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PubMed1994

Intraspecific molecular variation among Trichoderma harzianum isolates colonizing mushroom compost in the British Isles.

The genetic diversity in Trichoderma harzianum isolates from mushroom compost was assessed using various molecular techniques. Restriction fragment length polymorphism (RFLP) analysis of ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) divided the 81 isolates into three major groups, 1, 2 and 3. There was no variation within a group in rDNA, while a low degree of polymorphism was detected in mtDNA. Random amplified polymorphic DNA (RAPD) analysis of 30 randomly chosen isolates, with six primers, in general confirmed the RFLP groups. Nucleotide sequence determination of rDNA internal transcribed spacer (ITS) 1 revealed three distinct ITS types, 1, 2 and 3, possessed by isolates from the respective groups 1, 2 and 3. Based on these molecular data, group 2 isolates, which are aggressive colonizers of mushroom compost, could be clearly distinguished from the isolates belonging to the other two groups.

Base Sequence↗

PubMed1997

Isolation of developmentally regulated genes from the edible mushroom Agaricus bisporus.

From a cDNA library, constructed from mushroom primordia, nine cDNAs were isolated which were either induced or specifically expressed during fruit body development and maturation of the basidiomycete Agaricus bisporus. These cDNAs varied in size from 372 to 1019 bp and hybridized to transcripts of 400-1600 nt. Four of the cDNAs were only expressed in the generative phase of the life cycle while the other five cDNAs were strongly induced but had low steady-state mRNA levels in vegetatively grown mycelium of the hybrid strain Horst U1. An apparent full-length cDNA could be identified by sequence analysis and specified a putative protein homologous to the delta-subunit of the mitochondrial ATP synthase complex of Saccharomyces cerevisiae and Neurospora crassa. For one of the partial cDNAs, significant homology was found with a family of cell division control proteins, while another partial cDNA appeared to encode a cytochrome P450. All cDNAs, except the presumed cytochrome-P450-specifying cDNA (cypA), hybridized with single copy genes scattered over the Agaricus genome. For the cypA gene, the presence of several additional copies was shown by heterologous hybridizations. Based on changes in expression levels of the fruit-body-induced genes during development coinciding with alterations in morphological appearance of mushrooms, four stages of development were distinguished during growth and maturation of A. bisporus fruit bodies.

Agaricus↗

PubMed2002

Fine-scale genetic analyses reveal unexpected spatial-temporal heterogeneity in two natural populations of the commercial mushroom Agaricus bisporus.

This study examined the fine-scale genetic variation of the commercial mushroom, Agaricus bisporus, over 2 years at two sites in France. One site was a meadow fertilized with horse manure and disturbed regularly by humans; the other was a Monterey cypress forest free of human disturbance. Altogether, 50 mushrooms were collected and analysed for mitochondrial and nuclear genetic variation marked by RFLPs and multilocus enzyme electrophoretic polymorphisms. Population samples from these two sites were genetically different and both sites contained high levels of genetic diversity. No identical genotypes were found at either site between the 2 years and there was little evidence for extensive vegetative clonality for this species. Contrary to expectations, very limited evidence of pseudohomothallic reproduction was found. Results from tests of Hardy-Weinberg equilibrium and genotypic equilibrium showed that outcrossing and recombination have played significant roles in both populations. The results demonstrated spatial-temporal genetic heterogeneity of A. bisporus in natural populations.

Agaricus↗

PubMed1999

Different temporal and spatial expression of two hydrophobin-encoding genes of the edible mushroom Agaricus bisporus.

In a search for genes that are only expressed in fruit bodies of the basidiomycete Agaricus bisporus, two cDNAs, hypA and hypB that encode hydrophobins have been isolated previously. In this study, the structure of hypB is resolved and it is shown that the two genes are differentially expressed, indicating that the encoded hydrophobins serve different functions in A. bisporus mushrooms. hypB encodes a polypeptide (HYPB) of 119 aa that shows little sequence identity with HYPA apart from the characteristic arrangement of eight cysteines found exclusively in hydrophobins. The temporal and spatial expression of the two hydrophobin-encoding genes during fruit body development was compared using Northern analysis and in situ hybridization. Accumulation of hypA mRNA was found in tissue fractions consisting of undifferentiated white hyphae. In situ hybridization showed that the highest hypA mRNA levels are not found in the outermost cell layers of the pileipellis but in the cell layers adjacent to that. The highest level of expression of hypB occurs early in development when the primordium differentiates into densely packed, randomly oriented cap hyphae and loosely packed, vertically oriented stipe hyphae. In mature mushrooms, a strong accumulation of hypB transcripts was found only in the transitional zone between cap and stipe tissue, demonstrating that transcription regulation of hypB is clearly distinct from hypA.

Agaricus↗

PubMedDate not supplied

Vision affects mushroom bodies and central complex in Drosophila melanogaster.

The brain of Drosophila is structurally altered by sensory stimuli that the flies receive during their adult life. Size and fiber number of the mushroom bodies, central complex, and optic lobes are influenced by social, spatial, or olfactory cues. Recently, the optic lobes have been shown to depend on the light regime that flies experience. Structural plasticity in the brain is thought to be a correlate of functional adaptations and long-term memory. We therefore extend our investigation of volume changes to the calyces of the mushroom bodies and the central complex. We show that rearing flies in constant light for 4 days increases the volume of both structures by up to 15% compared to rearing them in total darkness. Much of this difference develops during the first day. The effect of light is not hormonally mediated, as monocularly deprived flies develop a smaller ipsilateral calyx. Mutant analysis suggests that light generates its effects through known visual pathways. In contrast to the optic lobes, in the calyx and central complex structural changes can be linked to cAMP signaling, as in the mutants dunce1 and amnesiac1 no volume differences are observed. Surprisingly, the mutant rutabaga1 shows a prominent light-dependent volume increase in the calyx and central complex, dissociating structural from behavioral plasticity. In complete darkness wild-type flies grow larger calyces under crowded conditions in their normal culture vials than if kept in small groups on fresh food. This stimulating effect of crowding is not observed in any of the cAMP mutants, including rutabaga1.

Aging↗

PubMed1993

Characterization of a Lectin from Lactarius deterrimus (Research on the Possible Involvement of the Fungal Lectin in Recognition between Mushroom and Spruce during the Early Stages of Mycorrhizae Formation).

A lectin (LDetL) was isolated from carpophores of the mushroom Lactarius deterrimus, a specific symbiont of the spruce, by a combination of affinity, hydroxylapatite, and gel-filtration chromatography. Its molecular mass, as determined by gel filtration, is about 37,000 D, and its structure is dimeric, with two identical subunits assembled by noncovalent bonds. It appeared homogeneous on high-performance liquid chromatography gel filtration, but isoelectric focusing revealed microheterogeneity, with a main band in the pH zone near 6.5. Amino acid analysis showed that LDetL contains a large proportion of glycine and especially methionine. Hapten inhibition assay indicated that LDetL is most specific for [beta]-D-galactosyl(1->3)-D-N-acetyl galactosamine residues. The lectin was formed in the in vitro-cultivated mycelium, and anti-lectin antibodies revealed by indirect immunofluorescence the presence of lectin in the cell wall. Receptor sites for LDetL were found on the roots, especially on the root hairs, of axenically grown spruce seedlings. The lectin LDL previously isolated by us from the taxonomically related mushroom Lactarius deliciosus, a symbiont of the pine, does not bind to the spruce radicle. This suggests a role of the fungal lectin in recognition and specificity during the early stages of mycorrhizae formation.

Journal Article↗

PubMed1978

Biochemical Induction of Fruiting in Schizophyllum: Isolation and Preliminary Purification of an Inducing Substance from Agaricus bisporus Mushrooms.

A diffusible substance was isolated from Agaricus bisporus mushrooms that induced haploid fruiting body formation in Schizophyllum commune. An efficient isolation procedure was developed. The fruiting-inducing substance was shown to be present in all stages of Agaricus mushroom development and there was no preferential accumulation of the activity in gill, cap, or stipe tissues. Some chemical and physical properties of the inducing factor are described.

Journal Article↗

PubMed2006

Adjuvant effect of mushroom glucan and bovine lactoferrin upon Aeromonas hydrophila vaccination in catla, Catla catla (Hamilton).

Mushroom glucan and bovine lactoferrin (Lf), known for their immunostimulatory potential, were used as adjuvant in conjunction with a formalin-killed Aeromonas hydrophila vaccine in catla, Catla catla. In vitro antigen-specific responsiveness of catla leucocytes and protective responses against experimental challenge with homologous antigen were monitored following immunization. Antigen-specific proliferation, 'macrophage activating factor' (MAF) production and antibody production were significantly higher in fish injected with glucan adjuvanted vaccine. Lf adjuvanted preparations showed a weak proliferative response and MAF production, although the antibody production was significantly higher than the controls. A good degree of protection was achieved with the glucan adjuvanted vaccine. However, in spite of producing significant anti-A. hydrophila antibody, Lf adjuvanted vaccine did not confer any protection following challenge with A. hydrophila. The potential of adjuvanticity of mushroom glucan and bovine Lf in intraperitoneal vaccination is discussed.

Adjuvants, Immunologic↗

PubMed1993

Hallucinogenic mushroom use by Danish students: pattern of consumption.

A survey among Danish students revealed that 33 (7.2%) of the respondents had experience with hallucinogenic mushrooms (HM). Afterwards these students answered an extended questionnaire about habits and pattern of consumption. It seems that mushroom use is experimental and may be due to risk-seeking behaviour but is mostly recreational and restricted to a few events. Age over 25 years and male sex were statistically related to a higher lifetime consumption rate of HM. The age of first time users was significantly lower for marijuana than for HM. Only 15 of the respondents wanted to continue use. Most of them were males and they had significantly more friends with HM experience than those who would not continue. We recommend that surveys of drug abuse include questions concerning HM to monitor the trends and extent of HM abuse.

Adolescent↗

PubMed1997

Ingestive and inhalative allergy to the mushroom Boletus edulis.

This is the first report on inhalative and ingestive allergy to the common edible mushroom Boletus edulis (Be) (English, edible boletus; German, Steinpilz; French, bolet; Italian, porcino or boleto) belonging to the class Basidiomycetes. Four cases observed in our allergy unit are presented, showing different clinical manifestations of this rare allergy: as an occupational problem or life-threatening anaphylactic reactions after eating Be. In all cases, skin prick-to-prick tests with raw Be were strongly positive: in three cases, specific IgE against Be could be found. The symptoms were reproducible after an inhalation challenge test. It is noteworthy that not only can Basidiomycetes cause airborne allergy but also that edible mushrooms from this class can cause inhalative and intestinal allergy. The two patients with strong anaphylactic reactions demonstrate that Be may have great allergenic potential.

Adult↗

PubMed2001

Use of molecular markers to differentiate between commercial strains of the button mushroom Agaricus bisporus.

Agaricus bisporus is an edible basidiomycete cultivated industrially for food production. Different spawn and mushroom producers use genetically related A. bisporus strains frequently marketed as different products. In this paper we show that the use of suitable molecular markers reveals the high level of genetic homology of commercial strains of A. bisporus, and allows, at the same time, to distinguish between them. In the course of this work, a molecular marker potentially linked to the agronomic character 'mushroom weight' has been identified by bulked segregant analysis.

Agaricus↗

PubMedDate not supplied

pH-dependent interconvertible forms of mushroom tyrosinase with different kinetic properties.

The lag in cresolase activity and inhibition by excess tyrosine of mushroom tyrosinase which was observed when assayed at pH 6.8 was found to be absent when assayed at pH 5.0. The absence of lag and inhibition by excess tyrosine of tyrosinase at pH 5.0 were brought about only after the enzyme was kept at pH 5.0, at 0-4 degrees C, for 1.5 h. The enzyme kept at pH 5.0 for 1.5-3 h at 0-4 degrees C when brought back to pH 6.8, acquires lag and inhibition by excess tyrosine when its activity was measured at pH 6.8. The pH-dependent changes in the kinetic properties of the mushroom tyrosinase are similar to the pH-dependent changes in the kinetic properties of tyrosinase from B-16 murine melanoma and human skin, and thus appear to be a general property of tyrosinase from diverse sources.

Basidiomycota↗

PubMed1995

Complex formation between mushroom tyrosinase and Manduca dopachrome isomerase.

Melanin biosynthesis in animals is initiated by the ubiquitously present tyrosinase and is aided by dopachrome isomerase. We have characterized a novel dopachrome isomerase (decarboxylating) from the hemolymph of Manduca sexta that generates a new quinone methide intermediate during melanogenesis (Sugumaran, M. and Semensi, V. (1991) J. Biol. Chem. 266, 6073-6078). This enzyme has the ability to form a complex with mushroom tyrosinase as judged by a number of physicochemical studies. The isomerase exhibited a marked inhibitory effect on tyrosinase and tyrosinase reciprocated by inhibiting the isomerase. While the isomerase showed no activity toward preformed dopaminechrome, it readily influenced the stability of dopaminechrome generated in situ by tyrosinase. Moreover, mushroom tyrosinase, which lacked specific binding to Concanavalin A Sepharose column, after complexing with the isomerase exhibited binding to this column. The complex formation also affected the pI value as well as mobility on a size exclusion column of these enzymes. Enzymes executing sequential metabolic transformation are known to form complexes called metabolons. Based on these above studies, it is concluded that both the enzymes involved in insect melanogenic pathway--phenoloxidase and dopachrome isomerase--are able to form a metabolon complex.

Animals↗

PubMed1976

Delayed release nutrient supplement for mushroom culture.

The disadvantages associated with the supplementation of noncomposted nutrients to mushroom compost at spawning were largely overcome by encapsulating microdroplets of vegetable oil within a protein coat that was denatured with formaldehyde. Increases in mushroom yield of 60% were obtained. Delayed nutrient release was indicated by prolonged stimulation of yields beyond the first few flushes.

Journal Article↗

PubMed1991

Evolution of Volatile Sulfur Compounds during Laboratory-Scale Incubations and Indoor Preparation of Compost Used as a Substrate in Mushroom Cultivation.

Volatile sulfur compounds are known to be produced during the preparation of compost used as a substrate in mushroom cultivation. Because they cause odor problems, attempts have been made to reduce the production of these compounds. The influences of temperature and various additions on the production of volatile sulfur compounds from composting material were tested on laboratory-scale preparations. The production of H(2)S, COS, CH(3)SH, and (CH(3))(2)S was proven to be a biological process with an optimal temperature that coincides with the optimal temperature for biological activity. The formation of CS(2) and (CH(3))(2)S(2) was shown to be a nonbiological process. The emission of volatile sulfur compounds during the indoor preparation of mushroom compost appeared to be remarkably reduced (about 90%) as compared with the emission during the conventional outdoor process. Introduction of this indoor composting process would result in a significant reduction in environmental pollution.

Journal Article↗

PubMed1992

Growth Characteristics of the Thermophilic Fungus Scytalidium thermophilum in Relation to Production of Mushroom Compost.

Scytalidium thermophilum is an important thermophilic fungus in the production of mushroom compost. I investigated the characteristics of this organism and present a simple model with which fungal growth in compost can be described. The model is used to predict better circumstances for rapid indoor production of mushroom compost. I conclude that inoculation of the starting material with prepared compost either before or after the pasteurization phase has only a minor effect on the shortening of the composting process. This is because the initial growth rate of the fungus is much higher than its growth rate later. A lower temperature (53.5 degrees C instead of the usual 56 to 58 degrees C) during the pasteurization phase may be most profitable for rapid compost production; such a temperature may reduce the time that is needed for the last phase of the production process by at least 1 day.

Journal Article↗

PubMed1994

Inheritance of Strain Instability (Sectoring) in the Commercial Button Mushroom, Agaricus bisporus.

The button mushroom, Agaricus bisporus, is a commercially important cultivated filamentous fungus. During the last decade, the button mushroom industry has depended mainly on two strains (or derivatives of these two strains). Using one of these highly successful strains (strain U1) we examined the phenomenon of strain instability, specifically, the production of irreversible sectors. Three "stromatal" and three "fluffy" sectors were compared with a healthy type U1 strain and with a wild-collected isolate. Compost colonization and fruit body morphology were examined. The main objective of this study, however, was to examine the meiotic stability of the sectored phenotype. Single basidiospores were isolated and subjected to a grain bioassay in which the ability to produce sectors was measured. Our results were as follows: (i) basidiospore cultures obtained from a wild-collected isolate showed no tendency to produce sectors; (ii) approximately 5% of the basidiospore cultures obtained from healthy type U1 strains produced irreversible sectors in the grain bioassay; (iii) the five primary sectors examined produced basidiospore cultures, half of which produced normal-looking growth in the grain bioassay and half of which produced some degree of sectoring; and (iv) the one sectored isolate that represented the F2 generation gave ratios similar to the 1:1 ratio observed for the F1 cultures.

Journal Article↗

PubMed1998

Morphological and molecular identification of Trichoderma isolates on North American mushroom farms.

Green mold disease (causal agent, Trichoderma) has resulted in severe crop losses on mushroom farms worldwide in recent years. We analyzed 160 isolates of Trichoderma from mushroom farms for morphological, cultural, and molecular characteristics and classified these isolates into phenotypic groups. The most common group comprised approximately 40% of the isolates and was identified as a strain of Trichoderma harzianum. This group was consistently recovered from farms with severe green mold disease but not from farms with little or no problem. In addition, the strain identified as the major cause of green mold disease in Ireland and the United Kingdom grouped with these North American isolates in having very similar randomly amplified polymorphic DNA patterns.

Basidiomycota↗