Search

Late leaf spot of groundnut is caused by the fungal pathogen of Cercosporidium personatum. Efforts to prevent this disease generally use synthetic fungicides that can cause environmental pollution. This study aims to identify the fungal pathogen of groundnut late leaf spot and study the antifungal effect of leaf extract of some plants for leaf spot intensity suppression. Late leaf spot pathogen was identified based on morphological characteristics of hyphae and conidia. The leaf extracts of butterfly pea (Clitoria ternatea), Mexican sunflower (Tithonia diversifolia), basil (Ocimum gratissimum), and sugar apple (Annona squamosa) at a concentration of 25% against C. personatum were evaluated in vitro based on growth inhibition. The growth and intensity of groundnut disease attacks were assessed for seven weeks by inoculating C. personatum and application of the leaf extracts. Results showed that the pathogen was identified as C. personatum with conidia characteristics of cylindrical to obclavate, short and slightly curved with four septa, and 35.75 × 7.11 µm in size. The percentage of inhibition of C. personatum in vitro reached 94.03% by leaf extract of C. ternatea, 88.56% by T. diversifolia, 87.20% by O. gratissimum, and 84.10% by A. squamosa. Treatment of C. ternatea increased plant height up to 12.85% and reduced the most optimal percentage of the infected leaves up to 61.53%. C. ternatea leaf extract was found to be the most effective extract to suppress C. personatum infection by 77.94%, while T. diversifolia, O. gratissimum, and A. squamosa reduced disease severity by 70.15%, 65.43%, and 57.76%, respectively.

Ganoderma boninense , the causative agent behind basal stem rot (BSR) disease in oil palm, instigated a pressing need for innovative and ecologically sustainable strategies to counter its impact. Biological control strategies, particularly employing bio-fungicides, have emerged as environmentally friendly alternatives for managing oil palm plant diseases. This study delved into harnessing the biocontrol potential of Indigenous bacterial isolate by investigating and evaluating its bioactive compounds of ethyl acetate extract with the capacity to suppress G. boninense growth in oil palm plants. Molecular identification was utilized to identify of bacterial isolate using 16S rRNA primers (5′- AGA GTT TGA TCC TGG CTC AG– 3′) and (5′- GGA TAC CTT GTT ACG ACT T– 3′). The agar well diffusion technique and scanning electron microscopy (SEM) were utilized to assess the inhibitory effects of bacterial ethyl acetate extract against G. boninense. A comprehensive analysis of the bacterial ethyl acetate extracts was conducted by gas chromatography - mass spectrometry (GC–MS). Molecular identification confirmed that the bacterial isolate was Bacterium strain BS1727. By employing SEM, the ethyl acetate extracts had influenced on G. boninense mycelial architecture. The exposure to the bacterial ethyl acetate extracts induced degradation and morphological distortion of the mycelial structure, serving as an indicator of its impact on the mycelium's structural modulation. Noteworthy, the spectrum of bioactive compounds was a subset of aromatic hydrocarbons, encompassing volatile organic compounds (VOCs). Based on GC–MS analysis, among the VOCs, acetic acid butyl ester and toluene showed the highest relative peak areas, 46.34 % for acetic acid butyl ester and 31.82 % for toluene, respectively. Ethylbenzene had a relative peak area of 5.4 %, 1,1,2-trichloroethane 4.41 %, and allyldimethyl (prop-1-ynyl) silane 3.17 %. The remaining compounds were 6-undecyl-5,6-dihydro-2H-pyran-2-one, 3-decen-5-one, 2-methyl-, and o-cymene, with the relative peak areas were below 1 %. These findings explained the inherent capacity of antifungal compounds as a viable strategy in inhibiting the growth of G. boninense. [ABSTRACT FROM AUTHOR]

Aims Anthracnose caused by Colletotrichum species is one of the most devastating diseases of fruits and crops. We isolated and identified an antifungal compound from the mushroom Coprinus comatus and investigated its inhibitory potential against anthracnose disease-causing fungi with the goal of discovering natural products that can suppress anthracnose-caused plant disease. Methods and results The culture filtrate of C. comatus was subjected to a bioassay-guided isolation of antifungal compounds. The active compound was identified as orsellinaldehyde (2,4-dihydroxy-6-methylbenzaldehyde) based on mass spectroscopy and nuclear magnetic resonance analyses. Orsellinaldehyde displayed broad-spectrum inhibitory activity against different plant pathogenic fungi. Among the tested Colletotrichum species, it exhibited the lowest IC50 values on conidial germination and germ tube elongation of Colletotrichum orbiculare. The compound also showed remarkable inhibitory activity against Colletotrichum gloeosporiodes. The staining of Colletotrichum conidia with fluorescein diacetate and propidium iodide demonstrated that the compound is fungicidal. The postharvest in - vivo detached fruit assay indicated that orsellinaldehyde suppressed anthracnose lesion symptoms on mango and cucumber fruits caused by C. gloeosporioides and C. orbiculare , respectively. Conclusions Orsellinaldehyde was identified as a potent antifungal compound from the culture filtrate of C. comatus. The inhibitory and fungicidal activities of orsellinaldehyde against different Colletotrichum species indicate its potential as a fungicide for protecting various fruits against anthracnose disease-causing fungi. [ABSTRACT FROM AUTHOR]

BACKGROUND: Fungal phytopathogens are a significant threat to crops and food security, and there is a constant need to develop safe and effective compounds that antagonize them. In‐planta assays are complex and tedious and are thus not suitable for initial high‐throughput screening of new candidate antifungal compounds. We propose an in vitro screening pipeline that integrates five rapid quantitative and qualitative methods to estimate the efficacy and mode of action of prospective antifungal compounds. RESULTS: The pipeline was evaluated using five documented antifungal compounds (benomyl, catechol, cycloheximide, 2,4‐diacetylphloroglucinol, and phenylacetic acid) that have different modes of action and efficacy, against the model soilborne fungal pathogen Fusarium oxysporum f. sp. radicis cucumerinum. We initially evaluated the five compounds' ability to inhibit fungal growth and metabolic activity using green fluorescent protein (GFP)‐labeled F. oxysporum and PrestoBlue staining, respectively, in multiwell plate assays. We tested the compounds' inhibition of both conidial germination and hyphal elongation. We then employed FUN‐1 and SYTO9/propidium iodide staining, coupled to confocal microscopy, to differentiate between fungal growth inhibition and death at the cellular level. Finally, using a reactive oxygen species (ROS)‐detection assay, we were able to quantify ROS production in response to compound application. CONCLUSIONS: Collectively, the proposed pipeline provides a wide array of quantitative and qualitative data on the tested compounds that can help pinpoint promising novel compounds; these can then be evaluated more vigorously using in planta screening assays. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Abstract Cytospora canker has become a devastating disease of apple species worldwide, and in severe cases, it may cause dieback of entire trees. The aim of this study was to characterize the diversity of cultivable bacteria from the wild apple microbiota and to determine their antifungal ability against the canker-causing pathogenic fungi Cytospora mali and C. parasitica. Five bacterial strains belonging to the species Bacillus amyloliquefaciens, B. atrophaeus, B. methylotrophicus, B. mojavensis, and Pseudomonas synxantha showed strong antagonistic effects against pathogenic fungi. Therefore, since the abovementioned Bacillus species produce known antifungal compounds, we characterized the antifungal compounds produced by Ps. synxantha. Bacteria grown on nutritional liquid medium were dehydrated, and the active compound from the crude extract was isolated and analysed via a range of chromatographic processes. High-performance liquid chromatography, mass spectrometry, and nuclear magnetic resonance analyses revealed a bioactive antifungal compound, phenazine-1-carboxylic acid (PCA). The minimum inhibitory concentration (MIC) demonstrated that PCA inhibited mycelial growth, with a MIC of 10 mg mL−1. The results suggested that PCA could be used as a potential compound to control C. mali and C. malicola, and it is a potential alternative for postharvest control of canker disease.

Late leaf spot of groundnuts is caused by the fungal pathogen Cercosporidium personatum. Efforts to prevent this disease generally use synthetic fungicides that can pollute the environment. This study aims to identify the fungal pathogen of groundnut late leaf spot and determine the antifungal effect of leaf extract from several plants on leaf spot disease suppression. Late leaf spot pathogen was identified based on hyphae and conidia morphological characteristics. Growth inhibition of leaf extracts from butterfly pea (Clitoria ternatea), Mexican sunflower (Tithonia diversifolia), basil (Ocimum gratissimum), and sugar apple (Annona squamosa) at aconcentration of 25%were tested against C. personatum in vitro. The growth and intensity of groundnut disease attackswere assessed for seven weeks by inoculating C. personatum and applied with several leaf extracts. Results showed that the pathogen was identified as C. personatum with conidia characteristics of cylindrical to obclavate, short and slightly curved with four septa, and 35.75 × 7.11 µm in size. The inhibition percentage of C. personatum in vitro reached 94.03% by leaf extract of C. ternatea, 88.56% by T. diversifolia, 87.20% by O. gratissimum, and 84.10% by A. squamosa. Treatment of C. ternatea increased plant height by up to 12.85% and showed the highest reduction of infected leaves by up to 61.53%. C. ternatea leaf extract was found to be the most effective leaf extract to suppress C. personatum infection by 77.94%, while T. diversifolia, O. gratissimum, and A. squamosa reduced disease severity by 70.15%, 65.43%, and 57.76%, respectively. [ABSTRACT FROM AUTHOR]

Cucumber anthracnose, caused by Colletotrichum orbiculare, is controlled using fungicides, but the fungus can develop resistance. We previously showed that an ethyl acetate extract of the culture filtrate of Cercospora sp. ME202 (ME202-ECF) has fungicidal activity and inhibits the formation of anthracnose lesions caused by C. orbiculare, but we did not identity the antimicrobial compound(s) or analyze the potential to suppress disease progression. Here, we isolated and identified antifungal compounds produced by Cercospora sp. ME202 and evaluated their antifungal activity against C. orbiculare. We purified the active compound using column chromatography and high-performance liquid chromatography, identified it as traversianal, a tricyclic diterpenoid aldehyde with a molecular weight of 316.4 from ME202-ECF, then determined its structure using liquid chromatography-mass spectrometry with electrospray ionization and nuclear magnetic resonance spectroscopy. In a thin layer chromatography bioassay at Rf 0.92, the isolated traversianal inhibited mycelial growth. Traversianal also inhibited conidial germination of C. orbiculare at 2–10 ppm and had fungicidal activity against conidia at 5–10 ppm after 24 h treatment. Pretreatment of cucumber leaves with 10, 20, or 30 ppm traversianal significantly suppressed lesion formation by C. orbiculare without phytotoxicity. Thus, traversianal produced by Cercospora sp. ME202 has potential as a new fungicide against C. orbiculare. [ABSTRACT FROM AUTHOR]

Aim This study aims to incorporate alginate microparticles containing berberine and fluconazole into two different types of pharmaceutical formulations, to subsequently evaluate the antifungal activity against Candida albicans. Methods and Results Alginate microparticles containing BBR (berberine) and FLU (fluconazole) were produced by the spray-drying technique, characterized and incorporated in two pharmaceutical formulations, a vaginal cream and artificial saliva. Broth microdilution, checkerboard, time–kill curve, and scanning electron microscopy were carried out to determine the antifungal effects of BBR and FLU against C. albicans. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of free BBR were 125 μg ml−1. Synergism between BBR and FLU was demonstrated by a fractional inhibitory concentration index (FICI) = 0.0762. The time–kill curve for the combination BBR + FLU showed a more pronounced decrease in fungal growth in comparison to free drugs, and an antibiofilm effect of BBR occurred in the formation and preformed biofilm. Conclusion Alginate microparticles containing BBR and FLU were obtained and incorporated in a vaginal cream and artificial saliva. Both formulations showed good stability, antifungal effects, and organoleptic characteristics, which suggest that BBR–FLU microparticles in formulations have potential as antifungal therapy. [ABSTRACT FROM AUTHOR]

Aims: Microbial biocontrol agents have become an effective option to mitigate the harmfulness of chemical pesticides in recent years. This study demonstrates the control efficacy of Bacillus velezensis CE 100 on the anthracnose causal agent, Colletotrichum gloeosporioides. Methods and results: In vitro antifungal assays revealed that the culture filtrate and volatile organic compounds of B. velezensis CE 100 strongly restricted the mycelial development of C. gloeosporioides. Moreover, a bioactive compound, butyl succinate, was isolated from the n-butanol crude extract of B. velezensis CE 100 (BCE), and identified by liquid chromatography-electrospray ionization hybrid ion-trap and time-of-flight mass spectrometry (LC-ESI-QTOF-MS) and one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR). Treatment with purified butyl succinate at a concentration of 300 μg mL−1 strongly controlled conidial germination of C. gloeosporioides with an inhibition rate of 98.66%, whereas butyl succinate at a concentration of 400 μg mL−1 showed weak antifungal action on the mycelial growth of C. gloeosporioides with an inhibition rate of 31.25%. Scanning electron microscopy revealed that the morphologies of butyl succinate-treated hyphae and conidia of C. gloeosporioides were severely deformed with shriveled and wrinkled surfaces. Furthermore, butyl succinate was able to control carbendazim-resistant C. gloeosporioides, demonstrating that it could be a promising agent for the suppression of other carbendazim-resistant fungal pathogens. An in vivo biocontrol assay demonstrated that the strain CE 100 broth culture and butyl succinate showed higher control efficacy on apple anthracnose than BCE. Conclusions: Our findings provide insight into the antifungal potential of B. velezensis CE 100 and its butyl succinate for efficient control of phytopathogenic fungi, such as C. gloeosporiodes, in plant disease protection. This is the first study to demonstrate the antifungal potential of bacteria-derived butyl succinate for control of C. gloeosporioides. [ABSTRACT FROM AUTHOR]

Plant pathogens are responsible for important damages to valuable crops causing important economic losses. Agrobiodiversity protection is crucial for the valorization of local varieties that could possess higher resistance to biotic and abiotic stress. At the beginning of germination, seeds are susceptible to pathogens attacks, thus they can release endogenous antimicrobial compounds of different natures in the spermosphere, to contrast proliferation of microorganisms. The work aimed at characterizing the maize of local variety Nostrano di Storo seed exudates secreted during the first phases of germination, to identify compounds active in the defense towards pathogens. Storo seed exudates were proven to inhibit F. verticilloides germination. In order to investigate the cause of the described effect, compositional profiling of the exudates was performed through NMR, lipidomic, and proteomic analyses. This study suggests an important role of microbial endophytic communities in the protection of the seed during the early phases of the germination process and their interplay with fatty acids released by the seeds, rather than a specific antifungal compound. The valorization of agronomically acceptable maize lines with pre-harvest enhanced resistances to pathogens contamination could lead, in the near future, to commercially available varieties potentially requiring more limited chemical protective treatments. [ABSTRACT FROM AUTHOR]

Candida albicans is an opportunistic yeast accounting for about 50–90 % of all cases of candidiasis in humans, ranging from superficial to systemic potentially life-threatening infections. The presence of several virulence factors, including biofilm, hyphal transition, and proteolytic enzymes production, worsens the fungal infections burden on healthcare system resources. Hence, developing new bioactive compounds with antifungal activity is a pressing urgence for the scientific community. In this perspective, we evaluated the anti- Candida potential of the N-Nitroso-N-phenylhydroxylamine ammonium salt (cupferron) against standard and clinical C. albicans strains. Firstly, the in vitro cytotoxicity of cupferron was checked in the range 400–12.5 μg/mL against human microglial cells (HMC-3). Secondly, its antifungal spectrum was explored via disk diffusion test, broth-microdilution method, and time-killing curve analysis, validating the obtained results through scanning electron microscopy (SEM) observations. Additionally, we evaluated the cupferron impact on the main virulence determinants of Candida albicans. At non-toxic concentrations (100–12.5 μg/mL), the compound exerted interesting anti- Candida activity, registering a minimum inhibitory concentration (MIC) between 50 and 100 μg/mL against the tested strains, with a fungistatic effect until 100 μg/mL. Furthermore, cupferron was able to counteract fungal virulence at MIC and sub-MIC values (50–12.5 μg/mL). These findings may propose cupferron as a new potential antifungal option for the treatment of Candida albicans infections. • Antifungal evaluation of cupferron against planktonic and sessile Candida albicans. • Reduction of biofilm, hyphal transition and proteolytic enzymes production. • MIC at 50 μg/mL, with a fungistatic effect until 100 μg/mL. • The cytotoxicity in the range 100–12.5 μg/mL against HMC-3. [ABSTRACT FROM AUTHOR]

Here we report the discovery and identification of antibacterial substances from the secondary metabolites of a new strain Bacillus velezensis BA-26. The whole genome of B. velezensis BA-26 was sequenced. Its genome data were annotated, and four potentially unknown gene clusters and nine known secondary metabolite synthetic gene clusters were analyzed and excavated by antiSMASH software. Based on the predicted secondary metabolites from the genome of strain BA-26, the material identification of isolated and purified extracellular secondary metabolites was conducted in combination with mass spectrometry data. A total of 24 antifungal compounds were identified, namely, iturin and various fengycins. The minimum inhibitory concentration (MIC) of iturin to Botrytis cinerea was 62.50 µg·mL−1. The MICs of C14 fengycin A, C16 fengycin A and C18 fengycin A were 62.50, 31.25 and 0.49 µg·mL−1, respectively, indicating that the increase in the number of carbon atoms in side chain fatty acids of fengycin improves its biological activity. Two kinds of anti-disease fungal compounds from B. velezensis BA-26, namely, iturin and fengycin, were purified and identified; they were found to have antibacterial activities against common pathogenic fungi. Therefore, B. velezensis BA-26 may be potentially used as a biological control agent. This study enriches the genome information on B. velezensis, elucidates the active components of B. velezensis BA-26 antibacterial substances, and provides a useful reference for using strain BA-26 as a biological control agent. Supplemental data for this article is available online at https://doi.org/10.1080/13102818.2021.1934540 .

Bacillus siamensis LDR was tested for its potential as a biocontrol agent against the phytopathogenic fungi Curvularia lunata BM and Ganoderma sp. TB4. Fermentation of B. siamensis LDR for the production of antifungal compound was performed in modified Czapex- Dox broth using hanjeli (Coix lacryma-jobi L.) starch as carbon source. The Bacillus siamensis LDR inoculum was 105 CFU/ mL, and fermentation was conducted for up to 16 days. Antibiosis assay conducted to test the antifungal activity of filtrate medium. The results showed inhibition of C. lunata BM and Ganoderma sp. TB4 were 47.08% and 85.99%, respectively on 14th day of fermentation. Antifungal assay of the crude extract from filtrate medium revealed growth inhibition of C. lunata BM (60.70%) and Ganoderma sp. TB4 (65.25%). Thin layer chromatography of the crude extract revealed pink-colored spots indicative of lipopeptide compounds. Analysis of the crude extract by ultraperformance liquid chromatography-mass spectrometry was tentatively identified as iturin A, bacillomycin F, and surfactin. [ABSTRACT FROM AUTHOR]

Abstract Background “Bayoud” disease caused by Fusarium oxysporum f. sp. albedinis (Foa) poses a serious threat to date palm (Phoenix dactylifera L.) in Morocco. However, research studies performed to discover biological methods to control this disease remain limited. The present study has set objectives to determine antifungal activity of five plants extracts (Acacia cyanophylla, Cupressus atlantica, Eucalyptus torquata, Nerium oleander, and Schinus molle) against Foa and link this effect to their content in polyphenols and flavonoids as well as their antioxidant properties. Results Plant extracts showed significant differences (p < 0.05) regarding their antifungal activity. The extracts of E. torquata and C. atlantica showed the strongest antifungal effect resulting in the inhibition of mycelial growth, sporulation, and spore germination in a dose-dependent manner. In addition, there were significant differences among the examined plant extracts in respect to their total polyphenols (1.536–7.348 g GAE/100 g DW), flavonoids (0.986–5.759 g RE/100 g DW), and antioxidant properties measured by Trolox equivalent antioxidant capacity (TEAC) (7.47–38.97 mmol TE/100 g DW) and ferric-reducing antioxidant power (FRAP) assay (8.95–47.36 mmol TE/100 g DW). Moreover, the antifungal potential of plant extracts was found to be moderately to strongly correlated with their polyphenol and flavonoid contents as well as their antioxidant activity, implying that the effective inhibitory activity of these plant extracts is partly due to their richness in antioxidative secondary metabolites. Conclusion Our findings shed further light on plants as a-yet-untapped resource of bioactive compounds and constructed the foundation for the development of new biological approaches to best manage Bayoud disease.

Fusarium wilt of tomato, a disease caused by the soilborne fungus Fusarium oxysporum f. sp. lycopersici, causes major losses to tomato production. Chemical fungicides and resistant cultivars are the main control strategies, but new races of the pathogen appear to be able to overcome resistance genes in currently grown cultivars. Therefore, there is a need to identify natural antifungal compounds and develop new agents to control F. oxysporum f. sp. lycopersici. The culture filtrate (F31D-CF) from isolate F31D, a bacterial isolate collected from paddy field soil in Shimane Prefecture in 2019, significantly inhibited in vitro conidial germination of F. oxysporum f. sp. lycopersici, indicating a fungicidal effect against this pathogen. The amount of the inhibitory compound in F31D-CF increased from 1 to 3 days of incubation. F31D-CF significantly suppressed disease development in tomato plants. Sequence analysis of the 16S rDNA region of the isolate revealed 99% similarity with the type strain of Streptomyces plumbeus. Thin layer chromatography–bioautography of F31D-CF showed that the compound inhibiting F. oxysporum f. sp. lycopersici growth had an Rf of 0.57. The effective compound in F31D-CF has a molecular weight of > 3000 and is heat stable below 100 °C. Our results suggest that secondary metabolites of isolate F31D have potential for developing a new fungicidal agent against F. oxysporum f. sp. lycopersici. [ABSTRACT FROM AUTHOR]

The gut microflora of insects plays important roles throughout their lives. Different foods and geographic locations change gut bacterial communities. The invasive wood-borer Agrilus mali causes extensive mortality of wild apple, Malus sieversii , which is considered a progenitor of all cultivated apples, in Tianshan forests. Recent analysis showed that the gut microbiota of larvae collected from Tianshan forests showed rich bacterial diversity but the absence of fungal species. In this study, we explored the antagonistic ability of the gut bacteria to address this absence of fungi in the larval gut. The results demonstrated that the gut bacteria were able to selectively inhibit wild apple tree-associated fungi. Among them, Pseudomonas synxantha showed strong antagonistic ability, producing antifungal compounds. Using different analytical methods, such as column chromatography, mass spectrometry, HPLC, and NMR, an antifungal compound, phenazine-1-carboxylic acid (PCA), was identified. Activity of the compound was determined by the minimum inhibitory concentration method and electron microscopy. Moreover, our study showed that the gut bacteria could originate from noninfested apple microflora during infestation. Overall, the results showed that in newly invaded locations, A. mali larvae changed their gut microbiota and adopted new gut bacteria that prevented fungal colonization in the gut. [ABSTRACT FROM AUTHOR]

The gut microflora of insects plays important roles throughout their lives. Different foods and geographic locations change gut bacterial communities. The invasive wood-borer Agrilus mali causes extensive mortality of wild apple, Malus sieversii, which is considered a progenitor of all cultivated apples, in Tianshan forests. Recent analysis showed that the gut microbiota of larvae collected from Tianshan forests showed rich bacterial diversity but the absence of fungal species. In this study, we explored the antagonistic ability of the gut bacteria to address this absence of fungi in the larval gut. The results demonstrated that the gut bacteria were able to selectively inhibit wild apple tree-associated fungi. Among them, Pseudomonas synxantha showed strong antagonistic ability, producing antifungal compounds. Using different analytical methods, such as column chromatography, mass spectrometry, HPLC, and NMR, an antifungal compound, phenazine-1-carboxylic acid (PCA), was identified. Activity of the compound was determined by the minimum inhibitory concentration method and electron microscopy. Moreover, our study showed that the gut bacteria could originate from noninfested apple microflora during infestation. Overall, the results showed that in newly invaded locations, A. mali larvae changed their gut microbiota and adopted new gut bacteria that prevented fungal colonization in the gut.

Botrytis cinerea, the causal agent of gray mold is one of the major devastating fungal pathogens that occurs in strawberry cultivation and leads to massive losses. Due to the rapid emergence of resistant strains in recent years, an ecofriendly disease management strategy needs to be developed to control this aggressive pathogen. Bacillus velezensis CE 100 exhibited strong antagonistic activity with 53.05% against B. cinerea by dual culture method. In the present study, 50% of culture filtrate supplemented into PDA medium absolutely inhibited mycelial growth of B. cinerea whereas the highest concentration (960 mg/L) of different crude extracts including ethyl acetate, chloroform, and n-butanol crude extracts of B. velezensis CE 100, strongly inhibited mycelial growth of B. cinerea with the highest inhibition of 79.26%, 70.21% and 69.59% respectively, resulting in severe damage to hyphal structures with bulging and swellings. Hence, the antifungal compound responsible was progressively separated from ethyl acetate crude extract using medium pressure liquid chromatography. The purified compound was identified as methyl hippurate by nuclear magnetic resonance and mass spectrometry. The inhibitory effect of methyl hippurate on both spore germination and mycelial growth of B. cinerea was revealed by its dose-dependent pattern. The spore germination rate was completely restricted at a concentration of 3 mg/mL of methyl hippurate whereas no mycelial growth was observed in agar medium supplemented with 4 mg/mL and 6 mg/mL of methyl hippurate by poisoned food method. Microscopic imaging revealed that the morphologies of spores were severely altered by long-time exposure to methyl hippurate at concentrations of 1 mg/mL, 2 mg/mL and 3 mg/mL and hyphae of B. cinerea were severely deformed by exposure to methyl hippurate at concentrations of 2 mg/mL, 4 mg/mL and 6 mg/mL. No significant inhibition on tomato seed germination was observed in treatments with methyl hippurate (2 mg/mL) for both 6 h and 12 h soaking period as compared to the controls. Based on these results, B. velezensis CE 100 could be considered a potential agent for development of environmentally friendly disease control strategies as a consequence of the synergetic interactions of diverse crude metabolites and methyl hippurate. [ABSTRACT FROM AUTHOR]

In the search for antifungal agents from marine resources, we recently found that the culture filtrate of Trichoderma longibrachiatum SFC100166 effectively suppressed the development of tomato gray mold, rice blast, and tomato late blight. The culture filtrate was then successively extracted with ethyl acetate and n-butanol to identify the fungicidal metabolites. Consequently, a new compound, spirosorbicillinol D (1), and a new natural compound, 20,30 -dihydro-epoxysorbicillinol (2), together with 11 known compounds (3–13), were obtained from the solvent extracts. The chemical structures were determined by spectroscopic analyses and comparison with literature values. The results of the in vitro antifungal assay showed that of the tested fungal pathogens, Phytophthora infestans was the fungus most sensitive to the isolated compounds, with MIC values ranging from 6.3 to 400 µg/mL, except for trichotetronine (9) and trichodimerol (10). When tomato plants were treated with the representative compounds (4, 6, 7, and 11), bisvertinolone (6) strongly reduced the development of tomato late blight disease compared to the untreated control. Taken together, our results revealed that the culture filtrate of T. longibrachiatum SFC100166 and its metabolites could be useful sources for the development of new natural agents to control late blight caused by P. infestans. [ABSTRACT FROM AUTHOR]

Here we report the discovery and identification of antibacterial substances from the secondary metabolites of a new strain Bacillus velezensis BA-26. The whole genome of B. velezensis BA-26 was sequenced. Its genome data were annotated, and four potentially unknown gene clusters and nine known secondary metabolite synthetic gene clusters were analyzed and excavated by antiSMASH software. Based on the predicted secondary metabolites from the genome of strain BA-26, the material identification of isolated and purified extracellular secondary metabolites was conducted in combination with mass spectrometry data. A total of 24 antifungal compounds were identified, namely, iturin and various fengycins. The minimum inhibitory concentration (MIC) of iturin to Botrytis cinerea was 62.50 µg·mL−1. The MICs of C14 fengycin A, C16 fengycin A and C18 fengycin A were 62.50, 31.25 and 0.49 µg·mL−1, respectively, indicating that the increase in the number of carbon atoms in side chain fatty acids of fengycin improves its biological activity. Two kinds of anti-disease fungal compounds from B. velezensis BA-26, namely, iturin and fengycin, were purified and identified; they were found to have antibacterial activities against common pathogenic fungi. Therefore, B. velezensis BA-26 may be potentially used as a biological control agent. This study enriches the genome information on B. velezensis, elucidates the active components of B. velezensis BA-26 antibacterial substances, and provides a useful reference for using strain BA-26 as a biological control agent. Supplemental data for this article is available online at https://doi.org/10.1080/13102818.2021.1934540. [ABSTRACT FROM AUTHOR]

Rice blast, a devastating disease caused by the fungus Pyricularia oryzae, causes major losses to rice production, but it continuously evolves resistance against fungicides. Therefore, natural compounds need to be identified to develop new agents to control fungal pathogens. An ethyl acetate extract of a culture filtrate (S712-ECF) from isolate S712, a member of Polyporaceae collected from a paddy field in Shimane Prefecture in 2009, was sprayed on rice leaves 24 h before inoculation with conidia of P. oryzae. S712-ECF significantly suppressed blast lesion formation on leaves. Analysis of the internal transcribed spacer (ITS)-5.8S rDNA sequence of S712 showed that it shared high similarities with species of the fungus Annulohypoxylon. S712-ECF significantly inhibited in vitro conidial germination of P. oryzae in a dose-dependent manner and had a fungicidal effect against P. oryzae. Growth inhibition zones of P. oryzae were detected in thin layer chromatography assays of S712-ECF. The most effective compound in S712-ECF was heat-stable with a molecular weight < 3000 Da. Secondary metabolites of isolate S712 thus have potential for developing a new fungicidal agent against P. oryzae. [ABSTRACT FROM AUTHOR]

Information on microbial genome sequences is a powerful resource for accessing natural products with significant activities. We herein report the unveiling of lucensomycin production by Streptomyces achromogenes subsp. streptozoticus NBRC14001 based on the genome sequence of the strain. The genome sequence of strain NBRC14001 revealed the presence of a type I polyketide synthase gene cluster with similarities to a biosynthetic gene cluster for natamycin, which is a polyene macrolide antibiotic that exhibits antifungal activity. Therefore, we investigated whether strain NBRC14001 produces antifungal compound(s) and revealed that an extract from the strain inhibited the growth of Candida albicans. A HPLC analysis of a purified compound exhibiting antifungal activity against C. albicans showed that the compound differed from natamycin. Based on HR-ESI-MS spectrometry and a PubChem database search, the compound was predicted to be lucensomycin, which is a tetraene macrolide antibiotic, and this prediction was supported by the results of a MS/MS analysis. Furthermore, the type I polyketide synthase gene cluster in strain NBRC14001 corresponded well to lucesomycin biosynthetic gene cluster (lcm) in S. cyanogenus, which was very recently reported. Therefore, we concluded that the antifungal compound produced by strain NBRC14001 is lucensomycin.

Following preliminary bioactivity testing, the fungal strain identified as Penicillium crysogenum was cultured in a modified Czapec Yeast Broth medium (CYB) for the production of antifungal compounds. Several chromatographic techniques including HPLC were used to purify the fungal metabolites from the crude extract. The mass determination of the purified compound was performed using Water's LCMS system while the structure of the compound was elucidated using 400 and 500 Varian NMR machines. The chemical name of the purified compound is (2 R, 4S) -2, 4-dimethyl-4-((E)-2-((3S, 4S)-2, 4, 5-trihydroxy-3-methoxy-4-phenyl -1 , 2, 3, 4-tetrahydroquinolin-6-yl) vinyl) cyclohexanone with the chemical formula C 26 H 31 NO 5 and exact mass of 437.2. Molecular docking predicted compound docking score with dihydrofolate reductase enzyme and lanosterol 14α-demethylase enzyme as -8.1 kcal/mol and -9.8 kcal/mol respectively. Further, the compounds showed stable binding mode with the enzymes and reported robust binding energies. After insilico analysis, the compound with mass 437 was tested for its antifungal potential in vitro against two pathogenic yeast species (i.e. Candida albicans and Candida glaberata) using the agar tube diffusion method. Using sterile di-methyl sulfoxide (DMSO) the compound was prepared in four dose concentrations (100, 250, 500, 1000 µg mL -1 ) and mixed with autoclaved semisolid Potato Dextrose Agar (PDA) medium in screw-capped test tubes labelled with the corresponding dose concentration. The fungal strains were inoculated on this medium and linear growth inhibition of the fungal strains was calculated using fluconazole as the control drug. The results from in vitro experiments were encouraging as at concentrations of 500 and 1000 μg mL -1 the compound inhibited the growth of C. albicans by 17% and 38% while 19% and 41% inhibition were recorded against C. glaberata . The compound showed antifungal activity in silico and in vitro against both the Candida species and can act as a potent antifungal candidate in the future upon further investigation.Communicated by Ramaswamy H. Sarma.

Humoral defenses are the major components of insect innate immune system that include the production of several soluble effector molecules from fat body and hemocytes, and released in to the hemolymph upon microbial infection. Hemolymph was collected from the fungal immunized fifth instar larvae of tasar silkworm, Antheraea mylitta , extracted with a mixture of solvent (methanol/glacial acetic acid/water) and fractionated through RP-HPLC. Several fractions were collected, lyophilized and their antifungal activity was tested against Candida albicans. Only the fraction showing strong antifungal activity was further purified via gel filtration chromatography and the purity of active compound was confirmed by thin layer chromatography which showed only single spot after staining with ninhydrin. The molecular mass of this purified compound was determined by high resolution mass spectrometry as 531 Da and analysis of 1H and 13C NMR spectral data along with mass fragmentation pattern indicated the probable structure of the isolated compound as symmetric bis-decanoate derivative. Scanning electron microscopic study revealed that the compound degraded fungal cell wall leading to its lysis and may be the major target for its antifungal activity. These results indicate that presence of this compound in the hemolymph of A. mylitta provides defense against fungal infection. • An antifungal compound is purified from the hemolymph of tasar silkworm larvae. • Molecular Mass of the purified compound is determined as 531 Da. • Purified compound shows its structure as symmetric bis-decanoic acid derivatives. • Compound disrupts fungal cell wall to exert its antifungal activity. [ABSTRACT FROM AUTHOR]

Summary: Many types of yeast have been studied in the last few years as potential biocontrol agents against different phytopathogenic fungi. Their ability to control plant diseases is mainly through combined modes of action. Among them, antibiosis, competition for nutrients and niches, induction of systemic resistance in plants and mycoparasitism have been the most studied. In previous work, we have established that the epiphytic yeast Pseudozyma aphidis inhibits Botrytis cinerea through induced resistance and antibiosis. Here, we demonstrate that P. aphidis adheres to B. cinerea hyphae and competes with them for nutrients. We further show that the secreted antifungal compounds activate the production of reactive oxygen species and programmed cell death in B. cinerea mycelium. Finally, P. aphidis and its secreted compounds negatively affect B. cinerea hyphae, leading to morphological alterations, including hyphal curliness, vacuolization and branching, which presumably affects the colonization ability and infectivity of B. cinerea. This study demonstrates additional modes of action for P. aphidis and its antifungal compounds against the plant pathogen B. cinerea. [ABSTRACT FROM AUTHOR]

Aims: Microbial biocontrol agents have become an effective option to mitigate the harmfulness of chemical pesticides in recent years. This study demonstrates the control efficacy of Bacillus velezensis CE 100 on the anthracnose causal agent, Colletotrichum gloeosporioides., Methods and Results: In vitro antifungal assays revealed that the culture filtrate and volatile organic compounds of B. velezensis CE 100 strongly restricted the mycelial development of C. gloeosporioides. Moreover, a bioactive compound, butyl succinate, was isolated from the n-butanol crude extract of B. velezensis CE 100 (bce), and identified by liquid chromatography-electrospray ionization hybrid ion-trap and time-of-flight mass spectrometry (LC-ESI-QTOF-MS) and one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR). Treatment with purified butyl succinate at a concentration of 300 μg mL-1 strongly controlled conidial germination of C. gloeosporioides with an inhibition rate of 98.66%, whereas butyl succinate at a concentration of 400 μg mL-1 showed weak antifungal action on the mycelial growth of C. gloeosporioides with an inhibition rate of 31.25%. Scanning electron microscopy revealed that the morphologies of butyl succinate-treated hyphae and conidia of C. gloeosporioides were severely deformed with shriveled and wrinkled surfaces. Furthermore, butyl succinate was able to control carbendazim-resistant C. gloeosporioides, demonstrating that it could be a promising agent for the suppression of other carbendazim-resistant fungal pathogens. An in vivo biocontrol assay demonstrated that the strain ce 100 broth culture and butyl succinate showed higher control efficacy on apple anthracnose than bce., Conclusions: Our findings provide insight into the antifungal potential of B. velezensis  ce 100 and its butyl succinate for efficient control of phytopathogenic fungi, such as C. gloeosporiodes, in plant disease protection. This is the first study to demonstrate the antifungal potential of bacteria-derived butyl succinate for control of C. gloeosporioides., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

An antifungal compound 9, 10-dihydrophenanthrene - 2 - carboxylic acid was isolated from a marine derived bacterium Pseudomonas putida isolated from surface water samples of Karachi fish harbor coast line. The structure was explored using extensive ID- and 2D-NMR spectroscopic techniques. The compound was found to be active against fungal strains obtained from clinical samples whereas strong activity was noted against Candida albicans with a MIC value of 20μg/ml, as the purified compound showed promising anticandidal activity a multidisciplinary approach is needed to explore further this compound as potential pharmacological lead compound against Candida spp and will add in the global hunt for clinically functional antifungal agents. [ABSTRACT FROM AUTHOR]

Paracoccidioidomycosis (PCM) is a systemic mycosis endemic in Latin America, caused by Paracoccidioides spp. A limited number of antifungal agents are available and the search for new compounds has increased. Additionally, nanostructured lipid system (NLS) has emmerged as an interesting strategy to carrier compounds for the treatment of mycosis. In this work, the antifungal efficacy and toxicity of dodecyl gallate (DOD) associated with a NLS was evaluated through in vitro and in vivo tests. DOD showed good in vitro antifungal activity and low toxicity in lung fibroblasts and zebrafish embryos, but no antifungal efficacy in infected mice, which may have been a result of low bioavailability. On the other hand, the association of DOD + NLS was beneficial and resulted in lower toxicity in lung fibroblasts and zebrafish embryos. In addition, NLS + DOD promoted a significant reduction in the fungal burden of mice lungs and could be a potential therapeutic option against PCM. [ABSTRACT FROM AUTHOR]

Rice blast is a devastating fungal disease resulting in major losses to rice crops. Owing to continuous acquisition of resistance by the causal fungus, several fungicide chemicals are no longer effective. Therefore, there is a need to identify natural components and develop new agents to control fungal pathogens. We previously demonstrated that the culture filtrate of Biscogniauxia sp. O821 inhibited infection behavior of Magnaporthe oryzae and subsequent blast lesion formation. In the present study, we isolated a new compound, (3aS,4aR,8aS,9aR)-3a-hydroxy-8a-methyl-3,5-dimethylenedecahydronaphto[2,3-b]furan-2(3H)-one (HDFO), from the culture filtrate of Biscogniauxia sp. O821 and determined its molecular weight as 248. The HDFO structure was determined by electrospray ionization-mass spectrometry and nuclear magnetic resonance spectroscopy after purification with column chromatography and high-performance liquid chromatography. The structure of this antifungal compound was similar to that of alantolactone and isoalantolactone. The growth inhibition zone against M. oryzae in presence of HDFO was observed at Rf 0.5–0.6 on a thin layer chromatography plate. HDFO inhibited conidial germination of M. oryzae in a dose-dependent manner (1–200 ppm). Furthermore, blast lesion formation was significantly suppressed by HDFO at over 5 ppm. These results suggest that HDFO from the culture filtrate of Biscogniauxia sp. O821 can protect rice from rice blast disease caused by M. oryzae. This is the first report that HDFO produced by Biscogniauxia sp. can serve as an antifungal compound against M. oryzae. [ABSTRACT FROM AUTHOR]

Endophytic bacteria have been reported to produce metabolite as antifungal compound. This study was aimed to obtain endophytic bacteria which are able to produce metabolite to control Fusarium sp., a potential seedborne fungi on maize. Endophytic bacteria were screened by hypersensitive test on tobacco leaves and antagonistic test. Endophytic bacteria isolates with high growth inhibitor activity were selected and examined for their metabolite compound. Thre isolates, i.e. Lactobacillus sp. isolate EF14III, Pseudomonas sp. isolate ER1I, dan Aeromonas sp. isolate ER10I has the potential to inhibit Fusarium sp.. Metabolite compound of Pseudomonas sp. isolates ER1I was able to decrease the infection Fusarium sp. by 65.0% in blotter test and decreased infection of Fusarium sp. up to 59.5% and 60.5% in growing on test using water agar and sterile soil, respectively. Cyclohexanone with concentration of 9.68% produced by Pseudomonas sp. isolat ERI1 may play a role as antifungal compound.

An ethyl acetate extract of a culture filtrate (ECF) from an unidentified fungal isolate O821 was evaluated for antifungal activity against the rice pathogen Magnaporthe oryzae. The O821-ECF significantly inhibited spore germination, appressorium formation, and mycelial growth of M. oryzae, and its antifungal activity was heat-stable. It also significantly suppressed the number and size of blast lesions. In an analysis of the ITS sequence of this isolate, it shared similarities with species of the fungus Biscogniauxia. These results suggest that isolate O821 of the genus Biscogniauxia produces a heat-stable antifungal compound(s) in its culture filtrate. [ABSTRACT FROM AUTHOR]

Rhizoctonia solani is the cause of substantial economic loss in many crops. The aim of this study is to investigate biocontrol potential of Bacillus sp. L60 against R. solani and to purify an antifungal compound. In this study, Bacillus sp. L60 demonstrated significant antagonism toward R. solani with the dual culture assay. The antifungal compound was extracted from Bacillus sp. L60 culture supernatant with n-butanol, and identified as N-butyl-tetrahydro-5-oxofuran-2-carboxamide (BT-5O-2C) having molecular weights of 185.1052 Da with the formula C9H15NO3 using NMR and HR-ESI-MS analysis. The minimum inhibitory concentration (MIC) value of the antifungal compound was 256 µg ml−1 against R. solani. Therefore, our results clearly demonstrated BT-5O-2C as well as Bacillus sp. L60 as potential biological control agents for the management of R. solani. [ABSTRACT FROM AUTHOR]

The release of pharmaceuticals into environment has become an increasing concern in recent years. Fungi are part of the microbial flora of many animals, humans and foodstuffs, and some species can cause disease. An antimycotic or antifungal product is one that is used in the treatment of fungal infections. Even at low concentrations, antifungals exert an action against micro-organisms and exhibit selective toxicity towards them. The use of antimicrobials selects for resistant populations of micro-organisms. Development of resistance to antifungals is an increasing problem in veterinary and human medicine.

In this study, we investigated the antifungal activity and mechanism of action of (+)-pinoresinol, a biphenolic compound isolated from the herb Sambucus williamsii,used in traditional medicine. (+)-Pinoresinol displays potent antifungal properties without hemolytic effects on human erythrocytes. To understand the antifungal mechanism of (+)-pinoresinol, we conducted fluorescence experiments on the human pathogen Candida albicans. Fluorescence analysis using 1,6-diphenyl-1,3,5-hexatriene (DPH) indicated that the (+)-pinoresinol caused damage to the fungal plasma membrane. This result was confirmed by using rhodamine-labeled giant unilamellar vesicle (GUV) experiments. Therefore, the present study indicates that (+)-pinoresinol possesses fungicidal activities and therapeutic potential as an antifungal agent for the treatment of fungal infectious diseases in humans.