Plant volatile organic compounds (VOCs) represent one of the most dynamic and integrative biochemical signaling systems linking molecular plant stress responses to ecosystem-level processes. This review provides an integrative cross-scale framework for understanding the biochemical pathways, regulatory networks, ecological functions, and technological applications of stress-induced volatile emissions. At the molecular and cellular levels, VOC emissions are regulated through complex enzymatic and hormonal pathways involving jasmonates, salicylates, ethylene, and abscisic acid, enabling plants to respond rapidly to abiotic and biotic stressors such as drought, herbivory, temperature extremes, salinity, and atmospheric pollution. These volatile signals extend beyond individual plants, functioning as mediators of plant-plant communication, plant-microbe interactions, and multi-trophic ecological networks that shape community dynamics and ecosystem resilience. Recent technological advancements, including mass spectrometry platforms, remote sensing systems, biosensors, and artificial intelligence-driven analytical frameworks, have transformed the ability to detect, interpret, and predict stress-induced VOC emissions in real time. Integrating these technologies with multi-omics datasets and digital twin modeling enables the development of predictive monitoring systems capable of scaling plant stress detection from agricultural fields to regional ecosystems. Despite these advances, significant challenges remain, including variability in emission profiles across species and environments, atmospheric transformation of volatile signals, methodological inconsistencies, and limitations in large-scale monitoring infrastructure. Future research should focus on establishing global networks for monitoring plant volatiles, standardized measurement protocols, and integrated biosensing infrastructures that can link plant stress signals to Earth-system observations. Decoding plant volatile stress signaling across scales offers a transformative pathway to advance climate-resilient agriculture, biodiversity conservation, and predictive environmental intelligence systems that support adaptive ecosystem management in an era of accelerating environmental change.
Aspilia africana (Pers.) C.D.Adams is an important medicinal plant in Africa. In a previous study, we induced direct somatic embryogenesis (SE) from its leaf explants, by combining plant hormones with adenosine 5'-monophosphate (AMP) and nicotinamide adenine dinucleotide (NAD). SE is a valuable biotechnological tool with various applications such as synthetic seed production, and understanding molecular events that induce SE is vital for its effective manipulation and improvement. Therefore, this study aimed to elucidate the molecular basis of SE induced by a combination of plant hormones and other mitogenic molecules (AMP and NAD) in A. africana. Using NovaSeq 6000 (Illumina, Inc., USA) technology, we conducted denovo transcriptome sequencing of leaf tissues representing three distinct SE developmental stages: non-embryonic leaf (NEL), globular embryo formed leaf (GEL), and cotyledonary embryo formed leaf (CEL). A total of 8669 genes were differentially expressed in the three samples, and transcripts encoding plant hormone signalling components including auxin and cytokinin were markedly enriched. SE candidate genes such as SERK2, ERF12, ERF2, and LEA14-A were up-regulated in embryogenic tissues. Additionally, qRT-PCR analysis confirmed the expression levels of some selected SE related genes. For the first time, this study reports a de novo transcriptome assembly and gene expression analysis during SE induction in A. africana. The work also provides insights into SE induced by a combination of plant hormones with other mitogenic molecules, and findings suggest that the mechanism of SE induction in this case may be the same or similar to that induced by only plant hormones. Overall, this study offers beneficial resources for future molecular research on A. africana, and may be a key resource platform for future functional research in plant embryogenesis.
The plant cell wall not only serves as a physical barrier against pathogens but, when damaged, also functions as a source of cell wall-derived molecules that play crucial roles in plant immunity as damage-associated molecular patterns (DAMPs). While oligogalacturonides from homogalacturonan are well-studied DAMPs, the immune-signaling potential of other cell wall components remains largely unexplored. Conventional genetic and biochemical approaches aimed at identifying ligand-receptor pairs in plant immunity have been limited by the vast diversity of potential ligand molecules and functional redundancy of putative receptors. Here, we developed a high-throughput screening pipeline that simultaneously examines multiple interactions between plant cell wall-derived glycans and >350 extracellular domains (ECDs) of receptor kinases and receptor-like proteins in Arabidopsis, resulting in the screening of >40,000 interactions. We discovered a group of leucine-rich repeat receptor kinases named ARMs (AWARENESS of RG-I MAINTENANCES) that interact with rhamnogalacturonan-I (RG-I), a major component of pectin. RG-I treatment induced pattern-triggered immunity responses, with distinct kinetics compared to oligogalacturonide responses. We identified RG-I oligosaccharide structures required for interaction with ARM receptors and immune activation, and found that ARM receptors are redundantly involved in plant immunity. Collectively, our approach provides a powerful platform for discovering glycan-receptor pairs in plants, facilitating a more comprehensive understanding of cell wall surveillance mechanisms in plant immunity.
Some plant-beneficial microbes, including bacteria and fungi, can induce plant defense, enabling plants to resist pathogen infections. Successful defense elicitation depends on compatible host-microbe interactions at multiple stages. The initial interaction begins with plant root exudates, which contain chemical cues that attract beneficial microbes by enhancing their motility, biofilm formation, and expression of symbiosis- and immunity-related genes. In turn, these microbes produce a diverse array of immune elicitors-such as proteins, carbohydrates, lipids, and volatile compounds-that are perceived by plants through various mechanisms. Some elicitors are recognized by membrane-bound pattern recognition receptors, whereas others interact with the plant plasma membrane or cytoplasmic targets such as MYB72 and LOX3. These interactions can either trigger local pattern-triggered immunity, characterized by reactive oxygen species production and activation of the mitogen-activated protein kinase signaling pathway, or generate long-distance signals such as oxylipins that induce systemic resistance in distal tissues. A central outcome of these interactions is induced systemic resistance, which primes plants for a heightened immune state, enabling faster and stronger defense responses upon a subsequent pathogen challenge. In some cases, beneficial microbes can also trigger salicylic acid-mediated systemic acquired resistance, particularly enhancing resistance against biotrophic pathogens. Furthermore, beneficial microbes must balance immune activation and immune evasion by suppressing microbe-associated molecular pattern-triggered immune responses and avoiding the formation of hyperbiofilm, which allows them to establish a long-term symbiotic relationship with the host.
The GRF-Interacting Factor (GIF) family comprises plant-specific transcriptional coactivators essential for regulating diverse aspects of plant growth, development, and stress responses. Although the GIF gene family has been successively identified and studied in many plants, there have been no reports on its research in Zanthoxylum armatum. In this study, we identified five ZaGIF genes in Zanthoxylum armatum. Phylogenetic analysis revealed that ZaGIFs exhibit the closest evolutionary affinity with ZbGIFs from congeneric Zanthoxylum bungeanum, followed by CsGIFs from Citrus sinensis, indicating lineage-specific conservation within Rutaceae. ZaGIFs showed preferential expression in actively proliferating tissues, including buds, young flowers and young leaves, and were transcriptionally induced by auxin, abscisic acid and drought stress. We further performed comprehensive functional characterization of ZaGIF5. Subcellular localization indicated that ZaGIF5 predominantly accumulates in the nucleus, consistent with its role as a transcriptional coactivator. Heterologous overexpression of ZaGIF5 significantly promoted vegetative growth and markedly enhanced drought tolerance in Nicotiana benthamiana and Arabidopsis thaliana. Mechanistically, ZaGIF5 operates through a dual-layer regulatory mechanism. It transcriptionally upregulates AtGRF1 and AtGRF3, and physically interacts with these transcription factors to enhance their transcriptional activation activity, thereby orchestrating the modulation of downstream growth- and stress-responsive gene networks. Collectively, our study identifies ZaGIF5 as an evolutionarily conserved regulator that coordinates growth promotion and drought tolerance, offering a promising molecular target for the synergistic improvement of stress resilience and vegetative productivity in plants. However, its native functional validation in Zanthoxylum armatum or other woody plants such as poplar remains to be further confirmed in future studies.
Viral infections continue to pose a significant health problem to all parts of the world because of a high rate of mutation and the ability to resist traditional single-target antivirals. Multi-target inhibition of core viral proteins (main protease, RNA-dependent RNA polymerase, and spike glycoprotein) with plant metabolites as the target of drug discovery is a promising alternative. This review presented a systematic review of the antiviral potential and mechanism of plantderived compounds published from 2020 to 2025. The systematic search of the literature was performed with the help of PubMed, Scopus, Web of Science, Science Direct, and EMBASE in accordance with PRISMA 2020. Peer-reviewed articles that reported precise molecular targets, quantifiable antiviral activity (IC50, EC50 or %inhibition), and mechanistic information were included. Qualitative synthesis of data was done because of heterogeneity in methodology. Sixty studies were eligible. Flavonoids (quercetin, kaempferol) had an inhibitory effect on viral protease and polymerase with an IC50 value of 2-12 μM. The terpenoids (glycyrrhizin, betulinic acid) inhibited spike-ACE2 binding and viral fusion with up to 80% infectivity in vitro. The polymerases of hepatitis and influenza viruses were inhibited by polyphenols (EGCG, resveratrol). The bioavailability increased 150-200% with nano formulations, which enhanced therapeutic potency. Metabolites of plants have direct antiviral and host immunomodulatory properties. Nevertheless, a lack of clinical validation and inconsistency in the standardization of phytochemicals are critical issues. Plant-based metabolites are highly promising, multi-target antiviral leads. Nanotechnology, together with standardized pharmacological assessment and enhanced regulatory validation, has the potential to hasten clinical translation of plant metabolites into safe, broad-spectrum antiviral therapies.
Understanding the genetic diversity of the highly valued multipurpose tree species, drumstick, is crucial for effective breeding and conservation efforts. To evaluate the genetic diversity among 23 drumstick genotypes using both morphological and molecular markers. In a field study, eleven qualitative and twenty-one quantitative morphological traits were evaluated. Additionally, sixty-five genomic microsatellite (SSR) markers were utilized for molecular characterization, among which forty-four demonstrated polymorphism. The morphological study uncovered both qualitative and quantitative differences among the genotypes. Correlation and path analysis identified key traits that influence yield, i.e weight of individual pods and the no. of pods per plant. Genetic divergence analysis based on morphological data grouped the genotypes into six distinct clusters, highlighting significant inter-cluster distances that could be leveraged for heterosis breeding. Molecular marker analysis using the 44 polymorphic SSRs demonstrated substantial genetic diversity, with an average of 5.09 alleles per locus and a mean PIC of 0.57. Observed heterozygosity was relatively low (0.09) compared to expected heterozygosity (0.59), suggesting potential inbreeding or population structure. Phylogenetic analysis based on SSR data using a neighbor-joining tree clustered the 23 genotypes into three main groups, revealing distinct genetic divergence. The combined insights from morphological and molecular analyses provide a comprehensive understanding of the genetic diversity within the studied drumstick germplasm, offering valuable information for targeted breeding strategies and the effective conservation of this important species.
Phytohormonal integration, rather than isolated hormone action, coordinates salinity and temperature-stress resilience through shared signaling hubs, cross-tolerance, and translational crop-improvement strategies. Salinity and temperature extremes, as abiotic stresses, affect over 20% of global arable land, reducing crop yields by up to 50% under severe conditions and greatly threatening food security. Phytohormones, which include abscisic acid, auxins, cytokinins, gibberellins, salicylic acid, jasmonates, and ethylene, are small signaling molecules that play central roles in mediating plant adaptation and resilience under such stresses. This review critically examines the mechanisms by which phytohormones regulate plant responses to salinity and temperature stress. Specifically, physiological adjustments, molecular signaling pathways, and cross-talk interactions are discussed. The phytohormone-mediated modulation of osmotic balance and ion homeostasis, reactive oxygen species scavenging, stress-responsive gene expression, and hormonal priming underpins plant tolerance, often improving survival rates by 20-40% under experimental stress. Furthermore, integrative signaling mechanisms that contribute to cross-tolerance are discussed, along with practical applications, such as exogenous hormone treatment, breeding strategies, and genetic engineering, aimed at developing climate-resilient crops. To guide future studies, emerging research directions, such as multi-omics approaches, CRISPR-based manipulation, and computational modeling, are highlighted. This review provides a comprehensive framework for leveraging phytohormonal regulation to enhance plant resilience under salinity and extreme temperatures.
Tea anthracnose, caused by Colletotrichum camelliae, poses a serious threat to tea yield and quality. While certain transcription factors have been implicated in stress responses, the core transcriptional networks governing immunity in tea plants remain largely elusive. In this study, we identify CsWRKY33 as a central regulator that orchestrates immune responses and metabolic reprogramming during C. camelliae infection in tea plants. Transcriptome analysis revealed that CsWRKY33 is significantly induced upon fungal challenge and functions downstream of both pattern-triggered immunity (PTI) and effector-triggered immunity (ETI). Mechanistically, protein-promoter affinity experiments demonstrated that CsWRKY33 directly binds to W-box motifs in the promoters of key immune genes, including CsSERK2, CsMPK3, CsCDPK32, CsPR1B, CsWIN2 and CsRPM1. Dual-luciferase assays and functional validations further confirmed that CsWRKY33 activates its expression, forming a positive feedback loop that amplifies immune signalling. Concurrently, CsWRKY33 modulates primary metabolism by repressing photosynthetic genes such as CsRCA and enhances secondary metabolism by directly activating phenylpropanoid pathway genes such as CsPAL and CsCAD1. These coordinated transcriptional programmes reallocate energy and resources toward defence, thereby enhancing resistance. We propose a working model in which CsWRKY33 serves as a transcriptional hub linking immune activation with metabolic adjustment to facilitate efficient defence. This study provides new insights into WRKY-mediated immunity in perennial woody plants and highlights CsWRKY33 as a promising target for molecular breeding of anthracnose-resistant tea cultivars.
Substance use disorders (SUDs) pose a major global health burden, often worsened by comorbid depression and anxiety. Dysregulation of monoaminergic neurotransmission, especially dopamine, serotonin, and norepinephrine transporters, underlies the reinforcing and harmful effects of addictive drugs. Current pharmacotherapies targeting these transporters offer benefits but are limited by delayed onset, side effects, and modest efficacy against emotional and cognitive symptoms. Despite advances in structural biology, a unified framework integrating transporter structure with docking and molecular dynamics simulations remains lacking. Emerging evidence suggests that plant-derived metabolites and essential oil preparations may modulate monoamine transporters through both direct and indirect mechanisms. These includes allosteric effects, membrane interactions, ion channel modulation, and downstream signaling pathways; however, these effects require further validation. This review summarizes recent preclinical and clinical data on transporter-modulating metabolites from Hypericum perforatum L., Rhodiola rosea L., Withania somnifera, and some of the essential oils. It highlights mechanistic insights from structural biology and molecular pharmacology, suggesting that plant-derived metabolites and essential oil preparations may influence monoaminergic neurotransmission and stress-related pathways. Despite challenges in bioavailability, standardization, and clinical validation, these metabolites may offer polypharmacology for adjunctive, personalized SUDs interventions. Integrated approaches, merging structural modeling, enhanced delivery, and rigorous trials, are needed.
Neurodegenerative diseases are characterised by the progressive dysfunction of neurons, and memory impairment is one of their most debilitating clinical manifestations. The etiopathogenic mechanisms are multifactorial, such as protein misfolding, oxidative stress, and mitochondrial malfunction and synaptic degeneration along with neuroinflammation. As yet, therapists still focus mainly on symptomatic treatment and no agents are found to halt or reverse the decline of cognitive, indicating what is more needed is other kinds of neuroprotective strategy. In this context, the plant-derived phytochemicals stand out as promising candidates owing to their multi-targeted mode of action, favourable safety profile and long-standing use in traditional medicine systems. These bioactive compounds modulate oxidative stress, inflammatory signalling, neurotransmitter balance, apoptotic pathways and protein aggregation to elicit neuroprotection. The attention of the research community has also turned towards using Drosophila melanogaster as a model system for neurodegenerative-related studies due to its genetic tractability, accessible behavioural learning and memory tests, and the evolutionary conservation of potentially important biological pathways. This review consolidates recent evidence regarding plant-based neuroprotective strategies against memory impairment, with a specific focus on mechanistic mechanisms elucidated from Drosophila models of neurodegenerative diseases. Integrating findings across molecular, cellular and behavioural levels, the review illustrates the therapeutic promise of phytochemicals and reaffirms Drosophila as a valuable preclinical tool. It also addresses practical translational considerations, such as bioavailability, standardisation, and clinical validation, and sets forth future directions for effectiveness of plant-based interventions to facilitate improvements out in the real world.
Salvia hispanica L. (chia) seeds are recognized as a functional food rich in phenolic compounds, yet the mechanistic basis linking their chemical profile with antidiabetic activity remains insufficiently defined. To evaluate the antioxidant properties, α-glucosidase inhibition, antihyperglycemic and hypoglycemic responses of chia seed extracts of increasing polarity, and to characterize their major phenolics using UPLC-DAD-ESI-MS and molecular docking. n-hexane, dichloromethane, and methanolic extracts were assessed in normoglycemic Wistar rats using an oral sucrose tolerance test and an acute hypoglycemia model. In vitro inhibition of maltase, sucrase, and starch-degrading enzymes was quantified. Total phenolic content and antioxidant activity were determined by Folin-Ciocalteu, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. UPLC-DAD-ESI-MS tentatively identified phenolic constituents, and molecular docking was performed against α-glucosidase catalytic domains. The methanolic extract (MESh) exhibited the highest phenolic content and antioxidant activity, a phenolic profile dominated by rosmarinic acid, caffeic acid, and their hexosides, and the strongest antihyperglycemic effect, producing a glycemic curve comparable to acarbose. MESh moderately inhibited sucrase (61.5%), while the dichloromethane extracts moderately inhibited maltase (62.9%). Docking analyses revealed that rosmarinic acid and its hexoside displayed high binding affinities (-7.7 to - 8.1 kcal/mol) toward α-glucosidase targets, closely approaching those of acarbose (-7.9 to - 9.1 kcal/mol) and exceeding those of caffeic-acid derivatives, supporting their potential contribution to the in vitro and in vivo responses. Chia seed extracts exhibit complementary antidiabetic-related activities involving antioxidant effects, modulation of intestinal α-glucosidases, and attenuation of postprandial glycemic excursions. Rosmarinic-acid derivatives emerge as key contributors to these effects. These findings reinforce S. hispanica as a promising functional food for glycemic regulation.
In Arabidopsis, loss of transcription factors Calmodulin-Binding Transcription Activators 1/2/3 (CAMTA1/2/3) results in enhanced defense responses with increased salicylic acid (SA) and N-hydroxy-pipecolic acid (NHP) biosynthesis. This is achieved through modulating the expression of master immune transcription factors SAR Deficient 1 (SARD1) and Calmodulin-Binding Protein 60-like g (CBP60g). SARD4 is an NHP biosynthesis enzyme, the camta2/3 sard4 triple mutant remains dwarfed and autoimmune. To identify positive immune regulators of the SA and NHP pathways, a forward genetic screen was performed using the camta2/3 sard4 mutant. Here, we report on a loss-of-function allele of Late Upregulated in Response to Hyaloperonospora parasitica 1 (lurp1) that suppresses the autoimmunity of camta2/3 sard4. LURP1 transcripts are strongly induced by pathogens. Chromatin immunoprecipitation and expression analyses revealed that LURP1 expression is directly controlled by SARD1. LURP1 belongs to a small gene family. Knocking out LURP1 and its three close homologs compromises both flg22- and nlp20-induced resistance and SA accumulation, supporting their redundant roles in promoting plant defense. Interestingly, LURP1 interacts with the helper nucleotide-binding leucine-rich repeat receptor (hNLR) Activated Disease Resistance 1 (ADR1) family, suggestive of a model where LURP1 boosts pattern-triggered immunity and SA-mediated defense signaling through the ADR1s.
China's soybean supply is heavily dependent on international markets, making it an urgent task to enhance domestic self-sufficiency to ensure food security. Data from the national soil census indicate that China possesses approximately 500 million mu of saline-alkali soil resources, of which about 200 million mu have potential for agricultural development. Against the backdrop of tight arable land resources, developing new soybean varieties tolerant to saline-alkali conditions represents a strategic initiative to effectively utilize saline-alkali land, expand cultivation areas, and address the challenge of soil salinization. Rapid alkalinization factors (RALFs) are a class of plant small peptides that act as ligands, initiating downstream signaling by binding to plasma membrane receptor complexes, thereby coordinating plant growth, development, and stress responses. However, the specific molecular mechanisms by which RALF peptides mediate responses to saline-alkali stress in important crops such as soybean remain unclear. Through expression profiling analysis of the soybean RALF family, combined with transcriptome data under alkaline salt treatment, this study identified two homologs of Arabidopsis thaliana AtRALF34, designated GmRALF34a and GmRALF34b, which are predominantly expressed in roots and whose expression is significantly suppressed following alkaline salt treatment. Using gene editing technology, we generated Gmralf34ab double mutants, which exhibited enhanced sensitivity to alkaline salt stress. In contrast, no significant differences were observed between the mutant and wild type plants under neutral salt stress. Field phenotypic characterization further demonstrated that the mutants showed significant reductions in agronomic traits, including plant height, node number, and yield per plant. In conclusion, this study preliminarily reveals that GmRALF34s play an important role in soybean response to alkaline salt stress and adaptation to saline-alkaline environments, provides valuable genetic materials for further elucidating their molecular mechanisms and establishing a theoretical and material foundation for breeding salt-alkali tolerant soybean varieties. 我国的大豆(Glycine max)供给严重依赖国际市场，提升国内自给能力已成为保障粮食安全的紧迫任务。全国土壤普查数据显示，我国拥有约5亿亩盐渍土资源，其中约2亿亩具备农业开发潜力。在耕地资源紧张的背景下，培育耐盐碱大豆新品种，是实现盐碱地资源有效利用、扩大种植面积并应对土壤盐渍化挑战的战略性举措。快速碱化因子(rapid alkalinization factor，RALF)是一类作为配体的植物小肽，通过结合质膜受体复合物启动下游信号，协调植物生长发育与逆境响应。然而RALF小肽在大豆等重要作物中介导盐碱胁迫应答的具体分子机制尚不明确。本研究通过对大豆RALF家族进行表达模式分析，并结合碱性盐处理下的转录组数据，发现拟南芥(Arabidopsis thaliana)的AtRALF34在大豆中的两个同源基因GmRALF34a和GmRALF34b在根中优势表达，且其表达在碱性盐处理后受到显著抑制。利用基因编辑技术获得Gmralf34ab双突变体，该突变体表现为对碱性盐胁迫敏感性增强，相比之下，在中性盐胁迫下，突变体与野生型无显著差异。大田表型鉴定进一步表明，突变体在株高、节数和单株产量等农艺性状上均显著降低。综上，本研究初步揭示了GmRALF34s在大豆响应碱性盐胁迫及盐碱地适应性中扮演着重要角色，为深入解析其分子机制提供了重要遗传材料，并为大豆耐盐碱育种奠定了理论与材料基础。.
The increasing demand for sustainable agricultural practices has highlighted the potential of Plant Growth-Promoting Bacteria (PGPB) as eco-friendly tools to enhance crop productivity while minimizing environmental impact. Among PGPBs, members of the Actinomycetota phylum (formerly known as actinobacteria), and particularly Streptomyces violaceoruber, have emerged as promising candidates due to their ability to produce bioactive metabolites, promote plant growth, and modulate plant physiological responses. In this work, we investigated the effects of S. violaceoruber on tomato (Solanum lycopersicum) in vitro - grown seedlings using an integrated phenotypic, volatilomic, transcriptomic, and epigenetic approach. Seedlings were analyzed at seven (T1) and fifteen days (T2) post-inoculation with S. violaceoruber. Phenotypic assessment of inoculated seedlings revealed no significant alterations in shoot length or biomass, while a remarkable increase in seedling root diameter and the formation of aerial roots was observed. Transcriptomic analyses showed substantial transcriptional reprogramming, with a greater number of differentially expressed genes (DEGs) at T1, in particular involved in the regulation of biological processes, metabolic pathways, and responses to external stimuli, such as light. Co-expression network analysis of four root-associated bait genes further confirmed that these pathways are primary targets of S. violaceoruber effects. Epigenetic profiling of treated plant roots revealed an increase in global DNA methylation (5-methylcytosine) levels, along with a significant enrichment of histone post-translational modifications associated with permissive chromatin at the bait gene loci. Overall, S. violaceoruber inoculation induced notable molecular and developmental changes in tomato seedlings, reinforcing its potential as a sustainable biofertilizer. These findings provide new insights into PGPB-plant interactions and contribute to the development of environmentally-friendly strategies for crop improvement.
Hydrothermal carbonization (HTC) represents an innovative and sustainable chemical approach based on sub-critical water and useful to valorize waste biomass to develop a novel solid material with a huge potential for soil microbiota and plant productivity. Two hydrochar types were produced, under mild (180 °C, 10 Bar) and severe (215 °C, 20 Bar) HTC conditions (HC180 and HC215, respectively), by processing spinach, red chicory, and escarole wastes resulting from industrial horticultural production. These hydrochars were characterized for elemental composition and molecular composition via advanced techniques (13C CPMAS NMR and FT-ATIR) and compared with the values detected for initial biomass types. HTC progressively converted labile into carbon-dense, aromatic materials, with greater severity reducing nutrient availability. Solid hydrochars retained up to 2.82 g of water per g of material, thus representing a tool to make soil more resilient against the drought. Greenhouse pot experiments on baby-leaf lettuce revealed that HC180 strongly promoted plant shoot and root growth, matching conventional mineral fertilization, while HC215 showing only limited effects. 1H NMR metabolomics indicated that HC180 stimulated primary metabolism, increasing the level of sugars and amino acids, linked to energy production and stress responses, highlighting a biostimulant effect mediated by enhanced nutrient availability and soil-microbiome interactions. These findings demonstrate that mild HTC conditions produce safe hydrochars with plant biostimulant activity, characterized by a balanced composition of labile carbon and nitrogen as well as a spectrum of plant-available nutrients. This offers a tunable strategy for sustainable horticultural residue management to produce a product enhancing soil fertility and vitality.
Freshwater scarcity has increased interest in circular water-use strategies, including the reuse of treated wastewater (TWW) for crop irrigation. While TWW contains nutrients beneficial for plant growth, it may also carry contaminants such as toxic metals (TM), which can be taken up by plants and pose risks to both crop and food safety. This field study investigates the influence of nanoplastics (NP), an emerging pollutant commonly found in waters, on TM bioaccumulation and antioxidant compound production in lettuce (Lactuca sativa L.), one of the most widely cultivated and consumed leafy vegetables in Western countries. Lettuce plants were grown under greenhouse conditions and irrigated throughout the entire growth cycle with TWW, either alone or supplemented with NP, TM, or both. The results indicate that the interaction between NP and TM leads to a decreased TM bioaccumulation (24%, 50%, and 79% for Cr, Ni, and Cd, respectively) and reduced phytotoxicity, although with negative impact on plant growth (ca. 60% reduction in biomass production). To the best of our knowledge, this is the first study to investigate the effects of nanoplastics and co-existing contaminants present in treated wastewater on crops. These findings provide new insights into plant responses under irrigation with TWW and highlight potential implications for crop safety within the context of circular water use.
Neuropathic pain (NP) is a frequent sequel to peripheral nerve injury and maladaptive nervous system function. Daphnetin (DAP), a bioactive 7,8-dihydroxycoumarin derived from the traditional Chinese medical plant Daphne giraldii Nitsche, has potential anti-nociceptive effects and alleviated NP following intrathecal injection. However, the molecular target of DAP for this action remains unknown. This study aims to investigate the underlying mechanisms of DAP against NP. Chronic constriction injury (CCI) rat model and lipopolysaccharide (LPS)-treated HAPI microglial cells were established to evaluate the analgesic and anti-inflammatory effects and underlying mechanisms of DAP. Rat behavioral assessments included mechanical withdrawal threshold (MWT), thermal withdrawal latency (TWL), and gait analysis. The expression of mRNA was determined by qRT-PCR. Protein expression levels were detected by Western blot, immunofluorescence, and enzyme linked immunosorbent assay. Molecular docking, surface plasmon resonance, molecular dynamics simulation, and site-directed mutagenesis were employed to investigate the interaction between Kelch-like ECH-associated protein 1 (Keap1) and DAP. DAP treatment significantly increased MWT and TWT in CCI rats. It also improved gait parameters and attenuated spinal inflammation, neuronal damage, and hyperexcitability in CCI rats. DAP treatment markedly increased the expression of Nrf2 and its nuclear translocation in the rat spinal cord and HAPI cells. DAP treatment inhibited the activation of NLRP3 inflammasome and microglial M1 phenotype polarization, while promoting M2 phenotype polarization. Furthermore, the NLRP3 inhibitor MCC950 also alleviated hyperalgesia in CCI rats, inhibited NLRP3 inflammasome activation, suppressed microglial M1 polarization, and promotes M2 polarization, whereas the Nrf2 inhibitor ML385 exerted the opposite effect. It reversed the regulatory effect of DAP on spinal microglial polarization in the CCI rat spinal cord and LPS-induced HAPI cells. Molecular interaction studies further demonstrated that DAP binds to Tyr334 and Tyr572 residues in the Kelch domain of Keap1, thereby blocking Keap1/Nrf2 interaction to activate Nrf2. DAP treatment inhibits microglial M1 polarization and promotes M2 polarization via the Nrf2/NLRP3 inflammasome pathway, attenuates neuroinflammation and central sensitization, and finally achieves analgesic effects. These findings identify DAP as a novel Nrf2 activator with therapeutic potential for the management of NP.
Neuroinflammation, characterized by dysregulated activation of microglia, is a hallmark of Parkinson's disease (PD). Nevertheless, therapeutic strategies aimed at the mechanism of inflammation resolution remain limited. This study integrated PD clinical cohorts, MPTP-induced and miR-146a-induced mouse models, as well as LPS-stimulated and miR-146a-activated cell models. Combined with omics analysis, behavioral detection and molecular biology experiments, we systematically evaluated the anti-inflammatory protective effects of Wuzi Yanzong Pills (WYP). The active plant metabolites of WYP were identified using a combination of UHPLC-Q-Exactive-MS/MS, AP-SMALDI Orbitrap MSI, and pharmacokinetic analysis. In PD patients, WYP significantly improved motor dysfunction, inhibited pro-inflammatory cytokines, and elevated neurotransmitter levels. Exosomal miRNA sequencing analysis indicated that miR-146a-5p may serve as a biomarker for PD and is positively correlated with disease severity. Animal experiments further showed that WYP improved motor symptoms and neuroinflammation in MPTP- and miR-146a-induced PD mice models. A dual-luciferase reporter assay confirmed ubiquitin specific peptidase 3 (USP3) as a direct target gene of miR-146a-5p. In an LPS-activated BV2 microglial cell model, WYP intervention reduced the content of miR-146a-5p in cell-derived exosomes and mitigated their pro-inflammatory damaging effects on neuronal cells. Component analysis revealed that 16 plant metabolites in WYP can enter the bloodstream, among which 11 can cross into the brain. Notably, geniposidic acid, hyperoside, kaempferol, protocatechuic acid, and schisandrol A significantly suppressed the expression of pro-inflammatory factors in BV2 cells, suggesting that they may be the main active components underlying the anti-inflammatory effects of WYP. WYP improves PD by regulating the miR-146a-5p/USP3/NF-κB pathway. Meanwhile, the active plant metabolites of WYP have been identified. These findings provide experimental evidence for WYP as a potential therapeutic agent for PD.
Environmental pollution resulting from heavy metals constitutes a critical global issue. Remediation technologies offer potential solutions, particularly through the innovative use of endophytic microbes, either independently or in conjunction with plants. This solution is based on the ability of certain endophytic bacteria to produce metallophores, which are low-molecular-weight compounds capable of chelating various heavy metals. This study investigates ten bacterial endophytes isolated from the medicinal plant Galium aparine L. belonging to the Bacillus, Priestia, and Peribacillus genera. We tested different media to efficiently induce their production and assessed their ability to chelate various heavy metals, including highly toxic Pb2+, Cd2+ and Hg2+. Moreover, we examined in detail of their metallophore gene clusters, their organization, diversity and prevalence, by broad homology search. All strains exhibited moderate to high metallophore production ability, with few strains capable of chelating more metals than iron. Among them, Priestia sp. GS2 was identified as promising producer, reaching up to 60% SU, with binding activity also towards Co2+, Mn2+, Zn2+, Ni2+ or Cu2+. Also, Peribacillus frigoritolerans GR2 exhibits a remarkable ability to chelate Pb2+, Hg2+ and Cd2+. An in-depth analysis of the biosynthetic gene clusters and enzymes involved in metallophore biosynthesis revealed homologous clusters within previously deposited genomes, highlighting their distribution and potential evolutionary conservation. The strains demonstrated capacity for metallophore production and heavy metal chelation, which makes them promising candidates for the development of advanced microbial solutions. A genome-guided selection approach can guide the selection of strains for agricultural applications, where they enhance plant nutrient uptake, suppress soil pathogens, and support sustainable fertilization strategies beyond sequestering crucial metals. Apart from agriculture, purified metallophores can aid bioremediation and mobilization of heavy metals from various environments and matrices.