Search Results (529)

In this study, potential fungicides were prepared following the principles of green chemistry. The compounds were synthesized in deep eutectic solvents as an alternative medium and compared with syntheses in traditional solvents such as ethanol. The efficiency of the reaction was improved by ultrasonic synthesis in both eutectic solvents and ethanol, resulting in higher yields while reducing reaction energy and time. For the first time, deep eutectic solvents (DES) were used for quaternisation reactions, with choline chloride as a hydrogen bond acceptor and urea, glycerol, malic acid, malonic acid, and levulinic acid as donors. DES, composed of biodegradable, non-toxic, and renewable components, represented a greener alternative to conventional solvents. However, reactions in DES by the conventional method generally resulted in lower yields, probably due to solubility and viscosity limitations inherent in the eutectic medium. The combination of ultrasound and deep eutectic solvents proved to be a good alternative to organic solvents for the quaternisation reaction, as higher yields were achieved in a shorter time compared to conventional methods. The antifungal activity of all 18 synthesized compounds was tested. The compounds exhibited significant antifungal activity against all four pathogens, with varying levels of mycelial growth inhibition. B. cinerea was the most sensitive species (up to 70.7% inhibition), while F. culmorum was the least sensitive (≤32%). Full article

Botrytis cinerea, the causative agent of gray mold, is a major fungal pathogen affecting a wide range of economically important crops. To identify sustainable alternatives to chemical fungicides, this study characterized the biocontrol potential of two bacterial strains, Serratia quinivorans NFX21 and Pseudomonas thivervalensis NFX104, through genomic analysis and functional assays targeting key stages of fungal growth and plant infection. The NFX21 and NFX104 strains significantly inhibited B. cinerea mycelial growth (~35%) and strongly suppressed conidial germination with performances comparable to the reference biocontrol strain Bacillus amyloliquefaciens QST 713. In tomato detached-leaf and whole-plant pot assays, application of NFX21 and NFX104 significantly reduced gray mold incidence and lesion severity relative to nontreated infected plants (53–64%, detached leaves; 12–13%, whole-plant assays), achieving disease control levels similar to those obtained with the commercial biofungicide Serenade ASO^®. Whole-genome sequencing allowed the taxonomic assignment of the NFX strains and revealed a rich repertoire of biosynthetic gene clusters and antifungal determinants. The NFX21 genome contained genes associated with N-acyl-homoserine lactone-mediated quorum-sensing and production of lipopeptides, siderophores, and extracellular lytic enzymes. The NFX104 genome harbored clusters involved in the biosynthesis of multiple siderophores, 2,4-diacetylphloroglucinol and hydrogen cyanide. Moreover, both the NFX21 and NFX104 genomes contained additional low-homology clusters that potentially encode for novel unexplored metabolites. Collectively, these results support the translational potential of NFX21 and NFX104 as biocontrol candidates for sustainable, integrated management of gray mold caused by B. cinerea. Full article

Finding safe and efficient plant protection measures is one of the major challenges in horticulture. This study evaluated the biological effects of the Thymus vulgaris essential oil and of Bacillus halotolerans and B. velezensis bacterial mixture on strawberry growth and fruit quality properties, as well as on Botrytis cinerea severity. The experiment was conducted in a high-tunnel greenhouse with the strawberry cv. Sonsation. Treatments: (1) Control—untreated; (2) Bacteria—with Bacillus halotolerans and B. velezensis, four times during flowering; (3) Thyme I—T. vulgaris essential oil (EO), four times during flowering; (4) Thyme II—T. vulgaris EO, four applications supplemented by three additional applications during fruit ripening; (5) Biofungicide I—Bacillus subtilis QST 713, four times supplemented by three applications; (6) Biofungicide II—Clonostachys rosea J1446, four times during flowering. In the first year (2023), the highest total yield was observed in Thyme II, and in the second year (2024), the highest total yield was observed in Thyme I. The results did not reveal any visual phytotoxic effect on plant leaves. The average fruit diameter increased from 28 mm up to 31 mm in 2023 and from 35 mm to 39 mm in 2024. The average soluble solids content increased from 9.4 to 11.4 °Brix in 2023 and from 7.2 to 7.7 °Brix in 2024. The highest ascorbic acid content in 2023 was observed in Biofungicide II and Biofungicide I treatments, respectively, 79.9 mg % and 75.4 mg %. Similarly, in 2024, the highest ascorbic acid content was observed in Bacteria, Biofungicide I, and Biofungicide II treatments—39.3–40.2 mg %. In vitro, the lowest B. cinerea severity on strawberry leaves in 2023 was recorded in Thyme I and Thyme II treatments (~6–7%), while in 2024, the severity in these treatments was higher −20–22%. Thyme treatment showed a stable reduction in B. cinerea on leaves in vitro over both years. Full article

The antimicrobial activity of multicomponent, magnetron-sputtered glass coatings was evaluated against phytopathogenic fungi (Botrytis cinerea, Fusarium oxysporum, Cladosporium fulvum, Alternaria solani) and the chromista Phytophthora infestans, with Aspergillus fumigatus included as a model opportunistic pathogen. Fourteen Cu-based multicomponent coatings were deposited on glass using multi-alloy targets composed of Sn, Zn, Al, Ni, Fe, Ti, Mn, Nb, or Co in two high-transmittance variants (≥85% and ≥88%). Antimicrobial activity was assessed in two assays: (A) spore survival after 24–72 h contact, and (B) hyphal growth over 7 days following coating exposure under light and dark conditions. Spore viability decreased after incubation on high-Cu coatings, which showed inhibition for most strains, particularly B. cinerea, F. oxysporum, and P. infestans. The effects on spore germination were independent of the direct transmittance value of the coated glass. Hyphal growth was generally less affected by a high Cu content for most strains. Hyphal growth of F. oxysporum, C. fulvum, A. solani and B. cinerea was reduced by up to 30% on selected multicomponent coatings. For most strains, hyphal growth showed no inhibition after light incubation on coatings. However, light-dependent effects were observed for A. solani, A. fumigatus and P. infestans, while B. cinerea and C. fulvum showed reduced sensitivity during the first two days. High-Cu coatings were most effective at inhibiting spore germination, whereas hyphal growth on multicomponent coatings may respond to different ions. Therefore, high-Cu, two-component coatings may be recommended for practical greenhouse applications. Full article

The fungal pathogen Botrytis cinerea (B. cinerea) attacks over 1400 plant species and results in estimated annual losses of $10–100 billion worldwide. In precision agriculture, deep learning (DL) provides reliable tools for rapid and objective plant disease detection. This study presents a unified two-stage DL solution for the automated detection of visible B. cinerea across three major vegetable crops—tomato, pepper, and cucumber—using standard RGB imagery. In the first stage, a YOLOv11-based instance segmentation model accurately localizes leaf regions, achieving a localization accuracy of 87.3% as measured by mAP₅₀. In the second stage, an ensemble of 13 MobileViT variant models analyzes the segmented leaf regions and performs per-crop classification into healthy and infected leaves. The proposed system achieves an overall detection accuracy of 84.05%, with per-class detection of infected leaves at 88.61% for pepper, 82.68% for tomato, and 70.55% for cucumber, measured using the F1-score. These results demonstrate that the proposed approach can reliably detect B. cinerea symptoms across different crops using only RGB data, offering a practical path toward smartphone-based field deployment and integration into decision support systems for timely, symptom-based disease management. Full article

Background/Objectives: The 3-ketoacyl-CoA synthase (KCS) enzyme is a key and rate-limiting component in the biosynthesis of very long-chain fatty acids (VLCFAs). Through controlling VLCFA production, KCS plays an essential role in plant cuticle formation. The necrotrophic fungus Botrytis cinerea can infect all aboveground parts of rose plants (flowers, leaves, and stems), causing severe economic losses. KCS restricts pathogen invasion by influencing cuticle formation and enhances tolerance to environmental stresses. While the KCS gene family has been well-studied in some plants, it remains unexplored in rose (Rosa × hybrida Hort.), a species of significant ornamental and economic value. Methods: In this study, we conducted a genome-wide analysis of the RcKCS gene family in rose, identifying 18 non-redundant genes. Phylogenetic, structural, and synteny analyses were performed to investigate the evolutionary relationships, gene architecture, and duplication events. The expression patterns of RcKCS genes in rose petals during B. cinerea infection were examined, and transient overexpression and silencing of RcKCS6 were used to study its function. Results: RcKCS6 was found to be upregulated during gray mold infection, and transient overexpression reduced lesion size on infected petals. Conclusions: Our study provides the first comprehensive analysis of the RcKCS gene family in rose and highlights RcKCS6 as a potential candidate for improving resistance to gray mold in rose through molecular breeding. Full article

To identify fungicide lead compounds with novel scaffolds and high efficacy, 33 novel bacillamide–acylhydrazone derivatives were successfully designed and synthesized by using a molecular hybridization strategy. The bioassay results showed that most of the target compounds exhibited promising inhibitory activity against B. cinerea. Among them, compound BAD-15 displayed the most potent antifungal activity with an EC₅₀ value of 6.725 μg/mL. Furthermore, preliminary SAR analysis revealed that the R group in hydrazine fragments exerts a significant influence on antifungal potency. Studying the molecular mechanism by morphological observation and transcriptome analyses revealed that BAD-15 may inhibit the activity of copper ion transmembrane transporters, leading to disrupted copper ion homeostasis and subsequent suppression of fungal growth. The present work indicates that bacillamide–acylhydrazone has potential as a novel scaffold for the development of fungicides, and compound BAD-15 may serve as a potential lead compound for the further development of novel fungicides. Full article

Monilinia laxa and Colletotrichum fioriniae are major fungal pathogens causing brown rot and anthracnose in stone fruits and shell fruits, leading to significant economic losses. Chemical fungicides are widely applied but can result in resistance development, environmental contamination, and food safety concerns. Biological control using entomopathogenic bacteria (EPB) of the genus Photorhabdus has emerged as an eco-friendly alternative. This study evaluated the in vitro antifungal activity of selected Photorhabdus species (P. kayaii 1723B, P. temperata 3017, P. cinerea 3086, P. laumondii 3196, and P. thracensis 3210) against M. laxa (M3) and C. fioriniae (VV081) using drop-to-drop confrontation and poisoned agar assays. Effects of fermentation time, preparation mode (original vs. centrifuged and filtered), and concentration (5, 10, 20%) were examined. Species-specific inhibition was observed, with Median Inhibition Index values indicated relatively higher antifungal activity for P. thracensis 3210 against M. laxa (0.718) and C. fioriniae (0.552), followed by P. cinerea 3086 (0.643 and 0.552) and P. kayaii 1723B (0.629 and 0.541). Fermentation time and preparation mode influenced antifungal activity in a strain-dependent manner, with longer fermentation periods and original culture preparations generally showing stronger inhibitory trends. Higher concentrations, especially 20%, were often associated with increased inhibition, although the magnitude of these effects varied among strain–pathogen combinations. Overall, these findings demonstrate that the strain- and pathogen-specific nature of antifungal responses in Photorhabdus, supporting their potential as components of targeted biological control strategies rather than uniform broad-spectrum agents. Full article

Phytopathogenic fungi pose a critical threat to global food security through substantial pre- and post-harvest crop losses, intensified by climate change and fungicide resistance. To address this, we synthesized low-concentration chitosan–inorganic oxide nanocomposites (CS-SiO₂, CS-ZnO, CS-CuO) via ionic gelation, a green and scalable method. Comprehensive characterization (DLS, UV-Vis, FTIR, XRD, SEM) confirmed nanocomposite formation, CS-SiO₂ exhibited uniform particle sizes (200–250 nm), while CS-CuO showed slightly larger particles, all with excellent dispersity. Zeta potential analysis confirmed strong colloidal stability, with pure chitosan nanoparticles (CSNPs) displaying a surface charge of +12.9 mV, while all nanocomposites retained positive charges, enhancing adhesion to negatively charged fungal membranes. In vitro antifungal assays against Alternaria alternata, Botrytis cinerea, Colletotrichum graminicola, and Fusarium graminearum demonstrated hierarchical efficacy: CS-CuO > CS-ZnO > CS-SiO₂, with CS-CuO achieving >80% growth inhibition against B. cinerea and A. alternata. SEM revealed severe hyphal damage and spore collapse in CS-CuO-treated fungi, attributed to synergistic reactive oxygen species (ROS) generation and chitosan-mediated membrane disruption. In vivo trials on B. cinerea-infected apples showed CS-CuO reduced lesion area by 81% and elevated host defense markers, including a 1.5-fold increase in total phenolic content and higher DPPH radical scavenging activity compared to controls. These nanocomposites, particularly CS-CuO, offer a sustainable, dual-action solution direct antifungal activity and enhanced host resilience while minimizing environmental impact. By integrating scalable synthesis, eco-compatibility, and efficacy, this work advances chitosan–inorganic oxide nanocomposites as viable alternatives to conventional fungicides, with immediate potential for agricultural and postharvest applications. Full article

During this phytochemical study, 13 compounds from the bark of Salix cinerea L. were isolated and their structures elucidated. These included two salicylic alcohol derivatives, one flavonol, two phenylpropanoids, two flavan-3-ols, two dimeric procyanidins, two dimeric prodelphinidins, and a unique ester of catechin (3-O-(1-hydroxy-6-oxo-2-cyclohexen-1-carboxylic acid), HCH-catechin). Furthermore, seasonal variations in the composition of Salix cortex regarding proanthocyanidins (PA) and the degree of polymerization were examined using NMR spectroscopy, revealing an increase in polymerization throughout the growing season 2020 associated with a consistent hydroxylation pattern in the B-ring. The isolated HCH-catechin was tested in vitro for its inhibitory effect on TNF-α-induced ICAM-1 expression in human microvascular endothelial cells (HMEC-1). A 24 h treatment with a 25 µM solution of HCH-catechin significantly reduced ICAM-1 expression (83.7 ± 3.2%) compared to unsubstituted catechin (97.9 ± 4.4%). Additionally, during a mass-spectrometric screening, numerous HCH adducts within the PA fraction could be identified, allowing for the proposition of a characteristic fragmentation pattern. This study establishes a foundation for a comprehensive assessment of the phenolic, PA-rich fraction in willow bark, particularly the occurrence of HCH adducts, which may contribute to the medicinal properties of Salicis cortex. Findings on seasonal variations and mass spectrometric profiling offer new insights into the quality standards for Salicis cortex as a medicinal remedy. Full article

Botrytis cinerea, the causal agent of grey mold in grapevine, remains one of the most economically important pathogens in viticulture and a key target for sustainable biocontrol strategies. This study evaluated the antagonistic potential of seven Trichoderma isolates (T1–T7), collected from the rhizosphere of grapevine in Morocco, using a combination of in vitro and in planta assays designed to capture multiple direct and indirect modes of action. The isolates exhibited variable levels of antagonism through competition, volatile organic compounds, extracellular metabolites, and elicitation responses. Preliminary in planta assays on detached grape berries further demonstrated that all selected isolates reduced lesion development, with preventive applications yielding the strongest protection. Overall, the study highlights the complementary and strain-specific mechanisms underlying Trichoderma & B. cinerea interactions and underscores the importance of isolate selection and application timing for the development of effective and environmentally friendly grey mold management strategies. These findings provide a mechanistic basis for the future evaluation of promising isolates under vineyard conditions. Full article

Botrytis cinerea poses a major threat to postharvest strawberries, causing significant losses due to gray mold. As a plant-derived antifungal agent, Rhodiola rosea L. essential oil (REO) possesses considerable healthcare benefits. However, its effectiveness and underlying mechanisms in the maintenance of postharvest products remain poorly understood. This study demonstrated that REO at 0.5 µL/mL completely inhibited the growth of B. cinerea under in vitro conditions. In vivo fumigation treatment with REO alleviated the severity of gray mold in strawberry fruit. Additionally, REO decreased natural decay and positively impacted marketability, as evidenced by higher firmness, total soluble solids, and ascorbic acid contents, as well as more favorable color attributes. Further investigations involving scanning electron microscopy, calcofluor white (CFW) staining, propidium iodide (PI) staining, 2′,7′-dichlorodihydrofluorescein diacetate assay, and cellular leakage tests were conducted to investigate the effects of REO treatment on gray mold mycelium. Results showed that REO treatment induced severe morphological distortions and collapse of mycelium. Within 30 min of exposure, REO triggered a sharp increase in PI fluorescence accompanied by a decrease in CFW fluorescence, without inducing an elevation in intracellular reactive oxygen species levels. The elevated leakage of nucleic acids and soluble proteins further confirmed that REO compromised the integrity of the cell barrier in B. cinerea. Collectively, these findings indicate that REO exerts potent antifungal activity by disrupting the integrity and functionality of B. cinerea cellular barriers, thereby reducing postharvest decay and positively impacting the marketability of strawberry fruit. Taken together, our findings suggest that REO represents a promising natural alternative for environmentally sustainable postharvest protection of strawberries. Full article

Botrytis cinerea poses severe postharvest losses in horticultural products, while synthetic fungicides raise food safety concerns. This study developed a GRAS-compliant antifungal strategy using vapor-phase Cymbopogon citratus essential oil (EO). GC-MS revealed citronellal (17.06%) as the dominant bioactive compound. The EO exhibited superior vapor-phase activity against B. cinerea, with EC₅₀ of 14.69 µg/mL (mycelial growth) and MIC of 7.81 µg/mL (spore germination), significantly lower than direct-contact efficacy (p < 0.05). Mechanistic analysis revealed a tripartite mode of action—rapid membrane disintegration (48% electrolyte leakage within 4 h), suppression of ROS defense enzymes (SOD/CAT/POD inhibition > 50%), and disruption of mitochondrial energetics (SDH activity reduced by 58.1%)—which induced irreversible cellular collapse. This multi-target strategy mitigates resistance development, a key limitation of single-mode fungicides. In commercial-scale trials, EO fumigation (125 µg/mL) reduced cherry tomato decay by 81.9–92.6% during 28-day storage, while maintaining firmness (15.9% higher than control) and nutritional quality (titratable acidity (TA) and total sugar content (TSC)). Notably, the vapor-phase EO also exhibited potent inhibitory activity against the spore germination of rubber tree powdery mildew (EC₅₀: 3.19 µg/mL), demonstrating its broad-spectrum antifungal potential. This finding significantly expands the application scope of C. citratus EO from postharvest preservation to preharvest crop protection. This work provides a scalable, residue-free alternative to synthetic fungicides for industrial postharvest applications. Full article

Endophytic fungi from the Cactaceae family are an underexplored source of bioactive secondary metabolites with potential applications in sustainable agriculture. This study investigated an endophytic fungus obtained from healthy cladodes of Opuntia ficus-indica growing in the Chilean Andean Precordillera. The influence of culture conditions, specifically pH and nitrogen concentration, on the production of diffusible and volatile antifungal compounds against the phytopathogen Botrytis cinerea was evaluated using dual-culture (confrontation) and sandwiched Petri dish assays. Morphological characteristics and molecular analyses confirmed that the isolate belongs to the genus Alternaria. Antifungal activity increased significantly under acidic conditions and limited nitrogen availability. The strongest inhibition by volatile compounds occurred at pH 4.5 and the lowest concentration of ammonium tartrate. Furthermore, ethyl acetate extracts at 40 mg/L obtained from Alternaria sp. cultures grown at pH 4.5 with 2.3 g/L ammonium tartrate inhibited B. cinerea mycelial growth by 60%. The study provides a framework for improving the yield of antifungal metabolites produced by Alternaria, contributing to the development of biofungicides for gray mold control. Full article

Biocontrol, a practice for using living organisms to target plant pathogens, offers a promising, sustainable agricultural strategy. This study involves epiphytic yeasts isolated from Vitis vinifera ssp. sylvestris and ssp. vinifera as natural antagonists against Aspergillus carbonarius, Botrytis cinerea, and Penicillium expansum. Twenty-one of 37 yeasts were chosen based on the Pathology Intensity (PA) score during preliminary in vivo screening. Following identification, dual-culture assays, VOC production, copper tolerance, and commercial fungicide resistance were assessed. On YPD and GJ medium, Saccharomyces isolates were the strongest antagonists, whereas P. terricola UMY197 inhibited Penicillium and Aspergillus. H. uvarum UMY1473 was notably effective against B. cinerea. VOC analysis confirmed that S. cerevisiae UMY1430 was the most effective against Aspergillus, likely owing to its production of oxalic acid, while S. cerevisiae UMY1438 was a producer of various esters and phenylethyl alcohol. C. intermedia UMY189, M. pulcherrima UMY1472, H. uvarum UMY1473, and S. cerevisiae UMY1436 were the most copper-resistant. Yeast activity on chemical fungicide SWITCH (up to 1 g/L) depended on culture media usage; in fact, a higher viability on YPD than on GJ was observed, where only 4 yeasts were able to grow. Thus, since several yeasts exhibit promising inhibitory activity through various mechanisms and against different molds, the use of synthetic consortia could represent a powerful and essential tool in field trials to limit fungicide use while preventing the emergence of resistance. Full article