Bisporus Strains Research Articles

Cultivo do champignon em função da temperatura

O cogumelo Agaricus bisporus é um cogumelo de clima temperado que requer baixa temperatura para a indução da frutificação. Essa exigência limita o seu cultivo no Brasil nos meses mais frios do ano ou exige a utilização de ambiente controlado, onerando muito a produção. Por isso, este trabalho teve como objetivo avaliar a produtividade de diferentes linhagens em função da temperatura de cultivo após a indução da frutificação. Diferentes linhagens de A. bisporus foram avaliadas quanto ao efeito da temperatura sobre a colonização do composto e a produtividade do cogumelo. As temperaturas de 21 e 25°C foram utilizadas durante a colonização do composto e, após a indução da frutificação, durante o ciclo de cultivo. Para todas as linhagens testadas, a temperatura de 25°C proporcionou maior velocidade de colonização do composto, favorecendo ciclos de cultivo mais curtos. Além disso, após a indução da frutificação a 18°C, a manutenção do ambiente de cultivo a 25°C proporcionou maior produtividade, quando comparada ao cultivo a 21°C. Para a linhagem mais produtiva, os três primeiros fluxos foram responsáveis por mais de 50% da produção total. Portanto, para um cultivo comercial em que é necessário manter volumes constantes de produção, os ciclos de cultivo poderiam ser limitados a três fluxos de produção.

Dynamics of the chemical composition and productivity of composts for the cultivation of Agaricus bisporus strains

Two compost formulations based on oat straw (Avena sativa) and brachiaria (Brachiaria sp.) were tested for the cultivation of three Agaricus bisporus strains (ABI-07/06, ABI-05/03, and PB-1). The experimental design was a 2 × 3 factorial scheme (composts × strains) with 6 treatments and 8 repetitions (boxes containing 12 kg of compost). The chemical characterization of the compost (humidity, organic matter, carbon, nitrogen, pH, raw protein, ethereal extract, fibers, ash, cellulose, hemicellulose, and lignin) before and after the cultivation of A. bisporus and the production (basidiomata mass, productivity, and biological efficiency) were evaluated. Data were submitted to variance analysis, and averages were compared by means of the Tukey's test. According to the results obtained, the chemical and production characteristics showed that the best performances for the cultivation of A. bisporus were presented by the compost based on oat and the strain ABI-07/06.

Regulation of three genes encoding cell-wall-degrading enzymes ofTrichoderma aggressivumduring interaction withAgaricus bisporus

Members of the genus Trichoderma are very effective competitors of a variety of fungi. Cell-wall-degrading enzymes, including proteinases, glucanases, and chitinases, are commonly secreted as part of the competitive process. Trichoderma aggressivum is the causative agent of green mould disease of the button mushroom, Agaricus bisporus. The structures of 3 T. aggressivum genes, prb1 encoding a proteinase, ech42 encoding an endochitinase, and a β-glucanase gene, were determined. Promoter elements in the prb1 and ech42 genes suggested that transcription is regulated by carbon and nitrogen levels and by stress. Both genes had mycoparasitism-related elements indicating potential roles for the protein products in competition. The promoter of the β-glucanase gene contained CreA and AreA binding sites indicative of catabolite regulation but contained no mycoparasitism elements. Transcription of the 3 genes was measured in mixed cultures of T. aggressivum and A. bisporus. Two A. bisporus strains, U1, which is sensitive to green mould disease, and SB65, which shows some resistance, were used in co-cultivation tests to assess possible roles of the genes in disease production and severity. prb1 and ech42 were coordinately upregulated after 5 days, whereas β-glucanase transcription was upregulated from day 0 with both Agaricus strains. Upregulation was much less pronounced in mixed cultures of T. aggressivum with the resistant strain, SB65, than with the sensitive strain, U1. These observations suggested that the proteins encoded by these genes have roles in both nutrition and in severity of green mould disease.

A new method to apply and quantify bruising sensitivity of button mushrooms

Mushrooms are prone to develop brown discolouration due to bruising caused by mechanical damage during harvest, which leads to reduced quality of the mushrooms. In order to study the mechanism behind discolouration and to breed for bruise-related browning resistant strains, a high throughput bruise application method and reliable quantification of browning sensitivity is essential. In this study a new bruising device was developed to bruise mushrooms in a fast and reproducible way. The bruising device can apply damage to the cap tissue of button mushrooms by a slip-shear sliding process. The severity of the bruise was quantified with a computer imaging system combined with a newly developed software programme. A protocol was developed to obtain the most reliable and reproducible method to compare browning sensitivity of different Agaricus bisporus strains. For this purpose, the extent of damage applied, time between harvest and bruising, developmental stage of the mushrooms and flushes were taken into account. The new method of bruising and image based quantification was subsequently applied to a collection of wild, commercial and hybrid A. bisporus strains and a distinction could be made between bruise-related browning sensitive mushrooms and tolerant mushrooms.

The extreme xerophilic mould Xeromyces bisporus — Growth and competition at various water activities

Little is known about the mould, Xeromyces bisporus, unique in its strong xerophilicity and ability to grow at water activity ( a w) 0.62, lower than for any other known organism. The linear growth rates of one fast and one slow-growing strain of X. bisporus were assessed at 20, 25, 30 and 37 °C on solid agar media containing a mixture of glucose and fructose to reduce a w to 0.94, 0.88, 0.84, 0.80, 0.76 and 0.66. Growth rates of xerophilic species closely related to X. bisporus, viz. Chrysosporium inops, C. xerophilum and Monascus eremophilus, were also assessed. Optimal conditions for growth of both X. bisporus strains were approx. 0.84 a w and 30 °C, despite FRR 2347 growing two- to five-fold faster than CBS 185.75. X. bisporus FRR 2347 even grew well at 0.66 a w (0.48 mm/day). C. inops and C. xerophilum were more tolerant of high a w than X. bisporus, and could be differentiated from each other based on: the faster growth of C. xerophilum; its preference for temperatures ≥ 30 °C and a w ≥ 0.94 ( c.f. ≤ 25 °C and ~ 0.88 a w for C. inops); and its ability to grow at 0.66 a w, which is the lowest a w reported to date for this species. M. eremophilus grew slowly (max. 0.4 mm/day) even in its optimal conditions of ~ 0.88 a w and 25 °C. To investigate the competitive characteristics of X. bisporus at low a w, both X. bisporus strains were grown in dual-culture with xerotolerant species Aspergillus flavus and Penicillium roqueforti, and xerophilic species A. penicillioides, C. inops, C. xerophilum and Eurotium chevalieri, on glucose–fructose agar plates at 0.94, 0.84, 0.80 and 0.76 a w and at 25 °C. Growth rates and types of interactions were assessed. Excretion of inhibitory substances acting over a long-range was not observed by any species; inhibitors acting over a short-range that temporarily slowed competitors' growth or produced a protective zone around the colony were occasionally observed for A. penicillioides, C. inops and C. xerophilum. Instead, rapid growth relative to the competitor was the most common means of dominance. The xerotolerant species, A. flavus and P. roqueforti were dominant over X. bisporus at 0.94 a w. E. chevalieri was often dominant due to its rapid growth over the entire a w range. At a w < 0.80, X. bisporus was competitive because it grew faster than the other species examined. This supports the concept that its ideal environmental niche is sugary foods with low a w.

Crescimento micelial in vitro de cinco linhagens de Agaricus bisporus submetidas a diferentes condições de temperatura

The in vitro mycelium growth of Agaricus bisporus strains ABI-05/03, ABI-06/04, ABI-04/02, ABI-06/05 and ABI-01/01 was evaluated in solid culture media made up of compost. Evaluations were performed by means of measurements of four diameters of the colonies, every 48 hours, during 12 days of incubation in darkness under 20 and 25oC. The experimental design consisted of randomized blocks, using the Tukey test to compare averages. Based on the obtained results, it was verified that: mycelium growth of A . bisporus is influenced by the temperature of incubation; the temperature of 25oC was more favorable to the mycelium growth of all A . bisporus strains; under the temperature of 20oC, the best growth was obtained with strains ABI-06/05 and ABI-01/01 and, under the temperature of 25oC, strain ABI-01/01 showed significantly higher growth than all other strains.

Crescimento micelial in vitro de cinco linhagens de Agaricus bisporus submetidas a diferentes condições de temperatura

The in vitro mycelium growth of Agaricus bisporus strains ABI-05/03, ABI-06/04, ABI-04/02, ABI-06/05 and ABI-01/01 was evaluated in solid culture media made up of compost. Evaluations were performed by means of measurements of four diameters of the colonies, every 48 hours, during 12 days of incubation in darkness under 20 and 25&ordm;C. The experimental design consisted of randomized blocks, using the Tukey test to compare averages. Based on the obtained results, it was verified that: mycelium growth of A. bisporus is influenced by the temperature of incubation; the temperature of 25&ordm;C was more favorable to the mycelium growth of all A. bisporus strains; under the temperature of 20&ordm;C, the best growth was obtained with strains ABI-06/05 and ABI-01/01 and, under the temperature of 25&ordm;C, strain ABI-01/01 showed significantly higher growth than all other strains.

Yield of four Agaricus bisporus strains in three compost formulations and chemical composition analyses of the mushrooms

Three compost formulations, consisting of two varieties of Cynodom dactylon (L.) Pers. (Coast-cross and Tyfton) and oat (Avena sativa) straw were tested for the cultivation of A. bisporus strains ABI-01/01, ABI-04/02, ABI-05/03, and ABI-06/04. A completely randomized experimental design in a factorial scheme was adopted, with 12 treatments (4 A. bisporus strains × 3 types of compost) and 8 replicates. Each experimental unit corresponded to one box containing 12 12.5 kg fresh wet compost. The data were submitted to analysis of variance and the means were compared by Tukey test. According to the results, productivity of mushrooms was influenced by strain and/or compost type. It was also verified that crude protein, ash, and crude fiber contents in the mushroom varied with A. bisporus strain and straw used in the formulation of the compost.

Differentiation of commercial strains of Agaricus species in China with inter-simple sequence repeat marker

In recent years, Agaricus mushroom production has undergone an important increase in the world. However, there is no consensus over the taxonomy of some commercial strains. So rapid and reliable methods to identify Agaricus strains are urgently needed. In this paper, six inter-simple sequence repeat (ISSR) primers were screened from 20 primers to analyse the genetic diversities of the 12 main commercial strains of Agaricus in China. The results showed that a total of 124 bands were produced, among which 80.6% was polymorphic, the similarity coefficients ranged from 0.69 to 0.98. Twelve tested strains were divided into 2 groups on the similarity coefficient of 0.75. Three Agaricusbitorquis strains were clustered in one group, and seven Agaricus bisporus strains were clustered with Agaricus strains S1 and S2 in the other group. As comparison with ISSR, intergenic spacers combined with restriction fragment length polymorphisms (IGS-RFLP) was employed to analyse genetic diversities of the tested strains by digesting IGS amplified products with restriction enzymes Hae III, Alu I, Rsa I, respectively, and IGS-RFLP cluster results were similar with those of ISSR. However, ISSR was superior to IGS-RFLP in differentiating closely related strains. It is suggested that the ISSR marker is an alternative method to differentiate Agaricus strains.

Yield of four Agaricus bisporus strains in three compost formulations and chemical composition analyses of the mushrooms.

Three compost formulations, consisting of two varieties of Cynodom dactylon (L.) Pers. (Coast-cross and Tyfton) and oat (Avena sativa) straw were tested for the cultivation of A. bisporus strains ABI-01/01, ABI-04/02, ABI-05/03, and ABI-06/04. A completely randomized experimental design in a factorial scheme was adopted, with 12 treatments (4 A. bisporus strains × 3 types of compost) and 8 replicates. Each experimental unit corresponded to one box containing 12 – 12.5 kg fresh wet compost. The data were submitted to analysis of variance and the means were compared by Tukey test. According to the results, productivity of mushrooms was influenced by strain and/or compost type. It was also verified that crude protein, ash, and crude fiber contents in the mushroom varied with A. bisporus strain and straw used in the formulation of the compost.

Abnormal meiosis in bisporic strains of white button mushroom Agaricus bisporus (Lange) imbach

A formerly developed method of microspreading of mushroom basidial nuclei was applied to study meiotic prophase I in bisporic white button mushroom (Agaricus bisporus) strains. Meiotic recombination and assemblage of axial structures (axial elements and synaptonemal complexes) of chromosomes in meiotic prophase I are interrelated. It is known that the frequency of meiotic recombination is reduced in the bisporic A. bisporus variety. We showed that formation of axial structures of meiotic chromosomes in bisporic strains of this mushroom was disrupted. The anomalous phenotypes in spread prophase nuclei are diverse. In leptotene and early zygotene, many nuclei contain abnormal, often short, and, as a rule, few chromosomal axial elements. The abnormalities in the formation of synaptonemal complexes at the zygotene-diplotene stage are of the same kind and even more pronounced. We discovered an important feature of meiosis in A. bisporus associated with fruit-body morphogenesis. Meiosis starting in basidia (meiocytes) of young closed fruit bodies is accompanied by disruption of chromatin condensation in prophase I and, probably, is arrested. After partial veil breakage, the course of meiosis normalizes. Preparations with clearly observable chromosomal axial structures can be obtained only at this stage of fruit-body development.

Hydrogen peroxide concentrations detected in Agaricus bisporus sporocarps and relation with their susceptibility to the pathogen Verticillium fungicola

A dry bubble is an undifferentiated structure that forms in place of mushrooms when cultures of Agaricus bisporus are contaminated by Verticillium fungicola. Hydrogen peroxide concentrations were measured in lyophilised samples of bubbles and healthy sporocarps from cultures of genetically related strains of A. bisporus. The strains the more resistant to the pathogen had the higher levels of H2O2 concentration measured in the bubbles, but the differences in the healthy sporocarps were not significant. That is an indication of a higher reaction to the pathogen in the forming sporocarps of A. bisporus strains associated with their partial resistance to V. fungicola.

Hydrogen peroxide concentrations detected in Agaricus bisporus sporocarps and relation with their susceptibility to the pathogen Verticillium fungicola

A dry bubble is an undifferentiated structure that forms in place of mushrooms when cultures of Agaricus bisporus are contaminated by Verticillium fungicola. Hydrogen peroxide concentrations were measured in lyophilised samples of bubbles and healthy sporocarps from cultures of genetically related strains of A. bisporus. The strains the more resistant to the pathogen had the higher levels of H 2O 2 concentration measured in the bubbles, but the differences in the healthy sporocarps were not significant. That is an indication of a higher reaction to the pathogen in the forming sporocarps of A. bisporus strains associated with their partial resistance to V. fungicola.

Primordia initiation of mushroom (Agaricus bisporus) strains on axenic casing materials

The mushroom (Agaricus bisporus) has a requirement for a “casing layer” that has specific physical, chemical and microbiological properties which stimulate and promote the initiation of primordia. Some of these primordia then may develop further into sporophores, involving differentiation of tissue. Wild and commercial strains of A. bisporus were cultured in axenic and nonaxenic microcosms, using a rye grain substrate covered by a range of organic and inorganic casing materials. In axenic culture, A. bisporus (commercial strain A15) was capable of producing primordia and mature sporophores on charcoal (wood and activated), anthracite coal, lignite and zeolite, but not on bark, coir, peat, rockwool, silica or vermiculite. Of six strains tested, only the developmental variant mutant, B430, produced rudimentary primordia on axenic peat-based casing material. However, none of these rudimentary primordia developed differentiated tissues or beyond 4 mm diameter, either on axenic casing material in the microcosms or in larger-scale culture. In larger-scale, nonaxenic culture, strain B430 produced severely malformed but mature sporophores in similar numbers to those of other strains. Typically, 3–6% of primordia developed into mature sporophores, but significant differences in this proportion, as well as in the numbers of primordia produced, were recorded between 12 A. bisporus strains.

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.

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 bisporus susceptibility to bacterial blotch in relation to environment: biochemical studies.

Variation of Agaricus bisporus susceptibility to bacterial blotch in relation to environment was studied at the biochemical level. Significant differences were observed between A. bisporus strains for tyrosinase and gamma-glutamyl transferase (GGT) activities and for gamma-L(+)glutaminyl-4-hydroxybenzene (GHB) content. A lower effect was detected for compost quality. GGT activity and GHB content were related to strain susceptibility to bacterial blotch by a linear model depending on strain type, commercial or wild.

Agaricus bisporus susceptibility to bacterial blotch in relation to environment: biochemical studies

Variation of Agaricus bisporus susceptibility to bacterial blotch in relation to environment was studied at the biochemical level. Significant differences were observed between A. bisporus strains for tyrosinase and γ-glutamyl transferase (GGT) activities and for γ- L(+)glutaminyl-4-hydroxybenzene (GHB) content. A lower effect was detected for compost quality. GGT activity and GHB content were related to strain susceptibility to bacterial blotch by a linear model depending on strain type, commercial or wild.

Mitochondrial Haplotype Influences Mycelial Growth of Agaricus bisporus Heterokaryons

We evaluated the influence of mitochondrial haplotype on growth of the common button mushroom Agaricus bisporus. Ten pairs of heterokaryon strains, each pair having the same nuclear genome but different mitochondrial genomes, were produced by controlled crosses among a group of homokaryons of both wild and commercial origins. Seven genetically distinct mitochondrial DNA (mtDNA) haplotypes were evaluated in different nuclear backgrounds. The growth of heterokaryon pairs differing only in their mtDNA haplotypes was compared by measuring mycelial radial growth rate on solid complete yeast medium (CYM) and compost extract medium and by measuring mycelial dry weight accumulation in liquid CYM. All A. bisporus strains were incubated at temperatures similar to those utilized in commercial production facilities (18, 22, and 26(deg)C). Statistically significant differences were detected in 8 of the 10 heterokaryon pairs evaluated for one or two of the three growth parameters measured. Some heterokaryon pairs showed differences in a single growth parameter at all three temperatures of incubation, suggesting a temperature-independent difference. Others showed differences at only a single temperature, suggesting a temperature-dependent difference. The influence of some mtDNA haplotypes on growth was dependent on the nuclear genetic background. Our results show that mtDNA haplotype can influence growth of A. bisporus heterokaryons in some nuclear backgrounds. These observations demonstrate the importance of including a number of mitochondrial genotypes and evaluating different nuclear-mitochondrial combinations of A. bisporus in strain improvement programs.

Activation of tyrosinase in Agaricus bisporus strains following infection by Pseudomonas tolaasii or treatment with a tolaasin-containing preparation

Three A. bisporus strains (U1, C45 and B62) with different resistance towards bacterial blotch were infected with Pseudomonas tolaasii or with a partly purified toxin extract. In the two susceptible strains (U1 and B62), the total tyrosinase level decreased upon treatment with bacteria or the toxin-containing extract. The amount of active tyrosinase increased, suggesting an activation of tyrosinase upon infection. In the resistant strain (C45), the tyrosinase activity remained unchanged. Protease levels were found to be increased in regions of the sporocorp that were infected with the pathogenic bacterium, but not when treated with the toxin preparation, suggesting an activation of tyrosinase by a non-proteolytic mechanism. This observation was confirmed with in vitro experiments in which tyrosinase was found to be activated by a protease-free tolaasin-containing preparation.