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Structure-activity relationships of triterpenoid saponins on fruiting body induction in Pleurotus ostreatus.

To elucidate the structure-activity relationships of saponin on fruiting body induction in Pleurotus ostreatus, monodesmosidic saponins comprising a triterpenoid and glucose residues were synthesized. Betulin, a triterpenoid present in Birch bark, was used as the aglycon. The fruiting body induction activity increased with the number of glucose residues, from one to four. The most active betulinic saponin was betulin-3-yl beta-D-(4-O-beta-D-maltotriosyl)-glucoside.

Fruiting Bodies, Fungal↗

Localization of the stress protein SP21 in indole-induced spores, fruiting bodies, and heat-shocked cells of Stigmatella aurantiaca.

The localization and distribution of the stress protein SP21 in indole-induced vegetative cells, fruiting bodies, and heat shocked cells of Stigmatella aurantiaca were determined by immunoelectron microscopy. SP21 was found at the cell periphery in heat-shocked cells and either at the cell periphery or within the cytoplasm in indole-induced cells, often concentrated in clusters. In fruiting-body-derived spores, SP21 was located mainly at the cell wall, preferentially at the outer periphery. Furthermore, SP21 antigen was associated with cellular remnants within the stalk and within the peripheral horizon next to the fruiting body.

Bacterial Proteins↗

Guanosine metabolism and regulation of fruiting body construction in dictyostelium discoideum.

A cell aggregate of Dictyostelium discoideum either constructs a fruiting body directly or transforms into a migrating slug and fruits later on in some other locale. In the presence of formycin B, an inosine analog, and in an environment that otherwise favors fruiting, aggregates having reached a relatively late (17 hr) stage of fruit construction abandon that program and transform into migrating slugs. They then revert to the fruiting mode and construct normal fruiting bodies without further interference [Brackenbury et al. (1974) J. Mol. Biol. 90, 529-539]. The data presented here suggest that formycin B exerts its morphogenetic effect by interfering competitively with the metabolism of guanosine. Thus: see article. The recovery from formycin B is thought to result from the ensuing accumulation of guanosine and reversal of the inhibition. In support of this are the following: (1) Formycin B does cause, in vivo, an accumulation of guanosine. Exogenoug guanosine reverses the effect of formycin B, depending on their relative concentrations. (2) Guanosine is phosphorylitically cleaved to guanine and ribose-1-P by purine ribonucleoside phosphorylase (purine-nucleoside:orthophosphate ribosyl transferase, EC 2.4.2.1), present in D. discoideum extracts, and formycin B is a competitive inhibitor or this reaction with a very high affinity for the enzyme. (3) Four other analogs, also competitive inhibitors of this enzyme, produce precisely the same morphogenetic deviation. The concentrations required are consistent with the relative K1 values.

Antibiotics, Antineoplastic↗

Role of cell cohesion in Myxococcus xanthus fruiting body formation.

Dsp mutants of Myxococcus xanthus have a complex phenotype with abnormal cell cohesion, social motility, and development. All three defects are the result of a single mutation in the dsp locus, a region of DNA about 14 kilobases long. Cohesion appears to play a central role in social motility, since nonsocial mutants exhibit weak agglutination or, in the case of Dsp cells, no agglutination (L. J. Shimkets, J. Bacteriol. 166:837-841, 1986). However, Dsp cells can be agglutinated by cohesive strains of M. xanthus. This provided the opportunity to examine the role of cohesion during development by comparing the developmental phenotype of Dsp cells with that of Dsp cells mixed with cohesive strains. Dsp mutants were unable to complete any of the developmental behaviors: aggregation, fruiting body formation, developmental autolysis, and sporulation. Contact with cohesive strains seemed to restore some developmental characteristics to the Dsp cells. When allowed to develop with wild-type cells, Dsp cells accumulated in fruiting bodies and underwent developmental autolysis, but did not form a significant portion of the spore population. Igl mutants, which may be similar to the previously described frizzy mutants, are cohesive strains that are unable to form fruiting bodies. Mixing Igl cells with Dsp cells under developmental conditions resulted in fruiting body formation, although the Dsp cells were unable to form significant levels of myxospores. In spite of their inability to sporulate under developmental conditions, Dsp mutants did not appear to be defective in the sporulation process. In fact, they formed normal levels of myxospores in response to the chemical inducer glycerol.

Agglutination↗

Fine structure of fruiting bodies of Stigmatella aurantiaca (Myxobacterales).

The fruiting body of Stigmatella aurantiaca consists of a thick stalk supporting a number of individual cysts. The stalk is made of discontinuous tubules, of dimensions known for vegetative cells, which are oriented parallel to the longitudinal axis of the stalk. The red-brown cysts contain numerous, randomly oriented myxospores which are surrounded by thick, fibrous capsules. Their cell walls are wavy or ruffled and exhibit fewer budlike infoldings than reported for myxospores induced in liquid. We suggest that the extended time period available for metabolic and regulatory adjustments by the cell during morphogenesis within cysts accounts for the presence of considerably fewer deep cell wall infoldings than in glycerol-induced myxospores.

Bacteria↗

Possible involvement of Pseudomonas fluorescens and Bacillaceae in structural modifications of Tuber borchii fruit bodies.

Previous studies on Tuber borchii fruit bodies in early maturation stages suggested a role of bacteria in sporocarp structural modifications. In order to verify this hypothesis, in the present study we investigated by means of microbial and ultrastructural approaches, the bacterial population of T. borchii sporocarps from intermediate maturation phases to advanced decomposition stages, paying particular attention to chitinolytic and cellulolytic bacteria and to their relationships with ascii and ascospores. We found that Pseudomonas fluorescens and spore-forming Bacillaceae, both able to degrade cellulose and chitin, are present inside the sporocarps in all maturation stages investigated. Moreover, rod-shaped bacteria seem able to erode ascus walls and colonize the interior of ascii containing mature spores. These results suggest a possible role of these bacteria in the process of ascus opening. Moreover, the presence of P. fluorescens and Bacillaceae on isolated mature spores after decontamination suggests an intimate association between these bacteria and the ascospores.

Ascomycota↗

Constituents from the fruiting bodies of Ganoderma applanatum and their aldose reductase inhibitory activity.

Eight compounds were isolated from the fruiting bodies of Ganoderma applanatum, and were identified as 2-methoxyfatty acids (1), 5-dihydroergosterol (2), ergosterol peroxide (3) 3beta,7beta, 20,23xi-tetrahydroxy-11,15-dioxolanosta-8-en-26-oic acid (4), 7beta,20,23xi-trihydroxy-3,11,15-trioxolanosta-8-en-26-oic acid (5), cerevisterol (6), 7beta,23xi-dihydroxy-3,11,15-trioxolanosta-8,20E (22)-dien-26-oic acid (7), and 7beta-hydroxy-3,11,15,23-tetraoxolanosta-8,20E(22)-dien-26-oic acid methyl ester (8) by spectral analysis. All compounds were isolated for the first time from this fruiting bodies, and their effect on rat lens aldose reductase (RLAR) activity was tested. Among these eight compounds, ergosterol peroxide (3) was found to exhibit potent RLAR inhibition, its IC50 value being 15.4 microg/mL.

Aldehyde Reductase↗

Accumulation of trace elements in the fruiting bodies of macrofungi in the Krusné Hory Mountains Czechoslovakia.

The concentrations of some trace elements (Pb, Cd, Cu, Fe, Mn, Zn, Co, Ni) were determined in fruiting bodies of 20 fungal species from seven families (order Agaricales, Basidiomycetes) growing in the Krusné Hory Mountains, Czechoslovakia, where the air pollution is characterized as moderate. Samples were collected from three stands: a spruce forest, the waterlogged margin of a peat bog, and the peat bog itself. The biomass of fruiting bodies of all macrofungi was determined simultaneously. The trace element concentrations varied among trophic groups of fungi: saprophytic species (S) and those parasitic on Sphagnum (Sph) exhibited the highest concentrations, while wood-decomposing (Wd) species displayed the lowest. Several species mycorrhizal with spruce (Ms), such as Amanita umbrinolutea, Russula ochroleuca, and Xerocomus badius, also attained higher concentrations than were found for other mycorrhizal fungi. The trace element concentrations were higher in the caps than in the stems of the fungi. The fraction of trace metals retained by the biomass of fruiting bodies of fungi with respect to annual fallout is estimated at 1 0/00. Factors affecting fungal uptake of trace elements are discussed.

Air Pollutants↗

Occurrence of a major protein associated with fruiting body development in Neurospora and related Ascomycetes.

Electrophoretic and immunological analysis of fruiting body (perithecial) extracts demonstrates the occurrence of a major phase-specific perithecial protein in all Neurospora species and in the closely related Gelasinospora cerealis and Sordariafimicola. The perithecial proteins from these different species fall into a number of groups with different electrophoretic mobilities. They appear to be immunologically closely related but not identical to one another even within the same genus, with only partial identity exhibited between the heterothallic and pseudohomothallic Neurospora on the one hand and the homothallic Neurospora on the other hand. In immunological analysis of fruiting body extracts of the other Ascomycetes, Podospora anserina, Cochliobolus maydis, and Aspergillus nidulans, and of ascus extracts of Saccharomyces cerevisiae, no crossreaction with the Neurospora perithecial protein was found.

Ascomycota↗

Ribonucleic acid synthesis during fruiting body formation in Myxococcus xanthus.

A method has been devised that allowed us, for the first time, to pulse-label M. xanthus cells with precursors for ribonucleic acid biosynthesis while they were undergoing fruiting body formation. Using this method, we examined patterns of ribonucleic acid (RNA) accumulation throughout the process of fruiting body formation. As development proceeded, the rate of RNA accumulation increased at two periods of the developmental cycle: once just before aggregation and once late in the cycle, when sporulation was essentially completed. In contrast to vegetatively growing cells, in which only stable RNA species are labeled during a 30-min pulse, the majority of radioactivity found in RNA from 30-min pulse-labeled developing cells was found in an unstable heterodisperse fraction that migrated to the 5S to 16S region of sucrose density gradients and sodium dodecyl sulfate-polyacrylamide gels. This pattern of incorporation could not be induced (i) by a shift down of vegetatively growing cells to a nutritionally poor medium, in which the generation time was increased to that of developing cells during the growth phase, or (ii) by plating of vegetative cells onto the same solid-surface environment as that of developing cells, but which surface supported vegetative growth rather than fruiting body formation. Thus, the RNA synthesis pattern observed appeared to be related to development per se rather than to nutritional depletion or growth on a solid surface alone. The radioactivity incorporated into the unstable 5S to 16S RNA fraction accumulated as the pulse length was increased from 10 to 30 min; in contrast, an analogous unstable fraction from vegetative cells decreased as pulse length was increased. This suggested that developmental 5S to 16S RNA was more stable than vegetative cell 5S to 16S RNA (presumptive messenger RNA). However, during a 45-min chase period, radioactivity in 30-min-pulse-labeled developmental 5S to 16S RNA decayed to an extent twice that of developmental RNA located in 16S and 23S regions of sucrose density gradients and was considerably less stable than the 5S, 16S, and 23S RNA species labeled during a 30-min pulse of vegetative cells.

Micropore Filters↗

Identification, characterization, and In situ detection of a fruit-body-specific hydrophobin of Pleurotus ostreatus.

Hydrophobins are small (length, about 100 +/- 25 amino acids), cysteine-rich, hydrophobic proteins that are present in large amounts in fungal cell walls, where they form part of the outermost layer (rodlet layer); sometimes, they can also be secreted into the medium. Different hydrophobins are associated with different developmental stages of a fungus, and their biological functions include protection of the hyphae against desiccation and attack by either bacterial or fungal parasites, hyphal adherence, and the lowering of surface tension of the culture medium to permit aerial growth of the hyphae. We identified and isolated a hydrophobin (fruit body hydrophobin 1 [Fbh1]) present in fruit bodies but absent in both monokaryotic and dikaryotic mycelia of the edible mushroom Pleurotus ostreatus. In order to study the temporal and spatial expression of the fbh1 gene, we determined the N-terminal amino acid sequence of Fbh1. We also synthesized and cloned the double-stranded cDNA corresponding to the full-length mRNA of Fbh1 to use it as a probe in both Northern blot and in situ hybridization experiments. Fbh1 mRNA is detectable in specific parts of the fruit body, and it is absent in other developmental stages.

Agaricales↗

4-Acetyl-12,13-epoxyl-9-trichothecene-3,15-diol isolated from the fruiting bodies of Isariajaponica Yasuda induces apoptosis of human leukemia cells (HL-60).

The fruiting bodies of Isaria fungi have been traditionally used in Korea to treat cancer. An apoptosis-inducing compound, 4-acetyl-12,13-epoxyl-9-trichothecene-3,15-diol, was isolated from the methanol extract of fruiting bodies of Isaria japonica Yasuda by bioassay-guided fractionation. The apoptosis of the human leukemia cells (HL-60) by the compound was accessed by propidium iodide-staining flow cytometric analysis, and apoptosis-inducing activity at IC50 concentration (10 nmol/l) was further confirmed by a nuclear morphological change, a ladder pattern of internucleosomal DNA fragmentation, and an activation of caspase-3.

Apoptosis↗

An essential gene for fruiting body initiation in the basidiomycete Coprinopsis cinerea is homologous to bacterial cyclopropane fatty acid synthase genes.

The self-compatible Coprinopsis cinerea homokaryon AmutBmut produces fruiting bodies without prior mating to another strain. Early stages of fruiting body development include the dark-dependent formation of primary hyphal knots and their light-induced transition to the more compact secondary hyphal knots. The AmutBmut UV mutant 6-031 forms primary hyphal knots, but development arrests at the transition state by a recessive defect in the cfs1 gene, isolated from a cosmid library by mutant complementation. A normal primordia phenotype was achieved when cfs1+ was embedded at both sides in at least 4.0 kb of native flanking DNA. Truncations of the flanking DNA lead to reduction in transformation frequencies and faults in primordia tissue formation, suggesting that the gene is also acting at later stages of development. The cfs1 gene encodes a protein highly similar to cyclopropane fatty acid synthases, a class of enzymes shown in prokaryotes and recently in a plant to convert membrane-bound unsaturated fatty acids into cyclopropane fatty acids. In C. cinerea 6-031, the mutant cfs1 allele carries a T-to-G transversion, leading to an amino acid substitution (Y441D) in a domain suggested to be involved in the catalytic function of the protein and/or membrane interaction.

Agaricales↗

Veratryl alcohol stimulates fruiting body formation in the oyster mushroom, Pleurotus ostreatus.

The oyster mushroom, Pleurotus ostreatus, cultivated in solid state on sugarcane bagasse-wheat bran (5:1) medium in the presence of veratryl alcohol resulted in an increased production of the fruiting body at earlier times compared to when the fungus was grown in the absence of veratryl alcohol. The results indicate a new physiological role for veratryl alcohol in stimulating fruiting body formation. Veratryl alcohol also stimulated laccase production during the mycelial growth stage. Evidence is also presented that laccases were involved in the physiological development of the fruiting body.

Benzyl Alcohols↗

A sigma(54) activator protein necessary for spore differentiation within the fruiting body of Myxococcus xanthus.

Insertion of an internal DNA fragment into the act1 gene, which encodes one of several sigma(54)-activator proteins in Myxococcus xanthus, produced a mutant defective in fruiting body development. While fruiting-body aggregation appears normal in the mutant, it fails to sporulate (<10(-6) the wild-type number of viable spores). The A and C intercellular signals, which are required for sporulation, are produced by the mutant. But, while it produces A-factor at levels as high as that of the wild type, the mutant produces much less C-signal than normal, as measured either by C-factor bioassay or by the total amount of C-factor protein detected with specific antibody. Expression of three C-factor-dependent reporters is altered in the mutant: the level of expression of Omega4414 is about 15% of normal, and Omega4459 and Omega4403 have alterations in their time course. Finally, the methylation of FrzCD protein is below normal in the mutant. It is proposed that Act1 protein responds to C-signal reception by increasing the expression of the csgA gene. This C-signal-dependent increase constitutes a positive feedback in the wild type. The act1 mutant, unable to raise the level of csgA expression, carries out only those developmental steps for which a low level of C-signaling is adequate.

Adaptor Proteins, Signal Transducing↗

Isolation of transcripts preferentially expressed during fruit body primordia differentiation in the basidiomycete Agrocybe aegerita.

An Agrocybe aegerita cDNA library, constructed from fruit body primordia poly(A)+ RNAs, was screened by differential colony hybridization. Clones which preferentially hybridized to poly(A)+ RNA sequences from fruit body primordia, versus poly(A)+ RNAs from mycelium, were isolated. Eight of these clones (EMAa-1 to EMAa-8) encoded eight different poly(A)+ RNAs which were demonstrated to be undetectable in the four stages preceding primordia formation and to be concomitantly accumulated when primordia differentiate, suggesting that EMAa gene products are closely involved in the morphogenesis of primordia. The eight EMAa cDNAs hybridize to at least seven unique regions distributed randomly in the A. aegerita genome. The expression of two EMAa cDNA sequences in E. coli led to the isolation of their gene products as fusion proteins.

Basidiomycota↗

137Cs content in the fruit bodies of various Tuber species.

In this research, the concentration of 137Cs in the fruit bodies of the Tuber species T. magnatum Pico, T. borchii Vitt., T. aestivum Vitt., and T. excavatum Vitt. collected in three different regions of Italy was determined. The values obtained have been compared to the soil concentration of 137Cs, and the transfer factor was determined. The radiocesium content of the examined fruit bodies ranged from 2.5 Bq kg(-1) to 33.3 Bq kg(-1) fresh weight; the median transfer factor values of the four species ranged between 0.06 and 0.6. Our findings indicate that the radiocesium level in truffles from these regions of Italy is generally low, and, thus, their consumption is not of radiological concern. The results may suggest certain hypotheses as to the mechanisms involved in radiocesium uptake in these fungi.

Ascomycota↗

[Lipid composition of fruiting bodies and submerged mycelium from Lentinus edodes (Berk.) Sing (Lentinula edodes (Berk.) Peglar)].

Comparison of Lentinus edodes with true xylotrophs and humus saprotrophs cultivated under the conditions used in commercial fungal production revealed the following differences in the lipid composition of the fruiting bodies: (1) in L. edodes, fatty acids are characterized by a high C16:0 level and the presence of C16:1; (2) neutral lipids of L. edodes have a high monoglyceride and free fatty acid content and a low triglyceride level; and (3) the main L. edodes phospholipids are phosphatidylethanolamine and cardiolipin. Fruiting bodies of L. edodes differ from its submerged mycelium in lipid quantity and composition. Submerged mycelium contains up to 20% lipids (fruiting bodies contain 3-4%), neutral lipids are predominantly composed of triglycerides and free fatty acids, and the main constituents of phospholipids are phosphatidylcholine and cardiolipin.

Fatty Acids↗