Broad-spectrum Antifungal Activity Research Articles

Synthesis and antifungal activities of small molecule arylthiazolamine derivatives.

Developing new fungicides to compensate for the deficiencies of existing fungicides resistance in phytopathogenic fungi is a research hotspot in the field of pesticides. Aiming to discover novel template small molecules with excellent antifungal activity, thirty-eight arylthiazolamine derivatives were synthesized through bromination, cyclization, halogenation, and acylation reactions. The synthesized compounds were screened for antifungal activity against ten typical fungal pathogens, and some halogenated arylthiazolamines and amides exhibited excellent broad-spectrum antifungal activity, especially compounds 4m (3.96-47.76μg/mL), 5k (0.10-7.70μg/mL) and 5n (2.08-11.21μg/mL). Among them, compound 5k provided comparable protection and curative effects to chloroticonil and boscalid against B. dothidea and V. mali infection in apple and apple tree branches, respectively, and it could exert antifungal effects by inhibiting the differentiation of mycelium spores, spore germination, and bud tube growth. This study provides high-efficiency and inexpensive candidate compounds for managing of diseases caused by plant pathogenic fungi.

Exploration of antimicrobial activities and mechanisms of biocontrol agent Serratia nematodiphila BC-SKRU-1 against Penicillium digitatum in tangerine fruit

The isolate BC-SKRU-1, identified as Serratia nematodiphila from fungal culture contamination, was investigated for its biocontrol potential against Penicillium digitatum in vitro and on tangerine fruit, with a focus on its mechanisms of action. In vitro tests on PDA plates revealed BC-SKRU-1’s broad-spectrum antifungal activity, achieving up to 74.68% inhibition against seven plant pathogenic fungi, including P. digitatum NKP4321. Bacterial culture filtrates from BC-SKRU-1 (BCF BC-SKRU-1) at 5 to 10% concentrations inhibited NKP4321 mycelial growth in PDB medium (78.41 - 83.68%) more effectively than in PDA medium (46.20 - 72.61%), with complete suppression at 15% (v/v) in both media. BCF BC-SKRU-1's efficacy was comparable to chemical fungicides such as propiconazole®, prochloraz®, and mancozeb®. Notably, BCF BC-SKRU-1 retained its antifungal activity after dilution (1/1000), autoclaving (at 121°C), and storage (at -20°C). In vivo evaluations on tangerine fruit showed significant reduction in the severity of postharvest green mold disease with BC-SKRU-1 treatments, especially at a concentration of 108 CFU mL-1. Both preventive and curative applications were effective, with curative treatments being more successful. Mechanistic studies indicated that BCF BC-SKRU-1 reduces intracellular ergosterol content, compromises plasma membrane integrity, and attenuates antioxidant defense activities (SOD, CAT, GSH, GSSG, and GSH/GSSG ratio). These findings highlight BC-SKRU-1 and its metabolites as promising biocontrol agents against green mold in tangerine fruits and provide insights into their antifungal mechanisms.

Discovery of Highly Potent Triazole Derivatives with Broad-Spectrum Antifungal Activity Based on Iodiconazole

The widespread use of broad-spectrum antibiotics, the growing number of immunocompromised individuals, and the emergence of drug-resistant strains have resulted in the increasing incidence and mortality of invasive fungal infections. Azole drugs are the primary treatment for invasive fungal infections, and Iodiconazole is a potent azole drug with strong antifungal activity, but its stability is poor. In order to improve stability, a series of triazole compounds containing ethynyl group were designed and synthesized. Most of the compounds showed strong inhibitory activity against pathogenic fungi, among which compound 20l showed excellent inhibitory activity against pathogenic fungi and drug-resistant fungi. Importantly, and the stability of 20l (T1/2 = 30.2 min) was obviously improved compared with Iodiconazole (T1/2 = 4.39 min). In addition, the preferred compound 20l can prevent fungal phase transition and the formation of fungal biofilm, and show satisfactory fungicidal activity. In addition, the compound 20l was almost non-toxic to mammalian HUVEC cell and 293T cell. In vivo pharmacokinetic studies showed that 20l had acceptable pharmacokinetic properties. These results strongly demonstrate that compound 20l was worth further investigation as a potential antifungal inhibitor.

Design, Synthesis, and Antifungal Evaluation of Novel Pyrazole-5-sulfonamide Derivatives for Plant Protection.

To develop further novel environmentally friendly antifungal agents with high efficacy, a series of pyrazole-5-sulfonamide derivatives were designed and synthesized by using the active molecules synthesized in previous works as lead compounds. Their antifungal activities were evaluated in vitro against ten highly destructive plant pathogenic fungi. The bioassay results indicated that more than half of the target compounds displayed potent antifungal activities (inhibition rate ≥85%) against Valsa mali and Sclerotinia sclerotiorum at 20 mg/L. Among them, compound C22 exhibited significant broad-spectrum antifungal activities against V. mali, S. sclerotiorum, Rhizoctonia solani, Botrytis cinerea, and Trichoderma viride, with EC50 values of 0.45, 0.49, 3.06, 0.57, and 1.43 mg/L, respectively. Moreover, compounds C21 and C22 exhibited remarkable protective effects on apple Valsa canker similar to tebuconazole (89.5%) at 50 mg/L. Preliminary antifungal mechanism investigations demonstrated that compound C22 may have inhibited V. mali mycelial growth by inducing oxidative damage to the mycelium and compromising the integrity of the cell membrane. Meanwhile, compounds C21 and C22 exhibited no obvious toxicity to worker bees (Apis mellifera ligustica). Taken together, these pyrazole-5-sulfonamide derivatives, particularly compound C22, possess huge potential to be developed as novel environmentally friendly fungicides with high efficacy.

Recent Advances in Phenazine Natural Products: Biosynthesis and Metabolic Engineering.

Phenazine natural products are a class of nitrogen-containing heterocyclic compounds produced by microorganisms. The tricyclic ring molecules show various chemical structures and extensive pharmacological activities, such as antimicrobial, anticancer, antiparasitic, anti-inflammatory, and insecticidal activities, with low toxicity to the environment. Since phenazine-1-carboxylic acid has been developed as a registered biopesticide, the application of phenazine natural products will be promising in the field of agriculture pathogenic fungi control based on broad-spectrum antifungal activity, minimal toxicity to the environment, and improvement of crop production. Currently, there are still plenty of intriguing hidden biosynthetic pathways of phenazine natural products to be discovered, and the titer of naturally occurring phenazine natural products is insufficient for agricultural applications. In this review, we spotlight the progress regarding biosynthesis and metabolic engineering research of phenazine natural products in the past decade. The review provides useful insights concerning phenazine natural products production and more clues on new phenazine derivatives biosynthesis.

Design and synthesis of spiro[pyrrolidine-3,3′-quinoline]-2,2′-dione derivatives as novel antifungal agents targeting chitin synthase

A novel spiro [pyrrolidine-3′,3′-quinoline]-2,2′-dione scaffold was constructed using fragments of quinoline and pyrrolidine. Subsequently, two series of derivatives were designed based on this scaffold. The enzyme inhibition experiments revealed that all designed compounds had moderate to good inhibitory activity against chitin synthase (CHS). The inhibitory effects of compounds 5i, 5j, 8i and 8n were approximately equal to that of control drug polyoxin B (PB, IP = 86.4 ± 2.9 %, IC50 = 0.082 ± 0.013 mM) which is a well-established CHS inhibitor. The results from enzyme kinetic parameters assays proved that these compounds act as non-competitive inhibitors of CHS. The sorbitol protection experiments demonstrated the tested compounds disrupted the synthesis of cell wall, which further verified that the target of these compounds is CHS. The experiments of antimicrobial showed that compounds 5b, 5f, 5i, 5j, 8f, 8i, 8m, 8n and 8o had strong antifungal activity against the four tested pathogenic fungi strains frequently emerging in clinical setting, with MIC values ranging from 4 to 32 μg/mL, which were either superior to or comparable with those of PB or fluconazole. Furthermore, these compounds displayed synergistic or additive effects when combined with fluconazole and these active compounds also showed promising activity against fluconazole-resistant and micafungin-resistant fungi variants. The result of antimicrobial experiments indicated that these compounds had minimal activity to tested bacterial strains. This suggests that they had selective antifungal activity. The results of ADME prediction, in conjunction with the cytotoxicity assay results, indicated that these compounds had favorable pharmacokinetic profiles and low toxicity. In addition, molecular docking studies illustrated that the compound had a strong affinity with the CHS, which was consistent with the results of enzymatic assays. These findings indicated that the designed compounds are non-competitive inhibitors of CHS with good selectivity and broad-spectrum antifungal activity, and possess significant antifungal activity against drug-resistant fungi, suggesting their potential as lead compounds for the development of novel drugs against drug-resistant fungi.

Design, Bioactivity, and Action Mechanism of Pyridinecarbaldehyde Phenylhydrazone Derivatives with Broad-Spectrum Antifungal Activity.

Replacing old pesticides with new pesticide varieties has been the main means to solve pesticide resistance. Therefore, it is necessary to research and develop new antifungal agents for plant protection. In this study, a series of pyridinecarbaldehyde phenylhydrazone derivatives were designed and evaluated for their inhibition activity on plant pathogenic fungi to search for novel fungicide candidates. Picolinaldehyde phenylhydrazone (1) and nicotinaldehyde phenylhydrazone (2) were identified as promising antifungal lead scaffolds. The 4-fluorophenylhydrazone derivatives (1a and 2a) of 1 and 2 showed highly effective and broad-spectrum inhibition activity in vitro on 11 phytopathogenic fungi with EC50 values of 0.870-3.26 μg/mL, superior to the positive control carbendazim in most cases. The presence of the 4-fluorine atom on the phenyl showed a remarkable activity enhancement effect. Compound 1a at 300 μg/mL provided almost complete protection against infection of Alternaria solani on tomatoes over the post-treatment 9 days and high safety to germination of plant seeds. Furthermore, 1a showed strong inhibition activity with an IC50 value of 0.506 μg/mL on succinate dehydrogenase in A. solani. Molecular docking showed that both 1a and 2a can well bind to the ubiquinone-binding region of SDH by the conventional hydrogen bond, carbon-hydrogen bond, π-π or π-amide interaction, π-alkyl interaction, X---F (X = N, C, or H) interaction, and van der Waal forces. Meanwhile, scanning and transmission electron analysis displayed that 1a destroyed the morphology of mycelium and the structure of the cell membrane of A. solani. Fluorescent staining analysis revealed that 1a changed the mitochondrial membrane potential and cell membrane permeability. Thus, pyridinecarbaldehyde phenylhydrazone compounds emerged as novel antifungal lead scaffolds, and 1a and 2a can be considered promising candidates for the development of new agricultural fungicides.

Innovative Ag@Cu/white sand and polysaccharide based nanocomposites: A simple route to conductive and antibacterial paper coatings

In this work, we presented a simple method for preparing nanocomposite pigments via doping the naturally occurring white sand pigment using silver (Ag) and copper (Cu). The nanocomposites (Ag@WS, Cu@WS, and Ag&Cu@WS) were produced in two suspensions at 25% and 50% w/v concentrations, utilizing casein (Cas) and nanostarch (NanoSt) as bio-friendly binders. The prepared nanocomposite pigments were characterized using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction crystallography (XRD), and dynamic light scattering (DLS), as well as topographical analysis including scanning electron microscopy (SEM), EDX, and transmission electron microscopy (TEM). The coated paper sheets were characterized using topographical analysis and mechanical, optical, physical, and electrical properties. The antimicrobial activity assay was carried out on the coated paper sheets using the most infectious microbial pathogens. FTIR and XRD analyses confirmed the presence of Cas and NanoSt in the samples. The addition of white sand (WS) and doped nano-metals influenced the crystallographic structure of the polysaccharides. A small amount of Ag, Cu, and Ag&Cu nanoparticles was investigated through EDX. The white sand was a plate structure decorated with spherical Ag, Cu, and Ag&Cu nanoparticles, with an average size of about 147, 105, and 78 nm, as observed from TEM images and DLS. The antimicrobial activity assays revealed that nanocomposite pigment embedded with bimetallic Ag&Cu nanoparticles exhibited a synergistic effect, resulting in enhanced broad-spectrum antibacterial and antifungal activities. The electrical performance of the coated paper was enhanced by using Ag&Cu@WS nanocomposite pigment. Incorporating 50% Cu@WS into the coating resulted in significant improvements in the mechanical properties of the paper. Specifically, tensile strength increased by 6.3 kN/m, tensile index by 69.06 N·m/g, stretching by 1.68%, tensile energy absorption by 71.4 J/m2, burst strength by 257.75 kPa, and burst index by 2.83 kPa·m2/g. Furthermore, incorporating 50% Ag&Cu@WS increased the opacity to 93.2% and reduced the roughness of the coated paper by 36% compared to using 50% Ag@WS or 50% Cu@WS alone.

Analogues of the anti-malaria drug mefloquine have broad spectrum antifungal activity and are efficacious in a model of disseminated Candida auris infection.

Only three classes of antifungal drugs are currently in clinical use. Here, we report that derivatives of the malarial drug mefloquine have broad spectrum antifungal activity including difficult to treat molds and endemic fungi. Pharmacokinetic and efficacy studies of NSC-4377 indicate it penetrates the central nervous system and is active against Candida auris in vivo. These data strongly support the further development of mefloquine analogs as a potentially new class of antifungal molecules.

Isolation, Identification, and Biocontrol Mechanisms of Endophytic Burkholderia arboris DHR18 from Rubber Tree against Red Root Rot Disease.

Red root rot disease is a devastating fungal disease of rubber trees caused by Ganoderma pseudoferreum (Wakef). Biocontrols using beneficial microorganisms are safe and sustainable. We isolated a DHR18 endophytic bacterium from a healthy rubber tree to obtain a new efficient antagonistic bacterium for red root rot disease affecting rubber trees and evaluated the mechanism of action involved using a double culture assay, genome annotation analysis, and the ethyl acetate extraction method. The results revealed that the DHR18 strain inhibits G. pseudoferreum growth and has broad-spectrum antifungal activity by secreting cell wall hydrolases (proteases and chitinases), indole-3-acetic acid, and siderophores. Furthermore, it fixes nitrogen and is involved in biofilm formation and phosphate solubilisation, improving disease resistance and tree growth. The results showed that the antifungal substances secreted by DHR18 are mainly lipopeptides. Simultaneously, DHR18 enhanced the rubber tree resistance to G. pseudoferreum by increasing the activities of defence enzymes superoxide dismutase, phenylalanine ammonia lyase, peroxidase, catalase, and polyphenol oxidase. The results indicate that B. arboris DHR18 has biocontrol potential and could be used as a candidate strain for the control of red root rot disease in rubber trees.

Anti-Aflatoxigenic Burkholderia contaminans BC11-1 Exhibits Mycotoxin Detoxification, Phosphate Solubilization, and Cytokinin Production.

The productivity and quality of agricultural crops worldwide are adversely affected by disease outbreaks and inadequate nutrient availability. Of particular concern is the potential increase in mycotoxin prevalence due to crop diseases, which poses a threat to food security. Microorganisms with multiple functions have been favored in sustainable agriculture to address such challenges. Aspergillus flavus is a prevalent aflatoxin B1 (AFB1)-producing fungus in China. Therefore, we wanted to obtain an anti-aflatoxigenic bacterium with potent mycotoxin detoxification ability and other beneficial properties. In the present study, we have isolated an anti-aflatoxigenic strain, BC11-1, of Burkholderia contaminans, from a forest rhizosphere soil sample obtained in Luzhou, Sichuan Province, China. We found that it possesses several beneficial properties, as follows: (1) a broad spectrum of antifungal activity but compatibility with Trichoderma species, which are themselves used as biocontrol agents, making it possible to use in a biocontrol mixture or individually with other biocontrol agents in an integrated management approach; (2) an exhibited mycotoxin detoxification capacity with a degradation ratio of 90% for aflatoxin B1 and 78% for zearalenone, suggesting its potential for remedial application; and (3) a high ability to solubilize phosphorus and produce cytokinin production, highlighting its potential as a biofertilizer. Overall, the diverse properties of BC11-1 render it a beneficial bacterium with excellent potential for use in plant disease protection and mycotoxin prevention and as a biofertilizer. Lastly, a pan-genomic analysis suggests that BC11-1 may possess other undiscovered biological properties, prompting further exploration of the properties of this unique strain of B. contaminans. These findings highlight the potential of using the anti-aflatoxigenic strain BC11-1 to enhance disease protection and improve soil fertility, thus contributing to food security. Given its multiple beneficial properties, BC11-1 represents a valuable microbial resource as a biocontrol agent and biofertilizer.

Inhibitory mechanism of 4-ethyl-1,2-dimethoxybenzene produced by Streptomyces albidoflavus strain ML27 against Colletotrichum gloeosporioides

Actinomycetes have emerged as significant biocontrol resources due to their rich array of bioactive natural products. While much research has historically focused on secondary metabolites isolated from their fermentation broth, there remains a dearth of reports on their volatile organic compounds (VOCs). Here, strain ML27, isolated from soil, was identified as Streptomyces albidoflavus based on morphological features, physiological, biochemical, and molecular characteristics (16S rRNA, atpD, recA, and rpoB gene sequences). VOCs from S. albidoflavus strain ML27 were effectively captured using solid-phase microextraction (SPME) and tentatively identified through gas chromatography–mass spectrometry (GC/MS). Among these compounds, 4-ethyl-1,2-dimethoxybenzene exhibited broad-spectrum antifungal activity and demonstrated efficacy in controlling citrus anthracnose, with a control efficacy of 86.67%. Furthermore, the inhibitory mechanism of 4-ethyl-1,2-dimethoxybenzene against Colletotrichum gloeosporioides was revealed. Results indicated that 4-ethyl-1,2-dimethoxybenzene induced swelling, deformity, and breakage in C. gloeosporioides mycelia, and significantly inhibited spore germination. Transcriptome analysis revealed that 4-ethyl-1,2-dimethoxybenzene inhibited the growth and development of C. gloeosporioides primarily by disrupting energy metabolism and the integrity of the cell wall and membrane. Based on these results, it is promising to develop 4-ethyl-1,2-dimethoxybenzene as a novel biopesticide for controlling citrus anthracnose.

Synthesis and Biological Activities of C1-Substituted Acylhydrazone β-Carboline Analogues as Antifungal Candidates.

In our ongoing work to create potential antifungal agents, we synthesized and tested a group of C1-substituted acylhydrazone β-carboline analogues 9a-o and 10a-o for their effectiveness against Valsa mali, Fusarium solani, Fusarium oxysporum, and Fusarium graminearum. Their compositions were analyzed using different spectral techniques, such as 1H/13C NMR and HRMS, with the structure of 9l being additionally confirmed through X-ray diffraction. The antifungal evaluation showed that, among all the target β-carboline analogues, compounds 9n and 9o exhibited more promising and broad-spectrum antifungal activity than the commercial pesticide hymexazol. Several intriguing findings regarding structure-activity relationships (SARs) were examined. In addition, the cytotoxicity test showed that these acylhydrazone β-carboline analogues with C1 substitutions exhibit a preference for fungi, with minimal harm to healthy cells (LO2). The reported findings provide insights into the development of β-carboline analogues as new potential antifungal agents.

Microscopic and Transcriptomic Analyses to Elucidate Antifungal Mechanisms of Bacillus velezensis TCS001 Lipopeptides against Botrytis cinerea.

Botrytis cinerea is an important fungal pathogen that causes gray mold disease in plants. Previously, Bacillus velezensis TCS001 live culture presented broad-spectrum antifungal activity against various plant pathogenic fungi and oomycetes, particularly B. cinerea. Here, the bioactivity of lipopeptides produced by TCS001 against B. cinerea was investigated. The IC50 values of the crude lipopeptide extract (CLE) from TCS001 to suppress mycelial growth and conidial germination were 14.20 and 49.39 mg/L, respectively. SEM and TEM imaging revealed that CLE caused morphological deformities and ultrastructural changes in the mycelium. Transcriptomic analyses combined with ΔBcpsd mutant construction demonstrated that the CLE could confer antifungal activity via suppressing Bcpsd expression in the pathogen. In addition, the CLE activated the plant immune system by increasing the content of defense-related enzymes and the expression of marker genes in immunity signaling pathways in cucumber plants. Therefore, TCS001 CLE could be potentially developed into biopesticides for the biocontrol of gray mold disease.

Inhibitor of cardiolipin biosynthesis-related enzyme MoGep4 confers broad-spectrum anti-fungal activity.

Plant pathogens cause devastating diseases, leading to serious losses to agriculture. Mechanistic understanding of pathogenesis of plant pathogens lays the foundation for the development of fungicides for disease control. Mitophagy, a specific form of autophagy, is important for fungal virulence. The role of cardiolipin, mitochondrial signature phospholipid, in mitophagy and pathogenesis is largely unknown in plant pathogenic fungi. The functions of enzymes involved in cardiolipin biosynthesis and relevant inhibitors were assessed using a set of assays, including genetic deletion, plant infection, lipidomics, chemical-protein interaction, chemical inhibition, and field trials. Our results showed that the cardiolipin biosynthesis-related gene MoGEP4 of the rice blast fungus Magnaporthe oryzae regulates growth, conidiation, cardiolipin biosynthesis, and virulence. Mechanistically, MoGep4 regulated mitophagy and Mps1-MAPK phosphorylation, which are required for virulence. Chemical alexidine dihydrochloride (AXD) inhibited the enzyme activity of MoGep4, cardiolipin biosynthesis and mitophagy. Importantly, AXD efficiently inhibited the growth of 10 plant pathogens and controlled rice blast and Fusarium head blight in the field. Our study demonstrated that MoGep4 regulates mitophagy, Mps1 phosphorylation and pathogenesis in M. oryzae. In addition, we found that the MoGep4 inhibitor, AXD, displays broad-spectrum antifungal activity and is a promising candidate for fungicide development.

Antifungal properties of Andrographis paniculata extracts as potential biofungicides: a review

Andrographis paniculata, commonly known as the king of bitters, has been studied for its potential as a natural fungicide due to its several bioactive compounds, including andrographolides. The primary objective of this study was to demonstrate the possible efficacy of A. paniculata against a wide range of fungal diseases in terms of the properties of A. paniculata extracts that could potentially be used as a bio-fungicide. The presence of alkaloids, flavonoids, phenols, terpene derivatives compounds, and other phytochemical constituents makes the A. paniculata extract more precise in its mode of action against the variety of phytopathogenic fungi. Various physiological and biochemical changes in crops result in unintended physiological issues, degradation, and subsequent financial losses during storage. Synthetic fungicides have been applied to control plant diseases caused by phytopathogenic fungi. However, their application has drawn criticism because of adverse effects. Therefore, a possible solution would be to apply botanical fungicides, which have been proven to be generally non-phytotoxic, systemic, and rapidly disposable in nature. Botanical fungicides are endowed with benefits such as resistance inhibition, being ecofriendly, effective, elective, and more affordable than synthetic fungicides. Therefore, the medicinal herb, A. paniculata has been proven to be effective in controlling pathogens of agricultural crops and increased their shelf-life when used as a substitute for synthetic chemicals. Due to their antimicrobial capabilities against fungal, bacterial, and viral infections, various medicinal herbs have been investigated and employed for crude extraction globally. Andrographis paniculata extracts have broad-spectrum antifungal activity, indicating potential efficacy against a diverse variety of fungal infections.

A series of pyrimidine-based antifungals with anti-mold activity disrupt ER function in Aspergillus fumigatus.

Fungal infections are a major contributor to morbidity and mortality among immunocompromised populations. Moreover, fungal disease caused by molds are difficult to treat and are associated with particularly high mortality. To address the need for new mold-active antifungal drugs, we performed a high-throughput screen with Aspergillus fumigatus, the most common pathogenic mold. We identified a novel, pyrimidine-based chemical scaffold with broad-spectrum antifungal activity including activity against several difficult-to-treat molds. A chemical genetics screen of Saccharomyces cerevisiae suggested that this compound may target the endoplasmic reticulum (ER) and perturb ER function and/or homeostasis. Consistent with this model, this compound induces the unfolded protein response and inhibits secretion of A. fumigatus collagenases. Initial cytotoxicity and pharmacokinetic studies show favorable features including limited mammalian cell toxicity and bioavailability in vivo. Together, these data support the further medicinal chemistry and pre-clinical development of this pyrimidine scaffold toward more effective treatments for life-threatening invasive mold infections.IMPORTANCEInvasive fungal diseases are life-threatening infections caused by fungi in immunocompromised individuals. Currently, there are only three major classes of antifungal drugs available to treat fungal infections; however, these options are becoming even more limited with the global emergence of antifungal drug resistance. To address the need for new antifungal therapies, we performed a screen of chemical compounds and identified a novel molecule with antifungal activity. Initial characterization of this compound shows drug-like features and broad-spectrum activity against medically important fungi. Together, our results support the continued development of this compound as a potential future therapy for these devastating fungal infections.

Berberine Degradation Characteristics and its Degradation Pathway by a Newly Isolated Berberine-Utilizing Agrobacterium.

Berberine (BBR) is widely used as a botanical pesticide due to its broad-spectrum antibacterial and antifungal activities. However, BBR degradation pathway in soil microorganisms, which determines its impact on soil environment, remains poorly understood. Herein, a novel BBR-degrading bacterium Agrobacterium sp. V1 was isolated and characterized. Agrobacterium sp. V1 was able to utilize BBR as the sole carbon source for cell growth, and 50μg/mL of BBR was completely degraded within 48h. To reveal the possible BBR degradation pathway, whole genome sequencing of Agrobacterium sp. V1 was conducted, and proteins in Agrobacterium sp. V1 were aligned with enzymes involved in BBR biosynthesis in Rhizoma Coptidis. The results indicated that more than 60% of enzymes in BBR biosynthesis pathway had orthologs in Agrobacterium sp. V1. Combined with the primary mass spectra of BBR metabolites, a novel BBR degradation pathway in this bacterium was proposed. In summary, the proposed BBR degradation pathway offered new insights into the impact of BBR to the environment and also provided a reference for studying BBR metabolism in microorganisms.

Antifungal Activity and Possible Mechanism of Streptomyces nojiriensis 9-13 Against Mycogone sp., Causing Wet Bubble Disease on Agaricus bisporus.

Wet bubble disease (WBD) in Agaricus bisporus caused by Mycogone species imposes a substantial economic loss to mushroom production in China. Currently, fungicide application is the main method to control WBD. However, excessive use of fungicides is challenged by the appearance of resistance and food safety. Therefore, it is necessary to explore safe and efficient strategies to control WBD. Strain 9-13, isolated from the rhizosphere soil of Taxus chinensis, showed strong inhibitory activity against three Mycogone species. According to morphological and biochemical characteristics and multilocus phylogenetic analysis, the strain was identified as Streptomyces nojiriensis. In addition, strain 9-13 extracts significantly inhibited mycelial growth and spore germination of M. perniciosa, M. rosea, and M. xinjiangensis in vitro. Strain 9-13 and its extracts also exhibited broad-spectrum antifungal activities against 12 selected plant pathogenic fungi. Scanning electron microscopic observations showed that the extracts destroyed mycelial structure, inducing mycelia to twist and shrink. Moreover, transmission electron microscopy revealed that the extracts resulted in severe plasmolysis, rupture of the cell membrane, and a decrease in cell inclusions, and the cell wall had a rough and uneven surface. Notably, the extracts obviously reduced disease severity and incidence of WBD by from 83.85 to 87.32% in fruiting bodies and 77.36% in mushroom beds and maintained fruiting time and color on harvested mushrooms. Collectively, these results clearly indicate that S. nojiriensis 9-13 is a promising biocontrol agent to control WBD on A. bisporus.

MALDI-TOF MS profiling and antifungal activity of lactic acid bacteria from kunu aya, a tiger nut traditional beverage of Nigeria

The application of matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) as a technique for high-throughput characterization of microbiota in African traditional foods is limited. Kunu aya is an unexplored reservoir of protective lactic acid bacteria (LAB) strains. A total of 26 LAB isolates from kunu aya were identified using MALDI-TOF MS as five species, including Lactiplantibacillus plantarum (14), Pediococcus pentosaceus (9), Levilactobacillus brevis (1), Lactiplantibacillus pentosus (1), and Weissella paramesenteroides (1). Antifungal activity screening of all LAB strains by agar-plate overlay assay revealed inhibition against Penicillium crustosum FOO5BA and/or Aspergillus flavus FOO8BA by six strains: L. plantarum (3), Pd. pentosaceus (1), L. brevis (1), and W. paramesenteroides (1). The six LAB strains demonstrated broad spectrum antifungal activity, when tested against eight molds from plant-based foods, inhibiting 4 to 7 molds, with average inhibition scores (AIS) of 0.75–2.75. L. brevis MKAIA1b showed the strongest antifungal activity (AIS; 2.75). Agar diffusion method revealed that antifungal activity against all target molds was retained in cell-free supernatants of L. brevis MKAIA1b, with the highest inhibition demonstrated against P. citrinum 3AS1 (15.34%). In challenged kunu aya fermented with L. brevis MKAIA1b or L. brevis MKAIA1b and L. plantarum TMP051, the growth of A. flavus F008BA, Fusarium langesthiae 13,014, and P. crustosum FOO5BA was delayed beyond 7 days at ambient temperature (25 °C). In this study, MALDI-TOF MS was an effective tool to reveal the species composition of LAB in kunu aya, which harbors LAB strains that displayed strong antifungal activity, while showing great potentials as a protective culture and for bio-control.