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Genetic variation and joint segregation of biochemical loci in the common meadow mushroom, Agaricus campestris.

In this paper we report the results of an electrophoretic examination of genetic variation in the common meadow mushroom, Agaricus campestris. An examination of variability at 10 biochemical loci in a single natural population revealed an average heterozygosity of 0.281. Mendelian segregation was verified for 13 loci in single-spore-derived lines from individual sporophores. One linkage was found (Gpt-2 with Pep-GL-1, r = 0.065) among 47 pairwise examinations for joint segregation.

Basidiomycota↗

[Lead in mushrooms (author's transl)].

The lead content of 222 species of wild mushrooms, either edible or poisonous, was determined by graphite furnace atomic absorption spectroscopy. Altogether 494 samples, grown in rural regions of southern Germany, were tested. The lead content was between less than 2.0 and 40.4, on the average 10.6 mg/kg dry weight, equivalent to less than 0.1 and 3.0, on the average 1.06 mg/kg fresh weight.

Basidiomycota↗

[Crude, digestible and indigestible fruit body proteins in oyster mushroom Pleurotus ostreatus].

Different kinds of protein (crude, digestible, non-digestible) were analysed in caps and stipe of Pleurotus ostreatus (oyster mushroom) in four phases of development. The phases examined were: a (cap diameter < 5 cm); B (diameter 5-8 cm); C (diameter, 8-10 cm) and D (diameter > 10 cm). The Pleurotus variety analysed (the cap and the stipe) has a relative high crude protein content, the main part of which is digestible (average, 92%). During the four stages of fruit body ripening, stage B was the best, with the highest crude and digestible protein concentrations. In stage D the highest non-digestible protein content was measured in the cap and stipe. These data can be used for optimal harvesting of fruit bodies of cultivated P. ostreatus.

Digestion↗

[Determination of cadmium binding proteins in the edible mushroom (Agaricus arvensis Schff, ex Fr.) (author's transl)].

The isolation and characterization of cadmium compounds from Agaricus arvensis Schff. ex Fr. are described. These compounds were extractable with buffers. Ultrafiltrations and protein precipitations suggested protein bound cadmium. The mushroom proteins were separated by gel chromatography and the cadmium content in the corresponding fractions was determined by flameless atomic absorption. Two cadmium containing protein fractions were found with cadmium concentrations of 2.1 and 1 mg/g protein, respectively. The molecular weights of both cadmium containing proteins were estimated to be about 2,000 and 15,000--20,000, respectively by comparison with standard proteins.

Agaricales↗

Analysis of 2,4-D and 2,4,5-T in lingonberries, wild mushrooms, birch and aspen foliage.

A method was developed for residue analysis of 2,4-D and 2,4,5-T in lingonberries, wild mushrooms, birch and aspen foliage. The method involves the extraction of the residues with diethyl ether followed by alkali hydrolysis, purification by column chromatography, methylation of the free acids and their determination by gas chromatography. The limit of detection is 0.05 ppm for 2,4-D and 0.02 ppm for 2,4,5-T for a 20 g sample with a recovery of 75% +/- 11%. Results of some analyses are also reported.

2,4,5-Trichlorophenoxyacetic Acid↗

[Increased levels of cesium-137 and cesium-134 in 34 mushroom species following the reactor accident at Chernobyl].

From September to December of 1986, 85 samples of 34 species of wild mushrooms were collected, mostly in northern Baden and occasionally in neighboring areas. Cs-137, Cs-134, and the natural nuclide K-40 were determined by gamma-spectrometry. Samples from the Rhine Valley and the Kraichgau were generally much less contaminated than those from the Black Forest. With one exception, their radiocesium levels were below the Common Market limit of 600 Bq/kg. The exception was a sample of Laccaria amethystina with 1710 Bq/kg Cs-137 and 766 Bq/kg Cs-134. Relatively high levels were also found in Xerocomus badius, X. chrysenteron and Cantharellus tubaeformis. The hymenophore contained higher concentrations of radiocesium than the other parts of the body of the fruit. No correlation between the concentration of K-40 and that of the Cs nuclides was found. The ratio of Cs-137:Cs-134 was usually below 3 but reached the value of 11.8 in Piptoporus betulinus growing on birch wood. In this case, the substrate must have contained more radiocesium from the time before the Chernobyl reactor accident than from the time thereafter.

Accidents↗

Amaninamide, a new toxin of Amanita virosa mushrooms.

Amaninamide, a toxin closely related to the family of amatoxins, was found exclusively in Amanita virosa mushrooms. It differs from the well known toxin alpha-amanitin in that it lacks the 6'-hydroxyl group of the tryptophan unit, and from the toxin amanin found in Amanita phalloides by the presence of a carboxamide group instead of a carboxylic acid groups.

Agaricales↗

Evolution of mushroom mitochondrial DNA: Suillus and related genera.

Mapping studies were performed with 18 cloned probes on mitochondrial DNA (mtDNA) from 15 species of Suillus and four species from three related genera of fleshy pore mushrooms (Boletaceae). Within Suillus, mtDNAs vary in size from 36 to 121 kb, differ in gene order by only one major rearrangement, and have diverged in nucleotide sequence within the large subunit ribosomal RNA gene region by up to 2.9%. Three additional gene orders exist in related genera. Two of the three can be transformed into the predominant Suillus order by either one or two rearrangements. The fourth requires two to three rearrangements to be converted to any of the others. The minimum estimates of nucleotide divergence within the large subunit ribosomal RNA gene region vary from 8.3% to 11% in comparisons between Suillus and these related species. Trees based on restriction-site and size differences within the mitochondrial ribosomal RNA genes were consistent with the hypothesized sequence of genome rearrangements and provide suggestive evidence for a major expansion of the mitochondrial genome within Suillus. Structural and sequence changes in mtDNA provided information about phylogenetic relationships within the Boletaceae.

Basidiomycota↗

Transformation of the cultivated mushroom, Agaricus bisporus, to hygromycin B resistance.

Application of biotechnology to the cultivated mushroom, Agaricus bisporus, has been hampered thus far by the lack of a transformation system. Here, transformation of both a homo- and a heterokaryotic strain of A. bisporus to hygromycin B resistance is described. Transforming DNA was integrated into the A. bisporus genome and stably maintained throughout vegetative growth. Transformants of the heterokaryotic strain formed transgenic fruiting bodies. Promoters derived from the unrelated ascomycete Aspergillus nidulans and from A. bisporus itself, were able to drive expression of the hygromycin B resistance gene. Expression controlled by a fragment of 265 bp from the A. bisporus GPD promoter was sufficient to generate transformants. However, transformation efficiency was not enhanced by using this homologous promoter.

Agaricus↗

The effect of some osmolytes on the activity and stability of mushroom tyrosinase.

The thermodynamical stability and remained activity of mushroom tyrosinase (MT) from Agaricus bisporus in 10 mM phosphate buffer, pH 6.8, stored at two temperatures of 4 and 40 degrees C were investigated in the presence of three different amino acids (His, Phe and Asp) and also trehalose as osmolytes, for comparing with the results obtained in the absence of any additive. Kinetics of inactivation obey the first order law. Inactivation rate constant (kinact) value is the best parameter describing effect of osmolytes on kinetic stability of the enzyme. Trehalose and His have the smallest value of kinact (0.7x10(-4) s-1) in comparison with their absence (2.5x10(-4) s-1). Moreover, to obtain effect of these four osmolytes on thermodynamical stability of the enzyme, protein denaturation by dodecyl trimethylammonium bromide (DTAB) and thermal scanning was investigated. Sigmoidal denaturation curves were analysed according to the two states model of Pace theory to find the Gibbs free energy change of denaturation process in aqueous solution at room temperature, as a very good thermodynamic criterion indicating stability of the protein. Although His, Phe and Asp induced constriction of MT tertiary structure, its secondary structure had not any change and the result was a chemical and thermal stabilization of MT. The enzyme shows a proper coincidence of thermodynamic and structural changes with the presence of trehalose. Thus, among the four osmolytes, trehalose is an exceptional protein stabilizer.

Agaricus↗

Mycelium growth kinetics and optimal temperature conditions for the cultivation of edible mushroom species on lignocellulosic substrates.

The influence of environmental parameters on mycelial linear growth of Pleurotus ostreatus, P. eryngii, P. pulmonarius, Agrocybe aegerita, Lentinula edodes, Volvariella volvacea and Auricularia auricula-judae was determined in two different nutrient media in a wide range of temperature, forming the basis for the assessment of their temperature optima. V. volvacea grew faster at 35 degrees C, P. eryngii at 25 degrees C, P. ostreatus and P. pulmonarius at 30 degrees C, A. aegerita at 25 or 30 degrees C and A. auricula-judae at 20 or 25 degrees C depending on the nutrient medium used and L. edodes at 20 or 30 degrees C depending on the strain examined. The mycelium extension rates were evaluated on seven mushroom cultivation substrates: wheat straw, cotton gin-trash, peanut shells, poplar sawdust, oak sawdust, corn cobs and olive press-cake. The mycelium extension rates (linear growth and colonization rates) were determined by the 'race-tube' technique, and were found to be the highest on cotton gin-trash, peanut shells and poplar sawdust for Pleurotus spp. and A. aegerita. Wheat straw, peanut shells and particularly cotton gin-trash supported fast growth of V. volvacea, whereas wheat straw was the most suitable substrate for L. edodes and A. auricula-judae. Supplemented oak sawdust and olive press-cake were poor substrates for most species examined, while almost all strains performed adequately on corn cobs.

Agaricales↗

Antioxidant and memory enhancing effects of purple sweet potato anthocyanin and cordyceps mushroom extract.

The effects of purple sweet potato anthocyanin (SPA) and Cordyceps mushroom extract (CME) on lipid peroxidation, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and cognitive deficits were examined. Both SPA and CME exhibited DPPH radical scavenging activities with similar potency. In contrast, only SPA was shown to effectively inhibit lipid peroxidation initiated by Fe2+ and ascorbic acid in rat brain homogenates. Furthermore, SPA markedly enhanced cognitive performance, assessed by passive avoidance test in ethanol-treated mice. Combined treatments with SPA and CME did not significantly influence the effects of SPA alone. These results demonstrate that anthocyanin prepared from purple sweet potato exhibits memory enhancing effects, which may be associated with its antioxidant properties.

Administration, Oral↗

The hepatocellular bile acid transporter Ntcp facilitates uptake of the lethal mushroom toxin alpha-amanitin.

Hepatotoxicity caused by the mushroom poison alpha-amanitin is an unusual but serious cause of death and liver transplantation. Understanding the mechanisms of alpha-amanitin uptake may lead to rational therapeutic approaches. Because older data suggested that a sodium-dependent bile acid transporter is responsible for alpha-amanitin uptake, we tested the hypothesis that Na(+)-taurocholate cotransporter polypeptide (Ntcp) facilitates hepatocellular alpha-amanitin uptake. Human hepatoblastoma cells (HepG2), cells that have lost native Ntcp expression, were stably transfected with the rat Ntcp gene. Taurocholate uptake by the transfected cells exhibited a physiologically normal K(m) and V(max). A toxicologically relevant functional assay for alpha-amanitin uptake was developed by measuring its ability to block cytokine-induced synthesis of interleukin-1 receptor antagonist (IL-1Ra) mRNA. Treatment with interleukin-1beta (10 ng/ml) and interleukin-6 (100 ng/ml) increased IL-1Ra mRNA abundance 8.6-fold and 15.6-fold in HepG2 cells and Ntcp-transfected cells, respectively. Pretreatment of transfected cells with 1 micro M alpha-amanitin for 6-10 h almost completely blocked induction of IL-1Ra mRNA (1.9-fold induction) whereas pretreatment of non-transfected cells did not block induction of IL-1Ra mRNA (21.6-fold induction, P<0.02 compared with stimulated transfected cells without alpha-amanitin). These findings demonstrate that Ntcp may be an important mediator of alpha-amanitin uptake by the liver.

Amanitins↗

Influence of spawn rate and commercial delayed release nutrient levels on Pleurotus cornucopiae (oyster mushroom) yield, size, and time to production.

Pleurotus cornucopiae 608 was grown on a mixture of pasteurized cottonseed hulls (75% dry wt). 24% chopped wheat straw, and 1% ground limestone. The substrate was spawned at various levels (1.25%, 2.5%, 3.75%, or 5% wet wt) and not supplemented or supplemented with commercial delayed release nutrient (Campbell's S-41) at various levels (0%, 3%, 6%, 9%, or 12%). Maximum yield (weight of fresh mushrooms harvested at maturity) was obtained at 3.75-5% spawn level and 6% S-41 supplement. As supplement levels exceeded 6%, yields declined significantly. There was a negative correlation (r=-0.81) between spawn rate and days to production. As the spawn rate increased, the number of days to production decreased. By using a spawn rate of 3.75% of the wet substrate wt, it was possible to reduce the time to production by a mean of 9.2 days compared with a spawn rate of 1.25%.

Crops, Agricultural↗

Laccase from the medicinal mushroom Agaricus blazei: production, purification and characterization.

The medicinal mushroom Agaricus blazei produced high amounts of laccase (up to 5,000 units l(-1)) in a complex, agitated liquid medium based on tomato juice, while only traces of the enzyme (<100 units l(-1)) were detected in synthetic glucose-based medium. Purification of the enzyme required three chromatographic steps, including anion and cation exchanging. A. blazei laccase was expressed as a single protein with a molecular mass of 66 kDa and an isoelectric point of 4.0. Spectroscopic analysis of the purified enzyme confirmed that it belongs to the "blue copper oxidases". The enzyme's pH optimum for 2,6-dimethoxyphenol (DMP) and syringaldazine was pH 5.5; but for 2,2'-azino-bis(3-ethylthiazoline-6-sulfonate) (ABTS) no distinct pH optimum was observed (highest activity at the lowest pH tested). Purified laccase was stable at 20 degrees C, pH 7.0 and pH 3.0, but rapidly lost its activity at 40 degrees C or pH 10. Sodium chloride strongly inhibited the enzyme activity, although the inhibition was completely reversible. The following kinetic constants were determined (K(m), k(cat)): 63 microM, 21 s(-1) for ABTS, 4 microM, 5 s(-1) for syringaldazine, 1,026 microM, 15 s(-1) for DMP and 4307 microM, 159 s(-1) for guaiacol. The results show that--in addition to the wood-colonizing white-rot fungi--the typical litter-decomposing basidiomycetes can also produce high titers of laccase in suitable liquid media.

Agaricus↗

Purification of a laccase from fruiting bodies of the mushroom Pleurotus eryngii.

A purification scheme involving ion exchange chromatography on diethylaminoethyl (DEAE) cellulose, carboxymethyl (CM) cellulose, and Q-Sepharose, followed by fast protein liquid chromatography-gel filtration on Superdex 75, was utilized to isolate a laccase from the fruiting bodies of the mushroom Pleurotus eryngii. The laccase was adsorbed on DEAE-cellulose and Q-Sepharose and unadsorbed on CM-cellulose. The molecular mass of the laccase in sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 34 kDa. The laccase activity was maximal at 70 degrees C and remained high at 80 degrees C. High activity of the laccase was maintained at ph 3 to 5. The activity underwent an abrupt decline at pH 6 and 7 and was indiscernible at ph 8 and 9. The laccase exhibited an inhibitory activity toward HIV-1 reverse transcriptase with an IC50 of 2.2 microM. Its N-terminal sequence was dissimilar to those of laccase isoenzymes previously isolated from cultured mycelia of the same species.

Amino Acid Sequence↗