Showing papers on "Mycelium published in 2005"
TL;DR: Results of stable isotope labelling experiments are reported showing that inorganic nitrogen taken up by the fungus outside the roots is incorporated into amino acids, translocated from the extraradical to the intraradical mycelium as arginine, but transferred to the plant without carbon.
Abstract: The discovery of a previously unknown mechanism of nitrogen transfer from the arbuscular mycorrhizal fungi found on the roots of most land plants, to the host plants suggests that this symbiotic relationship may be a much more important factor in the global nitrogen cycle than was thought. The mechanism involves uptake of inorganic nitrogen by the fungus outside the roots, conversion to amino acids within the fungus, then transfer as ammonium ions from the fungal mycelium into the plant. The first event in host recognition by arbuscular mycorrhizal fungi is thought to be hyphal branching. A strigolactone, 5-deoxy-strigol, isolated from Lotus japonicus has now been identified as an inducer of branching. Strigolactones are root metabolites, previously isolated as seed germination stimulants for root parasitic weeds. This finding highlights the close relationship between plant and fungus, and may provide a new strategy for the control of both beneficial fungal symbionts and destructive parasitic weeds in agriculture and natural ecosystems. Most land plants are symbiotic with arbuscular mycorrhizal fungi (AMF), which take up mineral nutrients from the soil and exchange them with plants for photosynthetically fixed carbon. This exchange is a significant factor in global nutrient cycles1 as well as in the ecology2, evolution3 and physiology4 of plants. Despite its importance as a nutrient, very little is known about how AMF take up nitrogen and transfer it to their host plants5. Here we report the results of stable isotope labelling experiments showing that inorganic nitrogen taken up by the fungus outside the roots is incorporated into amino acids, translocated from the extraradical to the intraradical mycelium as arginine, but transferred to the plant without carbon. Consistent with this mechanism, the genes of primary nitrogen assimilation are preferentially expressed in the extraradical tissues, whereas genes associated with arginine breakdown are more highly expressed in the intraradical mycelium. Strong changes in the expression of these genes in response to nitrogen availability and form also support the operation of this novel metabolic pathway in the arbuscular mycorrhizal symbiosis.
917 citations
TL;DR: The results point research on functions of glomalin and GRSP in a new direction, focusing on the contributions this protein makes to the living mycelium, rather than its role once it is released into the soil.
Abstract: Arbuscular mycorrhizal fungi (AMF) produce a protein, glomalin, quantified operationally in soils as glomalin-related soil protein (GRSP). GRSP concentrations in soil can range as high as several mg g−1 soil, and GRSP is highly positively correlated with aggregate water stability. Given that AMF are obligate biotrophs (i.e. depending on host cells for their C supply), it is difficult to explain why apparently large amounts of glomalin would be produced and secreted actively into the soil, since the carbon could not be directly recaptured by the mycelium (and benefits to the AMF via increased soil structure would be diffuse and indirect). This apparent contradiction could be resolved by learning more about the pathway of delivery of glomalin into soil; namely, does this occur via secretion, or is glomalin tightly bound in the fungal walls and only released after hyphae are being degraded by the soil microbial community? In order to address this question, we grew the AMF Glomus intraradices in in vitro cultures and studied the release of glomalin from the mycelium and the accumulation of glomalin in the culture medium. Numerous protein-solubilizing treatments to release glomalin from the fungal mycelium were unsuccessful (including detergents, acid, base, solvents, and chaotropic agents), and the degree of harshness required to release the compound (autoclaving, enzymatic digestion) is consistent with the hypothesis that glomalin is tightly bound in hyphal and spore walls. Further, about 80% of glomalin (by weight) produced by the fungus was contained in hyphae and spores compared to that released into the culture medium, strongly suggesting that glomalin arrives mainly in soil via release from hyphae, and not primarily through secretion. These results point research on functions of glomalin and GRSP in a new direction, focusing on the contributions this protein makes to the living mycelium, rather than its role once it is released into the soil.
357 citations
TL;DR: Arginine (Arg) was the predominant free AA in the ERM, and almost all Arg molecules became labeled within 3 wk of supplying (15)NH(4) (+) to the fungal compartment, which is the most likely form of N transferred to host cells following its generation from Arg breakdown.
Abstract: Nitrogen (N) is known to be transferred from fungus to plant in the arbuscular mycorrhizal (AM) symbiosis, yet its metabolism, storage and transport are poorly understood. In vitro mycorrhizas of Glomus intra-radices and Ri T-DNA-transformed carrot roots were grown in two-compartment Petri dishes. (15)N- and/or (13)C-labeled substrates were supplied to either the fungal compartment or to separate dishes containing uncolonized roots. The levels and labeling of free amino acids (AAs) in the extra-radical mycelium (ERM) in mycorrhizal roots and in uncolonized roots were measured by gas chromatography/mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). Arginine (Arg) was the predominant free AA in the ERM, and almost all Arg molecules became labeled within 3 wk of supplying (15)NH(4) (+) to the fungal compartment. Labeling in Arg represented > 90% of the total (15)N in the free AAs of the ERM. [Guanido-2-(15)N]Arg taken up by the ERM and transported to the intra-radical mycelium (IRM) gave rise to (15)N-labeled AAs. [U-(13)C]Arg added to the fungal compartment did not produce any (13)C labeling of other AAs in the mycorrhizal root. Arg is the major form of N synthesized and stored in the ERM and transported to the IRM. However, NH(4) (+) is the most likely form of N transferred to host cells following its generation from Arg breakdown.
274 citations
TL;DR: The fungus Beauveria caledonica was highly tolerant to toxic metals and solubilized cadmium, copper, lead, and zinc minerals, converting them into oxalates, suggesting that a ligand-promoted mechanism was the main mechanism of mineral dissolution.
Abstract: The fungus Beauveria caledonica was highly tolerant to toxic metals and solubilized cadmium, copper, lead, and zinc minerals, converting them into oxalates. This fungus was found to overexcrete organic acids with strong metal-chelating properties (oxalic and citric acids), suggesting that a ligand-promoted mechanism was the main mechanism of mineral dissolution. Our data also suggested that oxalic acid was the main mineral-transforming agent. Cadmium, copper, and zinc oxalates were precipitated by the fungus in the local environment and also in association with the mycelium. The presence of toxic metal minerals often led to the formation of mycelial cords, and in the presence of copper-containing minerals, these cords exhibited enhanced excretion of oxalic acid, which resulted in considerable encrustation of the cords by copper oxalate hydrate (moolooite). It was found that B. caledonica hyphae and cords were covered by a thick hydrated mucilaginous sheath which provided a microenvironment for chemical reactions, crystal deposition, and growth. Cryo-scanning electron microscopy revealed that mycogenic metal oxalates overgrew parental fungal hyphae, leaving a labyrinth of fungal tunnels within the newly formed mineral matter. X-ray absorption spectroscopy revealed that oxygen ligands played a major role in metal coordination within the fungal biomass during the accumulation of mobilized toxic metals by B. caledonica mycelium; these ligands were carboxylic groups in copper phosphate-containing medium and phosphate groups in pyromorphite-containing medium.
255 citations
TL;DR: Chitinous material extracted from mycelia of Aspergillus niger and Mucor rouxii grown in yeast peptone dextrose broth and tested for antibacterial and eliciting properties significantly reduced lesions caused by Botrytis cinerea and Penicillium expansum in harvested apples.
Abstract: Chitinous material was extracted from mycelia of Aspergillus niger and Mucor rouxii grown in yeast peptone dextrose broth for 15 and 21 days, respectively. The extracted material was characterized for purity, degree of acetylation, and crystallinity and tested for antibacterial and eliciting properties. The maximum glucosamine level determined in the mycelium of A. niger was 11.10% dw and in the mycelium of M. rouxii was 20.13% dw. On the basis of the stepwise extraction of freeze-dried mycelia, it appeared that M. rouxii mycelia contained both chitin and chitosan, whereas A. niger contained only chitin. The yields of crude chitin from A. niger and M. rouxii were 24.01 and 13.25%, respectively, and the yield of chitosan from M. rouxii was 12.49%. Significant amounts (7.42-39.81%) of glucan were associated with chitinous compounds from both species and could not be eliminated by the extraction method used. The degrees of acetylation were determined to be 76.53 and 50.07% for chitin from A. niger and M. rouxii, respectively, and 19.5% for M. rouxii chitosan. The crystallinity of fungal chitin and chitosan was estimated to be less intense than in corresponding materials from shrimp shells. The extracted chitin and chitosan in a concentration of 0.1% reduced Salmonella Typhimurium DT104 2576 counts by 0.5-1.5 logs during a 4 day incubation in tryptic soy broth at 25 °C. Furthermore, all tested chitinous materials from fungal sources significantly reduced lesions caused by Botrytis cinerea and Penicillium expansum in harvested apples.
233 citations
TL;DR: Findings indicate that the uptake of P by the fungus and its transfer to the host is also stimulated by the transfer of carbon from plant to fungus across the mycorrhizal interface.
Abstract: Summary
• The influence of carbohydrate availability to mycorrhizal roots on uptake, metabolism and translocation of phosphate (P) by the fungus was examined in axenic cultures of transformed carrot (Daucus carota) roots in symbiosis with Glomus intraradices.
• 14C-labelled carbohydrates, 33P-phosphate and energy dispersive X-ray microanalysis were used to follow the uptake and transfer of C and P in the arbuscular mycorrhizal (AM) symbiosis.
• The uptake of P by the extraradical mycelium (ERM) and its translocation to the mycorrhizal roots was stimulated and the metabolic and spatial distribution of P within the fungus were altered in response to increased carbohydrate availability. Sucrose supply resulted in a decrease of polyphosphates and an increased incorporation into phospholipids and other growth-related P pools and also caused elevated cytoplasmic P levels in the intraradical mycelium (IRM) within the root and higher cytoplasmic P levels in the root cortex.
• These findings indicate that the uptake of P by the fungus and its transfer to the host is also stimulated by the transfer of carbon from plant to fungus across the mycorrhizal interface.
177 citations
TL;DR: The overall transcriptome analysis provided information about sequences related to the cell cycle, stress response, drug resistance, and signal transduction pathways of the pathogen.
176 citations
TL;DR: A role of GintZnT1 in Zn compartmentalization and in the protection of G. intraradices against Zn stress is suggested.
171 citations
TL;DR: It is proposed that this actinomycete, strain FXJ46(T), was isolated from cypress forest soil in northern China and shown to have chemotaxonomic and morphological properties consistent with streptomycetes, be classified as a novel species of the genus Streptomyces.
Abstract: An actinomycete, strain FXJ46T, was isolated from cypress forest soil in northern China and shown to have chemotaxonomic and morphological properties consistent with streptomycetes. It developed greyish aerial mycelium and pinkish-brown substrate mycelium on oatmeal agar. Phylogenetic analyses based on an almost complete 16S rRNA gene sequence of the strain and on the 120 nucleotide variable γ-region of this molecule showed that it formed a distinct (but closely associated) line with Streptomyces griseoaurantiacus DSM 40430T in Streptomyces trees. However, the DNA–DNA relatedness between the two strains was only 48·8 %. A number of phenotypic properties also readily distinguished the isolate from S. griseoaurantiacus and related Streptomyces species with validly published names. It is proposed, therefore, that this organism be classified as a novel species of the genus Streptomyces, for which the name Streptomyces jietaisiensis sp. nov. is proposed. The type strain is FXJ46T (=AS 4.1859T=JCM 12279T).
162 citations
TL;DR: GmosPT showed an expression pattern similar to GiPT, the Glomus intraradices PT gene, since its transcript was more abundant in the extraradical mycelium treated with micromolar Pi levels, which opens new questions about the role and functioning of high-affinity PT in AM fungi.
Abstract: Arbuscular mycorrhizal (AM) fungi have long been shown to successfully contribute to phosphate uptake by plant roots. The first step of the fungus-mediated uptake is carried out by fungal membrane Pi transporters (PT) that transfer Pi from the soil into the extraradical hyphae. In the present work we report the identification and characterisation of a PT gene from Glomus mosseae, an AM fungus important for natural and agricultural ecosystems. Degenerate primers and rapid amplification of cDNA ends-polymerase chain reaction (PCR) allowed us to obtain a sequence (GmosPT) showing a highly significant similarity with GiPT and GvPT, the only two other PT genes already isolated from AM fungi. Reverse transcriptase-PCR experiments were carried out to study GmosPT expression profiles in structures corresponding to different fungal life stages (quiescent and germinated sporocarps, intraradical and extraradical hyphae) and in extra- and intraradical hyphae exposed to high and low Pi concentrations. GmosPT showed an expression pattern similar to GiPT, the Glomus intraradices PT gene, since its transcript was more abundant in the extraradical mycelium treated with micromolar Pi levels. In addition, the intraradical mycelium also showed a significant GmosPT expression level that was independent from external Pi concentrations. This finding opens new questions about the role and functioning of high-affinity PT in AM fungi.
156 citations
TL;DR: This is the first demonstration of variation in growth and nutrient uptake by an AMF as influenced by a localized P enrichment of the soil.
Abstract: Most terrestrial plant species form associations with arbuscular mycorrhizal fungi (AMF) that transfer soil P to the plant via their external hyphae. The distribution of nutrients in soils is typically patchy (heterogeneous) but little is known about the ability of AMF to exploit P patches in soil. This was studied by growing symbioses of Linum usitatissimum and three AMF (Glomus intraradices, G. mosseae and Gigaspora margarita) in pots with two side-arms, which were accessible to hyphae, but not to roots. Soil in one side-arm was either unamended (P0) or enriched with P; simultaneous labelling of this soil with 32 P revealed that G. intraradices responded to P enrichment both in terms of hyphal proliferation and P uptake, whereas the other AMF did not. Labelling with 33 P of P0 soil in the other side arm revealed that the increased P uptake by G. intraradices from the P-enriched patch was paralleled by decreased P uptake by other parts of the mycelium. This is the first demonstration of variation in growth and nutrient uptake by an AMF as influenced by a localized P enrichment of the soil. The results are discussed in the context of functional diversity of AMF.
TL;DR: A decrease was observed in caffeine content of the coffee pulp samples during fruiting stage, which could mean that some caffeine accumulates in the fruiting bodies of Pleurotus.
TL;DR: The data strongly suggest that the fatty acid synthase activity of arbuscular-mycorrhizal fungi is expressed exclusively in the intraradical mycelium and indicate that fatty acid metabolism may play a major role in the obligate biotrophism of arbaric fungi.
Abstract: Lipids are the major form of carbon storage in arbuscular-mycorrhizal fungi. We studied fatty acid synthesis by Glomus intraradices and Gigaspora rosea. [(14)C]Acetate and [(14)C]sucrose were incorporated into a synthetic culture medium to test fatty acid synthetic ability in germinating spores (G. intraradices and G. rosea), mycorrhized carrot roots, and extraradical fungal mycelium (G. intraradices). Germinating spores and extraradical hyphae could not synthesize 16-carbon fatty acids but could elongate and desaturate fatty acids already present. The growth stimulation of germinating spores by root exudates did not stimulate fatty acid synthesis. 16-Carbon fatty acids (16:0 and 16:1) were synthesized only by the fungi in the mycorrhized roots. Our data strongly suggest that the fatty acid synthase activity of arbuscular-mycorrhizal fungi is expressed exclusively in the intraradical mycelium and indicate that fatty acid metabolism may play a major role in the obligate biotrophism of arbuscular-mycorrhizal fungi.
TL;DR: Root exudates of Bt 176 corn significantly reduced pre-symbiotic hyphal growth, compared to Bt 11 and non-transgenic plants, suggesting that both exuded Bt toxin and defensin do not interfere with fungal host recognition mechanisms.
Abstract: We developed an experimental model system to monitor the impact of generically modified (GM) plants on arbuscular mycorrhizal (AM) fungi, a group of non-target soil microorganisms, fundamental for soil fertility and plant nutrition. The system allowed us to study the effects of root exudates of both commercial Bt corn and aubergine plants expressing Dm-AMP1 defensin on different stages of the life cycle of the AM fungal species G. mosseae. Root exudates of Bt 176 corn significantly reduced pre-symbiotic hyphal growth, compared to Bt 11 and non-transgenic plants. No differences were found in mycelial growth in the presence of Dm-AMP1 and control plant root exudates. Differential hyphal morphogenesis occurred irrespective of the plant line, suggesting that both exuded Bt toxin and defensin do not interfere with fungal host recognition mechanisms. Bt 176 affected the regular development of appressoria, 36% of which failed to produce viable infection pegs. Our experimental model system represents an easy assay for testing the impact of GM plants on non-target soil-borne AM fungi.
TL;DR: For a better understanding of the role of compounds in the AM symbiosis, studies should not be conducted only with one AM fungus looking at one fungal growth parameter such as spore germination or hyphal length, but should be wider, including several growth parameters and several AM fungi.
TL;DR: Fluorescence in situ hybridization has allowed the detection of a community of intracellular bacteria in non-axenic samples of the ectomycorrhizal fungus Laccaria bicolor S238N, and acridine orange staining suggests that the endobacteria inhabit both live and dead fungal cells.
Abstract: Fluorescence in situ hybridization, associated with confocal laser scanning microscopy or epifluorescence microscopy with deconvolution system, has allowed the detection of a community of intracellular bacteria in non-axenic samples of the ectomycorrhizal fungus Laccaria bicolor S238N. The endobacteria, mainly alpha-proteobacteria, were present in more than half of the samples, which consisted of ectomycorrhizae, fungal mats and fruit bodies, collected in the glasshouse or in the forest. Acridine orange staining suggests that the endobacteria inhabit both live and dead fungal cells. The role of these endobacteria remains to be clarified.
TL;DR: It is documented for the first time that fungal feeding soil invertebrates can significantly affect the interactions between mycorrhizal fungi and ST soil microorganisms.
Abstract: The functioning of the plant-mycorrhiza system depends on interactions with other organisms, including saprotrophic (ST) soil fungi. The interactions between mycorrhizal and ST fungi are likely affected by fungivorous soil animals, such as Collembola. In a two-factorial laboratory experiment lasting for 30 weeks we assessed the effects of an arbuscular mycorrhizal fungus (Glomus mosseae) and Collembola (Protaphorura fimata, Heteromurus nitidus and Folsomia candida) on the community composition of ST microfungi in soil planted with the invasive grass Cynodon dactylon. The presence of mycorrhiza substantially reduced total plant biomass and reduced N and P availability to the soil microflora, though these effects were less pronounced in the presence of Collembola. The density of Collembola was high (corresponding to about 2×105 individuals m−2) and was not affected by the presence of G. mosseae. In spite of the large amount of mycorrhizal mycelium in soil, it contributed little to Collembola nutrition. The presence of mycorrhiza strongly affected the community structure of ST soil fungi. In particular, mycorrhiza reduced the relative abundance of Trichoderma harzianum and Exophiala sp., but increased the abundance of Ramichloridium schulzeri and several sterile forms. However, the difference between fungal communities in mycorrhizal and non-mycorrhizal treatments was much more pronounced in the presence of Collembola. Presumably, the intense grazing by Collembola destabilized the ST fungal community, thereby making it more susceptible to the influence of G. mosseae. These results document for the first time that fungal feeding soil invertebrates can significantly affect the interactions between mycorrhizal fungi and ST soil microorganisms.
TL;DR: The results indicate that humic substances may represent a stimulatory component of the soil environment with respect to arbuscular mycorrhizal fungi.
Abstract: Effects of humic substances (humic acid or fulvic soil extract) or saprophytic microorganisms (Paecilomyces lilacinus and an unidentified actinomycete) on growth of mycelium and mycorrhiza formation by Glomus claroideum BEG23 were studied in a hydroponic system. Humic substances stimulated root colonization and production of extraradical mycelium by the mycorrhizal fungus. Both humic and fulvic acids tended to decrease populations of culturable bacteria and fungi in the cultivation system, indicating a moderately antibiotic activity. The addition of saprophytic microorganisms able to use humic substances to the cultivation system further stimulated the development of the mycorrhizal fungus. However, stimulation of G. claroideum was also observed when the saprophytic microorganisms were heat-killed, suggesting that their effect was not linked to a specific action on humic substances. The results indicate that humic substances may represent a stimulatory component of the soil environment with respect to arbuscular mycorrhizal fungi.
TL;DR: The results support the capacity of the autotrophic culture system to continuously culture arbuscular mycorrhizal fungi and may serve as a powerful tool to investigate various aspects of the symbiosis for which a source-sink relationship and photosynthetic active tissues are necessary.
Abstract: An autotrophic culture system was developed for the in vitro mycorrhization of potato plantlets. Roots of micropropagated plantlets were associated to an arbuscular mycorrhizal fungus under in vitro conditions, while shoots developed under open air conditions. Several thousand spores, an extensive extraradical mycelium and an abundant root colonization were obtained. Spores were able to colonize new plantlets under the same conditions. These results support the capacity of the autotrophic culture system to continuously culture arbuscular mycorrhizal fungi and may serve as a powerful tool to investigate various aspects of the symbiosis for which a source-sink relationship and photosynthetic active tissues are necessary.
TL;DR: Investigation of its antifungal activity clearly revealed a toxic effect on the mycelium growth of phytopathogenic fungi, such as Sclerotium rolfsii Sacc, etc., at a concentration of 80 μg recombinant CeCPI/ ml.
Abstract: A cDNA clone, designated CeCPI, encoding a novel phytocystatin was isolated from taro corms (Colocasia esculenta) using both degenerated primers/RT-PCR amplification and 5'-/3'-RACE extension. The full-length cDNA gene is 1,008 bp in size, encodes 206 amino acid residues, with a deduced molecular weight of 29 kDa. It contains a conserved reactive site motif Gln-Val-Val-Ser-Gly of cysteine protease inhibitors, and another consensus ARFAV sequence for phytocystatin. Sequence analysis revealed that CeCPI is phylogenetically closely related to Eudicots rather than to Monocots, despite taro belonging to Monocot. Recombinant GST-CeCPI fusion protein was overexpressed in Escherichia coli and its inhibitory activity against papain was identified on gelatin/SDS-PAGE. These results confirmed that recombinant CeCPI protein exhibited strong cysteine protease inhibitory activity. Investigation of its antifungal activity clearly revealed a toxic effect on the mycelium growth of phytopathogenic fungi, such as Sclerotium rolfsii Sacc. etc., at a concentration of 80 microg recombinant CeCPI/ ml. Moreover, mycelium growth was completely inhibited and the sclerotia lysed at a concentration of 150-200 microg/ml. Further studies have demonstrated that recombinant CeCPI is capable of acting against the endogenous cysteine proteinase in the fungal mycelium.
TL;DR: Results confirm that some elements required for plant growth (P, Zn, Cu) are taken up by mycorrhizal hyphae and are then transported to the plants, so that arbuscular mycor rhizal fungus colonization could partly protect plants from toxic effects of these elements.
Abstract: Mycorrhizal fungus colonization of roots may modify plant metal acquisition and tolerance. In the present study, the contribution of the extraradical mycelium of an arbuscular mycorrhizal (AM) fungus, Glomus mosseae (BEG 107), to the uptake of metal cations (Cu, Zn, Cd and Ni) by cucumber (Cucumis sativus) plants was determined. The influence of the amount of P supplied to the hyphae on the acquisition and partitioning of metal cations in the mycorrhizal plants was also investigated. Pots with three compartments were used to separate root and root-free hyphal growing zones. The shoot concentration of Cd and Ni was decreased in mycorrhizal plants compared to non-mycorrhizal plants. In contrast, shoot Zn and Cu concentrations were increased in mycorrhizal plants. High P supply to hyphae resulted in decreased root Cu concentrations and shoot Cd and Ni concentrations in mycorrhizal plants. These results confirm that some elements required for plant growth (P, Zn, Cu) are taken up by mycorrhizal hyphae and are then transported to the plants. Conversely, Cd and Ni were transported in much smaller amounts by hyphae to the plant, so that arbuscular mycorrhizal fungus colonization could partly protect plants from toxic effects of these elements. Selective uptake and transport of plant essential elements over non-essential elements by AM hyphae, increased growth of mycorrhizal plants, and metal accumulation in the root may all contribute to the successful growth of mycorrhizal plants on metal-rich substrates. These effects are stimulated when hyphae can access sufficient P in soil.
TL;DR: Differential expression of genes among the extramatrical mycelium and mycorrhizal tips indicates functional specialization of tissues forming ECM associations.
Abstract: (.)Functional compartmentation of the extramatrical mycelium of ectomycorrhizal (ECM) fungi is considered important for the operation of ECM associations, although the molecular basis is poorly characterized. (.)Global gene expression profiles of mycelium colonizing an ammonium sulphate ((NH4)(2)SO4) nutrient patch, rhizomorphs and ECM root tips of the Betula pendula-Paxillus involutus association were compared by cDNA microarray analysis. (.)The expression profiles of rhizomorphs and nutrient patch mycelium were similar to each other but distinctly different from that of mycorrhizal tips. Statistical analyses revealed 337 of 1075 fungal genes differentially regulated among these three tissues. Clusters of genes exhibiting distinct expression patterns within specific tissues were identified. Genes implicated in the glutamine synthetase/glutamate synthase (GS/GOGAT) and urea cycles, and the provision of carbon skeletons for ammonium assimilation via beta-oxidation and the glyoxylate cycle, were highly expressed in rhizomorph and nutrient patch mycelium. Genes implicated in vesicular transport, cytoskeleton organization and morphogenesis and protein degradation were also differentially expressed. (.)Differential expression of genes among the extramatrical mycelium and mycorrhizal tips indicates functional specialization of tissues forming ECM associations. (c) New Phytologist ( 2005). (Less)
TL;DR: It is shown for the first time that exploring hyphae of Aspergillus niger differentiate with respect to enzyme secretion; some strongly express the glucoamylase gene glaA, while others hardly express it at all.
Abstract: Mycelial fungi play a central role in element cycling in nature by degrading dead organic material such as wood. Fungal colonization of a substrate starts with the invasion of exploring hyphae. These hyphae secrete enzymes that convert the organic material into small molecules that can be taken up by the fungus to serve as nutrients. Using green fluorescent protein (GFP) as a reporter, we show for the first time that exploring hyphae of Aspergillus niger differentiate with respect to enzyme secretion; some strongly express the glucoamylase gene glaA, while others hardly express it at all. When a cytoplasmic GFP was used, 27% of the exploring hyphae of a 5-day-old colony belonged to the low expressing hyphae. By fusing GFP to glucoamylase and by introducing an ER retention signal, this number increased to 50%. This difference is due to cytoplasmic streaming of the reporter in the former case, as was shown by using a photo-activatable GFP. Our findings indicate that a fungal mycelium is highly differentiated, especially when taking into account that hyphae in the exploration zone were exposed to the same nutritional conditions.
01 Jan 2005
TL;DR: The enzyme tests indicate that most endophytes are degraders of the simpler sugars and cellulose available in recently dead leaves and possibly wood.
Abstract: Twenty-one endophytic isolates from Brucea javanica were tested for their ability to produce extracellular cellulase and extracellular and intracellular amylase, ligninase, pectinase and xylanase. The same fungi were tested for their ability to cause weight loss in wood blocks. All fungi produced amylase and cellulase, while only one sterile mycelium produced ligninase and no isolates produced pectinase. The enzyme tests indicate that most endophytes are degraders of the simpler sugars and cellulose available in recently dead leaves and possibly wood. Only one slow growing species of sterile mycelium however, appeared to be capable of degrading lignin that would be available in dead wood. No fungi appeared to be latent pathogens. A discussion of enzyme production in relation to possible roles of endophytes is provided.
TL;DR: Development-associated cell-death processes were investigated in detail during the growth and differentiation of Streptomyces antibioticus ATCC 11891 on confluent surface cultures, revealing the presence of a very young compartmentalized mycelium that dies following an orderly pattern, leaving alternating live and dead segments in the same hypha.
Abstract: Development-associated cell-death processes were investigated in detail during the growth and differentiation of Streptomyces antibioticus ATCC 11891 on confluent surface cultures, by using fluorescent viability probes, membrane and activity fluorescence indicators, and electron microscopy analysis. A previously unsuspected complexity was revealed, namely the presence of a very young compartmentalized mycelium that dies following an orderly pattern, leaving alternating live and dead segments in the same hypha. This death round is followed by the growth of a second mycelium which develops rapidly from the live segments of the first mycelium and dies massively in a second death round, which extends over the phases of aerial mycelium formation and sporulation.
TL;DR: The low levels of cytokines produced by A549 cells after the firm binding of either mycelium or spores of A. fumigatus may lead to insufficient recruitment of inflammatory cells to the infected site, which may result in the escape of detection by the immune defence system.
TL;DR: The successful application of EST analysis in T. harzianum is presented and a preliminary indication of gene expression in mycelium is provided, as would be expected in biocontrol fungus.
TL;DR: A number of the genes that were differentially expressed in trap cells are also known to be regulated during the development of infection structures in plant-pathogenic fungi.
Abstract: Nematode-trapping fungi enter the parasitic stage by developing specific morphological structures called traps. The global patterns of gene expression in traps and mycelium of the fungus Monacrosporium haptotylum were compared. The trap of this fungus is a unicellular spherical structure called the knob, which develops on the apex of a hyphal branch. RNA was isolated from knobs and mycelium and hybridized to a cDNA array containing probes of 2822 EST clones of M. haptotylum. Despite the fact that the knobs and mycelium were grown in the same medium, there were substantial differences in the patterns of genes expressed in the two cell types. In total, 23·3 % (657 of 2822) of the putative genes were differentially expressed in knobs versus mycelium. Several of these genes displayed sequence similarities to genes known to be involved in regulating morphogenesis and cell polarity in fungi. Among them were several putative homologues for small GTPases, such as rho1, rac1 and ras1, and a rho GDP dissociation inhibitor (rdi1). Several homologues to genes involved in stress response, protein synthesis and protein degradation, transcription, and carbon metabolism were also differentially expressed. In the last category, a glycogen phosphorylase (gph1) gene homologue, one of the most upregulated genes in the knobs as compared to mycelium, was characterized. A number of the genes that were differentially expressed in trap cells are also known to be regulated during the development of infection structures in plant-pathogenic fungi. Among them, a gas1 (mas3) gene homologue (designated gks1), which is specifically expressed in appressoria of the rice blast fungus, was characterized.
TL;DR: Results indicate that the mycelium of T. matsutake contains a novel alpha-glucan-protein complex with immunomodulatory activities that exerts modulative effects on the immune competence of mice and rats.
Abstract: Tricholoma matsutake, a high-class edible mushroom in Japan, has been reported to have excellent biological activities, but difficulty in cultivating the fruit bodies and limited bulk availability have restricted detailed studies. We have developed a method of culturing in tanks, enabling the bulk supply of the mycelia. The preparation (CM6271) exerts modulative effects on the immune competence of mice and rats. In this study, a sodium hydroxide extract of CM6271 was defatted followed by fractionation with a combination of ion exchange chromatography and gel filtration in order to identify the components involved in the expression of the activity, and a single peak fraction (MPG-1) was obtained with reversed phase chromatography. MPG-1 was a glycoprotein (sugar:protein ratio, 94.3:5.7) with a relative molecular mass of 360 kDa, and the sugar moiety contained about 90% glucose. NMR spectra and methylation analysis revealed that the α-1,4-linkage was the predominant glucan linkage with α-1,6- and α-1,2-link...
TL;DR: Cultural conditions could alter the behaviour of the fungus drastically: the outcome of the interaction between plant and fungus can be directed from mutualistic to antagonistic, characterized by fungal toxin formation and extension of the colonization to Populus shoots.
Abstract: Piriformospora indica (Sebacinaceae, Basidiomycota) is an axenically cultivable, plant growth promoting root endophyte with a wide host range, including Populus. Rooting of Populus Esch5 explants started within 6 days after transfer to WPM medium. If such plantlets with roots were inoculated with P. indica, there was an increase in root biomass, and the number of 2nd order roots was increased significantly. A totally different observation was recorded when the explants were placed into WPM with pre-grown P. indica. The interaction led to complete blocking of root production and severely inhibited plant growth. Additionally, branched aerial roots appeared which did not penetrate the medium. On contact with the fungal colony or the medium, the ends of the aerial roots became inflated. Prolonged incubation stimulated the fungus to colonize aerial parts of the plant (stem and leaves). Mycelium not only spread on the surface of the aerial parts, but also invaded the cortical tissues inter- and intracellularly. Detached Populus leaves remained vital for 4 - 5 weeks on sterile agar media or on AspM medium with pre-grown P. indica. When the fungus was pre-grown on culture media such as WPM, containing ammonium as the main source of nitrogen, leaves in contact with the cultures turned brownish within 4 - 12 h. Thereafter, the leaves bleached, and about one day later had become whitish. Thus, cultural conditions could alter the behaviour of the fungus drastically: the outcome of the interaction between plant and fungus can be directed from mutualistic to antagonistic, characterized by fungal toxin formation and extension of the colonization to Populus shoots.