During our investigation on the endophytic fungi of Azadirachta indica, the strain YM 311593 was obtained from the fruit of the plant. The culture extract of the strain showed antifungal activities against four phytopathogenic fungi. Based on the morphological features and phylogenetic definition, the strain YM 311593 was identified as Paraconiothyrium sp. Four xanthones and one anthraquinone were obtained from the extract of the fermentation broth of the strain. They were characterized to be globosuxanthone A (1), vertixanthone (2), hydroxyvertixanthone (3), 3,8-dihydroxy-1-methy1-9H- xanthen-9-one (4), and danthron (5), respectively, by spectroscopic elucidation. Furthermore, the absolute configuration of 1 was deduced by X-ray diffraction analysis. Besides, compound 4 was firstly found from natural sources. The antifungal activities of compounds 1-5 towards four phytopathogens were assayed using broth microdilution method. Among them, globosuxanthone A (1) showed obvious antifungal activity towards Fusarium graminearum, Fusarium solani, and Botrytis cinerea with MIC values of 4, 8, and 16 μg/mL, respectively.
- MeSH
- Ascomycota chemie izolace a purifikace MeSH
- Azadirachta mikrobiologie MeSH
- endofyty chemie izolace a purifikace MeSH
- fungicidy průmyslové izolace a purifikace farmakologie MeSH
- fylogeneze MeSH
- houby účinky léků MeSH
- kultivační média chemie MeSH
- ovoce mikrobiologie MeSH
- xantony izolace a purifikace farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
Chromoblastomykózy a feohyfomykózy jsou méně časté mykotické infekce, které jsou vyvolávány tmavě pigmentovanými houbami. Faktory virulence hrají významnou úlohu v patogenezi těchto onemocnění. Muriformní buňky jsou jedním z těchto faktorů a jsou také nejdůležitějším prvkem pro diferenciální diagnostiku chromoblastomykózy a feohyfomykózy z klinického materiálu pomocí různých technik barvení mikroskopických preparátů. Přesná identifikace původců chromoblastomykóz a feohyfomykóz je velmi důležitá z hlediska správného a včasného přístupu k antimykotické terapii, a proto je vhodné využít pro konfirmaci druhové identifikace etiologického agens sekvenaci DNA z narostlé kultury. Včasná diagnostika může být klíčová, zejména v případě invazivních forem těchto infekcí. Některé imunochemické metody mohou být při navádění diagnostika správným směrem nápomocné a detekce DNA metodou polymerázové řetězové reakce přímo z klinického materiálu se zdá být užitečná pro identifikaci původců těchto závažných a život ohrožujících infekcí.
Chromoblastomycosis and phaeohyphomycosis are less common fungal infections caused by dark-pigmented fungi. Virulence factors play an important role in the pathogenesis of these diseases. One of these factors, muriform cells, are the most important element for differential diagnosis of chromoblastomycosis and phaeohyphomycosis using clinical samples and various staining techniques. Accurate identification of pathogens causing chromoblastomycosis and phaeohyphomycosis is very important for correct and early antifungal therapy. Therefore, species identification of the etiological agent should be confirmed by sequencing of DNA from the culture. Early diagnosis may be crucial, especially in case of invasive forms of these infections. The diagnosis may be guided by some immunohistochemistry methods and DNA detection using polymerase chain reaction directly from clinical samples seems to be useful for identification of pathogens causing these severe and life-threatening infections.
- MeSH
- Ascomycota enzymologie izolace a purifikace patogenita MeSH
- barvení a značení metody MeSH
- chromoblastomykóza * mikrobiologie patologie MeSH
- faktory virulence analýza imunologie izolace a purifikace MeSH
- histologické techniky metody MeSH
- imunohistochemie metody MeSH
- interakce hostitele a patogenu genetika imunologie MeSH
- kultivační média speciální MeSH
- lidé MeSH
- phaeohyphomykóza * mikrobiologie patologie MeSH
- polymerázová řetězová reakce metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
The psychrophilic (cold-loving) fungus Pseudogymnoascus destructans was discovered more than a decade ago to be the pathogen responsible for white-nose syndrome, an emerging disease of North American bats causing unprecedented population declines. The same species of fungus is found in Europe but without associated mortality in bats. We found P. destructans was infected with a mycovirus [named Pseudogymnoascus destructans partitivirus 1 (PdPV-1)]. The virus is bipartite, containing two double-stranded RNA (dsRNA) segments designated as dsRNA1 and dsRNA2. The cDNA sequences revealed that dsRNA1 dsRNA is 1,683 bp in length with an open reading frame (ORF) that encodes 539 amino acids (molecular mass of 62.7 kDa); dsRNA2 dsRNA is 1,524 bp in length with an ORF that encodes 434 amino acids (molecular mass of 46.9 kDa). The dsRNA1 ORF contains motifs representative of RNA-dependent RNA polymerase (RdRp), whereas the dsRNA2 ORF sequence showed homology with the putative capsid proteins (CPs) of mycoviruses. Phylogenetic analyses with PdPV-1 RdRp and CP sequences indicated that both segments constitute the genome of a novel virus in the family Partitiviridae. The purified virions were isometric with an estimated diameter of 33 nm. Reverse transcription PCR (RT-PCR) and sequencing revealed that all US isolates and a subset of Czech Republic isolates of P. destructans were infected with PdPV-1. However, PdPV-1 appears to be not widely dispersed in the fungal genus Pseudogymnoascus, as non-pathogenic fungi P. appendiculatus (1 isolate) and P. roseus (6 isolates) tested negative. P. destructans PdPV-1 could be a valuable tool to investigate fungal biogeography and the host-pathogen interactions in bat WNS.
- MeSH
- Ascomycota izolace a purifikace virologie MeSH
- Chiroptera mikrobiologie MeSH
- dvouvláknová RNA genetika izolace a purifikace MeSH
- fylogeneze MeSH
- mykoviry genetika fyziologie ultrastruktura MeSH
- RNA virová genetika izolace a purifikace MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- virové proteiny chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Geografické názvy
- Česká republika MeSH
- Spojené státy americké MeSH
Bradymyces oncorhynchi is a poorly known melanised fungal species that has been isolated only from a hyperaemic focus near the enlarged spleen of a rainbow trout. Although the pathogenicity of this species to fish is suspected, it has not been fully confirmed. Four laboratory experiments were conducted to test the effect of the fungus on the health of rainbow trout fingerlings. Mycelia were cultivated under different conditions to increase inoculum variability and to test the impact of the conditions on the pathogenicity. The inoculum was subsequently administered by the intraperitoneal route. The clinical manifestations, gross pathological lesions and histopathological changes were identical in all experimental groups, i.e. lethargy, inappetence, anorexia, weight reduction, pale gills, full-thickness ulceration of the abdominal wall, muscle atrophy, haemorrhagic or pale liver, fat-altered pyloric region with haemorrhages, enlarged spleen, haemorrhagic ascites, systemic granulomatous lesions with the presence of melanised hyphae (phaeohyphomycosis), multifocal granulomatous hepatitis, perivascular lymphocytic infiltration, vacuolation of hepatocytes and vacuolation of the kidney tubule epithelium. No statistically significant differences were found in the survival of the fish with respect to experimental settings. The survival period ranged between 3 and 135 days.
Medicinal plants have been studied for potential endophytic interactions and numerous studies have provided evidence that seeds harbor diverse microbial communities, not only on their surfaces but also within the embryo. Adenosine deaminase (ADA) is known as a potential therapeutic target for the treatment of lymphoproliferative disorders and cancer. Therefore, in this study, 20 types of medicinal plant seeds were used to screen endophytic fungi with tissue homogenate and streak. In addition, 128 morphologically distinct endophyte strains were isolated and their ADA inhibitory activity determined by a spectrophotometric assay. The strain with the highest inhibitory activity was identified as Cochliobolus sp. Seven compounds were isolated from the strain using a chromatography method. Compound 3 showed the highest ADA inhibitory activity and was identified as 5-hydroxy-2-hydroxymethyl-4H-pyran-4-one, based on the results of 1H and 13C NMR spectroscopy. The results of molecular docking suggested that compound 3 binds to the active site and the nonspecific binding site of the ADA. Furthermore, we found that compound 3 is a mixed ADA inhibitor. These results indicate that endophytic strains are a promising source of ADA inhibitors and that compound 3 may be a superior source for use in the preparation of biologically active ADA inhibitor compounds used to treat cancer.
- MeSH
- adenosindeaminasa chemie metabolismus MeSH
- Ascomycota chemie klasifikace genetika izolace a purifikace MeSH
- endofyty chemie klasifikace genetika izolace a purifikace MeSH
- inhibitory adenosindeaminasy chemie farmakologie MeSH
- léčivé rostliny mikrobiologie MeSH
- lidé MeSH
- magnetická rezonanční spektroskopie MeSH
- nádory farmakoterapie enzymologie MeSH
- semena rostlinná mikrobiologie MeSH
- simulace molekulového dockingu MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
This is the first exhaustive report on the fungal community biodiversity in hypersaline water in România. A total of 27 fungal strains (19 molds and eight yeast) have been isolated from Lopătari hypersaline water, Buzau County. Based on classical investigation, these strains have been identified as belonging to the genera Aureobasidium, Alternaria, Aspergillus, Penicillium, and Fusarium. The molecular characterization of fungal isolates at species level was performed using PCR-RFLP analysis of the 5.8S-ITS region. PCR products were digested with different combinations of endonucleases. The most frequently isolated species were Aspergillus niger (14.81% of all isolates), A. versicolor, (14.81%) and Penicillium crustosum (14.81%). In addition, ribosomal restriction patterns which exhibited profiles specific to Aureobasidium pullulans were derived, and to discriminate between Aureobasidium isolates, the elongase-encoding gene (ELO) was chosen as a genetic marker followed by digestion with endonuclease HhaI. Five yeast isolates displayed restriction patterns corresponding to Aureobasidium melanogenum (18.52%) and three isolates to Aureobasidium pullulans (11.11%). In addition, the RFLP types of Aureobasidium pullulans varieties with HhaI are clearly distinguished and could be applied to assess the intraspecific variability.
- MeSH
- Ascomycota genetika izolace a purifikace metabolismus MeSH
- Aspergillus genetika izolace a purifikace metabolismus MeSH
- biodiverzita MeSH
- DNA fungální genetika MeSH
- fylogeneze MeSH
- houby klasifikace genetika izolace a purifikace MeSH
- kvasinky genetika izolace a purifikace metabolismus MeSH
- mikrobiologie vody * MeSH
- polymerázová řetězová reakce MeSH
- polymorfismus délky restrikčních fragmentů * MeSH
- řeky chemie mikrobiologie MeSH
- ribozomální DNA genetika MeSH
- sekvenční analýza DNA MeSH
- slané vody * MeSH
- tolerance k soli * MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Rumunsko MeSH
Cerebral abscesses caused by dark-pigmented Fonsecaea fungi are rare, especially in otherwise healthy individuals. In this case report, we present a 61-year-old man from Moldova, living in the Czech Republic, who had worked as a locksmith on oil platforms in Turkmenistan, Kazakhstan, Sudan, and Iraq since 1999, and was admitted to a neurology ward for a sudden motion disorder of the right leg, dysarthria, and hypomimia. Imaging revealed presence of expansive focus around the left lateral ventricle of the brain and a pronounced peripheral edema. The intracranial infectious focus was excised under intraoperative SonoWand guidance. Tissue samples were histologically positive for dark-pigmented hyphae, suggesting dematiaceous fungi. Therefore, liposomal amphotericin B therapy was initiated immediately. Fonsecaea monophora was provisionally identified using ITS rDNA region sequencing directly from brain tissue. The identification was subsequently confirmed by cultivation and DNA sequencing from culture. The strain exhibited in vitro sensitive to voriconazole (MIC = 0.016 μg/mL) and resistance to amphotericin B (MIC = 4 μg/mL); therefore, the amphotericin B was replaced with voriconazole. Postoperatively, a significant clinical improvement was observed and no additional surgery was required. Based on the literature review, this is the third documented case of cerebral infection due to this pathogen in patients without underlying conditions and the first such case in Europe.
- MeSH
- absces mozku diagnostické zobrazování mikrobiologie chirurgie MeSH
- amfotericin B terapeutické užití MeSH
- antifungální látky farmakologie terapeutické užití MeSH
- Ascomycota účinky léků genetika izolace a purifikace MeSH
- imunokompetence MeSH
- lidé středního věku MeSH
- lidé MeSH
- mykózy diagnóza diagnostické zobrazování MeSH
- ribozomální DNA genetika MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- přehledy MeSH
- Geografické názvy
- Česká republika MeSH
Neoscytalidium (or N.) dimidiatum and N. novaehollandiae are two aggressive plant pathogenic species that affect several agricultural crops. Early detection and identification of these fungi are of critical importance to bring about the effective minimization to the threat they pose to the infected plants. Herein, two species of Neoscytalidium were rapidly discriminated by utilizing the rRNA internal transcribed (ITS4-5.8S-ITS5) PCR primers. A total of 100 isolates of Neoscytalidium species, which were isolated from Iraqi canker-infected fig trees, were included in this study. Two discrete electrophoretic PCR bands were observed in Neoscytalidium isolates-A-variants were about 546 bp, while B-variants were about 993 bp in length. The comprehensive phylogenetic analysis of both DNA variants revealed that A-variants resided between N. novaehollandiae and N. hyalinum, while B-variants were closely related to N. dimidiatum. Furthermore, the highly specific re-constructed tree of both electrophoretic variants demonstrated that B-variants share a high similarity with N. novaehollandiae. Additionally, the secondary structures for both variants were predicted computationally to reveal the structural patterns that each variant follows. In conclusion, a small rRNA locus comprising 22 nucleotides that differs in the two variants is potentially responsible for this species-specific classification. The main divergence in the amplified loci led to the classification of these fungal variants into two main species, namely N. dimidiatum and N. novaehollandiae, demonstrating that the amplification by ITS4-ITS5 rRNA fragment is a beneficial strategy that can be employed for the assessment of Neoscytalidium diversity in the natural ecosystems.
- MeSH
- Ascomycota klasifikace genetika izolace a purifikace MeSH
- DNA fungální genetika MeSH
- druhová specificita MeSH
- elektroforéza v agarovém gelu * MeSH
- Ficus mikrobiologie MeSH
- fylogeneze MeSH
- genetická variace * MeSH
- mezerníky ribozomální DNA chemie genetika MeSH
- nemoci rostlin mikrobiologie MeSH
- polymerázová řetězová reakce MeSH
- RNA ribozomální 5.8S chemie genetika MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
In field and laboratory experiments during 2014-2017, we investigated the influence of lower and higher cultivation intensity of wheat and ecological factors (weather-temperature and rainfalls, year) on the occurrence of phytopathogenic fungi on the leaves of winter wheat. The prevailing fungi in those years were Mycosphaerella graminicola (Fuckel) J. Schrott and Pyrenophora tritici-repentis (Died.) Drechsler. Using cluster analysis, we statistically evaluated interrelationships of known factors on the abundance of the fungi on leaf surfaces. Our results showed strongest correlation with Mycosphaerella graminicola and Pyrenophora tritici-repentis abundance to be with lower cultivation intensity and year done by the temperature and the rainfalls. The two pathogens-Puccinia tritici Oerst and Hymenula cerealis Ellis & Everh. occurred only very sparsely in some years and had little positive or negative correlation with named factors. The semi-early and semi-late winter wheat varieties Matchball, Annie, Fakir, and Tobak were used for our experiments. Higher cultivation intensity had protective effect against leaf phytopathogenic fungi.
Rock-inhabiting fungi (RIF) are adapted to thrive in oligotrophic environments and to survive under conditions of abiotic stress. Under these circumstances, they form biocoenoses with other tolerant organisms, such as lichens, or with less specific phototrophic consortia of aerial algae or cyanobacteria. RIF are phylogenetically diverse, and their plastic morphological characters hamper the straightforward species delimitation of many taxa. Here, we present a phylogenetic study of two RIF genera, Lichenothelia and Saxomyces. Representatives of both genera inhabit rather similar niches on rocks, but their phylogenetic relationships are unknown so far. The cosmopolitan genus Lichenothelia is recognized by characters of fertile ascomata and includes species with different life strategies. In contrast, Saxomyces species were described exclusively by mycelial characters found in cultured isolates from rock samples collected at high alpine elevations. Here, we use an extended taxon sampling of Dothideomycetes to study the phylogenetic relationships of both Lichenothelia and Saxomyces. We consider environmental samples, type species, and cultured isolates of both genera and demonstrate their paraphyly, as well as the occurrence of teleomorphs in Saxomyces. We applied three species delimitation methods to improve species recognition based on molecular data. We show the distinctiveness of the two main lineages of Lichenothelia (Lichenotheliales s. str.) and Saxomyces and discuss differences in species delimitation depending on molecular markers or methods. We revise the taxonomy of the two genera and describe three new taxa, Lichenothelia papilliformis, L. muriformis, and Saxomyces americanus, and the teleomorph of S. penninicus.
