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PubMed1990

Nucleases in the autolysis of filamentous fungi.

RNase and DNase activities were studied in seven fungi of the subdivisions Ascomycotina, Zygomycotina and Basidiomycotina during their autolysis, and extracellular and intracellular RNase and DNase were found. RNase specific activity reached higher levels than DNase specific activity in the culture liquid and mycelial extract, except in Aspergillus nidulans. Generally maximal RNase specific activities were observed at the onset of autolysis in the culture liquid. In the mycelial extract an increase in this activity with the incubation time was observed, except in A. nidulans and Coriolus versicolor. The highest values of DNase specific activities were found at the third day of autolysis in A. nidulans culture liquid and at the thirtieth day of autolysis in Schizophyllum commune mycelial extract. A possible relationship between the culture liquid pH during the autolysis of the studied fungi and the levels of DNase specific activity was observed.

Ascomycota↗
PubMed1991

Gene expression systems for filamentous fungi.

The extraordinary capacity of filamentous fungi to produce large quantities of extracellular protein, together with the advent of DNA-mediated fungal transformation, has resulted in rapid advances in the development of gene expression systems for filamentous fungi. This review focuses on recent developments in the expression of both fungal and non-fungal genes and improvements to the host.

Animals↗
PubMed1992

Evolutionary relationships within the fungi: analyses of nuclear small subunit rRNA sequences.

Nucleotide sequences of the small subunit ribosomal RNA (18S) gene were used to investigate evolutionary relationships within the Fungi. The inferred tree topologies are in general agreement with traditional classifications in the following ways: (1) the Chytridiomycota and Zygomycota appear to be basal groups within the Fungi. (2) The Ascomycota and Basidiomycota are a derived monophyletic group. (3) Relationships within the Ascomycota are concordant with traditional orders and divide the hemi- and euascomycetes into distinct lineages. (4) The Basidiomycota is divided between the holobasidiomycetes and phragmobasidiomycetes. Conflicts with traditional classification were limited to weakly supported branches of the tree. Strongly supported relationships were robust to minor changes in alignment, method of analysis, and various weighting schemes. Weighting, either of transversions or by site, did not convincingly improve the status of poorly supported portions of the tree. The rate of variation at particular sites does not appear to be independent of lineage, suggesting that covariation of sites may be an important phenomenon in these genes.

Animals↗
PubMed2006

Iron uptake in fungi: a system for every source.

Fungi have a remarkable capacity to take up iron when present in any of a wide variety of forms, which include free iron ions, low-affinity iron chelates, siderophore-iron chelates, transferrin, heme, and hemoglobin. Appropriately, these unicellular eukaryotes express a variety of iron uptake systems, some of which are unique to fungi and some of which are present in plants and animals, as well. The reductive system of uptake relies upon the external reduction of ferric salts, chelates, and proteins prior to uptake by a high-affinity, ferrous-specific, oxidase/permease complex. This system recognizes a broad range of substrates. The non-reductive system exhibits specificity for siderophore-iron chelates, and transporters of this system exhibit multiple substrate-dependent intracellular trafficking events.

Cell Wall↗
PubMed2003

Validation of the 'Marburg bone bank system' for thermodisinfection of allogenic femoral head transplants using selected bacteria, fungi, and spores.

The Marburg Bone Bank System 'Lobator sd-2' is widely used to process human femoral heads removed during aseptic surgery by thermal disinfection. The inactivating capacity of the thermodisinfection system was validated in compliance with current standards using a newly developed femoral head model. The following micro-organisms, bacteria and fungi, taken from the American Type Culture Collection were investigated: Staphylococcus aureus, Staphyloccus epidermidis, Enterococcus faecium, Pseudomonas aeruginosa, Bacillus subtilis including spores, Clostridium sporogenes, Mycobacterium terrae, Candida albicans and Aspergillus niger spores. Highly enriched suspensions of these micro-organisms were applied to the centre of the femoral heads. The reduction in the number of micro-organisms was determined by counting the colony-forming units (cfu) before and after processing the spiked test device in the 'Lobator sd-2' system. Vegetative bacteria, fungi and fungal spores were completely inactivated (reduction factor >/=6 log(10)). The numbers of B. subtilis and C. sporogenes spores, both known to be heat-resistant, were reduced by one to two orders of magnitude. These bacteria serve as a model for spore forming pathogens which are not relevant in femoral heads from living donors. By processing human femoral heads from living donors by thermal disinfection using the Marburg Bone Banking system, a high level of safety is achieved regarding clinically relevant pathogens. To further increase the safety of the thermally treated femoral heads, we recommend that the medical history and present state of the donor, as well as the necessary serological tests should be taken into account.

Bacteria↗
PubMed2005

Biosorption of cobalt by fungi from serpentine soil of Andaman.

Fungi belonging to Aspergillus, Mortierella, Paecilomyces, Penicillium, Pythium, Rhizopus and Trichoderma, isolated from serpentine soil of Andaman (India) were screened for cobalt-resistance. Eleven out of total 38 isolated fungi which tolerated > 6.0 mM Co(II) were evaluated for cobalt biosorption using dried mycelial biomass. Maximum Co(II)-loading (1036.5 microM/g, 60 min) was achieved with Mortierella SPS 403 biomass, which removed almost 50% of 4.0 mM cobalt from the aqueous solution. Co(II)-sorption kinetics of Mortierella SPS 403 biomass was fast and appreciable quantities of metal [562.5 microM/g] was adsorbed during first 10 min of incubation. The metal biosorption capacity of the isolate was accelerated with increasing cobalt concentration, while it was reverse with increase of initial biomass. The optimum pH and temperature for Co(II) removal were 7.0 and 30 degrees C, respectively. However, Co(II)-uptake was inhibited in presence of other metals (Pb, Cd, Cu, Ni, Cr and Zn). Freundlich adsorption isotherm appropriately describes Mortierella SPS 403 biomass as an efficient Co(II)-biosorbent.

Adsorption↗
PubMed2006

Metal tolerance and biosorption potential of filamentous fungi isolated from metal contaminated agricultural soil.

Heavy metal analysis of agricultural field soil receiving long-term (>20 years) application of municipal and industrial wastewater showed two- to five-fold accumulation of certain heavy metals as compared to untreated soil. Metal-resistant fungi isolated from wastewater-treated soil belonged to genera Aspergillus, Penicillium, Alternaria, Geotrichum, Fusarium, Rhizopus, Monilia and Trichoderma. Minimum inhibitory concentrations (MIC) for Cd, Ni, Cr, Cu, and Co were determined. The MIC ranged from 0.2 to 5 mg ml(-1) for Cd, followed by Ni (0.1-4 mg ml(-1)), Cr (0.3-7 mg ml(-1)), Cu (0.6-9 mg ml(-1)) and for Co (0.1-5 mg ml(-1)) depending on the isolate. Aspergillus and Rhizopus isolates were tested for their metal biosorption potential for Cr and Cd in vitro. Biosorption experiments were conducted with initial metal concentrations of 2, 4, 6 and 8 mM with a contact time of 4 h and wet fungal biomass (1-5 g) at 25 degrees C. Maximum biosorption of Cr and Cd ions was found at 6 mM initial metal concentration. Aspergillus sp.1 accumulated 1.20 mg of Cr and 2.72 mg of Cd per gram of biomass. Accumulation of these two metals by very tolerant Aspergillus sp.2 isolate was at par with relatively less tolerant Aspergillus sp.1 isolate. Rhizopus sp. accumulated 4.33 mg of Cr and 2.72 mg of Cd per g of biomass. The findings indicated promising biosorption of cadmium and chromium by the Rhizopus and Aspergillus spp. from aqueous solution. There is little, if any, correlation between metal tolerance and biosorption properties of the test fungi.

Adaptation, Physiological↗
PubMed2006

Single-stranded conformation polymorphism of large subunit of ribosomal RNA is best suited to diagnosing fungal infections and differentiating fungi at species level.

In the last decade, various polymerase chain reaction (PCR)-based methods have been developed using ribosomal RNA (rRNA) for the identification of medically important fungi. In the present study, large subunit (LSU) and small subunit (SSU) of fungal rRNA were amplified and analyzed by single-stranded conformation polymorphism (SSCP) of nested PCR, restriction digestion, and SSCP of digested products. The relationship between several clinical isolates of patients suffering from aspergillosis, candidiasis, cryptococcosis, keratitis, and skin and nail infections has been established with standard fungal cultures using the SSU- and LSU-specific primers. Single-stranded conformation polymorphism of restriction profile of amplified products of LSU-specific primers was successfully used to differentiate fungi up to genus and species level.

Classification↗
PubMed2005

Interaction of pathogenic fungi with host cells: Molecular and cellular approaches.

This review provides an overview of several molecular and cellular approaches that are likely to supply insights into the host-fungus interaction. Fungi present intra- and/or extracellular host-parasite interfaces, the parasitism phenomenon being dependent on complementary surface molecules. The entry of the pathogen into the host cell is initiated by the fungus adhering to the cell surface, which generates an uptake signal that may induce its cytoplasmatic internalization. Furthermore, microbial pathogens use a variety of their surface molecules to bind to host extracellular matrix (ECM) components to establish an effective infection. On the other hand, integrins mediate the tight adhesion of cells to the ECM at sites referred to as focal adhesions and also play a role in cell signaling. The phosphorylation process is an important mechanism of cell signaling and regulation; it has been implicated recently in defense strategies against a variety of pathogens that alter host-signaling pathways in order to facilitate their invasion and survival within host cells. The study of signal transduction pathways in virulent fungi is especially important in view of their putative role in the regulation of pathogenicity. This review discusses fungal adherence, changes in cytoskeletal organization and signal transduction in relation to host-fungus interaction.

Cell Adhesion↗
PubMed2005

High diversity of endophytic fungi from the pharmaceutical plant, Heterosmilax japonica Kunth revealed by cultivation-independent approach.

Heterosmilax japonica Kunth is well recognized for its diuretic effects in China. However, little is known about its endophytic fungi. In this study, microbial communities inhabiting the stems of H. japonica in spring and summer were investigated by light microscopy and cultivation-independent approaches, such as RFLP analysis and sequencing of rDNA ITS library. Molecular phylogenetic analysis showed that a broad spectrum of fungi, including Mycosphaerella, Phomopsis, Aureobasidium, Cladosporium, Glomerella, Botryosphaeria, Guignardia, is able to colonize the plants internally. Particularly, several rDNA sequences determined in this study like YJ4-61 are not specifically affiliated with any currently documented fungal sequences in the public database. Several sequence types, such as YJ4-9 and YJ4-70, are significantly similar to some uncultured environmental samples. Furthermore, our result also showed that the samples collected in spring harbored more abundant endophytic populations than that in summer, implying a seasonal fluctuation for the endophytes in H. japonica.

Ascomycota↗
PubMed2003

Septation and cytokinesis in fungi.

Cytokinesis is the ultimate step of a cell cycle resulting in the generation of two progeny. Failure of correct cell division may be lethal for both, mother and daughter cells, and thus such a process must be tightly regulated with other events of the cell cycle. Differing solutions to the same problem have been developed in bacteria and plants while cytokinesis in animal and fungal cells is highly similar and requires a contractile ring containing actomyosin. Cytokinesis in fungi can be viewed as a three-stage process: (i) selection of a division site, (ii) orderly assembly of protein complexes, and finally (iii) dynamic events that lead to a constriction of the contractile ring and septum construction. Elaborate mechanisms known as the Mitotic Exit Network (MEN) and the Septation Initiation Network (SIN) have evolved to link these events, particularly the final steps of cytokinesis, with nuclear division. The purpose of this review was to discuss the latest developments in the fungal field and to describe the central known players required for key steps on the road to cell division. Differences in the cytokinesis of yeast-like fungi that result in complete cell separation in contrast to septation which leads to the compartmentalization of fungal hyphae are highlighted.

Cell Division↗
PubMed2004

Polarity in filamentous fungi: moving beyond the yeast paradigm.

Filamentous fungi grow by the polar extension of hyphae. This polar growth requires the specification of sites of germ tube or branch emergence, followed by the recruitment of the morphogenetic machinery to those sites for localized cell wall deposition. Researchers attempting to understand hyphal morphogenesis have relied upon the powerful paradigm of bud emergence in the yeast Saccharomyces cerevisiae. The yeast paradigm has provided a useful framework, however several features of hyphal morphogenesis, such as the ability to maintain multiple axes of polarity and an extremely rapid extension rate, cannot be explained by simple extrapolation from yeast models. We discuss recent polarity research from filamentous fungi focusing on the position of germ tube emergence, the relaying of positional information via RhoGTPase modules, and the recruitment of morphogenetic machinery components including cytoskeleton, polarisome and ARP2/3 complexes, and the vesicle trafficking system.

Cell Polarity↗
PubMed2004

Different nitrogen sources modulate activity but not expression of glutamine synthetase in arbuscular mycorrhizal fungi.

Glutamine synthetase (GS) is a central enzyme of nitrogen metabolism that allows assimilation of nitrogen and biosynthesis of glutamine. We isolated the cDNA encoding GS from two arbuscular mycorrhizal fungi, Glomus mosseae (GmGln1) and Glomus intraradices (GiGln1). The deduced protein orthologues have a high degree of similarity (92%) with each other as well as with GSs from other fungi. GmGln1 was constitutively expressed during all stages of the fungal life cycle, i.e., spore germination, intraradical and extraradical mycelium. Feeding experiments with different nitrogen sources did not induce any change in the mRNA level of both genes independent of the symbiotic status of the fungus. However, GS activity of extraradical hypahe in G. intraradices was considerably modulated in response to different nitrogen sources. Thus, in a N re-supplementation time-course experiment, GS activity responded quickly to addition of nitrate, ammonium or glutamine. Re-feeding with ammonium produced a general increase in GS activity when compared with hyphae grown in nitrate as a sole N source.

Amino Acid Motifs↗
PubMed2004

Effect of weak acid preservatives on growth of bakery product spoilage fungi at different water activities and pH values.

Inhibition of spoilage organisms from bakery products by weak acid preservatives in concentrations of 0%, 0.003%, 0.03% and 0.3% (w/v) was investigated experimentally on a substrate media with water activity (a(w)) and pH ranging from sourdough-fermented acidic rye bread to alkaline intermediate moisture sponge cake types (a(w) 0.80-0.95, pH 4.7-7.4). Initially, rye bread conditions (a(w) 0.94-0.97 and pH 4.4-4.8) in combination with calcium propionate were investigated. Results showed that the highest concentration of propionate (0.3%) at all conditions apart from high a(w) (0.97) and high pH (4.8) totally inhibited fungal growth for a 2-week period, with the exception of Penicillium roqueforti, Penicillium commune and Eurotium rubrum. Characteristically for the major spoiler of rye bread, P. roqueforti, all three isolates tested were stimulated by propionate and the stimulation was significantly enhanced at high water activity levels. The effect of propionate on production of secondary metabolites (mycophenolic acid, rugulovasine, echinulin, flavoglaucin) was also studied, and variable or isolate dependent results were found. Subsequently, a screening experiment representing a wider range of bakery products was conducted using calcium propionate, potassium sorbate and sodium benzoate. The obtained data was modelled using survival analysis to determine 'spoilage-free time' for the fungi. At the low a(w) level (0.80) only Eurotium species grew within the test period of 30 days. Higher water activity levels as well as higher pH values decreased spoilage-free times of the fungi. The preservative calcium propionate was less effective than potassium sorbate and sodium benzoate.

Acids↗
PubMed2005

Toxigenic fungi isolated from dried vine fruits in Argentina.

To evaluate the potential for mycotoxin production by fungi in dried vine fruits, the mycobiota was determined both before and after surface disinfection. Predominant genera were Aspergillus (50.2%), Eurotium (21.4%) and Penicillium (13.5%). Aspergillus section Nigri ("black aspergilli") were isolated with relatively high frequency. Aspergillus niger was the most common species but only 3 of 293 isolates screened were ochratoxin A (OTA) producers. Aspergillus carbonarius was less common but 96% of 48 strains screened were ochratoxigenic. OTA was not produced by A. japonicus. Other toxigenic fungi detected were A. ochraceus (3 strains produced OTA), Aspergillus flavus (5 strains produced cyclopiazonic acid but not aflatoxins), P. citrinum (19 strains were strong citrinin producers) and Alternaria alternata (15 strains were producers of tenuazonic acid, alternariol and alternariol methyl ether). In spite of the high incidence of A. carbonarius capable of producing OTA, low levels of this toxin were detected in the samples analysed.

Argentina↗
PubMed2026

Herbicolin A, an antifungal lipopeptide produced by Pantoea agglomerans APC 4211 is a promising biocontrol agent against food spoilage fungi.

Fungal contamination of food with yeast and molds is associated with major economic losses due to spoilage and also poses health risks in the form of mycotoxin production. The strain Pantoea agglomerans APC 4211 isolated from leaves of Ilex aquifolium (holly tree) has broad spectrum antifungal activity against a variety of food spoilage fungi. Genomic analysis of the strain confirmed the presence of biosynthetic gene clusters potentially encoding for the enzymatic machinery required for the production of the antifungal lipopeptide herbicolin A. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of the cell-free supernatant (CFS) confirmed the presence of molecular masses corresponding to herbicolin A (1300.8 Da), and herbicolin B (1138 Da). Purified herbicolin A has desirable properties for biotechnological applications, including potent antifungal activity against a range of spoilage fungi, thermal stability and resistance to proteases. The lipopeptide has low cytotoxicity against epithelial cell lines and has minimum inhibitory concentrations (MICs) lower than those of some commercial antifungal drugs (0.2-2.5 mg/L). In a model dairy system (10% skim milk), herbicolin A demonstrated excellent solubility and stability, effectively eliminating Aspergillus niger and Penicillium notatum at a concentration of 5 mg/L. Overall, the study determines herbicolin's A spectrum against food spoilage organisms and examines potential applications in food. In conclusion, herbicolin A is a potent, naturally occurring antifungal agent with the potential to be applied as a biopreservative in food systems, providing a safe, clean-label, and efficient compound for synthetic preservatives replacement.

Pantoea↗
PubMed2004

In vitro studies of endophytic fungi from Tripterygium wilfordii with anti-proliferative activity on human peripheral blood mononuclear cells.

An array of endophytic fungi which habitat in a Chinese medicinal plant, Tripterygium wilfordii Hook. f., (TWHf) were isolated and tested for their suppressive activity on phytohemaglutinin (PHA) stimulated proliferation of human peripheral blood mononuclear cells (PBMC). Out of the 343 isolates, representing 60 different morphotypes, were screened. Amongst the screened fungi, only 15 isolates showed anti-proliferative activity. Ethyl acetate extracts of Pestalotiopsis leucothës, Mucor sp. Verticillium sp. or Pestalotiopsis disseminata, in particular, significantly inhibited the proliferation at doses between 0.12 and 500 microg/ml (stimulation index (S.I.) ranges 0.01-0.70) (P < 0.001-0.05). IC50 values of these four fungal extracts were between 0.75-0.80 +/- 0.12 microg/ml. Trypan blue exclusion test and visual examination of cell morphology demonstrate that the anti-proliferative effect of these extracts was not a sequential consequence of their cytotoxicity. Nevertheless, fungal extracts of Acremonium sp. A and C, Pestalotiopsis suffocata, Morphotype sp. 4 and 5 show a much higher cytotoxic effects on PBMC. Our results indicate that some fungal extracts contain significant amount of immunomodulatory principles.

Adult↗
PubMed2005

Copper complexes of imidazole-2-, pyrrole-2- and indol-3-carbaldehyde thiosemicarbazones: inhibitory activity against fungi and bacteria.

Copper complexes of thiosemicarbazones of imidazole-2-carbaldehyde, pyrrole-2-carbaldehyde and indole-3-carbaldehyde were synthesised and characterised. The antimicrobial properties of the free ligands and their complexes were evaluated against yeasts, moulds and bacteria (Gram-positive and Gram-negative). Some copper chelates exhibited a moderate inhibitory activity, better than that of the corresponding free ligands. In particular, the pyrrole derivative [Cu(HL(2))(2)] proved to be a wide spectrum agent, showing an interesting inhibition of the growth of all Gram-positive bacteria and fungi tested at concentrations of 12-50 microg/mL. In contrast, a selective effect was observed for imidazole and indole chelates against fungi and Gram-positive bacteria, respectively.

Anti-Bacterial Agents↗