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1.
Cell ; 144(4): 539-50, 2011 Feb 18.
Article in English | MEDLINE | ID: mdl-21335236

ABSTRACT

Disassembly of nuclear pore complexes (NPCs) is a decisive event during mitotic entry in cells undergoing open mitosis, yet the molecular mechanisms underlying NPC disassembly are unknown. Using chemical inhibition and depletion experiments we show that NPC disassembly is a phosphorylation-driven process, dependent on CDK1 activity and supported by members of the NIMA-related kinase (Nek) family. We identify phosphorylation of the GLFG-repeat nucleoporin Nup98 as an important step in mitotic NPC disassembly. Mitotic hyperphosphorylation of Nup98 is accomplished by multiple kinases, including CDK1 and Neks. Nuclei carrying a phosphodeficient mutant of Nup98 undergo nuclear envelope breakdown slowly, such that both the dissociation of Nup98 from NPCs and the permeabilization of the nuclear envelope are delayed. Together, our data provide evidence for a phosphorylation-dependent mechanism underlying disintegration of NPCs during prophase. Moreover, we identify mitotic phosphorylation of Nup98 as a rate-limiting step in mitotic NPC disassembly.


Subject(s)
Cell Cycle Proteins/metabolism , Mitosis , Nuclear Pore Complex Proteins/metabolism , Nuclear Pore/metabolism , Protein Serine-Threonine Kinases/metabolism , Animals , Aspergillus/cytology , CDC2 Protein Kinase/metabolism , Cell Cycle , Cell Nucleus/metabolism , HeLa Cells , Humans , Mutation , NIMA-Related Kinase 1 , Nuclear Envelope/metabolism , Phosphorylation , Prophase
2.
Arch Microbiol ; 203(1): 355-366, 2021 Jan.
Article in English | MEDLINE | ID: mdl-32959153

ABSTRACT

Corn, sorghum and wheat grains are used as livestock feed in the world. Identification of black aspergilli associated with these grains is necessary to make sure of the safety of the grains because its occurrence is an indicator of mycotoxin production. Forty-five isolates were isolated from the samples collected from Upper Egypt's markets and identified morphologically based on colony color, conidia, stipe and vesicle size and molecularly by using ß-tubulin and calmodulin genes. Isolates were divided into 30 strains of Aspergillus welwitschiae and 15 strains of A. niger. We have found new criteria in the morphological identification of A. welwitschiae as its colony color was black to brown with yellow edge, but in A. niger was black with white edge, also A. welwitschiae sometimes produced finely-to-distinctly roughened brownish conidia on malt extract agar (MEA) media. Thirteen isolates of A. welwitschiae and six of A. niger were recognized as potential producers for ochratoxin A.


Subject(s)
Aspergillus niger/classification , Aspergillus niger/genetics , Aspergillus/classification , Aspergillus/genetics , Edible Grain/microbiology , Aspergillus/cytology , Aspergillus niger/cytology , Calmodulin/genetics , Mycological Typing Techniques , Ochratoxins , Sorghum/microbiology , Triticum/microbiology , Tubulin/genetics , Zea mays/microbiology
3.
Biotechnol Lett ; 43(1): 61-71, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33026584

ABSTRACT

OBJECTIVE: Evaluation of morphology and secondary metabolites production in Aspergillus terreus ATCC 20542 cultures over a wide range of lactose and yeast extract concentrations from 0.2 up to an extremely high level of 200 g l-l. RESULTS: The morphological differences of mycelial objects were quantified with the use of morphological parameters calculated by applying the tools of digital image analysis. At 200 g l-l of yeast extract clumps and loose hyphae were recorded instead of pellets commonly observed in submerged cultures of A. terreus. Under these conditions the biosynthesis of (+)-geodin and asterric acid was totally blocked, lovastatin formation was found to be at a relatively low level and biomass production turned out to be greater than in the remaining variants, where the pelleted growth was observed. At 200 g l-l of lactose the production of lovastatin, (+)-geodin and asterric acid was visibly stimulated compared to the media containing 0.2, 2 and 20 g l-l of the sugar substrate, but at the same time no traces of butyrolactone I could be detected in the broth. Lactose at the extremely high concentration of 200 g l-l did not induce the drastic morphological changes observed in the case of 200 g l-1 of yeast extract. It was proved that at the C/N values as low as 4 and as high as 374 A. terreus not only continued to display growth but also exhibited the production of secondary metabolites. The use of cultivation media representing the equivalent C/N ratios led to different metabolic and morphological outcomes depending on the concentration of lactose and yeast extract that contributed to the given C/N value. CONCLUSION: The extremely high concentration of yeast extract leads to marked morphological changes of A. terreus and the elimination of (+)-geodin and asterric production, while applying the excess of lactose is stimulatory in terms of lovastatin production.


Subject(s)
Aspergillus , Benzofurans/metabolism , Biological Products/pharmacology , Phenyl Ethers/metabolism , Saccharomyces cerevisiae/chemistry , Aspergillus/cytology , Aspergillus/drug effects , Aspergillus/metabolism , Mycelium/drug effects
4.
Arch Microbiol ; 202(6): 1285-1293, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32128608

ABSTRACT

Mycelial morphogenesis and the production of fungal secretory proteins are still largely unknown. A mutant strain of Aspergillus carbonarius UV-10046 produced abundant polygalacturonase (PG) along with partially saturated canthaxanthin (PSC) at low pH conditions. In the present study, the relationship between PG secretion and PSC biosynthesis was studied using carotenogenic inhibitors and SDS-PAGE electrophoresis. Also the correlation between morphogenesis and PG secretion was investigated by analysing through microscopic studies. From the results, it was observed that secretion of PG was positively influenced by the PSC biosynthesis. The results also showed that the mutant with hairy mycelial structure resulted in higher PG activity when compared to the wild type that lacks hyper branching. From the results, it was confirmed that a mutation might have occurred in the isoprenoid pathway that has helped mutant for survival at acidic conditions. Further, an alteration in the morphogenesis and hyper branching development caused over secretion of PG enzyme in the mutant.


Subject(s)
Aspergillus/enzymology , Aspergillus/genetics , Polygalacturonase/metabolism , Aspergillus/cytology , Canthaxanthin/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism , Morphogenesis , Mutation/genetics
5.
Anal Chem ; 91(19): 12501-12508, 2019 10 01.
Article in English | MEDLINE | ID: mdl-31483612

ABSTRACT

Hyphae of filamentous fungi consist of compartments that are distinct both spatially and functionally, thereby forming a unique multicellular system. Much work has been done mainly using fluorescence imaging to reveal what biomolecules are present in those different hyphal sections and what physiological roles they play. Nevertheless, a holistic understanding of hyphal functions including the polarized growth of hyphae is still lacking because of the difficulty in simultaneous acquisition of spatial and chemical information on various molecular components in living hyphae. Here, we used a multivariate curve resolution-alternating least-squares (MCR-ALS) analysis of Raman hyperspectral imaging data to study in vivo the spatial distributions and chemical properties of major cellular components in the tip, basal, and branching regions of the model fungus Aspergillus nidulans. The MCR-ALS Raman imaging method visualized, without any labeling, the characteristic distributions of cytochromes as well as other components including polysaccharides, noncytochrome proteins, nucleic acids, lipids, and ergosterol in the hyphal regions studied. Furthermore, the intrinsic Raman spectra derived for the first time from the MCR-ALS analysis enabled us to gain otherwise unobtainable chemical insights into those visualized components. We show variations in the relative abundance of cytochromes b and c and in their redox states (reduced vs oxidized form) among the three different representative compartments of A. nidulans hyphae, which could potentially be associated with specific physiological activities and functions of hyphae. The present results demonstrate that our MCR-ALS Raman imaging can serve as a useful tool complementary to the conventional approaches, for elucidating the diverse roles of filamentous fungi at the molecular level.


Subject(s)
Aspergillus/cytology , Cytochromes/metabolism , Molecular Imaging , Spectrum Analysis, Raman , Aryl Hydrocarbon Hydroxylases , Aspergillus/enzymology , Least-Squares Analysis , Multivariate Analysis , Oxidation-Reduction , Steroid Hydroxylases
6.
Med Mycol ; 57(Supplement_2): S155-S160, 2019 Apr 01.
Article in English | MEDLINE | ID: mdl-30816965

ABSTRACT

Aspergillosis is more common among immunocompromised patients with neutropenia or immunosuppression due to corticosteroid use, and infections are typically of the lung or sinuses. For diagnosis, broncholaveolar lavages (BALs) and lung biopsies are the specimens of choice. Culture and microscopic examinations are a must have and laboratory results should immediately be reported to the clinic. Fungal elements (hyphae) display the proof of an infection if present in primarily steril specimens, independent of culture results. Microscopy should be performed preferably using optical brighteners and histopathology using Gomori's methenamine silver stain or Periodic acid-Schiff. Serum and BAL galactomannan assays are recommended as markers for the diagnosis of invasive aspergillosis, PCR should be considered in conjunction with other diagnostic tests. Antifungal treatment decreases GM sensitivity. Pathogen identification to species complex level is strongly recommended for all clinically relevant Aspergillus isolates.


Subject(s)
Aspergillus/isolation & purification , Diagnostic Tests, Routine/methods , Invasive Pulmonary Aspergillosis/diagnosis , Aspergillus/chemistry , Aspergillus/cytology , Aspergillus/genetics , Biopsy , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/microbiology , Galactose/analogs & derivatives , Histocytochemistry , Humans , Lung/microbiology , Lung/pathology , Mannans/analysis , Microbiological Techniques , Microscopy , Polymerase Chain Reaction
7.
Mycopathologia ; 183(2): 423-429, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29128933

ABSTRACT

Invasive fungal disease represents one of the severe complications in haematopoietic stem cell transplant recipients. We describe a case of a patient treated for relapse of chronic lymphoblastic leukaemia 6 years after HSCT. The patient was treated for invasive pulmonary aspergillosis but died 3 months later from multiple organ failures consisting of haemorrhagic necrotizing fungal pneumonia, refractory chronic hepatic graft versus host disease and cytomegalovirus hepatitis. Autopsy samples revealed histopathological evidence of fungal hyphae and an unusual Aspergillus nidulans-like species was isolated in pure culture. More precise identification was achieved by using scanning electron microscopy of ascospores and sequencing of calmodulin gene, and the isolate was subsequently re-identified as A. sublatus (section Nidulantes) and showed good in vitro susceptibility against all classes of antifungals. Commonly used ITS rDNA region and ß-tubulin gene fail to discriminate A. sublatus from related pathogenic species, especially A. quadrilineatus and A. nidulans. Although this is the first case of proven IPA attributed to A. sublatus, we demonstrated that at least some previously reported infections due to A. quadrilineatus were probably caused by this cryptic species.


Subject(s)
Aspergillus/classification , Aspergillus/isolation & purification , Hematopoietic Stem Cell Transplantation/adverse effects , Invasive Pulmonary Aspergillosis/diagnosis , Invasive Pulmonary Aspergillosis/pathology , Leukemia, Lymphocytic, Chronic, B-Cell/complications , Antifungal Agents/administration & dosage , Aspergillus/cytology , Aspergillus/genetics , Calmodulin/genetics , Cluster Analysis , Cytomegalovirus Infections/complications , Cytomegalovirus Infections/diagnosis , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Fatal Outcome , Graft vs Host Disease/complications , Graft vs Host Disease/diagnosis , Hepatitis, Viral, Human/complications , Hepatitis, Viral, Human/diagnosis , Humans , Invasive Pulmonary Aspergillosis/drug therapy , Invasive Pulmonary Aspergillosis/microbiology , Male , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Middle Aged , Phylogeny , Sequence Analysis, DNA , Transplant Recipients , Tubulin/genetics
8.
Environ Microbiol ; 19(2): 687-697, 2017 02.
Article in English | MEDLINE | ID: mdl-27871132

ABSTRACT

Water availability acts as the most stringent constraint for life on Earth. Thus, understanding the water relations of microbial extremophiles is imperative to our ability to increase agricultural productivity (e.g., by enhancing the processing and turnover of dead organic matter in soils of arid regions), reduce human exposure to mycotoxins in buildings and our food-supply chain, prevent the spoilage of foods/animal feeds, books, museum specimens and artworks and better control microbiology of industrial fermentations. Only a small number of microbial systems can retain activity at <0.710 water activity (ISME J 2015 9: 1333-1351). It has long-been considered that the most resilient of these is Xeromyces bisporus, which inhabits sugar-rich substrates (Appl Environ Microbiol 1968 16: 1853-1858). The current study focused on germination of Aspergillus penicillioides, a xerophile which is also able to grow under low humidity and saline conditions. Investigations of germination differed from those reported earlier: firstly, aerially borne conidia were harvested, and then used for inoculations, in their dry condition; secondly, cultures were incubated at 24°C, i.e. below optimum germination temperature, to minimize the possibility of water loss from the substrate; thirdly, cultures remained sealed throughout the 73-day study period (microscopic examination was carried out directly 48 through the Petri plate lid); fourthly, the germination parameters determined were: rates and extent of conidial swelling, production of differentiated germination-structures and septate germlings, and subsequent development of mycelium and/or sporulation; fifthly, assessments were carried out over a range of water-activity values and time points to obtain a complete profile of the germination process. Conidia swelled, formed differentiated germination-structures and then produced septate germlings at a water-activity of just 0.585 (≡58.5% relative humidity), outside the currently understood thermodynamic window for life. Furthermore, analyses of these data suggest a theoretical water-activity minimum of 0.565 for germination of A. penicilliodes. In relation to astrobiology, these findings have an application in understanding the limits to life in extraterrestrial environments. In light of current plans for exploration missions to Mars and other places, and the need to safeguard martian scientific sites and potential resources (including water) for future human habitation, a knowledge-based and effective policy for planetary protection is essential. As it is, Mars-bound spacecraft may frequently be contaminated with aspergilli (including A. penicillioides) and other organisms which, when transported to other planetary bodies, pose a contamination risk. In crafting countermeasures to offset this, it is important to know as precisely as possible the capabilities of these potential interplanetary visitors.


Subject(s)
Aspergillus/growth & development , Spores, Fungal/growth & development , Water/analysis , Aspergillus/cytology , Aspergillus/metabolism , Cell Division , Ecosystem , Exobiology , Extraterrestrial Environment , Humidity , Mycelium/growth & development , Mycelium/metabolism , Temperature , Thermodynamics , Water/metabolism
9.
Microb Pathog ; 113: 135-143, 2017 Dec.
Article in English | MEDLINE | ID: mdl-29074431

ABSTRACT

The present study represents, for the first time, the detailed studies about the hyphal interactions of Aspergillus piperis, as a new antagonist, against some isolated plant pathogenic fungi (Alternaria alternata, Alternaria solani, Botrytis cinerea, Sclerotium cepivorum and Sclerotinia sclerotiorum) in vitro. The bio-controlling capability of A. piperis against the tested phytopathogens was tested using the dual culture method. This experiment revealed that A. piperis had antagonistic activity and reduced the growth of the tested phytopathogens and grew over their mycelia in the paired plates. Also, several antagonistic mechanisms were recorded, in this study, between A. piperis and the tested phytopathogens using the microscopic examination. The bio-controlling activity and the antagonistic mechanisms exhibited by the new antagonist, A. piperis were compared with those obtained by the common antagonist, Trichoderma harzianum against the same phytopathogens. The obtained results showed that, A. piperis was more effective than T. harzianum in inhibiting all the tested species in the dual culture plates. The best result was 81.85% inhibition percentage against S. sclerotiorum by A. piperis while, T. harzianum exhibits only 45.18%. Moreover, several antagonistic mechanisms and hyphal interactions were investigated among the hyphae of both A.piperis and T. harzianum and the hyphae of the tested phytopathogens. These mechanisms were summarized as; mycoparasitism (coiling and penetration of the hyphae) and antibiosis in the form of lysis of the hyphal cells and spores, denaturation and breaking of the hyphae. The indirect interaction (antibiosis) and the direct mycoparasitism were observed by A. piperis against all the tested phytopathogens, but it attacked the hyphae and conidiophores of A. alternata by only the antibiosis interaction. The microscopic examination revealed also that T. harzianum attacked the tested phytopathogens by both antibiosis and mycoparasitism except against A. solani which attacked only by mycoparasitism.


Subject(s)
Antibiosis , Aspergillus/physiology , Hyphae/physiology , Pest Control, Biological , Trichoderma/physiology , Alternaria/isolation & purification , Alternaria/pathogenicity , Ascomycota/isolation & purification , Ascomycota/pathogenicity , Aspergillus/cytology , Aspergillus/growth & development , Botrytis/isolation & purification , Botrytis/physiology , Coculture Techniques , Egypt , Hyphae/cytology , Hyphae/growth & development , Plant Diseases/microbiology , Trichoderma/cytology , Trichoderma/growth & development
10.
J Basic Microbiol ; 57(5): 428-439, 2017 May.
Article in English | MEDLINE | ID: mdl-28370251

ABSTRACT

The infections caused by filamentous fungi are becoming worldwide problem of healthcare systems due to increasing drug-resistance of this microorganism and increasing number of immunocompromised nosocomial patients. These infections are related with Aspergillus ability to form sessile communities referred to as the biofilms. The small compounds known as quorum sensing (QS) molecules allow this microorganism to coordinate all processes taking place during biofilm formation and maturation. In the study presented, the HRMAS 1 H NMR metabolomic approach was applied to define composition of extra and intracellular metabolites produced by biofilmic and planktonic (aka free-swimming) cultures of this microorganism and to evaluate impact of quorum sensing molecule, arachidonic acid (AA) on biofilm formation. The Scanning Electron Microscopy was used to confirm Aspergillus ability to form biofilm in vitro, while multivariate and univariate data analysis was applied to analyze data obtained. The Aspergillus strain was able to form strong biofilm structures in vitro. The statistical analysis revealed significant changes of metabolite production depending on Aspergillus culture type (biofilm vs. plankton), time and presence of QS molecules. The data obtained, if developed, might be used in future NMR diagnostics as markers of Aspergillus biofilm-related infections and lead to shorten time between pathogen identification and introduction of treatment.


Subject(s)
Arachidonic Acid/metabolism , Biofilms/growth & development , Fungi/metabolism , Quorum Sensing/physiology , Aspergillus/cytology , Aspergillus/genetics , Aspergillus/metabolism , Aspergillus/pathogenicity , Cross Infection , Fungi/cytology , Fungi/genetics , Fungi/pathogenicity , Genes, Fungal , Hyphae/cytology , Hyphae/metabolism , Metabolomics/methods , Microscopy, Electron, Scanning , Mycoses/diagnosis , Plankton/physiology
11.
Int J Med Microbiol ; 306(6): 381-90, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27237422

ABSTRACT

Detection of carbohydrate antigens is an important means for diagnosis of invasive fungal infections. For diagnosis of systemic Aspergillus infections, galactomannan is commonly used, the core antigenic structure of which consists of chains of several galactofuranose moieties. In this study, we provide evidence that Fusarium produces at least two distinct galactofuranose antigens: Smaller amounts of galactomannan and larger quantities of a novel antigen recognized by the monoclonal antibody AB135-8. In A. fumigatus, only minor amounts of the AB135-8 antigen are found in supernatants and in the apical regions of hyphae. A galactofuranose-deficient A. fumigatus mutant lacks the AB135-8 antigen, which strongly suggests that galactofuranose is an essential constituent of this antigen. Using a combination of AB135-8 and a galactomannan-specific antibody, we were able to unambiguously differentiate A. fumigatus and Fusarium hyphae in immunohistology. Moreover, since Fusarium releases the AB135-8 antigen, it appears to be a promising target antigen for a serological detection of Fusarium infections.


Subject(s)
Antigens, Fungal/analysis , Aspergillus/isolation & purification , Cell Wall/chemistry , Culture Media/chemistry , Fusarium/isolation & purification , Galactose/analysis , Mycoses/diagnosis , Antibodies, Fungal/immunology , Aspergillus/chemistry , Aspergillus/classification , Aspergillus/cytology , Diagnosis, Differential , Diagnostic Tests, Routine/methods , Fusarium/chemistry , Fusarium/classification , Fusarium/cytology , Immunohistochemistry/methods , Mycoses/microbiology
12.
Bioconjug Chem ; 27(10): 2337-2345, 2016 10 19.
Article in English | MEDLINE | ID: mdl-27623088

ABSTRACT

Self-assembly of nanoparticles on living biotemplate surfaces is a promising route to fabricate nano- or microstructured materials with high efficiency and efficacy. We used filamentous fungi to fabricate microtubules of gold nanoparticles through a novel approach that consists of isolating the hyphal growth from the nanoparticle media. This improved methodology resulted in better morphological control and faster adsorption kinetics, which reduced the time needed to form homogeneous microtubules and allowed for control of microtubule thickness through successive additions of nanoparticles. Differences in the adsorption rates due to modifications in the chemical identity of colloidal gold nanoparticles indicated the influence of secondary metabolites and growth media in the fungi metabolism, which demonstrated the need to choose not only the fungus biotemplate but also the correct medium to obtain microtubules with superior properties.


Subject(s)
Fungi/cytology , Gold/chemistry , Metal Nanoparticles/chemistry , Microtubules , Aspergillus/chemistry , Aspergillus/cytology , Fungi/chemistry , Microscopy, Electron, Scanning Transmission , Microtubules/chemistry , Penicillium/chemistry , Penicillium/cytology , X-Ray Diffraction , Xylariales/chemistry , Xylariales/cytology
13.
J Clin Lab Anal ; 30(5): 745-50, 2016 Sep.
Article in English | MEDLINE | ID: mdl-26926455

ABSTRACT

BACKGROUND: Aspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep-PCR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the identification of Aspergillus strains. MATERIALS AND METHODS: A total of 24 consecutive clinical isolates of Aspergillus were collected during 2012-2014. Conventional morphology and rep-PCR were performed in our Mycology Laboratory. The identification, evaluation, and reporting of strains using MALDI-TOF-MS were performed by BioMérieux Diagnostic, Inc. in Istanbul. DNA sequence analysis of the clinical isolates was performed by the BMLabosis laboratory in Ankara. RESULTS: Samples consisted of 18 (75%) lower respiratory tract specimens, 3 otomycosis (12.5%) ear tissues, 1 sample from keratitis, and 1 sample from a cutaneous wound. According to DNA sequence analysis, 12 (50%) specimens were identified as A. fumigatus, 8 (33.3%) as A. flavus, 3 (12.5%) as A. niger, and 1 (4.2%) as A. terreus. Statistically, there was good agreement between the conventional morphology and rep-PCR and MALDI-TOF methods; kappa values were κ = 0.869, 0.871, and 0.916, respectively (P < 0.001). CONCLUSION: The good level of agreement between the methods included in the present study and sequence method could be due to the identification of Aspergillus strains that were commonly encountered. Therefore, it was concluded that studies conducted with a higher number of isolates, which include other Aspergillus strains, are required.


Subject(s)
Aspergillus/cytology , Aspergillus/isolation & purification , Hospitals, University , Polymerase Chain Reaction/methods , Sequence Analysis, DNA/methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Humans , Turkey
14.
J Invertebr Pathol ; 133: 95-106, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26706117

ABSTRACT

Monotypic stands of common reed and the reed-gall-associated insect assemblages are distributed worldwide. However, fungi associated with these assemblages have not been characterized in detail. Here we examined 5200 individuals (12 species) of immature aculeate hymenopterans or their parasitoids collected at 34 sampling sites in Central Europe. We noticed fungal outgrowth on exoskeletons of 83 (1.60%) larvae and pupae. The most common host was eudominant Pemphredon fabricii. However, the less abundant aculeate hymenopteran reed gall inquilines were infected at higher prevalence, these included Trypoxylon deceptorium, Trypoxylon minus, Hoplitis leucomelana and Hylaeus moricei (all considered new host records). We identified three fungal species, Penicillium buchwaldii (72% of cases), Aspergillus pseudoglaucus (22%) and Penicillium quebecense (6%). When multibrooded nests were affected, only a part of individuals was infected in 62% of cases. The sampling site-specific infection rate reached up to 13%, thus fungal infections should be considered an important variable driving the abundance of gall inquilines. Infections of generalist host species were more frequent than those of reed gall specialists, suggesting that suboptimal conditions decreased the immunocompetence of non-specialized species, which only occasionally nest in reed galls and feed in reed beds.


Subject(s)
Aspergillus/physiology , Hymenoptera/microbiology , Penicillium/physiology , Plant Tumors/microbiology , Animals , Aspergillus/cytology , Aspergillus/genetics , DNA, Fungal/chemistry , Host-Pathogen Interactions , Hymenoptera/classification , Larva/microbiology , Likelihood Functions , Penicillium/cytology , Penicillium/genetics , Phylogeny , Pupa/microbiology
15.
Indian J Exp Biol ; 54(7): 472-476, 2016 Jul.
Article in English | MEDLINE | ID: mdl-29466626

ABSTRACT

Aflatoxins are polyketide secondary metabolites that are produced by certain fungal species in the Aspergillus section Flavi, particularly Aspergillus flavus and Aspergillus parasiticus which contaminate human food as well as animal feed. These are among the most carcinogenic substances known. Due to the toxic and carcinogenic properties of aflatoxins, there is a need to develop reliable methods to detect the presence of aflatoxigenic Aspergilli in contaminated food and feed. Not all Aspergillus strains are able to produce aflatoxins. It requires a detection methodology which can specifically distinguish between the aflatoxin producing and nonproducing strains of Aspergillus. Present communication reports validation of a PCR based detection system based on three genes viz., nor-1, apa-2 and omt-1 involved in aflatoxin biosynthesis, that can specifically distinguish the two aflatoxin producing species viz. Aspergillus flavus ,and Aspergillus parasiticus from non-producers i.e., A. niger, A. fumigates and A. oryzae.


Subject(s)
Aflatoxins/biosynthesis , Aspergillus/pathogenicity , Polymerase Chain Reaction/methods , Aspergillus/cytology , Aspergillus/genetics
16.
J Fluoresc ; 25(4): 803-10, 2015 Jul.
Article in English | MEDLINE | ID: mdl-26123674

ABSTRACT

In this paper, we have described a simple hydrothermal method for preparation of fluorescent carbon dots (C-dots) using Carica papaya juice as a precursor. The synthesized C-dots show emission peak at 461 nm with a quantum yield of 7.0 %. The biocompatible nature of C-dots was confirmed by a cytotoxicity assay on E. coli. The C-dots were used as fluorescent probes for imaging of bacterial (Bacillus subtilis) and fungal (Aspergillus aculeatus) cells and emitted green and red colors under different excitation wavelengths, which indicates that the C-dots can be used as a promising material for cell imaging.


Subject(s)
Aspergillus/cytology , Bacillus subtilis/cytology , Beverages , Carbon/chemistry , Carica/chemistry , Fluorescent Dyes/chemistry , Molecular Imaging/methods , Quantum Dots , Image Processing, Computer-Assisted , Microbial Viability , Nitrogen/chemistry
17.
Biosci Biotechnol Biochem ; 79(3): 488-95, 2015.
Article in English | MEDLINE | ID: mdl-25410617

ABSTRACT

ClbR is a Zn(II)2Cys6 transcriptional activator that controls the expression of cellulase-related genes in response to Avicel and cellobiose in Aspergillus aculeatus. A clbR-overexpressing strain (clbR-OE) that expresses the clbR gene at levels sevenfold higher than the control strain sustainably produced xylanolytic and cellulolytic activities during 10-day cultivation of A. aculeatus, enabling synchronization of xylanolytic and cellulolytic activities at a maximum level. However, clbR overexpression did not simultaneously increase levels of all xylanolytic and cellulolytic enzymes. Peptide mass fingerprint analysis revealed markedly increased production of FIa-xylanase in clbR-OE, whereas expression of FIII-avicelase and FII-carboxymethyl cellulase was unaffected and expression of hydrocellulase was lower in clbR-OE than in the control. Northern blot analysis confirmed that these effects of clbR overexpression on enzyme production were mediated at the transcriptional level. These data suggest that ClbR participates in diverse signaling pathways to control the expression of cellulosic biomass-degrading enzymes in A. aculeatus.


Subject(s)
Aspergillus/enzymology , Aspergillus/genetics , Biomass , Cellulase/biosynthesis , Cellulose/metabolism , Endo-1,4-beta Xylanases/biosynthesis , Transcription Factors/genetics , Aspergillus/cytology , Aspergillus/metabolism , Cellulase/metabolism , Endo-1,4-beta Xylanases/metabolism , Gene Expression , Signal Transduction/genetics
18.
BMC Genomics ; 15: 1011, 2014 Nov 22.
Article in English | MEDLINE | ID: mdl-25416206

ABSTRACT

BACKGROUND: Chemical mutagenesis screens are useful to identify mutants involved in biological processes of interest. Identifying the mutation from such screens, however, often fails when using methodologies involving transformation of the mutant to wild type phenotype with DNA libraries. RESULTS: Here we analyzed Illumina sequence of a chemically derived mutant of Aspergillus nidulans and identified a gene encoding a C2H2 transcription factor termed RsrA for regulator of stress response. RsrA is conserved in filamentous fungal genomes, and upon deleting the gene in three Aspergillus species (A. nidulans, A. flavus and A. fumigatus), we found two conserved phenotypes: enhanced resistance to oxidative stress and reduction in sporulation processes. For all species, rsrA deletion mutants were more resistant to hydrogen peroxide treatment. In depth examination of this latter characteristic in A. nidulans showed that upon exposure to hydrogen peroxide, RsrA loss resulted in global up-regulation of several components of the oxidative stress metabolome including the expression of napA and atfA, the two bZIP transcription factors mediating resistance to reactive oxygen species (ROS) as well as NapA targets in thioredoxin and glutathione systems. Coupling transcriptional data with examination of ΔrsrAΔatfA and ΔrsrAΔnapA double mutants indicate that RsrA primarily operates through NapA-mediated stress response pathways. A model of RsrA regulation of ROS response in Aspergillus is presented. CONCLUSION: RsrA, found in a highly syntenic region in Aspergillus genomes, coordinates a NapA mediated oxidative response in Aspergillus fungi.


Subject(s)
Aspergillus/genetics , Conserved Sequence , Fungal Proteins/metabolism , Oxidative Stress , Sequence Analysis, DNA , Signal Transduction , Transcription Factors/genetics , Aspergillus/cytology , Aspergillus/drug effects , Blotting, Southern , Chromatography, Thin Layer , Fungal Proteins/genetics , Gene Deletion , Gene Expression Regulation, Fungal/drug effects , Genetic Complementation Test , Hydrogen Peroxide/pharmacology , Meiosis/drug effects , Mitosis/drug effects , Models, Biological , Oxidative Stress/drug effects , Oxidative Stress/genetics , Phenotype , Reproduction/drug effects , Signal Transduction/drug effects , Signal Transduction/genetics , Spores, Fungal/drug effects , Spores, Fungal/growth & development , Sterigmatocystin/biosynthesis , Synteny/genetics , Transcription Factors/metabolism , Transcription, Genetic/drug effects
19.
Eukaryot Cell ; 12(7): 962-9, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23650087

ABSTRACT

Aspergillus lentulus was described in 2005 as a new species within the A. fumigatus sensu lato complex. It is an opportunistic human pathogen causing invasive aspergillosis with high mortality rates, and it has been isolated from clinical and environmental sources. The species is morphologically nearly identical to A. fumigatus sensu stricto, and this similarity has resulted in their frequent misidentification. Comparative studies show that A. lentulus has some distinguishing growth features and decreased in vitro susceptibility to several antifungal agents, including amphotericin B and caspofungin. Similar to the once-presumed-asexual A. fumigatus, it has only been known to reproduce mitotically. However, we now show that A. lentulus has a heterothallic sexual breeding system. A PCR-based mating-type diagnostic detected isolates of either the MAT1-1 or MAT1-2 genotype, and examination of 26 worldwide clinical and environmental isolates revealed similar ratios of the two mating types (38% versus 62%, respectively). MAT1-1 and MAT1-2 idiomorph regions were analyzed, revealing the presence of characteristic alpha and high-mobility-group (HMG) domain genes, together with other more unusual features such as a MAT1-2-4 gene. We then demonstrated that A. lentulus possesses a functional sexual cycle with mature cleistothecia, containing heat-resistant ascospores, being produced after 3 weeks of incubation. Recombination was confirmed using molecular markers. However, isolates of A. lentulus failed to cross with highly fertile strains of A. fumigatus, demonstrating reproductive isolation between these sibling species. The discovery of the A. lentulus sexual stage has significant implications for the management of drug resistance and control of invasive aspergillosis associated with this emerging fungal pathogen.


Subject(s)
Aspergillus fumigatus/physiology , Aspergillus/physiology , Aspergillus/cytology , Aspergillus/genetics , Aspergillus/isolation & purification , Crosses, Genetic , Genes, Mating Type, Fungal , Genetic Loci/genetics , Genotype , Humans , Meiosis/genetics , Recombination, Genetic/genetics , Reproduction , Spores, Fungal/ultrastructure
20.
Mycoses ; 56(4): 477-81, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23448599

ABSTRACT

The incidence of onychomycosis due to non-dermatophyte moulds (NDM) is increasing. Aspergillus terreus is relatively undocumented as an agent of this fungal infection. The aim of this work is to show the prevalence of onychomycosis caused by A. terreus and to describe its clinical features. Nail samples were collected for microscopic examination and culturing in selective media. All cases of onychomycosis due to NDM were confirmed by a second sample. Aspergillus terreus isolates were identified through their morphological characteristics and using molecular methods. A total of 2485 samples were obtained. Positive cultures were obtained in 1639 samples. From 124 NDM confirmed cultures, 23 were identified as A. terreus (18.5%). Superficial white onychomycosis was the most frequent clinical pattern. A high percentage was found in fingernails. The prevalence of A. terreus in this study considerably exceeded the percentages reported by other authors. Onychomycosis due to A. terreus presents similar clinical patterns to those caused by dermatophytes, but is difficult to eradicate and is associated with less predictable treatment outcomes. Better knowledge of the aetiology of A. terreus may be important for accomplishing more accurate and effective treatment.


Subject(s)
Aspergillus/classification , Aspergillus/isolation & purification , Onychomycosis/epidemiology , Onychomycosis/microbiology , Adult , Aged , Aspergillus/cytology , Aspergillus/genetics , Humans , Microbiological Techniques , Microscopy , Middle Aged , Molecular Diagnostic Techniques , Nails/microbiology , Nails/pathology , Onychomycosis/pathology , Prevalence
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