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1.
J Agric Food Chem ; 68(1): 185-192, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31815467

RESUMO

Phytopathogenic fungi have been considered as being an enormous threat in the agricultural system. In our search of new antifungal natural products, nine new halogenated cyclopentenones, bicolorins A-I (1-3, and 5-10), along with three known cyclopentenones (4, 11, and 12) were isolated from the endophytic fungus Saccharicola bicolor of Bergenia purpurascens by the one strain-many compounds strategy. Their structures and absolute configurations were elucidated based on extensive spectroscopic analysis, X-ray crystallographic analysis, and time-dependent density functional theory-equivalent circulating density calculations. Compounds 1-12 showed antifungal activities against five phytopathogenic fungi (Uromyces viciae-fabae, Pythium dissimile, Gibberella zeae, Aspergillus niger, and Sclerotinia sclerotiorum). Especially, bicolorins B and D (2 and 5) exhibited strong antifungal activities against P. dissimile with the MIC values of 6.2 and 8.5 µg/mL, respectively, compared with the positive control cycloheximide (MIC of 8.6 µg/mL). Additionally, bicolorin D was proven to be potently antifungal against S. sclerotiorum in vitro and in vivo. This work provides an effective strategy for searching antifungal candidate agents.


Assuntos
Ascomicetos/química , Ciclopentanos/química , Ciclopentanos/farmacologia , Endófitos/química , Fungicidas Industriais/química , Fungicidas Industriais/farmacologia , Saxifragaceae/microbiologia , Ascomicetos/isolamento & purificação , Ascomicetos/metabolismo , Ciclopentanos/metabolismo , Endófitos/isolamento & purificação , Endófitos/metabolismo , Fungos/efeitos dos fármacos , Fungos/crescimento & desenvolvimento , Fungicidas Industriais/metabolismo , Halogenação , Estrutura Molecular , Doenças das Plantas/microbiologia
2.
Chemosphere ; 238: 124700, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31524602

RESUMO

An eight-year field trial was conducted to investigate the effects of four different N fertilization treatments of urea (CO(NH2)2, the control), ammonium sulfate ((NH4)2SO4), ammonium chloride (NH4Cl), and ammonium hydrogen phosphate [(NH4)2HPO4]) on cadmium (Cd) phytotoxicity in rice and soil microbial communities in a Cd-contaminated paddy of southern China. The results demonstrate that the different N treatments exerted different effects: the application of (NH4)2HPO4 and (NH4)2SO4 significantly increased rice grain yield and decreased soil-extractable Cd content when compared with those of the control, while NH4Cl had a converse effect. Expression of genes related to Cd uptake (IRT and NRAPM genes) and transport (HMA genes) by roots may be responsible for Cd phytotoxicity in rice grown in the different N fertilization treatments. Our results further demonstrate that N fertilization had stronger effects on soil bacterial communities than fungal communities. The bacterial and fungal keystone species were identified by phylogenetic molecular ecological network (pMEN) analysis and mainly fell into the categories of Gammaproteobacteria, Acidobacteria and Actinobacteria for the bacterial species and Ascomycota for the fungal species; all of these keystone species were highly enriched in the (NH4)2HPO4 treatment. Soil pH and soil available-Cd content emerged as the major determinants of microbial network connectors. These results could provide effective fertilizing strategies for alleviating Cd phytotoxicity in rice and enhance the understanding of its underlying microbial mechanisms.


Assuntos
Ascomicetos/isolamento & purificação , Bactérias/isolamento & purificação , Cádmio/análise , Cádmio/toxicidade , Oryza/química , Poluentes do Solo/toxicidade , Acidobacteria/isolamento & purificação , Actinobacteria/isolamento & purificação , Cloreto de Amônio/química , Sulfato de Amônio/química , Ascomicetos/metabolismo , Bactérias/metabolismo , China , Grão Comestível/química , Poluição Ambiental/análise , Fertilizantes/análise , Gammaproteobacteria/isolamento & purificação , Microbiota , Oryza/microbiologia , Fosfatos/química , Filogenia , Raízes de Plantas/metabolismo , Raízes de Plantas/microbiologia , Solo/química , Microbiologia do Solo , Poluentes do Solo/análise , Ureia/química
3.
An Bras Dermatol ; 94(5): 574-577, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31780436

RESUMO

The authors report a case of 40-year-old male patient with a five-year history of chromoblastomycosis on his right leg. Diagnosis was performed by direct 40% KOH exam of skin scales, culture with micro- and macromorphologic analysis, and genotypic characterization (sequencing of a fragment of the ITS region and phylogenetic analysis) of the isolated fungus. Rhinocladiella aquaspersa was identified as the etiological agent. Initially, the treatment was with oral itraconazole 200mg/day for one year. However, the presence of "sclerotic cells" with filaments ("Borelli spiders") resulted in a change of medical treatment: a higher dose of itraconazole (400mg/day) and surgery, achieving clinical and mycological cure in one year. This is the first report of chromoblastomycosis caused by R. aquaspersa in Guatemala.


Assuntos
Ascomicetos/isolamento & purificação , Cromoblastomicose/microbiologia , Adulto , Antifúngicos/uso terapêutico , Ascomicetos/ultraestrutura , Cromoblastomicose/tratamento farmacológico , Cromoblastomicose/patologia , Guatemala , Humanos , Itraconazol/uso terapêutico , Masculino , Resultado do Tratamento
4.
BMC Vet Res ; 15(1): 354, 2019 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-31639008

RESUMO

BACKGROUND: Mycoleptodiscus indicus is a dematiaceous hyphomycete fungus found on plant leaves. It has been rarely reported as a cause of human or animal disease, possibly because it is difficult to culture and identify from clinical specimens. Infections are presumably acquired by traumatic implantation. CASE PRESENTATION: An 8-year-old non-immunosuppressed cat from Georgia, USA, presented with a left front leg swelling without lameness. Cytology from a fine needle aspirate revealed pyogranulomatous inflammation with both cytoplasmic and extracellular fungal elements. There were septate hyphae with irregularly sized segments, non-staining uneven walls, and rounded yeast-like forms from which longer hyphae arose in a hub-and-spoke pattern. A mold was isolated on agar from a fine needle aspirate collected 1 week later and identified as M. indicus by morphology, DNA sequencing and phylogenetic analysis. The cat recovered completely and uneventfully with antifungal treatment. CONCLUSIONS: We report a previously undescribed presentation of M. indicus causing a subcutaneous infection in a cat with successful antifungal treatment. In this study we highlight the potential of M. indicus to infect immunocompetent animals, and the veterinary medical community should be aware of its unusual but characteristic clinical, microbiological and cytologic presentation.


Assuntos
Ascomicetos , Doenças do Gato/microbiologia , Micoses/veterinária , Infecções dos Tecidos Moles/veterinária , Animais , Antifúngicos/uso terapêutico , Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Doenças do Gato/imunologia , Gatos , Fluconazol/uso terapêutico , Membro Anterior , Imunocompetência , Masculino , Micoses/imunologia , Filogenia , Infecções dos Tecidos Moles/imunologia , Infecções dos Tecidos Moles/microbiologia , Tela Subcutânea , Resultado do Tratamento
5.
Plant Dis ; 103(12): 3129-3141, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31642736

RESUMO

Canker and wood rot pathogens cause dieback and, in severe cases, the death of young apple trees. Recently, a higher occurrence of cankers was observed on 1-year-old apple trees in the Western Cape Province of South Africa. This study aimed to assess the phytosanitary status of nursery trees and propagation material as possible inoculum sources for canker pathogens. Thirteen 1-year-old apple orchards showing canker or dieback symptoms were sampled. Certified nursery apple trees were collected from four nurseries as well as scion and rootstock mother plant material. Isolations were made from the discoloration observed in the vascular tissue of the plant parts and from asymptomatic material. Possible canker and wood rot species were identified with PCR and sequence comparisons of the relevant gene regions and phylogenetic analyses. Similar canker and wood rot species were isolated from 1-year-old diseased apple trees, nursery apple trees, and the propagation material. Forty-five fungal species associated with canker or wood rot symptoms were identified. The top five most abundant fungal species found causing disease on commercial 1-year-old trees were also found in high numbers causing latent infection in certified apple nursery trees. These species were Didymosphaeria rubi-ulmifolii sensu lato, Diplodia seriata, Schizophyllum commune, Didymella pomorum, and Coniochaeta fasciculata, with D. rubi-ulmifolii sensu lato being the dominant species in both sampling materials. In all, 65% of certified nursery apple trees, 5% of scion shoots used for budding, and 21% of rooted rootstock cuttings from layer blocks had latent infections of canker and wood rot pathogens. Pathogenicity trials were conducted with isolates of 39 species, inoculated onto 2-year-old branches of 14-year-old Golden Delicious trees. All species caused lesions that were significantly longer than the control. This study confirmed the presence of canker and wood rot pathogens in apple propagation material as well as certified nursery apple trees, which will aid the improvement of management practices in nurseries.


Assuntos
Ascomicetos , Malus , Doenças das Plantas , Ascomicetos/classificação , Ascomicetos/genética , Ascomicetos/isolamento & purificação , Malus/microbiologia , Filogenia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , África do Sul , Madeira/microbiologia
6.
BMC Infect Dis ; 19(1): 843, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31615438

RESUMO

BACKGROUND: In recent years, some rare fungi have been increasingly recognized as new human pathogens. Here we reported the first fatal case of human severe pneumonia complicated by multiple organ dysfunction caused by Acrophialophora levis infection. However, its pathogenic mechanism and risk factors are unknown. Acrophialophora genus has only reported in six cases of human infection worldwide, but it has not been reported previously in China. CASE PRESENTATION: A 71-year-old male patient with severe pneumonia complicated with multiple organ dysfunction caused by A. levis infection. The fungal identification was based on micromorphology and sequence analysis of the internal transcriptional spacer (ITS) of ribosomal RNA genes recovered from lower respiratory tract secretions. The microbial characteristics, sensitivity to antifungal drugs of this isolated A. levis were studied. Anti-infective regimen, liposomal amphotericin B combined with tegacycline, was used to prevent infection. The next day, the fever decreased, body temperature fluctuated between 36.5 and 37.8 degree, cough and sputum decreased, and sputum volume decreased, with oxygen uptake for 5 L/min, blood oxygen saturation over 95%. After 17 days of treatment, CT reexamination showed that the lesions in the right lung and left upper lung were absorbed and pleural effusion was reduced. The next 8 days, the patient asked to return to the local hospital for treatment. The local hospital stopped using liposomal amphotericin B because of the absence of liposomal amphotericin B, and died of respiratory failure 2 days later. CONCLUSIONS: This study is the first to report the occurrence, risk factors, molecular determinants, microbial characteristics and susceptibility to antifungal agents of A. levis infection in China. In addition, six published cases of human infection with Acrophialophora were reviewed.


Assuntos
Ascomicetos/isolamento & purificação , Pneumonia/diagnóstico , Idoso , Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Humanos , Masculino , Derrame Pleural/etiologia , Pneumonia/complicações , Pneumonia/tratamento farmacológico , Pneumonia/patologia , Índice de Gravidade de Doença , Tomografia Computadorizada por Raios X
7.
Chem Biodivers ; 16(12): e1900519, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31596057

RESUMO

A new pimarane diterpenoid, named botryopimarene A (1), was discovered from the fungus Botryotinia fuckeliana MCCC 3 A00494 isolated from the deep-sea water, together with ten known compounds. The planar structure of 1 was established based on the extensive spectroscopic analyses. The absolute configurations of tricyclic system in 1 were resolved by the theoretical ECD calculation, while the 15,16-diol moiety in the side chain was resolved by the Mo2 (OAc)4 -induced ECD spectrum. Compound 1, featuring a Δ9(11) double bond, was rarely discovered in pimarane family. Compounds 1-11 were tested for their cytotoxic activities using six human cancer cell lines by the MTT method. However, none of the compounds exhibited detectable cytotoxicities (IC50 >20 µm).


Assuntos
/química , Ascomicetos/química , Diterpenos/química , Água do Mar/microbiologia , /isolamento & purificação , Ascomicetos/isolamento & purificação , Ascomicetos/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Dicroísmo Circular , Diterpenos/isolamento & purificação , Diterpenos/farmacologia , Humanos , Conformação Molecular
8.
Zhongguo Zhong Yao Za Zhi ; 44(15): 3213-3220, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31602874

RESUMO

A total of 27 endophytic fungal strains were isolated from Huperzia serrata,which were richly distributed in the stems and leaves while less distributed in roots. The 27 strains were identified by Internal Transcribed Spacer( ITS) r DNA molecular method and one of the strains belongs to Basidiomycota phylum,and other 26 stains belong to 26 species,9 general,6 families,5 orders,3 classes of Ascomycota Phylum. The dominant strains were Colletotrichum genus,belonging to Glomerellaceae family,Glomerellales order,Sordariomycetes class,Ascomycota Phylum,with the percentage of 48. 15%. The inhibitory activities of the crude extracts of 27 endophytic fungal strains against acetylcholinesterase( ACh E) and nitric oxide( NO) production were evaluated by Ellman's method and Griess method,respectively. Crude extracts of four fungi exhibited inhibitory activities against ACh E with an IC50 value of 42. 5-62. 4 mg·L~(-1),and some fungi's crude extracts were found to inhibit nitric oxide( NO) production in lipopolysaccharide( LPS)-activated RAW264. 7 macrophage cells with an IC50 value of 2. 2-51. 3 mg·L~(-1),which indicated that these fungi had potential anti-inflammatory activities.The chemical composition of the Et OAc extract of endophytic fungus HS21 was also analyzed by LCMS-IT-TOF. Seventeen compounds including six polyketides,four diphenyl ether derivatives and seven meroterpenoids were putatively identified.


Assuntos
Ascomicetos/química , Ascomicetos/classificação , Huperzia/microbiologia , Acetilcolinesterase , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Ascomicetos/isolamento & purificação , Inibidores da Colinesterase/isolamento & purificação , Inibidores da Colinesterase/metabolismo , Endófitos/classificação , Endófitos/isolamento & purificação , Camundongos , Células RAW 264.7
9.
Biofouling ; 35(8): 856-869, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31603000

RESUMO

Microbial contamination during fuel storage can cause fuel system fouling and corrosion. Characterizing microbial contamination is critical for preventing and solving these problems. In this study, culture-based combing with the culture-independent methods, were used to profile the microbial contamination in aviation fuel. High-throughput sequencing (HTS) modified by propidium monoazide (PMA) revealed a higher diversity of contaminating microorganisms in samples than the culture method. Proteobacteria (47%), Actinobacteria (21%) and Ascomycota (>99%, fungi) were the most abundant phyla, and the neglected archaea was also detected. Additionally, qPCR-based methods revealed all samples contained a heavy level of microbial contamination, which was more accurate than its culturable counterparts, and fungal contamination was still a problem in aviation fuel. The application of a PCR-based method gives deeper insight into microbial contamination in aviation fuel than the conventional culture method, thus using it for regular detection and accurate description of fuel contamination is strongly recommended in the case of explosive microbial growth.


Assuntos
Actinobacteria/isolamento & purificação , Aeroportos , Ascomicetos/isolamento & purificação , Incrustação Biológica/prevenção & controle , Gasolina/microbiologia , Proteobactérias/isolamento & purificação , Gasolina/normas , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota/genética , RNA Ribossômico 16S , Reação em Cadeia da Polimerase em Tempo Real
10.
Lett Appl Microbiol ; 69(5): 358-365, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31508838

RESUMO

Leaf blight and fruit rot disease caused by Phomopsis vexans is a devastating disease of brinjal. The detection of P. vexans in plant parts and seeds of brinjal can be complicated, mainly when the inoculum is present at low levels and/or overgrown by fast-growing saprophytic fungi or other seed-borne fungi. A PCR-based diagnostic method was developed with specific primers designed based on sequence data of a region consisting of the 5·8S RNA gene and internal transcribed spacers, ITS 1 and ITS 2 of nuclear ribosomal RNA gene (rDNA) repeats of P. vexans. The efficiency and specificity of primer pairs PvexF/PvexR designed were established by PCR analysis of DNA from P. vexans strains isolated from India and fungal isolates of other genera. A single amplification product of 323-bp was detected from DNA of P. vexans isolates. No cross-reaction was observed with any of the other isolates tested. The specific primers designed and employed in PCR detected P. vexans up to 10 pg from DNA isolated from pure culture. This is the first report on the development of species-specific PCR assay for identification and detection of P. vexans. Thus, PCR-based assay developed is very specific, rapid, confirmatory and sensitive tool for the detection of pathogen P. vexans at early stages. SIGNIFICANCE AND IMPACT OF THE STUDY: Phomopsis vexans is an important seed-borne pathogenic fungus responsible for leaf blight and fruit rot in brinjal. Current detection methods, based on culture and morphological identification is time consuming, laborious and are not always reliable. A PCR-based diagnostic method was developed with species-specific primers designed based on sequence data of a region consisting of the 5·8S RNA gene and internal transcribed spacers, ITS 1 and ITS 2 of nuclear ribosomal RNA gene (rDNA) repeats of P. vexans.


Assuntos
Ascomicetos/isolamento & purificação , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase/métodos , Solanum melongena/microbiologia , Ascomicetos/classificação , Ascomicetos/genética , Primers do DNA/genética , DNA Fúngico/genética , DNA Ribossômico/genética , Frutas/microbiologia , Índia , Folhas de Planta/microbiologia , Sementes/microbiologia
11.
BMC Evol Biol ; 19(1): 183, 2019 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533625

RESUMO

BACKGROUND: Blumeria graminis f. sp. tritici (Bgt), the causal agent of wheat powdery mildew severely affects yield security wheat production in China. Understanding the virulence structure and genetic variations of this pathogen is important for breeding wheat lines resistant to wheat powdery mildew. However, information related to genes controlling resistance remains elusive. This study analyzes the virulence structure and the genetic diversity of pathogenic Bgt populations isolated from northeastern (Liaoning, Heilongjiang) and northwestern (Gansu) China, two representative wheat producing areas, on 37 wheat cultivars each carrying a known powdery mildew resistance (Pm) gene. RESULTS: Bgt isolates from northeastern China show higher frequencies of virulence genes than populations from Gansu Province. Many of the known Pm genes failed to provide resistance in this study. However, Pm21 provided 100% resistance to all isolates from all three provinces, obtained during two consecutive years, while Pm13 provided 100% resistance in Gansu. Pm13, Pm16, Pm18, and Pm22 also showed partial resistance in northeastern China, while Pm16, Pm18, Pm22, Pm5 + 6 and Pm2 + 6 +? maintained some resistance in Gansu. Genetic diversity among populations in different regions was detected by cluster analyses using expressed sequence tag-simple sequence repeat (EST-SSR). When the genetic similarity coefficient is relatively high, populations from the same regional origin are mostly clustered into one group while populations from different regions exhibit large genetic differences. CONCLUSION: Pm21 remains the best choice for breeding programs to maintain resistance to Bgt. Only 58% of the isolates tested show a clear correlation between EST-SSR genetic polymorphisms and frequency of virulence gene data.


Assuntos
Ascomicetos/genética , Ascomicetos/patogenicidade , Variação Genética , Ascomicetos/isolamento & purificação , China , Análise por Conglomerados , Etiquetas de Sequências Expressas , Frequência do Gene , Genes Fúngicos , Repetições de Microssatélites/genética , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Triticum/genética , Triticum/microbiologia , Virulência/genética
12.
Plant Dis ; 103(11): 2786-2797, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31535958

RESUMO

Gray blight disease, caused by Pestalotiopsis-like fungi, is one of the deadliest threats to tea (Camellia sinensis) production. However, little information is known about the traits and characteristics of this pathogen. Here, a systematic survey was performed, and a total of 20 representative isolates were obtained from the leaves of tea plants affected by gray blight in two main tea plantations located in Anhui Province, China. Further analyses showed that two isolates were identified as Neopestalotiopsis ellipsospora, three isolates were regarded as Pseudopestalotiopsis chinensis, one isolate was considered as Pseudopestalotiopsis camelliae-sinensis, and the remaining isolates belonged to Pseudopestalotiopsis spp., on the basis of morphological characteristics and multigene phylogenetic analyses of the internal transcribed spacer, ß-tubulin, and translation elongation factor 1-α. Pathogenicity tests indicated that there were significant differences in virulence among the Neopestalotiopsis and Pseudopestalotiopsis isolates when inoculated on the leaves of the tea plant (C. sinensis 'Shuchazao'). Furthermore, varied pathogenicity was also observed for the same isolate when inoculated on different varieties of tea plants. To our knowledge, this is the first record of Neopestalotiopsis ellipsospora and Pseudopestalotiopsis chinensis causing gray blight disease of tea plants in China.


Assuntos
Ascomicetos/classificação , Ascomicetos/patogenicidade , Camellia sinensis , Ascomicetos/genética , Ascomicetos/isolamento & purificação , Camellia sinensis/microbiologia , China , Filogenia , Doenças das Plantas/microbiologia , Virulência
13.
Emerg Infect Dis ; 25(9): 1768-1769, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31441754

RESUMO

Parathyridaria percutanea is an emerging fungus causing subcutaneous phaeohyphomycoses in renal transplant recipients in India. We identified P. percutanea from a patient with subcutaneous phaeohyphomycosis. From our culture collection, we identified the same fungus from 4 similar patients. We found 5 cases previously described in literature.


Assuntos
Ascomicetos/isolamento & purificação , Feoifomicose/diagnóstico , Hipersecreção Hipofisária de ACTH , Adulto , Ascomicetos/genética , Axila , Diagnóstico Diferencial , Antebraço , Humanos , Masculino , Feoifomicose/microbiologia
14.
Int J Syst Evol Microbiol ; 69(12): 3769-3776, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31464661

RESUMO

The genus Verrucoconiothyrium was erected to accommodate Coniothyrium-like species with verruculose conidia. So far, it includes only four species, which have been found in association with plants, and very little is known about their distribution and host preferences. In this study, a Coniothyrium-like fungus isolated from sea water from the north of Portugal was characterised. Phylogenetic analysis, based on sequence data of the internal transcribed spacer and beta-tubulin loci, placed this fungus within the genus Verrucoconiothyrium but clearly distinct from the other known species. A novel species Verrucoconiothyrium ambiguumsp. nov. is described and illustrated. The taxonomic affiliation of the genus Verrucoconiothyrium at the family level was addressed through individual and combined gene genealogies. Our results show that the genus Verrucoconiothyrium is a member of the family Didymellaceae.


Assuntos
Ascomicetos/classificação , Filogenia , Água do Mar/microbiologia , Ascomicetos/isolamento & purificação , DNA Fúngico , Técnicas de Tipagem Micológica , Portugal , Análise de Sequência de DNA , Esporos Fúngicos
15.
World J Microbiol Biotechnol ; 35(7): 108, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31267227

RESUMO

A total of 191 yeasts were isolated from 197 samples collected from eight estuarine mangrove forests along four different coastlines of Thailand (Andaman Sea and the East, North and West coasts of the Gulf of Thailand). Of these, 178 isolates were identified as 32 species in 16 genera of Ascomycota, 12 species in nine genera of Basidiomycota, and 13 isolates as potential new species, respectively. Mangroves located along the Andaman Sea coastline had a higher yeast diversity at the species and genera levels than those along the Gulf of Thailand. Kluyveromyces siamensis was the most frequently isolated species, whilst Candida tropicalis was the only species isolated at all eight sites. Screening isolated yeast strains belonging to genera previously reported as oleaginous yeast plus the 13 potential new species, revealed two oleaginous strains, Rhodotorula sphaerocarpa 11-14.4 and Saitozyma podzolica 11-11.3.1. Both of these strains were isolated from the same mangrove forest on the Andaman Sea coastline. They could accumulate lipid when suspended in glucose solution without any supplementation, while the fatty acid composition and oil profile of Rh. sphaerocarpa 11-14.4 and Sait. podzolica 11-11.3.1 were similar to vegetable oil and cocoa butter, respectively.


Assuntos
Filogenia , Áreas Alagadas , Leveduras/classificação , Leveduras/isolamento & purificação , Ascomicetos/química , Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Basidiomycota/química , Basidiomycota/classificação , Basidiomycota/isolamento & purificação , Biodiversidade , Biocombustíveis , DNA Fúngico/análise , DNA Fúngico/isolamento & purificação , Gorduras na Dieta , Ácidos Graxos/análise , Glucose/metabolismo , Metabolismo dos Lipídeos , Lipídeos/análise , Tipagem Molecular , Óleos Vegetais , RNA Ribossômico/genética , Análise de Sequência , Tailândia , Leveduras/química , Leveduras/genética
16.
World J Microbiol Biotechnol ; 35(7): 105, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31267317

RESUMO

Pseudocercospora fijiensis causes black Sigatoka disease, the most important threat to banana. The cell wall is crucial for fungal biological processes, including pathogenesis. Here, we performed cell wall proteomics analyses of two P. fijiensis strains, the highly virulent Oz2b, and the less virulent C1233 strains. Strains were starved from nitrogen to mimic the host environment. Interestingly, in vitro cultures of the C1233 strain grew faster than Oz2b in PDB medium, suggesting that C1233 survives outside the host better than the highly virulent Oz2b strain. Both strains were submitted to nitrogen starvation and the cell wall proteins were isolated and subjected to nano-HPLC-MS/MS. A total of 2686 proteins were obtained from which only 240 had a known function and thus, bioinformatics analyses were performed on this group. We found that 90 cell wall proteins were shared by both strains, 21 were unique for Oz2b and 39 for C1233. Shared proteins comprised 24 pathogenicity factors, including Avr4 and Ecp6, two effectors from P. fijiensis, while the unique proteins comprised 16 virulence factors in C1233 and 11 in Oz2b. The P. fijiensis cell wall proteome comprised canonical proteins, but thirty percent were atypical, a feature which in other phytopathogens has been interpreted as contamination. However, a comparison with the identities of atypical proteins in other reports suggests that the P. fijiensis proteins we detected were not contaminants. This is the first proteomics analysis of the P. fijiensis cell wall and our results expands the understanding of the fundamental biology of fungal phytopathogens and will help to decipher the molecular mechanisms of pathogenesis and virulence in P. fijiensis.


Assuntos
Ascomicetos/genética , Ascomicetos/metabolismo , Parede Celular/genética , Parede Celular/metabolismo , Proteoma , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Ascomicetos/isolamento & purificação , Ascomicetos/patogenicidade , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Genes Fúngicos/genética , Genoma Fúngico , Musa/microbiologia , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Espectrometria de Massas em Tandem , Virulência
17.
Transpl Infect Dis ; 21(5): e13141, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31283872

RESUMO

BACKGROUND: Scedosporium species and Lomentospora prolificans (S/L) are the second most common causes of invasive mold infections following Aspergillus in lung transplant recipients. METHODS: We assessed the current practices on management of S/L colonization/infection of the lower respiratory tract before and after lung transplantation in a large number of lung transplant centers through an international practice survey from October 2016 to March 2017. RESULTS: A total of 51 respondents from 45 lung transplant centers (17 countries, 4 continents) answered the survey (response rate 58%). S/L colonization was estimated to be detected in candidates by 48% of centers. Only 18% of the centers used a specific medium to detect S/L colonization. Scedosporium spp. colonization was a contraindication to transplantation in 10% of centers whereas L prolificans was a contraindication in 31%; 22% of centers declared having had 1-5 recipients infected with S/L in the past 5 years. CONCLUSIONS: This survey gives an overview of the current practices regarding S/L colonization and infection in lung transplant centers worldwide and underscores the need of S/L culture procedure standardization before implementing prospective studies.


Assuntos
Ascomicetos/isolamento & purificação , Gerenciamento Clínico , Hospedeiro Imunocomprometido , Transplante de Pulmão/efeitos adversos , Micoses/epidemiologia , Scedosporium/isolamento & purificação , Humanos , Internacionalidade , Micoses/etiologia , Estudos Prospectivos , Infecções Respiratórias/etiologia , Infecções Respiratórias/microbiologia , Inquéritos e Questionários
18.
BMC Genomics ; 20(1): 543, 2019 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-31272366

RESUMO

BACKGROUND: Podosphaera xanthii is the main causal agent of powdery mildew disease in cucurbits and is responsible for important yield losses in these crops worldwide. Powdery mildew fungi are obligate biotrophs. In these parasites, biotrophy is determined by the presence of haustoria, which are specialized structures of parasitism developed by these fungi for the acquisition of nutrients and the delivery of effectors. Detailed molecular studies of powdery mildew haustoria are scarce due mainly to difficulties in their isolation. Therefore, their analysis is considered an important challenge for powdery mildew research. The aim of this work was to gain insights into powdery mildew biology by analysing the haustorial transcriptome of P. xanthii. RESULTS: Prior to RNA isolation and massive-scale mRNA sequencing, a flow cytometric approach was developed to isolate P. xanthii haustoria free of visible contaminants. Next, several commercial kits were used to isolate total RNA and to construct the cDNA and Illumina libraries that were finally sequenced by the Illumina NextSeq system. Using this approach, the maximum amount of information from low-quality RNA that could be obtained was used to accomplish the de novo assembly of the P. xanthii haustorial transcriptome. The subsequent analysis of this transcriptome and comparison with the epiphytic transcriptome allowed us to identify the importance of several biological processes for haustorial cells such as protection against reactive oxygen species, the acquisition of different nutrients and genetic regulation mediated by non-coding RNAs. In addition, we could also identify several secreted proteins expressed exclusively in haustoria such as cell adhesion proteins that have not been related to powdery mildew biology to date. CONCLUSIONS: This work provides a novel approach to study the molecular aspects of powdery mildew haustoria. In addition, the results of this study have also allowed us to identify certain previously unknown processes and proteins involved in the biology of powdery mildews that could be essential for their biotrophy and pathogenesis.


Assuntos
Ascomicetos/genética , Ascomicetos/patogenicidade , Proteínas Fúngicas/genética , Ascomicetos/isolamento & purificação , Cucurbita/microbiologia , DNA Complementar , Citometria de Fluxo , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Ontologia Genética , Interações Hospedeiro-Patógeno/genética , Doenças das Plantas/microbiologia , Conformação Proteica , Transcriptoma
19.
Diagn Microbiol Infect Dis ; 95(2): 201-207, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31262546

RESUMO

Chromoblastomycosis (CBM) is a chronic subcutaneous infection caused by melanotic fungi, affecting mainly rural workers in tropical and subtropical regions. Secondary bacterial infections (SBIs) in CBM lesions bring complications to the disease, but little is known about the agents involved. Fungal and bacterial identification and epidemiological profile of 50 patients with CBM were analyzed in this study. Bacteria were tested for susceptibility to antibacterial drugs. Fonseacea pedrosoi and Rhinocladiella aquaspersa were the fungal agents isolated. 88% of the patients presented SBI. Gram-positive bacteria coinfected mainly upper limbs, and Gram-negative bacteria were more isolated from lower limbs. Streptococcus pyogenes and mixed bacterial microbiota were associated with severe lesions. Staphylococcus aureus was associated with mixed infections and consequently with the severity of the infection. Resistance to ß-lactams and methicillin was detected. Our results emphasize the necessity of bacterial culture and susceptibility testing as part of routine monitoring CBM cases.


Assuntos
Cromoblastomicose/microbiologia , Coinfecção/microbiologia , Idoso , Antibacterianos/farmacologia , Ascomicetos/isolamento & purificação , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Brasil/epidemiologia , Cromoblastomicose/diagnóstico , Cromoblastomicose/epidemiologia , Coinfecção/diagnóstico , Coinfecção/epidemiologia , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Microbiota , Pessoa de Meia-Idade , Especificidade da Espécie
20.
J Basic Microbiol ; 59(9): 862-866, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31330054

RESUMO

Multiplex polymerase chain reaction (PCR) is a method for simultaneous identification and detection of multiple pathogenic fungi, however, its complexity limits its application. To simplify the protocol and to improve the effectiveness, three-level designs for six factors (three primers, Taq DNA polymerase, dNTP, Mg2+ ) were constructed to optimize the multiplex PCR system by using the orthogonal design method and the annealing temperature of the PCR reactions was also optimized. Finally, a multiplex PCR system for the simultaneous detection of these three pathogens of maize was successfully established. The reaction volume was 25 µl and the annealing temperature was 57℃. The optimal conditions for multiplex PCR reaction contained 0.48 µmol/L Cl-1/Cl-2, 0.72 µmol/L Bm-1/Bm-2, 0.24 µmol/L Az-1/Az-2, 1.5 U polymerase, 0.35 mmol/L dNTP, and 1.25 mmol/L MgCl2 . The multiplex PCR system can detect Curvularia lunata, Bipolaris maydis, and Aureobasidium zeae in infected plant tissues rapidly with the sensitivity at 10 pg DNA/µl.


Assuntos
Ascomicetos/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Zea mays/microbiologia , Ascomicetos/genética , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia
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