16 OVERVIEW OF MUSHROOM CULTIVATION AND UTILIZATION AS FUNCTIONAL FOODS as examples:sawdust 78%,rice/wheat bran 16%,sugar 1.5%,corn flour 1.7%. ammonium sulfate 0.3%.calcium superphosphate 0.5% and gypsum 2%;saw and gyps me 1%;and The i edodes mushroom is produced on both a cottage and a commercial seale Some issues associated with the different cultivation styles are summarized below: 1.Cottage-Scale cultivation The the substrate.The ingredients can be variable from place to place and country to country depending upon the raw materials available and local climatic conditions. In general the dry ingredi by hand or with a me hanical mixer din the The ingredients are then packed into autoclavable polypr opylene or high-density polyethylene bags.Although they are more expensive.polypropylene bags are th provides gre he closed eithe d with the preparation of the substrate for the cultivation of the mushroom are given her re as reference.(i)Sawdust 82%, wheat bran 16%. ha c 0. lime dus m14% spent c syps nhate 16 and ov sum 1 fiv Sawdust 7%wheat bran 18%com powder2%gypsum2sugar 1.%calcium superphosphate 0.5%.and urea 0.3% d sawdust medium to grov e m room.The t steps are 1)mix th (in 90 da to achieve full colonization of the sawdust mixture.in other words.to allow the mycelium to be established for ready fructification:(vi)fruit the coo sawdust logs/bags/b using a 21 dry.cut step rket The major use for humidification.and shelves to incubate Incubation can be done in two rooms and in two shipping containers.The two shipping containers fruitingrooms.The temperature dur ing incubation is etween 18 and that the blocks/los unit.With con ntalization.blocks in each room can be subjected to a cycle of humid cold,humid heat,and dry heat
16 OVERVIEW OF MUSHROOM CULTIVATION AND UTILIZATION AS FUNCTIONAL FOODS as examples: sawdust 78%, rice/wheat bran 16%, sugar 1.5%, corn flour 1.7%, ammonium sulfate 0.3%, calcium superphosphate 0.5%, and gypsum 2%; sawdust 64%, wheat bran 15%, spent coffee grounds 20%, and gypsum/lime 1%; and sawdust 78%, sucrose 1%, wheat bran 20%, and calcium carbonate 1%. The L. edodes mushroom is produced on both a cottage and a commercial scale. Some issues associated with the different cultivation styles are summarized below: 1. Cottage-Scale Cultivation There are many formulas for the composition of the substrate. The ingredients can be variable from place to place and country to country depending upon the raw materials available and local climatic conditions. In general, after mixing the dry ingredients by hand or with a mechanical mixer, water is added to the mixture so that the final moisture content of the substrate is between 55 and 60%, depending on the capacity of the sawdust to absorb water. The ingredients are then packed into autoclavable polypropylene or high-density polyethylene bags. Although they are more expensive, polypropylene bags are the most popular since polypropylene provides greater clarity than polyethylene. After the bags have been filled (1.5–4 kg wet weight) with the substrate, the end of the bag can be closed either by strings or plugged with a cottonwool stopper. Four formulas in the preparation of the substrate for the cultivation of the mushroom are given here as reference. (i) Sawdust 82%, wheat bran 16%, gypsum 1.4%, potassium phosphate, dibasic 0.2%, and lime 0.4%. (ii) Sawdust 54%, spent coffee grounds 30%, wheat bran 15%, and gypsum 1%. (iii) Sawdust 63%, corncob powder 20%, wheat bran 15%, calcium superphosphate 1%, and gypsum 1%. (iv) Sawdust 76%, wheat bran 18%, corn powder 2%, gypsum 2%, sugar 1.2%, calcium superphosphate 0.5%, and urea 0.3%. 2. Commercial-Scale Cultivation In general, the operation can use oak or other hardwood sawdust medium to grow the mushroom. The basic steps are (i) mix the sawdust, supplements, and water; (ii) bag the mixture; (iii) autoclave the bags to 121Ž C and cool the bags; (iv) inoculate and seal the bags; (v) incubate for 90 days to achieve full colonization of the sawdust mixture, in other words, to allow the mycelium to be established for ready fructification; (vi) fruit the colonized and established sawdust logs/bags/blocks 6 times using a 21-day cycle at 16–18Ž C; and (vii) harvest, clip steps, grade, box, and cold store for fresh market or harvest, dry, cut steps, grade, and dry again before boxing for dry market. The major equipment used in production consists of a mixer/conveyor, autoclave, gas boiler, cooling tunnel, laminar flow cabinet, bag sealer, air compressor for humidification, and shelves to incubate. Incubation can be done in two rooms and in two shipping containers. The two shipping containers can be installed near the fruiting rooms. The temperature during incubation is held between 18 and 25Ž C. Fruiting can be done in six rooms so that the blocks/logs can be moved as a unit. With compartmentalization, blocks in each room can be subjected to a cycle of humid cold, humid heat, and dry heat
MUSHROOM CULTIVATION 1 1.4.2.3 Cultivation of Pleurotus sajor-caju Pleurotus sajor-caju(grey er mushroo ngn-temperature sp od f This mushroom has a promising p ospect in tropical/subtropical areas.Its less complicated procedures (Chang and 1.Biological Nature s10-350 T The temperature for growth of myceli 24°.The the substrate used in making the mushroom bag/bed is 6.8-8.0.The C/N ratic in the substrate is in the range of 30:1-60: large circulation of air and reas ble ligh t are requre the development neat gra 57 or wheat bran 20%sugar 1%and gypsum 1%(vi Sawdust 58%spent coffee gent tea leaves 2 water hyacinthcereal straw 20.sugar 1.and Cultivation Substrate ()C tton seed h u95% gypsum 2 lime 1%. and lime 1.(Water hyacinth erea r gypsmnd lim 1%. For demonstration e.this mushroom can be nurtured to g ow into a tree like shape (Chang and Li,1982).The cultivation method,which has been tested t ssful,is as fol Cotton waste or nce straw mix ed with ater hyacint ter hvs Add 2% sm lin nd m with sufficient water to get moisture content of about 60-65%.Pile the materials ets,and leave to and overnight.Load the sub teurizat for 15 nd2w spawn thoroughly with the substrate and pack into columns of 60-cm-long tube which have hard plastic [polyvinyl chloride (PVC)]tubing of 100 cm(4 cm in dia upport and wi eets as ou mycelium of the mushroom has ramified the entire column of tre to four weeks.remove the plastic wrapping and switch on white light.Wateri d to eep the sur drying.In ound three to fou n period watering is very important if many flushes are required 1.4.2.4 Cultivation of Volvariella The edible straw mushroom volvariell volvacea is a fungus of the tropics and subtropics and has been traditionally culti vated in rice straw for many yeas in China and South East Asian countries.In 1971
MUSHROOM CULTIVATION 17 1.4.2.3 Cultivation of Pleurotus sajor-caju Pleurotus sajor-caju (grey oyster mushroom) is comparable to the high-temperature species in the group of Pleurotus (oyster) mushrooms, with high temperatures required for fructification. This mushroom has a promising prospect in tropical/subtropical areas. Its cultivation is easy with relatively less complicated procedures (Chang and Miles, 2004; Kaul and Dhar, 2007): 1. Biological Nature The temperature for growth of mycelium is 10–35Ž C. The optimum growing temperature of the mycelium is 23–28Ž C. The optimum developmental temperature of the fruiting body is 18–24Ž C. The optimum pH of the substrate used in making the mushroom bag/bed is 6.8–8.0. The C/N ratio in the substrate is in the range of 30 : 1–60 : 1. A large circulation of air and reasonable light are required for the development of the fruiting bodies. 2. Spawn Substrate (i) Wheat grain C 1.5% gypsum or lime. (ii) Cotton seed hull 88%, wheat bran 10%, sugar 1%, and gypsum 1%. (iii) Sawdust 78%, wheat bran 20%, sugar 1%, and gypsum 1%. (vi) Sawdust 58%, spent coffee grounds/spent tea leaves 20%, water hyacinth/cereal straw 20%, sugar 1%, and gypsum 1%. 3. Cultivation Substrate (i) Cotton seed hull 95%, gypsum 2%, lime 1%, and calcium superphosphate 2%. (ii) Rice straw 80%, cotton waste 18%, gypsum 1%, and lime 1%. (iii) Water hyacinth 80%, cereal straw 17%, gypsum 2%, and lime 1%. For demonstration purpose, this mushroom can be nurtured to grow into a treelike shape (Chang and Li, 1982). The cultivation method, which has been tested to be successful, is as follows: Cotton waste or rice straw mixed with water hyacinth is used as the substrate. Tear large pieces of cotton waste into small parts or cut the straw and water hyacinth into small segments. Add 2% (w/w) lime and mix with sufficient water to get moisture content of about 60–65%. Pile the materials up, cover with plastic sheets, and leave to stand overnight. Load the substrate into small baskets or on shelves for pasteurization or cook the substrate with boiled water for 15 minutes. After cooling to approximately 25Ž C, mix around 2% (w/w) spawn thoroughly with the substrate and pack into columns of 60-cm-long tubes which have hard plastic [polyvinyl chloride (PVC)] tubing of 100 cm (4 cm in diameter) as central support and with plastic sheets as outside wrapping. Incubate these columns at around 2428Ž C, preferably in the dark. When the mycelium of the mushroom has ramified the entire column of substrate after three to four weeks, remove the plastic wrapping and switch on white light. Watering occasionally is needed to keep the surface from drying. In around three to four days white primordia start to appear over the whole surface. After another two to three days, the Pleurotus mushrooms are ready for harvesting. During the cropping period watering is very important if many flushes are required. 1.4.2.4 Cultivation of Volvariella The edible straw mushroom Volvariella volvacea is a fungus of the tropics and subtropics and has been traditionally cultivated in rice straw for many yeas in China and South East Asian countries. In 1971
18 OVERVIEW OF MUSHROOM CULTIVATION AND UTILIZATION AS FUNCTIONAL FOODS cotton wastes were first introduced as heating material for growing the straw mush room(Yau and Chang.1972).and in 1973.cotton wastes had completely replaced the trad dy straw to grow the mushroom (Chang.1974) ind rial scale first in Hong Kong and then in Taiwan,Thailand,and elsewhere in Chin evera echnique s are adopte the 55 Detailed des ns of the various methods are iven by chang and Quimio (1982),Chang and Miles (2004).Kaul and Dhar (2007),and Quimio et al. (1990).Choice of te hnologies usually de son personal preference and the ty of sub or te the ind of the other technologies are low cost and app priate for rural area development. especially when production is established at the community level. 1.42.5 Cultivation of Agari us b ens In re nt years.A.brasili It ha and hishl nutitious mushroom but alsoan effective medicinal mushroom particularly for antitumour active polysaccharides. Agaricus asis was a wild mi the al, ere I d h ist Th cially was made in 1978.In 1992,this mushroom was introduced to China for commercial cultivation (Chang and Miles,2004). to middle-te mushrooms fom15t035 and the optimum growth temperature range is 23-27C.The temperature for and the e 2 in 60-75 70-85 for fruiting body formation and development.The optimum pH of the aking the mushroom bed is 6.5-6.8.The optimum pH of th A good circulation of requir evelopm of a his IInder natural conditions the mushr fo two crops each year.Each crop can harvest three flushes.According to the local climates,the far mer can decic de the spawning time in the year in order to have ms for harve h1n50 ays a kind Preparation om Bed (St Agaricus brasiliensis is strate rich in cellulose.The waste/b straw,wheat straw,bagasse (squeezed residue of sugar cane).cotton seed hull
18 OVERVIEW OF MUSHROOM CULTIVATION AND UTILIZATION AS FUNCTIONAL FOODS cotton wastes were first introduced as heating material for growing the straw mushroom (Yau and Chang, 1972), and in 1973, cotton wastes had completely replaced the traditional paddy straw to grow the mushroom (Chang, 1974). This was a turning point in the history of straw mushroom cultivation because the cotton waste compost through pasteurization brought the cultivation of the mushroom into an industrial scale first in Hong Kong and then in Taiwan, Thailand, and elsewhere in China. Several techniques are adopted for the cultivation of the mushroom, which thrives in the temperature range of 28–36Ž C and a relative humidity of 75–85%. Detailed descriptions of the various methods are given by Chang and Quimio (1982), Chang and Miles (2004), Kaul and Dhar (2007), and Quimio et al. (1990). Choice of technologies usually depends on personal preference and the availability of substrates and resources. While the more sophisticated indoor technology is recommended for the industrial-scale production of the mushroom, most of the other technologies are low cost and appropriate for rural area development, especially when production is established at the community level. 1.4.2.5 Cultivation of Agaricus brasiliensis In recent years, A. brasiliensis, formerly called Agaricus blazei Murill (Wasser et al., 2002), has rapidly become a popular mushroom. It has been proved to be not only a good-tasting and highly nutritious mushroom but also an effective medicinal mushroom, particularly for antitumour active polysaccharides. Agaricus brasiliensis was a wild mushroom in southeastern Brazil, where it was consumed by the people as a part of their diet. The culture of the mushroom was brought to Japan in 1965 and an attempt to cultivate this mushroom commercially was made in 1978. In 1992, this mushroom was introduced to China for commercial cultivation (Chang and Miles, 2004). 1. Biological Nature Agaricus brasiliensis belongs to middle-temperature mushrooms. The growth temperature for mycelium ranges from 15 to 35Ž C and the optimum growth temperature range is 23–27Ž C. The temperature for fruiting can be from 16 to 30Ž C and the optimum developmental temperature of fruiting bodies is 18–25Ž C. The ideal humidity for casing soil is 60–65%. The air humidity in a mushroom house is preferably 60–75% for mycelium growth and 70–85% for fruiting body formation and development. The optimum pH of the compost used in making the mushroom bed is 6.5–6.8. The optimum pH of the casing soil is 7.0. A good circulation of air is required for the development of the fruiting bodies. These conditions are similar to those needed for the cultivation of A. bisporus. Under natural conditions, the mushroom can be cultivated for two crops each year. Each crop can harvest three flushes. According to the local climates, the farmer can decide the spawning time in the year in order to have mushrooms for harvest within 50 days after spawning. 2. Preparation of Mushroom Bed (Stamets, 2000) Agaricus brasiliensis is a kind of mushroom belonging to straw-dung fungi and prefers to grow on substrate rich in cellulose. The waste/by-product agro-industrial materials [e.g., rice straw, wheat straw, bagasse (squeezed residue of sugar cane), cotton seed hull
MUSHROOM CULTIVATION 9 that ent e m poultry manure.and some chemical fertilizer.The following formulas for makin rence only.They should be modified according to the loca ns.(1)Rice straw /0%. cattle dun 0.5%,C0Cseed w1159 manure 15%calcium carbonate 1%,and ammonium sulfate or urea 0.5%.(i) Rice straw 90.%,rice bran 2.4%.fowl droppings 3.6%,slaked lime 19%,super and amm roppings 1.4.2.6 Cultivation of Ganoderma lucidum Although the medicinal value of G.lucidum has been treasured in China for more than 2000 years,the mushroom s lack c bility was largely respons the ng ancien ment and presented it to a high-ranking official was usually well rewarded (Chang and Miles. 2004) Artificia of th valuably mushroo was suc essfully oped rapidly Current the methods most wide production are the wood log.short wood segment.tree stump.sawdust bag.and bottle procedures Hsu,199 4:Mizuno et al..1996:Hung,1996:Mayzumi et al.. 1997;Chang and hich pawn directly under natural conditions.The third alternative technique involves the use of sterilized short logs about 12 cm in diameter and approximately 15 cm long which mycelial running.This meth 000-q0 nd duction procedure is more complex and the production costs much higher thar the natural log and tree stump meth ds.For this production procedure,the wood logs should be preparec 1D00 tre pre ng of t autumn and prior to the em ceof new le the following The onti mum moisture content of the log is about 45-55%.The flowchart for the short-log cultivation method is as follows:selection and felling of the tree,sawing/cutting th e log into short segmen g segme ins to pla ste development from the pinhead stage to maturity harvesting the fruiting bodie drying the fruiting bodies by electrical driers. ng.It should be noted that the prepared logs/segments are usually buried in soil inside a greenhouse or
MUSHROOM CULTIVATION 19 corn stalks, sorghum stover, and even wild grasses] can be used as the principal component of the compost for cultivation of the mushroom. It should be noted that these materials have to be air dried first and then mixed with cattle dung, poultry manure, and some chemical fertilizers. The following formulas for making compost are for reference only. They should be modified according to the local available materials and climatic conditions. (i) Rice straw 70%, air-dry cattle dung 15%, cottonseed hull 12.5%, gypsum 1%, calcium superphosphate 1%, and urea 0.5%. (ii) Corn stalks 36%, cottonseed hull 36%, wheat straw 11.5%, dry chicken manure 15%, calcium carbonate 1%, and ammonium sulfate or urea 0.5%. (iii) Rice straw 90.6%, rice bran 2.4%, fowl droppings 3.6%, slaked lime 1.9%, superphosphate 1.2%, and ammonium sulfate/urea 0.3%. (iv) Bagasse 75%, cottonseed hull 13%, fowl droppings 10%, superphosphate 0.5%, and slaked lime 1.5%. 1.4.2.6 Cultivation of Ganoderma lucidum Although the medicinal value of G. lucidum has been treasured in China for more than 2000 years, the mushroom was found infrequently in nature. This lack of availability was largely responsible for the mushroom being so highly cherished and expensive. During ancient times in China, any person who picked the mushroom from the natural environment and presented it to a high-ranking official was usually well rewarded (Chang and Miles, 2004). Artificial cultivation of this valuably mushroom was successfully achieved in the early 1970s and, since 1980 and particularly in China, production of G. lucidum has developed rapidly. Currently, the methods most widely adopted for commercial production are the wood log, short wood segment, tree stump, sawdust bag, and bottle procedures (Hsu, 1994; Mizuno et al., 1996; Hung, 1996; Mayzumi et al., 1997; Chang and Buswell, 1999; Stamets, 2000). Log cultivation methods include the use of natural logs and tree stumps which are inoculated with spawn directly under natural conditions. The third alternative technique involves the use of sterilized short logs about 12 cm in diameter and approximately 15 cm long which allow for good mycelial running. This method provides for a short growing cycle, higher biological efficiency, good-quality fruiting bodies, and, consequently, superior economical benefit. However, this production procedure is more complex and the production costs much higher than the natural log and tree stump methods. For this production procedure, the wood logs should be prepared from broad-leaf trees, preferably from oak. Felling of the trees is usually carried out during the dormant period, which is after defoliation in autumn and prior to the emergence of new leaves the following spring. The optimum moisture content of the log is about 45–55%. The flowchart for the short-log cultivation method is as follows: selection and felling of the tree, sawing/cutting the log into short segments, transfering segments to plastic bags, sterilization, inoculation, spawn running, burial of the log in soil, tending the fruiting bodies during development from the pinhead stage to maturity, harvesting the fruiting bodies, drying the fruiting bodies by electrical driers, and packaging. It should be noted that the prepared logs/segments are usually buried in soil inside a greenhouse or
20 OVERVIEW OF MUSHROOM CULTIVATION AND UTILIZATION AS FUNCTIONAL FOODS plastic shed.The soil should allow optimum conditions of drainage,air permeabil- ity,and water reter y should be avoide as containe note that the s us and can be modified according to the strains selected and the materials available in different localities):(i)sawdust 78%,wheat bran 20%,gypsum 1%,and soy gypsum I and cereal straw ash 1%(v)corn cob powder 78%.wheat/rice bran on beds nd f ang 1.4.3 Utilization of Mushroom Germplasm The item of mushroom germplasm is a selective subject only and is not an exhaus. and has to co success mush ooms and improv ng all desirable mushroom traits.It cannot be overempha sized that,to fully exploit the offered by mushrooms which have a co that could be quoted are the analytical chemists who analyze the many existing and rowth substrates used for mus 11n0 enzymes involved in the de adation of the individual components constitutin different substrates:fungal physiologists who focus their attention on the mecha ormation:gen eable to comprehend ms as strains with de who transter to the field scale the knowledge and technigues obtained in the laboratory. One of the ba breeding better quality mushroo y o arge re mushroom industry.Since all these phe pic differences are ultimately unde genetic control,mushr om strains with different traits actually possess distinctive an be ge ally by conv cro mushrooms them lves are the only source of this genetic material,the genes contained i existing mush ent the total g strain or uld me the tial loss of y thousands of unique g that could be used for breeding desirable new strains
20 OVERVIEW OF MUSHROOM CULTIVATION AND UTILIZATION AS FUNCTIONAL FOODS plastic shed. The soil should allow optimum conditions of drainage, air permeability, and water retention, but excessive humidity should be avoided. Examples of cultivation substrates using plastic bags or bottles as containers include the following (note that these examples are for reference purposes only and can be modified according to the strains selected and the materials available in different localities): (i) sawdust 78%, wheat bran 20%, gypsum 1%, and soybean powder 1%; (ii) bagasse 75%, wheat bran 22%, cane sugar 1%, gypsum 1%, and soybean powder 1%; (iii) cotton seed hull 88%, wheat bran 10%, cane sugar 1%, and gypsum 1%; (iv) sawdust 70%, corn cob powder 14%, wheat bran 14%, gypsum 1%, and cereal straw ash 1%; (v) corn cob powder 78%, wheat/rice bran 20%, gypsum 1%, and straw ash 1%. After sterilization, the plastic bags can be laid horizontally on beds or the ground for fruiting. 1.4.3 Utilization of Mushroom Germplasm The item of mushroom germplasm is a selective subject only and is not an exhaustive approach to mushroom utilization. It is concerned with the broadening of available mushroom resources in nature in order to conduct successful domesticating and breeding programs, with the aim at developing the cultivation of wild mushrooms and improving all desirable mushroom traits. It cannot be overemphasized that, to fully exploit the opportunities offered by mushrooms which have been properly collected and characterized, it is necessary to ensure a continuous exchange of information between scientists from different disciplines engaged in different areas of mushroom research. Included among the many possible examples that could be quoted are the analytical chemists who analyze the many existing and potentially new growth substrates used for mushroom cultivation in order to certify their suitability from an alimentary standpoint; biochemists who study the fungal enzymes involved in the degradation of the individual components constituting the different substrates; fungal physiologists who focus their attention on the mechanisms underlying carpophore formation; geneticists who are able to comprehend the life cycles of different mushrooms as well as to undertake breeding programs and select strains with desirable characteristics; and growers who transfer to the field scale the knowledge and techniques obtained in the laboratory. One of the basic requirements for breeding better quality mushrooms in higher yields is the wider availability of a large reserve of phenotypic variation (traits) which can be used for selection purposes by both researchers and the mushroom industry. Since all these phenotypic differences are ultimately under genetic control, mushroom strains with different traits actually possess distinctive gene combinations which can be generated artificially by conventional crossing methods, by protoplast fusion technology, and by transformation with genes cloned using recombinant deoxyribonucleic acid (rDNA) technology. Since the mushrooms themselves are the only source of this genetic material, the genes contained in existing mushroom strains and species represent the total genetic resource, that is, the entire pool of mushroom germplasm. Extinction of a single strain or species would mean the potential loss of many thousands of unique genes that could be used for breeding desirable new strains