Search Results (897)

Bio-based coatings enriched with essential oils (EOs) represent a promising alternative to synthetic preservatives to extend strawberries’ shelf-life. This study evaluated the effects of chitosan (CHT) formulations containing three selected EOs (Illicium verum, Citrus sinensis, and Citrus limon) on the volatile profile, sensory quality, and antifungal activity of strawberry fruits. Volatile emissions were characterized by Headspace Solid Phase Micro-Extraction/Gas Chromatography-Mass Spectrometry, while sensory properties were assessed using Quantitative Descriptive Analysis. Antifungal activity was evaluated both in vitro and in vivo against Botrytis cinerea. Chitosan alone slightly modified the volatile profile, while EO-enriched coatings induced marked and concentration-dependent changes, reflecting the chemical composition of the incorporated EOs. Among the tested formulations, CHT combined with 1% C. sinensis EO provided the best balance between preservation of the characteristic strawberry aroma and overall sensory acceptance. In vitro assays showed that EO volatiles, particularly from C. sinensis and I. verum, significantly inhibited fungal growth, while diffusible compounds were less effective. In vivo, EO-containing coatings reduced disease incidence and severity by approximately 50%. These findings highlight the potential of CHT–EO coatings as sustainable options for postharvest preservation, although optimization of EO type and concentration is crucial to balance sensory quality and antimicrobial efficacy. Full article

Botrytis cinerea is a major phytopathogenic fungus responsible for substantial economic losses in horticultural crops, underscoring the need for sustainable alternatives to synthetic fungicides. This study investigated the influence of physical, chemical and biological culture parameters on the antifungal activity of culture filtrates produced by Trichoderma harzianum BOL-12QD. Culture conditions were sequentially optimised by evaluating light-filter exposure, carbon and nitrogen source composition, potato ecotype selection, co-cultivation with Botrytis cinerea, and volatile-mediated interactions. Antifungal activity was assessed using mycelial growth inhibition assays against Botrytis cinerea. Among the individual factors, violet-filter illumination, a medium containing 5 g L⁻¹ glucose and 250 g L⁻¹ potato extract, the Leke Pek’e potato ecotype, ammonium nitrate as nitrogen source, and co-cultivation with Botrytis cinerea at 10⁴ conidia mL⁻¹ produced the highest inhibitory effects. Sequential integration of these optimised conditions resulted in enhanced antifungal activity, reaching up to 62% inhibition. Volatile organic compounds produced by Trichoderma harzianum BOL-12QD exhibited only minimal antifungal activity under the conditions tested, suggesting that volatile-mediated antagonism plays a limited role in this system. In contrast, culture-dependent modulation of extracellular metabolite profiles was evidenced by comparative ¹H NMR fingerprinting, which revealed condition-specific spectral differences, with the optimised treatment displaying a distinct metabolic signature relative to all other conditions. Cytotoxicity assays in murine peritoneal macrophages showed no significant reduction in cell viability at concentrations up to 200 μg mL⁻¹. In vivo exposure to the optimised culture filtrate (250 mg kg⁻¹ d⁻¹ for 10 days) induced transient treatment-related clinical observations without mortality, indicating a need for further detailed toxicological characterisation. Overall, these findings demonstrate that the antifungal activity of Trichoderma harzianum BOL-12QD is strongly modulated by interacting environmental, nutritional and biological culture parameters. The results support the potential of optimised culture filtrates as a source of bioactive metabolites for biocontrol applications, while highlighting the importance of integrated biochemical and toxicological evaluation. Full article

Non-indigenous or alien macroalgae are increasingly recognized as ecological threats, sources of raw material, and reservoirs of bioactive compounds for industry and agriculture. This review analyses the valorization potential of this biomass, focusing on their antifungal and elicitor activities against phytopathogenic fungi, particularly Mediterranean (De Bary) Whetzel, 1945. The literature published since 2020 was retrieved from Scopus using targeted keyword combinations. Three major topics were examined: (i) invasive and beach-cast macroalgal and their ecological context, (ii) antifungal and elicitor properties of macroalgal extracts, and (iii) the use of deep eutectic solvents (DES) for the green extraction of bioactive compounds. Species such as Asparagopsis armata, Rugulopteryx okamurae, and Sargassum muticum have shown promising antifungal and elicitor effects, frequently associated with phenolic compounds and polysaccharides. Extracts from these algae can inhibit the growth of fungi or activate plant defense pathways, providing environmentally friendly alternatives to synthetic pesticides. Moreover, DES and natural DES (NADES) offer tunable, biodegradable solvents capable of efficiently extracting these bioactive molecules while reducing the environmental impact associated with conventional organic solvents. Overall, the valorization of this biomass represents a sustainable strategy that simultaneously mitigates ecological and economic impacts and contributes to the development of sustainable inputs in agriculture. Full article

Necrosis- and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) are conserved microbial proteins that contain immunogenic patterns capable of activating plant pattern-triggered immunity (PTI). NLP patterns from Sclerotinia sclerotiorum (SsNLPs), a destructive necrotrophic fungal pathogen with a broad host range, have been identified, and their roles in PTI have been revealed. Nevertheless, the molecular mechanisms by which SsNLPs stimulate plant immunity remain largely unknown. In this study, we phylogenetically characterized SsNLPs and demonstrated the involvement of the phytocytokine receptor-like kinases PEPRs in SsNLP1-triggered immunity. SsNLPs contained the NPP1 domain and GHRHDWE motif and were phylogenetically closely associated with Botrytis cinerea NLPs. SsNLP1 treatment strongly induced the expression of PEPR genes. Further genetic analyses using Arabidopsis wild-type, pepr1 pepr2 double mutant, and PEPR1 overexpression lines showed that SsNLP1 elicited diverse immune responses, including reactive oxygen species (ROS) accumulation and defense gene activation, and induced plant resistance to S. sclerotiorum. Notably, the induced plant resistance and immune responses were strengthened in PEPR1 overexpression lines and significantly reduced in the pepr1 pepr2 mutant, indicating a positive role of PEPR signaling in SsNLP1-triggered immunity. Overall, our results revealed that phytocytokine PEPR1 signaling amplifies PAMP SsNLP1-triggered immunity, thereby enhancing resistance against S. sclerotiorum. Our findings provide an example of the coordination between PAMP- and phytocytokine-triggered immunity for robust resistance to a necrotrophic pathogen. Full article

Diallyl trisulfide (DATS), a volatile natural sulfur-containing compound derived from garlic, possesses antifungal and preservative potential. However, its biocontrol efficacy against postharvest gray mold of tomato and the molecular mechanisms underlying fruit quality maintenance remain unclear. In this study, we systematically investigated the inhibitory effect of DATS fumigation on postharvest gray mold, its role in fruit quality maintenance, and the associated molecular mechanisms through in vitro antifungal assays, physiological and biochemical measurements, transcriptome sequencing, and correlation analysis. In vitro experiments showed that DATS at 50 μL L⁻¹ completely inhibited spore germination and germ tube elongation of Botrytis cinerea in a concentration-dependent manner, and disrupted spore membrane integrity (FDA-positive spores dropped from 73.4% to 2.9% at 50 μL L⁻¹). In vivo experiments demonstrated that Bc + DATS treatment completely inhibited lesion development compared to the control Bc, enhanced the activities of superoxide dismutase, catalase and peroxidase (e.g., CAT activity 2.20-fold higher than Bc on day 3), decreased malondialdehyde accumulation (0.65-fold of Bc on day 4), and delayed the declines in total soluble solids, titratable acidity, soluble sugars and vitamin C content (VC content 4.14-fold higher than Bc on day 4). Transcriptomic analysis revealed that DATS treatment up-regulated genes involved in plant hormone signal transduction, ubiquitin-mediated proteolysis, and phenylalanine metabolism, while down-regulating core MAPK kinases and histidine decarboxylase. Correlation analysis demonstrated significant associations between the expression of these pathway genes and antioxidant enzyme activities, vitamin C content, and lesion diameter. Collectively, DATS achieves effective control of postharvest gray mold and maintenance of fruit quality in tomato through direct antifungal activity, synergistic activation of hormone/MAPK signaling, reprogramming of phenylalanine metabolism, and modulation of membrane lipid homeostasis. This study provides a theoretical and practical basis for developing DATS as a green postharvest preservative to reduce food loss and ensure food safety. Full article

Phytopathogenic fungi cause major agricultural losses worldwide. Their control relies largely on synthetic fungicides, which raise concerns related to environmental impact, resistance development, and human health. Botanical extracts represent a promising, sustainable alternative, and members of the Brassicaceae family are recognized as rich sources of antifungal metabolites. In this study, the antifungal activity of ethanol extracts from Brassica napus and Capsella bursa-pastoris was evaluated against Botrytis cinerea, Colletotrichum acutatum, and Fusarium oxysporum, which are major phytopathogens widely recognized for causing significant diseases in diverse commercial crops worldwide. Antifungal effects on mycelial radial growth and conidial production were assessed in vitro using the amended culture medium assay. Extracts were chemically characterized by LC-DAD-ESI-MS, and a single-Y orthogonal partial least squares (OPLS) analysis was applied to integrate chemical and bioactivity data to identify metabolites associated with antifungal activity. Test botanical extracts showed organ- and pathogen-dependent antifungal activity. B. napus was more active, as seeds reduced B. cinerea growth, and roots strongly suppressed conidiation and inhibited F. oxysporum. In contrast, C. bursa-pastoris was less effective, with only the flower and fruit extracts causing modest reductions in F. oxysporum, while the other extracts were largely inactive. Fourteen metabolites were tentatively identified based on UV-Vis and mass spectral data. Among them, one flavanol and two indole-containing compounds were statistically correlated with antifungal activity, were subsequently isolated, and were structurally confirmed by NMR spectroscopy. These compounds (1–3) exhibited reasonable antifungal activity (IC₅₀ < 40 µM). The integrative covariate-based metabolic profiling approach proved operative for identifying bioactive constituents in the test botanical extracts, supporting the potential of Brassicaceae-derived extracts and their metabolites as natural antifungal agents. Full article

Peanut (Arachis hypogaea L.) production faces persistent threats from various infectious diseases. Planting healthy varieties with robust botanical defense networks is critical for minimizing future costs. Non-expressor of pathogenesis-related (NPR) regulators are involved in immune activation and act as key targets for deeper stress adaptation, and are thus promising targets for genetic enhancement. In this study, we characterized the peanut NPR4B protein and demonstrated its local subcellular binding to the nucleus. Ectopic overexpression of AhNPR4B in Arabidopsis thaliana significantly enhanced resistance to the necrotrophic pathogen Botrytis cinerea and enhanced cold tolerance, as supported by quantitative and statistical analyses (p < 0.05). As regards underlying molecular events, Y2H (Yeast 2-Hybrid) analysis revealed a binding in vitro physical relation of AhPR2-like to AhNPR4B. This binding was demonstrated in vivo through BiFC (Bimolecular Fluorescence Complementation). These results suggest that the AhNPR4B-AhPR2-like complex may act as a key regulatory module associated with biotic and abiotic stress signaling, potentially contributing to broad-spectrum stress resistance. These findings provide foundational insights into the functional roles of AhNPR4B and its interaction with AhPR2-like in regulating stress resistance and support its potential as a candidate target for future genetic improvements to enhance stress resilience in peanuts. Full article

Botrytis cinerea is a globally distributed phytopathogenic fungus that causes gray mold in eucalyptus seedlings, posing a severe threat to eucalyptus nursery production. Pseudomonas eucalypticola Liu et al. NP-1 is an endophytic bacterium isolated from eucalyptus with broad-spectrum antifungal activity. In this study, the fermentation broth extract of strain NP-1 was prepared using the organic solvent extraction method. The inhibitory effects, antifungal mechanisms, and biocontrol efficacy of the extract against B. cinerea were investigated. The results suggested that the NP-1 extract effectively inhibited mycelial growth, conidial germination, and germ tube development of B. cinerea. The EC₅₀ and EC₉₀ values for mycelial inhibition were 110 μg/mL and 332 μg/mL, respectively, while those for conidial germination inhibition were 126 μg/mL and 310 μg/mL. Microscopic and ultramicroscopic observations indicated that while the mycelial structures in the control and EC₅₀ groups remained intact, the EC₉₀ treatment significantly was associated with protoplasmic aggregation, leakage, and cavitation, suggesting potential structural damage to the fungal cells. In vitro and in vivo biocontrol assays showed that the control efficacy against gray mold reached 90.0% on detached eucalyptus leaves and 93.3% on eucalyptus seedlings. These findings elucidate the biocontrol potential of NP-1 and lay a foundation for the development of bio-based pesticides. Full article

Broccoli (Brassica oleracea var. italica) by-products represent an abundant and underutilized source of bioactive compounds with potential applications in sustainable food systems. This study aimed to characterize the microbiota associated with different plant fractions (leaves, stems, and heads) of broccoli (Parthenon and Tritón cultivars) and to evaluate the antioxidant and antimicrobial properties of their extracts, using cauliflower as a reference. Microbial counts and fungal identification (ITS sequencing) were performed, while phytochemical profiles were analyzed by HPLC-ESI-QTOF. Antioxidant activity was assessed using DPPH and ABTS assays, and antimicrobial activity under in vitro conditions was evaluated against selected foodborne bacteria and phytopathogenic fungi. Broccoli by-products, particularly leaves, showed lower microbial loads in certain cultivars and were rich in phenolic compounds and glucosinolates; however, higher phenolic content did not always correlate with greater antioxidant activity, highlighting the importance of compound composition. All extracts showed strong antibacterial activity at higher concentrations, especially against Listeria spp. Notably, antifungal activity was selective but relevant, with consistent inhibition observed against Alternaria alternata, while Penicillium purpurogenum and Botrytis cinerea exhibited higher resistance. Overall, these findings highlight the potential of broccoli by-products as sustainable sources of natural bioactive compounds for food applications, particularly in the development of preservation strategies and postharvest treatments. Further studies focusing on individual compounds and their specific biological activities are needed to better understand the mechanisms underlying these effects and to support their application in real food systems. Full article

Safflower (Carthamus tinctorius L.) is an economically important crop, yet its production is severely threatened by fungal diseases including Botrytis cinerea. The molecular mechanism underlying disease resistance in safflower remains largely unclear. Extracellular vesicles (EVs), as vital carriers for cross-kingdom communication and transport, play crucial roles in plant antifungal defense. Lipid transfer proteins (LTPs), members of the pathogenesis-related protein 14 family, have also been shown to be key players in plant disease resistance. The promising resistance-related candidate gene CtLTP8 was previously identified via genome-wide association study (GWAS). In this study, a genome-wide analysis of the LTP gene family in safflower was performed. EVs were isolated from the apoplastic washing fluid of B. cinerea-infected safflower leaves, and proteomic analysis was performed. Numerous proteins associated with disease resistance, including CtLTP8, were detected by proteomic profiling. CtLTP8 was found to be present in EVs through molecular biological experiments. Moreover, stable overexpression of CtLTP8 in safflower significantly increased resistance to B. cinerea. In summary, this study characterized the disease resistance-related proteome of safflower EVs, and verified the presence of CtLTP8 in EVs and its antifungal function, providing valuable gene resources and theoretical support for safflower disease-resistance breeding and research on EV-mediated plant immune mechanisms. Full article

Botrytis cinerea is a major phytopathogen responsible for significant postharvest losses in plant-derived foods. The increasing resistance to synthetic fungicides has driven the search for sustainable alternatives, including enzyme-based biofungicides. In this study, the proteolytic fraction P1G10 from Vasconcellea pubescens latex was encapsulated in an alginate–chitosan (ALG-CS) matrix to improve its stability and antifungal performance. The encapsulated formulation (ALG-CS-P1G10) retained ~95% enzymatic activity after 8 h under stress conditions (37 °C, 1350 lux), compared with 67% for the free enzyme. In vitro assays demonstrated a dose-dependent inhibition of B. cinerea growth, with an IC₅₀ value of ~11 mg/mL determined using a logistic model. At this concentration, the formulation reduced fungal adhesion by more than 80% and increased sensitivity to cell wall-disrupting agents (Congo Red and Calcofluor White), pointing to alterations in cell wall integrity. Importantly, the encapsulated system provided a more stable and sustained antifungal effect, consistent with a controlled-release mechanism. These results demonstrate that coupling enzyme stabilization with controlled release can improve the functional performance of protease-based antifungal systems, offering a promising strategy for the development of biofungicides in postharvest applications. Full article

Plant hormones and their synthetic analogueues offer sustainable alternatives for crop protection, yet the direct antifungal activity of strigolactone (SL) and its analogues against necrotrophic pathogens remain largely unexplored. Here, we screened eight phytohormones and related analogues for treatments of Botrytis cinerea and identified the SL analogue rac-GR24 (racemic GR24) as a concentration-dependent growth inhibitor active at low micromolar concentrations. Given the stereochemical complexity of SLs and their analogues, we evaluated multiple enantiopure isomers and found that ent-5DS and GR24^ent-5DS, which differ in configuration from natural SLs, exhibited the strongest inhibitory activity. This stereospecific response was further validated using another filamentous fungus, Sclerotinia sclerotiorum, which displayed an identical susceptibility profile. Combinatorial treatments with enantiopure isomers and double-concentration rac-GR24 revealed that the antifungal effect of the racemate is primarily attributable to the GR24^ent-5DS enantiomer, whereas the opposite enantiomer GR24^5DS is almost inactive. Collectively, our findings uncover a stereospecific response in fungal pathogens, demonstrating that B. cinerea and S. sclerotiorum respond to exogenous SL analogues in a chirally selective manner. This work establishes a stereochemically defined framework for developing enantioselective fungicidal agents with potential applications in sustainable agriculture. Full article

Cut roses (Rosa hybrida L.) are highly sensitive to postharvest conditions, often experiencing quality losses associated with declines in SPAD values (relative chlorophyll index), color instability, Botrytis cinerea incidence, and impaired bud opening. This study aimed to evaluate the effects of different storage atmospheres, including controlled atmosphere (CA; 10% CO₂ + 3% O₂ and 6% CO₂ + 3% O₂), normal atmosphere (NA), and modified atmosphere packaging (MAP; LDPE1 (low-permeability MAP): 25 µm, 8000 cc m⁻² day⁻¹ O₂ permeability; LDPE2 (high-permeability MAP): 25 µm, 12,000 cc m⁻² day⁻¹ O₂ permeability), on SPAD values, color parameters, disease incidence, and bud development in cut rose cultivars (Rosa hybrida L.) cvs. ‘Rhodos’ and ‘Athena’ harvested in May, June, August, and November. The experiment was conducted as a factorial completely randomized design with seven biological replicates per treatment, each consisting of a single flower. Treatments were applied in combination with 1-methylcyclopropene (1-MCP, 625 ppb) and a commercial postharvest hydrating solution (Chrysal RVB, 1 mL L⁻¹) under storage conditions of 0.5 ± 0.5 °C and 80–85% relative humidity. The results indicated that CA conditions in combination with 1-MCP maintained higher SPAD values, improved color stability, and were associated with lower Botrytis incidence (p < 0.01). In addition, the low-permeability LDPE1-based MAP treatment minimized variations in hue angle (h°) and improved bud development scores, while the hydrating solution treatment promoted bud opening, particularly in cv. ‘Athena’, although its effect on disease suppression was limited. Overall, the combined application of controlled atmosphere storage and 1-MCP generally showed superior performance in maintaining postharvest quality, reducing disease incidence, and preserving the visual and physiological attributes of cut roses, with effects varying depending on cultivar and evaluated parameter. Full article

An isolate of Peniophora lycii was obtained from grapevine, and its interactions with several grapevine-associated fungi and the plant host were examined. The fungus was not able to infect intact leaves, but necrotized the margins of foliar disks and caused necrosis and white rot in woody tissues. In dual cultures, P. lycii and Aureobasidium pullulans showed mutual antagonism. Mycoparsitism of P. lycii was observed on epiphytic Botrytis cinerea, Alternaria sp., and endophytic Botryosphaeria dothidea interaction partners. In contrast, P. lycii showed trophic growth towards endophytic Phaeomoniella chlamydospora without any signs of harmful interactions. Dual inoculations of foliar disks with epiphytic fungi revealed no effects of fungal interactions on necrosis development by pathogens and verified mycoparasitic interactions in planta. Co-infection assays of cuttings with endophytic pathogen fungi showed cumulative effects of fungal interactions on wood symptom expression, with the exclusive contribution of P. lycii to white rot development. In addition to providing the first isolation of P. lycii from grapevine and the description of its mycoparasitic behavior, the present study suggests that the fungus may act as an opportunistic grapevine pathogen, probably as a secondary colonizer in trunk diseases. The observed dual host preference may allow trunk disease pathogens to initially feed on fungi, followed by damage to the grapevine. This may be in connection with the exceptionally long latency of these syndromes. Full article

Papain-like cysteine proteases (PLCPs) are central immune hubs frequently targeted by pathogen effectors. Sugarcane smut, caused by Sporisorium scitamineum, threatens global sugarcane yield, yet effector manipulation of host PLCPs remains unclear. Genome-wide analysis of Saccharum spontaneum AP85-441 identified 61 PLCP-encoding genes, which were classified into nine conserved subfamilies. Among these, ScRD21A, a member of the RD21 subfamily, was prioritized for functional characterization. Two Pit2 homologs, SsPit2A and SsPit2B, were identified from S. scitamineum. Yeast two-hybrid, BiFC and pull-down assays demonstrated that both effectors interact with ScRD21A, and that this interaction depends on a conserved LXRR motif within their PID14-like region. In total protein extracts from Nicotiana benthamiana, co-expression of SsPit2A or SsPit2B reduced ScRD21A-associated cysteine protease activity. Transient expression of ScRD21A enhanced flg22-induced ROS production, attenuated Pst DC3000-induced hypersensitive response-associated necrosis, and increased resistance to Botrytis cinerea. Together, these results support a conserved PLCP-targeting strategy in smut fungi and identify the ScRD21A–SsPit2A/B module as a tractable framework for studying effector–protease interactions relevant to sugarcane smut. Full article