Volume 10, Part 2, May 1996

CULTIVATING MUSHROOMS: MAKING COMPOSTED AND NON-COMPOSTED SUBSTRATES

RICHARD SCRASE
'Mushroom makers', 38 Eastbourne Avenue, Bath BA1 6EN, UK Tel./Fax: 01255338165/01225447962
This article describes the range of materials used for mushroom cultivation and introduces the methods for their preparation. The production of sufficient mycelial biomass to support a useful mushroom yield has traditionally taken place within large quantities of naturally occurring materials. These can be relatively unprocessed, ego freshly cut wood (logs), or transformed by procedures known as 'composting' The compost (using the term loosely, as some substrates mentioned have not necessarily been through a composting process of alteration by microorganisms) not only provides the nutritional requirements of the fungus but also the physical and chemical conditions that give the fungus a competitive edge over other microorganisms. All saprotrophic basidiomycetes require carbon compounds which are usually obtained from plant polymers. The enzymes that break down these polymers can together be effective against a wide range of cellular architectures, and so have an action that is non-specific enough to allowthe grower to replace the natural diet with more readily available alternatives, ego substituting straw for wood. Nitrogen compounds also have to be provided and sources containing the nitrate ion are usually acceptable. Essential elements and vitamins are not usually growth-limiting except for calcium which is often added to regulate pH and to modify the structure of the compost. Almost any organic material can be composted, but it must be mixed to give a carbon/nitrogen ratio of between 80:1 and 10:1 (Charlesworth, 1995). Chemical analysis of wood shows a C/N ratio of 500-1250:1 (Stamets & Chilton, 1983). Other natural substrates have nitrogen contents that are growth-limiting so nitrogen supplementation increases mushroom yields in many compost systems. One example is Lepista nuda (Bull. : Fr.)Cooke, the Wood Blewit; this fungus utilises leaf-litter in nature but has been grown semi-commercially on Agaricus bisporus (Lange)Imbach mushroom compost (Frische & VanLoone, 1989). The fact that Lepista nuda can fruit on this range of composts demonstrates a fundamental ecological niche for the Wood Blewit that is wider than its perceived ('realised') niche in the wild. This contrast between the perceived and fundamental niches may well be found in other species, in which case the example of Lepista nuda is a message of hope to the cultivator because it raises the possibility that the composts listed below may serve as suitable substrates for many different species. Those fungi which are currently under largescale cultivation are grown on a spectrum of composts; from non-sterile fresh lignocellulosic material such as straw used for Pleurotus spp. (Zadrazil & Reiniger, 1988) and logs used for Lentinus edodes (Berk.)Sing. (Przybylowicz & Donoghue, 1990) through Volvariella volvacea (Bull. : Fr.)Singer (Chang & Hayes, 1978) on straw composted for a few days, to Agaricus bisporus on straw supplemented with nitrogen (manure) composted in two phases over three weeks. The processes of compost preparation will only be outlined below, fuller treatments are available in the references (Stamets & Chilton, 1993; Fermor et aI, 1985).

Natural bulk substrates

The use of non-composted materials is common among commercial growers of Pleurotus, Volvariella, Flammulina and Stropharia. These materials can be sterilised, pasteurised, or used untreated in their natural state. For example I have grown Pleurotus ostreatus (Jacq. : Fr.)Kummer outdoors on a 'bale' of unsterilised wheat straw, soaked in cold tap-water, with good results. The Pleurotus mycelium grew in all but the outer 5cm of the bale. When growing mushrooms indoors however, problems with fungal and other contaminants make it prudent to pasteurise or sterilise the compost. Steineck (1984) lists many species he has grown on straw or wood-chips buried in damp and shady places in his garden and these materials can form the basis of both garden and commercial

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scale cultivation. growth . Because alder is high in sugar content, without resins, and abundant, it has often been selected (Stamets & Chilton, 1983). In spring, fresh cut branches are chipped, mixed wit h the inoculum (which itself can be grown on wood-chips) and made into a ridge bed on the ground. Pieces 2-8 em long give better results than sawdust in non-sterile conditions. The bed size commonly used is 60 em wide by 15 em deep. Incidentally, the beds of wood-chips and bark used to cushion falling children in parks frequently support massive fruitings of mushrooms, ego Hypholoma fasciculare (Huds.: Fr.)Kummer (sulphur tuft) and the false morel Gyromitra esculenta (Pers.)Fr. (personal observation).

Cereal straw
Straw, clean and unspoiled by any previous decomposition, is prepared by chopping it into 2-8 cm lengths and soaking in water for about three days to achieve 75% moisture. Baled cereal straw is ideal as it needs no further chopping. This is sufficient for competitive species such as Pleurotus ostreatus but to reduce losses from competition the straw can be pasteurised by using hot water or sterilised by steam. Straw is usually ino culated with grain spawn at the rate of 1-2% by weight (Stamets & Chilton, 1983).

Deciduous leaves
Leaves were used by Matruchot (1917) to grow Lepista nuda in caves left by limestone mining. In the process of replicating his experiments in a local disused mine , I discovered that standard mushroom compost could be used as a substrate instead. The yields from mushroom compost are higher, but it is generally used in a less romantic setting . Leaf mould has a long history of use in horticulture , traditionally by being stacked outdoors in piles 60-90 em deep for t wo years to allow it to decompose (Larkcom, 1976). The resulting fibrous material is used as a mulch or as the water-holding component of seed, cutting and potting composts. The need to replace peat (Pryce, 1991) may create a niche for leaf-mould. Preliminary experiments using leaves inoculated with Lepista nuda produced horticulturally useful leaf-mould in six months (Scrase, 1993). Leaves are often available in the autumn from civic authorities and should not be collected from the wild.

Sawdust
The particulate nature of sawdust makes nutrients more readily available, consequently competing organisms can rapidly invade these materials. Therefore the sawdust-based substrate is usually sterilised or pasteurised prior to inoculation. Generally the sawdust type is matched to the fungus, but alternatives to the usual host species can be substituted. Even softwood sawdust can be substituted for hardwood if it is aged for about a year to allow the reduction in the amount of resins and phenolics ; more commonly growers use a blend. The time taken to degrade the substratum is related to particle size and with shiitake the optimum is 2-3 mm (Nisikado et al, 1942). Smaller particles reduce gas-flow and restrict growth. Sawdust is usually supplemented to increase the amount of nitrogen and easily assimilated carbohydrates. While high levels of supplementation result in higher yields it also encourages the growth of competitors . The recipes below are a result of finding a balance between these two opposing considerations (Stamets & Chilton, 1983). standard formula sawdust 80% cereal bran 20% low contamination formula sawdust 90% cereal bran 10%

Wood-logs
The most notable commercial species grown on wood is Lentinus edodes or Shiitake. Logs (traditionally oak) 7-15 em in diameter and 90 em long are cut between autumn and spring when sap content is highest. Care is taken to leave the bark intact as this has consequences for subsequent fruiting. Holes 2-3 cm deep are drilled 20 em apart into the log, then plugged with sawdust spawn or mycelium-colonised dowels (Przybylowicz & Donoghue, 1990).

Wood-chips
Freshly cut chips of alder (Alnus glutinosus), maple (Acer spp.), and fir (Abies spp.) can support mycelial

The optimum moisture level is between 55% and 70% by dry weight before heat treatment. This can be established by oven-drying a sample ; however many growers estimate the proper moisture content by squeezing the mixture in one hand ; drops of free water should be squeezed out. Air exchange is enabled by using low density polyethylene bags or by inserting cotton wool, foam,

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etc. into the neck of the container holding the substrate. inorganic nitrogen can be used. Elephant manure has been found satisfactory (Genders , 1969). The aim is to create an initial mixture with a nitrogen content of 1.5-1.7%, a water content of 70% (+/-3%), and a carbon/nitrogen ratio of approximately 30:1. Many materials can be used as supplements to modify the C/N ratio, egourea, cereal bran, or sugar beet pulp. The following recipes are based on using fresh materials but are given as dry weight ratios . Horse manure has proportions of manure and straw 100 to gypsum 5 while synthetic compost is made up in these proportions: wheat straw 52, chicken manure 45, gypsum 3.5. To make mushroom compost, fresh manure and straw is mixed, soaked and then stacked. To prevent the development of anaerobic conditions within the stack the vertical cross-section is generally not more that 2 x 2 metres. This allows the microorganisms present in the raw materials to begin to degrade the stack. Their activity heats up the stack which in turn makes conditions ideal for further decomposition by other microorganisms. The stack core is depleted of oxygen in 48 to 96 hours and it is then rebuilt. This procedure is repeated three times during the first phase to allow decomposition to continue, typically during the third, fifth and seventh day after initial mixing. The centre of the stack, should reach 65-80C. At the end of this first phase the compost should be deep brown, be flecked with whitish colonies of actinomycetes , smell of ammonia, have a pH of 8.08.5, and release liquid when firmly squeezed. In the second phase, the compost is filled into trays , beds or bags and pasteurised (58-60C). This kills nematodes, fly eggs and larvae, mites and competitor fungal mycelium and spores. The compost is 'fermented' for a further 5-8 days. During this time the internal temperature of the compost is held at about 50C by controlling ventilation. This is optimal for the actinomycetes that incorporate ammonia at this stage. The presence or rather absence of ammonia is usually detected by smell, but cresyl orange indicator or gas detection tubes can be used. After the ammonia concentration has dropped to 10-20 [lgg-l, the temperature is allowed to drop to 25C and the compost is inoculated. In the process of 'long composting' the whole composting procedure takes place outdoors . The stack is turned four times, on days 6, 10-12, 13-15 and 15-17. The anaerobic centre zone is moved to the outside on each occasion. The finished compost is pasteurised for four hours. If this is not pos-

Other lignocellulosic wastes A tremendous range of materials has been used, especially for the growth of Pleurotus spp. ego sugar cane bagasse, pomace (apple waste), sugar beet pulp, coffee, tea, cotton wastes etc. Unless the waste contains a toxic component, it is likely that by adjusting moisture levels to around 70%, providing aeration, and optimising carbon/nitrogen ratios, any plant waste can be degraded by basidiomycete fungi. At the 1995 congress on the science and cultivation of edible fungi delegates reported on several examples of using basidiomycetes to transform waste materials , egoAuricularia polytricha has been grown on coffee wood in Mexico, and Pleurotus ostreatus used to turn waste-paper into horticultural mulching material in the Seychelles.

Agaricus bisporus compost

Fermor et al (1985) give a very full account of the processes involved in preparing compost for Agaricus bisporus while Genders (1969) gives a more colloquialdescription; the following howeveris drawn primarily from Stamets & Chilton (1983). I personally obtain mushroom compost from a commercial compost producer, complete except for the inoculation of mushroom spawn. The commercial composting process is divided into two stages, commonly called Phase 1 and Phase 2, but the process, called 'Long composting' is more appropriate for the amateur grower, as there is no need for the environmentally controlled rooms used for phase 2 processing. Both systems willbe described here. The raw materials for the compost are straw, manure, calcium sulphate (gypsum), water and nutritional supplements. Wheat straw is preferred because its resilience maintains an open well-ventilated compost structure. The straw is the primary source of carbohydrates and in addition it provides lignin that is converted into the nitrogen rich ligninhumus-complex, which provides a source of protein to the mushroom mycelium. Calcium sulphate improves compost structure by aggregating colloidal particles and it also provides a source of metabolic calcium. Horse-manure (usually ready mixed with bedding-straw) is the traditional source of nitrogen; while so called 'synthetic' composts are made with chicken manure, but other sources of organic and

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sible, discard the outer shell of the stack, and use the compost showing the white flecking indicating strong actinomycete activity. Inoculation and spawn running Mushroom spawn is mixed into the compost, usuallyas 1-2% by wet weight. Conditions then need to be maintained for the strain in question. The primary requirement is to maintain the core compost temperature at the optimum temperature. This is 25C for many commercial strains of Agaricus, Lepiota, Lepista and Coprinus. The compost usually contains sufficient oxygen and water to support the development of the fungal mycelium for the duration of the growth period. The success of themycelial growth or 'spawn-run' can be seen by the change in colour of the compost. The whitish colour of mycelium is also evident. Similar changes can also be seen with other compost types, leafmould and straw become paler as the fungal enzymes bleach the substrate. However, the apparanently successful utilisation of the substrate is no guarantee of success with mushroom production. On several occasions I have had straw bales smelling sweetly of fungal decomposition, all but the outermost layer of straw charged with mycelium, ready to produce ..nothing! In essence the mycelium is induced to go through a developmental switch from mycelial extension to the production of mushroom primordia. Conditions are then maintained that facilitate the development of these primordia into mushrooms. The initial switch is usually achieved by radically altering the environmental conditions surrounding the mycelium; typically by reducing temperature and increasing oxygen supply. Subsquent development then requires higher humidity (Flegg & Wood, 1985). The understanding of the mechanisms of basidiomycete morphogenesis have yet to be synthesised into a complete story; some of the components can be found in Moore et al (1985), but this area is undoubtedly a major problem in the scientific understanding of mushroom growing. We cannot yet definitely say why on some occasions mushrooms fruit from a mycelium, and on other occasions they do not, it is another part of life's rich complexity. The methods for initiating and maintaining fruiting willform the basis for a further article.

References
Charlesworth, K. (1995) Life, the universe & (almost) everything. Muck and Magic.New Scientist 1979:53. Chang, S.T. & Hayes, W.A. (eds) (1978) The Biology and Cultivation of Edible Mushrooms. Academic Press, New York. Fermor, TR, Randle, P.E. & Smith, J.F. (1985) Compost as a substrate and its preparation. In P.E. Flegg, D.M. Spencer, D.A. Wood (eds): The Biology and Technology of the Cultivated Mushroom. John Wiley & Sons: Chichester & New York. pp. 81-110. Flegg, P.E. & Wood, D.A. (1985) Growth and fruiting. In P.E. Flegg, D.M. Spencer, D.A. Wood (eds): The Biology and Technology of the Cultivated Mushroom. John Wiley & Sons: Chichester & New York. pp. 141-178. Frische, G. & vanLoone, P. (1989) Breeding experiments with the wood blewit (Lepista nuda). In Mushroom Science XII (partl), Proceedings of the Twelfth

International Congress on the Science and Cultivation of Edible Fungi, Germany (1987). Genders, R. (1969) Mushroom growing for everyone. Faber &
Faber, London. Larkcom, J. (1976) Vegetables from small gardens. Faber & Faber, London. Matruchot, L. (1917) Variations experimentales due Tricholoma nudum. Revue Generale Botanique. 503. Moore, D., Casselton, L.A., Wood, D.A. & Frankland, J.C. (eds) (1985) Developmental Biology of Higher Fungi. BMS Symposium 10, Cambridge University Press. Nisikado, Y., Kimura, K. & Miyawaki, T. (1942) The mycelial growth in pure culture on the sawdust medium prepared of various kinds of tree. Ber. Ohara Inst.

Landw. Forsch. 9:39-60.

Pryce, S. (1991) The Peat Alternatives Manual. Friends of the Earth, London. Przybylowicz, P. & Donoghue, J. (1990) Shiitake Growers Handbook, Kendall/Hunt, Iowa. Scrase, R.J. (1993) The Cultivation of Basidiomycetes. M Phil thesis, Bath University. Stamets, P. & Chilton, J.S. (1983). The Mushroom Cultivator, Agarikon Press, Olympia, Washington. Steineck, H. (1984) Mushrooms in the Garden. Mad River Press, California. Zadrazil, F. & Reiniger, P. (eds) (1988) Treatment of lignocellulosics with white rot fungi. Elsevier, London.