Search Results (44)

In recent years, significant advancements in artificial intelligence, particularly in the field of deep learning, have increasingly been integrated into agricultural applications, including critical processes such as disease detection. Tomato, being one of the most widely consumed agricultural products globally and highly susceptible to a variety of fungal, bacterial, and viral pathogens, remains a prominent focus in disease detection research. In this study, we propose a deep learning-based approach for the detection of tomato diseases, a critical challenge in agriculture due to the crop’s vulnerability to fungal, bacterial, and viral pathogens. We constructed an original dataset comprising 6414 images captured under real production conditions, categorized into three image types: leaves, green tomatoes, and red tomatoes. The dataset includes five classes: healthy samples, late blight, early blight, gray mold, and bacterial cancer. Twenty-one deep learning models were evaluated, and the top five performers (EfficientNet-b0, NasNet-Large, ResNet-50, DenseNet-201, and Places365-GoogLeNet) were selected for feature extraction. From each model, 1000 deep features were extracted, and feature selection was conducted using MRMR, Chi-Square (Chi²), and ReliefF methods. The top 100 features from each selection technique were then used for reclassification with traditional machine learning classifiers under five-fold cross-validation. The highest test accuracy of 92.0% was achieved with EfficientNet-b0 features, Chi² selection, and the Fine KNN classifier. EfficientNet-b0 consistently outperformed other models, while the combination of NasNet-Large and Wide Neural Network yielded the lowest performance. These results demonstrate the effectiveness of combining deep learning-based feature extraction with traditional classifiers and feature selection techniques for robust detection of tomato diseases in real-world agricultural environments. Full article

Trichoderma harzianum, a prominent biocontrol microorganism, often exhibits restricted colonization efficiency in nutrient-poor soil, thus reducing its biocontrol effectiveness. This study investigated the impact of vitamin C industrial fermentation byproduct (residue after evaporation, RAE), which is recognized for enhancing plant growth and stress tolerance, on the colonization ability and anti-pathogenic fungi activity of T. harzianum through in vitro and pot experiments. In vitro experiments demonstrated that RAE and its main component (2-keto-L-gulonic acid, 2KGA) significantly enhanced biomass and spore production (41.44% and 158.46% on average) of two T. harzianum strains in an oligotrophic medium (1/5 PDA). In a more nutrient-limited medium (1/10 PDA), RAE significantly increased the inhibition rates of T. harzianum S against Fusarium graminearum, Botrytis cinerea, and Alternaria alternata by 6.12–7.77%. Pot experiments further revealed that, compared with T. harzianum application alone, the combined application of RAE and T. harzianum S, (1) significantly elevated T. harzianum S abundance by 23.77% while significantly reducing B. cinerea abundance by 33.78% in rhizosphere soil; (2) significantly improved the content of soil available phosphorus (147.63%), ammonium nitrogen (60.05%), and nitrate nitrogen (32.19%); and (3) significantly improved the superoxide dismutase activity (17.39%) and fresh weight of tomato plants (130.74%). Correlation analysis revealed that there were significant positive correlations between T. harzianum S abundances/plant biomass and RAE, and significantly negative correlations between B. cinerea abundance and T. harzianum S/plant biomass/peroxidase activity. Collectively, RAE effectively promoted the growth of T. harzianum and pathogen suppression ability, while improving soil fertility and tomato biomass. This study offers novel insights into RAE’s agricultural application for plant disease control while supporting the sustainable development of vitamin C production. Full article

Phytopathogenic fungi and bacteria pose a serious threat to global agriculture, leading to significant economic losses and potential health risks. Consequently, the search for natural alternatives to synthetic agrochemicals has garnered increasing scientific attention, with plant extracts emerging as promising environmentally friendly solutions. In this context, the surface extract of Salvia discolor, obtained using dichloromethane, was analyzed for its bioactive potential. Chemical profiling revealed a rich composition of terpenoids and flavonoids. The antimicrobial potential of the ground extract was evaluated against nine phytopathogenic fungi (Alternaria solani, Botrytis cinerea, Colletotrichum lindemuthianum, Fusarium solani, Fusarium oxysporum f. sp. lactucae race 1, Phoma betae, Phaeomoniella chlamydospora, Pythium dissotocum, and Stemphylium sp.), and two phytopathogenic bacteria (Clavibacter michiganesis subsp. michiganesis and Pectobacterium carotovorum subsp. carotovorum), selected from common pathogens of agricultural interest. Complete inhibition of P. chlamydospora at 1000 µg mL⁻¹ and strong activity against P. dissotocum, F. solani and B. cinerea was observed, and low inhibition (<40%) against C. lindemuthianum and F. oxysporum f. sp. lactucae race 1. However, the extract showed promising results in the post-harvest protection of tomatoes against gray mold. Moderate antibacterial activity was seen against C. michiganensis subsp. michiganensis. These findings indicate that S. discolor extract has the potential to serve as an effective natural crop protection agent, though further optimization may be needed for broader application. Full article

One novel C₁₀ polyacetylene rhamnoside, 4,6,8-decatriyne-1-O-α-L-rhamnopyranoside, named xylariside A (1), together with two novel C₁₀ polyacetylene quinovopyranosides, 4,6,8-decatriyne-1-O-α-D-quinovopyranoside, xylariside B (2), and 8E-decaene-4,6-diyne-1-O-α-D-quinovopyranoside, xylariside C (3), were obtained from the solid fermentation of Xylaria sp. VDL4, an endophytic fungus isolated from Vaccinium dunalianum wight (Ericaceae). Their chemical structures were elucidated through a combination of spectroscopic techniques. The antifungal activities of these compounds were evaluated in vitro against four phytopathogenic fungi (Fusarium oxysporum, Botrytis cinerea, Phytophthora capsici, and Fusarium solani). Compound 2 demonstrated significant antifungal activities, with minimum inhibitory concentration (MIC) values ranging from 3.91 to 7.81 μg/mL. Compound 2’s effectiveness levels were similar to those of the reference drugs thiabendazole and carbendazim (each MIC = 0.98−15.62 μg/mL). Xylariside B (2) was further evaluated against B. cinerea in vivo. It exhibited remarkable efficacy in both the prevention and treatment of tomato and strawberry gray mold. Molecular docking studies confirmed the antifungal mechanism of compound 2 by revealing its binding interactions with key enzyme targets in B. cinerea, thereby supporting the observed in vitro and in vivo results. Additionally, compound 2 showed effective inhibition of α-glucosidase, with IC₅₀ values of 5.27 ± 0.0125 μg/mL. Full article

Tomato (Solanum lycopersicum L.) is a major economic vegetable crop globally, yet it is prone to gray mold disease caused by Botrytis cinerea infection during cultivation. Caffeoyl shikimate esterase (CSE) is a crucial component of the lignin biosynthesis pathway, which significantly contributes to plant stress resistance. Therefore, investigating the expression patterns of SlCSE after Botrytis cinerea infection may offer a theoretical foundation for breeding resistant tomato varieties. In this study, 11 SlCSE family members were identified from the tomato genome using bioinformatics analyses. Public transcriptome databases and RT-qPCR experiments were used to analyze gene expression in tomato tissues, responses to Botrytis cinerea infection, and the temporal characteristics of the response to 2-ethylfuran treatment during infection. These experiments resulted in the identification of the key gene SlCSE06. Transgenic tomato lines that overexpressed SlCSE06 were constructed to examine their resistance levels to gray mold disease. Many SlCSE genes were upregulated when tomato fruit were infected with Botrytis cinerea during the ripening stage. Furthermore, 24 h after treatment with 2-ethylfuran, most SlCSE genes exhibited increased expression levels compared with the control group, but they exhibited significantly lower levels at other time points. Thus, 2-ethylfuran treatment may enhance the responsiveness of SlCSEs. Based on this research, SlCSE06 was identified as the key gene involved in the response to Botrytis cinerea infection. The SlCSE06-overexpressing (OE6) tomato plants exhibited a 197.94% increase in expression levels compared to the wild type (WT). Furthermore, the lignin content in OE6 was significantly higher than in WT, suggesting that the overexpression of SlCSE06 enhanced lignin formation in tomato plants. At 5 days post-inoculation with Botrytis cinerea, the lesion diameter in OE6 decreased by 31.88% relative to the WT, whereas the lignin content increased by 370.90%. Furthermore, the expression level of SlCSE06 was significantly upregulated, showing a 17.08-fold increase compared with the WT. These findings suggest that 2-ethylfuran enhances the activation of the critical tomato disease resistance gene SlCSE06 in response to gray mold stress, thereby promoting lignin deposition to mitigate further infection by Botrytis cinerea. Full article

Gray mold caused by Botrytis cinerea is a common postharvest fungal disease in fruit and vegetables. The prevention and treatment of postharvest gray mold has been one of the hot research issues addressed by researchers. This study aimed to investigate the effect of L-methionine and L-arginine on Botrytis cinerea in vitro and on cherry tomato fruit. The results of the in vitro experiment showed that L-methionine and L-arginine had significant inhibitory effects on the mycelial growth and spore germination of Botrytis cinerea, and the inhibitory effects were enhanced with increasing L-methionine or L-arginine concentration. In addition, L-methionine and L-arginine treatment increased the leakage of Botrytis cinerea electrolytes, proteins and nucleic acids. The experiment involving propidium iodide staining and malondialdehyde content assay also confirmed that L-methionine and L-arginine treatment could lead to cell membrane rupture and lipid peroxidation. The results of scanning electron microscopy further verified that the morphology of hyphae was damaged, deformed, dented and wrinkled after treatment with L-methionine or L-arginine. Fruit inoculation experiments displayed that L-methionine and L-arginine treatments significantly inhibited the occurrence and development of gray mold in postharvest cherry tomato. Therefore, treatment with L-methionine or L-arginine might be an effective means to control postharvest gray mold in fruit and vegetables. Full article

The most significant aspect of promoting greenhouse productivity is the timely monitoring of disease spores and applying proactive control measures. This paper introduces a method to classify spores of airborne disease in greenhouse crops by using fingerprint characteristics of diffraction–polarized images and machine learning. Initially, a diffraction–polarization imaging system was established, and the diffraction fingerprint images of disease spores were taken in polarization directions of 0°, 45°, 90° and 135°. Subsequently, the diffraction–polarization images were processed, wherein the fingerprint features of the spore diffraction–polarization images were extracted. Finally, a support vector machine (SVM) classification algorithm was used to classify the disease spores. The study’s results indicate that the diffraction–polarization imaging system can capture images of disease spores. Different spores all have their own unique diffraction–polarization fingerprint characteristics. The identification rates of tomato gray mold spores, cucumber downy mold spores and cucumber powdery mildew spores were 96.02%, 94.94% and 96.57%, respectively. The average identification rate of spores was 95.85%. This study can provide a research basis for the identification and classification of disease spores. Full article

In order to reduce the use of fungicide and ensure food safety, it is necessary to develop fungicide with low toxicity and high efficiency to reduce residues. Azoxystrobin (AZOX), which is derived from mushrooms, is an excellent choice. However, conventional AZOX release is difficult to regulate. In this paper, a pH-responsive fungicide delivery system for the preparation of AZOX by impregnation method was reported. The Zinc metal–organic framework/Biomass charcoal (ZIF-8/BC) support was first prepared, and subsequently, the AZOX-ZIF-8/BC nano fungicide was prepared by adsorption of AZOX onto ZIF-8/BC by dipping. Gray mold, caused by Botrytis cinerea, is one of the most important crop diseases worldwide. AZOX-ZIF-8/BC could respond to oxalic acid produced by Botrytis cinerea to release loaded AZOX. When pH = 4.8, it was 48.42% faster than when pH = 8.2. The loading of AZOX on ZIF-8/BC was 19.83%. In vitro and pot experiments showed that AZOX-ZIF-8/BC had significant fungicidal activity, and 300 mg/L concentration of AZOX-ZIF-8-BC could be considered as a safe and effective control of Botrytis cinerea. The above results indicated that the prepared AZOX-ZIF-8/BC not only exhibited good drug efficacy but also demonstrated pH-responsive fungicide release. Full article

Botrytis cinerea, a causal agent of gray mold disease, is one of the most destructive fungal pathogens that leads to substantial global economic crop losses, especially for tomato plants. The present study aims to investigate the inhibitory effect of two microbial culture filtrates (BCA filtrate alone and combined with salicylic acid) of Trichoderma longibrachiatum and Pseudomonas sp. against the phytopathogenic fungus B. cinerea on tomato plants. The biochemical modifications, gray mold disease incidence, and fruit quality parameters of the tomatoes were determined according to tested treatments. The results showed that both fungi and bacteria were able to solubilize phosphate and produce IAA and HCN. T. longibrachiatum could produce hydrolytic enzymes (chitinase, protease, and glucanase). Otherwise, Pseudomonas sp. showed the capacity to produce catalase and amylase enzymes. Both microbial culture filtrates inhibited the hyphae growth of B. cinerea. The biocontrol efficacy, in vitro, was significant: up to 50% in terms of the growth inhibition rate at a concentration of 40%. The tomato seedlings’ growth was promoted by the separate preventive treatments of each micro-organism culture filtrate. In addition, disease severity in the tomato seedlings and fruit was significantly reduced. Furthermore, the combined treatment of tomato fruit with culture filtrates and salicylic acid induced significant biochemical and physiological changes in fruit firmness, juice yield, total protein, and ROS enzyme activities. The culture filtrates of T. longibrachiatum and Pseudomonas sp. can be recommended as an effective microbial biofungicide to control gray mold disease under storage conditions. Full article

The present study provides a comprehensive overview of the Bacillus velezensis strain Htq6, and its potential applications in plant disease control. Htq6 is an endophytic bacterium derived from walnut, which was found to possess a strong inhibitory effect on a wide range of plant pathogenic microorganisms and was identified as a good plant disease control agent. The entire genome of the Bacillus velezensis Htq6 was sequenced, and a comparative genomic analysis was conducted with various Bacillus species in order to better understand the mechanism of the strain’s biological control. At the same time, a new classification result was presented. Additionally, transcriptome analysis was performed to explore the response mechanism of tomato gray mold fungus after treatment with the fermentation liquid of Bacillus velezensis Htq6. The study analyzed the distribution of various secondary metabolite gene clusters in the Bacillus model strains and employed RNA-Seq technology to obtain transcriptome expression profiles. Furthermore, the cell wall, cell membrane, and antioxidant-related genes of Botrytis cinerea were analyzed, providing insight into the antibacterial mechanism of biocontrol bacteria and the stress response mechanism of Botrytis cinerea. The results of the research are promising, and could potentially lead to the development of an effective biocontrol agent for the prevention and control of various plant diseases. Full article

Bacillus mojavensis D50, a biocontrol strain, is used to prevent and treat the fungal plant pathogen Botrytis cinerea. Bacillus mojavensis D50’s biofilms can affect its colonization; thus, the effects of different metal ions and culture conditions on biofilm formation were determined in this study. The results of medium optimization showed that Ca²⁺ had the best ability to promote biofilm formation. The optimal medium composition for the formation of biofilms contained tryptone (10 g/L), CaCl₂ (5.14 g/L), and yeast extract (5.0 g/L), and the optimal fermentation conditions included pH 7, a temperature of 31.4 °C, and a culture time of 51.8 h. We found that the antifungal activity and abilities to form biofilms and colonize roots were improved after optimization. In addition, the levels of expression of the genes luxS, SinR, FlhA, and tasA were up-regulated by 37.56-, 2.87-, 12.46-, and 6.22-fold, respectively. The soil enzymatic activities which related biocontrol-related enzymes were the highest when the soil was treated by strain D50 after optimization. In vivo biocontrol assays indicated that the biocontrol effect of strain D50 after optimization was improved. Full article

Gray mold caused by Botrytis cinerea causes significant losses in tomato crops. B. cinerea infection may be halted by volatile organic compounds (VOCs), which may exhibit fungistatic activity or enhance the defense responses of plants against the pathogen. The enhanced VOC generation was observed in tomato (Solanum lycopersicum L.), with the soil-applied biocontrol agent Trichoderma virens (10⁶ spores/1 g soil), which decreased the gray mold disease index in plant leaves at 72 hpi with B. cinerea suspension (1 × 10⁶ spores/mL). The tomato leaves were found to emit 100 VOCs, annotated and putatively annotated, assigned to six classes by the headspace GCxGC TOF-MS method. In Trichoderma-treated plants with a decreased grey mold disease index, the increased emission or appearance of 2-hexenal, (2E,4E)-2,4-hexadienal, 2-hexyn-1-ol, 3,6,6-trimethyl-2-cyclohexen-1-one, 1-octen-3-ol, 1,5-octadien-3-ol, 2-octenal, octanal, 2-penten-1-ol, (Z)-6-nonenal, prenol, and acetophenone, and 2-hydroxyacetophenone, β-phellandrene, β-myrcene, 2-carene, δ-elemene, and isocaryophyllene, and β-ionone, 2-methyltetrahydrofuran, and 2-ethyl-, and 2-pentylfuran, ethyl, butyl, and hexyl acetate were most noticeable. This is the first report of the VOCs that were released by tomato plants treated with Trichoderma, which may be used in practice against B. cinerea, although this requires further analysis, including the complete identification of VOCs and determination of their potential as agents that are capable of the direct and indirect control of pathogens. Full article

Trichoderma harzianum is a well-known biological control agent (BCA) that is effective against a variety of plant pathogens. In previous studies, we found that T. harzianum T4 could effectively control the gray mold in tomatoes caused by Botrytis cinerea. However, the research on its biocontrol mechanism is not comprehensive, particularly regarding the mechanism of mycoparasitism. In this study, in order to further investigate the mycoparasitism mechanism of T. harzianum T4, transcriptomic sequencing and real-time fluorescence quantitative PCR (RT-qPCR) were used to identify the differentially expressed genes (DEGs) of T. harzianum T4 at 12, 24, 48 and 72 h of growth in the cell wall of B. cinerea (BCCW) or a sucrose medium. A total of 2871 DEGs and 2148 novel genes were detected using transcriptome sequencing. Through GO and KEGG enrichment analysis, we identified genes associated with mycoparasitism at specific time periods, such as encoding kinases, signal transduction proteins, carbohydrate active enzymes, hydrolytic enzymes, transporters, antioxidant enzymes, secondary metabolite synthesis, resistance proteins, detoxification genes and genes associated with extended hyphal longevity. To validate the transcriptome data, RT-qCPR was performed on the transcriptome samples. The RT-qPCR results show that the expression trend of the genes was consistent with the RNA-Seq data. In order to validate the screened genes associated with mycoparasitism, we performed a dual-culture antagonism test on T. harzianum and B. cinerea. The results of the dual-culture RT-qPCR showed that 15 of the 24 genes were upregulated during and after contact between T. harzianum T4 and B. cinerea (the same as BCCW), which further confirmed that these genes were involved in the mycoparasitism of T. harzianum T4. In conclusion, the transcriptome data provided in this study will not only improve the annotation information of gene models in T. harzianum T4 genome, but also provide important transcriptome information regarding the process of mycoparasitism at specific time periods, which can help us to further understand the mechanism of mycoparasitism, thus providing a potential molecular target for T. harzianum T4 as a biological control agent. Full article

Owing to the remarkable antimicrobial potential of these materials, research into the possible use of nanomaterials as alternatives to fungicides in sustainable agriculture is increasingly progressing. Here, we investigated the potential antifungal properties of chitosan-decorated copper oxide nanocomposite (CH@CuO NPs) to control gray mold diseases of tomato caused by Botrytis cinerea throughout in vitro and in vivo trials. The nanocomposite CH@CuO NPs were chemically prepared, and size and shape were determined using Transmission Electron Microscope (TEM). The chemical functional groups responsible for the interaction of the CH NPs with the CuO NPs were detected using the Fourier Transform Infrared (FTIR) spectrophotometry. The TEM images confirmed that CH NPs have a thin and semitransparent network shape, while CuO NPs were spherically shaped. Furthermore, the nanocomposite CH@CuO NPs ex-habited an irregular shape. The size of CH NPs, CuO NPs and CH@CuO NPs as measured through TEM, were approximately 18.28 ± 2.4 nm, 19.34 ± 2.1 nm, and 32.74 ± 2.3 nm, respectively. The antifungal activity of CH@CuO NPs was tested at three concentrations of 50, 100 and 250 mg/L and the fungicide Teldor 50% SC was applied at recommended dose 1.5 mL/L. In vitro experiments revealed that CH@CuO NPs at different concentrations significantly inhibited the reproductive growth process of B. cinerea by suppressing the development of hyphae, spore germination and formation of sclerotia. Interestingly, a significant control efficacy of CH@CuO NPs against tomato gray mold was observed particularly at concentrations 100 and 250 mg/L on both detached leaves (100%) as well as the whole tomato plants (100%) when compared to the conventional chemical fungicide Teldor 50% SC (97%). In addition, the tested concentration 100 mg/L improved to be sufficient to guarantee a complete reduction in the disease’s severity (100%) to tomato fruits from gray mold without any morphological toxicity. In comparison, tomato plants treated with the recommended dose 1.5 mL/L of Teldor 50% SC ensured disease reduction up to 80%. Conclusively, this research enhances the concept of agro-nanotechnology by presenting how a nano materials-based fungicide could be used to protect tomato plants from gray mold under greenhouse conditions and during the postharvest stage. Full article

Botrytis cinerea, the causal agent of gray mold, is one of the most destructive pathogens of cherry tomatoes, causing fruit decay and economic loss. Fludioxonil is an effective fungicide widely used for crop protection and is effective against tomato gray mold. The emergence of fungicide-resistant strains has made the control of B. cinerea more difficult. While the genome of B. cinerea is available, there are few reports regarding the large-scale functional annotation of the genome using expressed genes derived from transcriptomes, and the mechanism(s) underlying such fludioxonil resistance remain unclear. The present study prepared RNA-sequencing (RNA-seq) libraries for three B. cinerea strains (two highly resistant (LR and FR) versus one highly sensitive (S) to fludioxonil), with and without fludioxonil treatment, to identify fludioxonil responsive genes that associated to fungicide resistance. Functional enrichment analysis identified nine resistance related DEGs in the fludioxonil-induced LR and FR transcriptome that were simultaneously up-regulated, and seven resistance related DEGs down-regulated. These included adenosine triphosphate (ATP)-binding cassette (ABC) transporter-encoding genes, major facilitator superfamily (MFS) transporter-encoding genes, and the high-osmolarity glycerol (HOG) pathway homologues or related genes. The expression patterns of twelve out of the sixteen fludioxonil-responsive genes, obtained from the RNA-sequence data sets, were validated using quantitative real-time PCR (qRT-PCR). Based on RNA-sequence analysis, it was found that hybrid histidine kinase, fungal HHKs, such as BOS1, BcHHK2, and BcHHK17, probably involved in the fludioxonil resistance of B. cinerea, in addition, a number of ABC and MFS transporter genes that were not reported before, such as BcATRO, BMR1, BMR3, BcNMT1, BcAMF1, BcTOP1, BcVBA2, and BcYHK8, were differentially expressed in the fludioxonil-resistant strains, indicating that overexpression of these efflux transporters located in the plasma membranes may associate with the fludioxonil resistance mechanism of B. cinerea. All together, these lines of evidence allowed us to draw a general portrait of the anti-fludioxonil mechanisms for B. cinerea, and the assembled and annotated transcriptome data provide valuable genomic resources for further study of the molecular mechanisms of B. cinerea resistance to fludioxonil. Full article