (), is a devastating bacterial disease. Given the resistance and pollution from long-term chemical pesticides, plant-derived natural products offer green alternatives. Here, daurichromenic acid (DAA) from var. showed potent anti- activity with an MIC of 10 μg/mL, which is 1/16 that of copper sulfate. Physiological assays revealed that DAA rapidly damages cell membranes and causes content leakage. Further mechanistic investigations, integrating metabolomics, qRT-PCR, and molecular modeling, revealed that DAA exerts a multitarget synergistic mode of action. DAA compromises membrane integrity as the primary lethal effect while interfering with the ABC transporter pathway, leading to ATP depletion, energy metabolism disorders, and suppressed virulence (biofilm formation and motility). Serial passage confirmed a low resistance risk, outperforming copper-based agents. Detached leaf and earthworm toxicity tests validated its disease control efficacy and environmental safety. Thus, DAA is a promising green pesticide for sustainable citrus canker management.
Vibration stress during transportation represents a mechanical factor that accelerates texture deterioration. This study revealed the molecular basis underlying vibration-induced texture deterioration in kiwifruit and fu...Vibration stress during transportation represents a mechanical factor that accelerates texture deterioration. This study revealed the molecular basis underlying vibration-induced texture deterioration in kiwifruit and further evaluated 1-methylcyclopropene (1-MCP) as an intervention. The results showed vibration accelerated pectin solubilization and nanostructural disassembly, whereas 1-MCP treatment better preserved pectin integrity with higher chelate-soluble pectin and sodium-carbonate-soluble pectin levels under vibration stress. Transcript analysis indicated that vibration induced elevated expression of pectin-degrading and -modifying genes, including polygalacturonase (PG) (Actinidia28962), β-galactosidase (Actinidia15368), and pectin methylesterase (PME) (Actinidia13568), whereas 1-MCP treatment downregulated these genes. Molecular docking indicated favorable binding poses of demethylesterified GalA oligomers with PME and higher-degree of polymerization GalA oligomers with PG, providing auxiliary insight into vibration-related pectin de-esterification and depolymerization. Therefore, this study provides a multilevel molecular explanation for vibration-induced texture deterioration from the perspective of pectin composition and structure and proposes a feasible strategy to delay texture loss under vibration stress.
Copper (Cu) is widely used as a growth-promoting trace element in swine feed, but excessive Cu causes liver toxicity and modulates innate immunity, with unclear molecular mechanisms. In this study, dietary Cu overload in...Copper (Cu) is widely used as a growth-promoting trace element in swine feed, but excessive Cu causes liver toxicity and modulates innate immunity, with unclear molecular mechanisms. In this study, dietary Cu overload in pigs led to Cu accumulation and liver injury, elevated levels of reactive oxygen species (ROS), and decreased mitochondrial membrane potential (MMP). These changes were accompanied by increased VDAC oligomerization in the mitochondrial membrane and mitochondrial DNA (mtDNA) release, which activates the NLRP3 inflammasome and triggers hepatocyte pyroptosis. Mechanistically, the VDAC inhibitor VBIT-4 reduced Cu-induced VDAC oligomerization and mtDNA release. Furthermore, NLRP3 is essential for Cu-mediated pyroptosis. Combined VBIT-4 and the MCC950 (NLRP3 inhibitor) treatment further attenuated VDAC oligomerization, mtDNA release, and pyroptosis. Collectively, our results reveal that Cu exposure promotes VDAC oligomerization and mitochondrial membrane pore formation, leading to mtDNA release, NLRP3 inflammasome activation, and pyroptosis in hepatocytes. These findings provide new insights into Cu-induced hepatotoxicity.
Cow's milk-based infant formula aims to approximate human milk, but bovine α-casein remains an important allergen-related protein with a profile distinct from human milk caseins. This study controlled trypsin hydrolysis...Cow's milk-based infant formula aims to approximate human milk, but bovine α-casein remains an important allergen-related protein with a profile distinct from human milk caseins. This study controlled trypsin hydrolysis to preferentially reduce intact α-casein and generate candidate bioactive peptides. Under 38 °C, pH 8.0, and 5 h, the relative residual peak area of α-casein decreased markedly, while α-casein and β-casein remained detectable at 33.15% and 45.01%, respectively. Peptidomics identified 17 peptides, among which HQGLPQEVLNENLLR showed the strongest predicted binding affinity to α-amylase by molecular docking (-13.98 kcal/mol). The hydrolysate showed ABTS radical scavenging activity of 92.10% and α-amylase inhibition of 53.44%. Molecular dynamics further supported the stability of the peptide-enzyme complex. These findings provide a preliminary process window for reducing intact α-casein and prioritizing candidate functional peptides from casein hydrolysates.