The significance of volatile antifungal metabolites produced by trichomerma harzianum biotype Th4, in green-mould disease of commercial mushroom crops /

The aggressive mushroom competitor, Trichoderma harzianum biotype Th4, produces volatile antifungal secondary metabolites both in culture and during the disease cycle in compost. Th4 cultures produced one such compound only when cultured in the presence of Agaricus bisporus mycelium or liquid medium made from compost colonised with A. bisporus. This compound has TLC and UVabsorption and characteristics indicating that it belongs to a class of pyrone antibiotics characterised from other T. harzianum biotypes. UV absorption spectra indicated this compound was not 6-pentyl-2H-pyran-one (6PAP), the volatile antifungal metabolite widely described in Th1. Furthermore, this compound was not produced by Th1 under any culture conditions. Mycelial growth of A. bisporus, Botrytis cinerea and Sclerotium cepivorum was inhibited in the presence of this compound through volatility , diffusion and direct application. This indicates that Th4 produces novel, volatile, antifungal metabolites in the presence of A. bisporus that are likely involved in green mould disease of mushroom crops.

Cell wall degrading enzymes and interaction between Trichoderma Aggressivum and Agaricus Bisporus

Agaricus bisporus is the most commonly cultivated mushroom in North America and has a great economic value. Green mould is a serious disease of A. bisporus and causes major reductions in mushroom crop production. The causative agent of green mould disease in North America was identified as Trichoderma aggressivum f. aggressivum. Variations in the disease resistance have been shown in the different commercial mushroom strains. The purpose of this study is to continue investigations of the interactions between T. aggressivum and A. bisporus during the development of green mould disease. The main focus of the research was to study the roles of cell wall degrading enzymes in green mould disease resistance and pathogenesis. First, we tried to isolate and sequence the N-acetylglucosaminidase from A. bisporus to understand the defensive mechanism of mushroom against the disease. However, the lack of genomic and proteomic information of A. bisporus limited our efforts. Next, T. aggressivum cell wall degrading enzymes that are thought to attack Agaricus and mediate the disease development were examined. The three cell wall degrading enzymes genes, encoding endochitinase (ech42), glucanase (fJ-1,3 glucanase) and protease (prb 1), were isolated and sequenced from T. aggressivum f. aggressivum. The sequence data showed significant homology with the corresponding genes from other fungi including Trichoderma species. The transcription levels of the three T. aggressivum cell wall degrading enzymes were studied during the in vitro co-cultivation with A. bisporus using R T -qPCR. The transcription levels of the three genes were significantly upregulated compared to the solitary culture levels but were upregulated to a lesser extent in co-cultivation with a resistant strain of A. bisporus than with a sensitive strain. An Agrobacterium tumefaciens transformation system was developed for T. aggressivum and was used to transform three silencing plasmids to construct three new T. aggressivum phenotypes, each with a silenced cell wall degrading enzyme. The silencing efficiency was determined by RT-qPCR during the individual in vitro cocultivation of each of the new phenotypes with A. bisporus. The results showed that the expression of the three enzymes was significantly decreased during the in vitro cocultivation when compared with the wild type. The phenotypes were co-cultivated with A. bisporus on compost with monitoring the green mould disease progression. The data indicated that prbi and ech42 genes is more important in disease progression than the p- 1,3 glucanase gene. Finally, the present study emphasises the role of the three cell wall degrading enzymes in green mould disease infection and may provide a promising tool for disease management.

Bdellovibrio bacteriovorus HD100 guards against Pseudomonas tolaasii brown-blotch lesions on the surface of post-harvest Agaricus bisporus supermarket mushrooms

BACKGROUND: Pseudomonas tolaasii is a problematic pathogen of cultured mushrooms, forming dark brown 'blotches' on mushroom surfaces and causing spoilage during crop growth and post-harvest . Treating P. tolaasii infection is difficult, as other, commensal bacterial species such as Pseudomonas putida are necessary for mushroom growth, so treatments must be relatively specific. RESULTS: We have found that P. tolaasii is susceptible to predation in vitro by the δ-proteobacterium Bdellovibrio bacteriovorus. This effect also occurred in funga, where B. bacteriovorus was administered to post-harvest mushroom caps before and after administration of the P. tolaasii pathogen. A significant, visible improvement in blotch appearance, after incubation, was observed on administration of Bdellovibrio. A significant reduction in viable P. tolaasii cell numbers, recovered from the mushroom tissue, was detected. This was accompanied by a more marked reduction in blotch severity on Bdellovibrio administration. We found that there was in some cases an accompanying overgrowth of presumed-commensal, non-Pseudomonas bacteria on post-harvest mushroom caps after Bdellovibrio-treatment. These bacteria were identified (by 16SrRNA gene sequencing) as Enterobacter species, which were seemingly resistant to predation. We visualised predatory interactions occuring between B. bacteriovorus and P. tolaasii on the post-harvest mushroom cap surface by Scanning Electron Microscopy, seeing predatory invasion of P. tolaasii by B. bacteriovorus in funga. This anti-P. tolaasii effect worked well in post-harvest supermarket mushrooms, thus Bdellovibrio was not affected by any pre-treatment of mushrooms for commercial/consumer purposes. CONCLUSIONS: The soil-dwelling B. bacteriovorus HD100 preys upon and kills P. tolaasii, on mushroom surfaces, and could therefore be applied to prevent spoilage in post-harvest situations where mushrooms are stored and packaged for sale.

Effect of supplementing compost with grapeseed meal on Agaricus bisporus production

BACKGROUND: This work assesses the agronomic performance of grapeseed meal, before and after oil extraction, in nutritional compost supplement when growing the mushroom species Agaricus bisporus (Lange) Imbach. The effect of formaldehyde treatment before using this compost is also considered. Materials were applied at different doses at spawning. Along with non-supplemented compost, three commercial nutritional supplements were used as controls.RESULTS: In general terms, grapeseed meal performance was similar to that of commercial delayed-release nutrients, but improved the non-supplemented compost response. We highlight that grapeseed enhances performance as larger yields of harvested mushrooms were obtained with greater dry weight content; however, their protein content was lower. The best performance was displayed by fresh formaldehyde-treated grapeseed (6000 ppm) when applied to the 10 g kg(-1) compost dose.CONCLUSIONS: Our findings suggest that grapeseed meal offers a great potential to be applied on a commercial scale. The addition of grapeseed resulted in an enhanced performance as shown by the higher number of harvested mushrooms. The use of grapeseed meal (extracted or non-extracted), a low-cost ingredient with high levels of carbohydrates, may suppose an economic profit on the basis of the positive effect of adding carbon in the mushroom cultivation. (C) 2012 Society of Chemical Industry

Modelling the effect of the physical and chemical characteristics of the materials used as casing layers on the production parameters of Agaricus bisporus

The aim of this research was to show the mathematical data obtained through the correlations found between the physical and chemical characteristics of casing layers and the final mushrooms' properties. For this purpose, 8 casing layers were used: soil, soil + peat moss, soil + black peat, soil + composted pine bark, soil + coconut fibre pith, soil + wood fibre, soil + composted vine shoots and, finally, the casing of La Rioja subjected to the ruffling practice. The conclusion that interplays in the fructification process with only the physical and chemical characteristics of casing are complicated was drawn. The mathematical data obtained in earliness could be explained in non-ruffled cultivation. The variability observed for the mushroom weight and the mushroom diameter variables could be explained in both ruffled and non-ruffled cultivations. Finally, the properties of the final quality of mushrooms were established by regression analysis.

A Study of Compost Added to a Casing Technique in Agaricus bisporus Cultivation from Phase III Bulk Compost

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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

Avaliou-se o crescimento micelial in vitro das linhagens ABI-05/03, ABI-06/04, ABI-04/02, ABI-06/05 e ABI-01/01 de Agaricus bisporus em meios de cultura sólidos à base de composto. As avaliações foram realizadas por meio de medições de quatro diâmetros das colônias, a cada 48h, durante 12 dias de incubação, no escuro, a 20 e 25ºC. O delineamento experimental foi o de blocos casualizados, com uso do teste de Tukey para a comparação das médias. Co m base nos resultados obtidos, verificou-se que: o crescimento micelial de A. bisporus é influenciado pela temperatura de incubação; a temperatura de 25ºC foi mais favorável para o crescimento micelial de todas as linhagens de A. bisporus; na temperatura de 20ºC, o melhor crescimento foi obtido com as linhagens ABI-06/05 e ABI-01/01; na temperatura de 25ºC, a linhagem ABI-01/01 apresentou crescimento significativamente maior que todas as demais.

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

Três formulações de composto, à base de palhas de Cynodom dactylon (L.) Pers. (cultivares Coast-cross e Tyfton) e Aveia-Avena sativa, foram testadas no cultivo das linhagens ABI-01/01, ABI-04/02, ABI-05/03 e ABI-06/04 de A. bisporus. O delineamento experimental foi em esquema fatorial, inteiramente casualizado com 12 tratamentos (4 linhagens de A. bisporus x 3 tipos de composto) e 8 repetições. Cada unidade experimental constou de uma caixa com 1212,5 kg de composto fresco úmido. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey. de acordo com os resultados obtidos verificou-se que a produção de cogumelos foi influenciada pela linhagem e/ou pelo tipo de composto. Também verificou-se que o teor de proteína bruta, cinzas e fibra bruta de basidiomas variou com a linhagem de A. bisporus e com o tipo de palha utilizada na formulação do composto.

Characterization, feasibility and optimization of Agaricus subrufescens growth based on chemical elements on casing layer

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mushroom-derived preparations in the prevention of H2O2-induced oxidative damage to cellular DNA

Aqueous extracts of the sporophores of eight mushroom species were assessed for their ability to prevent H2O2-induced oxidative damage to cellular DNA using the single-cell gel electrophoresis (Comet) assay. The highest genoprotective effects were obtained with cold (20°C) and hot (100°C) water extracts of Agaricus bisporus and Ganoderma lucidum fruit bodies, respectively. No protective effects were observed with Mushroom Derived Preparations (MDPs) from Flammulina velutipes, Auricularia auricula, Hypsizygus marmoreus, Lentinula edodes, Pleurotus sajor-caju, and Volvariella volvacea. These findings indicate that some edible mushrooms represent a valuable source of biologically active compounds with potential for protecting cellular DNA from oxidative damage. © 2002 Wiley-Liss, Inc.

Pasteuriza̧ão de turfa brasileira para o cultivo de Agaricus brasiliensis

Agaricus brasiliensis is a Brazilian basidiomycete which has been cultivated and consumed around the world as a therapeutic food. Casing layer is one of the most important steps on A. brasiliensis cultivation and European peat is the most used casing layer on Agaricus bisporus cultivation. Besides the importance of it on mushroom cultivation the peat import could be a risk of exotic organism introduction. Alternative as Brazilian peat is barely used for mushroom growers in Brazil. Thus, the objective of this work was to evaluate Brazilian peat with and without pasteurization as casing layer on A. brasiliensis cultivation. The fungus was previously grown on wheat grains and transferred to a substratum prepared by composted traditional method. After mycelium colonization of the substratum a pasteurized or non pasteurized Brazilian peat (casing layer) was added. It was concluded that pasteurization of the casing layer increases in 30% yield after 65 days of cultivation. There is no difference of yield for pasteurized and non pasteurized casing layer until 30 days of cultivation. An increase of flies is observed in non pasteurized casing layer. The production flush is easily perceived with pasteurized casing layer but not with non pasteurized casing layer.

Cultivation of Pleurotus ostreatus using supplemented spent oyster Mushroom substrate

Spent Agaricus and Pleurotus substrates are mainly used as components of amendments and growing substrates, but not in sufficient quantities to solve the problem of their accumulation in mushroom producing areas, where they represent a potential pollution risk. The mushroom growing sector in Spain generates about 5105 t of spent compost, while the EU, as a whole, produces more than 3.5106 t. Among alternative management applications, it is possible to reuse these wastes in the cultivation of mushrooms, as a casing material for growing Agaricus spp. and as substrate for growing other species. In this work, the application of commercial nutritional supplements (Calprozime, Champfood and Promycel), widely used in Agaricus cultivation, is evaluated for its possible use as additive to substrates, based on spent oyster mushroom substrate (SMS), for the cultivation of Pleurotus ostreatus. Using a mixture of straw and SMS (1:1, w/w) as base material, the addition of CaSO4 (50 g kg-1) and CaCO3 (10 g kg-1) and the above supplements at 20 g kg-1 brought about a remarkable increase in production compared with the substrate without any supplement. The biological efficiencies did not differ significantly from that obtained when a commercial substrate was used as control, reaching values of 48.9 kg/100 kg substrate (dry matter) when Calprozime was used as supplement. Sporophores harvested from the supplemented substrates presented a higher dry matter content than those obtained from both commercial and non-supplemented substrates. SMS is cheap and easily available; it can be integrated into new formulations with the added advantages of lowering production costs, limiting growers' dependence on straw, and decreasing the environmental impact of its ever-growing accumulation.

Efeito do uso do cogumelo Agaricus brasiliensis no estado nutricional, na frequência e intensidade dos efeitos adversos da terapia medicamentosa e na resposta bioquímica hepática em indivíduos com hepatite crônica pelo vírus C: estudo prospectivo, randomizado, duplo cego, placebo controlado

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

O papel de receptores de imunidade inata βGR, MR, TLR2 e TLR4 no reconhecimento da Candida albicans por monócitos humanos estimulados com polissacarídeos extraídos do cogumelo Agaricus brasiliensis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of Cultivation Practices on the beta-Glucan Content of Agaricus subrufescens Basidiocarps

Processo FAPESP: 12/15101-4