Search Results (52)

Botrytis cinerea Pers., the causal agent of grey mould, causes major economic losses in viticulture by reducing grape and wine quality and yield. Antagonistic yeasts that release bioactive volatile organic compounds (VOCs) represent a sustainable alternative to synthetic fungicides. Here, VOCs produced by Pichia guilliermondii strain ZIM624 were identified and assessed for antifungal activity against B. cinerea. 65 VOCs—including higher alcohols, volatile phenols, esters, and terpenes—were detected using two newly developed and validated analytical methods combining automated headspace solid-phase microextraction with gas chromatography–mass spectrometry. A total of 13 VOCs were selected for the bioassays. Fumigation assays demonstrated that terpenes (citronellol, geraniol, nerol, α-terpineol, and linalool) were the most effective inhibitors of B. cinerea mycelial growth (EC₅₀ = 6.3–33.9 μL/L). Strong inhibition was also observed for 4-vinylphenol and isoamyl acetate. In vivo assays confirmed that exposing infected grape berries to P. guilliermondii VOCs significantly reduced grey mould incidence. These results highlight the potential of P. guilliermondii ZIM624 volatiles as natural biofumigants for the eco-friendly management of B. cinerea in grapes. Future research should focus on optimising VOC production, evaluating efficacy under field conditions, and developing formulations for practical application in vineyards and post-harvest storage. Additionally, investigating potential synergistic effects of VOC combinations could lead to more effective biocontrol strategies. Full article

In response to the rising demand for sustainable engineering materials and waste valorisation strategies, this study investigates the multi-objective optimisation of eco-friendly hybrid composites reinforced with natural fibres and industrial waste. Sixteen composite specimens were fabricated using compression moulding by varying sisal fibre from 0 to 45 wt.%, banana fibre from 0 to 45 wt.%, NaOH alkali treatment from 0 to 6%, and red mud filler from 1 to 4 wt.%. Mechanical properties were evaluated following ASTM standards D256 for impact strength, D790 for flexural strength, D638 for tensile strength, D5379 for shear strength, and E18 for hardness. The Taguchi method combined with grey relational analysis was employed to identify optimal processing conditions. The best mechanical performance, with an impact strength of 6.57 J, flexural strength of 72.58 MPa, and tensile strength of 65.52 MPa, was achieved with 30 to 45 wt.% sisal fibre, 15 wt.% banana fibre, 6% NaOH, and 3 to 4 wt.% red mud. ANOVA revealed that NaOH treatment had the most significant influence on mechanical properties, with high F values and p values close to 0.05. Grey relational analysis proved more effective for multi-objective optimisation, with the highest grey grade of 0.894 observed in the specimen containing 45 wt.% sisal fibre, 6% NaOH, and 2 wt.% red mud. These findings highlight the critical role of fibre treatment and hybrid reinforcement in enhancing performance. The optimised composites demonstrate strong potential for use in automotive interior panel applications, offering a sustainable alternative with balanced strength and reduced environmental impact. Full article

Grey mould caused by Botrytis cinerea presents significant challenges to apple production including organic farming. Biocontrol yeasts and their consortia can limit fungal diseases. This study evaluates the efficacy of selected yeast isolates and their consortia in suppressing B. cinerea in stored apples. The yeast strains tested—Wickerhamomyces anomalus 114/73, Naganishia albidosimilis 117/10, and Sporobolomyces roseus 117/67—were assessed at 4 °C and 23 °C, individually and in consortia. The results demonstrate the superior efficacy of a consortium combining all three isolates, which achieved the highest reduction in spore germination and disease severity. A two-strain consortium of isolates 114/73 and 117/10 also showed substantial biocontrol activity, outperforming single-strain treatments. These combinations effectively suppressed B. cinerea growth and displayed rapid colonization of apple wounds. The study highlights the potential of yeast isolates and their consortia to manage postharvest fungal decay, addressing a critical need for sustainable, eco-friendly solutions in organic apple production. Full article

The grey mould fungus Botrytis cinerea is a dangerous plant pathogen responsible for substantial agricultural losses worldwide. The pathogenic mechanisms still have many unclear aspects, and numerous new pathogenic genes remain to be identified. Here, we show that the sterol regulatory element-binding protein Sre1 plays an important role in the development and pathogenicity of B. cinerea. We identified a homologue of gene SRE1 in the B. cinerea genome and utilized a reverse genetics approach to create the knockout mutant Δsre1. Our results demonstrate that SRE1 is essential for conidiation, as Δsre1 produced only 3% of the conidia compared to the wild-type strain. Conversely, Δsre1 exhibited increased sclerotium production, indicating a negative regulatory role of SRE1 in sclerotium formation. Furthermore, ergosterol biosynthesis was significantly reduced in the Δsre1 mutant, correlating with increased sensitivity to low-oxygen conditions. Pathogenicity assays revealed that Δsre1 had significantly reduced virulence, although it maintained normal infection cushion formation and penetration capabilities. Additionally, SRE1 was found to be crucial for hypoxia adaptation, as Δsre1 showed abnormal germination and reduced growth under low-oxygen conditions. These findings suggest that SRE1 mediates the development and pathogenicity of B. cinerea by regulating lipid homeostasis and facilitating adaptation to host tissue environments. Full article

The present study investigated the antifungal potential of hydroalcoholic extracts of propolis against the causal agent of grey mould, Botrytis cinerea, by in vitro and in vivo assays. Five different propolis from different Italian regions were subjected to hydroalcoholic extraction using different ethanol concentrations and extraction methods. The preliminary bio-assay showed significant inhibitory effects on B. cinerea mycelial growth of propolis extracts obtained using 90% ethanol and subjected to sonication. The calculation of EC50 values, based on the demonstrated efficacy of non-volatile and volatile metabolites of propolis extracts, was useful to understand the main fraction involved in the antifungal activity of the samples and to perform the in vivo assay on grape and blueberry fruits. Three of the propolis extracts showed a high amount of genistein. Conversely, the other two propolis showed a fair amount of apigenin, caffeic acid, chrysin, ferulic acid, kaempferol, luteolin, p-coumaric acid and quercetin. From the volatile analysis of propolis, the main compounds detected were α-cadinol, α-eudesmol, calamenene, cadinol, benzyl acetate, benzyl benzoate and benzyl alcohol. The results suggest that propolis extracts have potential as an effective postharvest antifungal treatment, with varying degrees of efficacy depending on the extraction method and the type of propolis metabolites. Full article

Botrytis cinerea, the grey mould fungus affecting over 1400 plant species, employs infection cushion (IC), a branched and claw-like structure formed by mycelia, as a critical strategy to breach host surface barriers. However, the molecular mechanisms underlying IC formation remain largely unexplored. In this study, we utilized a forward genetics approach to establish a large T-DNA tagged population of B. cinerea, which contained 14,000 transformants. Through phenotype screening, we identified 161 mutants with defects in IC development. Detailed analyses revealed that these mutants exhibited various degrees of impairment in IC formation, ranging from complete failure to form ICs to a reduction in the number and maturity of ICs. Further genetic analysis of one of the mutants led to the identification of EXO70, a gene encoding a component of the exocyst complex, as a key regulatory factor in IC development. Mutants with deletion of EXO70 failed to form ICs, confirming its crucial role in the process. The mutant library reported here provides a rich resource for further large-scale identification of genes involved in IC development. Our findings provide valuable insights into the genetic and molecular basis of IC formation and offer new targets for controlling B. cinerea pathogenicity. Full article

This article presents the results of experimental studies and numerical calculations that were conducted to analyse the phenomena that occur during the operation of an ingot mould that is designed for casting steel ingots. The studies were conducted on an experimental stand in a foundry on an ingot mould that was designed to make ingots that weigh up to six tons; they consisted of determining the temperature of the ingot mould and measuring the displacements of its walls during filling with steel and cooling. These studies were used to create and verify a numerical model that was used to determine the temperatures, displacements, deformations, and stresses in ingot mould walls during the operating cycle using the FEM method. Microstructure studies of ingot cast iron that was subjected to thermal fatigue were also conducted on a laboratory stand; the temperature changes and test times were the same as those used under the normal operating conditions of the ingot mould. Cast iron samples were subjected to heating and cooling cycles within a range of 0 to 60 cycles; then, tensile tests were performed to determine their stress–strain curves. As a result of the conducted tests, a great influence was found of the number of cycles on decreases in the values of the modulus of elasticity and tensile strength—especially within a range of 0 to 10 cycles. A relationship was also found between the changes in these values and the image of the cast iron microstructure. Based on images of the cast iron microstructure after being subjected to different numbers of thermal fatigue cycles, the mechanism of the crack initiation and propagation was determined. The influence of the changes in the strength of the cast iron and the stress state that was determined by the FEM method on the durability of the tested type of ingot mould was analysed. The obtained research results will be useful for introducing design changes that are aimed at increasing the fatigue durability of ingot moulds. Full article

Grey mould disease, caused by various Botrytis species, poses a significant threat to important plants worldwide. This study aimed to characterize Botrytis populations on strawberry and roses, economically relevant host plants, and raspberry, used as a representative of wild plants, in Iran. A total of 389 isolates were collected and analyzed based on morphological features and haplotyping using molecular markers, transposable elements (Boty and Flipper), and fungicide response. Moreover, 60 isolates were used for phylogenetic analysis based on the rpb2 gene, and 16 selected isolates from each clade were further characterized using the g3pdh, hsp60, and nep2 genes. The results revealed the presence of three distinct species, Botrytis cinerea, Botrytis sinoviticola, and Botrytis prunorum, among the sampled isolates. Additionally, this study reports for the first time the presence of B. sinoviticola on strawberry and isolates belonging to B. cinerea group S in Iran. These findings provide insights into the diversity and composition of Botrytis populations on Iranian host plants. Full article

This article deals with the possibility of using 3D-printed models as an input for the production of a mould for cast iron castings. This new progressive process is significantly faster (with sufficient accuracy) compared to the current way of making models for moulds. The need to create a wooden model is removed by this process. The quality of this wooden model was highly dependent on the experience and qualifications of the worker. This article describes the manufacturing process of the model and mould in detail. The key dimensions of the final parts are compared with the model and, thus, the accuracy of the chosen procedure is verified. A 3D-printing technology known as Multi Jet Fusion (MJF) was used to produce the model. The material used for the production of the model is polyamide PA12 with 40% glass fibre filling. This material has sufficient structural and strength properties to be used for the given application. Taking into account the dimensions of the part and the printing space of the printer, it was necessary to structurally modify and divide the part. The inlet cone of a turbine is used as an example This cone is produced from grey cast iron as standard. Full article

The evaluation of the potentiality of lactic acid bacteria (LAB) strains isolated from fermented products to inhibit Botrytis cinerea and Escherichia coli O157:H7 growth on spinach and lettuce was conducted. From a total of forty LAB strains tested, three were selected due to their high inhibitory effect on plant pathogenic fungi. The identification of these isolates based on a 16S rRNA gene fragment sequence analysis confirmed the genus of Levilactobacillus sp. and Lactiplantibacillus sp. An effective method of coating LAB isolates on the lettuce and spinach surface was developed. The leaves were immersed in bacterial suspension (5.0 × 10⁶ cfu mL⁻¹) for 4 s and drained on tissue paper. LAB survived on lettuce and spinach leaves for 8 days at 6 log₁₀ cfu g⁻¹. Additionally, these bacteria decreased the number of filamentous fungi on the leaves. These isolates were found to inhibit the growth of B. cinerea and E. coli O157:H7 in vitro conditions in growing microbiological media. Their efficacy was confirmed in vivo conditions. These isolates inhibited the development of grey mould caused by B. cinerea on lettuce leaves. Two LAB isolates reduced the abundance of the pathogenic bacterium E. coli on spinach leaves by about 0.7 log₁₀ cfu g⁻¹. In glasshouse conditions, LAB stimulated the growth of examined plants. The lactic acid bacteria used in this study showed the capacity to be used as possible alternatives to chemical compounds in the protection of leafy vegetables against grey mould and for a decrease in E. coli O157:H7 contamination. Full article

This study examined the potential use of three bacterial strains—Paraburkholderia sp. strain CRV74, Pseudomonas sp. strain CRV21, and Acinetobacter sp. strain CRV19—as biocontrol agents of Botrytis cinerea in grapevine. These strains were selected for their ability to inhibit B. cinerea growth in vitro and used in field conditions for the control of grey mould symptoms in ‘Glera’ grapes. To this end, after inoculating these microorganisms onto plants sprayed with B. cinerea spores, the final yield, the physicochemical characteristics of the must, disease incidence, and the possible influence on the expression of plant-defence proteins were evaluated. Strain CRV21 resulted as being the most effective in combating grey mould (−20% of disease incidence). Although yield was not affected, significantly different values of total soluble solids content was observed. Additionally, a significant up-regulation of the genes PR-1, PR-5, β-1,3-glucanase, and class III chitinase was observed. These findings highlight the potential application of strains with anti-botrytis activity as sustainable alternatives to chemical defence for the control of this pathogen. Full article

In this paper, issues related to the technology of compound castings composed of two parts, i.e., the working layer and the supporting part, made of X46Cr13 high-chromium steel and EN-GJL-HB 255 grey cast iron, respectively, in a liquid–solid system by pre-installing a monolithic insert in the mould cavity are presented. As a part of the research, the mechanism of formation of transitional zones in the bonding area of the above-mentioned two alloys was identified and described. It was shown that the phenomenon that determines the formation of a permanent bond between the joined materials is the transport of C and heat from the “high-carbon and hot” material of the supporting part poured into the mould in the form of liquid cast iron to the “low-carbon and cold” material of the working layer placed in the form of a steel monolithic insert inside the mould cavity. In the paper, the suitability of the compound castings technology developed for use in the coke industry is also presented. Full-size high-chromium steel–grey cast iron compound casting plates designed for the coke quenching car lining were positively verified in real coke plant operating conditions. Full article

The marketing value of table grapes is contingent upon several quality requirements, mostly related to microbial decay, sugar/acidity ratio, and colour. This research explores the impact of combining organic-cultured compatible techniques to delay disorders along with organic grape distribution in post-harvest. Aurebasidum pullulans in-field application on grape bunches at three growing stages as a biocontrol agent against grey mould growth coupled with massive modified atmosphere packaging (MMAP; 20% CO₂, 10% O₂) equipped with a breathable valve was tested. The in-field treatment had a significant impact on the colour and sugar content of the grapes at harvest and the mould count evolution during storage, whilst the trend of the other parameters was mainly affected by the interaction of the variables tested. The untreated batch experienced the worst behaviour and the packaging was paramount in preserving the moisture content and appearance of the bunches. The findings of this study may contribute to developing novel practices for setting a smart distribution of organic table grapes and reducing food waste. Full article

The use of synthetic sanitizers for fresh commodities preservation is of concern, with eco-friendly alternatives, including essential oils (EOs), attracting research and industry interest. Dittany (Origanum dictamnus—DIT) oil was applied, either through vapour or dipping, on pepper fruit or in vitro against Botrytis cinerea, and compared to untreated (control) or chlorine (CHL)-treated fruits stored at 11 °C. Direct DIT vapours (up to 6 d) suppressed lesion growth, spore germination, and spore production compared to the untreated fruits. The antimicrobial properties of EOs were evidenced in pre-exposed-to-DIT oil vapours (residual effect), resulting in fruit lesion suppression. However, DIT-pretreated fruits had the same spore production and spore germination as the control and CHL applications. In in vitro tests, DIT vapours decreased colony growth and spore production when fungi were grown on Potato Dextrose Agar (PDA) or PDA pre-exposed to DIT following B. cinerea inoculation. This evidenced that the disease suppression after DIT vapour application primarily affected the interaction of the fruit–pathogen and/or residual responses on fruit tissue and/or PDA media. Fungal biomass in Potato Dextrose Broth (PDB) was evaluated after DIT and CHL applications (10–50–100–500–5000 μL L⁻¹) and decreased with the DIT oil. Additionally, DIT or CHL sanitary dipping on pepper was ineffective against B. cinerea compared to vapour application. Therefore, DIT vapours revealed antimicrobial properties and could be an alternative postharvest sanitiser. DIT oil application should also be evaluated at semi-commercial scale for further optimizations, prior to commercialisation. Full article

Botrytis cinerea can infect almost all of the important horticultural crops and cause severe economic losses globally every year. Modifying candidate genes and studying the phenotypic changes are among the most effective ways to unravel the pathogenic mechanism of this crop killer. However, few effective positive selection markers are used for B. cinerea genetic transformation, which limits multiple modifications to the genome, especially genes involving redundant functions. Here, we optimized a geneticin resistance gene, BcNPTII, based on the codon usage preference of B. cinerea. We found that BcNPTII can greatly increase the transformation efficiency of B. cinerea under G418 selection, with approximately 30 times higher efficiency than that of NPTII, which is applied efficiently to transform Magnaporthe oryzae. Using the gene replacement method, we successfully knocked out the second gene BOT2, with BcNPTII as the selection marker, from the mutant ΔoahA, in which OAHA was first replaced by the hygromycin resistance gene HPH in a field strain. We obtained the double knockout mutant ΔoahA Δbot2. Our data show that the codon-optimized BcNPTII is an efficient positive selection marker for B. cinerea transformation and can be used for various genetic manipulations in B. cinerea, including field wild-type strains. Full article