Inhibition Of Spore Germination Research Articles

Ebselen improves fungal keratitis through exerting anti-inflammation, anti-oxidative stress, and antifungal effects

Fungal keratitis is a severely vision-threatening corneal infection, where the prognosis depends on both fungal virulence and host immune defense. Inappropriate host responses can induce substantial inflammatory damage to the cornea. Therefore, in the treatment of fungal keratitis, it is important to concurrently regulate the immune response while efforts are made to eliminate the pathogen. Ebselen is a widely studied organo-selenium compound and has been demonstrated to have antifungal, antibacterial, anti-inflammatory, and oxidative stress-regulatory properties. The effectiveness of ebselen for the treatment of fungal keratitis remains unknown. In this study, ebselen was demonstrated to produce a marked inhibitory effect on Aspergillus fumigatus (A. fumigatus), including spore germination inhibition, mycelial growth reduction, and fungal biofilm disruption. The antifungal activity of ebselen was related to the cell membrane damage caused by thioredoxin (Trx) system inhibition-mediated oxidative stress. On the contrary, ebselen enhanced the antioxidation of Trx system in mammalian cells. Further, ebselen was proven to suppress the expressions of inflammatory mediators (IL-1β, IL-6, TNF-α, COX-2, iNOS, and CCL2) and reduce the production of oxidative stress-associated indicators (ROS, NO, and MDA) in fungi-stimulated RAW264.7 cells. In addition, ebselen regulated PI3K/Akt/Nrf2 and p38 MAPK signaling pathways, which contributed to the improvement of inflammation and oxidative stress. Finally, we verified the therapeutic effect of ebselen on mouse fungal keratitis. Ebselen improved the prognosis and reduced the fungal burden in mouse corneas. Expressions of inflammatory mediators, as well as the infiltration of macrophages and neutrophils in the cornea were also obviously decreased by ebselen. In summary, ebselen exerted therapeutic effects by reducing fungal load and protecting host tissues in fungal keratitis, making it a promising treatment for fungal infections.

Screening of Endophytic Antagonistic Bacteria in Wheat and Evaluation of Biocontrol Potential against Wheat Stripe Rust.

Wheat stripe rust is globally one of the most important diseases affecting wheat. There is an urgent need to develop environmentally safe and durable biological control options to supplement the control that is achieved with breeding and fungicides. In this study, endophytic bacteria were isolated from healthy wheat through the tissue separation method. Antagonistic endophytic bacteria were screened based on the control effect of urediniospore germination and wheat stripe rust (WSR). The taxonomic status of antagonistic strains was determined based on morphological, physiological, and biochemical characteristics and molecular biological identification (16S rDNA and gyrB gene sequence analysis). Finally, the potential growth-promoting effect of different concentrations of antagonists on wheat seedlings and the biological control effect of WSR were studied. A total of 136 strains of endophytic bacteria belonging to 38 genera were isolated. Pseudomonas was the most common bacterial genus, with 29 isolates (21%). The biological control effect of different isolates was assessed using an urediniospore germination assay. The isolate XD29-G1 of Paenibacillus polymyxa had the best performance, with 85% inhibition of spore germination during primary screening. In the deep screening, the control effect of XD29-G1 on wheat stripe rust was 60%. The antagonist XD29-G1 promoted the germination of wheat seeds and the growth of wheat seedlings at a solution dilution of 10-7 cfu/mL. The pot experiment results showed that different dilution concentrations of the strain had different levels of antibacterial activity against WSR, with the concentration of 10-1 cfu/mL having the best control effect and a control efficiency of 61.19%. XD29-G1 has better biological control potential against wheat stripe rust.

Sanxiapeptin is an ideal preservative with a dual effect of controlling green mold and inducing systemic defense in postharvest citrus

Green mold is a common postharvest disease infected by Penicillium digitatum that causes citrus fruit decay, and severely affects fruit storage quality. This work aimed to investigate the antifungal activity of Sanxiapeptin against P. digitatum, and elucidate the possible mechanisms involved. Sanxiapeptin was capable of inhibiting spore germination, germ tube length and mycelial growth. The SYTOX green staining assay revealed that Sanxiapeptin targeted the fungal membrane, and changed the membrane permeability, leading to the leakage of cell constituents. Meanwhile, Sanxiapeptin could influence the cell wall permeability and integrity by increasing the activities of chitinase and glucanase, resulting in abnormal chitin consumption and the decrease of glucan. Intriguingly, Sanxiapeptin could effectively control postharvest decay in citrus fruits, and activate the host resistance responses by regulating the phenylpropanoid pathway. In conclusion, Sanxiapeptin exhibits multiphasic antifungal mechanisms of action to control green mold in citrus fruits, shows great potential as novel food preservatives.

Synthesis, Characterization, and Antifungal Activity of Benzimidazole-Grafted Chitosan against Aspergillus flavus.

Aspergillus flavus contamination in agriculture and food industries poses threats to human health, leading to a requirement of a safe and effective method to control fungal contamination. Chitosan-based nitrogen-containing derivatives have attracted much attention due to their safety and enhanced antimicrobial applications. Herein, a new benzimidazole-grafted chitosan (BAC) was synthesized by linking the chitosan (CS) with a simple benzimidazole compound, 2-benzimidazolepropionic acid (BA). The characterization of BAC was confirmed by Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance spectroscopy (1H and 13C NMR). Then, the efficiency of BAC against A. flavus ACCC 32656 was investigated in terms of spore germination, mycelial growth, and aflatoxin production. BAC showed a much better antifungal effect than CS and BA. The minimum inhibitory concentration (MIC) value was 1.25 mg/mL for BAC, while the highest solubility of CS (16.0 mg/mL) or BA (4.0 mg/mL) could not completely inhibit the growth of A. flavus. Furthermore, results showed that BAC inhibited spore germination and elongation by affecting ergosterol biosynthesis and the cell membrane integrity, leading to the permeabilization of the plasma membrane and leakage of intracellular content. The production of aflatoxin was also inhibited when treated with BAC. These findings indicate that benzimidazole-derived natural CS has the potential to be used as an ideal antifungal agent for food preservation.

Gα3 regulates growth and development, stress response, patulin synthesis and pathogenicity of Penicillium expansum by mediating cAMP/PKA and PKC signaling

G proteins are signaling proteins in eukaryotes that transduce extracellular signals into the intracellular space. There are three classes of Gα proteins in fungi, among which class III Gα proteins play crucial roles for fungal growth, secondary metabolism, and pathogenicity. In this study, we found that the Gα3 protein from Penicillium expansum belongs to the P-loop NTPase superfamily and contains a G-alpha conserved domain and an adenylate cyclase binding site that can bind to guanine nucleotides. Deletion of Gα3 downregulated the expression of the cAMP/PKA signaling pathway genes AC, PkaC, and PkaR, and the PKC signaling related genes PKC1, PKC2, and PLC of P. expansum. The Gα3-deleted strain showed slow colony growth, irregular depression at the colony edge, and increased branching at the mycelial tip. Gα3 deletion decreased spore production, inhibited spore germination, and reduced the expression levels of genes that regulate conidiospore formation, including abaA, wetA, vosA and brlA. In addition, the Gα3-deleted strain grew faster in carbon and NO3- medium, and was more sensitive to salt and oxidative stress, but less sensitive to cell wall stress. Gα3 deletion significantly downregulated the expression of patulin biosynthesis-related genes except for PatF and PatM, and reduced the patulin levels in vivo and in vitro. In addition, loss of Gα3 reduced the pathogenicity of the strain on fruit, with down-regulated expression of the extracellular enzyme genes PG, PL, and PLA. In conclusion, Gα3 can regulate growth and development, carbon and nitrogen response, stress response, patulin biosynthesis and pathogenicity by mediating cAMP/PKA and PKC signaling pathways of P. expansum.

Characterization of a broad-spectrum antifungal strain, Streptomyces graminearus STR-1, against Magnaporthe oryzae.

Fungal diseases such as the devastating rice blast pose severe threats to crop production worldwide. Biological control of crop diseases caused by fungal pathogens is an environment-friendly approach for safeguarding crop production. But the insufficient availability of microbial agents effective against various fungal diseases has hampered the development of green production in crops. In this study, we identified a broad-spectrum antifungal bacterium, Streptomyces graminearus STR-1, showing antagonistic activity to diverse fungal pathogens including Magnaporthe oryzae, Rhizoctonia solani, Fusarium graminearum, Ustilaginoidea virens, and Bipolaris maydis. Its antifungal activity was relatively stable and less affected by temperature and pH. Evaluation of the biocontrol activity of STR-1 revealed that STR-1 prevented and controlled rice blast disease via eliciting plant immunity and suppressing fungal infection-structure development. STR-1 broth extract inhibited spore germination, likely through inhibiting protein synthesis. Combining LC-MS and chromatography analysis of the antimicrobial compounds purified from STR-1 broth extract, together with decoding STR-1 genomic sequence, we identified 4-oxo-4-[(1-phenylethyl)amino]but-2-enoic acid, 1,3,5-Trimethylpyrazole and SMA-1 as the potential main STR-1 secondary metabolites associated with its antifungal effects. This study suggests that bacterial strain STR-1 could be used for identifying highly effective and broad-spectrum secondary metabolites for containing rice blast and other crop diseases. The application of the active compounds offers a promising measure to tackle fungal disease.

Antifungal efficacy of chitosan extracted from shrimp shell on strawberry (Fragaria × ananassa) postharvest spoilage fungi

The strong demand for biological materials in the food industry places chitosan at the forefront of other biopolymers. The present study aims to evaluate the antifungal properties of chitosan extracted from shrimp shell waste (Parapenaeus longirostris) against post-harvest strawberry (Fragaria × ananassa) spoilage fungi. The physicochemical characteristics (DD, Mw, and solubility) of extracted chitosan were determined. In addition, functional characteristics were studied by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The antifungal effect of chitosan on mycelial growth and spore germination of Aspergillus niger, Botrytis cinerea, Fusarium oxysporum, and Rhizopus stolonifer was evaluated. Yield, degree of deacetylation, molecular weight, and solubility were 21.86%, 83.50%, 180 kDa, and 80.10%, respectively. A degree of deacetylation of 81.27% was calculated from the FTIR spectrum and a crystallinity index of 79.83% was determined from the X-ray diffraction pattern. SEM images of extracted chitosan showed a combination of fibrous and porous structure. At 3% chitosan, mycelial growth inhibition rates of A. niger, B. cinerea, F. oxysporum, and R. stolonifer ranged from 81.37% to 92.70%. At the same chitosan concentration, the percentages of spore germination inhibition of the isolated fungi ranged from 65.47% to 71.48%. The antifungal activity was highly dose-dependent. As a natural polymer, chitosan offers a convincing alternative to synthetic antimicrobials for the post-harvest preservation of strawberries. Its potential lies in its ability to inhibit the growth of spoilage fungi.

The rice endophytic bacterium Bacillus velezensis LS123N provides protection against multiple pathogens and enhances rice resistance to wind with increase in yield

Rice (Oryza sativa) is a globally important staple food, but its production was influenced by various biotic and abiotic stresses. This study involved screening endophytic bacterial strains for their antagonistic activities and control efficacy against multiple rice pathogens in planta assays. Subsequent greenhouse investigations identified Bacillus velezensis LS123N as the most promising strain for effectively controlling various rice diseases, including bacterial blight, rice blast, bakanae disease, Pythium disease, and brown spot disease. Field trials further demonstrated that LS123N significantly reduced naturally occurring brown spot disease caused by Bipolaris oryzae in both seedlings and mature rice plants, and increased rice yields. For disease control against bacterial blight, Pythium disease, and brown spot, only a single treatment during seed germination resulted in significant disease control efficacy. The application of LS123N during seed germination aligns with common rice cultivation practices. Additionally, LS123N exhibited the capacity to improve rice resistance to wind stress and enhance rice growth during the seedling stage. Endophytic colonization of LS123N was evidenced from seed to mature rice plants after seed treatment. Interestingly, the expression of many rice defense genes was suppressed within 48 h of the LS123N introduction. In vitro assays showed that LS123N inhibited spore germination and normal growth of germ tubes of B. oryzae, produced multiple hydrolases, siderophores, and IAA, and had the ability to solubilize phosphorus compounds. In summary, these results indicate that B. velezensis LS123N is a promising biocontrol agent and has the potential to be developed into a biofungicide, biofertilizer, and biostimulator.

Molecular identification and management of mycotoxigenic fungi in stored corn Grains

AbstractMycotoxin-producing molds which considered as common maize grains contaminants are the genera Fusarium, Aspergillus and Penicillium. There are natural and safe ways to protect grains from mold contamination as the use of essential oils and chemical treatments. A total number of 25 samples were used to study the natural frequency in five governorates in Egypt, Molecular identification indicated that the most frequent fungi were Fusarium verticillioides, Aspergillus niger, Talaromyces verruculosus, Aspergillus flavus and Aspergillus terreus. The in vitro studies have been done to determine mycelial growth and spore germination inhibition of the two A. flavus; isolated and reference isolates. Thyme and acetic acid were tested in direct contact assay to study their effects on mycelial growth. Treatments showed significant impact on mycelial growth and spore germination inhibition of both A. flavus isolates. In the postharvest application treatments: as vapour and carrier contact assay, Thyme and Acetic acid were tested to determine their influence on growth and aflatoxin production in A. flavus isolates by liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS). Results indicated that both treatments were effective in inhibition of aflatoxin production in both vapour and carrier assays as they succeeded in reducing AFB1 while they inhibited completely the production of AFB2. The extent of the inhibition of aflatoxin production was dependent on the concentration and storage duration of treatments applied.

Antifungal evaluation of turmeric rhizome extract against Colletotrichum capsici, the causal agent of anthracnose on red-chili peppers (Capsicum annum L.)

This study was conducted to evaluate the antifungal activity of the ethanolic extract of turmeric rhizome against Colletotrichum capsici Syd. and its efficacy in reducing anthracnose disease development on red chili peppers. The in vitro experiment aimed to determine the effect of the extract on inhibiting C. capsici growth in potato sucrose agar (PSA) medium incorporating the turmeric rhizome extract. The variables examined were mycelial growth, fungal sporulation, and spore germination. The in vivo experiment aimed to evaluate the effectiveness of the extract in reducing disease development on red chili peppers. The treatments were conducted using a completely randomized design with five concentration treatments and three replicates. The variables examined were radial mycelial growth zone diameter, fungal sporulation, spore germination, and disease intensity. Data were analyzed by analysis of variance, and the least significant difference was used for comparing means between treatments. The results of these experiments showed that ethanolic turmeric rhizome extract exhibited antifungal activity against C. capsici both in vitro and in vivo conditions. Turmeric rhizome extracts with concentrations of 1%, 2%, 3%, and 4% significantly inhibited mycelial growth, and even at a 4% concentration, C. capsici fungal colonies did not grow. Evaluation of the effect of the extract on fungal sporulation showed that inhibition occurred at a concentration of 3%, while inhibition of spore germination occurred at concentrations of 1%, 2%, and 3%. Application of turmeric extract at concentrations of 1%, 2%, 3%, and 4% effectively reduced the intensity of anthracnose on red chili.

Attenuation of phytofungal pathogenicity of Ascomycota by autophagy modulators

Autophagy in eukaryotes functions to maintain homeostasis by degradation and recycling of long-lived and unwanted cellular materials. Autophagy plays important roles in pathogenicity of various fungal pathogens, suggesting that autophagy is a novel target for development of antifungal compounds. Here, we describe bioluminescence resonance energy transfer (BRET)-based high-throughput screening (HTS) strategy to identify compounds that inhibit fungal ATG4 cysteine protease-mediated cleavage of ATG8 that is critical for autophagosome formation. We identified ebselen (EB) and its analogs ebselen oxide (EO) and 2-(4-methylphenyl)−1,2-benzisothiazol-3(2H)-one (PT) as inhibitors of fungal pathogens Botrytis cinerea and Magnaporthe oryzae ATG4-mediated ATG8 processing. The EB and its analogs inhibit spore germination, hyphal development, and appressorium formation in Ascomycota pathogens, B. cinerea, M. oryzae, Sclerotinia sclerotiorum and Monilinia fructicola. Treatment with EB and its analogs significantly reduced fungal pathogenicity. Our findings provide molecular insights to develop the next generation of antifungal compounds by targeting autophagy in important fungal pathogens.

Evaluation of the Most Effective Plant Extracts, Bioagents and Chemical Fungicides In vitro against Powdery Mildew of Pea

The present investigation was carried during 2022-23 at Plant Pathology Section, College of Agriculture, Pune-05 with the objectives of evaluation of the most effective plant extracts, bioagents and chemical fungicides in vitro. Effect of six plant extracts, bioagents and chemical fungicides were studied in vitro by hanging drop method where the study revealed that garlic (Allium sativum), followed by neem (Azadiracta indica) showed highest inhibition of spore germination for powdery mildew. After 48 hours of incubation, the spore germination rate ranged from 3.15 % to 34.45 %, where garlic showed the highest inhibition at 95.30 %, followed by neem at 91.03 %. The efficacy of bioagents was evaluated by preparing the bacterial and fungal culture filtrates where Trichoderma harzianum and Pseudomonas fluorescens were found to be the most effective with tune of 56.86% and 50.72% of efficacy respectively against Erysiphe pisi. Among all chemical fungicides, hexaconazole (0.05 %) resulted as the most effective chemical fungicide against powdery mildew of pea. These findings can be useful in developing safer and more eco-friendly methods to control powdery mildew and protect crops.

In vitro Antifungal Efficacy of Some Plant Extracts against Fungal Pathogens Causing Diseases in Solanaceous Crops

Background: This research aimed to assess the antifungal potential of oil and organic leaf extracts obtained from Ocimum gratissimum, Kopsia fruticosa and Hibiscus rosa-sinensis against fungal pathogens affecting solanaceous crops. Methods: Oil of O. gratissimum was obtained by hydro-distilled method using Clevenger apparatus. Leaf extracts of K. fruticosa and H. rosa-sinensis were extracted using three organic solvents namely hexane, ethanol and methanol. The antifungal activity of the oil and organic leaf extracts against the test fungal pathogens was determined by the agar well diffusion method. The efficacy of the oil and organic leaf extracts against the pathogens was evaluated by their minimum inhibitory concentration (MIC), ability to inhibit spore germination and growth kinetic assay. Result: The oil of O. gratissimum (1000 ppm) significantly inhibited the growth of Cercospora capsici (81.4%), Alternaria alternata (73.7%) and Fusarium oxysporum (68.7%). Hexane extract of K. fruticosa and methanol extract of H. rosa-sinensis (1500 µg/well) displayed strong antifungal activity against Colletotrichum acutatum with inhibition percentages of 75.9% and 82.4% respectively. Similarly, ethanol extract of K. fruticosa and methanol extract of H. rosa-sinensis showed effective inhibition against C. capsici and A. alternata. The oil of O. gratissimum showed MIC values of 50, 100 and 200 µg/mL against C. capsici, A. alternata and F. oxysporum respectively. Ethanol extract of K. fruticosa showed MIC values ranging from 100 to 350 µg/mL, while H. rosa-sinensis methanol extract displayed MIC values ranging from 50 to 200 µg/mL against the fungal pathogens. The oil extract also demonstrated strong inhibition of spore germination of the test fungi. Our study suggests the potential of plant-derived essential oils and extracts as effective antifungal agents for crop protection.

Antifungal Effect of Metabolites from Bacterial Symbionts of Entomopathogenic Nematodes on Fusarium Head Blight of Wheat.

Fungal diseases such as Fusarium head blight (FHB) are significant biotic stressors, negatively affecting wheat production and quality. This study explored the antifungal activity of the metabolites produced by the bacterial symbionts of entomopathogenic nematodes (EPNs) against FHB-causing Fusarium sp. Fusarium graminearum. To achieve this, the symbiotic bacteria of nine EPN isolates from the EPN collection at the Agricultural Research Council-Small Grains (ARC-SG) were isolated from the cadavers of Galleria mellonella (Lepidoptera: Pyralidae) larvae after infection with EPNs. Broth cultures (crude) and their supernatants (filtered and autoclaved) of each bacterial isolate were used as bacterial metabolite treatments to test their inhibitory effect on the mycelial growth and spore germination of F. graminearum. Mycelial growth inhibition rates varied among both bacterial isolates and treatments. Crude metabolite treatments proved to be more effective than filtered and autoclaved metabolite treatments, with an overall inhibition rate of 75.25% compared to 23.93% and 13.32%, respectively. From the crude metabolite treatments, the Xenorhabdus khoisanae SGI 197 bacterial isolate from Steinernema beitlechemi SGI 197 had the highest mean inhibition rate of 96.25%, followed by Photorhabdus luminescens SGI 170 bacteria isolated from Heterorhabditis bacteriophora SGI 170 with a 95.79% mean inhibition rate. The filtered metabolite treatments of all bacterial isolates were tested for their inhibitory activity against Fusarium graminearum spore germination. Mean spore germination inhibition rates from Xenorhabdus spp. bacterial isolates were higher (83.91 to 96.29%) than those from Photorhabdus spp. (6.05 to 14.74%). The results obtained from this study suggest that EPN symbiotic bacterial metabolites have potential use as biological control agents of FHB. Although field efficacy against FHB was not studied, the significant inhibition of mycelial growth and spore germination suggest that the application of these metabolites at the flowering stage may provide protection to plants against infection with or spread of F. graminearum. These metabolites have the potential to be employed as part of integrated pest management (IPM) to inhibit/delay conidia germination until the anthesis (flowering stage) of wheat seedlings has passed.

Phytochemical characterization of forest leaves extracts and application to control apple postharvest diseases

The study investigated the antifungal and phytochemical properties of three forest plants (Eucalyptus globulus, Pistacia lentiscus, and Juniperus phoenicea) against apple diseases caused by Colletotrichum gloeosporioides and Alternaria alternata. The determination of the total polyphenol and flavonoid contents in the three aqueous extracts of studied plants showed that E. globulus exhibited the highest contents than those of P. lentiscus and J. phoenicea. Furthermore, the three studied extracts showed very appreciable antioxidant activity with decreasing order: E. globulus, P. lentiscus, and J. phoenicea. The phytochemical analysis showed different common phenolic acids in the three studied plants namely: quinic acid, gallic acid, chlorogenic acid, and caffeoylquinic acid as well as other flavonoids mainly quercetin and catechin. The results of the current study demonstrated that the fungistatic activity of E. globulus EO (4 and 2 µl/ml) seemed to be the most effective under laboratory conditions with an inhibition zone diameter above 16 mm. However, the poisoned food technique indicated that the aqueous extract (80%) and the essential oil (4 µl/ml) of E. globulus exhibited the highest mycelial growth (> 67%) and spore germination (> 99%) inhibition. Preventive treatments with essential oils (4 µl/ml) and aqueous extracts (80%) applied to apple fruits inoculated with A. alternata and C. gloeosporioides resulted in the lowest lesion diameter (< 6.80 mm) and disease severity index (< 15%) and the most favorable inhibitory growth (> 85.45%) and protective potentials (> 84.92%). The results suggest that E. globulus has a brilliant future in the management of anthracnose and Alternaria rot of apple and provide a basis for further studies on its effects under field conditions.

Bacillus velezensis BE2 controls wheat and barley diseases by direct antagonism and induced systemic resistance.

Wheat and barley rank among the main crops cultivated on a global scale, providing the essential nutritional foundation for both humans and animals. Nevertheless, these crops are vulnerable to several fungal diseases, such as Septoria tritici blotch and net blotch, which significantly reduce yields by adversely affecting leaves and grain quality. To mitigate the effect of these diseases, chemical fungicides have proven to be genuinely effective; however, they impose a serious environmental burden. Currently, biocontrol agents have attracted attention as a sustainable alternative to fungicides, offering an eco-friendly option. The study aimed to assess the efficacy of Bacillus velezensis BE2 in reducing disease symptoms caused by Zymoseptoria tritici and Pyrenophora teres. This bacterium exhibited significant antagonistic effects in vitro by suppressing fungal development when pathogens and the beneficial strain were in direct confrontation. These findings were subsequently confirmed through microscopic analysis, which illustrated the strain's capacity to inhibit spore germination and mycelial growth in both pathogens. Additionally, the study analysed the cell-free supernatant of the bacterium using UPLC-MS (ultra-performance liquid chromatography-mass spectrometry). The results revealed that strain BE2 produces, among other metabolites, different families of cyclic lipopeptides that may be involved in biocontrol. Furthermore, the beneficial effects of strain BE2 in planta were assessed by quantifying the fungal DNA content directly at the leaf level after bacterization, using two different application methods (foliar and drenching). The results indicated that applying the beneficial bacterium at the root level significantly reduced pathogens pressure. Finally, gene expression analysis of different markers showed that BE2 application induced a priming effect within the first hours after infection. KEY POINTS: • BE2 managed Z. tritici and P. teres by direct antagonism and induced systemic resistance. • Strain BE2 produced seven metabolite families, including three cyclic lipopeptides. • Application of strain BE2 at the root level triggered plant defense mechanisms.

Recruiting grape‐isolated antagonistic yeasts for the sustainable bio‐management of Botrytis cinerea on grapes

AbstractBotrytis cinerea is the causative agent of grey mould disease in grapes, which was linked to significant postharvest losses. This study examined three grape‐isolated yeasts (Metschnikowia aff. fructicola, Metschnikowia pulcherrima, and Hansenispora uvarum) through in vitro and in vivo tests on detached grape berries against grey mould, as well as the elucidation of their possible mechanisms of action. The antifungal mechanism of action of yeasts was determined by the lytic enzyme activity, inhibition of spore germination, biofilm activity, iron depletion, diffusible metabolites, wound‐site colonisation, mycocin, and volatile organic compounds (VOCs) production. The highest in vitro efficacy (83.13%) was observed on M. aff. fructicola, followed by M. pulcherrima (82.10%) and H. uvarum (71.66%). Metschnikowia yeasts exhibited comparable enzyme activities, including protease, β‐1,3 glucanase, gelatinase chitinase, and cellulase, while H. uvarum had a poor enzymatic activity with chitinase and gelatinase. M. aff. fructicola showed relatively higher iron depletion activity than M. pulcherrima, while M. pulcherrima outperformed via diffusible metabolites. All yeast cultures significantly reduced spore germination by at least 86%. Overall, M. aff. fructicola exhibited the highest biocontrol activity with its iron depletion, inhibition of conidial germination, biofilm formation, VOCs, and well colonisation on grape berries. M. aff. fructicola 1‐UDM outperformed all other yeasts by significantly reducing disease incidence and lesion diameter values (93.4% and 94.3%, respectively). Remarkably, H. uvarum VOCs demonstrated potential as a biofumigant for suppressing grey mould. All yeasts are well adapted to their ecological niche to bio‐protect grapes from grey mould disease.

Activity of fungicide cyclobutrifluram against Fusarium fujikuroi and mechanism of the pathogen resistance associated with point mutations in FfSdhB, FfSdhC2 and FfSdhD

Rice bakanae disease (RBD) is a devastating plant disease caused by Fusarium fujikuroi. This study aimed to evaluate the potential of cyclobutrifluram, a novel succinate dehydrogenase inhibitor (SDHI), to control RBD, and determine the risk and mechanism of resistance to cyclobutrifluram in F. fujikuroi. In vitro experiments showed that cyclobutrifluram significantly inhibited mycelial growth and spore germination, and altered the morphology of mycelia and conidia. Treatment with cyclobutrifluram significantly decreased mycotoxin production and increased cell membrane permeability in F. fujikuroi. The baseline sensitivity of 72 F. fujikuroi isolates to cyclobutrifluram was determined using mycelial growth and spore germination inhibition assays, which revealed EC50 values of 0.0114 – 0.1304 μg mL-1 and 0.0012 – 0.016 μg mL-1, with mean EC50 values of 0.0410 ± 0.0470 μg mL-1 and 0.0038 ± 0.0015 μg mL-1, respectively. Pot experiments demonstrated that the protective effect of cyclobutrifluram against F. fujikuroi was more significant than that of phenamacril and azoxystrobin, indicating that cyclobutrifluram is a promising antifungal agent for the control of RBD. Six cyclobutrifluram-resistant mutants of F. fujikuroi were obtained via fungicide adaptation. Moreover, these mutants exhibited weaker fitness than their parental isolate and positive cross-resistance with other SDHI fungicides, including pydiflumetofen and penflufen; however, no cross-resistance was detected with other classes of fungicides, including phenamacril, fludioxonil, prochloraz, or azoxystrobin. These results indicated that the resistance risk of F. fujikuroi to cyclobutrifluram might be moderate. Sequencing analysis revealed that mutations, including H248D in FfSdhB, A83V in FfSdhC2, and S106F and E166K in FfSdhD, contributed to resistance, which was confirmed by molecular docking and homologous replacement experiments. The results suggest a high potential for cyclobutrifluram to control RBD and a moderate resistance risk of F. fujikuroi to cyclobutrifluram, which are meaningful findings for the scientific application of cyclobutrifluram.

Anti-fungal activity of a novel succinate dehydrogenase inhibitor pydiflumetofen against Bipolaris maydis

Southern corn leaf blight (SCLB) is a disease caused by Bipolaris maydis. Pydiflumetofen is a novel succinate dehydrogenase inhibitor (SDHI) fungicide. However, its inhibitory activity against B. maydis has not been investigated in depth. In this study, mycelial growth rate and spore germination methods were used to establish a baseline for the sensitivity of 102 B. maydis strains collected from five regions of China to pydiflumetofen. The results showed that pydiflumetofen exhibited strong inhibitory activity against B. maydis. The EC50 values ranged from 0.0292 to 0.5207 μg/mL and from 0.0115 to 0.4883 μg/mL, with the average EC50 values of 0.2050 ± 0.1779 μg/mL and 0.1858 ± 0.0994 μg/mL for inhibition of mycelial growth and spore germination, respectively. After treatment with pydiflumetofen, the apical branching of mycelia increased significantly, and morphological changes such as twisting, deformation, rupture, and shriveling of mycelia and spores were observed. In addition, it significantly increased cell membrane permeability and decreased succinate dehydrogenase (SDH) activity and adenosine triphosphate (ATP) content. Moreover, the SDH gene expressions were changed, in which MbSdhB, MbSdhC and MbSdhD expressions were significantly upregulated, except for MbSdhA expression, which was significantly downregulated. In detached leaf experiments, pydiflumetofen showed strong protective and curative effects against B. maydis, superior to the same concentration of prochloraz and pyraclostrobin, and its protective effect was better than its curative effect. The field trials showed that the control efficacy of pydiflumetofen at 225 g/hm2 was >70% and its control efficacy at 300 g/hm2 reached 81.77%, which was significantly higher than that of pyraclostrobin and epoxiconazole at the same dosage. After 1:1 mixing of pydiflumetofen with the above control agents, the control efficacies of these mixtures at the same dosage increased compared to the use of a single agent. All the above results demonstrated that pydiflumetofen exhibits excellent inhibitory activity against B. maydis and good control of SCLB, suggesting that it could be used as an alternative fungicide for the control of SCLB.

Effects of zinc oxide nanocomposites on microorganism growth and protection of physicochemical quality during maize storage

Maize moldy and spoilage due to microbial growth is a significant challenge in grain storage. This study aimed to evaluate the effectiveness of a zinc oxide nanocomposite, ZnO@mSiO2, prepared in our previous research, in inhibiting mold growth and preserving maize cell quality. The results demonstrated that ZnO@mSiO2 could effectively inhibit the growth of dominant microorganism, Aspergillus flavus, Talaromyces variabilis, Penicillium citrinum and Fusarium graminearum, in maize storage. Aspergillus flavus was selected as the model fungus, ZnO@mSiO2 effectively disrupted fungal hyphae structure, leading to reduced hyphal mass and inhibited spore germination. The inhibitory effect of ZnO@mSiO2 on mold growth was concentration-dependent. However, the ZnO@mSiO2 at an appropriate concentration (not exceeding 3.0 g/kg) preserved the integrity of maize cell membranes and enhancing the antioxidant activity within maize cells. The findings highlight the potential of ZnO@mSiO2 as an effective protectant to inhibit mold growth and preserve maize quality during storage.