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Genetic processes in arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal (AM) fungi (Glomeromycota) colonize roots of the majority of land plants and facilitate their mineral nutrient uptake. Consequently, AM fungi play an important role in terrestrial ecosystems and are becoming a component of sustainable land management practices. The absence of sexual reproductive structures in modern Glomeromycota combined with their long evolutionary history suggest that these fungi may represent an ancient asexual lineage of great potential interest to evolutionary biology. However, many aspects of basic AM fungal biology, including genome structure, within-individual genetic variation, and reproductive mode are poorly understood. These knowledge gaps hinder research on the mechanisms of AM fungal interactions with individual plants and plant communities, and utilization of AM fungi in agricultural practices. I present here the current state of research on the reproduction in AM fungi and indicate what new findings can be expected in the future.

Fungi↗

Sampling for indoor fungi.

BACKGROUND: A great deal of concern has arisen recently regarding the potential adverse effects of indoor fungi. Our understanding of this complex problem has been hampered by a lack of standardized protocols for performing an indoor assessment for fungi. Without such standards, it is difficult to compare results from one study with those from another or to measure the effect of indoor fungal contamination on a building and its occupants. METHODS: We reviewed the medical literature and here describe a hypothesis-driven approach to planning, sampling, and interpreting the results of indoor assessments for fungi. RESULTS: Fungi cause 3 primary adverse effects: (1) they can damage a building, (2) they can render a building unpleasant to live in by looking and smelling bad, and (3) they might cause adverse health effects in sensitive individuals. Sampling methods used to test hypotheses include air sampling for spores, measurement of allergens in house dust, and determination of microbially generated volatile organic compounds, ergosterols, glucans, and mycotoxins, as well as environmental conditions that lead to fungal contamination. CONCLUSIONS: Standardized approaches for performing and reporting assessments of indoor fungi are essential if our understanding of this complex field is to improve.

Air Microbiology↗

Chronic rhinosinusitis: an enhanced immune response to ubiquitous airborne fungi.

BACKGROUND: Chronic rhinosinusitis (CRS) is one of the most common long-term illnesses in the United States. The etiology of CRS is unknown, and no effective treatment has been established. OBJECTIVE: We investigated the hypothesis that abnormal immunologic responses to ubiquitous airborne fungi contribute to the pathogenesis of this disease. METHODS: The proliferative and cytokine responses of PBMCs to extracts from 4 common airborne fungi-including Alternaria , Aspergillus , Cladosporium , and Penicillium -were examined by in vitro culture. Serum specimens were tested for specific IgE and IgG to these fungi. RESULTS: PBMCs from approximately 90% of the patients with CRS, but not those from normal individuals, produced both IL-5 and IL-13 when exposed to Alternaria. Furthermore, PBMCs from patients with CRS produced significantly more IFN-gamma than PBMCs from normal individuals in response to Alternaria (median, 553 pg/mL vs 98 pg/mL; P < .01). Levels of serum IgG antibodies to Alternaria and Cladosporium were clearly increased in patients with CRS compared with normal individuals ( P < .01). Less than 30% of the patients with CRS had specific IgE antibodies to Alternaria or Cladosporium. The increased humoral (serum IgG) response strongly correlated with the increased cellular (IL-5 production) response to Alternaria ( r = 0.619; P < .01). CONCLUSION: Patients with CRS show exaggerated humoral and cellular responses, both T(H)1 and T(H)2 types, to common airborne fungi, particularly Alternaria. The anomalous immune and inflammatory responses to ubiquitous fungi may explain the chronicity of airway inflammation in CRS.

Adult↗

An improved method for the selective detection of fungi in hospital waters by solid phase cytometry.

Yeast cells and mould spores can be fluorescently labelled with the viability stain carboxyfluorescein diacetate (CFDA) and detected on a membrane filter by laser scanning (solid phase cytometry, SPC). Although the selectivity of an existing commercial SPC procedure for fungi is ensured by using a 2 microm pore size membrane filter and a pre-incubation on a proprietary spore swelling/activation medium, some bacteria are still co-detected. In the present study, the selectivity for fungi has been enhanced by combining the green fluorescent CFDA with a second red fluorescent label, i.e. TRITC-concanavalin A, targetting fungal but not commonly bacterial cells. Additional improvements resulted from the prolongation of the pre-incubation and from the extra-rinsing of the membrane filter. The improved method was applied to detect fungi in hospital waters, dialysis fluids and endoscopic rinse waters. In general, SPC detected more fungi in water than plate methods. The occurrence of fungi in dialysis fluid and endoscopic rinse water was rare. Evidence for the presence of fungal viable but non-culturable (VBNC) cells in water was weak.

Colony Count, Microbial↗

Chitinolytic activities of ericoid mycorrhizal and other root-associated fungi from Epacris pulchella (Ericaceae).

Ericoid mycorrhizal endophytes and other root-associated fungi from Epacris pulchella (Ericaceae) in an eastern Australian sclerophyll forest, along with Hymenoscyphus ericae, were tested for their abilities to produce extracellular chitinolytic activities during growth in axenic culture. Two root-associated fungi produced activities that were active against only a monomeric 4-methylumbelliferyl (4-MU) glycoside of N-acetylglucosamine, suggesting exo-acting beta-N-acetylhexosaminidase (EC 3.2.1.52) activity. All ericoid mycorrhizal fungi and two root-associated fungi produced activities against dimeric and trimeric 4-MU glycosides of N-acetylglucosamine, suggesting production of chitobiosidase and endo-acting chitinase (EC 3.2.1.14) respectively in addition to beta-N-acetylhexosaminidase. Specific activities for all ericoid mycorrhizal fungi, including H. ericae, were of the same order of magnitude, suggesting that their chitinolytic potential is broadly similar. Chitinase activities were only produced by an ericoid mycorrhizal fungus when chitin was included in the medium, however, no activity was produced if glucose was also present in the medium.

Chitinases↗

Culturable airborne fungi in outdoor environments in Beijing, China.

Airborne fungi are being proposed as a cause of adverse health effects. They may adversely affect human health through allergy, infection, and toxicity. Moreover, they have a great influence on urban air quality in Beijing. In this study, a systematical survey on the culturable airborne fungi was carried out for 1 year in Beijing urban area. Fungal samples were collected for 3 min, three times each day, and continued for three consecutive days of each month with FA-1 sampler from three sampling sites. Results showed that the culturable fungal concentrations ranged from 24 CFU (Colony forming units) /m3 to 13960 CFU/m3, and the mean and median was 1165 CFU/m3 and 710 CFU/m3, respectively. Fungal concentrations in the greener area around the Research Center for Eco-Environmental Sciences (RCEES) and Beijing Botanical Garden (BBG) were significantly higher than in the densely urban and highly trafficked area of Xizhimen (XZM) (***P<0.001), but no significant difference was found between RCEES and BBG (P>0.05). The variation of fungal concentrations in different seasons was significant in RCEES and BBG, where the concentrations were higher in Summer and Autumn, and lower in Spring and Winter. However, there were no significant differences in fungal concentrations between the Spring and the Winter for three sampling sites (P>0.05). Fourteen genera, including 40 species of culturable fungi, were identified in this study. Penicillium, with the most abundant species, which comprised more than 50% of the total isolated fungal species. Cladosporium were the most dominant fungal group, and contributed to more than one third of the total fungal concentration, followed by non-sporing isolates, Alternaria, Pencillium and Asperigillus. The concentration percentage of Cladosporium was significantly higher in RCEES than in XZM (*P<0.05), and the concentration percentages of Penicillium (**P<0.01) and Aspergillus (*P<0.05) were higher in XZM than in RCEES and in BBG. For other groups' concentration percentages, no significant differences were observed among the sampling sites. The distribution pattern of airborne fungi presented log-normal distribution. The highest proportion of culturable fungi was detected in stage 4 (2.0-3.5 microm), and the lowest was in stage 6 (<1.0 microm).

Air Microbiology↗

Iron uptake by fungi: contrasted mechanisms with internal or external reduction.

Almost all iron uptake by fungi involves reduction from Fe(III) to Fe(II) in order to facilitate ligand exchange. This leads to two mechanisms: uptake before reduction, or reduction before uptake. Many fungi secrete specific hydroxamate siderophores when short of iron. The mechanism with uptake before reduction is described in the context of siderophore synthesis and usage, since it applies to many (but not all) siderophores. The hydroxamate functional group is synthesized from ornithine by N5 hydroxylation and acylation. In most fungal siderophores, two or three modified ornithines are joined together by a non-ribosomal peptide synthetase. The transcription of these genes is regulated by an iron activated repressor. There is evidence that the iron-free siderophore may be stored in intracellular vesicles until secretion is required. After loading with iron, re-entry is likely to be via a proton symport. In some fungi, siderophores are used for iron storage. The iron is liberated by an NADPH-linked reductase. The second mechanism starts with Fe(III) reduction. In yeast, this is catalysed by an NADPH-linked transmembrane reductase, which has homology with the NADPH oxidase of neutrophils. There are two closely similar reductases with overlapping roles in Fe(III) and Cu(II) reduction, while the substrates for reduction include Fe(III)-siderophores. External reductants, which may be important in certain fungi, include 3-hydroxyanthranilic acid, melanin, cellobiose dehydrogenase and 2,5-dimethylhydroquinone. In yeast, a high-affinity iron uptake pathway involves reoxidation of Fe(II) to Fe(III), probably to confer specificity for iron. This is catalysed by a copper protein which has homology with ceruloplasmin, and is closely coupled to Fe(III) transport. The transcription of these genes is regulated by an iron-inhibited activator. Because of its copper requirement, the high-affinity pathway is blocked by disruption of genes for copper metabolism. A low-affinity uptake transports Fe(II) directly and is important in anoxic growth. In many fungi, mechanisms with internal or external reduction are both important. The external reduction is applicable to almost any Fe(III) complex, while internal reduction is more efficient at low iron but requires a siderophore permease through which toxins might enter. Both mechanisms require close coupling of Fe(III) reduction and Fe(II) utilization in order to minimize production of active oxygen.

Biological Transport↗

Nematophagous fungi in fresh feces of cattle in the Mata region of Minas Gerais state, Brazil.

The capacity to survive gut passage is one of the desirable characteristics for nematophagous fungi to be considered potential biological control agents of gastrointestinal nematodes of livestock. From April 1995 to April 1996, a pool of 10 cow fecal samples and 10 individual samples of feces from heifers, which were raised under partial (cows) or total (heifers) confinement in the Mata Region of Minas Gerais State, Brazil, were examined monthly for the presence of nematophagous fungi. A total of 10 isolates was found in the survey. Eight isolates were recovered from the pooled samples of cow feces and two from the individual samples of heifers. Fungi were present in the cow feces during the dry months of August (two isolates of Arthrobotrys oligospora and one Monacrosporium eudermatum) and September (one isolate of Harposporium lilliputanum and one of M. gephyropagum). Fungi were also recovered at the beginning and middle of the rainy season: one isolate of A. musiformis in October, and one isolate of M. gampsosporum and one unidentified fungus which produced septate hyphae and adhesive buds in December. In the individual samples collected from heifers, fungi were present only in the months of September (end of dry season) and March (end of rainy season). One isolate each of H. lilliputanum and A. oligospora were found, respectively. Additional studies to further characterize these isolates should be encouraged.

Animals↗

Iron gathering by zoopathogenic fungi.

Iron is a metal required by most microorganisms and is prominently used in the transfer of electrons during metabolism. The gathering of iron is, then, an essential process and its fulfillment becomes a crucial pathogenetic event for zoopathogenic fungi. Iron is rather unavailable because it occurs on the earth's surface in its insoluble ferric form in oxides and hydroxides. In the infected host iron is bound to proteins such as transferrin and ferritin. Solubilization of ferric iron is the major problem confronting microorganisms. This process is achieved by two major mechanisms: ferric reduction and siderophore utilization. Ferric reductase is frequently accompanied by a copper oxidase transport system. There is one example of direct ferric iron transport apparently without prior reduction. Ferric reduction may also be accomplished by low molecular mass compounds. Some fungi have evolved a process of iron acquisition involving the synthesis of iron-gathering compounds called siderophores. Even those fungi that do not synthesize siderophores have developed permeases for transport of such compounds formed by other organisms. Fungi can also reductively release iron from siderophores and transport the ferrous iron often by the copper oxidase transport system. There is a great diversity of iron-gathering mechanisms expressed by pathogenic fungi and such diversity may be found even in a single species.

Ferritins↗

Clinical use of immunoassays in assessing exposure to fungi and potential health effects related to fungal exposure.

OBJECTIVE: To review and summarize current evidence regarding the proper role of immunoassays in clinical assessments of exposure to fungi and health effects related to fungal exposure. DATA SOURCES: We reviewed relevant scientific investigations and previously published reviews concerning this topic. STUDY SELECTION: The authors' clinical, laboratory, and public health experiences were used to evaluate relevant data for scientific merit. RESULTS: Testing to determine the presence of IgE to specific fungi may be a useful component of a complete clinical evaluation in the diagnosis of illnesses that can be caused by immediate hypersensitivity such as allergic rhinitis and asthma. Detection of IgG to specific fungi has been used as a marker of exposure to agents that may cause illnesses such as hypersensitivity pneumonitis. However, the ubiquitous nature of many fungi and the lack of specificity of fungal antigens limit the usefulness of these types of tests in the evaluation of potential building-related illness and fungal exposure. Specific serologic tests (such as tests for cryptococcal antigen, coccidioidal antibody, and Histoplasma antigen) have been shown to be useful in the diagnosis of some fungal infections, but these are the exception not the rule. CONCLUSIONS: There is currently not enough scientific evidence to support the routine clinical use of immunoassays as a primary means of assessing environmental fungal exposure or health effects related to fungal exposure. Health care providers who care for persons expressing concerns about the relationship of symptoms to potential exposure to fungi are advised to use immunoassay results with care and only as an adjunct to a comprehensive approach to patient care.

Animals↗

Ecology of cycloheximide-resistant fungi in field soils receiving raw city wastewater or normal irrigation water.

The effect of raw city wastewater irrigation on biodiversity and population densities of a cycloheximide-resistant (CH) fungal community was studied in 13 field soils receiving either raw city wastewater or normal irrigation, and in raw city wastewater in the Nablus area, using the hair baiting technique (HBT) and a surface soil dilution plating (SSDP) technique. Three of these fields [one had been receiving raw city wastewater for more than ten years and was designated a heavily polluted field, and the other 2 were cultivated for the first time and were either irrigated with raw city wastewater (newly polluted field) or normal irrigation water (nonpolluted)], were sampled 4-7 times over a 9-month period. The other ten fields, which had been under raw city wastewater irrigation for more than 10 years, were sampled only once. Fifty-seven CH-resistant species belonging to 18 genera were recovered, of which 49 species were recovered from soil habitats and 28 species from raw city wastewater. The HBT had shown to be more efficient in the isolation of pathogenic and potentially pathogenic fungi including dermatophytes. A higher percentage of this group of fungi was recovered from the three main field soils studied using HBT (70% of all isolates), than the SSDP (35.5%); no dermatophytes were recovered by the SSDP method. Two dermatophytes (Microsporum gypseum, and Trichophyton ajelloi), and five more fungi (Arthroderma cuniculi, A. curreyi, Chrysosporium keratinophilum, C. tropicum, C. pannorum), were recovered from these habitats. Wastewater irrigation seemed to have affected the fungal population densities, with the highest population densities being found in the heavily polluted field soil, while lower population densities were found in the nonpolluted field soil. Increases in organic matter were also observed as a result of sewage effluent irrigation. However, basic similarities in the biodiversity of CH-resistant fungal communities existed in nonpolluted and polluted field soils, and raw city wastewater. Comparable numbers of fungal species were recovered from the three main field soils. The species most commonly found in those habitats included: Alternaria alternata, Aspergillus candidus, Geotrichum candidum, and Paecilomyces lilacinus. Field soils receiving either raw city wastewater or normal irrigation water, were found to be rich in pathogenic and potentially pathogenic CH-resistant fungi, including dermatophytes, with raw city wastewater yielding the highest percentage (81%), followed by the newly wastewater irrigated field (77.7%), the nonpolluted field (67%), and the heavily polluted field (63.4%) Hygienic measures should therefore be taken to control the spread of these fungi in the environment of human communities, and to avoid mycotic infections among farmers.

Agriculture↗

A review of soybean (Glycine max) seed, pod, and flower mycofloras in North America, with methods and a key for identification of selected fungi.

A review of the fungi associated with soybean seeds, pods, or flowers was conducted in North America. Species of Deuteromycetes are the most common fungi in each of the soybean flower organs followed by the Ascomycetes and Phycomycetes which comprise about one-fourth of the total mycoflora. Eighty genera and about 135 or more species occur in seeds, pods, or flowers. With regard to numbers of taxa from separate mycofloras, 63 genera and about 108 or more species occur in seeds, 65 genera and about 88 or more species occur in pods, and 36 genera and approximately 47 or more species occur in flowers. Most of the fungi which occur in flowers can be cultured from pods, and the majority of those fungi occur in seeds. Methods for and a key are provided for the identification of the 30 most important selected fungi.

Ascomycota↗

Transformation of 2,4,6-trichlorophenol by the white rot fungi Panus tigrinus and Coriolus versicolor.

The toxicity of thirteen isomers of mono-, di-, tri- and pentachlorophenols was tested in potato-dextrose agar cultures of the white rot fungi Panus tigrinus and Coriolus versicolor. 2,4,6-Trichlorophenol (2,4,6-TCP) was chosen for further study of its toxicity and transformation in liquid cultures of these fungi. Two schemes of 2,4,6-TCP addition were tested to minimize its toxic effect to fungal cultures: stepwise addition from the moment of inoculation and single addition after five days of growth. In both cases the ligninolytic enzyme systems of both fungi were found to be responsible for 2,4,6-TCP transformation. 2,6-Dichloro-1,4-hydroquinol and 2,6-dichloro-1,4-benzoquinone were found as products of primary oxidation of 2,4,6-TCP by intact fungal cultures and purified ligninolytic enzymes, Mn-peroxidases and laccases of both fungi. However, primary attack of 2,4,6-TCP in P. tigrinus culture was conducted mainly by Mn-peroxidase, while in C. versicolor it was catalyzed predominantly by laccase, suggesting a different mode of regulation of these enzymes in the two fungi.

Benzoquinones↗

Evidence for the evolution of multiple genomes in arbuscular mycorrhizal fungi.

Ancient asexuals directly contradict the evolutionary theories that explain why organisms should evolve a sexual life history. The mutualistic, arbuscular mycorrhizal fungi are thought to have been asexual for approximately 400 million years. In the absence of sex, highly divergent descendants of formerly allelic nucleotide sequences are thought to evolve in a genome. In mycorrhizal fungi, where individual offspring receive hundreds of nuclei from the parent, it has been hypothesized that a population of genetically different nuclei should evolve within one individual. Here we use DNA-DNA fluorescent in situ hybridization to show that genetically different nuclei co-exist in individual arbuscular mycorrhizal fungi. We also show that the population genetics techniques used in other organisms are unsuitable for detecting recombination because the assumptions and underlying processes do not fit the fungal genomic structure shown here. Instead we used a phylogenetic approach to show that the within-individual genetic variation that occurs in arbuscular mycorrhizal fungi probably evolved through accumulation of mutations in an essentially clonal genome, with some infrequent recombination events. We conclude that mycorrhizal fungi have evolved to be multi-genomic.

Cell Nucleus↗

Reconstructing the early evolution of Fungi using a six-gene phylogeny.

The ancestors of fungi are believed to be simple aquatic forms with flagellated spores, similar to members of the extant phylum Chytridiomycota (chytrids). Current classifications assume that chytrids form an early-diverging clade within the kingdom Fungi and imply a single loss of the spore flagellum, leading to the diversification of terrestrial fungi. Here we develop phylogenetic hypotheses for Fungi using data from six gene regions and nearly 200 species. Our results indicate that there may have been at least four independent losses of the flagellum in the kingdom Fungi. These losses of swimming spores coincided with the evolution of new mechanisms of spore dispersal, such as aerial dispersal in mycelial groups and polar tube eversion in the microsporidia (unicellular forms that lack mitochondria). The enigmatic microsporidia seem to be derived from an endoparasitic chytrid ancestor similar to Rozella allomycis, on the earliest diverging branch of the fungal phylogenetic tree.

Chytridiomycota↗

Ecophysiological characterization of common food-borne fungi in relation to pH and water activity under various atmospheric compositions.

The combined effects of pH, water activity (aw), oxygen (O2) and carbon dioxide (CO2) levels on growth and sporulation of 10 common food-borne fungi were studied. The use of a multivariate statistical method (PLS) for the analysis of data showed that the fungi could be grouped according to their physiological response to changes in the four tested factors. Carbon dioxide, aw and pH were found to be the most significant factors describing differences and similarities among the fungi. Maximal inhibitory effect of elevated levels of CO2 (5-25%) and decreased aw (0.99-0.95) varied among the 10 species from 6 to 77% and from 52 to 100%, respectively. Sporulation of the fungi was sensitive to all tested factors. Furthermore, interaction of CO2 and aw displayed a significant effect on sporulation. It was shown that different fungal species associated with the same ecosystem responded similarly to changes in the tested factors. Thus, fungi which are not phylogenetically related may be physiologically related or show a common strategy of life.

Carbon Dioxide↗

The occurrence of fungi in hospital and community potable waters.

The prevalence of fungi was investigated in 126 potable water samples (84 hospital and 42 community samples), in parallel with the standard pollution indicator micro-organisms. Filamentous fungi were isolated from 104 of 126 (82.5%) samples and yeasts from 14 (11.1%), whereas their mean counts were 36.6 and 4.4, respectively. Fungi were isolated from 95.2% of community and 76.2% of hospital water samples, with the difference being statistically significant (P < 0.05), while yeasts were isolated from 9.5 and 11.9%, respectively. Prevailing genera were Penicillium spp., isolated from 64, Aspergillus spp., from 53, and Candida, from nine of the examined samples. Colony-forming units of yeasts were significantly correlated with those of total and faecal coliforms, whereas the counts of filamentous fungi were significantly correlated with total heterotrophic bacteria counts. These results suggest that tap water is a potential transmission route for fungi both in hospitals and the community in the examined region and may pose a health hazard mainly for the immunocompromised host.

Aspergillus↗

Interspecific hybridization in plant-associated fungi and oomycetes: a review.

Fungi (kingdom Mycota) and oomycetes (kingdom Stramenopila, phylum Oomycota) are crucially important in the nutrient cycles of the world. Their interactions with plants sometimes benefit and sometimes act to the detriment of humans. Many fungi establish ecologically vital mutualisms, such as in mycorrhizal fungi that enhance nutrient acquisition, and endophytes that combat insects and other herbivores. Other fungi and many oomycetes are plant pathogens that devastate natural and agricultural populations of plant species. Studies of fungal and oomycete evolution were extraordinarily difficult until the advent of molecular phylogenetics. Over the past decade, researchers applying these new tools to fungi and oomycetes have made astounding new discoveries, among which is the potential for interspecific hybridization. Consequences of hybridization among pathogens include adaptation to new niches such as new host species, and increased or decreased virulence. Hybrid mutualists may also be better adapted to new hosts and can provide greater or more diverse benefits to host plants.

Adaptation, Biological↗