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A three-dimensional model of myxobacterial fruiting-body formation.

Myxobacterial cells are social; they swarm by gliding on surfaces as they feed cooperatively. When they sense starvation, tens of thousands of cells change their movement pattern from outward spreading to inward concentration and form aggregates that become fruiting bodies. Cells inside fruiting bodies differentiate into round, nonmotile, environmentally resistant spores. Traditionally, cell aggregation has been considered to imply chemotaxis, a long-range cell interaction that shares many features of chemical reaction-diffusion dynamics. The biological evidence, however, suggests that Myxococcus xanthus aggregation is the consequence of direct cell-contact interactions that are different from chemotaxis. To test whether local interactions suffice to explain the formation of fruiting bodies and the differentiation of spores within them, we have simulated the process. In this article, we present a unified 3D model that reproduces in one continuous simulation all the stages of fruiting-body formation that have been experimentally observed: nonsymmetric initial aggregates (traffic jams), streams, formation of toroidal aggregates, hemispherical 3D mounds, and finally sporulation within the fruiting body.

Cell Communication↗

Chemical analysis of the lamella walls of Agaricus bisporus fruit bodies.

Purified lamella wall fragments of Agaricus bisporus fruit bodies were analyzed and shown to consist of neutral sugars (46.5%), hexosamines (31.7%), proteins (9.5%), some lipid material (10.0%), and ash (1.4%). The cell walls were fractionated on the basis of their polysaccharide solubility in water and alkaline solutions. The isolated fractions, using methylation analysis, exhibited striking chemical structural differences compared with the same fractions obtained from the corresponding vegetative cells and fruit bodies (stipe and pileus) walls. The structural differences detected in the wall seem to correspond to the ultimate differentiation of the mycelium inside the fruit body of A. bisporus.

Agaricus↗

Genetic evidence for somatic haploidization in developing fruit bodies of Armillaria tabescens.

Armillaria spp. have vegetative hyphae with diploid uninucleate cells, but the fruit bodies of many species contain clamped dikaryotic hyphae. Earlier observations suggest that somatic haploidization takes place in developing fruit bodies. To verify this, a uninucleate diploid cell was isolated from each of the 49 mating combinations between single-spore isolates of Armillaria tabescens and they were fruited. Twenty-four isolates produced fruit bodies with at least a partially dikaryotic subhymenium. Dikaryotic hyphae were isolated from fruit-body primordia and homokaryons were obtained by micromanipulation or by protoplasting. Approximately half of the isolates proved to represent recombinant mating types in respect to parent homokaryons, and most of them contained recombinant haploid DNA, based on random-amplified microsatellite markers. The results show that the nuclei in dikaryotic hyphae found in fruit bodies result from somatic haploidization. The mechanism of haploidization remains unclear.

Basidiomycota↗

Scanning electron microscopy of fruiting body formation by myxobacteria.

Scanning electron microscopy was used to follow fruiting body formation by pure cultures of Chondromyces crocatus M38 and Stigmatella aurantica. Vegetative cells were grown on SP agar and then transferred to Bonner salts agar for fructification. Fruiting in both species commences with the formation of aggregation centers which resemble a fried egg in appearance. In Chondromyces the elevated center or "yolk" region of the aggregation enlarges into a bulbous structure under which the stalk forms and lengthens. At maximum stalk height the bulb extends laterally as bud-like swellings appear. These are immature sporangia and are arranged in a distintive radial pattern around the top of the stalk. This symmetry is lost as more sporangia are formed. Stigmatella does not form a bulb; rather the yolk region of the aggregation center projects upward to form a column-like stalk which is nearly uniform in diameter throughout its length. At maximum stalk height, the terminus of the stalk develops an irregular pattern of bud-like swellings. These differentiate into sporangia. Stalks of 2-week-old mature fruiting bodies of both species appear to be cellular in composition. Stereomicrographs suggest orientation of these cells parallel to the long axis of the stalk. Stalks of 8-week-old fruiting bodies of Chondromyces were acellular and consisted of empty tubules, suggesting that the cells undergo degeneration with aging of the fruiting body.

Microscopy, Electron, Scanning↗

[Trypsin-like proteinases and trypsin inhibitors in fruiting bodies of higher fungi].

The activity of trypsin-like proteinases and trypsin inhibitors was measured in fruiting bodies of various species of basidial fungi (Basidiomycetes). Fruiting bodies of all fungi contained these enzymes, with the exceptions of polypore (Coriolus versicolor (Fr.) Karst) and hedgehog fungus (Hericium erinaceus (Fr.) Quel), belonging to the families Polyporaceae and Hericiaceae, respectively, in which the enzyme activities were barely detectable. The activity of trypsin-like proteinases was the highest in fruiting bodies of Boletaceae and Agaricaceae. Fruiting bodies of all fungi contained trypsin inhibitors. The highest activity of trypsin inhibitors was detected in basidiomycetes of the families Boletaceae, Agaricaceae, and Pleurotaceae, including Boletus castanus (Fr.) Karst, orange-cap boletus (Leccinum aurantiacum (Fr.) Sing), and brown-cap boletus (Leccinum melanum (Fr.) Karst).

Basidiomycota↗

Isolation and characterization of differentially expressed genes in the mycelium and fruit body of Tuber borchii.

The transition from vegetative mycelium to fruit body in truffles requires differentiation processes which lead to edible fruit bodies (ascomata) consisting of different cell and tissue types. The identification of genes differentially expressed during these developmental processes can contribute greatly to a better understanding of truffle morphogenesis. A cDNA library was constructed from vegetative mycelium RNAs of the white truffle Tuber borchii, and 214 cDNAs were sequenced. Up to 58% of the expressed sequence tags corresponded to known genes. The majority of the identified sequences represented housekeeping proteins, i.e., proteins involved in gene or protein expression, cell wall formation, primary and secondary metabolism, and signaling pathways. We screened 171 arrayed cDNAs by using cDNA probes constructed from mRNAs of vegetative mycelium and ascomata to identify fruit body-regulated genes. Comparisons of signals from vegetative mycelium and fruit bodies bearing 15 or 70% mature spores revealed significant differences in the expression levels for up to 33% of the investigated genes. The expression levels for six highly regulated genes were confirmed by RNA blot analyses. The expression of glutamine synthetase, 5-aminolevulinic acid synthetase, isocitrate lyase, thioredoxin, glucan 1,3-beta-glucosidase, and UDP-glucose:sterol glucosyl transferase was highly up-regulated, suggesting that amino acid biosynthesis, the glyoxylate cycle pathway, and cell wall synthesis are strikingly altered during morphogenesis.

Ascomycota↗

Influence of the substrate on the ultrastructure of Pleurotus pulmonarius fruit body primordia.

Pleurotus pulmonarius fruit body primordia (FBP) formed on potato extract agar (PEA) and wheat straw-based media (WS) were studied. When grown on PEA, FBP hyphae had a large number of vacuoles and less stainable cytoplasmic material. In contrast, when grown on WS, there was abundant cytoplasmic material and fewer vacuoles. FBP grown on WS had a more well-defined mushroom shape than those grown on PEA. The average hyphal diameters on PEA and WS were 4.15 and 3.52 micro m, respectively; the average hyphal lengths were 11.75 and 11.80 micro m, respectively. The control mechanisms regulating formation of the normal shape and size of FBP might be more dependent on the amount of cytoplasmic material than on the physical volume of the hyphal compartment. Since the ultrastructure of the hyphal compartment, which makes up the substance of the FBP, depends on the substrate in which the FBP has been grown, the composition of the substrate may be important for manipulating the nutritive and organoleptic characteristics of the mushroom crop.

Hyphae↗

C-factor: a cell-cell signaling protein required for fruiting body morphogenesis of M. xanthus.

During fruiting body development, the product of the csgA gene is necessary for cellular aggregation, for spore differentiation, and for gene expression that is initiated after 6 hr of starvation. From nascent wild-type fruiting bodies we have purified a polypeptide of 17 kd called C-factor, which, at approximately 1 to 2 nM, restores normal development to csgA mutant cells. C-factor activity is not recovered from extracts of unstarved, growing cells or csgA mutant cells. The amino acid sequence from purified C-factor demonstrates that it is the product of the csgA gene. C-factor is active over a narrow range of concentration and has properties of a morphogenetic paracrine signal.

Amino Acid Sequence↗

FibA and PilA act cooperatively during fruiting body formation of Myxococcus xanthus.

The extracellular matrix (ECM) of Myxococcus xanthus is essential for social (S-) motility and fruiting body formation. An ECM-bound protein, FibA, is homologous to M4 zinc metalloproteases and is important for stimulation by a phosphatidylethanolamine (PE) chemoattractant and for formation of discrete aggregation foci. In this work, we demonstrate that a correlation exists between a reduced ability to respond to PE and the observed defects in fruiting body morphogenesis. Furthermore, the fibA aggregation defect is accentuated by the absence of either PilA, the structural subunit of type IV pili, or DifD, a chemosensory response regulator. The inability to form fruiting bodies is not due to a loss of S-motility, but rather the loss of PilA and pili as pilT fibA mutants form fruiting bodies. The FibA active site residue E342 is important for fruiting body morphogenesis in the absence of PilA. Mutants exhibiting defects in fruiting body morphogenesis also produce fewer viable spores. It is proposed that FibA and PilA act as extracellular sensors for developmental signals.

Amino Acid Sequence↗

The anti-hyperglycemic activity of the fruiting body of Cordyceps in diabetic rats induced by nicotinamide and streptozotocin.

Little scientific evidence exists to support the numerous herbs used to improve diabetes-related metabolic disorders. Cordyceps, a Chinese herbal medicine with fruiting body and carcass, has been proposed to have multiple medicinal activities. The objective of this study was to investigate the effects of fruiting body and carcass of Cordyceps on hyperglycemia. Male Wistar rats administered with placebo (STZ group), 1 g of fruiting body (FB group), 1 g of carcass (CC group), or 1g of fruiting body plus carcass (CF group) of Cordyceps for four weeks (d1 to d28) were injected with nicotinamide (200 mg/kg) and streptozotocin (65 mg/kg) on d15. Animals fed with placebo and injected with saline acted as the controls (CON group). The results showed that water intake (d15 to d29), changes in fasting blood glucose concentration (d15 to d26), and serum concentrations of fructosamine (d29) were significantly greater in the STZ, CC and CF groups than in the CON and FB groups (one-way ANOVA, P < 0.05). The diabetic rats had significantly lower weight gain and higher blood glucose response in oral glucose tolerance test than the control rats; and these changes were significantly reduced by administrating the fruiting body of Cordyceps. Our results revealed that fruiting body, not carcass, of Cordyceps attenuated the diabetes-induced weight loss, polydipsia and hyperglycemia, and these improvements suggest that fruiting body of Cordyceps has a potential to be the functional food for diabetes.

Administration, Oral↗

Isolation of genes differentially expressed during the fruit body development of Pleurotus ostreatus by differential display of RAPD.

To analyze genes involved in fruit body development of Pleurotus ostreatus, mRNAs from three different developmental stages: i.e., vegetative mycelium, primordium, and mature fruit body, were isolated and reverse-transcribed to cDNAs. One hundred and twenty random PCR amplifications were performed with the cDNAs, which generated 382, 394, 393 cDNA fragments from each developmental stage. From these fragments, four cDNA clones specifically expressed in primordium or mature fruit body were detected. Sequence analysis and database searches revealed significant similarity with triacylglycerol lipase, cytochrome P450 sterol 14 alpha-demethylase and developmentally regulated genes of other fungi. Northern blot analyses confirmed that all of the four cDNAs were unexpressed in mycelium, thus stage-specific genes for fruit body formation of P. ostreatus were successfully isolated.

Fruiting Bodies, Fungal↗

[Studies on the morphology of fruit-body developing of Polyporus umbellatus].

OBJECTIVE: Studying the fruit-body development of polyporus umbellatus to reveal the life-cycle of this medicinal fungus. METHODS: Parts of materials treated by paraffin section, and examined under a light microscope; parts of materials by ultrathin section and observation under transmission electron microscope (TEM). RESULTS: The fruit-body consists of trimitic hyphae, including generative hyphae, skeletal hyphae and binding hyphae. Generative hyphae are characterized by thin walls, dense cytoplasmic contents. Skeletal hyphae are unbranched, thick-walled, and of a narrow lumen. Binding hyphae are much-branched, tending to weave themselves among the other hyphase of flesh. The morphology of basidiospore development was also discussed in this paper. CONCLUSIONS: The trimitic hyphae are the main component parts in the different developing stages of fruit-body of polyporus umbellatus.

Plants, Medicinal↗

Trace elements in fruiting bodies of ectomycorrhizal fungi growing in Scots pine (Pinus sylvestris L.) stands in Poland.

The trace metal contents in fruiting bodies of ectomycorrhizal (ECM) fungi, symbiotic partners of Scots pine, were studied on three sites situated in west-central Poland. Elements were determined by atomic absorption spectrometry in 123 samples of 16 species. The study explored the differences in metal accumulation in relation to site, fungal species, age and part of the fruiting body and results were related to metal content in soil and plant material (roots and needles). Soil analysis revealed that results were obtained under environmental conditions not subject to strong anthropogenic pressure. Median metal concentrations did not differ disparately between sites, although the concentrations of each of the tested metals in the individual species varied to a large extent. Extremely high levels of Al with a large bioconcentration factor (BCF) were found in sporocarps of Thelephora terrestris. The spread between the highest and the lowest concentration (max/min) was very wide in Al, Cd and Pb and these elements may be considered to be absorbed preferentially by fruiting bodies of some species whereas Fe, Mn and Zn, with relatively low values of max/min, are normally absorbed by the majority of fungi. There was no clear relationship between caps and stipes in metal content. However, a tendency to higher metal concentration in the caps was observed. The metal content in young and older fruiting bodies of five different fungi was species dependent. In order to estimate the degree of accumulation of each element by plant and mushrooms, bioconcentration factors (BCFs) were calculated. In plant material (roots and needles), highest values of BCFs were noted for essential metals, like Zn and Mn. Lead showed a definite exclusion pattern (BCF below 1). In fruiting bodies of tested fungi, especially in Amanita muscaria, cadmium was the most intensively accumulated metal. Lead was excluded by plants but was accumulated or excluded by fungi depending on the species. The significance of mycological observation and quantification of the metal content in monitoring of the forest ecosystem is discussed.

Age Factors↗

Dual genetic loci and flavonoid metabolism orchestrate fruiting body coloration in Flammulina filiformis: a multi-omic roadmap for fungal pigmentation.

BACKGROUND: The fruiting bodies of macrofungi exhibit diverse coloration, traditionally attributed to melanin and carotenoid biosynthesis. This study is the first to reveal that flavonoids, rather than these classical pigments, are the predominant contributors to yellow pigmentation in the Flammulina filiformis. OBJECTIVE: To uncover the genetic basis and key regulatory genes involved in pigment formation in F. filiformis fruiting bodies, and to establish a model framework for studying color genetics in macrofungi. METHODS: Metabolomic profiling was conducted on yellow and white F. filiformis fruiting bodies to identify key pigment components. A segregating population was constructed, followed by integrated multi-omics analyses-including bulk segregant analysis (BSA), genome-wide association study (GWAS), and transcriptomics-to map regulatory loci and candidate genes. Functional roles were validated via genetic transformation and protein structural modeling. RESULTS: Flavonoid accumulation was identified as the biochemical hallmark of pigmented fruiting bodies. Genetic analysis revealed a dual regulatory mechanism: a qualitative locus governing pigmentation presence and a quantitative trait determining color intensity. Combined BSA and GWAS pinpointed a major locus, Ffcrs, within a recombination-suppressed region. Transcriptomic analysis identified two key regulators, Ffakr (a transcriptional activator) and Ffpal (encoding phenylalanine ammonia-lyase). Functional verification via transformation, structural modeling, and metabolite profiling in transgenic lines confirmed their essential roles in flavonoid biosynthesis and pigmentation. CONCLUSION: This study uncovers a flavonoid-based pigmentation mechanism in F. filiformis and elucidates a complex genetic architecture shaped by both qualitative and quantitative loci, providing a new paradigm for understanding pigment formation in macrofungi. The identified regulatory factors establish a molecular foundation for the precise manipulation of economically important pigmentation traits in edible mushroom.

Flavonoids↗

The effect of light on fruiting body formation and adenosine 3':5'-cyclic monophosphate metabolism in Coprinus macrorhizus.

The monokaryotic mycelia of a mutant strain, fis(c), of Coprinus macrorhizus, which are able to form monokaryotic fruiting bodies in the light, failed to form any fruiting bodies in darkness. A dikaryon and a mutant strain, ds, formed malformed fruiting bodies in darkness. Illumination for 1 day of fis(c) mycelia grown in darkness for 4 days or longer was effective in inducing malformed fruiting bodies. The accumulation of adenosine 3':5'-cyclic monophosphate in the illuminated mycelia of strain fis(c) was demonstrated. The illuminated mycelia of strain fis(c) produced high levels of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] and phosphodiesterase (3':5'-cyclic-AMP 5'-nucleotidohydrolase, EC 3.1.4.17), which degrades cAMP, while the dark-grown mycelia showed no or very low activities of these enzymes. Dikaryotic mycelia and monokaryotic mycelia of strain ds produced significant amounts of these enzymes even in darkness. When the dark-grown mycelia of strain fis(c) were exposed to continuous light, the activities of adenylate cyclase and phosphodiesterase increased rapidly after a lag period whose length depends on the culture age of mycelia. Cycloheximide inhibited the increase in these enzyme activities stimulated by light. When the fis(c) mycelia were exposed to continuous light, an increase in cAMP-binding activity was observed. A possible participation of cAMP in the formation of fruiting bodies in C. macrorhizus is discussed.

Adenylyl Cyclases↗

Structural characterization of beta-glucans of Agaricus brasiliensis in different stages of fruiting body maturity and their use in nutraceutical products.

beta-Glucans of Agaricus brasiliensis fruiting bodies in different stages of maturity were isolated and characterized by FTIR and NMR. These fractions had greater amount of (1-->6)-beta-glucan and the (1-->3)-beta-glucan increased with fruiting bodies maturation. Yields of beta-glucans increased from 42 mg beta-glucans g(-1) fruiting bodies (dry wt) in immature stage to 43 mg g(-1) in mature stage with immature spores, and decreased to 40 mg g(-1) in mature stage with spore maturation. Mature fruiting bodies, which included these glucans, have potential therapeutical benefits for use in nutraceutical products.

Agaricus↗

Cucurbitane triterpenes from the fruiting bodies and cultivated mycelia of Leucopaxillus gentianeus.

A reinvestigation of the fruiting bodies of the mushroom Leucopaxillus gentianeus, allowed the isolation of two minor cucurbitane triterpenes, namely, cucurbitacin D (5) and the new metabolite 16-deoxycucurbitacin B (6). The latter compound lacks an oxygenated substituent at C-16, an unprecedented structural feature among congeners of cucurbitacin B. The cucurbitanes present in the fruiting bodies were compared with those extracted from mycelia grown on the modified Melin-Norkans (MMN) culture medium. Cucurbitacins B (1) and D (5), as well as leucopaxillones A (3) and B (4), were isolated from both sources; in contrast, 16-deoxycucurbitacin B (6) and a mixture of fatty acid esters of cucurbitacin B (2) were absent in the mycelia. A new triterpene, 18-deoxyleucopaxillone A (7), was isolated from the mycelia, but was not detected in the fruiting bodies. The antiproliferative activity of the isolated triterpenes was determined against the NCI-H460 human tumor cell line, in comparison with the antitumor compound topotecan, a well-known topoisomerase I inhibitor.

Agaricales↗

The gene for a lectin-like protein is transcriptionally activated during sexual development, but is not essential for fruiting body formation in the filamentous fungus Sordaria macrospora.

BACKGROUND: The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies called perithecia that protect the developing ascospores and ensure their proper discharge. In previous microarray analyses, several genes have been identified that are downregulated in sterile mutants compared to the wild type. Among these genes was tap1 (transcript associated with perithecial development), a gene encoding a putative lectin homolog. RESULTS: Analysis of tap1 transcript levels in the wild type under conditions allowing only vegetative growth compared to conditions that lead to fruiting body development showed that tap1 is not only downregulated in developmental mutants but is also upregulated in the wild type during fruiting body development. We have cloned and sequenced a 3.2 kb fragment of genomic DNA containing the tap1 open reading frame and adjoining sequences. The genomic region comprising tap1 is syntenic to its homologous region in the closely related filamentous fungus Neurospora crassa. To determine whether tap1 is involved in fruiting body development in S. macrospora, a knockout construct was generated in which the tap1 open reading frame was replaced by the hygromycin B resistance gene hph under the control of fungal regulatory regions. Transformation of the S. macrospora wild type with this construct resulted in a tap1 deletion strain where tap1 had been replaced by the hph cassette. The knockout strain displayed no phenotypic differences under conditions of vegetative growth and sexual development when compared to the wild type. Double mutants carrying the Deltatap1 allele in several developmental mutant backgrounds were phenotypically similar to the corresponding developmental mutant strains. CONCLUSION: The tap1 transcript is strongly upregulated during sexual development in S. macrospora; however, analysis of a tap1 knockout strain shows that tap1 is not essential for fruiting body formation in S. macrospora.

Fungal Proteins↗