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1.
PLoS Pathog ; 16(9): e1008738, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32946515

RESUMO

Microsporidia, a divergent group of single-celled eukaryotic parasites, harness a specialized harpoon-like invasion apparatus called the polar tube (PT) to gain entry into host cells. The PT is tightly coiled within the transmissible extracellular spore, and is about 20 times the length of the spore. Once triggered, the PT is rapidly ejected and is thought to penetrate the host cell, acting as a conduit for the transfer of infectious cargo into the host. The organization of this specialized infection apparatus in the spore, how it is deployed, and how the nucleus and other large cargo are transported through the narrow PT are not well understood. Here we use serial block-face scanning electron microscopy to reveal the 3-dimensional architecture of the PT and its relative spatial orientation to other organelles within the spore. Using high-speed optical microscopy, we also capture and quantify the entire PT germination process of three human-infecting microsporidian species in vitro: Anncaliia algerae, Encephalitozoon hellem and E. intestinalis. Our results show that the emerging PT experiences very high accelerating forces to reach velocities exceeding 300 µm⋅s-1, and that firing kinetics differ markedly between species. Live-cell imaging reveals that the nucleus, which is at least 7 times larger than the diameter of the PT, undergoes extreme deformation to fit through the narrow tube, and moves at speeds comparable to PT extension. Our study sheds new light on the 3-dimensional organization, dynamics, and mechanism of PT extrusion, and shows how infectious cargo moves through the tube to initiate infection.


Assuntos
Microscopia/métodos , Microsporídios/patogenicidade , Organelas/imunologia , Organelas/ultraestrutura , Esporos Fúngicos/imunologia , Esporos Fúngicos/ultraestrutura , Proteínas Fúngicas/metabolismo , Microsporídios/imunologia , Microsporídios/ultraestrutura , Esporos Fúngicos/crescimento & desenvolvimento
2.
Nat Commun ; 11(1): 3897, 2020 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-32753587

RESUMO

Lipo-chitooligosaccharides (LCOs) are signaling molecules produced by rhizobial bacteria that trigger the nodulation process in legumes, and by some fungi that also establish symbiotic relationships with plants, notably the arbuscular and ecto mycorrhizal fungi. Here, we show that many other fungi also produce LCOs. We tested 59 species representing most fungal phyla, and found that 53 species produce LCOs that can be detected by functional assays and/or by mass spectroscopy. LCO treatment affects spore germination, branching of hyphae, pseudohyphal growth, and transcription in non-symbiotic fungi from the Ascomycete and Basidiomycete phyla. Our findings suggest that LCO production is common among fungi, and LCOs may function as signals regulating fungal growth and development.


Assuntos
Quitina/análogos & derivados , Quitina/metabolismo , Fungos/crescimento & desenvolvimento , Fungos/metabolismo , Transdução de Sinais/fisiologia , Ascomicetos/crescimento & desenvolvimento , Basidiomycota/crescimento & desenvolvimento , Ecologia , Ácidos Graxos/metabolismo , Micorrizas/fisiologia , Rhizobium/metabolismo , Esporos Fúngicos/crescimento & desenvolvimento , Simbiose/fisiologia
3.
PLoS One ; 15(8): e0233665, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32804955

RESUMO

Oligomycins are macrolide antibiotics, produced by Streptomyces spp. that show antagonistic effects against several microorganisms such as bacteria, fungi, nematodes and the oomycete Plasmopara viticola. Conidiogenesis, germination of conidia and formation of appressoria are determining factors pertaining to pathogenicity and successful diseases cycles of filamentous fungal phytopathogens. The goal of this research was to evaluate the in vitro suppressive effects of two oligomycins, oligomycin B and F along with a commercial fungicide Nativo® 75WG on hyphal growth, conidiogenesis, conidial germination, and appressorial formation of the wheat blast fungus, Magnaporthe oryzae Triticum (MoT) pathotype. We also determined the efficacy of these two oligomycins and the fungicide product in vivo in suppressing wheat blast with a detached leaf assay. Both oligomycins suppressed the growth of MoT mycelium in a dose dependent manner. Between the two natural products, oligomycin F provided higher inhibition of MoT hyphal growth compared to oligomycin B with a minimum inhibitory concentration of 0.005 and 0.05 µg/disk, respectively. The application of the compounds completely halted conidial formation of the MoT mycelium in agar medium. Further bioassays showed that these compounds significantly inhibited MoT conidia germination and induced lysis. The compounds also caused abnormal germ tube formation and suppressed appressorial formation of germinated spores. Interestingly, the application of these macrolides significantly inhibited wheat blast on detached leaves of wheat. This is the first report on the inhibition of mycelial growth, conidiogenesis, germination of conidia, deleterious morphological changes in germinated conidia, and suppression of blast disease of wheat by oligomycins from Streptomyces spp. Further study is needed to unravel the precise mode of action of these natural compounds and consider them as biopesticides for controlling wheat blast.


Assuntos
Magnaporthe/efeitos dos fármacos , Magnaporthe/patogenicidade , Oligomicinas/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Triticum/microbiologia , Agentes de Controle Biológico/farmacologia , Grão Comestível/microbiologia , Microbiologia de Alimentos , Fungicidas Industriais/farmacologia , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Magnaporthe/crescimento & desenvolvimento , Micélio/efeitos dos fármacos , Micélio/crescimento & desenvolvimento , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento
4.
Appl Environ Microbiol ; 86(17)2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32631867

RESUMO

Although dynamins and dynamin-related proteins (DRPs), a large GTPase superfamily, are involved in the budding of transport vesicles and division of organelles in eukaryotic cells, the function of these proteins in entomopathogenic fungi has not been reported to date. Here, DNM1, a DRP in Metarhizium robertsii, was characterized using gene disruption and complementation strategies. Mutant phenotype assays showed that the ΔDnm1 strain displayed increased defects in radial growth (∼24%) and conidial production (∼42%) compared to those of the wild type (WT), and reduced conidiation levels were accompanied by the repression of several key conidiation-related genes, including flbA, wetA, and flbD Additionally, mutant bioassays revealed that disruption of Dnm1 impaired the virulence (both topical inoculation and injection) of M. robertsii in the insect Galleria mellonella Further analysis demonstrated that deleting Dnm1 in fungi suppressed the transcriptional levels of several virulence genes in the insect hemocoel. Moreover, we found that DNM1 colocalized with peroxisomes and mitochondria. Importantly, disruption of Dnm1 abolished normal fungal endocytosis, resulting in significantly decreased numbers of, as well as morphological changes in, peroxisomes. These findings indicate that deletion of Dnm1 causes significant changes in the vegetative growth, sporulation, and virulence of M. robertsii due to changes in cell function and peroxisomes.IMPORTANCE Dnm1 was found to be involved in fungal development and virulence, mediated peroxisomal fission, and normal endocytosis. This finding provides new insights into the cellular processes and pathogenicity in entomopathogenic fungi.


Assuntos
Dinaminas/genética , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Metarhizium/fisiologia , Animais , Dinaminas/metabolismo , Endocitose/fisiologia , Proteínas Fúngicas/metabolismo , Metarhizium/genética , Metarhizium/crescimento & desenvolvimento , Metarhizium/patogenicidade , Mariposas/microbiologia , Peroxissomos/fisiologia , Controle Biológico de Vetores , Esporos Fúngicos/crescimento & desenvolvimento , Virulência
5.
Int J Food Microbiol ; 330: 108783, 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-32659523

RESUMO

The contamination of Aspergillus carbonarius causes decreases and great decay of agricultural products, and threatens the human and animal health by producing mycotoxins, especially ochratoxin A. Bacillus subtilis has been proved to efficiently inhibit the growth of A. carbonarius. Revealing the major active compound and the mechanisms for the antifungal of B. subtilis are essential to enhance its antifungal activity and control the quality of antifungal products made of it. In this study, we determined that iturin A is the major compound that inhibits Aspergillus carbonarius, a widespread fungal pathogen of grape and other fruits. Iturin A significantly inhibited growth and ochratoxin A production of A. carbonarius with minimal inhibitory concentrations (MICs) of 10 µg/mL and 0.312 µg/mL, respectively. Morphological observations revealed that iturin A caused swelling of the fungal cells and thinning of the cell wall and membrane at 1/2 MIC, whereas it inhibited fungal spore germination and caused mitochondrial swelling at higher concentrations. A differential transcriptomic analysis indicated that the mechanisms used by iturin A to inhibit A. carbonarius were to downregulate the expression of genes related to cell membrane, transport, osmotic pressure, oxidation-reduction processes, and energy metabolism. Among the down-regulated genes, those related to the transport capacity were most significantly influenced, including the increase of energy-related transport pathways and decrease of other pathways. Notably, the genes related to taurine and hypotaurine metabolism were also decreased, indicating iturin A potentially cause the occurrence of osmotic imbalance in A. carbonarius, which may be the intrinsic cause for the swelling of fungal cells and mitochondria. Overall, iturin A produced by B. subtilis played important roles to inhibit A. carbonarius via changing the fungal cell structure and causing perturbations to energy, transport and osmotic pressure metabolisms in fungi. The results indicated a new direction for researches on the mechanisms for lipopeptides and provided useful information to develop more efficient antifungal agents, which are important to agriculture and biomedicine.


Assuntos
Antifúngicos/farmacologia , Aspergillus/efeitos dos fármacos , Aspergillus/metabolismo , Bacillus subtilis/metabolismo , Peptídeos Cíclicos/farmacologia , Antifúngicos/metabolismo , Aspergillus/genética , Aspergillus/crescimento & desenvolvimento , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/genética , Metabolismo Energético/efeitos dos fármacos , Metabolismo Energético/genética , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Micotoxinas/metabolismo , Pressão Osmótica/efeitos dos fármacos , Peptídeos Cíclicos/metabolismo , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/metabolismo , Transcriptoma
6.
Int J Food Microbiol ; 333: 108781, 2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-32711130

RESUMO

Aspergillus fischeri ascospores are known as potential spoilage microorganisms of pasteurized fruit products due to their high incidence in fruits, the ability to survive pasteurization and to grow in acidic conditions. This study aimed to develop a quantitative microbial spoilage risk assessment (QMSRA) model approach to estimate the spoilage risk of packaged strawberry purees due to A. fischeri under various scenarios regarding product formulation, processing and storage conditions. The development of the risk assessment comprised three steps: (1) initial contamination level of raw material by ascospores (N0), (2) inactivation of ascospores during thermal processing (Np) and (3) determination of the number of ascospores which are able to survive thermal processing and develop visible mycelia (D = 2 mm) during storage (Nf). Data of visible growth (tv, days) comprised distributions previously obtained as function of water activity (aw) (0.860-0.985), oxygen (0-21%), temperature (8-30 °C) and pasteurization (95-105 °C/15 s). The simulations were performed in triplicate with 100,000 iterations using the software R. The outcome "spoilage risk" was defined as the probability of having at least one ascospore (Nf) capable of forming visible colonies in 100 g-pack strawberry puree within the typical use-by dates. Overall, high probabilities of spoilage were estimated for purees pasteurized at milder treatments at 85 °C/15-60 s (67%) and 90 °C/15-60 s (≥40%) stored at ambient temperature (22 °C). The spoilage risk was only effectively reduced (0.02%) by increasing pasteurization conditions to 95 °C for at least 45 s. Moreover, the microbial stability of such purees, i.e., spoilage risk <0.001% (=less than 1 spoilage pack out of 105 produced units) was predicted to occur for purees treated at 100 °C/15 s or stored at chilled conditions (≤8 °C) or at strict anaerobic conditions or produced as concentrates (aw ≤ 0.860). Based on the outcomes obtained, a set of specifications for Heat-Resistant Moulds (HRMs) in raw material and pasteurized purees aimed to be used as an ingredient was suggested. Furthermore, the results can be used to support risk management decisions in identifying and quantifying the impact of possible interventions during formulation, processing and storage conditions of fruit purees to effectively reduce this risk.


Assuntos
Aspergillus/metabolismo , Fragaria/microbiologia , Neosartorya/metabolismo , Medição de Risco/métodos , Esporos Fúngicos/crescimento & desenvolvimento , Aspergillus/crescimento & desenvolvimento , Contaminação de Alimentos , Microbiologia de Alimentos , Fragaria/metabolismo , Frutas/metabolismo , Frutas/microbiologia , Temperatura Alta , Neosartorya/crescimento & desenvolvimento , Pasteurização , Temperatura
7.
Int J Food Microbiol ; 328: 108687, 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32474227

RESUMO

Penicilium griseofulvum, the causal agent of apple blue mold, is able to produce in vitro and on apple a broad spectrum of secondary metabolites (SM), including patulin, roquefortine C and griseofulvin. Among them, griseofulvin is known for its antifungal and antiproliferative activity, and has received interest in many sectors, from medicine to agriculture. The biosynthesis of SM is finely regulated by filamentous fungi and can involve global regulators and pathway specific regulators, which are usually encoded by genes present in the same gene cluster as the backbone gene and tailoring enzymes. In the griseofulvin gene cluster, two putative transcription factors were previously identified, encoded by genes gsfR1 and gsfR2, and their role has been investigated in the present work. Analysis of P. griseofulvum knockout mutants lacking either gene suggest that gsfR2 forms part of a different pathway and gsfR1 exhibits many spectra of action, acting as regulator of griseofulvin and patulin biosynthesis and influencing conidia production and virulence on apple. The analysis of gsfR1 promoter revealed that the regulation of griseofulvin biosynthesis is also controlled by global regulators in response to many environmental stimuli, such as carbon and nitrogen. The influence of carbon and nitrogen on griseofulvin production was further investigated and verified, revealing a complex network of response and confirming the central role of gsfR1 in many processes in P. griseofulvum.


Assuntos
Griseofulvina/biossíntese , Patulina/biossíntese , Penicillium/metabolismo , Penicillium/patogenicidade , Esporos Fúngicos/crescimento & desenvolvimento , Carbono/metabolismo , Microbiologia de Alimentos , Griseofulvina/metabolismo , Malus/microbiologia , Família Multigênica , Nitrogênio/metabolismo , Patulina/metabolismo , Esporos Fúngicos/metabolismo , Fatores de Transcrição/genética , Virulência
8.
Int J Food Microbiol ; 326: 108649, 2020 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-32402917

RESUMO

In this study we assessed the efficacy of chitosan (CHI) (2%) emulsion added with Ruta graveolens L. essential oil (REO) at different concentrations (0.5%, 1.0% and 1.5%) to control C. gloesporioides grows both "in situ" and "in vitro" in papaya Maradol (Carica papaya L.). In vitro studies showed a decrease on fungal growth (mycelia diameter) with the increase of REO concentration, while 0.5% of REO induce a reduction of 56.42%, REO at 1.0% and 1.5% induced a reduction of 97%. Microscopic analysis showed irreversible deleterious morphological and ultrastructural alterations as well as changes in conidia morphology, and conidia germination inhibition up to 90%. Among the most abundant REO constituents, 2-Nonanol showed strong antifungal activity followed by 2-Undecanone, Benzyl acetate, 2-Nonanone, 2-Tridecanone and 2-Dodecanone. Studies "in situ" on papaya fruit during 12 days at 20 °C, showed a reduction of the C. gloesporioides lesion expansion by 50% using CHI-REO 0.5% emulsions and by 100% with treatments of CHI-REO 1.0 and 1.5%, in addition the emulsions were efficacious to reduce the fruit surface microbiota. On the other hand, physicochemical analysis of the papaya fruits demonstrated that CHI-REO emulsions treatment delayed papaya ripening without affecting the organoleptic characteristics. All these results demonstrated for the first time the application of coatings CHI-REO as a postharvest treatment for the control of anthracnose on papaya fruit.


Assuntos
Antifúngicos/farmacologia , Carica/microbiologia , Quitosana/farmacologia , Colletotrichum/efeitos dos fármacos , Óleos Voláteis/farmacologia , Ruta/metabolismo , Emulsões , Frutas/microbiologia , Micélio/crescimento & desenvolvimento , Esporos Fúngicos/crescimento & desenvolvimento
9.
Int J Food Microbiol ; 325: 108629, 2020 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-32325344

RESUMO

High pressure processing is a mild preservation process that inactivates pathogenic and spoilage micro-organisms in food products, but preserves the fresh characteristics of a product. Compared to untreated product, an enhanced shelf life is obtained during refrigerated storage. Knowledge on the use of high pressure pasteurisation aimed for ambient storage is limited. The aim of this research was to investigate if a combination of high pressure and moderate heat could be used to produce a shelf-stable high-acid fruit product. Ascospores of the heat resistant fungi Talaromyces macrosporus and Aspergillus fischeri were added to fresh strawberry puree that served as a model system. The effect of the processing steps and storage at ambient temperature for 2 weeks was studied on viability of the ascospores. A preheating step at 69 °C/2 min resulted in full or partial activation of A. fischeri and T. macrosporus spores, respectively. The pressure build-up by the process without any holding time resulted in additional activation of spores. A combination of moderate heat (maximum 85-90 °C) and high pressure (500-700 MPa) for holding times up to 13 min inactivated these highly resistant spores much faster than a heat treatment alone. At Tmax = 85 °C and 600 MPa the spores of T. macrosporus and A. fischeri were inactivated by 5.0 and 5.5 log10 after 13 and 7 min, respectively. At Tmax = 85 °C the heat treatment alone did not reduce the viability of these spores up to 60 min of treatment. At Tmax = 90 °C the holding time of the combined pressure-heat treatment could be reduced to obtain the same degree of inactivation of the heat resistant fungi. In addition, treated and untreated ascospores in strawberry puree were stored for 14 days at room temperature to evaluate delayed outgrowth of spores. Untreated ascospores of A. fischeri were activated by storage in the puree. However, at conditions combining high pressure ≥ 600 MPa with Tmax ≥ 85 °C for 13 min, heat resistant fungi were successfully inactivated. This research showed that a combination of moderate heat and pressure can drastically improve the effectiveness to inactivate heat-resistant ascospores in a high-acid fruit product compared to a heat treatment, potentially resulting in a better product quality.


Assuntos
Aspergillus/crescimento & desenvolvimento , Pasteurização/métodos , Esporos Bacterianos/crescimento & desenvolvimento , Esporos Fúngicos/crescimento & desenvolvimento , Talaromyces/crescimento & desenvolvimento , Microbiologia de Alimentos , Fragaria/microbiologia , Frutas/microbiologia , Temperatura Alta
10.
Int J Syst Evol Microbiol ; 70(5): 3162-3166, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32250241

RESUMO

An endophytic member of the genus Trichoderma was isolated from the root of a healthy 3-year-old Panax notoginseng in Yunnan province, PR China. The results of phylogenetic analyses based on a combined of ITS, tef1 and rpb2 indicated that this isolate was distinct from other species of the genus Trichoderma and closely related to Trichoderma songyi. It can be distinguished from T. songyi by its slower growth rates on PDA and colony morphology. The novel isolate formed conidia in thick white pustules scattered mostly at the margin. Its conidiophores tended to be regularly verticillium-like, little branched, sometimes substituted by phialides singly or in whorls. Conidia are smooth, mostly broadly subglobose to ellipsoidal. In combination with the genotypic and phenotypic characteristics, all data demonstrated that the fungus studied represented a unique and distinguishable novel species of the genus Trichoderma, for which the name Trichoderma panacis sp. nov. is proposed.


Assuntos
Panax notoginseng/microbiologia , Filogenia , Trichoderma/classificação , China , DNA Fúngico/genética , Endófitos/classificação , Endófitos/isolamento & purificação , Técnicas de Tipagem Micológica , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Esporos Fúngicos/crescimento & desenvolvimento , Trichoderma/isolamento & purificação
11.
Sci Rep ; 10(1): 3737, 2020 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-32111947

RESUMO

Encapsulated bioactive agents applied to the Lactuca sativa L. present an innovative approach to stimulate the production of plant secondary metabolites increasing its nutritive value. Calcium and copper ions were encapsulated in biopolymeric microparticles (microspheres and microcapsules) either as single agents or in combination with biocontrol agents, Trichoderma viride spores, a fungal plant growth mediator. Both, calcium and copper ions are directly involved in the synthesis of plant secondary metabolites and alongside, Trichoderma viride can provide indirect stimulation and higher uptake of nutrients. All treatments with microparticles had a positive effect on the enhancement of plant secondary metabolites content in Lactuca sativa L. The highest increase of chlorophylls, antioxidant activity and phenolic was obtained by calcium-based microparticles in both, conventionally and hydroponically grown lettuces. Non-encapsulated fungus Trichoderma viride enhanced the synthesis of plant secondary metabolites only in hydroponics cultivation signifying the importance of its encapsulation. Encapsulation proved to be simple, sustainable and environmentally favorable for the production of lettuce with increased nutritional quality, which is lettuce fortified with important bioactive compounds.


Assuntos
Cálcio/farmacologia , Cobre/farmacologia , Alface , Doenças das Plantas/microbiologia , Folhas de Planta , Esporos Fúngicos/crescimento & desenvolvimento , Trichoderma/crescimento & desenvolvimento , Cápsulas , Alface/metabolismo , Alface/microbiologia , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia
12.
Sci Rep ; 10(1): 3673, 2020 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-32111863

RESUMO

Fungal diseases seriously affect agricultural production and the food industry. Crop protection is usually achieved by synthetic fungicides, therefore more sustainable and innovative technologies are increasingly required. The atmospheric pressure low-temperature plasma is a novel suitable measure. We report on the effect of plasma treatment on phytopathogenic fungi causing quantitative and qualitative losses of products both in the field and postharvest. We focus our attention on the in vitro direct inhibitory effect of non-contact Surface Dielectric Barrier Discharge on conidia germination of Botrytis cinerea, Monilinia fructicola, Aspergillus carbonarius and Alternaria alternata. A few minutes of treatment was required to completely inactivate the fungi on an artificial medium. Morphological analysis of spores by Scanning Electron Microscopy suggests that the main mechanism is plasma etching due to Reactive Oxygen Species or UV radiation. Spectroscopic analysis of plasma generated in humid air gives the hint that the rotational temperature of gas should not play a relevant role being very close to room temperature. In vivo experiments on artificially inoculated cherry fruits demonstrated that inactivation of fungal spores by the direct inhibitory effect of plasma extend their shelf life. Pre-treatment of fruits before inoculation improve the resistance to infections maybe by activating defense responses in plant tissues.


Assuntos
Fungos Mitospóricos/crescimento & desenvolvimento , Doenças das Plantas/microbiologia , Gases em Plasma , Esporos Fúngicos/crescimento & desenvolvimento , Gases em Plasma/química , Gases em Plasma/farmacologia
13.
Arch Microbiol ; 202(6): 1459-1467, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32189017

RESUMO

Sunflower is one of the most economically important oil crops. Recently, sunflower anthracnose caused by Colletotrichum destructivum was reported and suggested to be a potential threat to the quality of oil and edible seeds derived from sunflower in the field and even on the ornamentals in the residential gardens. Colletotrichum destructivum, as the causal agent of sunflower anthracnose, has been rarely studied. In this study, the vegetative growth and sporulation of this fungal species were investigated by assessing the requirements of nutrition and other environmental conditions, such as temperature, ambient pH, and lightness regime. Additionally, the sensitivity of C. destructivum to several fungicides was assessed. The results will provide a baseline for better understanding of the biology and etiology of C. destructivum. This study will be the first reference for a sustainable management strategy according to the occurrence and prevalence of the sunflower anthracnose.


Assuntos
Colletotrichum/crescimento & desenvolvimento , Fungicidas Industriais/farmacologia , Helianthus/microbiologia , Doenças das Plantas/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Colletotrichum/classificação , Colletotrichum/efeitos dos fármacos , Colletotrichum/isolamento & purificação
14.
World J Microbiol Biotechnol ; 36(4): 57, 2020 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-32219579

RESUMO

Although microbial transformation has been expected as a substitution technology for organic synthesis, microbial toxicity and water-insolubility of synthetic substrates prevent the practical application of the technology. For these problems, the authors have developed two types of interfacial bioprocesses, solid-liquid and liquid-liquid interface bioreactors and applied the systems to many microbial transformations. In the bioreactors, addition of substrates and accumulation of products were remarkably enhanced based on the toxicity alleviation effect on the interfaces and solubilization of substrates and/or products in an organic phase of the bioreactors. Recently, a novel tacky liquid-liquid interface bioreactor has been developed and applied to actinomycetes and yeasts. Furthermore, a novel bioconversion system with fungal spores in an organic phase has been constructed based on the combination of two facts as follows: (i) the fungal spores are never resting cells and are active ones like the vegetable cells, (ii) the fungal spores have the excellent solvent-tolerance. In this review, the tacky liquid-liquid interface bioreactor (L-L IBRtac) and the non-aqueous sporular bioconversion system with immobilized fungal spores (NASB) are mainly given outlines.


Assuntos
Reatores Biológicos/microbiologia , Poluentes Químicos da Água/química , Bactérias/crescimento & desenvolvimento , Biodegradação Ambiental , Fungos/crescimento & desenvolvimento , Solventes , Esporos Fúngicos/crescimento & desenvolvimento
15.
PLoS Pathog ; 16(2): e1008355, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32092131

RESUMO

Genetic studies have shown essential functions of N-glycosylation during infection of the plant pathogenic fungi, however, systematic roles of N-glycosylation in fungi is still largely unknown. Biological analysis demonstrated N-glycosylated proteins were widely present at different development stages of Magnaporthe oryzae and especially increased in the appressorium and invasive hyphae. A large-scale quantitative proteomics analysis was then performed to explore the roles of N-glycosylation in M. oryzae. A total of 559 N-glycosites from 355 proteins were identified and quantified at different developmental stages. Functional classification to the N-glycosylated proteins revealed N-glycosylation can coordinate different cellular processes for mycelial growth, conidium formation, and appressorium formation. N-glycosylation can also modify key components in N-glycosylation, O-glycosylation and GPI anchor pathways, indicating intimate crosstalk between these pathways. Interestingly, we found nearly all key components of the endoplasmic reticulum quality control (ERQC) system were highly N-glycosylated in conidium and appressorium. Phenotypic analyses to the gene deletion mutants revealed four ERQC components, Gls1, Gls2, GTB1 and Cnx1, are important for mycelial growth, conidiation, and invasive hyphal growth in host cells. Subsequently, we identified the Gls1 N-glycosite N497 was important for invasive hyphal growth and partially required for conidiation, but didn't affect colony growth. Mutation of N497 resulted in reduction of Gls1 in protein level, and localization from ER into the vacuole, suggesting N497 is important for protein stability of Gls1. Our study showed a snapshot of the N-glycosylation landscape in plant pathogenic fungi, indicating functions of this modification in cellular processes, developments and pathogenesis.


Assuntos
Retículo Endoplasmático/metabolismo , Magnaporthe/genética , Magnaporthe/metabolismo , Estudos de Avaliação como Assunto , Proteínas Fúngicas/metabolismo , Deleção de Genes , Regulação Fúngica da Expressão Gênica/genética , Genes Fúngicos/genética , Glicosilação , Hifas/genética , Mutação , Oryza/microbiologia , Doenças das Plantas/microbiologia , Proteômica/métodos , Deleção de Sequência , Esporos Fúngicos/crescimento & desenvolvimento , Virulência/genética
16.
J Appl Microbiol ; 129(2): 278-286, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32097516

RESUMO

AIMS: The aim of the study was to investigate the antifungal effects of a quorum sensing-molecule, 2-phenylethanol, against the food spoilage moulds Penicillium expansum and Penicillium nordicum. METHODS AND RESULTS: Conidial germination of the tested Penicillium spp. (three strains in total) were inhibited by treatments with 2-phenylethanol in a concentration-dependent manner. Germinated conidia was significantly reduced from 4·4-16·7% at 7·5 mmol l-1 and completely inhibited at 15 mmol l-1 2-phenylethanol. Integrity of conidial cell membranes was unaffected by 2-phenylethanol resulting in reversible inhibition pattern of germination. In contrast, membrane permeability of actively growing hyphae was severely compromised, showing 63·5 - 75·7% membrane damage upon treatment with 15 mmol l-1 2-phenylethanol. The overall inhibitory effect of 2-phenylethanol on colony development and growth of P. expansum and P. nordicum was additionally confirmed. CONCLUSIONS: 2-phenylethanol inhibits conidial germination and growth of P. expansum and P. nordicum in a nonlethal, reversible and concentration-dependent manner. SIGNIFICANCE AND IMPACT OF THE STUDY: The study indicates that 2-phenylethanol can find potential application as an antifungal agent for biological control of moulds in the food industry.


Assuntos
Antifúngicos/farmacologia , Penicillium/efeitos dos fármacos , Álcool Feniletílico/farmacologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Hifas/efeitos dos fármacos , Penicillium/crescimento & desenvolvimento , Percepção de Quorum , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento
17.
J Appl Microbiol ; 129(2): 400-410, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32043673

RESUMO

AIMS: To isolate and characterize a native strain of Trichothecium roseum infecting the immatures of Pauropsylla buxtoni on fig leaves, to study the morphological features of the isolated strain, then to test the entomopathogenic effect of the isolated strain against the immatures of P. buxtoni on fig leaves. METHODS AND RESULTS: The isolated strain of T. roseum produced pink mycelial growth on culture medium with septate mycelium and conidiophores. It also produced two-celled conidia with elliptical to pyriform shape born at the tip of conidiophores. Molecular characterization of the isolated strain confirmed the identity of the strain as T. roseum. In bioassays, application of conidial suspension of the isolated strain against the 4th instar of P. buxtoni immatures infesting fig leaves showed an obvious entomopathogenic effect of the applied fungus strain against the targeted insect. This effect was exhibited by the death of treated P. buxtoni immatures with the fungus. The dead insects were characterized by the presence of pinkish mycelial growth on the outer surface which is characteristic to the fungus, in addition to the positive isolation of the fungus from internal tissues of treated insects after a proper external disinfection. Moreover, significant differences (at P < 0·018) were obtained between the means of mortality % of P. buxtoni immatures treated with different concentrations of conidial suspension of the fungus. CONCLUSIONS: The overall results confirm the entomopathogenic effect of T. roseum against P. buxtoni immatures infesting fig leaves. Significant mortalities of P. buxtoni immatures were obtained when the different concentrations of the fungus conidial suspension were bio-assessed against the insect. SIGNIFICANCE AND IMPACT OF THE STUDY: The tested strain of T. roseum can be applied as biocontrol agent of P. buxtoni on fig leaves within an integrated control programme to reduce the impact of pest on fig trees.


Assuntos
Ficus/parasitologia , Hemípteros/microbiologia , Hypocreales/patogenicidade , Controle Biológico de Vetores/métodos , Doenças das Plantas/prevenção & controle , Animais , Hemípteros/crescimento & desenvolvimento , Hypocreales/classificação , Hypocreales/citologia , Hypocreales/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Larva/microbiologia , Doenças das Plantas/parasitologia , Folhas de Planta/parasitologia , Esporos Fúngicos/classificação , Esporos Fúngicos/citologia , Esporos Fúngicos/crescimento & desenvolvimento
18.
J Appl Microbiol ; 129(2): 411-421, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32086853

RESUMO

AIMS: Mitochondrial fission is an essential dynamics that maintains mitochondrial morphology and function. This study seeks to determine the roles of mitochondrial fission in the filamentous entomopathogenic fungus Beauveria bassiana. MATERIAL AND METHODS: Three fission-related genes (BbFis1, BbMdv1 and BbDnm1) were functionally characterized via protein intracellular localization and construction of gene disruption mutants. RESULTS: Mitochondrial localization was only observed for BbFis1 which interacts with BbMdv1, but BbMdv1 did not have interaction with BbDnm1. Single disruption mutant of three genes generated the elongated and enlarged mitochondria which could not be eliminated via the mitophagy. Three mutant strains displayed the reduced ATP synthesis and vegetative growth compared with the wild type. Three genes were involved in the early stage of conidiation and unnecessary for the late stage. However, all three genes significantly contribute to blastospore development under submerged condition, and the loss of BbMdv1 had the greatest effects compared with the losses of BbFis1 or BbDnm1. Finally, disruption of three genes significantly attenuated fungal virulence, but their mutations had different influences. CONCLUSIONS: In addition to their consistent roles in mitochondrial division and mitophagy, three fission-related genes perform divergent roles in the development and virulence of the entomopathogenic fungus B. bassiana. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that mitochondrial fission is associated with lifecycle of B. bassiana. These findings provide information for the manipulation of fungal physiology and facilitate the application of entomopathogenic fungi.


Assuntos
Beauveria/crescimento & desenvolvimento , Beauveria/patogenicidade , Insetos/microbiologia , Dinâmica Mitocondrial/fisiologia , Mitofagia , Animais , Beauveria/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Dinâmica Mitocondrial/genética , Mitofagia/genética , Esporos Fúngicos/crescimento & desenvolvimento , Virulência/genética
19.
Appl Environ Microbiol ; 86(7)2020 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-32005734

RESUMO

Aspergillus fumigatus is a major cause of human disease. The survival of this fungus is dependent on the cell wall organization and function of its components. The cell wall integrity pathway (CWIP) is the primary signaling cascade that controls de novo synthesis of the cell wall in fungi. Abundant conidiation is a hallmark in A. fumigatus, and uptake of conidia by a susceptible host is usually the initial event in infection. The formation of conidia is mediated by the development of fungus-specific specialized structures, conidiophores, which are accompanied by cell wall remodeling. The molecular regulation of these changes in cell wall composition required for the rise of conidiophore from the solid surface and to disperse the conidia into the air is currently unknown. Here, we investigated the role of CWIP in conidiation. We show that CWIP pkcA G579R, ΔmpkA, and ΔrlmA mutants displayed reduced conidiation during synchronized asexual differentiation. The transcription factor RlmA directly regulated the expression of regulators of conidiation, including flbB, flbC, brlA, abaA, and rasB, as well as genes involved in cell wall synthesis and remodeling, and this affected the chitin content in aerial hyphae. Phosphorylation of RlmA and MpkA was increased during asexual differentiation. We also observed that MpkA physically associated with the proteins FlbB, FlbC, BrlA, and RasB during this process, suggesting another level of cross talk between the CWIP and asexual development pathways. In summary, our results support the conclusion that one function of the CWIP is the regulation of asexual development in filamentous fungi.IMPORTANCE A remarkable feature of the human pathogen Aspergillus fumigatus is its ability to produce impressive amounts of infectious propagules known as conidia. These particles reach immunocompromised patients and may initiate a life-threatening mycosis. The conidiation process in Aspergillus is governed by a sequence of proteins that coordinate the development of conidiophores. This process requires the remodeling of the cell wall so that the conidiophores can rise and withstand the chains of conidia. The events regulating cell wall remodeling during conidiation are currently unknown. Here, we show that the cell wall integrity pathway (CWIP) components RlmA and MpkA directly contribute to the activation of the conidiation cascade by enabling transcription or phosphorylation of critical proteins involved in asexual development. This study points to an essential role for the CWIP during conidiation and provides further insights into the complex regulation of asexual development in filamentous fungi.


Assuntos
Aspergillus fumigatus/fisiologia , Parede Celular/metabolismo , Proteínas Fúngicas/metabolismo , Reprodução Assexuada , Transdução de Sinais , Esporos Fúngicos/crescimento & desenvolvimento , Aspergilose/microbiologia , Aspergillus fumigatus/crescimento & desenvolvimento , Proteínas Fúngicas/genética , Humanos
20.
Sci Rep ; 10(1): 102, 2020 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-31919462

RESUMO

Basidioradulum was morphologically considered to be a synonym of Xylodon. Here, its independence within Hymenochaetales is confirmed from a phylogenetic perspective. Basidioradulum radula, the generic type, is widely distributed in Northern Hemisphere. Two Southern Hemisphere species close to B. radula are newly described as B. mayi and B. tasmanicum, respectively, from Victoria and Tasmania, Australia. Basidioradulum mayi differs from B. radula by lack of cystidia. Moreover, the hymenial surface of B. radula is normally much more strongly hydnoid than that of B. mayi. Basidioradulum tasmanicum is distinct from B. radula and B. mayi by having capitate cystidia, ellipsoid to subglobose basidiospores, and crystal-covered hyphae. Although morphologically distinct, the two new species isolated by Bass Strait have an almost identical ITS region, and could not be differentiated by nLSU- and ITS-based phylogenetic analyses. This case reminds us that basing phylogeny simply on the ITS as a barcode region may underestimate fungal species diversity.


Assuntos
Basidiomycota/classificação , Basidiomycota/genética , DNA Fúngico/análise , DNA Espaçador Ribossômico/genética , RNA Ribossômico 28S/análise , Esporos Fúngicos/genética , Austrália , Basidiomycota/crescimento & desenvolvimento , DNA Fúngico/genética , Filogenia , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Esporos Fúngicos/crescimento & desenvolvimento
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