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PubMed2003

Fingerprint profiling of acid hydrolyzates of polysaccharides extracted from the fruiting bodies and spores of Lingzhi by high-performance thin-layer chromatography.

Modern extraction and planar chromatographic instrumentation were employed for the fingerprint profiling of carbohydrates from an important and popular medicinal mushroom commonly known as Lingzhi. For the first time, the feasibility of employing the high-performance thin-layer chromatography (HPTLC) peak profiles (fingerprints) of carbohydrates for the screening of various Lingzhi species/products was demonstrated. An analytical procedure was developed such that upon acid hydrolysis of the polysaccharides extracted from various Lingzhi samples, fingerprint profiles that reveal the relative amounts of the degradation products, such as mono- and oligosaccharides, can be obtained using HPTLC plates (Si 50000) for separation and 4-aminobenzoic acid as the post-chromatographic derivatization reagent for detection. Also, using automated multiple development (AMD), the acid hydrolyzates from Lingzhi, consisting of simple and more complex sugars, can be separated simultaneously with high degree of automation. An important finding was that unique fingerprint patterns were observed in the monosaccharide profiles between two highly valued Lingzhi species, Ganoderma applanatum and Ganoderma lucidum, under total or partial acid hydrolysis conditions. Additionally, the HPTLC fingerprint profiles of carbohydrates were obtained from the extracts of the spores and fruiting bodies of Lingzhi and compared.

Acids↗
PubMed1991

Targeted disruption of the Myxococcus xanthus orotidine 5'-monophosphate decarboxylase gene: effects on growth and fruiting-body development.

The Myxococcus xanthus gene coding for orotidine 5'-monophosphate (OMP) decarboxylase (EC 4.1.1.23) was cloned. The M. xanthus uraA gene efficiently complemented an Escherichia coli OMP decarboxylase mutant, permitting it to grow in the absence of uracil. Electroporation of M. xanthus with a circular plasmid carrying a selectable uraA::kan gene disruption resulted in homologous recombination at the chromosomal uraA locus. Chromosomal integration of the gene disruption plasmid created heterozygous (uraA+/uraA::kan) tandem duplications. These tandem duplications were unstable and segregated auxotrophic uraA::kan daughters at frequencies of 2 x 10(-4) to 8 x 10(-4) per viable cell. Rare uraA::kan segregants were easily obtained by selecting for resistance to the toxic analog 5-fluoroorotic acid. Our experiments suggest that the cloned uraA gene could facilitate the use of gene duplications in the genetic analysis of M. xanthus development. The uraA mutants could utilize uracil, uridine, or uridine 5'-phosphate for growth, indicating that M. xanthus has pyrimidine salvage pathways. During multicellular development, uraA::kan gene disruption mutants sporulated to wild-type levels but formed smaller and more numerous aggregates than did their uraA+ parent, regardless of whether uracil was added to the medium. Pyrimidine deprivation of uraA mutants, under conditions that otherwise supported vegetative growth, failed to induce fruiting-body development or sporulation.

Cloning, Molecular↗
PubMed2004

Promoter analysis of cgl2, a galectin encoding gene transcribed during fruiting body formation in Coprinopsis cinerea (Coprinus cinereus).

In the homobasidiomycete Coprinopsis cinerea, expression of the two fruiting body-specific galectins, CGL1 and CGL2, is controlled by nutrients, light and darkness and the A mating type genes. In this study, we analyzed the promoter of the cgl2 gene by measuring transcript levels by quantitative real-time PCR and show that regulation of CGL2 expression occurs at the transcriptional level. A minimal promoter sufficient to confer regulated expression of a heterologous reporter gene and comprising 627 base pairs from the start codon was defined. On the minimal promoter we identified a 120 bp sequence mediating induction of the cgl2 gene in constant darkness. Along with direct repeats (TGGAAG/TGGAAG/GGAA), the sequence contains a CRE consensus site (cAMP-responsive element, TGCGTCA) suggesting the involvement of cAMP signaling in cgl2 activation. No specific elements responsible for light repression and mating type regulation were found in the promoter.

Adaptation, Physiological↗
PubMed2005

Ganodermin, an antifungal protein from fruiting bodies of the medicinal mushroom Ganoderma lucidum.

A 15-kDa antifungal protein, designated ganodermin, was isolated from the medical mushroom Ganoderma lucidum. The isolation procedure utilized chromatography on DEAE-cellulose, Affi-gel blue gel, CM-Sepharose and Superdex 75. Ganodermin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and CM-Sepharose. Ganodermin inhibited the mycelial growth of Botrytis cinerea, Fusarium oxysporum and Physalospora piricola with an IC50 value of 15.2 microM, 12.4 microM and 18.1 microM, respectively. It was devoid of hemagglutinating, deoxyribonuclease, ribonuclease and protease inhibitory activities.

Amino Acid Sequence↗
PubMed2006

Mushrooms by magic: making connections between signal transduction and fruiting body development in the basidiomycete fungus Schizophyllum commune.

The basidiomycete fungus Schizophyllum commune has been utilised as a model system for examining the genetic regulation of sexual reproduction, a process that culminates in the production of mushrooms in this and many other related species. Recent studies have suggested that conserved elements of the cyclic AMP (cAMP) signalling pathway play a role in the control of mushroom development in S. commune. The small G-protein Ras also appears to impinge on the process, either by inputting into the cAMP pathway, or by acting in parallel. The molecular connections between nutrient sensing and mushroom development are now beginning to be examined.

Adaptation, Physiological↗
PubMed2006

Macrophage-stimulating activity of polysaccharides extracted from fruiting bodies of Coriolus versicolor (Turkey Tail Mushroom).

The macrophage-stimulating effect of polysaccharides extracted from Coriolus versicolor (Turkey Tail mushroom) was investigated, and their effectiveness was compared with that of lipopolysaccharide (LPS). The purified polysaccharide (CV-S2-Fr.I) of C. versicolor obtained by Sepharose CL-6B gel chromatography stimulated macrophage lysosomal enzyme activity by 250% at a concentration of 100 microg/mL, which was higher than that of LPS at the same concentration. When CV-S2-Fr.I was used in combination with interferon-gamma, there was a marked cooperative induction of nitric oxide production. However, CV-S2-Fr.I had no effect on nitric oxide production by itself. The proportion of C3-positive macrophages in the CV-S2-Fr.I group increased by 7.2-fold compared with the control group.

Agaricales↗
PubMed2000

Cloning and characterization of two hydrophobin genes differentially expressed during fruit body development in Lentinula edodes.

Hydrophobins play important roles in morphogenesis and pathogenesis in fungi and fruit development in mushrooms. Two genes encoding hydrophobins (Le.hyd1 and Le.hyd2) were isolated during sequencing of random clones from a primordial cDNA library of Lentinula edodes. The nucleotide sequences of these two genes were determined. These two genes are 760 and 738 bp in length and the deduced amino acid sequences are homologous to various fungal hydrophobins with characteristic cysteine spacing. These hydrophobin genes are Class I hydrophobins judging by their conserved domains and hydropathy patterns. The transcript level of Le.hyd1 is high in primordium and that of Le.hyd2 is high in dikaryotic mycelial tissues. Poor expression of these two genes in monokaryotic parents indicates that these two genes are under mating-type regulation. We thus suggest that differential expression of these two L. edodes hydrophobins during fruit development may contribute to their distinct roles in fruiting of this mushroom.

Amino Acid Sequence↗
PubMed1983

Evidence for long-lived mRNA during fruiting body formation in myxococcus xanthus.

Half-lives of Myxococcus xanthus mRNA were determined by inhibiting RNA polymerase with rifampin and then measuring the rate of [35S]methionine incorporation into protein. Vegetative cells resuspended in clone fruiting liquid culture showed an average mRNA half-life of approximately 3.5 min. Developmental cells (24 or 48 hr) exhibited biphasic decay curves with apparent mRNA half-lives of approximately 3.5 min and 20-30 min. The more-stable mRNA species accounted for about 30% of all mRNA present in 24-hr cells and as much as 40% of all mRNA of 48-hr cells. Analysis of the 35S-labeled proteins by NaDodSO4/polyacrylamide gel electrophoresis showed that only 5-10 major polypeptides are synthesized after 30 min of rifampin treatment. One of these is protein S, the spore surface coat protein, because immunoprecipitation of the 35S-labeled proteins with antisera specific for protein S showed continued synthesis of protein S in the presence of rifampin. The half-life for its mRNA was calculated to be 15-30 min. RNA from vegetative and developing cells was pulse labeled with 32PO4 followed by rifampin treatment. Analysis of the labeled RNA on 3% NaDodSO4/polyacrylamide gels showed 5-10 long-lived mRNA bands in developing cells. These results show that there are several abundant mRNA species synthesized developmentally that are exceptionally long lived. The fact that the majority of the mRNA species show the shorter half-life suggests that developing cells retain the normal mechanism for mRNA degradation.

Bacterial Proteins↗
PubMed1983

The organisation of fruiting body formation in Dictyostelium minutum.

The process of culmination was investigated in three strains of the species Dictyostelium minutum. After aggregates have been formed a pulsatile signalling mechanism arises; the centre of signal emission becomes the apex of the developing fruiting structure. In the late aggregate, all cells differentiate into prespore cells. Cells that have reached the apex of the culminating cells mass redifferentiate into stalk cells. In two of the three D. minutum strains, interruption of regular stalk formation, more or less random formation of stalk cells and the synthesis of stalk supporting material from cell debris often takes place. The formation of multiple apices on aggregates and early fruiting structures is characteristic for these two strains. Within the species D. minutum, the exhibition of a marked pulsatile signalling mechanism is correlated with a capacity to form a regularly shaped stalk and to organize relatively large cell masses. The possible function of pulsatile signalling in the culmination process is discussed.

Cell Differentiation↗