Botrytis Cinerea Research Articles

Comprehensive Analysis of Volatile Organic Compounds and Their Impact on Apple Quality Following Some Essential Oil Treatments Against Botrytis cinerea

The susceptibility of apples to post-harvest decay by the fungus Botrytis cinerea has prompted innovative research into alternative preservation methods. In this regard, essential oils (EOs) have emerged as promising candidates due to their natural origin and potential antimicrobial properties. Investigating the biological significance of EO blends is crucial for understanding their potential antimicrobial mechanisms and evaluating their capacity to modulate metabolic responses that could inhibit post-harvest fungal decay in fruit tissues. This study delves into the intricate chemistry of apples when subjected to various EO treatments, shedding light on the profound changes in alcohols, esters, C6 compounds, terpenes, and volatile compounds. Based on our results, terpene concentrations exhibit significant variations with α-Pinene ranging from 13.4 µg L−1 in Fungus + Thymol + 1,8-Cineole treatment (Fun+Thy+Cin) to 28.7 µg L−1 in Fungus + Thymol + 1,8-Cineole + Eugenol treatment (Fun+Thy+Cin+Eug), and β-Pinene concentrations spanning 19.3 µg L−1 in Fungus + 1,8-Cineole + Eugenol treatment (Fun+Cin+Eug) to 45.5 µg L−1 in Fungus + Thymol + 1,8-Cineole + Eugenol treatment (Fun+Thy+Cin+Eug). Ester elaboration presents marked changes, with ethyl octanoate peaking at 715.7 µg L−1 in Fungus + Thymol + 1,8-Cineole + Eugenol treatment (Fun+Thy+Cin+Eug) and ethyl propionate reaching 152.9 µg L−1 in Fungus + Thymol treatment (Fun+Thy). The volatile compound dynamics also demonstrate significant variations, with hexanoic acid concentrations ranging from 0.1 to 0.2 among treatments and 3-Methylbutanal displaying concentrations from 0.8 to 6.4, with the highest concentration observed in the Control. The essential oil combination of Thymol, Eugenol, and 1,8-Cineol (Fun+Thy+Cin+Eug) had the most significant impact on the volatile compound content in the fruits. The findings from this study unveil the intricate responses of apple chemistry to various EO treatments. These insights hold promise for enhancing post-harvest apple preservation strategies through the modulation of EO treatments.

Capsicum annum NAC transcription factor CaNAC12 functions as a positive regulator of the stress response to salt, drought, and Botrytis cinerea in Nicotiana benthamiana

Capsicum annum NAC transcription factor CaNAC12 functions as a positive regulator of the stress response to salt, drought, and Botrytis cinerea in Nicotiana benthamiana

A LysM Effector Mediates Adhesion and Plant Immunity Suppression in the Necrotrophic Fungus Botrytis cinerea.

LysM effectors are suppressors of chitin-triggered plant immunity in biotrophic and hemibiotrophic fungi. In necrotrophic fungi, LysM effectors might induce a mechanism to suppress host immunity during the short asymptomatic phase they establish before these fungi activate plant defenses and induce host cell death leading to necrosis. Here, we characterize a secreted LysM protein from a major necrotrophic fungus, Botrytis cinerea, called BcLysM1. Transcriptional induction of BcLysM1 gene was observed in multicellular appressoria, called infection cushions, in unicellular appressoria and in the early phase of infection on bean leaves. We confirmed that BcLysM1 protein binds chitin in the fungus cell wall and protects hyphae against degradation by external chitinases. This effector is also able to suppress the chitin-induced ROS burst in Arabidopsis thaliana, suggesting sequestration of chitooligosaccharides in apoplast during infection. Moreover, contribution of BcLysM1 in infection initiation and in adhesion to bean leaf surfaces were demonstrated. Our data show for the first time that a LysM effector can play a dual role in mycelial adhesion and suppression of chitin-triggered host immunity, both of which occur during the early asymptomatic phase of infection by necrotrophic fungi.

Caffeic acid improves biocontrol effect of Pantoea jilinensis D25 against tomato gray mold.

Gray mold in tomato caused by Botrytis cinerea is a destructive disease, which can be treated using biocontrol agents. Pretreatment of biocontrol strains with oxidative-stress-ameliorating compounds can enhance their tolerance to oxidative microenvironment in infected fruit wounds. In this study, we aimed to determine the effect of caffeic acid (CA) on the biocontrol efficacy of Pantoea jilinensis D25 in gray mold in cherry tomato. D25 strain pretreated with CA exhibited enhanced oxidative stress response in cherry tomato wounds. CA-treated D25 cells recovered from cherry tomato wounds had lower ROS level and cellular oxidative damage than D25 cells not treated with CA. The antioxidation-related genes (CAT and GPX) were upregulated and colonization ability of D25 strain was improved in cherry tomato wounds after CA application. Furthermore, the biocontrol effects of CA-treated D25 strain were improved. CA pretreatment to D25 strain could significantly improve the expression of defense-related genes (SlMYC-2, SlLoxD, SlEIN2, and SlEIN3) and activity of defense-related enzymes in cherry tomato wounds. Application of CA to D25 strain could improve its ability to combat antioxidant stress and biocontrol efficacy against postharvest tomato gray mold. © 2024 Society of Chemical Industry.

Composition and antimicrobial activity of hydroalcoholic extracts of Pleurotus eryngii var. ferulae and P. eryngii var. elaeoselini.

The basidiomycetes Pleurotus eryngii var. ferulae Lanzi and P. eryngii var. elaeoselini Venturella et al. belong to the P. eryngii species complex, acting as facultative biotrophs in association with members of Apiaceae family, i.e., Ferula communis L. and Elaeoselinum asclepium L., respectively. The consumption of these fungi has rapidly increased in recent decades, not only thanks to their nutritional properties and pleasant flavor, but also for their bioactive and medicinal properties. A quantitative study of their hydroalcoholic extracts was carried out by liquid chromatography-mass spectrometry. The potential antimicrobial activity of the extracts was also tested against some phytopathogenic bacteria [Clavibacter michiganensis and Bacillus megaterium (Gram-positive), Pseudomonas viridiflava, Xanthomonas campestris, and Escherichia coli (Gram-negative)] and fungi (Aspergillus fumigatus, Penicillium italicum, Monilinia laxa, Botrytis cinerea, Cadophora sp., and Sclerotinia sclerotiorum). The chemical analysis allowed the identification of secondary metabolites belonging to different classes, as flavonoids, organic acids, amino acids, carbohydrates, vitamins, nucleic acids, fatty acids, and triterpenoids. Both extracts demonstrated antimicrobial activity against of the most tested microorganisms. The results can broaden the knowledge on the possible use of these fungal species in the agricultural sector.

Synthesis and Fungicidal Activities of Coumarin Derivatives as Succinate Dehydrogenase Inhibitors.

Succinate dehydrogenase inhibitors (SDHIs) have been developed to the fastest growing family of fungicides. To develop novel SDH inhibitors, twenty-seven novel non-amides coumarin derivatives were designed, synthesized and characterized .The bioassay revealed that most of the target compounds exhibited significant antifungal activity against Botrytis cinerea in vitro. Notably, compounds 1a and 2c with EC50 values of 0.92 μg/mL and 0.52 μg/mL, respectively，which were better than that of positive control chlorothalonil (EC50 =3.14 μg/mL).Moreover, in vivo antifungal assay results showed that compound 2c could observably inhibit the growth of B. cinerea on Kuerla pears with remarkable protective at a dosage of 200 μg/mL. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) investigation indicated that compound 2c significantly damaged the cell structures of B. cinerea mycelium. 3D-QSAR models were analyzed structure-activity relationships of all target compounds. Furthermore, molecular docking revealed that compound 2c was able to bind closely to the receptor protein of SDH. These results demonstrated that compound 2c may be potential candidate for novel SDH inhibitors.

Exogenous dsRNAs against chitin synthase and glucan synthase genes suppress the virulence of the pathogenic fungus Botrytis cinerea

Exogenous dsRNAs against chitin synthase and glucan synthase genes suppress the virulence of the pathogenic fungus Botrytis cinerea

2-Methylbutyric acid functions as a potential antifungal fumigant by inhibiting Botrytis cinerea and inducing resistance to gray mold in cherry tomatoes

Botrytis cinerea causes gray mold disease on postharvest fruit, resulting in serious economic losses. Fumigation with natural volatile organic compounds can replace artificially synthesized fungicides to control postharvest fungal decay of agricultural products. The antifungal effects and potential mechanisms of microbial volatile compound 2-methylbutyric acid (2-MBA) against B. cinerea were investigated in this study. Results showed that 2-MBA inhibited mycelial growth and spores germination, altered mycelial morphology and reduced metabolic vitality of B. cinerea. Moreover, 2-MBA induced oxidative damage by stimulating ROS accumulation, increasing malondialdehyde levels, and decreasing the ratio of reduced/oxidized glutathione. Further studies indicated that 2-MBA reduced mitochondrial membrane potential, interfered TCA cycle by decreasing succinate dehydrogenase activity and ATP content, ultimately leading to decreased mitochondrial activity. In addition, key genes involved in botrydial biosynthesis and mycelial growth were down-regulated. 2-MBA controlled postharvest gray mold by inducing resistance to B. cinerea in cherry tomatoes. Taken together, 2-MBA is an effective and promising fumigant to control gray mold caused by B. cinerea.

A Single Tube RPA/Cas12a Based Diagnostic Assay for Early, Rapid and Efficient Detection of Botrytis cinerea in Sweet Cherry.

The fungus Botrytis cinerea is a devastating plant pathogen that causes gray mold rot in many fruits and vegetables, leading to substantial losses globally. The pathogen induces a broad range of symptoms, making it challenging to differentiate from other pathogens, especially when plants exhibit no symptoms during visual inspection. Therefore, an efficient early warning and rapid detection system is crucial for surveillance and monitoring plant health, as well as for quantitative pathogen risk assessment to reduce microbial risks. In this study, we developed a single-tube detection method for B. cinerea based on RPA/Cas12a (Bc-RPA/Cas12a), which can be performed under field conditions and visualized using mini-UV light and ordinary mobile phone light within 45 minutes at a constant temperature. The Bc-RPA/Cas12a assay can detect B. cinerea without cross-reactivity with other pathogens and without compromising sensitivity. The universality of the Bc-RPA/Cas12a assay was demonstrated through its adaptability across different geographical regions of China. Furthermore, field simulation assays revealed that fruits inoculated with the target pathogen showed fluorescence, while fruits inoculated with non-targeted pathogens and double-distilled water (ddH2O) did not exhibit fluorescence. The findings suggest that this assay can be applied directly in field conditions for rapid and accurate detection of B. cinerea without the need for specialized equipment.

Lipopeptides from Bacillus atrophaeus MCM61 and melatonin suppress gray mold-induced oxidative stress in cut roses (Rosa hybrida L.) and improve vase life.

Application of melatonin and lipopeptides (LPs) derived from Bacillus strains is considered an efficient strategy to control plant diseases atboth pre and postharvest stages. However, the combined application of melatonin and LPs has not been studied yet. Therefore, the present study presents the synergistic effect of melatonin and LPs produced by Bacillus atrophaeus strain MCM61 against gray mold disease and its impact on quality parameters and vase life of cut roses. The stems of cut roses alongwith flowers were dipped in the melatonin solution at concentration (0, 0.2, 0.4, 0.6, and 0.8mM) and the results indicated thatthe vase life of flowers treated with the melatonin was enhanced compared to control. Melatonin at 0.6mM concentration showed the highest vase life of cut roses at day 8. Synergistic treatment with melatonin and LPs of B. atrophaeus MCM61 revealed that MDA and H2O2 content showed the highest decrease in cut roses. Similarly, relative water content, total phenol content, GSH content, and defense enzymes i.e., APX, SOD, PPO, POD, and CAT activities were increased in cut roses treated with co-application of B. atrophaeus MCM61 LPsand melatonin compared to single treatments and control treatment. Furthermore, the longevity of cut roses was also improved in flowers treated with a combined application of MCM61 and melatonin compared to other treatments.

Chickpea NCR13 disulfide cross-linking variants exhibit profound differences in antifungal activity and modes of action.

Small cysteine-rich antifungal peptides with multi-site modes of action (MoA) have potential for development as biofungicides. In particular, legumes of the inverted-repeat lacking clade express a large family of nodule-specific cysteine-rich (NCR) peptides that orchestrate differentiation of nitrogen-fixing bacteria into bacteroids. These NCRs can form 2 or 3 intramolecular disulfide bonds and a subset of these peptides with high cationicity exhibits antifungal activity. However, the importance of intramolecular disulfide pairing and MoA against fungal pathogens for most of these plant peptides remains to be elucidated. Our study focused on a highly cationic chickpea NCR13, which has a net charge of +8 and contains six cysteines capable of forming three disulfide bonds. NCR13 expression in Pichia pastoris resulted in formation of two peptide folding variants, NCR13_PFV1 and NCR13_PFV2, that differed in the pairing of two out of three disulfide bonds despite having an identical amino acid sequence. The NMR structure of each PFV revealed a unique three-dimensional fold with the PFV1 structure being more compact but less dynamic. Surprisingly, PFV1 and PFV2 differed profoundly in the potency of antifungal activity against several fungal plant pathogens and their multi-faceted MoA. PFV1 showed significantly faster fungal cell-permeabilizing and cell entry capabilities as well as greater stability once inside the fungal cells. Additionally, PFV1 was more effective in binding fungal ribosomal RNA and inhibiting protein translation in vitro. Furthermore, when sprayed on pepper and tomato plants, PFV1 was more effective in reducing disease symptoms caused by Botrytis cinerea, causal agent of gray mold disease in fruits, vegetables and flowers. In conclusion, our work highlights the significant impact of disulfide pairing on the antifungal activity and MoA of NCR13 and provides a structural framework for design of novel, potent antifungal peptides for agricultural use.

Structural characterization, in-silico studies, and antifungal activity of 5-methylmellein isolated from endophytic Alternaria burnsii.

The present investigation focused on exploring the potential of fungal endophytes as a valuable source of bioactive compounds with diverse applications. The phenolic compound 5-methylmellein was isolated for the first time from Alternaria burnsii, an endophytic fungus associated with Morus alba Linn. The compounds were structurally characterized using comprehensive spectral analysis, including 1H-, 13C, and 2D-NMR, as well as HRESI-MS. The study investigated the antifungal activity of 5-methylmellein against several plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, Cercospora beticola, and Rhizoctonia solani. In vitro assays showed significant inhibition of various plant pathogenic fungi, and the IC50 values ranging from 34.59 ± 1.03a µg/mL to 44.76 ± 1.03b µg/mL against the tested fungi. In vivo experiments on apples and grapes revealed that 5-Methylmellein significantly reduced fruit decay caused by Botrytis cinerea. The wound incidence in the control group reached 95.78%, while the treated groups exhibited a reduction of 37.54% after 15days. These findings underscore the potential of 5-methylmellein as a potent antifungal agent, suggesting its eco-friendly application in agriculture for managing fruit decay.

UV-exposure decreases antimicrobial activities of a grapevine cane extract against Plasmopara viticola and Botrytis cinerea as a consequence of stilbene modifications-a kinetic study.

Stilbenoid extracts, such as those originating from grapevine by-products (e.g. canes), are of interest for use as biopesticides in vineyard owing to their antimicrobial activities. However, stilbenoids are unstable in the environment, especially under light. This study aimed to chemically characterize the effect of UV light on stilbenoids present in a grapevine cane extract (CE), and to evaluate the antimicrobial activities against two major grapevine pathogens (Plasmopara viticola and Botrytis cinerea) of grapevine extracts exposed to UV. Treatment with UV (365 nm) on a grapevine CE led to degradation of stilbenoids (up to 71% after 1 h). The stilbenoid stability depended on their chemical structure: only those possessing CC, as trans-resveratrol and trans-ε-viniferin, were affected with first their isomerization and secondly their oxidation/cyclization. As a consequence, UV-exposed extracts (UV-CEs) showed reduced antimicrobial activities against the two pathogens (mycelium and spores). For instance, regarding P. viticola, an UV-CE exposed during 4 h showed an almost total loss of its activity on oomycete development and a 2.4-fold inhibition of zoospore mobility in comparison to CE. For B. cinerea, the inhibition capacity of the same UV-CE was reduced by only 1.1-fold on mycelial development and by 3.2-fold on conidial germination compared to CE. UV light triggered modifications on the structure of bioactive stilbenoids, resulting in losses of their antimicrobial activities. Photoprotection of stilbenoids has to be considered in the perspective of using them in vineyards as biopesticides. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Yarrowia lipolytica MBC25: A promising yeast strain for biocontrol of Botrytis cinerea in preharvest and postharvest decay of strawberries

Gray mold disease, induced by Botrytis cinerea, is the main cause of strawberry decay. In the current study, the ability and mechanism of aquatic yeast, Yarrowia lipolytica MBC25, to control B. cinerea in strawberries were studied in vitro, in vivo, in the greenhouse, and during storage. In vitro and in vivo experiments revealed that 108 CFU/ml Y. lipolytica had the strongest inhibitory effect against B. cinerea. In vitro studies showed Y. lipolytica inhibited 22 and 85% of B. cinerea growth in dual culture and volatile metabolite tests, respectively. Y. lipolytica showed the capability to produce fungal pathogen cell wall degrading enzymes (protease, lipase, and chitinase), siderophores, and plant growth promoters (indole-3-acetic acid and gibberellin). Y. lipolytica could establish, survive, and proliferate on strawberry plants and fruit in the greenhouse and during storage. Inoculated yeast reduced gray mold incidence in the greenhouse by 80%, which was comparable to the commercial fungicide, while Y. lipolytica inhibited gray mold during postharvest fruit storage, which was more than commercial fungicide. Y. lipolytica inoculated fruit also showed higher contents of total phenol, flavonoids, and enzymes such as superoxide dismutase, peroxidase, and phenylalanine ammonia-lyase. These findings suggest that Y. lipolytica may act via volatile metabolites and induce defense systems in the host plant via higher bioactive compounds and antioxidant enzymes. Pre-harvest application of Y. lipolytica has been shown to be a promising alternative to chemical fungicides in the management of strawberry gray mold disease.

Phenolic acid compounds isolated from Zanthoxylum armatum inhibit gray mold by disrupting cellular structures, affecting energy metabolism, and inducing oxidative stress of Botrytis cinerea

Botrytis cinerea is a phytopathogenic fungus that utilizes multiple infection pathways and is known for causing gray mold on crops. This fungal infection results in the deterioration of crop quality and substantial economic losses. ZH-5, a phenolic acid compound derived from the methanol extract of the twigs of Zanthoxylum armatum DC., exhibited significant inhibitory effects on the mycelial growth, spore germination, and germ tube elongation of B. cinerea in in vivo experiments. The EC50 of ZH-5 was determined to be 58.96 mg L−1, which surpassed the effectiveness of commonly used carbendazim and was comparable to the plant-derived pesticide eugenol. Our study focused on investigating the mechanisms underlying the antifungal activity of ZH-5. Our findings have demonstrated that ZH-5 has the ability to disrupt the integrity of cellular structures, including mitochondria. This disruption affects energy metabolism and induces oxidative stress, ultimately ZH-5 might be interfere with cellular activity or inhibit growth. In summary, we reported that ZH-5 demonstrated efficacy as a plant-derived compound against B. cinerea, making it a potentially valuable environmentally-friendly chemical agent for the management of gray mold.

Optimizing encapsulation of garlic and cinnamon essential oils in silver nanoparticles for enhanced antifungal activity against Botrytis cinerea pathogenic disease

The application of nanotechnology offers an environmentally sustainable approach to managing plant diseases and improving the protection of agricultural products and food. The antimicrobial potential of essential oils (EOs) derived from Allium sativum (garlic) and Cinnamomum verum (cinnamon) can be significantly enhanced through the application of nanotechnology. In this study, we encapsulated the EOs of A. sativum and C. verum in silver nanoparticles (AgNPs), which resulted in increased fungicidal activity against Botrytis cinerea, the pathogen responsible for anthracnose in many horticultural crops. The encapsulated EOs demonstrated a synergistic effect, achieving over 80% inhibition of B. cinerea mycelium growth and suppressing conidia germination by 75%. Furthermore, substantial morphological changes in the fungal hyphae were observed upon application of the nano-encapsulated EOs. The observed morphological changes included disrupted cell walls and deformed hyphal structures, which indicated severe damage to the fungal cells. The AgNPs function as carriers, enhancing the stability and dispersion of EOs and facilitating a more uniform and sustained release of the active compounds. The results of this study indicate that nano-encapsulated EOs have the potential to be effective antifungal agents, offering a promising approach for pathogen control in agriculture. The integration of nanotechnology with traditional antimicrobial agents enables the development of advanced formulations that provide enhanced protection against a wide range of plant pathogens. This approach not only enhances the efficacy of existing treatments but also reduces the necessity for synthetic chemicals, thereby aligning with sustainable agricultural practices.

Characterization of Tomato Seed Endophytic Bacteria as Growth Promoters and Potential Biocontrol Agents.

Endophytic bacteria residing within plant seeds are increasingly recognized for their potential to enhance plant growth and provide biocontrol against pathogens. Despite this, seed-borne endophytes remain underexplored in many crops, including tomato. In this study, we isolated and characterized bacterial endophytes from tomato seeds and evaluated their plant growth-promoting traits and antifungal activities. The taxonomic analysis of the Hawaii 7996 tomato seed endophyte collection revealed a diverse community, predominantly from the phylum Bacillota, with Paenibacillaceae and Bacillaceae as the most abundant families. Among the 35 unique strains identified, 19 produced indole-3-acetic acid, four exhibited siderophore production, and 12 could solubilize phosphate. These traits contribute to growth promotion and disease suppression in plants. In the plant growth promotion assay, several bacterial strains, notably Streptomyces olivaceus (BHM1), Streptomyces variegatus (BHM3), Bacillus stercoris (BHR2), and Moraxella osloensis (YHT4-1), demonstrated significant potential for tomato cultivation by positively affecting fresh weight, stem length, and root length. These strains consistently promoted growth across all three parameters evaluated in this study. Furthermore, several strains exhibited strong antifungal activity against major tomato pathogens, including Fusarium oxysporum race 1 and 2, and Botrytis cinerea. Notably, Bacillus subtilis (BHN1), Bacillus stercoris (BHR2), and Paenibacillus peoriae (YHR2-1) showed broad-spectrum antifungal efficacy. Our findings highlight the potential of seed-associated endophytic bacteria as growth promoters and biological control agents, offering promising avenues for sustainable agricultural practices.

Effectiveness of Nano-emulsions and Essential Oil of Fennel and their Major Components against Botrytis cinerea

Effectiveness of Nano-emulsions and Essential Oil of Fennel and their Major Components against Botrytis cinerea

Design, Synthesis and Fungicidal Activity of N-(5-Phenyl-1,3,4-Thiadiazol-2-yl)-N-((1-Phenyl-1H-1,2,3-Triazol-4-yl) Methyl) Benzamide Derivatives

Objective: Plant pathogens are serious hazards to agricultural production, but existing drugs have problems such as increased resistance of pathogenic bacteria and environmental pollution, and there is a need to find high-efficiency, low-toxicity fungicides with novel chemical structures. Methods: 5-phenyl-1,3,4-thiadiazol-2-amine was used as the starting material, and 20 new thiadiazole-linked compounds were designed and synthesized. Their in vitro inhibitory activities against plant pathogenic fungi were tested using a mycelium growth inhibition assay, and the relationship between the antifungal activity of the target compounds and their molecular structures was preliminarily discussed. Results: Bioassay results revealed that most of the title compounds exhibited excellent in vitro fungicidal activity, in which compound D16 showed the highest activity against Rhizoctonia solani (EC50 = 0.0028 μmol·L−1), D12 against Botrytis cinerea (EC50 = 0.0024 μmol·L−1), and D5 against Stemphylium lycopersici and Curvularia lunata (EC50 = 0.0105 μmol·L−1, 0.005 μmol·L−1). The structure activity relationship (SAR) was also discussed and the results are of great significance for the structural optimization and development of more effective thiadiazole antibacterial compounds. Conclusion: The results of this study are important for structural optimization and development of more potent thiadiazole antimicrobial compounds.

Novel pesticides design: co-encapsulation of citral and cyproconazole for the control of Botrytis cinerea using biocompatible nano-carriers.

Growing concerns about sustainability have driven the search for eco-friendly pest management solutions. Combining natural and synthetic compounds within controlled release systems is a promising strategy. This study investigated the co-encapsulation of the natural compound citral (Cit) and the synthetic antifungal cyproconazole (CPZ) using two water-based nanocarriers: solid lipid nanoparticles (SLNs) and chitosan nanoparticles (CSNPs). Both CSNPs and SLNs loaded with Cit + CPZ displayed superior antifungal activity against Botrytis cinerea compared to free compounds. Notably, CSNPs with a 2:1 Cit:CPZ ratio exhibited the highest efficacy, achieving a minimum inhibitory concentration (MIC100) of < 1.56 μg mL-1, lower than the 12.5 μg mL-1 of non-encapsulated compounds. This formulation significantly reduced the required synthetic CPZ while maintaining efficacy, highlighting its potential for environmentally friendly pest control. The successful co-encapsulation of Cit + CPZ within CSNPs, particularly at a 2:1 ratio, demonstrates a promising approach for developing effective and sustainable antifungal formulations against B. cinerea. © 2024 Society of Chemical Industry.