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1.
PLoS One ; 15(3): e0229738, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32160222

RESUMO

Ultrasound has many uses, such as in medical imaging, monitoring of crystallization, characterization of emulsions and suspensions, and disruption of cell membranes in the food industry. It can also affect microbial cells by promoting or slowing their growth and increasing the production of some metabolites. However, the exact mechanism explaining the effect of ultrasound has not been identified yet. Most equipment employed to study the effect of ultrasound on microorganisms has been designed for other applications and then only slightly modified. This results in limited control over ultrasound frequency and input power, or pressure distribution in the reactor. The present study aimed to obtain a well-defined reactor by simulating the pressure distribution of a sonobioreactor. Specifically, we optimized a sonotrode to match the bottle frequency and compared it to measured results to verify the accuracy of the simulation. The measured pressure distribution spectrum presented the same overall trend as the simulated spectrum. However, the peaks were much less intense, likely due to non-linear events such as the collapse of cavitation bubbles. To test the application of the sonobioreactor in biological systems, two biotechnologically interesting microorganisms were assessed: an electroactive bacterium, Geobacter sulfurreducens, and a lignocellulose-degrading fungus, Fusarium oxysporum. Sonication resulted in increased malate production by G. sulfurreducens, but no major effect on growth. In comparison, morphology and growth of F. oxysporum were more sensitive to ultrasound intensity. Despite considerable morphological changes at 4 W input power, the growth rate was not adversely affected; however, at 12 W, growth was nearly halted. The above findings indicate that the novel sonobioreactor provides an effective tool for studying the impact of ultrasound on microorganisms.


Assuntos
Reatores Biológicos/microbiologia , Fusarium/crescimento & desenvolvimento , Geobacter/crescimento & desenvolvimento , Análise Numérica Assistida por Computador , Sonicação , Calorimetria , Simulação por Computador , Fusarium/ultraestrutura , Geobacter/metabolismo , Malatos/metabolismo , Metaboloma , Pressão , Vibração
2.
J Hazard Mater ; 388: 122063, 2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-31972432

RESUMO

Silver nanoparticles (AgNPs) have been widely used in various fields due to their antimicrobial activities. However, the antimicrobial mechanisms of AgNPs against fungi, especially on transcriptional level, are still unclear. In this study, the inhibitory property of AgNPs against Fusarium solani species complex was investigated. Transmission electron microscopes were used to observe the alterations in morphology and cellular structure of fungal hyphae treated with AgNPs. Disturbances in the cell walls and membranes, as well as empty space in the cytoplasm were observed. The transcriptome sequencing of F. solani species complex mycelia was performed using the Illumina NextSeq 500 ribonucleic acid sequencing (RNA-Seq) platform. In the RNA-Seq study, AgNPs treatment resulted in 2503 differentially expressed genes (DEGs). Gene Ontology (GO) analysis revealed that the DEGs were mainly involved in 6 different terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis also revealed that energy and substance metabolism, signal transduction and genetic information processing were the most highly enriched pathways for these DEGs. In addition, RNA-seq results were validated by quantitative polymerase chain reactions (qPCRs). Our findings enhanced the understanding of the antifungal activities of AgNPs and the underlying molecular mechanisms, and provided a new perspective for investigating this novel antifungal agent.


Assuntos
Antifúngicos/administração & dosagem , Fusarium/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Prata/administração & dosagem , Fusarium/genética , Fusarium/crescimento & desenvolvimento , Fusarium/ultraestrutura , Perfilação da Expressão Gênica , Microscopia Eletrônica de Transmissão , Transcriptoma
3.
Transbound Emerg Dis ; 67(2): 555-563, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31539213

RESUMO

Zoonotic Fusarium oxysporum infection was identified in captive-reared fingerlings of golden mahseer, Tor putitora (Hamilton, 1822) from the central Himalayan regions, India. Initially, fingerlings of T. putitora (mean length 10.8 ± 0.002 and weight 18.58 ± 0.054 g) were observed with cottony mass like growth completely covering the dorsal and caudal fins. The infected fingerlings were showing clinical signs such as sluggish, erratic movement, gasping, flared operculum and settling at one corner of the rearing tanks. The microscopic observation of 8-day old culture of cottony mass like growth showed the presence of septate macroconidia, randomly spread microconidia and chlamydospores in short-chain. From sequence analysis of ITS amplified fragment, the isolate was identified as Fusarium oxysporum, TPFCF 214 (MH464266.1) and clustered with F. oxysporum, strain NRRL 43504 (EF453107.1) and F. oxysporum, strain 20736 (JX 270150.1) isolated from the human in phylogenetic tree. An experimental infection of healthy golden mahseer fingerlings with 20 µl of F. oxysporum spore suspension (2.5 × 109 spore ml-1 ) showed the development of lesion 6-dpi at the site of injection. Experimental trial on EPC-2 cell culture recorded detachment in the monolayer, clumping and shrinking of the cell line 6-8 dpi with a spore suspension of F. oxysporum, TPFCF 214 (5.68 × 102 cell/ml). From the severity of its infection, there is a chance that F. oxysporum may emerge as pathogenically and pose a significant health risk on captive-reared golden mahseer in other Asian countries and world. As Fusarium solani and F. oxysporum are known to cause invasive fusariosis in human especially in immunocompromised patients, localized infection in immunocompetent individuals as well as osteomyelitis, arthritis, otitis, sinusitis and brain abscess, the global fish farmers, handlers and aquaculturist need to be aware of possible health hazards caused by Fusarium spp. and should adopt proper fish health management and animal husbandry practice to control the infection of Fusarium in culture environment.


Assuntos
Cyprinidae/microbiologia , Doenças dos Peixes/epidemiologia , Fusariose/epidemiologia , Fusarium/isolamento & purificação , Animais , Linhagem Celular , Doenças dos Peixes/microbiologia , Doenças dos Peixes/mortalidade , Fusariose/microbiologia , Fusariose/mortalidade , Fusarium/genética , Fusarium/fisiologia , Fusarium/ultraestrutura , Humanos , Índia/epidemiologia , Filogenia , Prevalência , Esporos Fúngicos , Zoonoses
4.
Int J Nanomedicine ; 14: 7975-7985, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31632012

RESUMO

BACKGROUND: Multidrug-resistant bacteria such as extended-spectrum beta-lactamase (ESBL), Enterobacteriaceae, and methicillin-resistant Staphylococcus aureus (MRSA) pose a challenge to the human health care system. MRSA is among the major causes of hospital-acquired and community infections. METHODS: Therefore, in the present study, we evaluated the antibacterial activity of silver nanoparticles synthesized by Fusarium oxysporum (AgNPbio) in combination with simvastatin against reference and multidrug-resistant bacterial strains. RESULTS: Simvastatin showed a minimal inhibitory concentration (MIC) ranging from 0.062 to 0.25 mg mL-1 against MRSA. AgNPbio with a size of 77.68± 33.95 nm and zeta potential -34.6 ± 12.7 mV showed an MIC of 0.212 mg mL-1 against S. aureus including MRSA strains. The checkerboard assay and time-kill curves exhibited a synergistic effect of the simvastatin-AgNPbio combination on antibacterial activity against MRSA strains. The combination of simvastatin and AgNPbio demonstrated antibacterial activity against Escherichia coli producing ESBL. Scanning electron microscopy showed the formation of cell surface protrusions after treatment with AgNPbio and the formation of a large amorphous mass after treatment with simvastatin, both in MRSA. CONCLUSION: Our results indicate that the combination of AgNPbio and simvastatin could be a great future alternative in the control of bacterial infections, where, when combined with simvastatin, smaller doses of AgNPbio are required, with the same antibacterial activity.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Fusarium/metabolismo , Nanopartículas Metálicas/química , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Prata/farmacologia , Sinvastatina/farmacologia , Morte Celular/efeitos dos fármacos , Sinergismo Farmacológico , Eritrócitos/efeitos dos fármacos , Fusarium/efeitos dos fármacos , Fusarium/ultraestrutura , Humanos , Nanopartículas Metálicas/ultraestrutura , Staphylococcus aureus Resistente à Meticilina/ultraestrutura , Testes de Sensibilidade Microbiana
5.
Toxins (Basel) ; 11(5)2019 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-31137632

RESUMO

Fusarium graminearum is a notorious pathogen that causes Fusarium head blight (FHB) in cereal crops. It produces secondary metabolites, such as deoxynivalenol, diminishing grain quality and leading to lesser crop yield. Many strategies have been developed to combat this pathogenic fungus; however, considering the lack of resistant cultivars and likelihood of environmental hazards upon using chemical pesticides, efforts have shifted toward the biocontrol of plant diseases, which is a sustainable and eco-friendly approach. Fengycin, derived from Bacillus amyloliquefaciens FZB42, was purified from the crude extract by HPLC and further analyzed by MALDI-TOF-MS. Its application resulted in structural deformations in fungal hyphae, as observed via scanning electron microscopy. In planta experiment revealed the ability of fengycin to suppress F. graminearum growth and highlighted its capacity to combat disease incidence. Fengycin significantly suppressed F. graminearum, and also reduced the deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), and zearalenone (ZEN) production in infected grains. To conclude, we report that fengycin produced by B. amyloliquefaciens FZB42 has potential as a biocontrol agent against F. graminearum and can also inhibit the mycotoxins produced by this fungus.


Assuntos
Bacillus amyloliquefaciens/metabolismo , Agentes de Controle Biológico/farmacologia , Fusarium/efeitos dos fármacos , Lipopeptídeos/farmacologia , Micotoxinas/biossíntese , Bacillus amyloliquefaciens/genética , Agentes de Controle Biológico/metabolismo , Fusarium/crescimento & desenvolvimento , Fusarium/metabolismo , Fusarium/ultraestrutura , Lipopeptídeos/metabolismo , Microscopia Eletrônica , Triticum/microbiologia
6.
Food Chem ; 289: 278-284, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-30955613

RESUMO

Fusarium rot of muskmelon is a common and frequently-occurring postharvest disease, which leads to quality deterioration and neosolaniol (NEO) contamination. New strategies to control postharvest decay and reduce NEO contamination are of paramount importance. The effects of acetylsalicylic acid (ASA) treatment on the growth of Fusarium sulphureum in vitro, and Fusarium rot development and NEO accumulation in fruits inoculated with F. sulphureum in vivo were investigated. The results showed that ASA inhibited the growth of F. sulphureum, evident morphological and major cellular changes were observed under the microscope. In vivo testing showed that 3.2 mg/mL ASA significantly suppressed Fusarium rot development and NEO accumulation after 6 and 8 d of pathogen inoculation. Meanwhile, Tri gene expressions involved in NEO biosynthesis were down-regulated after treatment. Taken together, ASA treatment not only reduced Fusarium rot development by inhibiting the growth of F. sulphureum, but decreased NEO accumulation by suppressing NEO biosynthesis pathway.


Assuntos
Aspirina/farmacologia , Cucurbitaceae/química , Tricotecenos/metabolismo , Cucurbitaceae/metabolismo , Cucurbitaceae/microbiologia , Frutas/química , Frutas/metabolismo , Frutas/microbiologia , Fusarium/efeitos dos fármacos , Fusarium/crescimento & desenvolvimento , Fusarium/metabolismo , Fusarium/ultraestrutura , Doenças das Plantas/prevenção & controle , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA de Plantas/genética , RNA de Plantas/metabolismo , Tricotecenos/química
7.
Int J Mol Sci ; 20(5)2019 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-30813526

RESUMO

In this study, two Burkholderia strains, strain KNU17BI2 and strain KNU17BI3, were isolated from maize rhizospheric soil, South Korea. The 16S rRNA gene and multilocus sequence analysis and typing (MLSA-MLST) were used for the identification of the studied strains. Strain KNU17BI2, which belonged to Burkholderia cenocepacia, was of a novel sequence type (ST) designated ST-1538, while strain KNU17BI3 had a similar allelic profile with the seven loci of Burkholderia contaminans strain LMG 23361. The strains were evaluated in vitro for their specific plant growth promoting (PGP) traits, such as zinc solubilization, phosphate solubilization, ammonia production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, indole acetic acid (IAA) production, siderophore, and hydrolytic enzyme activity. Interestingly, the strains exhibited a positive effect on all of the tested parameters. The strains also showed broad-spectrum antifungal activity against economically important phytopathogens in the dual culture assay. Furthermore, the strains were evaluated under greenhouse conditions for their in vivo effect to promote plant growth and to suppress the root rot of maize that is caused by Fusarium temperatum on four Korean maize cultivars. The results of the greenhouse study revealed that both of the strains were promising to significantly suppress fusarium root rot and enhance plant growth promotion on the four maize cultivars. This study, for the first time, reported in vitro antifungal potential of B. cenocepacia of novel ST against economically important plant pathogens viz., F. temperatum, Fusarium graminearum, Fusarium moniliforme, Fusarium oxysporum f.sp. melonis, Fusarium subglutinans, Phytophthora drechsleri, and Stemphylium lycopersici. This is also the first report of zinc solubilization by B. cenocepacia. Moreover, the present research work reports, for the first time, about the potential of B. cenocepacia and B. contaminans to control the root rot of maize that is caused by F. temperatum. Therefore, we recommend further studies to precisely identify the bioactive chemical compounds behind such activities that would be novel sources of natural products for biological control and plant growth promotion of different crops.


Assuntos
Burkholderia/classificação , Fusarium/fisiologia , Tipagem de Sequências Multilocus , Controle Biológico de Vetores , Doenças das Plantas/microbiologia , Zea mays/microbiologia , Alelos , Antifúngicos/farmacologia , Sequência de Bases , Biomassa , Burkholderia/isolamento & purificação , Fusarium/efeitos dos fármacos , Fusarium/patogenicidade , Fusarium/ultraestrutura , Ácidos Indolacéticos/metabolismo , Testes de Sensibilidade Microbiana , Fosfatos/metabolismo , Filogenia , Desenvolvimento Vegetal/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/microbiologia , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/microbiologia , RNA Ribossômico 16S/genética , Solubilidade , Triptofano/metabolismo , Virulência/efeitos dos fármacos , Zinco/metabolismo
8.
New Phytol ; 223(1): 412-429, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30767239

RESUMO

Lipid droplets (LDs) control lipid metabolism in eukaryotic cells in general. However, the biogenesis regulation and biological functions of LDs are largely unknown in pathogenic fungi. Rapamycin treatment results in a significant increase of LD biogenesis in Fusarium graminearum. Molecular mechanisms of the target of rapamycin (TOR) pathway in regulating LD biogenesis and the functions of LD in virulence of F. graminearum were investigated in depth by combining genetic, cytological and phenotypic strategies. TOR in Fusarium graminearum (FgTOR) inhibition by rapamycin induces LD biogenesis through the FgPpg1/Sit4 signaling branch. FgPpg1 promotes phosphorylation of protein phosphatase FgNem1 by the protein kinase FgCak1. The phosphorylated FgNem1 dephosphorylates the phosphatidate phosphatase FgPah1. Dephosphorylated FgPah1 is active and stimulates LD biogenesis. Moreover, deletion of FgNem1/Spo7 or FgPah1 leads to serious defects in vegetative growth, sexual development and virulence. The results of this study provide novel insights into the regulatory mechanism and biological functions of the LDs in the devastating pathogenic fungus F. graminearum.


Assuntos
Proteínas Fúngicas/metabolismo , Fusarium/crescimento & desenvolvimento , Fusarium/patogenicidade , Gotículas Lipídicas/metabolismo , Transdução de Sinais , Fusarium/enzimologia , Fusarium/ultraestrutura , Gotículas Lipídicas/efeitos dos fármacos , Gotículas Lipídicas/ultraestrutura , Fosforilação/efeitos dos fármacos , Ligação Proteica/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia , Tricotecenos/metabolismo , Virulência/efeitos dos fármacos
9.
Fungal Genet Biol ; 124: 73-77, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30579887

RESUMO

Trichothecene mycotoxin synthesis in the phytopathogen Fusarium graminearum involves primarily endoplasmic reticulum (ER)-localized enzymes of the mevalonate- and trichothecene biosynthetic pathways. Two exceptions are 3-hydroxy-3-methylglutaryl CoA synthase (Hms1) and trichodiene synthase (Tri5), which are known cytosolic enzymes. Using 3D structured illumination microscopy (3D SIM), GFP-tagged Tri5 and Hms1 were tested for preferential localization in the cytosol proximal to the ER. Tri5 protein was significantly enriched in cytosolic regions within 500 nm of the ER, but Hms1 was not. Spatial organization of enzymes in the cytosol has potential relevance for pathway efficiency and metabolic engineering in fungi and other organisms.


Assuntos
Carbono-Carbono Liases/metabolismo , Fusarium/enzimologia , Citosol/metabolismo , Retículo Endoplasmático Liso/metabolismo , Retículo Endoplasmático Liso/ultraestrutura , Fusarium/ultraestrutura , Redes e Vias Metabólicas , Microscopia/métodos , Micotoxinas/metabolismo , Nanopartículas
10.
Biochim Biophys Acta Biomembr ; 1861(2): 532-539, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30391317

RESUMO

P852, a novel cyclic peptide isolated from Bacillus amyloliquefaciens L-H15, showed potent antifungal activity against several major plant fungal pathogens including Fusarium oxysporum. To elucidate the antifungal mechanism, the impact of P852 on the cell morphology and membrane permeabilization of F. oxysporum was studied. By applying electron microscopy and fluorescent techniques, we showed that P852 treatment caused the morphological change of F. oxysporum cells and disrupted its cell structure, including formation of blebs, broken hyphae, deformation of membrane, intracellular organization disruption, pore formation, and cell lysis. Our findings provide insights into the mode of action of P852, which laying a foundation to develop P852 as a novel antifungal agent to control plant fungal pathogens.


Assuntos
Antifúngicos/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Fusarium/citologia , Peptídeos/farmacologia , Anfotericina B/farmacologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , DNA Fúngico/metabolismo , Polarização de Fluorescência , Fusarium/efeitos dos fármacos , Fusarium/ultraestrutura , Hifas/citologia , Hifas/efeitos dos fármacos , Hifas/ultraestrutura , Concentração Inibidora 50 , Cinética , Fluidez de Membrana/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Compostos Orgânicos/metabolismo
11.
Pestic Biochem Physiol ; 152: 98-105, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30497718

RESUMO

Fusarium fujikuroi is the primary causal agent of rice bakanae disease. Fluazinam is a protective dinitroaniline fungicide which could interrupt the fungal cell's energy production. Little is known about the effects of fluazinam on F. fujikuroi. In this study, baseline sensitivity of F. fujikuroi to fluazinam was determined using 103 isolates collected from diseased young rice of different fields in Shaoxing of Zhejiang Province and Huaian of Jiangsu Province of China in 2016. The EC50 values of fluazinam on inhibiting mycelial growth against 103 isolates of F. fujikuroi ranged from 0.0621 to 0.5446 µg/mL with the average value of 0.2038 ±â€¯0.0099 µg/mL (mean ±â€¯standard error). The EC50 values of fluazinam on suppressing conidium germination against 103 isolates of F. fujikuroi ranged from 0.1006 to 0.9763 µg/mL with the mean value of 0.3552 ±â€¯0.0181 µg/mL. Treated with fluazinam, hyphae of F. fujikuroi were contorted, offshoot of top mycelia increased, conidial production descreased significantly and exopolysaccharide (EPS) content did not change significantly while peroxidase (POD) activity significantly decreased. Meanwhile, cell membrane permeability increased after treated with fluazinam. The analysis of cell ultrastructure indicated that fluazinam could damage the membrane structure of F. fujikuroi and cause a large number of vacuoles formed. In addition, fluazinam did not affect germination rate, plant height and fresh weight of rice, which indicated that fluazinam was safe to rice. All the results indicated that fluazinam had strong antifungal activity against F. fujikuroi and a potential application in controlling rice bakanae disease. These results will provide useful information for management of rice bakanae disease caused by F. fujikuroi and further increase our understanding about the mode of action of fluazinam against F. fujikuroi and other phytopathogens.


Assuntos
Aminopiridinas/farmacologia , Fungicidas Industriais/farmacologia , Fusarium/efeitos dos fármacos , Oryza/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Polissacarídeos Fúngicos/metabolismo , Fusarium/fisiologia , Fusarium/ultraestrutura , Micélio/efeitos dos fármacos , Micélio/fisiologia , Micélio/ultraestrutura , Oryza/crescimento & desenvolvimento , Peroxidase/metabolismo
12.
New Phytol ; 219(4): 1447-1466, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29932228

RESUMO

ATP-binding cassette (ABC) transporters act mainly to transport compounds across cellular membranes and are important for diverse biological processes. However, their roles in pathogenesis have not been well-characterized in Fusarium graminearum. Sixty F. graminearum ABC protein genes were functionally characterized. Among them, FgArb1 regulates normal growth and importantly is essential for pathogenicity. Thus, the regulatory mechanisms of FgArb1 in pathogenicity were analyzed in this study. FgArb1 interacts with the mitogen-activated protein kinase (MAPK) FgSte7, and partially modulates plant penetration by regulating the phosphorylation of FgGpmk1 (the downstream kinase of FgSte7). The FgArb1 mutant exhibited dramatically reduced infective growth within wounded host tissues, likely resulting from its increased sensitivity to oxidative stresses, defective cell wall integrity and reduced deoxynivalenol (DON) production. FgArb1 also is important for the production of sexual and asexual spores that are important propagules for plant infection. In addition, FgArb1 is involved in the regulation of protein biosynthesis through impeding nuclear export of small ribosomal subunit. Finally, acetylation modification at sites K28, K65, K341 and K525 in FgArb1 is required for its biological functions. Taken together, results of this study provide a novel insight into functions of the ABC transporter in fungal pathogenesis.


Assuntos
Trifosfato de Adenosina/metabolismo , Proteínas Fúngicas/metabolismo , Fusarium/crescimento & desenvolvimento , Fusarium/patogenicidade , Acetilação , Transporte Ativo do Núcleo Celular , Núcleo Celular/metabolismo , Parede Celular/metabolismo , Parede Celular/ultraestrutura , Fusarium/ultraestrutura , Lisina/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Modelos Biológicos , Estresse Oxidativo , Subunidades Ribossômicas Menores de Eucariotos/metabolismo , Tricotecenos/metabolismo , Triticum/microbiologia , Triticum/ultraestrutura
13.
Antonie Van Leeuwenhoek ; 111(10): 1785-1805, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29569107

RESUMO

Two new species in the Fusarium solani species complex (FSSC) are described and introduced. The new taxa are represented by German isolates CBS 142481 and CBS 142480 collected from commercial yard waste compost and vascular tissue of a wilting branch of hibiscus, respectively. The phylogenetic relationships of the collected strains to one another and within the FSSC were evaluated based on DNA sequences of 6 gene loci. Due to the limited sequence data available for reference strains in GenBank, however, a multi-gene phylogenetic analysis included partial sequences for the internal transcribed spacer region and intervening 5.8S nrRNA gene (ITS), translation elongation factor 1-alpha (tef1) and the RNA polymerase II second largest subunit (rpb2). Morphological and molecular phylogenetic data independently showed that these strains are distinct populations of the FSSC, nested within Clade 3. Thus, we introduce Fusarium stercicola and Fusarium witzenhausenense as novel species in the complex. In addition, 19 plant species of 7 legume genera were evaluated for their potential to host the newly described taxa. Eighteen plant species were successfully colonized, with 6 and 9 of these being symptomatic hosts for F. stercicola and F. witzenhausenense, respectively. As plants of the family Fabaceae are very distant to the originally sourced material from which the new taxa were recovered, our results suggest that F. stercicola and F. witzenhausenense are not host-specific and are ecologically fit to sustain stable populations in variety of habitats.


Assuntos
Microbiologia Ambiental , Fusarium/classificação , Hibiscus/microbiologia , DNA Fúngico , Fusarium/genética , Fusarium/isolamento & purificação , Fusarium/ultraestrutura , Especificidade de Hospedeiro , Tipagem de Sequências Multilocus , Fenótipo , Filogenia , Esporos Fúngicos
14.
Curr Microbiol ; 74(11): 1301-1305, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28779356

RESUMO

Polyunsaturated fatty acid helps to prevent diseases like cardiovascular, inflammation, and cognitive abilities for developmental disorders. The main objective of this research is the screening of polyunsaturated fatty acid producing fungi from soil samples of mangrove from the seashore coastal areas in India. Fusarium verticillioides species showed the presence of saturated and unsaturated fatty acid in the starch yeast-extract medium. Among the representative isolate, F. verticillioides NKF1 was found to grow in a YEP broth medium and produce the maximum lipid. The gas chromatography was used to identify the fatty acids present in fungal strain. Saturated fatty acid such as palmitic acid (C16:0) 0.14/100 g, stearic acid (C18:0) 0.09/100 g, and monounsaturated fatty acid such as oleic acid (C18:1) 0.08/100 g and polyunsaturated fatty acid such as linolenic acid (C18:3ω3) 0.08/100 g were present in significant amount in the fungal strain. Fungal strain F. verticillioides NKF1 was characterized by SEM and molecular characterization by 18S rRNA. The internal transcribed spacer (ITS) sequences were analyzed by 18S rRNA and ITS4 sequences of related fungi were sequenced, and then the data were compared with NCBI database. This newly isolated F. verticillioides NKF1 was found to be a promising culture for the development of an economical method for commercial production of linolenic acid (ω-3 fatty acid).


Assuntos
Ácidos Graxos Insaturados/biossíntese , Fusarium/metabolismo , Biomassa , Cromatografia Gasosa , DNA Intergênico , Ácidos Graxos Ômega-3/análise , Ácidos Graxos Ômega-3/biossíntese , Ácidos Graxos Insaturados/análise , Fusarium/genética , Fusarium/isolamento & purificação , Fusarium/ultraestrutura , Filogenia , RNA Ribossômico 18S/genética , Microbiologia do Solo
15.
Fungal Genet Biol ; 104: 45-55, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28512016

RESUMO

LI-F type peptides (AMP-jsa9) are a group of cyclic lipodepsipeptides that exhibit broad antimicrobial spectrum against Gram-positive bacteria and filamentous fungi. We sought to assess the toxicity of AMP-jsa9 and the mechanism of AMP-jsa9 action against Fusarium moniliforme. AMP-jsa9 exhibited weak hemolytic activity and weak cytotoxicity at antimicrobial concentrations (32µg/ml). Confocal laser microscopy, SEM, and TEM indicated that AMP-jsa9 primarily targets the cell wall, plasma membrane, and cytoskeleton, increases membranepermeability, and enhances cytoplasm leakage (e.g., K+, protein). Quantitative proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ) detected a total of 162 differentially expressed proteins (59 up-regulated and 103 down-regulated) following treatment of F. moniliforme with AMP-jsa9. AMP-jsa9 treatment also led to reductions in chitin, ergosterol, NADH, NADPH, and ATP levels. Moreover, fumonisin B1 expression and biosynthesis was suppressed in AMP-jsa9-treated F. moniliforme. Our results provide a theoretical basis for the application of AMP-jsa9 as a natural and effective antifungal agent in the agricultural, food, and animal feed industries.


Assuntos
Antifúngicos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Fusarium/efeitos dos fármacos , Paenibacillus polymyxa/metabolismo , Adulto , Animais , Antifúngicos/isolamento & purificação , Antifúngicos/toxicidade , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/metabolismo , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/farmacologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Células Cultivadas , Fumonisinas/metabolismo , Proteínas Fúngicas/biossíntese , Fusarium/ultraestrutura , Hemolíticos/farmacologia , Humanos , Camundongos , Testes de Sensibilidade Microbiana , Peptídeos/isolamento & purificação , Proteoma
16.
Sci Rep ; 7: 44365, 2017 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-28295054

RESUMO

Time-lapse microscopic-photography allows in-depth phenotyping of microorganisms. Here we report development of such a system using a microfluidic device, generated from polydimethylsiloxane and glass slide, placed on a motorized stage of a microscope for conducting time-lapse microphotography of multiple observations in 20 channels simultaneously. We have demonstrated the utility of the device in studying growth, germination and sporulation in Fusarium virguliforme that causes sudden death syndrome in soybean. To measure the growth differences, we developed a polyamine oxidase fvpo1 mutant in this fungus that fails to grow in minimal medium containing polyamines as the sole nitrogen source. Using this system, we demonstrated that the conidiospores of the pathogen take an average of five hours to germinate. During sporulation, it takes an average of 10.5 h for a conidiospore to mature and get detached from its conidiophore for the first time. Conidiospores are developed in a single conidiophore one after another. The microfluidic device enabled quantitative time-lapse microphotography reported here should be suitable for screening compounds, peptides, micro-organisms to identify fungitoxic or antimicrobial agents for controlling serious plant pathogens. The device could also be applied in identifying suitable target genes for host-induced gene silencing in pathogens for generating novel disease resistance in crop plants.


Assuntos
Proteínas Fúngicas/genética , Fusarium/ultraestrutura , Dispositivos Lab-On-A-Chip , Micélio/ultraestrutura , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Esporos Fúngicos/ultraestrutura , Dimetilpolisiloxanos/química , Proteínas Fúngicas/metabolismo , Fusarium/genética , Fusarium/crescimento & desenvolvimento , Fusarium/patogenicidade , Expressão Gênica , Vidro/química , Micélio/genética , Micélio/crescimento & desenvolvimento , Micélio/patogenicidade , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Fenótipo , Fotografação/instrumentação , Fotografação/métodos , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Raízes de Plantas/microbiologia , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/patogenicidade , Imagem com Lapso de Tempo/instrumentação , Imagem com Lapso de Tempo/métodos
17.
Mycoses ; 60(3): 204-212, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27928841

RESUMO

Recently, mycetoma was added to the World Health Organization's list of neglected tropical disease priorities. Fusarium as a genus has been reported to cause eumycetoma, but little is known about the species involved in this infection and their identification. In this study, molecular tools were applied to identify Fusarium agents from human eumycetoma cases. The partial translation elongation factor 1-alpha (TEF-1α) gene was used as diagnostic parameter. Two additional cases of eumycetoma, due to F. keratoplasticum and F. pseudensiforme, respectively, are presented. A systematic literature review was performed to assess general features, identification, treatment and outcome of eumycetoma infections due to Fusarium species. Of the 20 reviewed patients, the majority (75%) were male. Most agents belonged to the F. solani species complex, ie F. keratoplasticum, F. pseudensiforme, and an undescribed lineage of F. solani. In addition, F. thapsinum, a member of Fusarium fujikuroi species complex was encountered. The main antifungal drugs used were itraconazole, ketoconazole and amphotericin B, but cure rates were low (15%). Partial response or relapse was observed in some cases, and a case ended in amputation. Clinical management of eumycetoma due to Fusarium is complex and combination therapy might be required to increase cure rates.


Assuntos
Fusariose/tratamento farmacológico , Fusariose/microbiologia , Fusarium/genética , Fusarium/isolamento & purificação , Micetoma/tratamento farmacológico , Micetoma/microbiologia , Adulto , Idoso , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Amputação Cirúrgica , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Feminino , Fusarium/efeitos dos fármacos , Fusarium/ultraestrutura , Humanos , Itraconazol/farmacologia , Itraconazol/uso terapêutico , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Micetoma/diagnóstico , Doenças Negligenciadas/microbiologia , Fator 1 de Elongação de Peptídeos/genética , Filogenia , Recidiva , Análise de Sequência de DNA , Adulto Jovem
18.
Mol Microbiol ; 102(6): 951-974, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27642009

RESUMO

Post-translational modification of histones is a crucial mode of transcriptional regulation in eukaryotes. A well-described acetylation modifier of certain lysine residues is the Spt-Ada-Gcn5 acetyltransferase (SAGA) complex assembled around the histone acetyltransferase Gcn5 in Saccharomyces cerevisiae. We identified and characterized the SAGA complex in the rice pathogen Fusarium fujikuroi, well-known for producing a large variety of secondary metabolites (SMs). By using a co-immunoprecipitation approach, almost all of the S. cerevisiae SAGA complex components have been identified, except for the ubiquitinating DUBm module and the chromodomain containing Chd1. Deletion of GCN5 led to impaired growth, loss of conidiation and alteration of SM biosynthesis. Furthermore, we show that Gcn5 is essential for the acetylation of several histone 3 lysines in F. fujikuroi, that is, H3K4, H3K9, H3K18 and H3K27. A genome-wide microarray analysis revealed differential expression of about 30% of the genome with an enrichment of genes involved in primary and secondary metabolism, transport and histone modification. HPLC-based analysis of known SMs revealed significant alterations in the Δgcn5 mutant. While most SM genes were activated by Gcn5 activity, the biosynthesis of the pigment bikaverin was strongly increased upon GCN5 deletion underlining the diverse roles of the SAGA complex in F. fujikuroi.


Assuntos
Acetiltransferases/metabolismo , Fusarium/metabolismo , Acetilação , Acetiltransferases/genética , Proteínas de Ligação a DNA/metabolismo , Fusarium/ultraestrutura , Histonas/metabolismo , Imunoprecipitação , Oryza/microbiologia , Processamento de Proteína Pós-Traducional , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Relação Estrutura-Atividade , Transativadores/metabolismo , Fatores de Transcrição/metabolismo
19.
Rev Argent Microbiol ; 48(3): 222-228, 2016.
Artigo em Espanhol | MEDLINE | ID: mdl-27590608

RESUMO

Garlic in Mexico is one of the most profitable vegetable crops, grown in almost 5,451ha; out of which more than 83% are located in Zacatecas, Guanajuato, Sonora, Puebla, Baja California and Aguascalientes. Blossom-end rot caused by Fusarium spp is widely distributed worldwide and has been a limiting factor in onion and garlic production regions, not only in Mexico but also in other countries. The presence of Fusarium oxysporum has been reported in Guanajuato and Aguascalientes. Fusarium culmorum has been reported in onion cultivars of Morelos; and Fusarium proliferatum, Fusarium verticillioides, Fusarium solani and Fusarium acuminatum have been previously reported in Aguascalientes. The goal of this work was identifying the Fusarium species found in Zacatecas, Guanajuato and Aguascalientes, to assess their pathogenicity. Plants with disease symptoms were collected from hereinabove mentioned States. The samples resulted in the identification of: F. oxysporum, F. proliferatum, F. verticillioides, F. solani and F. acuminatum species; out of which Aguascalientes AGS1A (F. oxysporum), AGS1B (F. oxysporum) and AGSY-10 (F. acuminatum) strains showed higher severity under greenhouse conditions.


Assuntos
Produtos Agrícolas/microbiologia , Fusarium/isolamento & purificação , Alho/microbiologia , Doenças das Plantas/microbiologia , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Fusarium/classificação , Fusarium/patogenicidade , Fusarium/ultraestrutura , México , Especificidade da Espécie , Virulência
20.
Langmuir ; 32(34): 8688-97, 2016 08 30.
Artigo em Inglês | MEDLINE | ID: mdl-27466012

RESUMO

Understanding the synthesis and properties of nanomaterials is critical for reliable applications. Biological systems, such as fungi, have been described as a "green" alternative to synthesis, yet knowledge gaps exist in terms of production variability, comparison with commercial products, and identifying a clear biological advantage over other synthesis methods. In this study, we evaluated 12 fungal isolates of Fusarium oxysporum for Ag nanoparticle production and characterized the resultant biologically produced (biogenic) nanoparticles. We show evidence that isolate selection, temperature, and pH can influence the quantity, size, and shape of nanoparticles. All F. oxysporum isolates produced Ag nanoparticles, but in varied quantities. Increasing incubation temperature increased the quantity, yet nanoparticle diameter was inversely related to temperature. Variations in pH predominately influenced nanoparticle morphology. A direct comparison with commercial, chemically produced Ag nanoparticles yielded physical similarities; however, important differences in surface chemistry are observed. Biogenic nanoparticles show a greater degree of colloidal stability in high-ionic-strength solutions, pointing to a biological advantage associated with the fungal produced layer (corona) surrounding the nanoparticles. It is clear that understanding the organic layer and interfacial interactions will be beneficial in developing innovative applications, particularly in the field of biosensing.


Assuntos
Fusarium/isolamento & purificação , Fusarium/metabolismo , Nanopartículas Metálicas/química , Coloides , Estabilidade de Medicamentos , Difusão Dinâmica da Luz , Fusarium/ultraestrutura , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Nanotecnologia , Tamanho da Partícula , Prata/química , Propriedades de Superfície , Temperatura
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