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1.
Sci Rep ; 14(1): 15601, 2024 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-38971913

RESUMO

The electrical potential of the mycelia of a cord-forming wood decay fungus, Pholiota brunnescens, was monitored for over 100 days on a plain agar plate during the colonization onto a wood bait. Causality analyses of the electrical potential at different locations of the mycelium revealed a clear and stable causal relationship with the directional flow of the electrical potential from the hyphae at the bait location to other parts of the mycelium. However, this causality disappeared after 60 days of incubation, coinciding with the onset of slow electrical oscillation at the bait location, which occurred over one week per oscillation cycle. We speculated that the hyphae that initially colonized the bait may act as a temporary activity center, which generates electrical signals to other parts of the mycelium, thereby facilitating the colonization of the entire mycelial body to the bait. The week-long electrical oscillation represents the longest oscillation period ever recorded in fungi and warrants further investigation to elucidate its function and stability in response to environmental stimuli.


Assuntos
Micélio , Micélio/fisiologia , Hifas/fisiologia , Ascomicetos/fisiologia , Madeira/microbiologia
2.
BMC Microbiol ; 24(1): 255, 2024 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-38982358

RESUMO

BACKGROUND: Alternaria alternata is the primary pathogen of potato leaf spot disease, resulting in significant potato yield losses globally. Endophytic microorganism-based biological control, especially using microorganisms from host plants, has emerged as a promising and eco-friendly approach for managing plant diseases. Therefore, this study aimed to isolate, identify and characterize the endophytic fungi from healthy potato leaves which had great antifungal activity to the potato leaf spot pathogen of A. alternata in vitro and in vivo. RESULTS: An endophytic fungal strain SD1-4 was isolated from healthy potato leaves and was identified as Talaromyces muroii through morphological and sequencing analysis. The strain SD1-4 exhibited potent antifungal activity against the potato leaf spot pathogen A. alternata Lill, with a hyphal inhibition rate of 69.19%. Microscopic and scanning electron microscope observations revealed that the strain SD1-4 grew parallel to, coiled around, shrunk and deformed the mycelia of A. alternata Lill. Additionally, the enzyme activities of chitinase and ß-1, 3-glucanase significantly increased in the hyphae of A. alternata Lill when co-cultured with the strain SD1-4, indicating severe impairment of the cell wall function of A. alternata Lill. Furthermore, the mycelial growth and conidial germination of A. alternata Lill were significantly suppressed by the aseptic filtrate of the strain SD1-4, with inhibition rates of 79.00% and 80.67%, respectively. Decrease of leaf spot disease index from 78.36 to 37.03 was also observed in potato plants treated with the strain SD1-4, along with the significantly increased plant growth characters including plant height, root length, fresh weight, dry weight, chlorophyll content and photosynthetic rate of potato seedlings. CONCLUSION: The endophyte fungus of T. muroii SD1-4 isolated from healthy potato leaves in the present study showed high biocontrol potential against potato leaf spot disease caused by A. alternata via direct parasitism or antifungal metabolites, and had positive roles in promoting potato plant growth.


Assuntos
Alternaria , Endófitos , Doenças das Plantas , Folhas de Planta , Solanum tuberosum , Talaromyces , Alternaria/crescimento & desenvolvimento , Alternaria/fisiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Solanum tuberosum/microbiologia , Talaromyces/genética , Talaromyces/crescimento & desenvolvimento , Endófitos/fisiologia , Endófitos/isolamento & purificação , Endófitos/genética , Folhas de Planta/microbiologia , Hifas/crescimento & desenvolvimento , Antibiose , Quitinases/metabolismo , Agentes de Controle Biológico , Controle Biológico de Vetores/métodos
3.
Phytopathology ; 114(7): 1502-1514, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-39023506

RESUMO

Late blight, caused by the notorious pathogen Phytophthora infestans, poses a significant threat to potato (Solanum tuberosum) crops worldwide, impacting their quality as well as yield. Here, we aimed to investigate the potential use of cinnamaldehyde, carvacrol, and eugenol as control agents against P. infestans and to elucidate their underlying mechanisms of action. To determine the pathogen-inhibiting concentrations of these three plant essential oils (PEOs), a comprehensive evaluation of their effects using gradient dilution, mycelial growth rate, and spore germination methods was carried out. Cinnamaldehyde, carvacrol, and eugenol were capable of significantly inhibiting P. infestans by hindering its mycelial radial growth, zoospore release, and sporangium germination; the median effective inhibitory concentration of the three PEOs was 23.87, 8.66, and 89.65 µl/liter, respectively. Scanning electron microscopy revealed that PEOs caused the irreversible deformation of P. infestans, resulting in hyphal shrinkage, distortion, and breakage. Moreover, propidium iodide staining and extracellular conductivity measurements demonstrated that all three PEOs significantly impaired the integrity and permeability of the pathogen's cell membrane in a time- and dose-dependent manner. In vivo experiments confirmed the dose-dependent efficacy of PEOs in reducing the lesion diameter of potato late blight. Altogether, these findings provide valuable insight into the antifungal mechanisms of PEOs vis-à-vis late blight-causing P. infestans. By utilizing the inherent capabilities of these natural compounds, we could effectively limit the harmful impacts of late blight on potato crops, thereby enhancing agricultural practices and ensuring the resilience of global potato food production.


Assuntos
Cimenos , Eugenol , Óleos Voláteis , Phytophthora infestans , Doenças das Plantas , Solanum tuberosum , Phytophthora infestans/efeitos dos fármacos , Phytophthora infestans/fisiologia , Solanum tuberosum/microbiologia , Óleos Voláteis/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Eugenol/farmacologia , Cimenos/farmacologia , Monoterpenos/farmacologia , Micélio/efeitos dos fármacos , Micélio/crescimento & desenvolvimento , Óleos de Plantas/farmacologia , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Esporos/efeitos dos fármacos , Esporos/fisiologia , Acroleína/análogos & derivados
4.
Cell Host Microbe ; 32(6): 779-781, 2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38870893

RESUMO

In a recent issue of Nature, Zhao et al. have demonstrated that Streptomyces spp. produce "umbrella"-shaped polymorphic toxin particles, a novel class of non-lethal toxins that gently inhibit competitors by arresting hyphal growth in closely related bacteria, unveiling a unique bacterial defense strategy in microbial ecological interactions.1.


Assuntos
Toxinas Bacterianas , Streptomyces , Streptomyces/metabolismo , Toxinas Bacterianas/metabolismo , Toxinas Bacterianas/toxicidade , Antibiose , Hifas/crescimento & desenvolvimento , Interações Microbianas
5.
Fungal Biol ; 128(4): 1868-1875, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38876539

RESUMO

In the development of fungal based materials for applications in construction through to biomedical materials and fashion, understanding how to regulate and direct growth is key for gaining control over the form of material generated. Here, we show how simple 'chemical food' cues can be used to manipulate the growth of fungal networks by taking Aspergillus niger as an exemplar species. Chemotrophic responses towards a range of nitrogen and carbon containing biomolecules including amino acids, sugars and sugar alcohols were quantified in terms of chemotrophic index (CI) under a range of basal media compositions (low and high concentrations of N and C sources). Growth of filamentous networks was followed using fluorescence microscopy at single time points and during growth by an AI analytical approach to explore chemo sensing behaviour of the fungus when exposed to pairs (C-C, C-N, N-N) of biomolecules simultaneously. Data suggests that the directive growth of A. niger can be controlled towards simple biomolecules with CI values giving a good approximation for expected growth under a range of growth conditions. This is a first step towards identifying conditions for researcher-led directed growth of hyphae to make mycelial mats with tuneable morphological, physicochemical, and mechanical characteristics.


Assuntos
Aspergillus niger , Meios de Cultura , Hifas , Nitrogênio , Aspergillus niger/crescimento & desenvolvimento , Aspergillus niger/metabolismo , Hifas/crescimento & desenvolvimento , Meios de Cultura/química , Nitrogênio/metabolismo , Carbono/metabolismo , Aminoácidos/metabolismo , Microscopia de Fluorescência
6.
Sci Rep ; 14(1): 13797, 2024 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-38877139

RESUMO

In filamentous fungi, microtubules are important for polar growth and morphological maintenance and serve as rails for intracellular trafficking. The molecular mechanisms associated with microtubules have been analyzed. However, little is known about when and where tubulin, a component of microtubules, is biosynthesized in multinuclear and multicellular filamentous fungi. In this study, we visualized microtubules based on the enhanced green fluorescence protein (EGFP)-labeled α-tubulin and ß-tubulin mRNA tagged by the EGFP-mediated MS2 system in living yellow Koji mold Aspergillus oryzae cells in order to understand the spatiotemporal production mechanism of tubulin. We found that mRNA of btuA, encoding for ß-tubulin, localized at dot-like structures through the apical, middle and basal regions of the hyphal cells. In addition, some btuA mRNA dots showed microtubule-dependent motor protein-like dynamics in the cells. Furthermore, it was found that btuA mRNA dots were decreased in the cytoplasm just before mitosis but increased immediately after mitosis, followed by a gradual decrease. In summary, the localization and abundance of ß-tubulin mRNA is spatiotemporally regulated in living A. oryzae hyphal cells.


Assuntos
Aspergillus oryzae , Microtúbulos , RNA Mensageiro , Tubulina (Proteína) , Aspergillus oryzae/genética , Aspergillus oryzae/metabolismo , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Microtúbulos/metabolismo , Hifas/genética , Hifas/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Regulação Fúngica da Expressão Gênica , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo
7.
Microbes Environ ; 39(2)2024.
Artigo em Inglês | MEDLINE | ID: mdl-38866480

RESUMO

Mn(II)-oxidizing microorganisms are considered to play significant roles in the natural geochemical cycles of Mn and other heavy metals because the insoluble biogenic Mn oxides (BMOs) that are produced by these microorganisms adsorb other dissolved heavy metals and immobilize them as precipitates. In the present study, a new Mn(II)-oxidizing fungal strain belonging to the ascomycete genus Periconia, a well-studied plant-associating fungal genus with Mn(II)-oxidizing activity that has not yet been exami-ned in detail, was isolated from natural groundwater outflow sediment. This isolate, named strain TS-2, was confirmed to oxidize dissolved Mn(II) and produce insoluble BMOs that formed characteristic, separately-located nodules on their hyphae while leaving major areas of the hyphae free from encrustation. These BMO nodules also adsorbed and immobilized dissolved Cu(II), a model analyte of heavy metals, as evidenced by elemental mapping ana-lyses of fungal hyphae-BMO assemblages using a scanning electron microscope with energy-dispersive X-ray spectroscopy (SEM-EDX). Analyses of functional genes within the whole genome of strain TS-2 further revealed the presence of multiple genes predicted to encode laccases/multicopper oxidases that were potentially responsible for Mn(II) oxidation by this strain. The formation of BMO nodules may have functioned to prevent the complete encrustation of fungal hyphae, thereby enabling the control of heavy metal concentrations in their local microenvironments while maintaining hyphal functionality. The present results will expand our knowledge of the physiological and morphological traits of Mn(II)-oxidizing Periconia, which may affect the natural cycle of heavy metals through their immobilization.


Assuntos
Cobre , Hifas , Compostos de Manganês , Óxidos , Hifas/metabolismo , Hifas/crescimento & desenvolvimento , Cobre/metabolismo , Compostos de Manganês/metabolismo , Óxidos/metabolismo , Óxidos/química , Ascomicetos/genética , Ascomicetos/metabolismo , Ascomicetos/química , Oxirredução , Água Subterrânea/microbiologia , Água Subterrânea/química , Filogenia , Sedimentos Geológicos/microbiologia , Microscopia Eletrônica de Varredura , Manganês/metabolismo
8.
PLoS Biol ; 22(6): e3002693, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38905306

RESUMO

Candida albicans is a commensal of the human microbiota that can form biofilms on implanted medical devices. These biofilms are tolerant to antifungals and to the host immune system. To identify novel genes modulating C. albicans biofilm formation, we performed a large-scale screen with 2,454 C. albicans doxycycline-dependent overexpression strains and identified 16 genes whose overexpression significantly hampered biofilm formation. Among those, overexpression of the ZCF15 and ZCF26 paralogs that encode transcription factors and have orthologs only in biofilm-forming species of the Candida clade, caused impaired biofilm formation both in vitro and in vivo. Interestingly, overexpression of ZCF15 impeded biofilm formation without any defect in hyphal growth. Transcript profiling, transcription factor binding, and phenotypic microarray analyses conducted upon overexpression of ZCF15 and ZCF26 demonstrated their role in reprogramming cellular metabolism by regulating central metabolism including glyoxylate and tricarboxylic acid cycle genes. Taken together, this study has identified a new set of biofilm regulators, including ZCF15 and ZCF26, that appear to control biofilm development through their specific role in metabolic remodeling.


Assuntos
Biofilmes , Candida albicans , Proteínas Fúngicas , Regulação Fúngica da Expressão Gênica , Fatores de Transcrição , Biofilmes/crescimento & desenvolvimento , Candida albicans/genética , Candida albicans/metabolismo , Candida albicans/fisiologia , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Animais , Plâncton/metabolismo , Glioxilatos/metabolismo , Perfilação da Expressão Gênica/métodos , Camundongos , Ciclo do Ácido Cítrico , Hifas/metabolismo , Hifas/crescimento & desenvolvimento , Hifas/genética , Candidíase/microbiologia , Reprogramação Metabólica
9.
BMC Microbiol ; 24(1): 227, 2024 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-38937715

RESUMO

This study investigated the influence of bacterial cyclic lipopeptides (LP; surfactins, iturins, fengycins) on microbial interactions. The objective was to investigate whether the presence of bacteria inhibits fungal growth and whether this inhibition is due to the release of bacterial metabolites, particularly LP. Selected endophytic bacterial strains with known plant-growth promoting potential were cultured in the presence of Fusarium oxysporum f.sp. strigae (Fos), which was applied as model fungal organism. The extracellular metabolome of tested bacteria, with a focus on LP, was characterized, and the inhibitory effect of bacterial LP on fungal growth was investigated. The results showed that Bacillus velezensis GB03 and FZB42, as well as B. subtilis BSn5 exhibited the strongest antagonism against Fos. Paraburkholderia phytofirmans PsJN, on the other hand, tended to have a slight, though non-significant growth promotion effect. Crude LP from strains GB03 and FZB42 had the strongest inhibitory effect on Fos, with a significant inhibition of spore germination and damage of the hyphal structure. Liquid chromatography tandem mass spectrometry revealed the production of several variants of iturin, fengycin, and surfactin LP families from strains GB03, FZB42, and BSn5, with varying intensity. Using plate cultures, bacillomycin D fractions were detected in higher abundance in strains GB03, FZB42, and BSn5 in the presence of Fos. Additionally, the presence of Fos in dual plate culture triggered an increase in bacillomycin D production from the Bacillus strains. The study demonstrated the potent antagonistic effect of certain Bacillus strains (i.e., GB03, FZB42, BSn5) on Fos development. Our findings emphasize the crucial role of microbial interactions in shaping the co-existence of microbial assemblages.


Assuntos
Antibiose , Antifúngicos , Bacillus , Fusarium , Lipopeptídeos , Fusarium/efeitos dos fármacos , Fusarium/crescimento & desenvolvimento , Lipopeptídeos/farmacologia , Lipopeptídeos/metabolismo , Bacillus/metabolismo , Antifúngicos/farmacologia , Peptídeos Cíclicos/farmacologia , Interações Microbianas , Burkholderiaceae/crescimento & desenvolvimento , Burkholderiaceae/metabolismo , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento
10.
Commun Biol ; 7(1): 735, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38890525

RESUMO

Utilizing a microfluidic chip with serpentine channels, we inoculated the chip with an agar plug with Neurospora crassa mycelium and successfully captured individual hyphae in channels. For the first time, we report the presence of an autonomous clock in hyphae. Fluorescence of a mCherry reporter gene driven by a clock-controlled gene-2 promoter (ccg-2p) was measured simultaneously along hyphae every half an hour for at least 6 days. We entrained single hyphae to light over a wide range of day lengths, including 6,12, 24, and 36 h days. Hyphae tracked in individual serpentine channels were highly synchronized (K = 0.60-0.78). Furthermore, hyphae also displayed temperature compensation properties, where the oscillation period was stable over a physiological range of temperatures from 24 °C to 30 °C (Q10 = 1.00-1.10). A Clock Tube Model developed could mimic hyphal growth observed in the serpentine chip and provides a mechanism for the stable banding patterns seen in race tubes at the macroscopic scale and synchronization through molecules riding the growth wave in the device.


Assuntos
Hifas , Neurospora crassa , Neurospora crassa/genética , Neurospora crassa/fisiologia , Neurospora crassa/crescimento & desenvolvimento , Hifas/crescimento & desenvolvimento , Hifas/genética , Temperatura , Dispositivos Lab-On-A-Chip , Regulação Fúngica da Expressão Gênica , Relógios Biológicos/genética
11.
Appl Microbiol Biotechnol ; 108(1): 398, 2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-38940906

RESUMO

Grey mould caused by Botrytis cinerea is a devastating disease responsible for large losses to agricultural production, and B. cinerea is a necrotrophic model fungal plant pathogen. Membrane proteins are important targets of fungicides and hotspots in the research and development of fungicide products. Wuyiencin affects the permeability and pathogenicity of B. cinerea, parallel reaction monitoring revealed the association of membrane protein Bcsdr2, and the bacteriostatic mechanism of wuyiencin was elucidated. In the present work, we generated and characterised ΔBcsdr2 deletion and complemented mutant B. cinerea strains. The ΔBcsdr2 deletion mutants exhibited biofilm loss and dissolution, and their functional activity was illustrated by reduced necrotic colonisation on strawberry and grape fruits. Targeted deletion of Bcsdr2 also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted gene complementation. The roles of Bcsdr2 in biofilms and pathogenicity were also supported by quantitative real-time RT-PCR results showing that phosphatidylserine decarboxylase synthesis gene Bcpsd and chitin synthase gene BcCHSV II were downregulated in the early stages of infection for the ΔBcsdr2 strain. The results suggest that Bcsdr2 plays important roles in regulating various cellular processes in B. cinerea. KEY POINTS: • The mechanism of wuyiencin inhibits B. cinerea is closely associated with membrane proteins. • Wuyiencin can downregulate the expression of the membrane protein Bcsdr2 in B. cinerea. • Bcsdr2 is involved in regulating B. cinerea virulence, growth and development.


Assuntos
Biofilmes , Botrytis , Fragaria , Proteínas Fúngicas , Hifas , Proteínas de Membrana , Doenças das Plantas , Botrytis/patogenicidade , Botrytis/genética , Botrytis/crescimento & desenvolvimento , Botrytis/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Virulência , Hifas/crescimento & desenvolvimento , Hifas/efeitos dos fármacos , Doenças das Plantas/microbiologia , Fragaria/microbiologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Vitis/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/genética , Deleção de Genes
12.
Microbiol Spectr ; 12(7): e0341923, 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38842336

RESUMO

Cryptococcus neoformans is a fungal pathogen responsible for >200,000 yearly cases with a mortality as high as 81%. This burden results, in part, from an incomplete understanding of its pathogenesis and ineffective antifungal treatments; hence, there is a pressing need to understand the biology and host interactions of this yeast to develop improved treatments. Protein palmitoylation is important for cryptococcal virulence, and we previously identified the substrates of its main palmitoyl transferase. One of them was encoded by the uncharacterized gene CNAG_02129. In the filamentous fungus Neurospora crassa, a homolog of this gene named hyphal anastomosis protein 13 plays a role in proper cellular communication and filament fusion. In Cryptococcus, cellular communication is essential during mating; therefore, we hypothesized that CNAG_02129, which we named hyphal anastomosis protein 1 (HAM1), may play a role in mating. We found that ham1Δ mutants produce more fusion products during mating, filament more robustly, and exhibit competitive fitness defects under mating and non-mating conditions. Additionally, we found several differences with the major virulence factor, the polysaccharide capsule, that may affect virulence, consistent with prior studies linking virulence to mating. We observed that ham1Δ mutants have decreased capsule attachment and transfer but exhibit higher amounts of exopolysaccharide shedding and biofilm production. Finally, HAM1 expression is significantly lower in mating media relative to non-mating conditions, consistent with it acting as a negative regulator of mating. Understanding the connection between mating and virulence in C. neoformans may open new avenues of investigation into ways to improve the treatment of this disease. IMPORTANCE: Fungal mating is a vital part of the lifecycle of the pathogenic yeast Cryptococcus neoformans. More than just ensuring the propagation of the species, mating allows for sexual reproduction to occur and generates genetic diversity as well as infectious propagules that can invade mammalian hosts. Despite its importance in the biology of this pathogen, we still do not know all of the major players regulating the mating process and if they are involved or impact its pathogenesis. Here, we identified a novel negative regulator of mating that also affects certain cellular characteristics known to be important for virulence. This gene, which we call HAM1, is widely conserved across the cryptococcal family as well as in many pathogenic fungal species. This study will open new avenues of exploration regarding the function of uncharacterized but conserved genes in a variety of pathogenic fungal species and specifically in serotype A of C. neoformans.


Assuntos
Criptococose , Cryptococcus neoformans , Proteínas Fúngicas , Fatores de Virulência , Cryptococcus neoformans/genética , Cryptococcus neoformans/patogenicidade , Cryptococcus neoformans/fisiologia , Cryptococcus neoformans/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Virulência/genética , Criptococose/microbiologia , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Genes Fúngicos Tipo Acasalamento/genética , Fenótipo , Regulação Fúngica da Expressão Gênica , Animais , Hifas/genética , Hifas/crescimento & desenvolvimento , Hifas/metabolismo , Camundongos
14.
Virulence ; 15(1): 2362748, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38860453

RESUMO

Rad6 functions as a ubiquitin-conjugating protein that regulates cellular processes in many fungal species. However, its role in filamentous entomopathogenic fungi remains poorly understood. This study characterizes Rad6 in Beauveria bassiana, a filamentous fungus widely employed as a critical fungicide globally. The results demonstrate a significant association between Rad6 and conidial properties, heat shock response, and UV-B tolerance. Concurrently, the mutant strain exhibited heightened sensitivity to oxidative stress, cell wall interfering agents, DNA damage stress, and prolonged heat shock. Furthermore, the absence of Rad6 significantly extended the median lethal time (LT50) of Galleria mellonella infected by B. bassiana. This delay could be attributed to reduced Pr1 proteases and extracellular cuticle-degrading enzymes, diminished dimorphic transition rates, and dysregulated antioxidant enzymes. Additionally, the absence of Rad6 had a more pronounced effect on genetic information processing, metabolism, and cellular processes under normal conditions. However, its impact was limited to metabolism in oxidative stress. This study offers a comprehensive understanding of the pivotal roles of Rad6 in conidial and hyphal stress tolerance, environmental adaptation, and the pathogenesis of Beauveria bassiana.


Assuntos
Beauveria , Proteínas Fúngicas , Esporos Fúngicos , Animais , Beauveria/patogenicidade , Beauveria/genética , Beauveria/fisiologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Resposta ao Choque Térmico , Hifas/crescimento & desenvolvimento , Mariposas/microbiologia , Estresse Oxidativo , Esporos Fúngicos/genética , Estresse Fisiológico , Enzimas de Conjugação de Ubiquitina/metabolismo , Enzimas de Conjugação de Ubiquitina/genética , Virulência
15.
Microbiol Res ; 286: 127792, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38852300

RESUMO

Botrytis cinerea is the phytopathogenic fungus responsible for the gray mold disease that affects crops worldwide. Essential oils (EOs) have emerged as a sustainable tool to reduce the adverse impact of synthetic fungicides. Nevertheless, the scarce information about the physiological mechanism action and the limitations to applying EOs has restricted its use. This study focused on elucidating the physiological action mechanisms and prospection of lipid nanoparticles to apply EO of Mentha piperita. The results showed that the EO of M. piperita at 500, 700, and 900 µL L-1 inhibited the mycelial growth at 100 %. The inhibition of spore germination of B. cinerea reached 31.43 % at 900 µL L-1. The EO of M. piperita decreased the dry weight and increased pH, electrical conductivity, and cellular material absorbing OD260 nm of cultures of B. cinerea. The fluorescence technique revealed that EO reduced hyphae width, mitochondrial activity, and viability, and increased ROS production. The formulation of EO of M. piperita loaded- solid lipid nanoparticles (SLN) at 500, 700, and 900 µL L-1 had particle size ∼ 200 nm, polydispersity index < 0.2, and stability. Also, the thermogravimetric analysis indicated that the EO of M. piperita-loaded SLN has great thermal stability at 50 °C. EO of M. piperita-loaded SLN reduced the mycelial growth of B. cinerea by 70 %, while SLN formulation (without EO) reached 42 % inhibition. These results supported that EO of M. piperita-loaded SLN is a sustainable tool for reducing the disease produced by B. cinerea.


Assuntos
Botrytis , Mentha piperita , Nanopartículas , Óleos Voláteis , Esporos Fúngicos , Botrytis/efeitos dos fármacos , Botrytis/crescimento & desenvolvimento , Óleos Voláteis/farmacologia , Óleos Voláteis/química , Nanopartículas/química , Mentha piperita/química , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento , Fungicidas Industriais/farmacologia , Fungicidas Industriais/química , Micélio/efeitos dos fármacos , Micélio/crescimento & desenvolvimento , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia , Lipídeos/química , Lipídeos/farmacologia , Tamanho da Partícula , Espécies Reativas de Oxigênio/metabolismo , Óleos de Plantas/farmacologia , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Antifúngicos/farmacologia , Lipossomos
16.
Microbiol Res ; 286: 127789, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38870619

RESUMO

Plants have developed intricate immune mechanisms to impede Phytophthora colonization. In response, Phytophthora secretes RxLR effector proteins that disrupt plant defense and promote infection. The specific molecular interactions through which Phytophthora RxLR effectors undermine plant immunity, however, remain inadequately defined. In this study, we delineate the role of the nuclear-localized RxLR effector PcAvh87, which is pivotal for the full virulence of Phytophthora cinnamomi. Gene expression analysis indicates that PcAvh87 expression is significantly upregulated during the initial infection stages, interacting with the immune responses triggered by the elicitin protein INF1 and pro-apoptotic protein BAX. Utilizing PEG/CaCl2-mediated protoplast transformation and CRISPR/Cas9-mediated gene editing, we generated PcAvh87 knockout mutants, which demonstrated compromised hyphal growth, sporangium development, and zoospore release, along with a marked reduction in pathogenicity. This underscores PcAvh87's crucial role as a virulence determinant. Notably, PcAvh87, conserved across the Phytophthora genus, was found to modulate the activity of plant immune protein 113, thereby attenuating plant immune responses. This implies that the PcAvh87-mediated regulatory mechanism could be a common strategy in Phytophthora species to manipulate plant immunity. Our findings highlight the multifaceted roles of PcAvh87 in promoting P. cinnamomi infection, including its involvement in sporangia production, mycelial growth, and the targeting of plant immune proteins to enhance pathogen virulence.


Assuntos
Morte Celular , Phytophthora , Doenças das Plantas , Imunidade Vegetal , Phytophthora/patogenicidade , Phytophthora/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Virulência , Fatores de Virulência/genética , Núcleo Celular/metabolismo , Interações Hospedeiro-Patógeno , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/imunologia , Nicotiana/microbiologia , Nicotiana/imunologia , Hifas/genética , Hifas/crescimento & desenvolvimento , Hifas/imunologia
17.
Microb Pathog ; 193: 106750, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38906491

RESUMO

The antifungal activity of Serratia plymuthica CCGG2742, a bacterial strain isolated from grapes berries skin, against a phytopathogenic fungus isolated from blueberries was evaluated in vitro and in vivo. In order to characterize the wild fungal isolate, phylogenetic analysis using concatenated DNA sequences from the RPB2 and TEF1 genes and of the ITS region was performed, allowing the identification of the fungal isolate that was called Alternaria tenuissima CC17. Hyphae morphology, mycelium ultrastructure, conidia and reproductive structures were in agreement with the phylogenetic analysis. The antifungal activity of the S. plymuthica strain was dependent on the composition of the culture medium. The greatest inhibition of mycelial growth of A. tenuissima CC17 by S. plymuthica CCGG2742 was observed on YTS medium, which lacks of an easily assimilable carbon source. Fungal growth medium supplemented with 50 % of bacterial supernatant decreased the conidia germination of A. tenuissima CC17 up to 32 %. Preventive applications of S. plymuthica CCGG2742 to blueberries and tomato leaves at conidia:bacteria ratio of 1:100, protected in 77.8 ± 4.6 % and 98.2 ± 0.6 % to blueberries and tomato leaves from infection caused by A. tenuissima CC17, respectively. To the best of our knowledge, this is the first report on the antifungal activity of S. plymuthica against A. tenuissima, which could be used as a biological control agent of plant diseases caused by this fungal species. In addition, the results of this work could be a starting point to attribute the real importance of A. tenuissima as a pathogen of blueberries in Chile, which until now had been considered almost exclusively to A. alternata. Likewise, this research could be relevant to start developing highly effective strategies based on S. plymuthica CCGG2742 for the control of this important phytopathogenic fungus.


Assuntos
Alternaria , Antibiose , Filogenia , Doenças das Plantas , Serratia , Esporos Fúngicos , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Alternaria/crescimento & desenvolvimento , Alternaria/genética , Serratia/genética , Serratia/fisiologia , Esporos Fúngicos/crescimento & desenvolvimento , Micélio/crescimento & desenvolvimento , Antifúngicos/farmacologia , Solanum lycopersicum/microbiologia , Hifas/crescimento & desenvolvimento , Meios de Cultura/química , Folhas de Planta/microbiologia , Vitis/microbiologia
18.
Front Cell Infect Microbiol ; 14: 1414618, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38903941

RESUMO

Candida species comprise a ubiquitous pathogenic fungal genus responsible for causing candidiasis. They are one of the primary causatives of several mucosal and systemic infections in humans and can survive in various environments. In this study, we investigated the antifungal, anti-biofilm, and anti-hyphal effects of six N-substituted phthalimides against three Candida species. Of the derivatives, N-butylphthalimide (NBP) was the most potent, with a minimum inhibitory concentration (MIC) of 100 µg/ml and which dose-dependently inhibited biofilm at sub-inhibitory concentrations (10-50 µg/ml) in both the fluconazole-resistant and fluconazole-sensitive Candida albicans and Candida parapsilosis. NBP also effectively inhibited biofilm formation in other pathogens including uropathogenic Escherichia coli, Staphylococcus epidermidis, Staphylococcus aureus, and Vibrio parahaemolyticus, along with the polymicrobial biofilms of S. epidermidis and C. albicans. NBP markedly inhibited the hyphal formation and cell aggregation of C. albicans and altered its colony morphology in a dose-dependent manner. Gene expression analysis showed that NBP significantly downregulated the expression of important hyphal- and biofilm-associated genes, i.e., ECE1, HWP1, and UME6, upon treatment. NBP also exhibited mild toxicity at concentrations ranging from 2 to 20 µg/ml in a nematode model. Therefore, this study suggests that NBP has anti-biofilm and antifungal potential against various Candida strains.


Assuntos
Antifúngicos , Biofilmes , Candida albicans , Hifas , Testes de Sensibilidade Microbiana , Ftalimidas , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Antifúngicos/farmacologia , Ftalimidas/farmacologia , Candida albicans/efeitos dos fármacos , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Candida/efeitos dos fármacos , Candidíase/microbiologia , Candidíase/tratamento farmacológico , Animais , Humanos , Candida parapsilosis/efeitos dos fármacos , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fluconazol/farmacologia
19.
Sheng Wu Gong Cheng Xue Bao ; 40(6): 1776-1791, 2024 Jun 25.
Artigo em Chinês | MEDLINE | ID: mdl-38914491

RESUMO

Filamentous fungi are a group of eukaryotic microorganisms widely found in nature. Some filamentous fungi have been developed as "cell factories" and extensively used for the production of recombinant proteins, organic acids, and secondary metabolites due to their strong protein secretion capabilities or effective synthesis of many natural products. The growth morphology of filamentous fungi significantly influences the quality and quantity of fermented products. Previous research conducted by the authors' group revealed that an increase in hyphal branches leads to enhanced protein secretion during liquid fermentation. With the development of morphological engineering of filamentous fungi, an increasing number of studies have focused on modifying fungal mycelium morphology to improve the yield of target metabolites during fermentation. While there have been a few reviews on the relationship between fungal fermentation morphology and productivity, research in this area is rapidly developing and requires updates. The paper presents a comprehensive review of domestic and international research reports, along with the authors' own research findings, to systematically review the morphological patterns of filamentous fungi, the impact of fungal morphology on industrial fermentation, as well as methods and strategies for regulating mycelial morphology. The aim of this review is to enhance the understanding of relevant domestic scholars regarding the morphological development of filamentous fungi and provide ideas for the rational engineering of fungal strains suitable for industrial fermentation.


Assuntos
Fermentação , Fungos , Micélio , Fungos/genética , Fungos/metabolismo , Micélio/genética , Micélio/metabolismo , Micélio/crescimento & desenvolvimento , Microbiologia Industrial , Engenharia Genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Hifas/genética , Hifas/crescimento & desenvolvimento
20.
Microbiome ; 12(1): 83, 2024 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-38725008

RESUMO

BACKGROUND: Fungi and bacteria coexist in a wide variety of environments, and their interactions are now recognized as the norm in most agroecosystems. These microbial communities harbor keystone taxa, which facilitate connectivity between fungal and bacterial communities, influencing their composition and functions. The roots of most plants are associated with arbuscular mycorrhizal (AM) fungi, which develop dense networks of hyphae in the soil. The surface of these hyphae (called the hyphosphere) is the region where multiple interactions with microbial communities can occur, e.g., exchanging or responding to each other's metabolites. However, the presence and importance of keystone taxa in the AM fungal hyphosphere remain largely unknown. RESULTS: Here, we used in vitro and pot cultivation systems of AM fungi to investigate whether certain keystone bacteria were able to shape the microbial communities growing in the hyphosphere and potentially improved the fitness of the AM fungal host. Based on various AM fungi, soil leachates, and synthetic microbial communities, we found that under organic phosphorus (P) conditions, AM fungi could selectively recruit bacteria that enhanced their P nutrition and competed with less P-mobilizing bacteria. Specifically, we observed a privileged interaction between the isolate Streptomyces sp. D1 and AM fungi of the genus Rhizophagus, where (1) the carbon compounds exuded by the fungus were acquired by the bacterium which could mineralize organic P and (2) the in vitro culturable bacterial community residing on the surface of hyphae was in part regulated by Streptomyces sp. D1, primarily by inhibiting the bacteria with weak P-mineralizing ability, thereby enhancing AM fungi to acquire P. CONCLUSIONS: This work highlights the multi-functionality of the keystone bacteria Streptomyces sp. D1 in fungal-bacteria and bacterial-bacterial interactions at the hyphal surface of AM fungi. Video Abstract.


Assuntos
Hifas , Microbiota , Micorrizas , Raízes de Plantas , Microbiologia do Solo , Streptomyces , Micorrizas/fisiologia , Micorrizas/classificação , Streptomyces/classificação , Streptomyces/isolamento & purificação , Streptomyces/genética , Streptomyces/fisiologia , Hifas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , Fósforo/metabolismo , Interações Microbianas/fisiologia , Solo/química , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo
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