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Positions of multiple insertions in SSU rDNA of lichen-forming fungi.

Lichen-forming fungi, in symbiotic associations with algae, frequently have nuclear small subunit ribosomal DNA (SSU rDNA) longer than the 1,800 nucleotides typical for eukaryotes. The lichen-forming ascomycetous fungus Lecanora dispersa contains insertions at eight distinct positions of its SSU rDNA; the lichen-forming fungi Calicium tricolor and Porpidia crustulata each contain one insertion. Insertions are not limited to fungi that form lichens; the lichen ally Mycocalicium albonigrum also contains two insertions. Of the 11 insertion positions now reported for lichen-forming fungi and this ally, 6 positions are known only from lichen-forming fungi. Including the 4 newly reported in this study, insertions are now known from at least 17 positions among all reported SSU rDNA sequences. Insertions, most of which are Group I introns, are reported in fungal and protistan lineages and occur at corresponding positions in genomes as phylogenetically distant as the nuclei of fungi, green algae, and red algae. Many of these positions are exposed in the mature rRNA tertiary structure and may be subject to independent insertion of introns. Insertion of introns, accompanied by their sporadic loss, accounts for the scattered distribution of insertions observed within the SSU rDNA of these diverse organisms.

Base Sequence↗

Distribution of anaerobic fungi in the digestive tract of cattle and their survival in faeces.

A most probable numbers procedure was used to enumerate populations of anaerobic fungi in the digesta and faeces of cattle. Anaerobic fungi were isolated from the rumen, omasum, abomasum, small intestine, caecum, large intestine and faeces. By determining the amount of digesta in each organ of the digestive tract, it was possible to estimate the total population of anaerobic fungi in cattle and make comparisons between populations in different organs. In addition to enumerating anaerobic fungi in freshly collected samples, they were quantified in digesta and faeces which had been dried at ambient temperature and stored in air for up to 9 months. These experiments showed that a higher proportion of the anaerobic fungi present in the hindgut and faeces were able to withstand desiccation than those present within the gastric and pre-gastric organs. Our results support the hypothesis that the life cycle of anaerobic fungi consists of three stages; the motile zoospore, the vegetative thallus and an aero-tolerant survival stage (cyst or resistant zoosporangium).

Anaerobiosis↗

Studies on shared antigenic/allergenic components among fungi.

Fungal allergens have been found to be one of the most prevalent aeroallergens in India. Knowledge of shared/unique components among different fungi is necessary for proper diagnosis and treatment of patients allergic to fungi. In the present study, crude extracts (CE) of 11 common fungi (Alternaria alternata, Aspergillus flavus, Asp. fumigatus, Asp. niger, Asp. tamarii, Asp. versicolor, Cladosporium herbarum, Curvularia lunata, Mucor hiemalis, Penicillium citrinum, and Fusarium solani) were characterized by isoelectric focusing (IEF), SDS-PAGE, and immunoblot. On IEF (pI 3-9), the number of protein bands was found to be greatest (46) in M. hiemalis extract. SDS-PAGE exhibited a varied number of bands, generally 18-40, with mol. mass ranging from 14 to 100 kDa. IgG-specific immunoprint using rabbit anti-F. solani CF antibodies demonstrated a mol. mass distribution of shared antigenic proteins of 14-100 kDa in most of the fungi. Shared allergenicity was observed in a number of allergenic proteins in fungal extracts with mol. mass ranging between 14 and 70 kDa on IgE-specific immunoblot using pooled sera of patients allergic to Fusarium. A 45-kDa protein was found to be common among these fungi on immunoblot with patients as well as with rabbit antibodies. F. solani CF extract contained more antigenic/allergenic proteins than F. solani CE. It was concluded that F. solani CF shared several antigenic/allergenic components with CE of other common fungi. This fact needs to be taken into account when fungal extracts are used in diagnosis and immunotherapy of allergic patients.

Adolescent↗

In vitro suppression of fungi caused by combinations of apparently non-antagonistic soil bacteria.

We hypothesized that apparently non-antagonistic soil bacteria may contribute to suppression of fungi during competitive interactions with other bacteria. Four soil bacteria (Brevundimonas sp., Luteibacter sp., Pedobacter sp. and Pseudomonas sp.) that exhibited little or no visible antifungal activity on different agar media were prescribed. Single and mixed strains of these species were tested for antagonism on a nutrient-poor agar medium against the plant pathogenic fungi Fusarium culmorum and Rhizoctonia solani and the saprotrophic fungus Trichoderma harzianum. Single bacterial strains caused little to moderate growth reduction of fungi (quantified as ergosterol), most probably due to nutrient withdrawal from the media. Growth reduction of fungi by the bacterial mixture was much stronger than that by the single strains. This appeared to be mostly due to competitive interactions between the Pseudomonas and Pedobacter strains. We argue that cohabitation of these strains triggered antibiotic production via interspecific interactions and that the growth reduction of fungi was a side-effect caused by the sensitivity of the fungi to bacterial secondary metabolites. Induction of gliding behavior in the Pedobacter strain by other strains was also observed. Our results indicate that apparently non-antagonistic soil bacteria may be important contributors to soil suppressiveness and fungistasis when in a community context.

Antibiosis↗

Antitumor and antifungal activities in endophytic fungi isolated from pharmaceutical plants Taxus mairei, Cephalataxus fortunei and Torreya grandis.

The purpose of this work was to screen the endophytic fungi having antitumor or antifungal activity, which were isolated from the inner barks of three kinds of pharmaceutical plants, Taxus mairei, Cephalataxus fortunei and Torreya grandis, collected from Fujian province, China. Antitumor activity was studied by the MTT assay and antifungal activity was determined by observing fungal growth inhibition. 13.4% of endophytic fungi fermentation broths displayed cytotoxic activity on HL-60 cells at and below a dilution of 1:50, and 6.4% on KB cells. 52.3% of endophytic fungi fermentation broths displayed growth inhibition on at least one pathogenic fungi, such as Neurospora sp., Trichoderma sp. and Fusarium sp. Among all endophytic fungi isolated, the genus Paecilomyces sp. has the highest positive rate of antitumor and antifungal activity. These results indicate that endophytic fungi could be a promising source for antitumor and antifungal bioactive agents.

Antifungal Agents↗

Ubiquity of lignin-degrading peroxidases among various wood-degrading fungi.

Phanerochaete chrysosporium is rapidly becoming a model system for the study of lignin biodegradation. Numerous studies on the physiology, biochemistry, chemistry, and genetics of this system have been performed. However, P. chrysosporium is not the only fungus to have a lignin-degrading enzyme system. Many other ligninolytic species of fungi, as well as other distantly related organisms which are known to produce lignin peroxidases, are described in this paper. In this study, we demonstrated the presence of the peroxidative enzymes in nine species not previously investigated. The fungi studied produced significant manganese peroxidase activity when they were grown on an oak sawdust substrate supplemented with wheat bran, millet, and sucrose. Many of the fungi also exhibited laccase and/or glyoxal oxidase activity. Inhibitors present in the medium prevented measurement of lignin peroxidase activity. However, Western blots (immunoblots) revealed that several of the fungi produced lignin peroxidase proteins. We concluded from this work that lignin-degrading peroxidases are present in nearly all ligninolytic fungi, but may be expressed differentially in different species. Substantial variability exists in the levels and types of ligninolytic enzymes produced by different white not fungi.

Alcohol Oxidoreductases↗

No significant contribution of arbuscular mycorrhizal fungi to transfer of radiocesium from soil to plants.

The diffuse pollution by fission and activation products following nuclear accidents and weapons testing is of major public concern. Among the nuclides that pose a serious risk if they enter the human food chain are the cesium isotopes 137Cs and 134Cs (with half-lives of 30 and 2 years, respectively). The biogeochemical cycling of these isotopes in forest ecosystems is strongly affected by their preferential absorption in a range of ectomycorrhiza-forming basidiomycetes. An even more widely distributed group of symbiotic fungi are the arbuscular mycorrhizal fungi, which colonize most herbaceous plants, including many agricultural crops. These fungi are known to be more efficient than ectomycorrhizas in transporting mineral elements from soil to plants. Their role in the biogeochemical cycling of Cs is poorly known, in spite of the consequences that fungal Cs transport may have for transfer of Cs into the human food chain. This report presents the first data on transport of Cs by these fungi by use of radiotracers and compartmented growth systems where uptake by roots and mycorrhizal hyphae is distinguished. Independent experiments in three laboratories that used different combinations of fungi and host plants all demonstrated that these fungi do not contribute significantly to plant uptake of Cs. The implications of these findings for the bioavailability of radiocesium in different terrestrial ecosystems are discussed.

Biological Transport↗

Introns and splicing elements of five diverse fungi.

Genomic sequences and expressed sequence tag data for a diverse group of fungi (Saccharomyces cerevisiae, Schizosaccharomyces pombe, Aspergillus nidulans, Neurospora crassa, and Cryptococcus neoformans) provided the opportunity to accurately characterize conserved intronic elements. An examination of large intron data sets revealed that fungal introns in general are short, that 98% or more of them belong to the canonical splice site (ss) class (5'GU...AG3'), and that they have polypyrimidine tracts predominantly in the region between the 5' ss and the branch point. Information content is high in the 5' ss, branch site, and 3' ss regions of the introns but low in the exon regions adjacent to the introns in the fungi examined. The two yeasts have broader intron length ranges and correspondingly higher intron information content than the other fungi. Generally, as intron length increases in the fungi, so does intron information content. Homologs of U2AF spliceosomal proteins were found in all species except for S. cerevisiae, suggesting a nonconventional role for U2AF in the absence of canonical polypyrimidine tracts in the majority of introns. Our observations imply that splicing in fungi may be different from that in vertebrates and may require additional proteins that interact with polypyrimidine tracts upstream of the branch point. Theoretical protein homologs for Nam8p and TIA-1, two proteins that require U-rich regions upstream of the branch point to function, were found. There appear to be sufficient differences between S. cerevisiae and S. pombe introns and the introns of two filamentous members of the Ascomycota and one member of the Basidiomycota to warrant the development of new model organisms for studying the splicing mechanisms of fungi.

Aspergillus nidulans↗

Pneumocystis carinii and specific fungi have a common epitope, identified by a monoclonal antibody.

Because Pneumocystis carinii may be related to fungi, we evaluated the reactivities of monoclonal antibodies raised against P. carinii with a variety of fungi. Fifty-two fungi and six protozoa were evaluated by immunofluorescence. One of three monoclonal antibodies (MAbs) tested (MAb 7D7) reacted with 15 fungi but no protozoa. Saccharomyces cerevisiae showed the strongest reactivity by immunofluorescence. The reactive antigen was characterized for four fungi by the immunoblot technique. In all cases the antigen that was reactive with MAb 7D7 was larger than the P. carinii antigens that reacted with 7D7. In further studies with P. carinii, Aspergillus species, and S. cerevisiae, we found that MAb 7D7 reacted with a carbohydrate component in all organisms. The presence of an epitope that is common to P. carinii and a number of fungi further supports the fungal nature of P. carinii.

Antibodies, Monoclonal↗

Transformation in fungi.

Transformation with exogenous deoxyribonucleic acid (DNA) now appears to be possible with all fungal species, or at least all that can be grown in culture. This field of research is at present dominated by Saccharomyces cerevisiae and two filamentous members of the class Ascomycetes, Aspergillus nidulans and Neurospora crassa, with substantial contributions also from fission yeast (Schizosaccharomyces pombe) and another filamentous member of the class Ascomycetes, Podospora anserina. However, transformation has been demonstrated, and will no doubt be extensively used, in representatives of most of the main fungal classes, including Phycomycetes, Basidiomycetes (the order Agaricales and Ustilago species), and a number of the Fungi Imperfecti. The list includes a number of plant pathogens, and transformation is likely to become important in the analysis of the molecular basis of pathogenicity. Transformation may be maintained either by using an autonomously replicating plasmid as a vehicle for the transforming DNA or through integration of the DNA into the chromosomes. In S. cerevisiae and other yeasts, a variety of autonomously replicating plasmids have been used successfully, some of them designed for use as shuttle vectors for Escherichia coli as well as for yeast transformation. Suitable plasmids are not yet available for use in filamentous fungi, in which stable transformation is dependent on chromosomal integration. In Saccharomyces cerevisiae, integration of transforming DNA is virtually always by homology; in filamentous fungi, in contrast, it occurs just as frequently at nonhomologous (ectopic) chromosomal sites. The main importance of transformation in fungi at present is in connection with gene cloning and the analysis of gene function. The most advanced work is being done with S. cerevisiae, in which the virtual restriction of stable DNA integration to homologous chromosome loci enables gene disruption and gene replacement to be carried out with greater precision and efficiency than is possible in other species that show a high proportion of DNA integration events at nonhomologous (ectopic) sites. With a little more trouble, however, the methodology pioneered for S. cerevisiae can be applied to other fungi too. Transformation of fungi with DNA constructs designed for high gene expression and efficient secretion of gene products appears to have great commercial potential.

DNA, Fungal↗

Keratinopathogenic mould fungi and dermatophytes in healthy and diseased hooves of horses.

Specimens of hoof horn from 187 horses were examined for a possible relationship between clinically affected hooves and the occurrence of pathogenic fungi. Specimens were taken from the coronary band and from the stratum externum and medium of the coronary horn and transferred on to Sabouraud dextrose agar, with and without cycloheximide, and incubated at 28 degrees C. Dermatophytes and mould fungi were identified by their macroscopic and microscopic characteristics. The 732 isolates could be assigned to 26 species of moulds, two different species of the dermatophyte Microsporum and three different species of the dermatophyte Trichophyton. Depending on their pathogenic potential they were assigned to three groups: (i) fungi known to be keratinopathogenic (Acremonium blochii, Alternaria alternata, Alternaria chlamydospora, Geotrichum candidum, Microsporum ferrugineum, Microsporum gypseum, Scopulariopsis brevicaulis, Trichophyton species, Trichophyton mentagrophytes, Trichophyton schönleinii, 57 isolates), (ii) a group of uncertain pathogenicity (223 isolates), and (iii) a group of non-pathogenic species (452 isolates). Eighty per cent of the samples from horses with hoof horn lesions and 66.7 per cent of the samples from horses with slightly affected hoof horn contained fungi of the keratinopathogenic group, whereas only 8.9 per cent of the samples from horses with healthy hoof horn contained fungi of this group. There were no significant correlations between the clinical data and the age, sex or breed of the horses or their bedding and hygiene. Twelve species of fungi were isolated from the air in the horses' stables, but none of them belonged to the keratinopathogenic group.

Animals↗

Calcium-binding proteins in fungi and higher plants.

Calcium has long been known to be required for many vital processes in fungi and plants. High levels of calcium are found in cell walls, vacuoles, and most organelles. In contrast, very low levels of calcium are present in the cytosol of fungal and plant cells. The most recent evidence indicates that calcium is a true second messenger in fungi and plants. Because cyclic AMP does not appear to be a second messenger in plants, calcium is the only known second messenger. Calcium-binding proteins are involved in the events that accompany the action of calcium as a second messenger; three types have been identified in fungi and plants. The first group includes several proteins that bind 45 Ca2+ and are not known to have any enzymatic activity. A second type includes the many enzymes from fungi and plants stimulated by millimolar levels of calcium. The third type of calcium-binding protein, calmodulin, responds to micromolar levels of Ca2+ by binding to certain enzymes and stimulating them. Calmodulin has been detected in every eukaryote thus far examined. The amino acid composition of several fungal and plant calmodulins have been elucidated and found to be very similar to calmodulin from animals. Eight enzymes from fungi and plants have been reported to be regulated either directly or indirectly by calmodulin. Calmodulin antagonists have been used to study the possible involvement of calmodulin in many cellular processes in fungi and plants.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium-Binding Proteins↗

General assessment of the influence of a municipal landfill site and environmental factors on the occurrence of keratinolytic fungi in soil.

The study was to generally determine the influence of a municipal landfill site and environmental factors on the distribution of keratinolytic fungi in soil. The landfill site in Sosnowiec was selected for examination. Keratinolytic fungi occurred abundantly in soils of the landfill site examined and its surrounding area. Of 495 soil samples (Petri dishes) examined, 379 (76.56%) were found to be positive for keratinolytic fungi. Altogether, 1131 strains from 26 species were isolated from the samples. Among the fungi, some species with pathogenic properties (Microsporum racemosum, M. cookei, M. gypseum, Aphanoascus fulvescens and Scopulariopsis brevicaulis) were recorded. The influence of environmental factors on the qualitative and quantitative composition of keratinolytic fungi in the soils was complex. Among these factors, exchangeable acidity (pH in 1 M KCl, in particular), faecal bacterial contamination and the level of water deficit in soil were the most important. The conclusion has been drawn that municipal landfill sites are the sources of potentially pathogenic fungi with keratinolytic properties.

Environment↗

Ergosterol biosynthesis in novel melanized fungi from hypersaline environments.

Halotolerant and halophilic melanized fungi were recently described in hypersaline waters. A close study of the sterol composition of such fungi, namely Hortaea werneckii, Alternaria alternata, Cladosporium sphaerospermum, Cladosporium sp., and Aureobasidium pullulans revealed the dominance of ergosterol and the presence of 29 intermediates of its biosynthesis pathway. The presence or absence of intermediates from distinct synthesis routes gave insight into the operative synthetic pathways from 4,4,14-trimethylcholesta-8,24-dien-3 beta-ol (lanosterol) to ergosterol in melanized fungi and in Saccharomyces cerevisiae, a reference yeast cultured in parallel. In all studied melanized fungi, initial methylation at C-24 took place before C-14 and C-4 demethylation, involving a different reaction sequence from that observed in S. cerevisiae. Further transformation was observed to occur through various routes. In A. alternata, isomerization at C-7 takes place prior to desaturation at C-5 and C-22, and methylene reduction at C-24. In addition to these pathways in Cladosporium spp., H. werneckii, and A. pullulans, ergosterol may also be synthesized through reduction of the C-24 methylene group before desaturation at C-5 and C-22 or vice versa. Moreover, in all studied melanized fungi except A. alternata, ergosterol biosynthesis may also proceed through C-24 methylene reduction prior to C-4 demethylation. -- Méjanelle, L., J. F. Lòpez, N. Gunde-Cimerman, and J. O. Grimalt. Ergosterol biosynthesis in novel melanized fungi from hypersaline environments. J. Lipid Res. 2001. 42: 352--358.

Alternaria↗

[Marine fungi in sediments of the Sea of Japan (Russian territory) and their biologically active metabolites].

The most abundant marine fungi encountered in various regions of the Sea of Japan belong to the genera Penicillium, Aspergillus, Wardomyces, Trichoderma, Chrysosporium, and Chaetomium. Facultative marine fungi of the genera Scytalidium, Verticillium, and Oidiodendron and obligate marine fungi of the genus Dendryphiella are much less abundant. The composition of marine sediments and the anthropogenic load on them were found to influence the abundance and species diversity of fungi, as well as the occurrence of fungal strains producing hemolytically active substances. The biodiversity of mycobiota and the abundance of hemotoxin-producing fungi in marine sediments may be used to evaluate the anthropogenic load on marine biocenoses. Hemolytic compounds were produced by 57% of the fungi isolated from marine sediments. The hemolytic activity of Chaetomium spiculipilium was revealed in the fraction of the culture liquid containing extracellular fatty acids and pigments. The fatty acid composition of this marine fungus was determined.

Fungi↗

[On the problem og higher fungi origin: Florideae hypothesis].

The history and current state of the hypothesis of the origin of higher fungi (Ascomycetes and Basidiomycetes) and red algae from the common ancestor (Florideae) were analysed. Initially the hypothesis was based on similarity of their vegetative and generative structures (Sachs, 1874; Dodge, 1914; Chadefaund, 1953, 1972, etc.), but later it was confirmed by ultrastructural data (Demoulin, 1974; Kohlmeyer, 1975). It appears to be very useful for the study of the development of terrestrial flora (Church, 1921; Kohlemeyer, Kohlmeyer, 1979; Atsatt, 1988) and regularities in morphological evolution of higher fungi (Corner, 1964, 1970; Chadefaud, 1960, 1982, 1984). Description of the order Spathulosporales (Kohlmeyer, 1973), combining the characters of Ascomycetes and parasitic Florideae, was one of the most important fact leading to the wide recognition of the hypothesis in 1970-80s (Cavalier-Smith, 1978; Taylor, 1978; Dodge, 1980; Hawksworth, 1982; Goff, 1983; Coff, Coleman, 1985). Today, however, Florideae hypothesis is not confirmed by molecular data and replaced by alternative hypothesis of Eumycota origin. Summarizing data on molecular systematic of fungi, one could affirm with confidence: 1). Chitincontaining fungi are closer to multicellular animals and green plants than to Rhodophyta; 2). Rhodophyta and Chlorophyta are monophyletic group; 3). There is no single-valued molecular data on taxonomic distance between higher fungi, Chytridiomycetes; Rhodophyta, Chlorophyta and Metazoa. Thus, the current data could not testify against Florideae hypothesis. It is possible to adjust them with the idea of B.M. Kozo-Polyansky (1927) about existence of "Chloroflorodeae" group that is original for terrestrial flora; the hypothesis about closeness of Chlorophyta and Rhodophyta, as well as Chlorophyta and Eumycota, does not contradict molecular data (Stiller, Hall, 1997). The author believes that we need molecular study of the whole "stem" of chlorobionta, especially groups that are close to its basis. It will will help to understand the relationships between Rhodophyta, giant conglomeration of "green algae", chitin-containing fungi, Prasinophyceae and different groups of zoomastigot with maximal resolution--the level of orders or groups of orders. Only such investigation could provide the material for molecular support of phylogenetic constructions. Up to moment Florideae hypothesis is able to exist together with the other ideas.

Ascomycota↗

[Establishment of symbiotic system for Anoectochilus roxburghii (Wall.) Lindl. and endophytic fungi].

OBJECTIVE: Setting up a symbiotic system for Anoectochilus roxburghii and endophytic fungi, so as to study the relationship between them and try to set up new cultural methods for A. roxburghii. METHOD: A. roxburghii and its endophytic fungi were cultured together on five kinds of media in flasks. The growth of the plantlets and fungi were observed. Symbiotic conditions were selected according to symbiotic characteristics. RESULTS: A symbiotic system for A. roxburghii and endophytic fungi was set up. The optimum composition of the system induded NH4NO3 825 mg.L-1, KNO3 950 mg.L-1, MgSO4 185 mg.L-1, and inositol 100 mg.L-1, other organic components being 2/3 times those of MS medium, sugrose 15 g.L-1, and other components being the same as those of MS medium, agar 9 g.L-1, pH 5.8. The culture was effected at 24-25 degrees C under cool white fluorescent light (150 lx) for a photoperiod of 11 hours. CONCLUSION: A. roxburghii and endophytic fungi can grow well together and form endomycorrhiza in the symbiotic system. Moreover, endophytic fungi help stimulate the growth and development of A. roxburghii.

Culture Media↗

[Effect of environmental factors on the growth and fatty acid composition of five endophytic fungi from Sapium sebiferum].

Five fungi species including Rhizoctonia sp., Sclerotium sp., Acremoniell sp., Chaetomium sp. and Pestalotiopsis sp. screened from the inner bark of Sapium sebiferum (Lével) Roxb. were used to study their growth under different environmental factors, with the biomass and fatty acid composition as main indexes to elucidate the relationship between plant and its endophytic fungi. Compared with that from synthesized medium, a higher biomass was harvested when the fungi were cultured on liquid potato dextrose medium; the unsaturation index of fatty acid (IUFA) was lower, and the main fatty acids in the fungi were linoleic acid, palmitic acid and oleic acid. Compared with the fungi grown on the synthesized medium without Sapium sebiferum's extract, the Sclerotium sp. growth was promoted, and the others were inhibited when the extract was added to the synthesized medium. At the same time, the IUFA were increased. When 0-1.0 mol.L-1 NaCl was added in Sclerotium sp.'s culture medium, the biomass was not changed notably; IUFA in 0-0.5 mol.L-1 NaCl was not significantly different, while in the range of 0.6-1.0 mol.L-1 NaCl, the IUFA was decreased with increasing concentration of NaCl. It indicated that fungus had a stronger tolerance to salt. Sclerotium sp.'s growth was promoted, and the IUFA was increased when plant oil was added in the synthesized medium. The biomass was the highest by adding 1.5% plant oil. The results mentioned above were some way related to the symbiotic relations between the endophytic fungi and their host plant.

Biomass↗