Physicochemical characterisation of casings in relation to mushroom (Agaricus bisporus) cropping performance.

Peat-based casings have been used for button mushroom (Agaricus bisporus) cultivation for decades but there is environmental pressure to find sustainable alternatives. This work aimed to characterise the physicochemical properties of peat and peat-substituted casings and to determine their influence on mushroom cropping to enable alternatives to be identified. British milled peat and German wet-dug peat casings produced smaller mushrooms than Irish wet-dug peat casing although yield was unaffected. Substitution of milled or wet-dug peat casings with 25% v/v bark, green waste compost or spent mushroom casing, except Irish wet-dug peat casing with spent peat mushroom casing, caused reductions in mushroom yield and/or size. These poorer results of casings compared with Irish wet-dug peat casing corresponded with lower water retention volumes at matric potential (Ψm) -15 kPa but not after drainage from saturation or at -1 kPa. Air-filled porosity (17-22% v/v), compacted bulk density after drainage (670-800 g L-1) and electrical conductivity (0.30-0.54 mS cm-1) of casings were unrelated to their mushroom cropping performance. In-situ casing measurements with electronic tensiometers confirmed laboratory casing physical analysis: at the same casing Ψm, Irish wet-dug peat casing had a higher water content than German wet-dug peat casing and produced larger mushrooms for the same yield. Solid-state foam-based tensiometers were more robust than water-filled tensiometers but they did not detect the full decrease in casing Ψm during a flush of mushrooms. The results indicate that if sustainable materials are to replace wet-dug peat casing with the same mushroom yield and size quality performance, they should have equivalent water retention volumes at Ψm -15 kPa. Measurement of casing Ψm with electronic tensiometers to control mushroom crop irrigation should assist in this transition.

Nitrogen balance and supply in Australasian mushroom composts.

Mushrooms are an important source of protein in the human diet. They are increasingly viewed as a sustainable meat replacement in an era of growing populations, with button mushrooms (Agaricus bisporus) the most popular and economically important mushroom in Europe, Australia and North America. Button mushrooms are cultivated on a defined, straw-derived compost, and the nitrogen (N) required to grow these high-protein foods is provided mainly by the addition of poultry manure and horse manure. Using the correct balance of carbon (C) and N sources to produce mushroom compost is critically important in achieving maximum mushroom yields. Changes in the amount and form of N added, the rate and timing of N addition and the other compost components used can dramatically change the proportion of added N recovered in the mushroom caps, the yield and quality of the mushrooms and the loss of N as ammonia and nitrogen oxide gases during composting. This review examines how N supply for mushroom production can be optimised by the use of a broad range of inorganic and organic N sources for mushroom composting, together with the use of recycled compost leachate, gypsum and protein-rich supplements. Integrating this knowledge into our current molecular understanding of mushroom compost biology will provide a pathway for the development of sustainable solutions in mushroom production that will contribute strongly to the circular economy. KEY POINTS: • Nitrogen for production of mushroom compost can be provided as a much wider range of organic feedstocks or inorganic compounds than currently used • Most of the nitrogen used in production of mushroom compost is not recovered as protein in the mushroom crop • The sustainability of mushroom cropping would be increased through alternative nitrogen management during composting and cropping.

Molecular characterization of Pseudomonas from Agaricus bisporus caps reveal novel blotch pathogens in Western Europe.

BACKGROUND: Bacterial blotch is a group of economically important diseases affecting the cultivation of common button mushroom, Agaricus bisporus. Despite being studied for more than a century, the identity and nomenclature of blotch-causing Pseudomonas species is still unclear. This study aims to molecularly characterize the phylogenetic and phenotypic diversity of blotch pathogens in Western Europe. METHODS: In this study, blotched mushrooms were sampled from farms across the Netherlands, United Kingdom and Belgium. Bacteria were isolated from symptomatic cap tissue and tested in pathogenicity assays on fresh caps and in pots. Whole genome sequences of pathogenic and non-pathogenic isolates were used to establish phylogeny via multi-locus sequence alignment (MLSA), average nucleotide identity (ANI) and in-silico DNA:DNA hybridization (DDH) analyses. RESULTS: The known pathogens "Pseudomonas gingeri", P. tolaasii, "P. reactans" and P. costantinii were recovered from blotched mushroom caps. Seven novel pathogens were also identified, namely, P. yamanorum, P. edaphica, P. salomonii and strains that clustered with Pseudomonas sp. NC02 in one genomic species, and three non-pseudomonads, i.e. Serratia liquefaciens, S. proteamaculans and a Pantoea sp. Insights on the pathogenicity and symptom severity of these blotch pathogens were also generated. CONCLUSION: A detailed overview of genetic and regional diversity and the virulence of blotch pathogens in Western Europe, was obtained via the phylogenetic and phenotypic analyses. This information has implications in the study of symptomatic disease expression, development of diagnostic tools and design of localized strategies for disease management.

Environmental regulation of reproductive phase change in Agaricus bisporus by 1-octen-3-ol, temperature and CO2.

Reproductive phase change from vegetative mycelium to the initiation of fruiting in Agaricus bisporus is regulated in large part by the sensing of environmental conditions. A model is proposed in which three separate environmental factors exert control at different stages of the reproductive developmental process change. The eight carbon volatile 1-octen-3-ol controls the early differentiation from vegetative hyphae to multicellular knots; temperature reduction is essential for the later differentiation of primodia; and carbon dioxide level exerts quantitative control on the number of fruiting bodies developed. Analysis of transcriptomic changes during the reproductive phase change was carried out with initiation-specific microarrays, and the newly published A. bisporus genome was used to analyse the promoter regions of differentially regulated genes. Our studies have shown there to be both early and late initiation responses relating to sensing of eight carbon volatiles and temperature respectively. A subset of 45 genes was transcriptionally regulated during the reproductive phase change which exhibited a range of functions including cell structure, nitrogen and carbon metabolism, and sensing and signalling. Three gene clusters linking increased transcription with developmental stage were identified. Analysis of promoter regions revealed cluster-specific conserved motifs indicative of co-ordinated regulation of transcription.

Volatile C8 compounds and pseudomonads influence primordium formation of Agaricus bisporus.

Primordium formation of Agaricus bisporus depends on the presence of a casing layer containing stimulatory bacteria and on sufficient air exchange. The influence of specific pseudomonad populations and volatile organic compounds (VOC) on primordium formation of A. bisporus was studied in microcosm cultures. VOC produced by A. bisporus mycelium were predominantly C8 compounds, some of which could inhibit primordium formation, with 1-octen-3-ol being most inhibitory. A VOC produced by the rye grain substrate, 2-ethyl-1-hexanol, on which A. bisporus was grown also inhibited primordium formation. 2-Ethyl-1-hexanol and 1-octen-3-ol were metabolized by pseudomonad populations and adsorbed by activated charcoal, with both modes of removal enabling primordium formation in the casing. Removal of VOC by ventilation also enabled primordium formation to occur under axenic conditions. The presence of 2-ethyl-1-hexanol and 1-octen-3-ol in the microcosms resulted in higher total bacterial and pseudomonad populations in the casing. The stimulatory effects of the casing and its microbiota and air exchange on primordium formation of A. bisporus at least partly are due to the removal of inhibitory C8 compounds produced by the mycelium and its substrate. Monitoring and controlling the levels of these inhibitory VOC in mushroom culture should enable primordium formation of A. bisporus to be more efficiently and precisely controlled.

Aerating recycled water on mushroom composting sites affects its chemical analysis and the characteristics of odor emissions.

Recycled water (RW) containing compost leachate can be a potent source of foul odor on mushroom composting sites. Samples of RW were repeatedly collected from storage tanks and pits of 14 mushroom composting sites in England and Ireland. Relationships between the effects of submerged aeration of the RW, the chemical and microbial characteristics of the RW, and the odors emitted were investigated. Recycled water samples were analyzed for pH, electrical conductivity (EC), redox potential, and dissolved oxygen concentration after 7 to 14 d cold storage. Freeze-dried material from the RW samples was chemically profiled by pyrolysis gas chromatography-mass spectrometry (GC-MS), and the content of odor precursor compounds was determined by the ninhydrin colorimetric method. The headspace air of containerized RW samples was analyzed by thermal desorption GC-MS and with gas detection tubes and assessed for odor intensity (OI) and concentration by panelists and serial dilution olfactometry. The predominant odorants in the headspace and freeze-dried residues of RW samples were volatile sulfur-containing compounds and carboxylic acids. The headspace OI, EC, dry matter content, and redox potential of RW were interrelated. The headspace OI and combined concentration of hydrogen sulfide (H(2)S) + dimethyl sulfide of RW were correlated (R(2) = 0.635; P < 0.001). Prediction of the OI of RW by measuring RW EC and the concentration of headspace sulfides using gas detector tubes enables rapid and low-cost monitoring of RW from mushroom composting sites. Submerged aeration of RW in storage tanks or pits reduced the RW headspace air OI and the combined H(2)S + dimethyl sulfide concentration by 88%.

Characterization of recycled mushroom compost leachate by chemical analysis and thermogravimetry-mass spectrometry.

Recycled compost leachate (RCL or euphemistically named "goody water") can be a potent source of foul odor on mushroom substrate production sites and contributes to composting smells. A complex mixture of sulfur compounds, fatty acids, and nitrogen containing compounds is responsible for odor production. Fifty samples, collected from 14 compost production sites in Ireland and the U.K. over a 2 year period, were analyzed for chemical properties and by thermogravimetry-mass spectrometry (TG-MS) for compositional differences. Results indicated that aerated samples had lower values of electrical conductivity, redox potential, and dry matter content than nonaerated samples and that the higher thermal stability of aerated samples measured by TGA could be attributed to greater mineralization of the substrate due to aerobic processes. The lower temperatures noted for peak evolution of methane, water, and carbon dioxide from TG-MS analysis suggested that a more energetic process had occurred in aerated RCL storage facilities, producing greater decomposition of macromolecules that volatilized at lower temperatures. Chemical composition, thermal stability of the freeze-dried leachate, pyrolysis profiles, and relative amounts of pyrolysis products were all markers of as to how effective control measures could influence RCL quality.