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1.
Mol Divers ; 23(2): 299-306, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30168050

RESUMO

A series of novel Mannich base derivatives of flavone containing benzylamine moiety was synthesized using the Mannich reaction. The results of antifungal activity are not ideal, but its antifungal effect has a certain increase compared to flavonoids. After that, four bacteria were used to test antibacterial experiments of these compounds; compound 5g (MIC = 0.5, 0.125 mg/L) showed significant inhibitory activity against Staphylococcus aureus and Salmonella gallinarum compared with novobiocin (MIC = 2, 0.25 mg/L). Compound 5s exhibited broad spectrum antibacterial activity (MIC = 1, 0.5, 2, 0.05 mg/L) against four bacteria. The selected compounds 5g and 5s exhibit potent inhibition against Topo II and Topo IV with IC50 values (0.25-16 mg/L). Molecular docking model showed that the compounds 5g and 5s can bind well to the target by interacting with amino acid residues. It will provide some valuable information for the commercial antibacterial agents.


Assuntos
Antibacterianos , DNA Topoisomerase IV/antagonistas & inibidores , Flavonas , Bases de Mannich , Inibidores da Topoisomerase II , Antibacterianos/química , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Desenho de Fármacos , Flavonas/química , Flavonas/farmacologia , Gibberella/efeitos dos fármacos , Gibberella/crescimento & desenvolvimento , Bases de Mannich/química , Bases de Mannich/farmacologia , Simulação de Acoplamento Molecular , Relação Estrutura-Atividade , Inibidores da Topoisomerase II/química , Inibidores da Topoisomerase II/farmacologia
2.
World J Microbiol Biotechnol ; 32(11): 182, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27646209

RESUMO

This study determined the specific uptake rate of glucose and corn oil substrates used as carbon sources in batch cultures of Gibberella fujikuroi. We tested three biological models of growth rate: Monod, logistic and lag-exponential. With respect to the substrate consumption rate, we tested two models: constant cell yield (CCY) and law of mass action (LMA). The experimental data obtained from the culture with glucose as substrate correlated satisfactorily with the logistic/LMA model, indicating that the cell yield was variable. In the case of corn oil as carbon source, considering total residual lipids as substrate in the culture broth, the model with the best correlation was the lag-exp/CCY model. The quantification by GC of the three main fatty acids (linoleic, oleic and palmitic) in the culture medium showed a cumulative behavior, with a maximum concentration of each acid at 36 h. We established a more explicit mechanism of the consumption of corn oil, consisting of two stages: generation of fatty acids by hydrolysis and consumption by cellular uptake. The kinetic of hydrolysable lipids was of first order. We found that the hydrolysis rate of corn oil is not a limiting factor for the uptake of fatty acids by the microorganism. We also established, based on the analysis of the identical mathematical structure of consumption kinetics, that the uptake of fatty acids is faster than the uptake of glucose.


Assuntos
Técnicas de Cultura Celular por Lotes/métodos , Óleo de Milho/metabolismo , Gibberella/crescimento & desenvolvimento , Glucose/metabolismo , Biomassa , Carbono/metabolismo , Meios de Cultura , Cinética , Lipídeos/química , Modelos Logísticos
3.
Prep Biochem Biotechnol ; 44(1): 80-9, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24117154

RESUMO

Gibberelic acid fermentation using extractive methods was carried out in the presence of corn oil and Alamine 336. Gibberella fujikuroi fungus (NRRL 2278) was used to produce gibberellic acid. Oleyl alcohol was a diluting agent for Alamine 336. The effects of oleyl alcohol (100%, v/v), corn oil (5-25%, v/v), the concentration of Alamine 336 in oleyl alcohol, and feeding air were examined in this study. According to the results, oleyl alcohol was not effective on the production. On the other hand, oleyl alcohol solutions containing 15-30% (v/v) Alamine 336 showed effects as a toxic substance. In order to reduce solvent toxicity, corn oil was used. Addition of corn oil increased the concentration of gibberellic acid 1.3-fold compared to the control. Then the effects of immobilization and co-immobilization on extractive gibberelic acid fermentation were investigated. The highest total gibberellic acid concentration of 158.9 mg/L was produced with immobilized cells and feeding air by using extractive fermentation. The yield of gibberellic acid increased about 2.6-fold compared with the shake-flask fermentation (60.5 mg/L) without organic solutions.


Assuntos
Óleo de Milho/farmacologia , Álcoois Graxos/farmacologia , Gibberella/crescimento & desenvolvimento , Giberelinas/biossíntese , Células Imobilizadas/metabolismo , Gibberella/química , Gibberella/metabolismo , Giberelinas/química , Giberelinas/isolamento & purificação
4.
Eukaryot Cell ; 11(9): 1143-53, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22798392

RESUMO

Fungi have evolved efficient metabolic mechanisms for the exact temporal (developmental stages) and spatial (organelles) production of acetyl coenzyme A (acetyl-CoA). We previously demonstrated mechanistic roles of several acetyl-CoA synthetic enzymes, namely, ATP citrate lyase and acetyl-CoA synthetases (ACSs), in the plant-pathogenic fungus Gibberella zeae. In this study, we characterized two carnitine acetyltransferases (CATs; CAT1 and CAT2) to obtain a better understanding of the metabolic processes occurring in G. zeae. We found that CAT1 functioned as an alternative source of acetyl-CoA required for lipid accumulation in an ACS1 deletion mutant. Moreover, deletion of CAT1 and/or CAT2 resulted in various defects, including changes to vegetative growth, asexual/sexual development, trichothecene production, and virulence. Although CAT1 is associated primarily with peroxisomal CAT function, mislocalization experiments showed that the role of CAT1 in acetyl-CoA transport between the mitochondria and cytosol is important for sexual and asexual development in G. zeae. Taking these data together, we concluded that G. zeae CATs are responsible for facilitating the exchange of acetyl-CoA across intracellular membranes, particularly between the mitochondria and the cytosol, during various developmental stages.


Assuntos
Acetilcoenzima A/metabolismo , Carnitina Aciltransferases/metabolismo , Proteínas Fúngicas/metabolismo , Gibberella/crescimento & desenvolvimento , Gibberella/metabolismo , Acetato-CoA Ligase/genética , Acetato-CoA Ligase/metabolismo , Transporte Biológico , Carnitina Aciltransferases/genética , Citosol/metabolismo , Proteínas Fúngicas/genética , Deleção de Genes , Gibberella/genética , Gibberella/patogenicidade , Mitocôndrias/metabolismo , Peroxissomos/metabolismo , Reprodução Assexuada , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , Tricotecenos/biossíntese , Tricotecenos/genética , Virulência/genética
5.
Eukaryot Cell ; 10(6): 832-41, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21357477

RESUMO

The role of Mid1, a stretch-activated ion channel capable of being permeated by calcium, in ascospore development and forcible discharge from asci was examined in the pathogenic fungus Gibberella zeae (anamorph Fusarium graminearum). The Δmid1 mutants exhibited a >12-fold reduction in ascospore discharge activity and produced predominately abnormal two-celled ascospores with constricted and fragile septae. The vegetative growth rate of the mutants was ∼50% of the wild-type rate, and production of macroconidia was >10-fold lower than in the wild type. To better understand the role of calcium flux, Δmid1 Δcch1 double mutants were also examined, as Cch1, an L-type calcium ion channel, is associated with Mid1 in Saccharomyces cerevisiae. The phenotype of the Δmid1 Δcch1 double mutants was similar to but more severe than the phenotype of the Δmid1 mutants for all categories. Potential and current-voltage measurements were taken in the vegetative hyphae of the Δmid1 and Δcch1 mutants and the wild type, and the measurements for all three strains were remarkably similar, indicating that neither protein contributes significantly to the overall electrical properties of the plasma membrane. Pathogenicity of the Δmid1 and Δmid1Δcch1 mutants on the host (wheat) was not affected by the mutations. Exogenous calcium supplementation partially restored the ascospore discharge and vegetative growth defects for all mutants, but abnormal ascospores were still produced. These results extend the known roles of Mid1 to ascospore development and forcible discharge. However, Neurospora crassa Δmid1 mutants were also examined and did not exhibit defects in ascospore development or in ascospore discharge. In comparison to ion channels in other ascomycetes, Mid1 shows remarkable adaptability of roles, particularly with regard to niche-specific adaptation.


Assuntos
Canais de Cálcio/metabolismo , Proteínas Fúngicas/metabolismo , Gibberella/crescimento & desenvolvimento , Mecanotransdução Celular , Esporos Fúngicos/metabolismo , Sinalização do Cálcio/genética , Proliferação de Células , Meios de Cultura , Técnicas de Inativação de Genes , Gibberella/fisiologia , Potenciais da Membrana
6.
Eukaryot Cell ; 10(8): 1043-52, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21666077

RESUMO

Acetyl coenzyme A (acetyl-CoA) is a crucial metabolite for energy metabolism and biosynthetic pathways and is produced in various cellular compartments with spatial and temporal precision. Our previous study on ATP citrate lyase (ACL) in Gibberella zeae revealed that ACL-dependent acetyl-CoA production is important for histone acetylation, especially in sexual development, but is not involved in lipid synthesis. In this study, we deleted additional acetyl-CoA synthetic genes, the acetyl-CoA synthetases (ACS genes ACS1 and ACS2), to identify alternative acetyl-CoA production mechanisms for ACL. The ACS1 deletion resulted in a defect in sexual development that was mainly due to a reduction in 1-palmitoyl-2-oleoyl-3-linoleoyl-rac-glycerol production, which is required for perithecium development and maturation. Another ACS coding gene, ACS2, has accessorial functions for ACS1 and has compensatory functions for ACL as a nuclear acetyl-CoA producer. This study showed that acetate is readily generated during the entire life cycle of G. zeae and has a pivotal role in fungal metabolism. Because ACSs are components of the pyruvate-acetaldehyde-acetate pathway, this fermentation process might have crucial roles in various physiological processes for filamentous fungi.


Assuntos
Acetato-CoA Ligase/metabolismo , Proteínas Fúngicas/metabolismo , Gibberella/enzimologia , Acetato-CoA Ligase/genética , Acetatos/metabolismo , Acetilcoenzima A/biossíntese , Proteínas Fúngicas/genética , Deleção de Genes , Expressão Gênica , Engenharia Genética , Gibberella/genética , Gibberella/crescimento & desenvolvimento , Metabolismo dos Lipídeos , Micélio/genética , Micélio/crescimento & desenvolvimento , Micélio/metabolismo , Regiões Promotoras Genéticas , Triglicerídeos/metabolismo
7.
Curr Microbiol ; 64(5): 469-76, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22350001

RESUMO

To determine the feasibility of inducing mutation for Gibberella moniliformis EZG0807 with a superconducting magnet, this paper investigated the effects of this instrument on the filamentous fungus G. moniliformis EZG0807. The superconducting magnet could simulate space gravity environment from hypo-gravity (0 g) to hyper-gravity (2 g). After G. moniliformis EZG0807 was exposed to the superconducting magnet for 72 h, the morphological observation, agar diffusion method, and amplified fragment length polymorphism were performed to detect the mutagenic effects in the aspect of morphology, the activity of metabolites, and genomic DNA, respectively. The mutant strain M7212 in 1 g (16 T) was different from the control in the morphology, showing no activity against the four tested bacteria Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Proteus vulgaris, and lost a size of 675 bp band on the genomic DNA. These results indicated that the superconducting magnet could be used to induce mutation for G. moniliformis EZG0807, which enabled improving the production of G. moniliformis EZG0807 and providing an effective approach for fungal breeding.


Assuntos
Técnicas Genéticas/instrumentação , Gibberella/genética , Mutagênese , Bacillus subtilis/química , Bacillus subtilis/genética , Escherichia coli/química , Escherichia coli/genética , Gibberella/química , Gibberella/crescimento & desenvolvimento , Gravitação , Imãs , Mutação , Staphylococcus aureus/química , Staphylococcus aureus/genética
8.
Molecules ; 17(5): 5139-50, 2012 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-22555301

RESUMO

Sixteen novel pyrazole acyl thiourea derivatives 6 were synthesized from monomethylhydrazine (phenylhydrazine) and ethyl acetoacetate. The key 5-chloro-3-methyl-1-substituted-1H-pyrazole-4-carbonyl chloride intermediates 4 were first generated in four steps through cyclization, formylation, oxidation and acylation. Thess were then reacted with ammonium thiocyanate in the presence of PEG-400 to afford 5-chloro-3-methyl-1-substituted-1H-pyrazole-4-carbonyl isothiocyanates 5. Subsequent reaction with fluorinated aromatic amines resulted in the formation of the title compounds. The synthesized compound were unequivocally characterized by IR, ¹H-NMR, ¹³C-NMR and elemental analysis and some of the synthesized compounds displayed good antifungal activities against Gibberella zeae, Fusarium oxysporum, Cytospora mandshurica and anti-TMV activity in preliminary antifungal activity tests.


Assuntos
Antifúngicos/síntese química , Antivirais/síntese química , Fusarium/efeitos dos fármacos , Gibberella/efeitos dos fármacos , Pirazóis/síntese química , Tioureia/síntese química , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Acetoacetatos/química , Acilação , Antifúngicos/farmacologia , Antivirais/farmacologia , Ciclização , Fusarium/crescimento & desenvolvimento , Gibberella/crescimento & desenvolvimento , Hidrocarbonetos Fluorados/química , Testes de Sensibilidade Microbiana , Oxirredução , Fenil-Hidrazinas/química , Polietilenoglicóis/química , Pirazóis/farmacologia , Relação Estrutura-Atividade , Tiocianatos/química , Tioureia/análogos & derivados , Tioureia/farmacologia , Vírus do Mosaico do Tabaco/fisiologia
9.
Fungal Genet Biol ; 48(4): 408-17, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21237280

RESUMO

Adenosine triphosphate (ATP) citrate lyase (ACL) is a key enzyme in the production of cytosolic acetyl-CoA, which is crucial for de novo lipid synthesis and histone acetylation in mammalian cells. In this study, we characterized the mechanistic roles of ACL in the homothallic ascomycete fungus Gibberella zeae, which causes Fusarium head blight in major cereal crops. Deletion of ACL in the fungus resulted in a complete loss of self and female fertility as well as a reduction in asexual reproduction, virulence, and trichothecene production. When the wild-type strain was spermatized with the ACL deletion mutants, they produced viable ascospores, however ascospore delimitation was not properly regulated. Although lipid synthesis was not affected by ACL deletion, histone acetylation was dramatically reduced in the ACL deletion mutants during sexual development, suggesting that the defects in sexual reproduction were caused by the reduction in histone acetylation. This study is the first report demonstrating a link between sexual development and ACL-mediated histone acetylation in fungi.


Assuntos
ATP Citrato (pro-S)-Liase/metabolismo , Gibberella/enzimologia , Gibberella/crescimento & desenvolvimento , ATP Citrato (pro-S)-Liase/genética , Acetilação , Deleção de Genes , Genes Fúngicos , Gibberella/genética , Histonas/metabolismo , Esporos Fúngicos/crescimento & desenvolvimento , Tricotecenos/metabolismo
10.
Eukaryot Cell ; 9(10): 1495-503, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20802018

RESUMO

Head blight, caused by Gibberella zeae, is a significant disease among cereal crops, including wheat, barley, and rice, due to contamination of grain with mycotoxins. G. zeae is spread by ascospores forcibly discharged from sexual fruiting bodies forming on crop residues. In this study, we characterized a novel gene, ROA, which is required for normal sexual development. Deletion of ROA (Δroa) resulted in an abnormal size and shape of asci and ascospores but did not affect vegetative growth. The Δroa mutation triggered round ascospores and insufficient cell division after spore delimitation. The asci of the Δroa strain discharged fewer ascospores from the perithecia but achieved a greater dispersal distance than those of the wild-type strain. Turgor pressure within the asci was calculated through the analysis of osmolytes in the epiplasmic fluid. Deletion of the ROA gene appeared to increase turgor pressure in the mutant asci. The higher turgor pressure of the Δroa mutant asci and the mutant spore shape contributed to the longer distance dispersal. When the Δroa mutant was outcrossed with a Δmat1-2 mutant, a strain that contains a green fluorescence protein (GFP) marker in place of the MAT1-2 gene, unusual phenotypic segregation occurred. The ratio of GFP to non-GFP segregation was 1:1; however, all eight spores had the same shape. Taken together, the results of this study suggest that ROA plays multiple roles in maintaining the proper morphology and discharge of ascospores in G. zeae.


Assuntos
Grão Comestível/microbiologia , Genes Fúngicos , Gibberella/crescimento & desenvolvimento , Gibberella/fisiologia , Doenças das Plantas/microbiologia , Esporos Fúngicos/fisiologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Gibberella/genética , Gibberella/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Morfogênese , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/metabolismo
11.
Appl Environ Microbiol ; 76(10): 3089-96, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20348311

RESUMO

The ascomycete fungus Gibberella zeae is an important plant pathogen that causes fusarium head blight on small grains. Molecular studies of this fungus have been performed extensively to uncover the biological mechanisms related to pathogenicity, toxin production, and sexual reproduction. Molecular methods, such as targeted gene deletion, gene overexpression, and gene fusion to green fluorescent protein (GFP), are relatively easy to perform with this fungus; however, conditional expression systems have not been developed. The purpose of this study was to identify a promoter that could be induced by zearalenone (ZEA) for the development of a conditional expression system in G. zeae. Through microarray analysis, we isolated one zearalenone response gene (ZEAR) whose expression was increased more than 50 times after ZEA treatment. Northern blot analysis showed that the ZEAR transcript dramatically increased after 1 h of ZEA treatment. To determine the utility of the ZEAR promoter, called Pzear, in a conditional expression system, we transformed a Pzear::GFP fusion construct into G. zeae. Our data showed a ZEA concentration-dependent increase in GFP expression. We also replaced the promoter of G. zeae metE (GzmetE), an essential gene for methionine biosynthesis, with the Pzear promoter. The growth of the Pzear-GzmetE mutant on minimal medium was dependent on the ZEA concentration supplemented in the medium and showed that GzMetE expression was induced by ZEA. This study is the first report of an inducible promoter in G. zeae. Our system will be useful for the characterization of essential gene functions in this fungus through differential and ZEA-dependent gene expression. In addition, the Pzear promoter may be applicable as a biosensor for the detection of ZEA contamination in agricultural products.


Assuntos
Perfilação da Expressão Gênica/métodos , Regulação Fúngica da Expressão Gênica , Gibberella/genética , Gibberella/metabolismo , Regiões Promotoras Genéticas/genética , Zearalenona/metabolismo , Deleção de Genes , Gibberella/crescimento & desenvolvimento , Proteínas de Fluorescência Verde/genética , Regiões Promotoras Genéticas/efeitos dos fármacos , Sequências Reguladoras de Ácido Nucleico/genética , Zearalenona/farmacologia
12.
Eukaryot Cell ; 8(1): 116-27, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19028993

RESUMO

The sucrose nonfermenting 1 (SNF1) protein kinase of yeast plays a central role in the transcription of glucose-repressible genes in response to glucose starvation. In this study, we deleted an ortholog of SNF1 from Gibberella zeae to characterize its functions by using a gene replacement strategy. The mycelial growth of deletion mutants (DeltaGzSNF1) was reduced by 21 to 74% on diverse carbon sources. The virulence of DeltaGzSNF1 mutants on barley decreased, and the expression of genes encoding cell-wall-degrading enzymes was reduced. The most distinct phenotypic changes were in sexual and asexual development. DeltaGzSNF1 mutants produced 30% fewer perithecia, which matured more slowly, and asci that contained one to eight abnormally shaped ascospores. Mutants in which only the GzSNF1 catalytic domain was deleted had the same phenotype changes as the DeltaGzSNF1 strains, but the phenotype was less extreme in the mutants with the regulatory domain deleted. In outcrosses between the DeltaGzSNF1 mutants, each perithecium contained approximately 70% of the abnormal ascospores, and approximately 50% of the asci showed unexpected segregation patterns in a single locus tested. The asexual spores of the DeltaGzSNF1 mutants were shorter and had fewer septa than those of the wild-type strain. The germination and nucleation of both ascospores and conidia were delayed in DeltaGzSNF1 mutants in comparison with those of the wild-type strain. GzSNF1 expression and localization depended on the developmental stage of the fungus. These results suggest that GzSNF1 is critical for normal sexual and asexual development in addition to virulence and the utilization of alternative carbon sources.


Assuntos
Proteínas Fúngicas/metabolismo , Gibberella/enzimologia , Gibberella/crescimento & desenvolvimento , Hordeum/microbiologia , Doenças das Plantas/microbiologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Regulação da Expressão Gênica no Desenvolvimento , Regulação Fúngica da Expressão Gênica , Genes Fúngicos Tipo Acasalamento , Gibberella/genética , Gibberella/patogenicidade , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/genética , Estrutura Terciária de Proteína , Esporos Fúngicos/enzimologia , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/patogenicidade , Virulência
13.
Eukaryot Cell ; 8(8): 1155-64, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19525419

RESUMO

The glyoxylate and methylcitrate cycles are involved in the metabolism of two- or three-carbon compounds in fungi. To elucidate the role(s) of these pathways in Gibberella zeae, which causes head blight in cereal crops, we focused on the functions of G. zeae orthologs (GzICL1 and GzMCL1) of the genes that encode isocitrate lyase (ICL) and methylisocitrate lyase (MCL), respectively, key enzymes in each cycle. The deletion of GzICL1 (DeltaGzICL1) caused defects in growth on acetate and in perithecium (sexual fruiting body) formation but not in virulence on barley and wheat, indicating that GzICL1 acts as the ICL of the glyoxylate cycle and is essential for self-fertility in G. zeae. In contrast, the DeltaGzMCL1 strains failed to grow on propionate but exhibited no major changes in other traits, suggesting that GzMCL1 is required for the methylcitrate cycle in G. zeae. Interestingly, double deletion of both GzICL1 and GzMCL1 caused significantly reduced virulence on host plants, indicating that both GzICL1 and GzMCL1 have redundant functions for plant infection in G. zeae. Thus, both GzICL1 and GzMCL1 may play important roles in determining major mycological and pathological traits of G. zeae by participating in different metabolic pathways for the use of fatty acids.


Assuntos
Citratos/metabolismo , Grão Comestível/microbiologia , Gibberella/crescimento & desenvolvimento , Gibberella/patogenicidade , Glioxilatos/metabolismo , Doenças das Plantas/microbiologia , Carbono-Carbono Liases/genética , Carbono-Carbono Liases/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Gibberella/enzimologia , Gibberella/metabolismo , Isocitrato Liase/genética , Isocitrato Liase/metabolismo , Virulência
14.
Eukaryot Cell ; 7(2): 415-24, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18083828

RESUMO

Cch1, a putative voltage-gated calcium ion channel, was investigated for its role in ascus development in Gibberella zeae. Gene replacement mutants of CCH1 were generated and found to have asci which did not forcibly discharge spores, although morphologically ascus and ascospore development in the majority of asci appeared normal. Additionally, mycelial growth was significantly slower, and sexual development was slightly delayed in the mutant; mutant mycelia showed a distinctive fluffy morphology, and no cirrhi were produced. Wheat infected with Deltacch1 mutants developed symptoms comparable to wheat infected with the wild type; however, the mutants showed a reduced ability to protect the infected stalk from colonization by saprobic fungi. Transcriptional analysis of gene expression in mutants using the Affymetrix Fusarium microarray showed 2,449 genes with significant, twofold or greater, changes in transcript abundance across a developmental series. This work extends the role of CCH1 to forcible spore discharge in G. zeae and suggests that this channel has subtle effects on growth and development.


Assuntos
Canais de Cálcio Tipo L/metabolismo , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Gibberella/genética , Esporos Fúngicos/fisiologia , Triticum/microbiologia , Southern Blotting , Cálcio/metabolismo , Canais de Cálcio Tipo L/genética , Cryptococcus neoformans/genética , Cryptococcus neoformans/crescimento & desenvolvimento , Proteínas Fúngicas/metabolismo , Perfilação da Expressão Gênica , Gibberella/crescimento & desenvolvimento , Análise de Sequência com Séries de Oligonucleotídeos , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , RNA Fúngico/genética , RNA Fúngico/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Triticum/genética
15.
Int J Food Microbiol ; 290: 262-272, 2019 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-30408647

RESUMO

Contamination with filamentous fungi during cocoa bean fermentation and drying reduces the quality of cocoa beans and poses a health risk for consumers due to the potential accumulation of mycotoxins. The aim of this study was to develop anti-fungal lactic acid bacteria (LAB)-yeast co-cultures by selecting anti-fungal strains best adapted to the cocoa bean fermentation process from 362 LAB and 384 yeast strains isolated from cocoa bean post-harvest processes. The applied multiphasic screening approach included anti-fungal activity tests in vitro and in vivo and assessment of the carbon metabolism and stress tolerance of the anti-fungal strains in a cocoa pulp simulation medium. The anti-fungal strains, Lactobacillus fermentum M017, Lb. fermentum 223, Hanseniaspora opuntiae H17, and Saccharomyces cerevisiae H290, were selected based on their high fungal growth inhibition capacity and their well-adapted metabolism. Up to seven filamentous fungal strains of the genera Aspergillus, Penicillium, and Gibberella were inhibited on average by 63 and 75% of the maximal inhibition zone by M017 and 223, respectively, and by 25 and 31% by the strains H17 and H290, respectively. Both Lb. fermentum strains converted the medium's glucose, fructose, and citric acid into 20.4-23.0 g/l of mannitol, 3.9-6.2 g/l acetic acid, and 8.6-10.3 g/l lactic acid, whereas the two yeast strains metabolized glucose and fructose to produce 7.4-18.4 g/l of ethanol. The Lb. fermentum strains were further characterized as particularly tolerant towards ethanol, acetic acid, and heat stress and both yeast strains tolerated high amounts of ethanol and lactic acid in the medium. Finally, the anti-fungal in vivo assays revealed that the two Lb. fermentum strains completely inhibited growth of the citrinin-producing strain, P. citrinum S005, and the potentially fumonisin-producing strain, G. moniliformis S003, on the surface of cocoa beans. Furthermore, growth of the aflatoxin-producer A. flavus S075 was inhibited after 10-14 days by all four selected anti-fungal strains, i.e. Lb. fermentum M017, Lb. fermentum 223, H. opuntiae H17, and Sacc. cerevisiae H290, at 51-95% when applied as single cultures and at 100% when the strains were combined into four co-cultures, each composed of a Lb. fermentum and one of the two yeast strains. As a conclusion, these four LAB-yeast co-cultures are recommended for future applications to limit the growth of filamentous fungi and the concomitant mycotoxin production during the fermentation of cocoa beans.


Assuntos
Cacau/microbiologia , Fermentação , Lactobacillales/metabolismo , Saccharomyces cerevisiae/metabolismo , Ácido Acético/metabolismo , Aflatoxinas/análise , Aspergillus flavus/crescimento & desenvolvimento , Agentes de Controle Biológico/metabolismo , Técnicas de Cocultura , Etanol/metabolismo , Contaminação de Alimentos/prevenção & controle , Microbiologia de Alimentos , Gibberella/crescimento & desenvolvimento , Hanseniaspora/metabolismo , Resposta ao Choque Térmico , Ácido Láctico/metabolismo , Limosilactobacillus fermentum/metabolismo , Penicillium/crescimento & desenvolvimento
16.
Phytopathology ; 98(5): 504-8, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18943217

RESUMO

Fusarium head blight (FHB) is one of the most important cereal diseases in the world and has caused major losses to the grain industry. The principal pathogen causing FHB in North America is Gibberella zeae (anamorph Fusarium graminearum). Information on survival and the conditions under which ascospores remain viable once released from perithecia may assist in refining disease forecasting models. This study measured germination of ascospores after exposure to different temperatures, 15, 20, and 30 degrees C, and levels of relative humidity (RH), 30, 60, and 90% for 4, 24, or 48 h periods. Viability was tested by germination on water agar. Germination rates fell with increasing temperatures at all observation times and at all humidity levels. At 15 and 20 degrees C after 48 h, germination ranged from 74 to 85%, and 52 to 72%, respectively. At 30 degrees C, germination ranged from 36 to 59% after 24 h and from 13 to 47% after 48 h. Germination was highest at 90% RH, except at 30 degrees C after 48 h, and lowest at 60% RH. Successful germination, even under extreme conditions, suggests that ascospores are sufficiently robust to constitute a source of inoculum under most environmental conditions encountered during the growing season.


Assuntos
Gibberella/crescimento & desenvolvimento , Umidade , Esporos Fúngicos/crescimento & desenvolvimento , Temperatura , Fatores de Tempo
17.
Environ Sci Pollut Res Int ; 25(25): 24965-24974, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29931648

RESUMO

In this paper, a highly copper-resistant fungal strain NT-1 was characterized by morphological, physiological, biochemical, and molecular biological techniques. Physiological response to Cu(II) stress, effects of environmental factors on Cu(II) biosorption, as well as mechanisms of Cu(II) biosorption by strain NT-1 were also investigated in this study. The results showed that NT-1 belonged to the genus Gibberella, which exhibited high tolerance to both acidic conditions and Cu(II) contamination in the environment. High concentrations of copper stress inhibited the growth of NT-1 to various degrees, leading to the decreases in mycelial biomass and colony diameter, as well as changes in morphology. Under optimal conditions (initial copper concentration: 200 mg L-1, temperature 28 °C, pH 5.0, and inoculum dose 10%), the maximum copper removal percentage from solution through culture of strain NT-1 within 5 days reached up to 45.5%. The biosorption of Cu(II) by NT-1 conformed to quasi-second-order kinetics and Langmuir isothermal adsorption model and was confirmed to be a monolayer adsorption process dominated by surface adsorption. The binding of NT-1 to Cu(II) was mainly achieved by forming polydentate complexes with carboxylate and amide group through covalent interactions and forming Cu-nitrogen-containing heterocyclic complexes via Cu(II)-π interaction. The results of this study provide a new fungal resource and key parameters influencing growth and copper removal capacity of the strain for developing an effective bioremediation strategy for copper-contaminated acidic orchard soils.


Assuntos
Cobre/análise , Gibberella/crescimento & desenvolvimento , Poluentes do Solo/análise , Solo/química , Adsorção , Biodegradação Ambiental , Biomassa , Cobre/toxicidade , Gibberella/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Cinética , Poluentes do Solo/toxicidade , Temperatura
18.
Int J Biol Macromol ; 106: 68-77, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28774807

RESUMO

A novel type of O-carboxymethyl chitosan Schiff bases (O-CSPX) was synthesized via a condensation reaction. After the coordination reaction of cupric ions, zinc ions and nickel ions, metal complexes (O-CSPX-M) were achieved. The theoretical structure of O-CSPX-M calculated by Gaussian 09 reveals that the copper ions and nickel ions underwent dsp2 hybridization, the zinc ions underwent sp3 hybridization, and they all coordinated by the carbon atom in the p-π conjugate group. Then, the structures were confirmed by FT-IR, 1H NMR, CP-MAS 13C NMR, elemental analysis, DSC and XRD. The antifungal properties of O-CSPX-M against Phytophthora capsici (P. capsici), Gibberella zeae (G. zeae), Fusarium oxysporum (F. oxysporum) and Botrytis cinerea (B. cinerea) were evaluated at concentrations ranging from 0.05mg/mL to 0.40mg/mL. The experiments indicated that the derivatives have significantly enhanced antifungal activity after metal ions complexation compared with the original chitosan. Moreover, it was shown that 0.20mg/mL of O-CSPX-Cu can 100% inhibit the growth of P. capsici and 0.20mg/mL of O-CSPX-Ni can 87.5% inhibit the growth of B. cinerea. In addition, the phytotoxicity assay and cell viability assay were also evaluated. The experimental results may provide a novel direction for the development of metal fungicides.


Assuntos
Antifúngicos/síntese química , Quitosana/análogos & derivados , Complexos de Coordenação/síntese química , Cobre/química , Níquel/química , Zinco/química , Animais , Antifúngicos/farmacologia , Botrytis/efeitos dos fármacos , Botrytis/crescimento & desenvolvimento , Cátions Bivalentes , Sobrevivência Celular/efeitos dos fármacos , Quitosana/síntese química , Quitosana/farmacologia , Complexos de Coordenação/farmacologia , Cobre/farmacologia , Fusarium/efeitos dos fármacos , Fusarium/crescimento & desenvolvimento , Germinação/efeitos dos fármacos , Gibberella/efeitos dos fármacos , Gibberella/crescimento & desenvolvimento , Camundongos , Testes de Sensibilidade Microbiana , Níquel/farmacologia , Phytophthora/efeitos dos fármacos , Phytophthora/crescimento & desenvolvimento , Células RAW 264.7 , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Triticum/efeitos dos fármacos , Triticum/fisiologia , Zinco/farmacologia
19.
Int J Food Microbiol ; 119(1-2): 126-30, 2007 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-17716767

RESUMO

Many mycotoxigenic fungi infect plant hosts and cause disease in the field. Therefore, control of field infection by these fungi is a critical step in managing mycotoxin accumulation in the harvested product. Fusarium graminearum, also known as Gibberella zeae, is the causal agent of Fusarium head blight (FHB), or scab, in cereals and is also the primary agent responsible for contamination of grain with deoxynivalenol (DON). Research efforts worldwide are devoted to the development of strategies to control field infection of wheat and barley by this pathogen. Strategies include the use of fungicides and biological control agents to protect flowering heads from infection. There is extensive effort in breeding for host resistance to infection and spread of the pathogen within the heads. Scientists are also seeking exogenous traits to introduce into cereals to enhance resistance. Cultural practices are also being examined, primarily as measures to reduce pathogen survival and inoculum production in crop residues. The successes and limitations of these strategies in the management of Fusarium head blight and deoxynivalenol are discussed.


Assuntos
Contaminação de Alimentos/prevenção & controle , Gibberella/crescimento & desenvolvimento , Tricotecenos/análise , Triticum/química , Triticum/microbiologia , Contaminação de Alimentos/análise , Fungicidas Industriais/farmacologia , Gibberella/metabolismo , Controle Biológico de Vetores , Doenças das Plantas/microbiologia , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/microbiologia , Gestão de Riscos
20.
Pest Manag Sci ; 73(3): 546-553, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27158946

RESUMO

BACKGROUND: A 2 year study was conducted to determine whether western bean cutworm (Striacosta albicosta Smith) (WBC) larval feeding damage increases severity of the fungal disease Gibberella ear rot [Fusarium graminearum (Schwein.) Petch] in field corn (Zea mays L.). The effect of a quinone-outside inhibiting fungicide, pyraclostrobin, on Gibberella ear rot severity and mycotoxin production, both with and without WBC pressure, was also evaluated. The impact of each variable was assessed individually and in combination to determine the effect of each upon ear disease severity. RESULTS: There was a positive correlation between the presence of WBC larvae in field corn and Gibberella ear rot severity under inoculated conditions in the 2 years of the experiment. An application of pyraclostrobin did not impact Gibberella ear rot development when applied at corn growth stage R1 (silks first emerging). CONCLUSION: Feeding damage from WBC larvae significantly increases the development of F. graminearum in field corn. We conclude that an effective integrated management strategy for Gibberella ear rot should target the insect pest first, in an effort to limit disease severity and subsequent mycotoxin production by F. graminearum in kernels. © 2016 Society of Chemical Industry.


Assuntos
Gibberella/fisiologia , Mariposas/fisiologia , Doenças das Plantas/microbiologia , Zea mays/microbiologia , Animais , Comportamento Alimentar , Gibberella/crescimento & desenvolvimento , Indiana , Larva/fisiologia , Mariposas/crescimento & desenvolvimento , Zea mays/fisiologia
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