Bt (Bacillus thuringiensis) cotton exhibits effective resistance against lepidopteran insects; however, substantial declines in both the yield and quality of cotton plants are induced by low-temperature stress, concurrently decreasing Bt protein expression and insecticidal efficiency. As a primary fiber crop of significant industrial interest, Bt cotton requires stable agronomic performance under abiotic stress. This study employed a three-factor completely randomized experiment within a controlled-environment chamber to evaluate the effects of seven composite treatment groups (a water control and six exogenous regulators administered at 16°C) across three recovery sampling times (48, 72, and 96 h) on Bt toxin levels and nitrogen metabolic status in boll shells of two Bt cultivars. Results demonstrated that foliar application of 20 mmol/L CaCl₂ was the most effective in maintaining Bt protein stability during stress, resulting in a measured content numerically comparable to the optimal temperature control; chitosan (CTS) and salicylic acid (SA) exhibited partial mitigation effects.During the recovery phase (48–96 h), measured Bt protein content under 6-BA treatment recovered significantly compared with the water-sprayed control based on pooled data across growing seasons and cultivars (Tukey's HSD, P<0.05, 12 biological replicates).Partial least squares path modeling (PLS-PM) revealed that at the end of chilling stress (48 h), soluble protein content (0.360**) and proteolytic enzyme activity (−0.370*) exerted significant direct positive and negative effects on Bt protein content, respectively. During the recovery phase (96 h), proteolytic enzyme activity was the primary direct inhibitory factor (-0.599**), whereas transamination (48 h: 0.918**; 96 h: 0.992**) and free amino acid content (48 h: 0.574**; 96 h: 0.942**) indirectly regulated Bt protein accumulation by altering soluble protein synthesis. This study reveals that exogenous regulators facilitate the shift of boll shell nitrogen metabolism from a ‘degradation-dominant’ to a ‘synthesis-dominant’ mode by enhancing protein synthesis and suppressing proteolysis.
Low risky establishment of Miscanthus × giganteus requires optimal management strategies that ensure stable nutrient availability and fast, uniform early growth. This study evaluated the effects of different amendments—organic materials, biomass combustion ash, and urea—on seasonal soil chemical dynamics and first‑year biomass production in an outdoor pot experiment. Seasonal monitoring revealed pronounced shifts in nitrogen availability, with high early‑spring values, a decline toward late summer, and partial recovery in autumn. Whereas total carbon decreased until mid‑May, increased through late summer, and declined again in autumn. The highest average nitrogen mineralization rate occurred in mid‑spring, followed by a gradual decrease, while immobilization prevailed toward the end of the season. A concurrent reduction in the C:N ratio was also observed. No significant differences among amendments were detected in CO₂ emissions, whereas soil moisture was lowest in the control and highest in the biomass ash treatment. Initial rhizome traits did not significantly affect biomass production, which was most strongly associated with shoot number and maximum plant height. The highest biomass yield was obtained with cattle manure, and the lowest with biomass combustion ash application. Variance partitioning confirmed the substantial role of amendments in early biomass production, with rhizome traits contributing only marginally. Amendments also markedly altered biomass allocation: willow compost and farm‑residue compost increased the shoot proportion, whereas mineral amendments enhanced the share of rhizomes and roots
Renewable biomacromolecules, such as plant proteins, are appealing materials for producing sustainable, high-performance, biobased materials because of their biodegradability, structural versatility, wide availability, and extensive chemical functionality. Awareness of environmental issues associated with petroleum-based plastics and the increasing need for solutions in the field of circular bioeconomy have intensified interest in plant protein-based materials for packaging, biomedical, agricultural, textile, and advanced engineering applications. However, the use of native plant proteins in industry is restricted due to their moisture sensitivity, low mechanical strength, limited thermoplasticity, and batch-to-batch variability. Thus, advanced functionalization strategies are required to structure and optimize the structure–property relationships and, consequently, to improve the material performance. This article critically summarizes the latest developments in the extraction and sustainable processing of plant proteins from various sources, including soy, wheat, maize, pea, rice, potato, sunflower, and rape. This review also discusses the functionalization of plant proteins. Special focus is placed on the molecular architecture, amino acid composition, intermolecular interactions, and hierarchical assembly mechanisms of physicochemical and functional behaviors. The physical, chemical, enzymatic, and nanomodification methods are compared and discussed, along with their effects on mechanical properties, thermal stability, rheology, barrier properties, biodegradability, and environmental responsiveness. This review also emphasizes the formulation of multifunctional plant protein-based materials, such as films, coatings, hydrogels, aerogels, electrospun fibers, nanocomposites, adhesives, and smart, responsive systems. The development and potential of emerging technologies, such as nanotechnology, additive manufacturing, and AI-assisted biomaterial design, are explored as potential solutions for next-generation sustainable materials. Finally, the significant hurdles to the commercialization of plant protein-based materials, such as feedstock variability, processing scalability, environmental durability, regulatory issues, and techno-economic challenges, are critically analyzed. This review offers a holistic, mechanistic, and translational approach to the future of plant protein biomaterials for scalable, multifunctional, and environmentally friendly applications.
Straw-derived humic substances (HS) represent a bio-based material produced from agricultural residues, with well-defined chemical properties suitable for remediating heavy metal-contaminated soils. However, the mechanisms through which HS drive remediation, particularly by regulating microbial metabolic networks, remain poorly understood. Here, we conducted a soil column experiment to investigate the effects of straw-derived HS on the remediation of soils co-contaminated with cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn). Characterization revealed that the straw-derived HS possessed abundant carbonyl (31.0%) and carboxyl (12.2%) groups, a high humification ratio (93.3%), and a low molecular weight (1838 Da). HS application significantly reduced the total contents of heavy metals in soil by 10.5–14.9% and their bioavailability by 22.2–31.5%. It also improved soil structure, nutrient availability, and carbon-nitrogen cycling, thereby facilitating metal complexation and supporting beneficial microbial reorganization. Together, these changes promoted complementary bacterial-fungal interactions that enhanced system stability under metal stress. Metabolomic analysis revealed that HS activated energy metabolism and organic acid synthesis pathways, establishing a previously unreported microbe-metabolite network centered on Pedobacter and 2-decyl-3-hydroxy-pentanedioic acid. These findings provide a scientific basis for valorizing crop straw into HS as a sustainable industrial product for environmental soil management.
The saccharide content is one of the most important indicators for the quality evaluation of the Shihu herb (Dendrobium nobile Lindl.). To fully understand saccharide production in this medicinal plant, an integrated metabolic and transcriptomic method was employed for analysis. The contents of total saccharides, non-phosphorylated saccharides, and hexose saccharides were mainly induced by low temperature and drought, while they were suppressed by dark and ultraviolet stresses. There were 44 saccharides and saccharide derivatives showed a significant stress response in Dendrobium nobile. Following genome-wide association and protein-protein interaction, a total of 182 genes were identified to participate in the core regulation of saccharide production. Based on these differentially accumulated saccharides and differentially expressed genes, a saccharide biosynthesis map was then constructed. From this map, the saccharide production was mainly improved by enhancing carbon fixation and return. Finally, a complete regulatory network from environmental stresses to saccharide accumulation was further deduced, including stress recognition (DnHSP70s, DnUVR8, and DnCOR413PM2), signal transduction (DnTMK1, DnCDKB1–1, and DnHFLX), transcriptional regulation (DnPTI6, DnSRM1L, DnNAC3), genetic information processing (DnRPLs and DnRPSs), post-translational regulation (DnDNAJs), photosynthesis (DnMPUHL1, DnPSA3C, and DnRAF1C), carbon fixation (DnRBCS512 and DnPPCK), return of down-stream metabolites (DnPEELs, DnCHNs, DnGEBGUs, and DnBAMs), saccharide biosynthesis (DnA1E, DnAATPSs, and DnSUS1), and saccharide transport (DnERD6L16, DnWRPC, and DnPP2A2). These findings provide valuable insights into improving saccharide production through molecular breeding and wild like cultivation. They will be beneficial for saccharide based application of Dendrobium nobile in the healthcare and pharmaceutical industry.
Cottonseed meal (CM) with high production has high crude protein (more than 40 %) and a balanced amino acid profile (similarity to FAO/WHO recommendations), therefore it is a promising protein source to develop active peptides. However, CM is often seen as a low-value waste leading to environmental pollution due to high levels of free gossypol, the dephenolized cottonseed meal (DCM) might be a better raw material to obtain active peptides. In this study, strains were screened by high capacity of hydrolyzing DCM gossypol, and strain combinations were selected for hydrolyzing cottonseed proteins. A Box-Behnken design was used to optimize fermentation parameters to maximize the yield of soluble peptides, and antioxidant peptides with high absorption were obtained by LC-MS/MS, Pre-AnOxPs database and admetSAR 3.0. The antioxidant activities of these peptides were validated by cell experiments. Results showed that the strongest free-degrading strain was Bacillus velezensis WY122 (B.velezensis WY122), and the optimal combination of degrading cottonseed proteins was B.velezensis WY122 and Lactiplantibacillus plantarum 01002 (L.Plantarum 01002). Under optimal fermentation conditions, a maximum soluble peptide yield of 211.855 mg g⁻¹ of fermented DCM was obtained, and higher DPPH and Fe²⁺-chelating capacities were found. The gossypol in DCM was degraded into salicylic acid and catechol. The 42 peptides with antioxidant potential were identified, and the hydrophobic AA residues (Phe, Leu, Pro, Val, Ala and Gly) accounted for a large proportion among them. These five peptides (VSLLLPR, FLVR, FLLR, VVDLLR, and YLDDLAELR) were demonstrated to have antioxidant activity, and VSLLLPR had the strongest antioxidant capacity. Overall, a new strain was found, and a novel method for developing active peptides from DCM was established, which provided theoretical basis for developing cottonseed peptides as functional materials. Fermentation provided an effective way for converting low-value agricultural byproducts into high value-added products.
Erianin is a bibenzyl compound with remarkable anti-tumor activity, abundantly present in the medicinal orchid Dendrobium chrysotoxum Lindl. However, the precise enzymatic steps governing erianin biosynthesis in D. chrysotoxum remain partially uncharacterized, limiting its potential for medicinal development and utilization. In this study, the accumulation dynamics of erianin were systematically tracked across different developmental stages of D. chrysotoxum stems. An inverse accumulation pattern was observed between erianin and another major bibenzyl, gigantol, during stem development. Metabolomic analysis of stems at three representative developmental stages led to the proposal of a six-step biosynthetic pathway for erianin, in which four steps are catalyzed by O-methyltransferases (OMTs), underscoring their central role. A comparative transcriptomic analysis identified 17 candidate OMT genes, three of which were confirmed to have the expected function through in vitro and in vivo enzyme activity assays and tobacco transient transformation system validation experiments. Furthermore, the three identified DchOMTs exhibit distinct substrate specificities, with each one catalyzing a different step in the erianin biosynthetic pathway. Molecular docking further supported the stable binding affinity between each OMT and its corresponding substrate. This study elucidates key molecular mechanisms involved in erianin biosynthesis and provides a genetic explanation for its high accumulation in D. chrysotoxum. The findings offer valuable targets and gene resources for the metabolic engineering and synthetic biology of bibenzyl-derived anticancer natural products.
Economic fruit trees are important cash crops, and their fruits serve as key raw materials for processing dried fruits, fruit wine, juice, jam and other value-added products. Standardized orchard mechanization guarantees stable and high-quality raw material production by realizing core procedures including fertilization, plant protection, pruning and harvesting through machinery–crop–environment coupling interactions. The Discrete Element Method (DEM) is an effective numerical approach for discontinuous medium analysis, and it is widely adopted to reveal mechanistic principles of orchard operations and optimize equipment parameters, with modeling accuracy directly affecting simulation reliability. Existing DEM reviews primarily focus on general agricultural mechanization, whereas few studies systematically summarize DEM applications across the full industrial chain of fruit crops, spanning field cultivation, mechanized operation and post-harvest processing. This review introduces the fundamental theories and core modeling techniques of DEM, focusing on contact model selection, particle modeling, and parameter calibration for typical orchard materials such as soil, fertilizer, tree branches, fresh fruits and processing residues. This work comprehensively reviews DEM applications in field operations including seedling planting, fertilization, plant protection, pruning and mechanical harvesting, and further extends the analysis to post-harvest processing scenarios such as fruit grading, conveying, separation, crushing, drying and deep processing. The main limitations of current orchard-oriented DEM simulations are concluded, including insufficient model accuracy and universality, low computational efficiency and inadequate multi-physics coupling performance. Future research directions and technical optimization strategies are accordingly proposed, providing theoretical references for the standardized and industrialized production of economic fruit crops.
Accurate thermal conductivity estimation is critical for optimizing fuel atomization and combustion efficiency in modern engines. This study introduces a high-fidelity data-driven framework for predicting the thermal conductivity of biodiesel–additive blends across extensive thermodynamic ranges, including 249–523 K and pressures up to 42.75 MPa. Utilizing a comprehensive repository of 2811 experimental data points, four computational paradigms, including least-squares boosting (LSBT), radial basis function neural networks (RBFNN), adaptive neuro-fuzzy inference systems (ANFIS), and bagged trees (BT), were developed using a unified seven-dimensional input vector. Bayesian optimization was employed for hyperparameter tuning to ensure a robust bias–variance trade-off. Statistical evaluations revealed that the LSBT model provided the best predictive performance, achieving a validation mean absolute percentage error (MAPE) of 0.56%, a relative root mean squared error (RRMSE) of 0.72%, and an R2 of 99.52%. Robustness was verified through 5-fold cross-validation, exhibiting a negligible generalization gap, while William’s plot analysis confirmed that 99.08% of the data resided within the valid applicability domain. Notably, the developed models reproduced physically consistent thermal conductivity trends, particularly the non-monotonic relationship with molecular chain length and an apparent transition around 8 carbons, which may reflect changes in molecular packing, polarity, and dispersion-dominated interactions as chain length increases. Furthermore, SHAP-based interpretability analysis identified temperature as the most critical driver while distinguishing between associative alcohol effects and van der Waals interactions in alkanes. This optimized framework serves as an accurate and interpretable data-driven surrogate for rapid thermophysical property estimation, reducing experimental effort and supporting large-scale computational tasks and industrial combustion modeling within the investigated domain.
Scutellaria baicalensis Georgi is a perennial herb belonging to the family Labiatae. Its roots are rich in flavonoids and possess heat-clearing, dampness-drying, fire-purging, and detoxifying properties. Under natural conditions, purple petals predominate in this species, whereas purple-red and white variants are rare. Previous transcriptomic and metabolomic analyses of flowers with different colors have mapped the anthocyanin biosynthetic pathway in S. baicalensis and identified differentially expressed candidate genes. However, the transcriptional regulation of anthocyanin structural genes remains unclear, and the SbR2R3-MYB gene family has not yet been systematically characterized. To address this knowledge gap, a genome-wide analysis of the SbR2R3-MYB family was conducted, with a particular focus on candidate genes involved in anthocyanin biosynthesis. A total of 55 members were identified and characterized based on their physicochemical properties, phylogenetic relationships, gene structures, sequence conservation, chromosomal distributions, and collinearity. Bioinformatic analyses identified SbMYB306 and SbbHLH96 as key regulatory genes and SbANS and SbDFR as anthocyanin-related structural genes for further investigation. Multiple assays, including yeast one-hybrid (Y1H), yeast two-hybrid (Y2H), dual-luciferase reporter, bimolecular fluorescence complementation (BiFC), and β-glucuronidase (GUS) activity assays, demonstrated that SbMYB306 and the SbMYB306–SbbHLH96 complex regulate key enzyme-encoding genes involved in flower color formation. Heterologous overexpression of SbMYB306 in tobacco enhanced petal pigmentation and increased anthocyanin accumulation. These findings establish SbMYB306 as a positive regulator of anthocyanin biosynthesis, support the evolutionary conservation of the anthocyanin biosynthetic pathway, and provide a theoretical basis for further investigation of flower color formation in S. baicalensis.
Panax notoginseng (P. notoginseng) is a valuable medicinal herb. Yunnan Province in China is the primary producing region, which is characterized by intensive mining activities and high geochemically background levels of metal(loid)s like arsenic (As), cadmium (Cd) and lead (Pb). However, whether and how much metal(loid)s can be accumulated and enter gastrointestinal tracts to induce health risk are still elusive. Therefore, paired P. notoginseng and soil samples (n = 140) were collected to investigate the transfer and risk of metal(loid)s within the soil-P. notoginseng-human continuum. The results showed that 45%, 15% and 9.29% of soil samples were polluted by As (11.9–231 mg kg–1), Cd (0.06–1.22 mg kg–1) and Pb (22–98.8 mg kg–1) (Pollution index, Pi>1). Soil Cd showed higher availability (18.5%) than Pb (4.44%) and As (3.86%). After uptake by P. notoginseng, Cd was mainly accumulated in taproots, while As and Pb were translocated from taproot to leaf (TF=0.69 vs. 5.35 and 2.84). Besides, P. notoginseng showed higher bioaccumulation factor (BAF) for Cd than for As and Pb (BAF=1.07–6.38 vs. 0.01–0.09 and 0.03–0.19). As such, 23.6% of taproot Cd and 16.3–62.1% of leaf As/Pb contents failed the food safety standards (0.3, 1.5 and 2 mg kg–1). In taproots, metal(loid) bioaccessibility was higher in gastric phase (GP) than gastrointestinal phase (GI) (58.8–77.2% vs. 26.9–46%) due to metal(loid) precipitation under alkaline environment in intestinal phase (IP) (pH=7.0–8.0). Although total content-based assessment showed potential health risk (3.57% target hazard quotient THQAs≥1 and 6.25% THQCd≥1) in taproots, bioaccessible content-based assessment suggested negligible-to-low concern risk for adults. These findings highlight the necessity of incorporating bioaccessible fraction into risk assessment. The bioaccessible metal(loid)-based assay can provide more exposure-relevant risk predictions and informed consumption decisions.
Gardenia jasminoides fruits are commercially utilized as a source of geniposide, the precursor of the cross-linking agent genipin, but the tissue-specific accumulation of bioactive flavonoids and their underlying biosynthetic mechanisms remain poorly understood. Here, we integrated antioxidant profiling, targeted metabolomics, gene expression analysis, and molecular docking to investigate fruit-specific flavones across five tissues of G. jasminoides. Among five tissues, the fruit exhibited the highest antioxidant activity. Targeted metabolomics of 2737 compounds revealed that the fruit's superior activity arises from its specific flavonoid composition, rather than total abundance. Notably, fruit-accumulating flavones were biosynthetically traced to flavone synthase II (FNSII). Two fruit-preferential FNSII isoforms were identified and showed endoplasmic reticulum localization. As a complementary computational prediction, molecular docking suggested that the fruit-specific flavone 3,6,2′,3′-tetramethoxyflavone, along with the aglycone of apigenosylide B, may binds to the Kelch-like ECH-associated protein 1 (KEAP1) domain with predicted affinity comparable to the known binder luteolin. This study provides the first evidence that flavonoid composition, rather than total abundance, governs the bioactivity of G. jasminoides fruits, and identifies 3,6,2′,3′-tetramethoxyflavone as a previously unreported candidate with predicted KEAP1-targeting candidate. These findings establish a chemical and molecular basis for the rational development of G. jasminoides fruits as a source of value-added industrial raw materials with defined quality markers.
High electrical conductivity is essential for electromagnetic interference (EMI) shielding, whereas efficient interfacial evaporation relies on highly porous architectures with low thermal conductivity. These conflicting requirements create an intrinsic trade-off, making it challenging for carbon-based foams to simultaneously achieve both functionalities. Herein, we overcome this intrinsic trade-off by engineering a bamboo-derived nanoflake-bridged carbon foam that preserves the intrinsic gradient hierarchical architecture of natural bamboo, where cellulose-derived nanosheets interconnect the microporous carbon framework to establish continuous conductive pathways without compromising rapid water transport. This synergistic structural design simultaneously enhances electromagnetic wave attenuation and interfacial evaporation, enabling efficient EMI shielding, high desalination efficiency, and robust salt-resistant operation. The resulting carbonized bamboo sponge (CBS) exhibits a high electrical conductivity of 40.65 S m−1 and an EMI shielding effectiveness of 34.04 dB in the X-band. Meanwhile, the continuous heat-generation pathways afford superior Joule heating performance, allowing the temperature to reach 103 °C at an ultralow voltage of 2.5 V. Beyond electromagnetic and thermal functions, the CBS uniquely integrates high-efficiency interface evaporation performance (7.65 kg m−2 h−1). This work establishes a sustainable design strategy for multifunctional carbon sponges, offering broad prospects in EMI mitigation and energy-water nexus technologies.
Platycodon grandiflorus is widely recognized as an industrial crop due to the high value of its bioactive polysaccharides (PGP). Despite its traditional use for pulmonary and gastrointestinal health, the underlying mechanisms remain poorly defined. This study investigated whether PGP intervention alleviates cigarette smoke (CS)-induced COPD in mice by restoring intestinal barrier integrity and reducing the translocation of lipopolysaccharide-carrying bacterial extracellular vesicles (LPS-bEVs) to the lung. Our results showed that a 7-week intervention with PGP significantly ameliorated CS-induced COPD symptoms, with the high-dose group exhibiting the most pronounced effects. PGP supplementation effectively mitigated body weight loss, pulmonary damage, intestinal shortening, and inflammatory cell infiltration. Notably, COPD-induced chronic mild hypoxia led to intestinal tissue hypoxia, which was alleviated by PGP through suppression of pathological glycolysis. This, in turn, restored the balance of cellular renewal and differentiation, improved intestinal barrier integrity, and limited the translocation of gut microbiota and their metabolites. In the lungs, PGP inhibited the activation of the non-canonical inflammasome pathway (Caspase11/GSDMD) triggered by LPS-bEVs, thereby attenuating pulmonary inflammation and preventing further injury. These findings establish PGP as a bioactive industrial crop component that regulates the gut-lung axis via dual mechanisms involving intestinal barrier protection and pulmonary pyroptosis suppression. This work provides a mechanistic foundation for developing PGP-based functional ingredients for respiratory health management, highlighting the value of P. grandiflorus beyond traditional applications toward evidence-based industrial product development.
Cadmium (Cd) contamination inhibits plant growth and threatens the safe production of crops, but the root-mediated regulatory mechanisms that simultaneously enhance Cd tolerance and reduce tissue Cd accumulation remain incompletely understood. This study aimed to determine the function and upstream transcriptional regulation of the tobacco expansin-like gene NtEXLB1 under Cd stress. NtEXLB1-overexpression and CRISPR/Cas9 knockout tobacco lines were analysed using yeast growth assays, physiological measurements, root histology, RNA-seq, yeast one-hybrid assays, and dual-luciferase reporter assays. NtEXLB1 was strongly induced by Cd stress, and its heterologous expression improved yeast growth under Cd treatment. In tobacco, NtEXLB1 overexpression alleviated Cd-induced inhibition of root growth and biomass and was associated with reduced Cd concentrations in roots, stems, and leaves, whereas knockout lines exhibited the opposite phenotypes. NtEXLB1 overexpression also maintained root tissue integrity, reduced ROS and malondialdehyde accumulation, and enhanced catalase and peroxidase activities. Transcriptomic analysis showed that NtEXLB1-associated genes were enriched in oxidative-stress responses, ROS metabolism, cell-wall remodelling, and root development. NtNAC50 directly bound to the NtEXLB1 promoter and activated its transcription. Collectively, these results identify an NtNAC50–NtEXLB1 regulatory module that contributes to Cd tolerance by strengthening root antioxidant defence and maintaining root structural integrity and is associated with altered Cd accumulation in tobacco tissues. NtEXLB1 therefore represents a potential target for improving Cd tolerance and reducing Cd accumulation in crops.
Essential oil production is increasingly framed not only as an extraction challenge but as a circular biorefinery problem in which essential oil, hydrosol, process water, and solid residues should be jointly valorized. This systematic review synthesized evidence on production technologies and sustainability performance using a PRISMA 2020-informed design. Searches structured for Scopus and Web of Science logic and validated across publisher platforms identified 40 eligible studies published between 2021 and 2026. Six technology families were mapped, with microwave-assisted distillation and by-product or residue valorization dominating the field. Most studies reported yield and compositional quality gains, whereas far fewer quantified environmental burdens or economic feasibility. The evidence shows that device-level intensification alone rarely proves circularity; stronger sustainability claims emerged when intensified extraction was combined with hydrosol use, residue valorization, life cycle assessment, or techno-economic analysis. However, the literature remains constrained by heterogeneous metrics, laboratory-scale designs, and limited system-boundary reporting. Overall, the review finds that the most credible pathway toward sustainable essential oil production is not a single superior extractor, but an integrated process design coupling efficient recovery with verified co-product cascading and system-level evaluation.
Upland cotton (Gossypium hirsutum), the world's dominant natural fiber crop, suffers severe yield and quality losses from Verticillium wilt. This highly destructive vascular disease, caused by the soilborne hemibiotrophic fungus Verticillium dahliae, poses a persistent threat to global cotton production. Nutrient transporters are increasingly recognized as candidate targets for innovative disease management, yet their functional roles and involvement in pathogenicity remain largely unexplored for non-canonical functions in V. dahliae. Here, we identified and functionally characterized VdIPT1 encoding a putative inositol transporter in V. dahliae. Deletion of VdIPT1 (ΔVdIPT1) unexpectedly promoted mycelial proliferation, conidiation capacity, and carbon/nitrogen assimilation, leading to enhanced virulence towards cotton seedlings. In agreement with fungal knockout phenotypes, both transient Host-Induced Gene Silencing (HIGS) and stable expression of VdIPT1-RNAi constructs in cotton effectively silenced fungal VdIPT1 but compromised host defense rather than conferring disease resistance, resulting in exacerbated disease symptoms and elevated fungal biomass accumulation. Transcriptome profiling demonstrated that the impaired resistance in VdIPT1-RNAi cotton was tightly linked to the transcriptional repression of core defense modules, particularly phenylpropanoid biosynthesis and plant–pathogen interaction signaling cascades. Our findings establish this putative inositol transporter VdIPT1 as a negative modulator of fungal virulence. Critically, silencing VdIPT1 is not a feasible HIGS-based resistance strategy. This work provides an important cautionary example, emphasizing that target genes must undergo thorough functional validation prior to their application in RNAi-mediated crop disease resistance breeding.
Green and efficient valorization of lignocellulosic biomass is pivotal to realizing sustainable biorefineries for a circular bioeconomy. Herein, an alkaline amino acid deep eutectic solvent (DES) pretreatment coupled with stepwise enzymatic hydrolysis was developed to improve selective delignification and facilitate co-production of xylo-oligosaccharides (XOS) and glucose. The aqueous choline hydroxide-arginine (ChOH-arginine) system enabled selective lignin removal while preserving hemicellulose for subsequent XOS production. Under mild conditions (80 °C, 30 min), a delignification rate of 84.25% was achieved, along with high retention of xylan (>71%) and cellulose (>86%). Density functional theory (DFT) calculations were performed to elucidate the underlying mechanism of lignin dissolution. Furthermore, the two-step xylanase-cellulase hydrolysis strategy delivered excellent XOS production performance. Critically, the pretreatment selectively cleaved lignin-carbohydrate ester linkages, suppressed lignin condensation, and retained a high β-O-4 linkage content, which are beneficial for the downstream valorization of lignin. Overall, this work proposes an integrated and sustainable strategy for efficient bioconversion of corn stover.
Circular RNAs (circRNAs) are critical regulators of plant growth, development, and stress adaptation; however, their roles in the biosynthesis of flavonoids, a class of pharmaceutically valuable secondary metabolites, remain poorly understood. Low flavonoid content in Ginkgo biloba leaves severely restricts its medicinal and industrial applications. Exogenous hormones effectively promote flavonoid accumulation, but the underlying circRNA mediated competitive endogenous RNA (ceRNA) regulatory mechanisms remain unclear. In this study, we treated G. biloba leaves with 24-epibrassinolide (EBR) and ethephon (ETH) and identified their optimal functional concentrations: 0.8 mg/L EBR increased quercetin and kaempferol accumulation, while 200 mg/L ETH elevated isorhamnetin content. Whole transcriptome sequencing identified 45 and 47 differentially expressed circRNAs (DECs), 135 and 289 differentially expressed miRNAs (DEMs), and 2232 and 2719 differentially expressed mRNAs (DEGs) under EBR and ETH, respectively. GO and KEGG analyses indicated that circRNAs host genes were consistently enriched in protein homeostasis pathways, indicating a core role of circRNAs in hormone induced stress responses. At the mRNA level, EBR primarily affected phenylpropanoid metabolism, whereas ETH modulated photosynthesis and stress responses. At the miRNA level, ETH uniquely enriched autophagy and DNA repair pathways, consistent with its robust global stress response signature. EBR significantly upregulated the flavonol synthesis genes GbFLS5, GbFLS6, and GbF3’H4, along with GbMYB8. Conversely, ETH induced the methylated flavonoid synthesis gene GbOMT and GbbHLH144. By integrating two independent methods, we further constructed putative ceRNA networks and screened key regulatory pairs involved in flavonoid biosynthesis, such as chr8_part1:429472903|429562118-novel_miR_202/448-GbbHLH6. Given that the functions of these ceRNA modules are preliminarily predicted, further experimental validation is required. This study reveals the divergent circRNA-miRNA-mRNA regulatory mechanisms underlying EBR and ETH induced flavonoid accumulation, providing promising targets for molecular breeding and industrial utilization of high flavonoid yielding G. biloba resources.
