1

UNI1LD NA1IONS - NA1IONS UNILS

LCONOMIC AND SOCIAL COMMISSION lOR ASIA AND 1lL PACIlIC


ASIAN AND PACIIIC CLN1RL IOR AGRICUL1URAL LNGINLLRING
AND MACHINLRY (APCALM)
A-7/I, CHINA IN1LRNA1IONAL SCILNCL AND 1LCHNOLOGY
CONVLN1ION CLN1RL
NO. J2, YUMIN ROAD, CHAOYANG DIS1RIC1, BLIJING J00029, P.R. CHINA








Training Manual on Mushroom Cultivation
Technology









1el: (86 J0) 822S 3S8J/3S80/3S78/3793 Iax: (86 J0) 822S 3S84 Lmail: infounapcaem.org
Website: www.unapcaem.org
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Acknowledgments

This report has been prepared by ProIessor Shu-Ting Chang, Emeritus ProIessor oI Biology oI
The Chinese University oI Hong Kong under contract with UNAPCAEM. Valuable comments
have been oIIered by a number oI persons Irom ProIessor G.Gantulga Irom Mongolia, Dr.
S.K.Adhikary Irom Nepal, Dr. Kwang-Jae CHOE Irom ROK, and ProIessor Ping Chang oI
UNAPCAEM. These comments have been incorporated where applicable.
Disclaimer

The designation used and the presentation oI the material in this publication do not
imply the express oI any opinion whatsoever on the part oI the Secretariat oI the United
Nations Economic and Social commission Ior Asian and the PaciIic (UNESCAP)
concerning the delimitation oI its Irontiers or boundaries.

The views expressed in this publication are those oI authors and do not necessarily
reIlect the view oI UNESCAP.

Mention oI Iirm names and commercial products does not imply the endorsement oI the
UNESCAP.
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Table of Content

ABSTRACT

1 INTRODUCTION

2. OVERVIEW OF THE MUSHROOM INDUSTRY (CONTENTS FOR A ONE-WEEK
MUSHROOM TRAINING WORKSHOP)

2.1 Mushrooms and Mushroom Biology
2.1.1 DeIinition oI a Mushroom
2.1.2 Mushroom Hunting
2.1.3 Ecological ClassiIication oI Mushrooms
2.1.4 Magnitude oI Mushroom Species
2.1.5 Concept oI Mushroom Biology

2.2 Applied Mushroom Biology
2.2.1 Mushroom Science---Food Supply through Mushroom Themselves
2.2.2 Mushroom Biotechnology ---Enhance Human Health through Mushroom Derivatives
2.2.3 Mushroom Bioremediation----BeneIit the Environment through Mushroom Mycelia

2.3 Mushroom Genetics and Breeding
2.3.1 Discovery oI Sexuality by Kniep and Bensaude
2.3.2 Sexuality in the Edible Mushrooms
2.3.3 Mating Systems (Patterns oI Sexuality) in Fungi
2.3.4 LiIe Cycle
2.3.5 Strain Improvement (Breeding Program)

2.4 Principle oI Mushroom Cultivation and Production
2.4.1 Mushroom Cultivation: Both a Science and an Art
2.4.2 World Mushroom Production
2.4.3 DiIIerences in Mushroom Production Patterns
2.4.4 World Mushroom Market
2.4.5 Development oI World Mushroom Movements (Organizations)

2.5 Mushroom Nutritional and Medicinal Properties
2.5.1 Nutritional Value oI Mushrooms
2.5.2 Medicinal Properties oI Mushrooms
2.5.3 Mushroom Nutriceuticals

3 MUSHROOM CULTIVATION TECHNOLOGY (CONTENTS FOR A MUSHROOM
TRAINING COURSE)

3.1 Major Phases oI Mushroom Cultivation
3.1.1 Selection oI An Acceptable Mushroom Species/Strains
3.1.2 Secreting a Good Quality oI Fruiting Culture
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3.1.3 Development oI Robust Spawn
3.1.4 Preparation oI Selective Substrate/Compost
3.1.5 Care oI Mycelial (Spawn) Running
3.1.6 Management oI Fruiting/Mushroom Development
3.1.7 Harvesting Mushrooms CareIully

3.2 Mushroom Diseases

3.3 Post-Harvest Handling

3.4 Cultivation oI several selected mushrooms
3.4.1 Agaricus bisporus (Champignon, Button mushroom)
3.4.2 Lentinula edodes (Xiang-gu, Shiitake, Oak mushroom)
3.4.3 Pleurotus safor-cafu (Grey oyster mushroom, Phoenix-tail mushroom, Indian oyster)
3.4.4 Jolvariella volvacea (Patty straw mushroom, Chinese mushroom)
3.4.5 Agaricus brasiliensis (Royal Sun Agaricus, Himematsutake)
3.4.6 Ganoderma lucidum (Ling Zhi, Reishi)

4 ENVIRONMENTAL IMPACT (SPECIAL TOPIC)

4.1 Reducing Environmental Pollution by Bioconversion oI Vast Quantities oI Organic Wastes
into Mushrooms

4.2 Recycling oI Vast Quantity oI Organic Wastes into Mushrooms, BioIertilizer and Biogas

4.3 Restoration oI Damaged Environment by Mushroom Mycelia

5 ECONOMIC AND SOCIAL IMPACTS (SPECIAL TOPIC)

6 CONCLUDING REMARKS

7 ACKNOWLEDGEMENTS

8 SUGGESTED FURTHER READINGS

9. ANNEX. RECONMENDATIONS FOR FOLLOW-UP ACTIVITIES

9.1 Problems Ior Cultivation oI Mushrooms in Some Countries in the Region

9.2 The Strategies Ior Developing Mushroom Industries in the Asian and PaciIic Region

9.3 Appropriate Training Ior Regional Mushroom Scientists and Mushroom Farmers

9.4 Consideration on the Formation oI a Regional Network on Mushroom Development


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ABSTRACT

1. Inadequate regional Iood supplies, diminishing quality oI health, and increasing
environmental deterioration are three key underlying problems aIIecting the Iuture well-being oI
humankind. The magnitude oI these problems is set to increase as the world`s population
continues to grow. The three Iacets oI Applied Mushroom Biology combined oIIer partial but
meaningIul solutions through (1) the generation oI relatively cheap source oI high quality Iood
protein (Mushroom Science), (2) the provision oI health-enhancing dietary supplements/
mushroom nutriceuticals (Mushroom Biotechnologv), and (3) the bioconversion/bioremediation
oI environmental adulterants and maintenance oI balanced ecosystems (Mushroom
Mvcorestoration).

2. Mushrooms are very nutritious products that can be generated Irom lignocellulosic waste
materials; and are in rich in crude Iibre and protein. In Iact, mushrooms also contain low Iat, low
calories and good vitamins. In addition, many mushrooms possess multi-Iunctional medicinal
properties.

3. Mushroom cultivation technology is Iriendly to the environment. The production oI edible and
medicinal mushrooms utilising, Ior example, paddy straw, cotton wastes, coIIee waste, water
hyacinth, tree saw dust, sugar cane bagasse, wild grasses and various categories oI reIuse and
lignocellulosic wastes, could readily be adopted in Asian and PaciIic communities in
sophisticated, but low technology approaches.

4. The spent substrate leIt aIter harvesting the mushrooms, which is entangled with innumerable
mushroom threads (collectively reIerred to as mycelia) will have been biochemically modiIied
by the mushroom enzymes into a simpler and more readily digestible Iorm, which is thus more
palatable to livestock, when used as a livestock Ieed supplement. Additionally, it will
signiIicantly have been enriched with protein, by virtue oI the remains oI the protein-rich
mycelia, leIt aIter harvesting the mushroom Iruiting bodies. The residue could also be utilised as
organic garden mulch, which is good Ior the soil.

5. Mushroom mycelia can produce a group oI complex extracellular enzymes which can degrade
and utilize the lignocellulosic wastes in order to reduce pollution. It has been revealed recently
that mushroom mycelia can play a signiIicant role in the restoration oI damaged environments.
Saprotrophic, endophytic, mycorrhizal, and even parasitic Iungi/mushrooms can be used in
mycorestoration, which can be perIormed in Iour diIIerent ways: mycoIiltration (using mycelia
to Iilter water), mycoIorestry (using mycelia to restore Iorests), mycoremediation (using mycelia
to eliminate toxic waste), and mycopesticides (using mycelia to control insect pests). These
methods represent the potential to create a clean ecosystem, where no damage will be leIt aIter
Iungal implementation.

6. The key objectives in each oI the participating countries will be, Iirstly, to develop Pleurotus
(oyster) mushrooms as additional, highly nutritious vegetable crops. We should start with these
because they are easiest to grow; yet they are also high in protein. Secondly, eIIorts will be
directed towards cultivating Lentinula mushrooms, which require less complicated and costly
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set-up and equipment (eg. compared with Agaricus mushrooms). Thirdly, eIIorts will be made to
produce selected medicinal mushrooms as dietary supplements, especially mushrooms which are
known to have a strong potency in invigorating the body`s immuno-response systems, such as
Ganoderma lucidum.

7. A comprehensive training programme Ior both researchers and mushroom growers will need
to be Iormulated. The identiIied team oI researchers will be brought together Ior an intensive
training course, which will equip them with more skills on how to train others, and also help
prepare them on how to succeed in this promising venture. Mushroom Iarming is both a science
and an art. The science, in its broad Iorm, will come through training workshops/courses. The art
will come through practical involvement and experience, and will have to be modiIied in
accordance with the prevailing conditions on site.

8. The potential oI mushroom Iarming in generating new employment opportunities is another
positive element emanating Irom mushroom Iarming ventures, which can be labour intensive.
Phase one oI this project will aim at providing initial skills Ior cottage level mushroom
production. Later, large scale development can be expected to lead to the establishment oI
commercial mushroom Iarms, and international marketing ventures. It is anticipated that
Governments oI the countries where Phase One oI Project implementation will be based, will
grant their political good-will, which will be a positive stimulus Ior private sector involvement.

9. Due to advances in both basic knowledge and practical technology relevant to mushroom
Iarming (mushroom themselves), mushroom products (mushroom derivatives) and mushroom
bioremediation (mushroom mycelia), these principles can be applied globally, but must be
implemented according to locally available substrates, labour and climatic conditions.


1. INTRODUCTION

Mushrooms are the Iruiting bodies oI macroIungi. They include both edible/medicinal and
poisonous species. However, originally, the word 'mushroom was used Ior the edible members
oI macroIungi and 'toadstools Ior poisonous ones oI the 'gill macroIungi. ScientiIically the
term 'toadstool has no meaning at all and it has been proposed that the term is dropped
altogether in order to avoid conIusion and the terms edible, medicinal and poisonous mushrooms
are used.

Edible mushrooms once called the 'Iood oI the gods and still treated as a garnish or delicacy
can be taken regularly as part oI the human diet or be treated as healthy Iood or as Iunctional
Iood. The extractable products Irom medicinal mushrooms, designed to supplement the human
diet not as regular Iood, but as the enhancement oI health and Iitness, can be classiIied into the
category oI dietary supplements/mushroom nutriceuticals (Chang and Buswell, 1996). Dietary
supplements are ingredients extracted Irom Ioods, herbs, mushrooms and other plants that are
taken without Iurther modiIication Ior their presumed health-enhancing beneIits.

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There is an old Chinese saying which states that ~MEDICINES AND FOODS HAVE A
COMMON ORIGIN. Mushrooms constitute a most rapidly growing new food category
which the current health-oriented public is increasingly enjoying.

Since mushrooms lack chlorophyll they can not, like green plants, get their energy Irom the sun
through photosynthesis. Instead, during their vegetative growth stage, mushroom mycelia secrete
enzymes that break down compounds such as cellulose and lignin present in the substrate. The
degraded compounds are then absorbed by the hyphae and the mycelium enlarges-usually
laterally, and in some cases growing several meters in diameter with the substrate.

Partially understood environmental Iactors (temperature and light are known to be critical)
stimulate the second or reproductive growth stage. Cells oI one mycelial strain Iuse with cells oI
the opposite type to Iorm a mycelium that contains both types oI nuclei. The new mycelium
continues to grow and eventually develops into a mature Iruiting body, the gills oI which are
lined with spore bearing cells called basidia. Various mechanisms trigger the dispersal oI spores,
which in turn lodge in a substrate, become hyphae and begin the cycle anew.

Mushroom cultivation has great scope in China, India and in some oI other developing countries
because oI the cheap and easily available raw materials needed Ior this activity, coupled with
Iaster means oI communication and marketing (as a Iresh commodity), and better purchasing
power oI the people. Using China as Ior example, in 1978, the production oI edible mushrooms
was only 60,000 tonnes. In 2006, China`s mushroom production was over 14 million tonnes.
Now there are more than 30 million people directly or indirectly engaged in mushroom
production and businesses, and now China has become a leading mushroom producer and
consumer in the world.

It is hoped that the avocation oI mushroom Iarming will become a very important cottage
industry activity in the integrated rural development programme, which will lead to the economic
betterment oI not only small Iarmers but also oI landless labourers and other weak sections oI
communities. The advantages oI mushroom cultivation can be summarized as:

1. Wastes such as cereal straws are largely burnt by the Iarmers, which causes air pollution.
However, these raw materials can actually be used Ior the cultivation oI mushrooms. This
kind oI bioconversion exercise can greatly reduce environmental pollution.
2. Mushroom cultivation can be a labour intensive activity. ThereIore, it will serve as
means oI generating employment, particularly Ior rural women and youths in order to
raise their social status. It will also provide additional work Ior the Iarmers during
winter months when the Iarming schedule is light.
3. It will provide the people with an additional vegetable oI high quality, and enrich the diet
with high quality proteins, minerals and vitamins which can be oI direct benefit to the
human health and fitness. The extractable bioactive compounds Irom medicinal
mushrooms would enhance human`s immune systems and improve their quality of
life.
4. Mushroom cultivation is a cash crop. The harvested Iruiting bodies can be sold in local
markets Ior additional Iamily income or exported Ior an important source oI Ioreign
exchange that will deIinitely improve the economic standards oI the people.
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5. Some warm mushrooms, e.g. Jolvariella volvacea (Straw mushrooms) and Pleurotus
safor-cafu (Oyster mushrooms) are relatively Iast growing organisms and can be
harvested in 3 to 4 weeks aIter spawning. It is a short return agricultural business and
can be of immediate benefit to the community.

Mushroom Iarming is both a science and an art. The science is developed through research the
art is perIected through curiosity and practical experience. However, mushroom Iarming is a
business which requires precision. Indeed, it is not as simple as what some people oIten loosely
stipulate. It calls Ior adherence to precise procedures. II you ignore one critical step, you are
inviting trouble, which could lead to a substantially reduced mushroom crop yield. For example,
iI you Iail to adjust the pH oI the substrate to a critical level required by the speciIic mushroom
species your are cultivating, or iI you do not properly pasteurize the substrate (to Iree the
mushroom spawn oI other moulds and various bacteria), your planted mushroom could be out-
competed by unwanted, intrusive micro-organisms.

The Iollowing ideas need to be emphasized as a conclusion oI this introduction. Although
scientiIic research and Iarming practice have led to the development oI some universal or general
concepts concerning mushroom cultivation, the diverse biological nature oI the process (in which
large numbers oI mushroom species and natural organic substrates are involved) also means that
a wide spectrum oI variations in Iarming methods must be employed. Thus the transIer oI
mushroom Iarming Irom one region or country to another cannot be treated in the same way as
the transIer oI non-biological industrial technology, such as that oI complete complex oI Iactory
equipment Ior textile or chemical Iertilizer industries. Since the cultivation oI mushrooms deals
with living organisms, one should consider, not only the unique attributes oI the mushroom itselI,
and oI the various micro-organisms growing with it (including both the harmIul and beneIicial
ones), but also the physical and biochemical natures oI the substrates. ThereIore, the cultivation
methods must be tailored in accordance with the prevailing unique natural resources, heritage,
local climate, and socio-economic conditions oI each Iarming community. All these
considerations call Ior a critical mass oI well trained mushroom scientists and mushroom
growers. Thus the training activities (workshops and courses) need to be supported by UN agents
as well as Governments oI the various countries concerned.


2. OVERVIEW OF THE MUSHROOM INDUSTRY (CONTENTS FOR A ONE-WEEK
MUSHROOM TRAINING WORKSHOP)


2.1 Mushrooms and Mushroom Biology

It has been well known that the 20th century has been an explosive time Ior the accumulation oI
knowledge. Modern technology Ior human civilisation is expanding every day. However, human
beings still Iace and will continue to Iace three basic problems: shortage oI Iood; pollution oI the
environment; and diminishing quality oI human health, due to the continued increase oI the
world population. The 20th century began with a world populated by 1.6 billion people and
ended with 6 billion inhabitants-- with most oI the growth occurring in the developing countries.
The growing world population is increasing by about 80 million people per year. At the present,
9
about 800 million people in the world are living in poverty. On the other hand, it has been
observed that over 70 oI agricultural and oI Iorest products has not been put to total
productivity, and have been wasted in processing. MacroIungi (mushrooms) not only can convert
these huge lignocellulosic biomass wastes into human Iood, but also can produce notable
immune enhanced products, which have many health beneIits. Another signiIicant aspect oI
mushroom cultivation is using the biota in creating a pollution-Iree environment.

2.1.1 Definition of a Mushroom

Mushrooms with other Iungi are something special in the living world, being neither plants nor
animals. They have been placed in a kingdom oI their own called the kingdom oI Myceteae. But
what are mushrooms? The word mushroom may mean diIIerent things to diIIerent people and
countries. It has emerged that specialised studies and the economic value oI mushrooms and their
products had reached a point where a clear deIinition oI the term 'mushroom was warranted. In
a broad sense 'Mushroom is a macroIungus with a distinctive Iruiting body, which can be either
epigeous or hypogeous and large enough to be seen with naked eye and to be picked by hand
(Chang and Miles, 1992). Thus, mushrooms need not be basidiomycetes, nor aerial, nor Ileshy,
nor edible. Mushrooms can be ascomycetes, grow underground, have a non-Ileshy texture and
need not be edible. This deIinition is not a perIect one but can be accepted as a workable term to
estimate the number oI mushrooms on the earth. The most common type oI mushrooms is
umbrella shaped with a pileus (cap) and a stipe (stem) i.e. Lentinula edodes. Other species
additionally have a volva (cup) i.e. Jolvariella volvacea or an annulus (ring) i.e. Agarius
campestris or with both oI them i.e. Amanita muscaria. Furthermore, some mushrooms are in the
Iorm oI pliable cups; others round like golI balls. Some are in the shape oI small clubs; some
resemble coral; others are yellow or orange jelly-like globs; and some even very much resembles
the human ear. In Iact, there is a countless variety oI Iorms.

The structure that we call a mushroom is in reality only the Iruiting body oI the Iungus. The
vegetative part oI the Iungus, called the mycelium, comprises a system oI branching threads and
cord-like strands that branch out through soil, compost, wood log or other lignocellulosic
material on which the Iungus may be growing. AIter a period oI growth and under Iavourable
conditions, the established (matured) mycelium could produce the Iruit structure which we call
the mushroom. Accordingly mushrooms can be grouped into Iour categories: (1) those which are
Ileshy and edible Iall into the edible mushroom category, e.g., Agaricus bisporus; (2)
mushrooms which are considered to have medicinal applications, are reIerred to as medicinal
mushrooms, e.g., Ganoderma lucidum; (3) those which are proven to be, or suspected oI being
poisonous are named as poisonous mushrooms, e.g., Amanita phalloides; and (4) a miscellaneous
category which includes a large number oI mushrooms whose properties remain less well
deIined, which may tentatively be grouped together as other mushrooms`. Certainly, this
approach oI classiIying oI mushrooms is not absolute and not mutually exclusive. Many kinds oI
mushrooms are not only edible, but also possess tonic and medicinal qualities.

Mushrooms are devoid oI leaves, and oI chlorophyll-containing tissues. This renders them
incapable oI photosynthetic Iood production. Yet, they grow, and they produce new biomass.
How? For their survival, Ior their growth, and Ior their metabolism, they rely on organic matter
synthesized by the green plants around us, including organic products contained in agricultural
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crop residues. The organic materials, on which mushrooms derive their nutrition, are reIerred to
as substrates. Mushrooms are a unique biota which assembles their Iood by secreting degrading
enzymes and decompose the complex Iood materials present in the biomass where they grow, to
generate simpler compounds, which they then absorb, and transIorm into their own peculiar
tissues. These substrate materials are usually by-products Irom industry, households and
agriculture and are usually considered as wastes. And these wastes, if carelessly disposed of in
the surrounding environment by dumping or burning, will lead to environmental pollution
and consequently cause health hazards. However, they are actually resources in the wrong
place at a particular time and mushroom cultivation can harness this waste/resource Ior its own
beneIicial advantage.

Mushrooms lack true roots. How then are they anchored into the substrates where we Iind them?
This is aIIected by their tightly interwoven thread-like hyphae, which also colonise the
substrates, degrade their biochemical components, and siphon away the hydrolysed organic
compounds Ior their own nutrition.

2.1.2 Mushroom Hunting

Fungi are Iound just about everywhere. Mushrooms, a special group oI macro-Iungi, are rather
more selective than other Iungi in that the size oI the Iruiting body requires the availability oI
more nutrients than are required Ior the production oI asexual spores by micro-Iungi. In damp
places, such as tree-Iern ecosystems and tropical rain Iorests, plentiIul moisture leads to
abundant mushroom Iormation. There, mushrooms can be collected at most times oI the year.
But in drier regions, they occur only aIter seasonal rains. In these ecosystems there may be a
particular Ilora oI mushroom species associated with the seasons oI autumn, summer and spring.
Relatively Iew mushrooms are produced during the cold winter months, although there are
perennial Iruiting bodies that persist during the winter. The Iormation oI mushroom Iruiting
bodies depends very much on the pattern oI rains and, in some years, there may be virtually total
lack oI mushroom Iruiting.

Mushroom hunters, in addition to carrying along with them the basic equipment and Iield guide
reIerences, which will vary depending on personal requirements and regional conditions, should
record such items as date, time, location, smell, substrate (host) colour, habitat and anything at
all unusual about the specimen. Some important characteristics Ior identiIication disappear
rapidly as the mushroom matures. These characteristics have to be recorded accurately at the
time oI collection.

2.1.3 Ecological Classification of Mushrooms

Ecologically, mushrooms can be classiIied into three groups: the saprophytes, the parasites and
the symbiotic (which include mycorrhizal) species (Fig. 1). There are only a Iew parasitic
mushrooms. Most oI the cultivated gourmet mushrooms are saprophytic Iungi. Some oI the
edible mushrooms are mycorrhizal species, e.g. Perigold black truIIle, Tuber melanosporum, and
matsutake mushroom, Tricholoma matsutake. It is diIIicult to bring these highly celebrated wild
gourmet species into cultivation because they are mycorrhizal. These species have a symbiotic
relationship with some vegetation, particularly trees, i.e. there is a relationship oI mutual need.
11

Saprophytes obtain nutrients Irom dead organic materials; parasites derive Iood substances Irom
living plants and animals and causing harm to the hosts; and mycorrhiza live in a close
physiological association with host plants and animals thereby Iorming a special partnership
where each partner enjoys some vital beneIits Irom the other.

However, some mushrooms do not Iall neatly within these man-made categories and can share
two oI these categories. For example, some Ganoderma spp. including G. lucidum are common
saprophytes, however they can be pathogenic too; also Tricholoma matsutake, while initially
appearing to be mycorrhizal on young roots, soon becomes pathogenic and Iinally exhibits some
saprobic ability.


Figure1. ModiIied triangular model Ior the ecological classiIication oI mushrooms.
Source: Hall et al., 2003.

2.1.4 Magnitude of Mushroom Species

In 1990, the number oI known species oI Iungi was about 69,000 (Hawksworth, 1991) while it
was conservatively estimated that 1.5 million species oI Iungi actually existed (Table 1). On
average 700 species were described as new to science each year Irom 1920 to 1950. However,
the annual total catalogued Iungi reached around 1,400 in 1961, 1,500 in 1968 and averaged
1,700 each year Ior 1986 to 1990.

Fungi are regarded as being the second largest group oI organisms in the biosphere aIter the
insects. Known Iungal species constitute only about 5 oI their species in the world. Thus, the
large majority oI Iungi are still unknown. Out oI about 70,000 described species oI Iungi, it has
been suggested that around 14,000-15,000 species produce Iruiting bodies oI suIIicient size and
Saprobe
Pathogen
MycorrhizaI
Cantharellus
cibarius
1richoloma
matsutake; 1ubur
melanosporum
Canoderma species
including C. lucidum
12
suitable structure to be considered as macroIungi (mushrooms). OI these, about 5,000 oI the
species are considered to possess varying degrees oI edibility, and more than 2,000 species Irom
31 genera are regarded as prime edible mushrooms. But only 100 oI them are experimentally
grown, 50 economically cultivated, around 30 commercially cultivated, and only about 6 to have
reached an industrial scale oI production in many countries. Furthermore, about 1,800 are
medicinal ones. The number oI poisonous mushrooms is relatively small (approximately 10),
oI these some 30 species are considered to be lethal (Miles and Chang, 1997).

Table 1. Comparison oI the numbers oI known and estimated total species in the world oI
selected groups oI organisms

Group Known species Total species known species

Vascular plants 220,000 270,000 81
Bryophytes 17,000 25,000 68
Algae 40,000 60,000 67
Fungi 69,000 1,500,000 5
Bacteria 3,000 30,000 10
Viruses 5,000 130,000 4

Source: Hawksworth (1991)

2.1.5 The Concept of Mushroom Biology

The biological science that is concerned with Iungi is called mycology. Mushroom biology is the
branch oI mycology that deals with mushrooms in many disciplines. When knowledge increases
and areas oI specialisation develop within the discipline, it is convenient to indicate that area oI
specialisation with a selI-explanatory name. In biology, there are such specialisations as
neurobiology, bacteriology, plant pathology, pomology, molecular biology, virology, Iungal
physiology, embryology, endocrinology, phycology, and entomology. These names indicate
either a group oI organisms (e.g., bacteria, algae, and insects) and /or an approach to the study
(e.g., disease, development and physiology).

Although several terms Ior this important branch oI mycology that deals with mushroom have
been used, and each oI these has its merit, when we get down to the matter oI deIinitions, it
seems that there is a place Ior a new term. The new term is mushroom biology. Mushroom
biology is a new discipline concerned with any aspect oI the scientiIic study oI mushrooms, such
as: taxonomy; physiology; genetics; etc.

2.2 Applied Mushroom Biology

Applied mushroom biology is concerned with all aspects oI the application oI mushroom
biology. It consists oI three main components: mushroom science; mushroom biotechnology; and
mushroom mycorestoration (Fig. 2). As previously outlined, mushroom biology is concerned
with any aspect oI the scientiIic study oI mushrooms, thereIore it will Ieature in each oI the three
components oI applied mushroom biology.
13


Figure2. The three components oI Applied Mushroom Biology: Mushroom Science; Mushroom
Biotechnology; and Mushroom Mycorestoration. Source: Chang and Buswell (2008).

Mushroom science deals with mushroom cultivation and production (mushrooms themselves)
and encompasses the principles oI mushroom biology/microbiology, bioconversion/composting
technology and environmental technology (Fig. 3).

Mushroom
Science
Mushroom
Mycorestoration
APPLIED
MUSHROOM
BIOLOGY
Mushroom
Biotechnology
14

Figure3. Mushroom science: mushroom cultivation and production. Source: Chang and
Buswell (2008).

Mushroom biotechnology is concerned with mushroom products (mushroom derivatives) and
encompasses the principles oI mushroom biology/microbiology, Iermentation technology and
bioprocess (Fig. 4). Mushroom biotechnology, both as a technology and as the basis Ior new
mushroom products, requires industrial development. It, like many bioscience industries,
operates at the cutting edge oI science and involves numerous regulatory issues.
Mushroom
Biology
MUSHROOM
SCIENCE
Compost
Technology
Environmental
Technology
Production of
mushrooms for
food
15


Figure 4.Mushroom biotechnology: mushroom products (mushroom nutriceuticals/dietary
supplements) Ior health enhancement. Source: Chang and Buswell (2008).

The third component oI applied mushroom biology has been developed in recent years. This is
mushroom bioremediation which is concerned with the beneIicial impacts oI mushrooms on the
environment (Irom mushroom mycelia) and encompasses principles oI mushroom
biology/microbiology, ecology and bioremediation technology (Fig. 5).


Mushroom
Biology

Bio-process
MUSHROOM
BIOTECHNOLOGY
Fermentation
Technology
Mushroom
derivatives for
health
16


Figure5. Mushroom mycorestoration: the beneIicial impacts oI mushrooms on the environment.
Source: Chang and Buswell (2008).

ThereIore, the aims oI the discipline oI applied mushroom biology are to tackle the three basic
problems: shortage oI Iood, diminishing quality oI human health and pollution oI the
environment, which human beings still Iace, and will continue to Iace, due to the continued
increase oI the world population. The 20
th
century began with a world population oI 1.6 billion
and ended with 6 billion inhabitants. The world`s population is likely to reach 9.2 billion in 2050
Irom the current 6.7 billion with most oI the growth occurring in developing countries. The
growing world population is increasing by about 80 million people per year. At present, about
800 million people in the world are living in poverty. On the other hand, it has been observed
that over 70 oI agricultural and oI Iorest products have not been put to total productivity, and
have been discarded as waste. Applied mushroom biology not only can convert these huge
lignocellulosic biomass wastes into human Iood, but also can produce notable nutriceutical
products, which have many health beneIits. Another signiIicant aspect oI applied mushroom
biology is using the biota in creating a pollution-Iree and beneIicial environment. The three
components oI applied mushroom biology are closely associated with three aspects oI wellbeing
- Iood, health and pollution.

2.2.1 Mushroom Science: Food Supplies through Mushroom Themselves

The discipline that is concerned with the principles and practice oI mushroom cultivation is
known as mushroom science. The establishment oI principles requires Iacts, which are arrived at
through systematic investigation. The systematic investigation must involve the practical aspects
oI mushroom cultivation as well as scientiIic studies. The consistent production oI successIul
mushroom crops necessitates both practical experience and scientiIic knowledge (see Section 3
on mushroom cultivation technology).
Mushroom
Biology

Bioremediation
Technology

MUSHROOM
MYCORESTORATION

Ecology
Beneficial
environmental
effects
17

2.2.2 Mushroom Biotechnology: Enhance Human Health through Mushroom Derivatives

It has been pointed out that mushroom biotechnology is concerned with mushroom products and
encompasses the principles oI Iermentation technology, mushroom biology/microbiology and
bioprocess. Mushroom products have a generalized or tonic eIIect, which in some cases may act
prophylactically by increasing resistance to disease in humans Irom the balancing oI nutrients in
the diet and the enhancing oI the immune systems.

(1) Nutritional value oI mushrooms

The greatest diIIiculty in Ieeding man is to supply a suIIicient quantity oI the body-building
material -- protein. The other three nutritional categories are: the source oI energy Iood
carbohydrates and Iats; accessory Iood Iactors-- vitamins; and inorganic compounds which are
indispensable to good health. OI course, water, too, is essential.

In terms oI the amount oI crude protein, mushrooms rank below animal meats, but well above
most other Ioods, including milk, which is an animal product (Chang and Miles 1989).
Furthermore, mushroom protein contains all the nine essential amino acids required by man.

The moisture content oI Iresh mushrooms varies within the range 70 - 95 depending upon the
harvest time and environmental conditions, whereas it is about 10 - 13 in dried mushrooms.

In addition to their good proteins, mushrooms are a relatively good source oI the Iollowing
individual nutrients: Iat, phosphorus, iron, and vitamins including thiamine, riboIlavin, ascorbic
acid, ergosterine and niacin. They are low in calories, carbohydrates and calcium. Mushrooms
also contain a high proportion oI unsaturated Iat.

In recent years, there has been a trend toward discovering ways oI treating mushrooms so as to
give them added value. For example, Wermer and Beelman (2002) have reported on growing
mushrooms enriched in selenium.

The desirability oI a Iood product does not necessarily bear any correlation to its nutritional
value. Instead, its appearance, taste, and aroma, sometimes can stimulate one`s appetite
(preIerence). In addition to nutritional value, mushrooms have some unique colour, taste, aroma,
and texture characteristics, which attract their consumption by humans.

(2) Medicinal value oI mushrooms

The second major attribute oI mushrooms, their medicinal properties, has also been drawn to our
attention Ior study, e.g., hypotensive and rental eIIects (Tam et. al., 1968; Yip et al., 1987),
immunomodulatory and antitumour activities oI polysaccharide-protein complex (PSPC) Irom
mycelial cultures (Liu et al., 1995, 1996; Wang et al., 1995, 1996, Ng et al., 1999),
immunomodulatory and antitumour activities oI lectins Irom edible mushrooms (Wang et al.,
1995, 1996, 1997), isolation and characterization oI a Type I Ribosome-Inactivation protein Irom
18
J. volvacea (Yao et al., 1998), and medicinal eIIects oI Ganoderma lucidum (Chang and Buswell
1999, Chang and Miles, 2004).

(3) Nutriceuticals and dietary supplements

The recent upsurge oI interest in traditional remedies Ior various physiological disorders and the
recognition oI numerous biological response modiIiers in mushrooms have led to the coining oI
the term 'mushroom nutriceuticals (Chang and Buswell, 1996). A mushroom nutriceutical is a
reIined/partially deIined mushroom extractive which is consumed in the Iorm oI capsules or
tablets as a dietary supplement (not a Iood) and which has potential therapeutic applications. A
regular intake may enhance the immune responses oI the human body, thereby increasing
resistance to disease and, in some cases, cause regression oI a disease state.

For more detailed coverage oI the nutritional and medicinal value and Ior comprehensive lists oI
mushrooms used in dietary supplements and in medicines, readers are reIerred to Section 2.5.

2.2.3 Mushroom Bioremediation: Benefit the Environment through Mushroom Mycelia

This component oI applied mushroom biology deals mainly with the aspects oI beneIits to the
Earth Irom the activities oI mushroom mycelium. Environmental contamination can be
ameliorated by the application oI mushroom mycelial technologies. For example, (1) the use oI
bioconversion processes to transIorm the polluting substances into valuable IoodstuIIs, e.g., the
proper treatment and reutilization oI spent substrates/composts in order to eliminate pollution
problems (Beyer, 2005, Noble, 2005). One oI the most intriguing opportunities oIIered by
mushroom mycelia in the area oI bioconversion is the exploitation oI their ability to degrade
pollutants, many oI which are highly carcinogenic, released into the environment as a
consequence oI human activity. And (2) the use oI Iungi/mushroom mycelia as tools Ior healing
soil, what Stamets (2005) called 'mycorestoration, which is the use oI Iungi/mushrooms to
repair or restore the weakened or damaged biosystems oI environment. The processes oI
mycorestoration include the selective use oI mushrooms Ior mycoIiltration, to Iilter water;
mycoIorestry, to enact ecoIorestry policy; mycoremediation, to denature toxic wastes; and
mycopesticides, to control insect pests. Mycoresoration recognizes the primary role
Iungi/mushrooms can play in determining the balance oI biological populations.

2.3 Mushroom Genetics and Breeding

Studies oI the genetics oI Iungi have played a leading role in the development oI the modern era
oI molecular genetics. The chemical studies oI Beadle and Tatum beginning in the 1940`s on the
biochemical genetics oI the ascomycete Neurospora brought Iorth techniques that were
subsequently employed with bacteria. Those studies then raised questions concerning the nature
oI the genetic material which culminated in the elucidation oI the structure oI DNA by Watson
and Crick in 1953. Since that time, Iundamental investigations on the molecular level have
progressed with ever-increasing rapidity.

The basidiomycetes were not in the Iore-Iront oI research during the early days when molecular
biologists preIerred the rapidity oI the cell cycle oI the unicellular bacterium Escherichia coli to
19
the slower growth oI the Iilamentous basidiomycetes with their complex incompatibility control
systems oI sexuality. In recent years, however, basidiomycetes have attracted the interest oI
investigators concerned with problems oI diIIerentiation and the control oI development because
they are eukaryotic organisms and do have some distinct but simple morphological stages which
are not present in single cell organisms. However, mushroom (macroIungus) genetics is a young
discipline in science as compared to biotechnology, which is correlated with the beginning oI
human civilisation. It was almost exclusively devoted to Iundamental research beIore attracting
serious attention during the early 1770s in relation to mushroom production.

The overall objective oI breeding is to improve the quality oI extant strains in the most eIIicient
way possible through the target selection, and controlled crossing, and progeny selection. The
desired result should be deIined with reasonable clarity in terms oI measurable traits desired,
such as yield, quality oI Ilavour, texture, appearance oI mushrooms, disease resistance, and
general vigour, all within the context oI a chosen set oI standard conditions which are employed
consistently throughout the breeding programme.

2.3.1 Discovery of Sexuality by Kniep and Bensaude

Although Blakeslee in 1904 had demonstrated sexuality in the Mucorales (class Zygomycetes)
through matings oI mycelia established Irom single spores, it was not until 1918-20 that the
mechanism oI sexuality in the basidiomycetes was discovered. It was demonstrated that the
mycelium arising Irom single spores was made up oI hyphae with simple septa, and that the
hyphae oI the Iruiting bodies and oI mycelium giving rise to Iruiting bodies bore clamp
connections. Furthermore, when mycelia which had originated Irom single spores were
conIronted with various combinations, only certain combinations gave rise to clamped
mycelium, while in other combinations the hyphae had simple septa.

2.3.2. Sexuality in the Edible Mushrooms

Although the process oI sexuality is complicated by nutritional and physiological conditions,
genetic constitution is the most critical Iactor determining both the occurrence and the
morphology oI the Iruiting bodies in the edible mushrooms.

Sexuality in Iungi consists oI three important stages. The Iirst essential stage is plasmogamy
which is the Iusion oI cytoplasm oI the two mating individuals. By plasmogamy the nuclei Irom
two strains are brought together in a common cytoplasm. The second essential stage in sexuality
is known as karyogamy or nuclear Iusion. The third essential stage is meiosis, the nuclear
division in which the chromosome number is reduced Irom the diploid to the haploid number.
The product oI meiosis is the Iormation oI a tetrad. Through the process oI sexuality, genetic
recombination and segregation subsequently occurs.

2.3.3 Mating Systems (Patterns of Sexuality) in Fungi

Edible mushrooms contain both selI-Iertile and selI-sterile species. SelI-Iertilisation
(homothallism or homomixis) is probably the commonest mode oI sexual reproduction in the
Iungi as a whole but in the higher Iungi (e.g., basidiomycetes) selI-Iertile species are in a distinct
20
minority oI only about 10 . SelI-sterilisation (heterothallism or dimixis) is about 90 , in
which 25 is bipolar and 65 is tetrapolar.

(1). Homothallism. Fruiting structure can be produced by a single, monosporous mycelium.
Potentially selI-Iertile Iungi are not always necessarily homozygous and a variety oI situation
and rather imprecise regulating system can result in heterozygosity. Two types oI homothallism
are Iound among selI-Iertile species: (a) primary homothallism, in which a homokaryotic
mycelium, established Irom a single meiotic nucleus, has the potentiality to progress through
heterokaryosis to the completion oI the sexual cycle; and (b) secondary homothallism, in which a
Iertile dikaryotic mycelium is established Irom a basidiospore carrying two meiotic nuclei oI
diIIerent mating types.

(2). Heterothallism. Cross mating between homokaryotic mycelia is required to complete the
sexual cycle. Two mating systems oI heterothallism are commonly Iound in edible mushrooms:
(a) Bipolar mating system, in which the mating competence is determined by incompatibility
Iactors oI a single series, the A Iactor. ThereIore, only two mating types are produced in equal
Irequency by a single Iruiting body. (b) Tetrapolar mating system, in which the mating
competence is determined by incompatibility Iactors oI two series, A and B, which assort and
segregate independently at meiosis. There are Iour rather than two mating types produced in
equal Irequency by a single Iruiting body.

2.3.4 Life Cycle

II a section oI the gills is cut and examined under the microscope, spores will be observed on
their surIace. The spores will start to Iall as the cap Iully expands, indicating maturity oI the
mushroom. The spores are so minute that they Iloat in the air and are carried by the wind.
Eventually, they Iall to the ground, usually with rain. II conditions are Iavourable (optimum
temperature and moisture), the spores will germinate to Iorm a mass oI mycelium. This is the
start oI the vegetative phase oI the mushroom. Given an unrestricted amount oI nutrients and
Iavourable growing conditions, it is capable oI unlimited growth. The mycelium developing Irom
the germinating spore is the so-called primary mycelium and is usually uninucleate and haploid.
This stage is short-lived because mycelia Irom diIIerent spores tend to ramiIy and Iuse to Iorm
the secondary mycelium with two compatible nuclei, which continues to grow vegetatively and
is able to Iorm Iruiting bodies.

2.3.5 Strain Improvement (Breeding Programme)

The reservoir oI edible mushrooms, like other micro-organisms oI use Ior industrial purposes, is
not unlimited. It is generally recognised that in order to maintain and breed high-yielding strains,
the techniques employed in mushroom breeding should now and then be modiIied and improved
in accordance with new Iindings and progress in the scientiIic world as a whole, and in
microbiology and genetics in particular.

(1).By selection. The common white mushroom, Agaricus bisporus, strains are usually based on
selection Irom multi-spore or single-spore cultures. Tissue culture oI selected sporophores has
also been used to Iix desired variants. In the short term, selection alone appears to have some
21
role to play in mushroom strain improvement, but genetic improvement by selection becomes
increasingly diIIicult. ThereIore, strain improvement through hybridisation has become a
recourse and also necessity.

(2). By hybridisation. In addition to the conventional method oI matings between two genetically
compatible strains through which dikaryon mycelia and Iruiting bodies are Iormed, steps towards
a broader spectrum oI hybridisation can be achieved in strain improvement oI edible mushrooms
by the Iollowing ways.

(A) Use oI auxotrophs. Auxotrophs can be obtained naturally or induced by mutagenesis.
The contrasting auxotrophs can be paired and the products can be screened Ior hybridity on
minimal medium. Certainly, the Ieasibility oI auxotrophs to be used as a tool Ior hybridisation
depends on how easily auxotrophs can be obtained in the strains oI the mushroom.

(B). Use oI resistance markers. Mutants resistant to antimetabolites have been suggested
recently as alternatives to auxotrophs Ior use in mushroom breeding programmes. The treated
spores or hyphal Iragments, which can grow on a medium containing an inhibitory concentration
oI the anti-metabolite, would be considered to possess the marker. Complementary resistant
strains would be grown together and, hybridity can be conIirmed by transIerring it to a medium
containing the two appropriate antimetabolites.

(C). Protoplast Iusion. One oI the most eIIective barriers to sexual reproduction is the
inability oI hyphae Irom two selected strains to Iuse. Several laboratories have reported that
protoplasts can be isolated Irom plant and microbial cells by enzymatic breaking oI the cell wall,
in the presence oI an osmotic stabiliser. Such protoplasts can be eIIectively induced to Iuse in the
presence oI polyethylene glycol (PEG). AIter a short period oI time protoplasts can regenerate
their cell walls and start to propagate as normal cells or hyphae. These cells are heterokaryotic iI
Iusion occurred between cells Irom genetically diIIerent strains. This can serve to increase the
Irequency oI intraspecies crosses in organisms in which natural matings rarely occur. The
technique has even a much wider application, and can be used Ior interspecies and intergeneric
crosses in some organisms, which normally cannot be crossed. Although such an approach has
been carried out in several laboratories, until now, no clear and economically applicable results
have been reported in edible mushrooms. I hope that this does not embrace the truth that
'beautiful hvpotheses are destroved bv uglv facts`. More detailed inIormation on mushroom
genetics and breeding can be Iound in 'Mushroom Biology--Concise basics and current
developments by Miles and Chang (1998) pp 65-85.

Due to the nature and the limited time oI the training course, the molecular biology oI
mushrooms was not touched upon in this course, although it has become an important sector oI
mushroom genetics and breeding. For Iurther reading, readers are reIerred to Chang et al (1993).

It should be emphasised that sexuality in any species has to be clearly understood in order to
conduct rational genetic studies as well as breeding programmes. Mushrooms representing three
types with respect to liIe cycle and genetic controls Ior mating have been considered. It is hoped
that the examples discussed in this course will provide some basic and useIul guidelines Ior
procedures as they might be applied generally to any breeding programme Ior each type oI
22
mushroom. Although it seems likely that selection Irom existing commercial strains oI edible
mushrooms may continue to enjoy some improvement, by analogy with other crop species,
hybridisation seems to oIIer the best prospects Ior real progress, especially with regard to
multiple gene transIers mediated by protoplast Iusion, during the next Iew decades. Finally, the
experience oI breeding in a variety oI organisms has demonstrated the wisdom oI preserving the
starting strains used in all breeding programmes Ior the purpose oI maintaining a pool oI
genetically diverse individuals. Such a practice would allow the possibility oI carrying out
additional breeding programmes to either improve or restore strains, which are in current use.

2.4 Principle of Mushroom Cultivation and Production

This section provides an overview oI mushroom cultivation and mushroom production. Detailed
inIormation on mushroom cultivation technology is in Section 3.

2.4.1 Mushroom Cultivation: Both a Science and an Art

The cultivation oI mushrooms can be both a relatively primitive Iarming activity, and a high
technology industry. In each case, however, continuous production oI successIul crops requires
both practical experience and scientiIic knowledge. Mushroom cultivation is both a science and
an art. The science is developed through research; the art is perIected through curiosity and
practical experience.

Mushroom growth dynamics involve some technological elements, which are in consonance with
those exhibited by our common agricultural crop plants. For example, there is a vegetative
growth phase, when the mycelia grow proIusely; and a reproductive (Iruiting) growth phase,
when the umbrella-like body that we call mushroom develops. In the agricultural plants, e.g.,
sunIlowers, when the plants switch Irom the vegetative growth to the reproductive growths,
retarded tips Ior Iurther growth (elongation) is an obvious phenomenon oI mature. It is the same
principle in mushroom production. AIter the vegetative (mycelial) phase has reached maturity,
what the mushroom Iarmer needs next is the induction oI Iruiting. This is the time the mycelial
growth tips should be retarded by regulating the environmental Iactors. These Iactors generally
called 'triggers or 'environmental shocks, such as, switching on the light, providing Iresh air,
and lowering temperatures, can trigger Iruiting (Fig. 6).













23




Triggers


Vegetative Phase Reproductive Phase


CO
2
O
2

Temp. Light













Figure 6.The two major phases oI mushroom growth and development - vegetative phase and
reproductive phase. The triggers Ior the transition Irom the vegetative phase to the reproductive
phase are usually regulated by environmental Iactors.

Although the principles oI cultivation are commonly similar Ior all mushrooms, the practical
technologies can be quite diIIerent Ior diIIerent species cultivated. The technologies have to be
modiIied and adjusted according to the local climatic conditions, materials available Ior
substrates and varieties oI the mushroom used.

2.4.2 World Mushroom Production

Generally, cultivated mushrooms should play a greater role in the endeavour to increase Iood
protein. This is especially true in developing countries, since growth substrates Ior mushrooms
are basically agricultural and industrial discards that are inedible Ior humans (Chang and Miles
1984). Biological (bioconversion) eIIiciency, i.e., the yield oI Iresh weight mushrooms in
proportion to the spawning compost in Agaricus or to the air-dried substrates in other non-
composting mushrooms, can reach 60-100 Ior Agaricus and 15-100 depending on the
cultivation conditions Ior other species.

24
The Iollowing statistics (Table 2) serve to illustrate dramatic increases in the production oI
Iarmed mushrooms during the period 1978 to 2006, with particular emphasis on China`s
contribution to total world production, given its current status as the leading mushroom producer
(Chang 1999, 2006b, Delcaire 1978; Sharma 1997):

Table 2.Total world mushroom production and China`s contribution since 1978
Year Total production
(x1,000 tonnes)
China`s production
(x1,000 tonnes)
China`s contribution
()
1978 1,060.0 60.0 5.7
1983 1,453.0 174.5 12.0
1990 3,763.0 1,083.0 28.8
1994 4,909.3 2,640.0 53.8
1997 6,158.4 3,918.0 63.6
2002 12,250.0 8,650.0 70.6
2006 n.a 14,000.0 n.a

Sources: Chang 1999, 2006b, Delcaire 1978; Sharma 1997 and courtesy communications with
the China Edible Fungi Association. n.a.: not available

Whereas in 1997, Asia contributed 74.4 oI the total world mushroom tonnage, Europe, 16.3
and North America, 7.0, both AIrica and Latin America`s shares were less than 1. This is
largely due to lack oI know-how, lack oI understanding that mushroom can play vital roles
towards enhancing human health when used as dietary Iood supplements, lack oI reliable sources
oI good quality mushroom spawn Ior supporting the eIIorts oI local mushroom growers, lack oI
venture capital to support mushroom Iarming entrepreneurs, and absence oI systematic
government support towards promoting mushroom Iarming as a valuable non-traditional new
Iood and cash crop (comparable to coIIee, tea, cotton, tobacco, etc.).

2.4.3 Differences in Mushroom Production Patterns

The mushroom industry in UK and in some other Western countries is oIten overwhelmingly
Iocused on one mushroom species, Agaricus bisporus. These industries are nearly 100
dominated by Agaricus bisporus (Gaze, 2005). In the US, it accounts Ior about 98 oI its
mushroom industry, Lentinula edodes Ior 1 and Pleurotus spp Ior only about 0.5 (Table 3).
However, it should be noted that Agaricus bisporus is only but one oI many edible Iungi
cultivated globally.
25

This production pattern is slightly less skewed towards Agaricus bisporus in Spain, the third
largest mushroom producer in the EU. In 2004, mushroom production in Spain was 110,000
tonnes compared with 26,512 tonnes in 1992, increasing 315. Production consists oI 80 oI
Agaricus mushrooms, 15 oI Pleurotus mushrooms and 5 oI Lentinula mushrooms.

Table 3. The US mushroom industry is dominated by Agaricus Bisporus

Mushroom Production (t) 02-03 03-04 04-05
Agaricus 379318.8 381479.4 380083.0
(98.6%) (98.4%) (98.2%)
Lentinula 3390.5 3506.1 4118.4
(0.9%) (0.9%) (1.1%)
Pleurotus 1812.6 2008.2 2453.0
(0.5%) (0.5%) (0.6%)
Sub-total 384521.9 386993.7 386654.4
(99.9%) (99.9%) (99.9%)
Others 197.3 541.0 253.5
(0.01%) (0.01%) (0.01%)
Grand total 384719.2 387534.7 386907.9
(100%) (100%) (100%)
Source: National Agricultural Statistics Service (NASS) 2005, USA.
Note: share oI total US mushroom production is in brackets.

On the other hand, specialty mushrooms in East Asian countries are Iar more popular than A.
bisporus as shown in Table 4. Agaricus accounted Ior 12.8 oI total mushroom production in
China in 2003, 11.6 in S. Korea and 0 in Japan. Furthermore, while the production oI the
three important mushrooms, Agaricus, Lentinula and Pleurotus mushrooms together make up
nearly 100 oI the mushroom industry in U.S and Spain, the production oI these three
mushrooms account Ior 72.7 oI total mushroom production in S. Korea, 58.2 Ior China and
only 12.3 in Japan (Table 4). This means there are more other culinary-medicinal mushrooms
being cultivated and marketed in those three Asian countries particularly in Japan.

Table 4. Asia`s mushroom industry is more diverse (2003 data)

Production China (x1000t) Japan (t) S. Korea (t) Taiwan (t)
Agaricus 1330.4 - 19790 4276
(12.8%) (11.6%) (4.0%)
Lentinula 2228 35294 41876 36000
(21.5%) (10.7%) (24.6%) (33.4%)
Pleurotus 2488 5219 61965 4540
(24.0%) (1.6%) (36.5%) (4.2%)
Sub-total 6046.4 40513 123631 44816
(58.2%) (12.3%) (72.7%) (41.6%)
Others 4340.5 290333 46369 62984
(41.8%) (87.8%) (27.3%) (58.4%)
Grand total 10386.9 330846 170000 107800
26
(100%) (100%) (100%) (100%)
Source: Chang, 2006; Cui, 2004; Ho and Peng, 2006
Note: shares oI total mushroom production are in brackets.

2.4.4 World Mushroom Market

The world market Ior the mushroom industry in 2001 was valued at over US$40 billion. The
mushroom industry can be divided into three main categories: edible mushrooms valued about
US$30 billion; medicinal mushroom products were worth about US$9 - 10 billion; and wild
mushrooms, US$4-5 billion. International bodies/Iorums have developed Ior each oI these
segments oI the mushroom industry that has helped to bring them to the IoreIront oI international
attention: 1) International Society oI Mushroom Science (ISMS), Ior edible mushrooms
established in 1950 in England; 2) World Society Ior Mushroom Biology and Mushroom
Products (WSMBMP), Ior mushroom biology and medicinal mushroom products Iormed in 1993
in Hong Kong; and 3) International Workshops on Edible Mycorrhizal Mushrooms, Ior some
wild mushrooms initiated in 1999 in Sweden. The three international bodies/Iorums (see next
section Ior Iurther details) have done much to promote each oI their respective Iields, not least oI
which is bringing scientists together Ior useIul discussions, encouraging research and the
dissemination oI valuable research and commercial inIormation.

The outlook Ior many oI the known mushroom species is bright. Production oI mushrooms
worldwide has been steadily increasing, mainly due to contributions Irom developing countries,
such as China, India, Poland, Hungary and Vietnam. There are also increasing experimentally
based evidence to support centuries oI observations regarding the nutritional and medicinal
beneIits oI mushrooms. The value oI mushrooms has recently been promoted to tremendous
levels with medicinal mushrooms trials conducted Ior HIV/AIDS patients in AIrica, generating
encouraging results. However, harvests oI highly prized edible mycorrhizal mushrooms are
continuously decreasing. This has triggered research into devising methods Ior improved
cultivation oI wild mushroom. It is hoped that there will be even more research into this area, so
that larger quantities oI wild mushrooms can be massively harvested through artiIicial cultivation
or semi-cultivation methods. Technological developments in the mushroom industry in general
has seen increasing production capacities, innovations in cultivation technologies, improvements
to Iinal mushroom goods, capitalising on mushrooms` nutritional and medicinal properties, and
utilising mushrooms` natural qualities Ior environmental beneIits.

2.4.5 Development of World Mushroom Industry Movements (Organizations)

Although mushrooms have been collected Irom the wild and cultivated artiIicially Ior human
Iood and Ior medicine uses Ior hundreds and thousands oI years, it is only recently that the three
main segments oI the mushroom industry could be identiIied. These three segments have
received international recognition as important inter-related components (Fig. 7), with each
deserving its own special patronage and paths oI development: (a) cultivated edible mushrooms
(mushroom themselves-used directly or indirectly as Iood); (b) medicinal mushrooms
(mushroom derivatives-used as nutriceutical therapy/dietary supplements); and (c) wild
mushrooms including edible mycorrhizal, symbiotic and poisonous mushrooms (collected, up to
now, only Irom the wild). The development oI three important international bodies/Iorums has
27
helped to bring each oI these three components oI the mushroom industry to the IoreIront oI
international attention, showcasing their positive contributions to human welIares (Chang,
2006b).

(1). The international movement Ior edible mushrooms, mainly concerned with mushroom
production (mushroom themselves), was initiated during the Iirst International ConIerence on
Mushroom Science held in Peterborough, UK, 3
rd
to 11
th
May 1950. Chairman F. C. Atkins with
P. J. Bels, E. B. Lambert and R. L. Edwards were on the organising committee. The committee
members later Iormed the International Commission on Mushroom Science which eventually
developed into the International Society Ior Mushroom Science (ISMS) (Personal
communication with Peter Flegg 2005).

The 17
th
International Congress oI ISMS will be held in May 2008 in Cape Town, South AIrica.
Traditionally, the Iocus oI the ISMS has been on the Agaricus bisporus mushroom industry. In
recent years, the interests oI the ISMS have become more diversiIied but A. bisporus is still its
main concern. The ISMSC-18 will be held in Beijing, China, in 2012.

(2). The international movement Ior medicinal mushrooms, mainly concerned with mushroom
products (mushroom derivatives), was instituted during the Iirst International ConIerence on
Mushroom Biology and Mushroom Products held in Hong Kong, 23-26 August, 1993. Chairman
S. T. Chang with J. A. Buswell, V.E.C. Ooi, K.W.K. Liu and S.W. Chiu were on the organising
committee.

The World Society Ior Mushroom Biology and Mushroom Products (WSMBMP) was launched
in January 1994 in response to strong interest expressed at the conIerence in Hong Kong in the
previous year. The object oI the WSMBMP is to promote the enhancement and application oI
knowledge related to the basic and applied aspects oI mushroom biology and mushroom
products (mushroom derivatives possessing medicinal properties Irom edible, medicinal and wild
mushrooms) through publications, meetings and other means deemed appropriate. The
WSMBMP holds a mushroom meeting, International ConIerence Ior Mushroom Biology and
Mushroom Products (the ICMBMP) every three years. The sixth one is to be held in September
2008 in Bonn, Germany.

The international movement Ior the medicinal segment oI the mushroom industry has been given
a Iurther boost with the launch oI the International Journal oI Medicinal Mushrooms (IJMM) in
1999 by Solomon P. Wasser as Editor-in-ChieI with Takashi Mizuno, Shu Ting Chang, and
Alexander L. Weis as editors. This then led to the inaugural International Medicinal Mushroom
ConIerence (IMMC) held in Kiev, Ukraine, 12-14 September, 2001. It has been agreed that there
is an IMMC aIter an interval oI two years. The 2
nd
IMMC was held in Pattaya, Thailand, 17-19
July 2003 and the 3
rd
was held in Port Townsend, Washington, USA, Irom 12 to 17 October
2005. IMMC 4 will be in Slovenia, 2007 and the IMMC 5 will be in Nantong, China, 2009.

(3). The international movement Ior wild mushrooms, mainly concerned with edible mycorrhizal
mushrooms, was born as a Pre-Congress activity during the 2
nd
International ConIerence on
Mycorrhizas in Uppsala, Sweden, in 1999. Two years later, the 2
nd
International Workshop on
Edible Mycorrhizal Mushrooms (IW-EMM) was held in Christchurch, New Zealand, 3-6 July
28
2001. The 3
rd
IW-EMM was hosted by the University oI Victoria, Canada, 16-22, August 2003
and the 4
th
was held in Murcia, Spain, 29 November to 2 December, 2005. The 5th IW-EMM
was held in Yunnan, China, 2007. It should be noted that edible mycorrhizal mushrooms belong
to a special group oI wild mushrooms which include other symbiotic mushrooms, e.g., termite
mushrooms, hallucinogenic, and poisonous mushrooms.

These three international bodies/Iorums have done much to promote each oI their respective
Iields, not least oI which is bringing together scientists in international Iorums Ior useIul
discussions, encouraging research and the dissemination oI valuable inIormation. These three
segments oI the mushroom-based industry are not Ior competition but Ior complementation.

2.5 Mushroom Nutritional and Medicinal Properties

Edible mushrooms provide high quality oI protein that can be produced with greater biological
eIIiciency than animal protein. They are rich in Iibre, minerals and vitamins, and have low crude
Iat content, with a high proportion oI polyunsaturated Iatty acids (72 to 85 ) relative to total
content oI Iatty acids. These properties are major contributing Iactors to the traditional
recognition oI mushrooms as 'healthy Ioods.

A large number oI mushroom species are not only edible and nutritious but also possess tonic
and medicinal qualities. However, some mushrooms are lethally poisonous, and one should eat
mushrooms only iI one knows their names and their properties with considerable precision.

In the past, the mushroom industry concentrated mainly on the production oI Iresh, canned and
dried mushrooms Ior Iood. Thus, the industry had only one leg. In the present era, high-pressure
work demands are causing greater stress to the human body, and resulting in the weakening oI
the human immune system. A variety oI proprietary products based on mushroom nutriceuticals
and mushroom pharmaceuticals have already been produced and marketed. This trend is
expected to increase with wider consumer satisIaction and acceptability. This is the second leg oI
the industry. These two legs/segments oI the mushroom-based industry will not compete but will
complement each other.

2.5.1 Nutritional Value of Mushrooms

The greatest diIIiculty in Ieeding man is to supply a suIIicient quantity oI the body-building
material -- protein. The other three nutritional categories are: the source oI energy Iood
carbohydrates and Iats; accessory Iood Iactors-- vitamins; and inorganic compounds which are
indispensable to good health. OI course, water, too, is essential.

The moisture content oI Iresh mushrooms varies within the range oI 70 - 95 depending upon
the harvest time and environmental conditions, whereas it is about 10 - 13 in dried mushrooms.
The protein content oI the cultivated species ranges Irom 1.75 to 5.9 oI their Iresh weight. It
has been estimated that an average value oI 3.5 to 4.0 would be more representative. This
means that the protein content oI edible mushrooms in general, is about twice that oI onion (1.4
) and cabbage (1.4), and Iour times and 12 times those oI oranges (1.0 ) and apples (0.3
), respectively. In comparison, the protein content oI common meats is as Iollows: pork, 9-16
29
; beeI, 12-20 ; chicken, 18-20 ; Iish, 18 -20 ; and milk, 2.9- 3.3 . On a dry weight basis,
mushrooms normally contain 19 -35 protein, as compared to 7.3 in rice, 12.7 in wheat,
38.1 in soybean, and 9.4 in corn. ThereIore, in terms oI the amount oI crude protein,
mushrooms rank below animal meats, but well above most other Ioods, including milk, which is
an animal product (Chang and Miles 1989). Furthermore, mushroom protein contains all the
nine essential amino acids required by man.

Quantitative data relating to the nutritive value oI mushrooms is sparse. In the absence oI Ieeding
trials, alternative methods have been used to determine or predict the nutritional value oI Ioods
based on their content oI essential amino acids (Crimson & Sands 1978).The Essential Amino
Acid Index (EA. Index) rates dietary protein in terms oI an essential amino acid pattern based on
known adult human dietary requirements. The Amino Acid Score (Chemical Score) is the
amount oI the most limiting amino acid in the Iood protein expressed as a percentage oI the same
amino acid present in the reIerence protein. In an attempt to resolve the diIIiculties inherent in
comparisons between those mushrooms containing small amounts oI high quality protein with
those containing larger amounts oI a protein oI lesser nutritional quality, Crisan & Sands (1978)
proposed the use oI a Nutritional Index calculated as:

(EAA index x percentage protein)
Nutritional Index ----------------------------------------------
100

The EAA Indices, the Amino Acid Scores, and Nutritional Indices Ior various mushrooms and
other Ioods, are reported by Crisan & Sands (1978). EAA Indices and Amino Acid Scores oI the
most nutritive mushrooms (highest values) rank in potential nutritive value with those oI meat
and milk, and are signiIicantly higher than those Ior most legumes and vegetables. The least
nutritive mushrooms rank appreciably lower but are still comparable to some oI our common
vegetables.

In addition to their good proteins, mushrooms are a relatively good source oI the Iollowing
individual nutrients: Iat, phosphorus, iron, and vitamins including thiamine, riboIlavin, ascorbic
acid, ergosterine and niacin. They are low in calories, carbohydrates and calcium. It has also
been reported that a total lipid content varying between 0.6 and 3.1 oI the dry weight, is Iound
in the commonly cultivated mushrooms. At least 72 oI the total Iatty acids are Iound to be
unsaturated in all the Iour tested mushrooms (Huang, et al., 1985). It should be noted that
unsaturated Iatty acids are essential and signiIicant in our diet and to our health.

In recent years, there has been a trend toward discovering ways oI treating mushrooms so as to
give them added value. For example, Wermer and Beelman (2002) have reported on growing
mushrooms enriched in selenium. By adding sodium selenite to compost over a range oI 30-300
parts per million, they Iound that the mushrooms increasingly absorbed selenium according to
the amount in the compost, so that it is possible to grow mushrooms containing a desired
concentration. Selenium is an essential micronutrient that has generated much recent interest in
nutritional and medical researchand, more recently, within the Iood industry (Beelman and
Royse, 2006). Selenium has numerous physiological Iunctions, but is best known as necessary
30
coIactor Ior the glutathione peroxidase enzyme system. This system is responsible Ior removing
Iree radicals Irom the body, thus reducing oxidative damage.

The desirability oI a Iood product does not necessarily bear any correlation to its nutritional
value. However, its appearance, taste, and aroma, sometimes can stimulate one`s appetite
(preIerence). In addition to nutritional value, mushrooms have some unique colour, taste, aroma,
and texture characteristics, which attract their consumption by humans.

2.5.2 Medicinal Properties of Mushrooms

The second major attribute oI mushrooms, their medicinal properties, has long been recognised
in China, Korea, and Japan. There has been a great upsurge in activities related to the use oI
mushroom products Ior medicinal purposes in recent years. In 2001, the Iigure oI US$9-10
billion was cited as representing the value oI medicinal mushroom products, including tonics and
medicines. The application oI modern analytical techniques can be used to establish a scientiIic
basis Ior the empirical observations that have been made centuries beIore. According to Chang
and Buswell (1996), the term 'mushroom nutriceutical is used Ior a new class oI new
compounds extractable Irom either the mycelium, or the Iruiting body oI the mushroom.
Mushroom nutriceuticals may possess both nutritional and medicinal properties.

Due to present day high pressured work demands resulting in great stress to the human body and
causing a weakening oI the human immune system, there are now many new diseases. These
have developed as a consequence oI lower natural body resistance. There is some evidence that
the beneIicial treatment oI these diseases can be obtained by consumption oI mushrooms as a
Iunctional Iood, or through the use oI extracted biologically active compounds as a dietary
supplement, in order to enhance immune response oI the human body, thereby increasing
resistance to disease and, in some cases, causing regression oI a diseased state. DiIIering Irom
most pharmaceuticals, these biologically active compounds extracted Irom medicinal mushrooms
have extraordinarily low toxicity, even at high doses. Long viewed as tonics, now it has been
known that they can proIoundly improve the quality oI human health.

Mushrooms produce several biologically active compounds that are usually associated with the
cell wall. Most notably, a group oI polysaccharides comprising high molecular weight sugar
polymers has been reported to contribute to their immune enhancing and tumour retarding
eIIects. It has been reported that the anti-tumour and anti-cancer eIIects oI the polysaccharides
are based on the enhancement oI the body`s immune systems, including activated macrophages,
natural killer cells, cytotoxic T cells, and their secretory products, such as the tumour necrosis
Iactor, reactive nitrogen and oxygen intermediates, and interleukins, rather than direct cytocidal
eIIects (Mizuno et al. 1995; Liu et al. 1996). It should be noted that immune responses are
complex reactions involving several types oI cells, such as macrophages and lymphocytes. The
killing mediated by cytotoxic T lymphocytes and natural killer cells, represents an important
mechanism in immune deIence against tumours, virus-inIected cells, parasites and other Ioreign
invaders. Another group oI medicinal compounds Iound in Ganoderma spp. are triterpenoids,
steroid-like compounds, which contribute cytotoxic, hepatoprotective, and hypolipidmic
inIluence on platelet aggregation, inhibition oI angiotensine-converting enzyme, and inhibition
oI histamine release (Lindequist 1995). Lectins, another group oI mushroom bioactive
31
compounds, are proteins or glycoproteins with speciIic binding sites Ior sugars, which are not
antibodies or enzymes. These have become useIul tools in structural studies oI the cell surIace,
oligosaccharides and/or carbohydrate moieties oI glycoproteins. Some lectins have been shown
to have anti-tumour and immunomodulatory activities (Wang et al. 1996). Other lectins
preIerentially agglutinate mammalian cells that have been transIormed by oncogenic viruses or
by chemical carcinogens, as well as spontaneously transIormed cells. These and related Iindings
indicate that studies with lectins may lead to a better understanding oI cancer. Moreover, some
lectins may be used to inhibit the growth oI malignant cells. A novel single-chained ribosome-
inactivating protein (RIP) was recently isolated Irom Iruit bodies oI the edible mushroom, J.
volvacea (Yao et al. 1998). The mushroom RIP, designated volvarin, exhibited a potent
inhibitory action on protein synthesis in the rabbit reticulocyte lysate system. Like most plant
RIPs, volvarin acted as an N-glycosidase that depurinated rRNA Irom rabbit reticulocyte lysate,
releasing a characteristic RNA Iragment aIter treatment with aniline. It also exerted a strong
abortiIacient eIIect in mice.

Furthermore, the aqueous extracts oI Pleurotus safor-cafu (Tam et al. 1986) and Jolvariella
volvacea (Chiu et al. 1996) have been reported to produce a hypotensive eIIects in normotensive
rats. Feeding powdered maitake (Grifola frondosa) mushrooms to spontaneous hypertensive rats
resulted in a lowering oI the blood pressure (Kyoko et al. 1988). It has also been reported that
dried powder oI another two edible mushrooms, Auricularia auricula and Tremella fuciformis,
aIter being Ied to the rats, has demonstrated to be eIIective in lowering both the serum total
cholesterol and the low density lipoprotein (LDL) cholesterol levels (Cheung 1996). Since the
mushrooms did not aIIect the concentration oI serum high density lipoprotein (HDL) cholesterol,
the reduction oI serum total cholesterol by the mushroom diets is believed to be attributable to
the Iall in LDL cholesterol. It should be noted that LDL is'bad cholesterol and HDL is 'good
cholesterol.

2.5.3 Mushroom Nutriceuticals

There has been a recent upsurge oI interest in mushrooms not only as health vegetables (Iood)
but also as a source oI biological active compounds oI medicinal value, including use as
complementary medicine/dietary supplements Ior anticancer, antiviral, immunopotentiating,
hypocholesterolaemic and hepatoprotective agents. This new class oI compounds termed
mushroom nutriceuticals` (Chang and Buswell 1996), are extractable Irom either the Iungal
mycelium or Iruiting body and represent an important component oI the expanding mushroom
biotechnology industry.

OI the 14,000-15,000 species oI so-called mushrooms in the world, around 400 have known
medicinal properties. However, it has been estimated that there are about 1,800 species oI
mushrooms with the potential oI medicinal properties. Both these mushrooms and their root-like
structure (called mycelium) produce several medicinal or nutriceutical (general immune
enhancing) compounds, central oI which are the polysaccharides (high molecular weight strings
oI sugars), triterpenes, and immunomodulatory proteins. Although virtually all mushrooms and
many Ioods have polysaccharides in their cell walls, certain mushroom species have been Iound
to contain polysaccharides which are particularly eIIective in retarding the progress oI various
cancers and other diseases, and in alleviating the side eIIects oI chemotherapy and radiation
32
treatment (through cell-level regenerative eIIects). There are now many studies in Asia,
particularly in China and Japan, documenting liIe span increases oI cancer patients undergoing
conventional cancer treatment plus mushroom extract consumption or injection (Mizuno et al.,
1995; Liu 1999). At the same time, due to the enhancement oI the immune systems, it can help
people reduce the possibility oI being inIected by other diseases.

Between 80 - 85 oI all medicinal mushroom products are derived Irom the Iruiting bodies,
which have been either commercially Iarmed or collected Irom the wild, e.g., Lentinan, a high-
molecular weight (1-3)--D-glucan, Irom Lentinula edodes and various products Irom
Ganoderma lucidum. Only about 15 oI all products are based on extracts Irom mycelia. The
notable examples include PSK-trade name (Krestin) oI a polysaccharide peptide, and PSP
(polysaccharide-bound peptide) extracted Irom Coriolus versicolor. A smaller percent oI
mushroom products are obtained Irom culture Iiltrates, e.g., schizophllan, a high-molecular
weight (1-3), (1-6)--D glucan, prepared Irom Schi:ophvllum commune Fr. and PSPC (a protein-
bound polysaccharide complex) Irom Tricholoma lobavense Hein. However, due to increased
quality control and year round production, mycelial products are the wave oI the Iuture.

The market value oI medicinal mushrooms and their derivative dietary supplements worldwide
was about US$1.2 billion in 1991, and about US$3.6 billion in 1994 (Chang 1996). In 1999, it
was estimated to be US$6.0 billion (Wasser et al., 2000). The market value oI Ganoderma-based
nutriceuticals alone in 1995 was estimated at US$1.6 billion (Chang and Buswell 1999). The
corresponding monetary values were also generated by another Iamous mushroom, Lentinula
edodes. Ninety nine percent oI all sales oI medicinal mushrooms and their derivatives occurred
in Asia and Europe with less than 0.1 percent in North America. The 1999 US market Ior dietary
supplements based mainly on mushrooms was estimated to be US$35 million. However, in
recent years, the North American demand is increasing between 20-40 annually, depending
upon species.

3. MUSHROOM CULTIVATION TECHNOLOGY (CONTENTS FOR A MUSHROOM
TRAINING COURSE)

3.1 Major Phases of Mushroom Cultivation

Mushroom Iarming is a complex business, which requires precision. Indeed, it is not as simple as
what some people oIten loosely stipulate. It calls Ior adherence to precise procedures. The major
practical steps/segments oI mushroom cultivation are: (a) selection oI an acceptable mushroom
species; (b) secreting a good quality Iruiting culture; (c) development oI robust spawn; (d)
preparation oI selective substrate/compost; (e) care oI mycelial (spawn) running; (I) management
oI Iruiting/ mushroom development; and (g) harvesting mushrooms careIully (Chang and Chiu,
1992, Chang 1998). II you ignore one critical step/segment, you are inviting trouble, which could
lead to a substantially reduced mushroom crop yield and mushroom marketing value.

3.1.1 Selection of Acceptable Mushroom Species/Strains

BeIore any decision to cultivate a particular mushroom is made, it is important to determine iI
that species possess organoleptic qualities acceptable to the indigenous population or to the
33
international market, iI suitable substrates Ior cultivation are plentiIul, and iI environmental
requirements Ior growth and Iruiting can be met without excessively costly systems oI
mechanical control.

3.1.2 Secreting a Good Quality Fruiting Culture

A "Iruiting culture" is deIined as a culture with the genetic capacity to Iorm Iruiting bodies under
suitable growth conditions. The stock culture which is selected should be acceptable in terms oI
yield, Ilavour, texture, Iruiting time, etc.

(1) Sources oI the cultures

(A). Tissue culture. A large healthy mushroom should be chosen either in the later button
or egg stage. It should be cleaned with 75 alcohol. The mushroom should be split in halI by
hand longitudinally and some inside tissue taken Irom the upper part oI the stipe. It should be
placed centrally on the surIace oI the medium with a sterilised needle. The quicker this is done
the better. As soon as we transIer the tissue, the test tube should be closed and dated beIore it is
returned to the incubator between 25C and 34C depending on the mushroom used. Within two
or three days some white, delicate mycelia will be produced Irom the small piece oI the tissue.
They grow upwards encircling the inner wall oI the test tube. About ten days later the mycelium
will grow rapidly and cover the surIace oI the agar medium. Then it is ready to transIer to spawn
substrate to make spawn.

(B). Spore culture. Individual spores properly collected can be transIerred singly to a test-
tube or petri dish and allowed to develop and germinate into mycelium. Some single-spore
isolates Irom homothallic mushrooms, e.g. Jolvariella volvacea (primary homothallism) or
Agaricus bisporus (secondary homothallism) can be used as Iruiting culture to make spawn.
However iI single-spore isolates are Irom heterothallic mushrooms, e.g. Lentinula edodes,
Pleurotus safor-cafu and Ganoderma lucidum, then they cannot Iorm Iruiting cultures and thus
cannot make spawn. They have to be mated with a compatible single-spore isolate. AIter mating
they Iorm a dikaryon/Iruiting culture. Then they can be used to make spawn.

(C). Pure culture Irom other laboratories. As an alternative to culturing in the laboratory,
as outlined, a test tube culture may be obtained Irom a research laboratory. The advantage oI this
is that cultures maintained in reputable culture collections are already tested Ior their production
characteristics and are guaranteed to be pure.

(D). Cultures Irom another source. Cultures also may be grown Irom spawn obtained
Irom another source. A piece oI the spawn is aseptically transIerred to agar slants. However, this
is risky because the number oI transIers that the spawn culture has undergone is rarely known. II
this procedure is Iollowed, it may be advisable to Iirst grow the spawn into Iruiting bodies, then
make the necessary isolations Irom the Iruiting body.

(2) Culture media

34
Mushrooms grow on a variety oI culture media and on diIIerent agar Iormulas, both natural and
synthetic, depending on the organism to be cultivated and the purpose oI the cultivation.
Synthetic media are oIten expensive and time-consuming in preparation hence they are not
commonly used Ior routine purposes in mushroom laboratories.

(A). PDA (potato dextrose agar), is the simplest and the most popular medium Ior
growing mycelia oI most cultivated mushrooms. It can be purchased commercially as ready
mixed powder which can be used directly to make the medium in the laboratory, with a
concentration oI 20gm/1 litre oI distilled water.

(B). Alternatively, it can be prepared in the laboratory with the Iollowing ingredients:
Potato, diced - 200 gm (1/5kg); Dextrose - 20 gm; Powdered agar or agar bars - 20gm; Water - 1
litre. Procedure: Peeled potatoes are washed, weighed, and cut into cubes. They are boiled in a
casserole with at least one litre oI water until they become soIt (around 15 minutes). The
potatoes are removed and water is added to the broth to make exactly 1 litre. The broth is
returned to the casserole, and dextrose and the agar added. The solution is heated and stirred
occasionally until the agar is melted. The hot solution is then poured into clear Ilat bottles Iilling
to about 2.5cm Irom the bottom. When using test tubes Ior the stock cultures, they are Iilled with
at least 10ml oI the liquid agar solution. The bottles or test tubes are plugged with cottonwool.
When Petri dishes are available and are used to produce mycelial colonies, the solution can be
poured into the dishes to Iorm a layer on the bottom.

(C). A ready-made MEA (malt extract agar) powder is also available commercially. The
recommended amount oI powder (20gm) is mixed with 1 litre oI water, then melted and
sterilised. One percent peptone or 0.5 yeast may be added Ior Iaster mycelial growth Ior both
PDA and MEA.

3.1.3 Development of Robust Spawn

A medium through which the mycelium oI a Iruiting culture has grown and which serves as the
inoculum oI "seed" Ior the substrate in mushroom cultivation is called the "mushroom spawn".
Failure to achieve a satisIactory harvest may oIten be traced to unsatisIactory spawn used.
Consideration must also be given to the nature oI the spawn substrate since this inIluences
rapidity oI growth in the spawn medium as well as the rate oI mycelial growth and Iilling oI the
beds Iollowing inoculation.

(1) Mushroom Spawn Preparation

(A). DeIinitions oI spawn and spawning. The word 'spawn is derived Irom an old
French verb, espandre, meaning to spread out or expand, which was derived Irom the Latin,
expandere, meaning to spread. Spawn is also deIined by Webster`s Dictionary as 'the mycelium
oI Iungi, especially oI mushrooms grown to be eaten, used Ior propagation. In the mushroom
industry, spawn is a substrate into which a mushroom mycelium has impregnated and developed,
and which will be used as a seed in propagation Ior mushroom production. In addition the verb,
to spawn, is used to mean inoculation oI a substrate with mushroom spawn.

35
The simple deIinition oI spawning is the planting oI mushroom spawn in the prepared
compost/substrate. Along with advances in spawn making, the methods oI spawning have also
been continuously developed and improved, making it possible Ior the mushroom mycelium to
grow through the compost more quickly.

(B). Spawn Substrates. A number oI materials, mostly agricultural wastes, can be used to
prepare mushroom spawn. The type oI waste available varies Irom region to region. Some oI
these wastes are chopped rice straw, sawdust, water hyacinth leaves, used tea leaves, cotton
wastes and lotus seed husks. In most laboratories, cereal grains (wheat, rye or sorghum) are used
as mother spawn, and agricultural wastes as the planting spawn substrates. The mother spawn is
used to inoculate the Iinal spawn container in which the planting spawn will be produced. The
planting spawn is used to inoculate the mushroom cultivation compost/substrate Ior
Iruiting/mushroom production.

(C). Preparation oI mother spawn. Here using wheat grains as an example, they are
soaked in water Ior 2 hours or over night. Dead seeds or those that Iloat on water should be
careIully removed. Then the grains are washed again and boiled in water Ior at least 10 to 15
minutes until they expand but not quite broken. The grains are drained and allowed to cool.
Precipitated chalk (1.5 on wet basis) is added to the grains. The grains are then loosely packed
in bottles which are 2/3 Iull. These are plugged with cotton wool or covered by double-layered
aluminium Ioil. The grains are sterilised in a pressure cooker Ior about 1 hour at 121C,
alternatively, they are steamed Ior 3 to 4 hours in a large cast-iron casserole. The bottles are then
cooled prior to inoculation.

(D). Preparation oI planting spawn. Here we shall use rice straw or water hyacinth leaves
as an example Ior Pleurotus safor-cafu planting spawn. The rice straw (or water hyacinth leaves)
is chopped into pieces about 2 to 3 cm (1 inch) long, then soaked in water Ior 4 - 12 hours. The
excess water is drained oII and the straw pieces mixed with a solution oI 1 sucrose, 1.5 chalk
and 2 wheat/rice bran in water. The Iinal moisture content is to be adjusted to about 60. The
mixture is then put into glass bottles or plastic bags and sterilised Ior at least one hour at 121C
beIore being inoculated with the mother spawn.

(E). Remarks Ior spawn making. Autoclaved substrate can only be justiIied Ior spawn
production iI it is properly done. Otherwise there will be wastage oI energy and money through
contamination losses.

(F) What means properly?
Substrates such as saw dust, straw, cotton seed hulls and so on should not be wet (iI water stands
on the bottom, mycelia will not enter). Water that is held by capillary Iorces, and swollen water,
will not as readily produce vapour pressure, as does standing water. II the container oI spawn is
tightly sealed, air cannot escape, and steam cannot enter properly. Autoclaving is thus imperIect.
AIter proper sterilisation all moulds inside are killed.

Prevent entry oI moulds Irom outside by: Using very clean cotton stoppers only (which can be
used several times, iI you observe cleanliness!); leaving at least 3-4 cm Iree space between lower
surIace oI cotton stopper and the substrate; avoiding the soilage oI the container walls around the
36
stopper, and between the stopper and the substrate surIace; preventing the cotton plug Irom
getting wet during autoclaving (cover loosely with aluminium-Ioil, so that the outside oI the
container is protected all around, at least 3 cm deep); and keeping the outside oI the containers
clean, to where the aluminium-Ioil reaches ('collar oI plastic bags).

Inoculate under clean conditions in a room without air movement (close door and windows):
clean the table with a disinIectant; wash your hands thoroughly with soap, dry with a clean
towel, and disinIect them; transIer the container Irom the autoclave directly into the clean room.
Cover them with a Ireshly washed and ironed towel iI the journey Irom the autoclave to the
inoculation place is long; keep the containers during inoculation in a position that minimises
contamination risks; use pure culture spawn only Ior inoculation (this may be Irom another
spawn batch); and cover the opening aIter inoculation with aluminium-Ioil and press around the
neck oI the container. Place the spawn in the Iirst days under optimal growth conditions. Spawn
which is not needed Ior inoculation can be used Ior Iruit body production under suitable
conditions. NOTE: Actually, one test tube oI pure culture ('starter) can prepare 1,200
commercial planting spawns through the steps oI stock culture, then the mother grain spawn.
This process is called the Multiplication oI cultures.

(2) Mushroom Spawn Handling

(A). Maintenance oI spawn quality. Mushroom spawn, whether prepared as a Iamily
home project or on industrial scale using modern equipment, should be in excellent condition
when delivered to growers. Spawn oI most mushrooms can be reIrigerated, but it should be
warmed to normal room temperature beIore it is used as an inoculum or as planting spawn.
Vigorous growth oI the planting spawn is a prerequisite to good growth and yield. II the spawn is
not vigorous, the mushroom mycelium will be overgrown by competitor organisms. II it is
vigorous it will overcome many oI the competitive organisms and produce more mushrooms.
When purchasing spawn, ask the spawn maker how long the spawn can be kept beIore planting.
Old spawn is not acceptable because its vigour may have decreased. Buyers or users should
know the 'expiration date oI the spawn.

(B). Spawn Quantities. The quantity oI the spawn used does not directly aIIect yield.
However, the use oI more spawn may reduce the eIIect oI competitive organisms present in the
planting substrates. The greater the amount oI spawn used, the Iaster it will colonise the
substrate. As a result, the growth oI competitors is hindered, and yield will be regular and not
aIIected by this competition. 2-4 oI spawn is suggested to be inoculated into the spawning
substrate. Once the container is opened, spawn should be used in its entirety. Unused and opened
bottles or bags oI spawn, however, can be kept in the reIrigerator Ior 2 to 6 days as long as they
are not contaminated (i.e., no unwanted Iungi are growing on the surIace) during storage.

(C). Notes on commercial spawn supplies. When growing mushrooms on a small scale, it
is not necessary to prepare your own spawn. Commercial suppliers oI spawn who provide
material to small growers are usually available. Spawn should be ordered ahead oI time so that it
will be oI the right age. Contaminated, old or no-growth spawn should never be sold to growers.
Spawn makers should maintain a testing Iacility where they can test each batch oI spawn Ior
production characteristics. Sales personnel should visit growers using their spawn so that they
37
can observe problems at Iirst hand. Problems related to spawn production must be corrected
quickly.

3.1.4 Preparation of Selective Substrate/Compost

While a sterile substrate Iree Irom all competitive micro-organisms is the ideal medium Ior
cultivating edible mushrooms, systems involving such strict hygiene are generally too costly and
impractical to operate on a large scale. Substrates Ior cultivating edible mushrooms normally
require varying degrees oI pre-treatment in order to promote growth oI the mushroom mycelium
to the practical exclusion oI other micro-organisms. The substrate must be rich in essential
nutrients in Iorms which are readily available to the mushroom, and be Iree oI toxic substances
which inhibit growth oI the spawn. Moisture content, pH and good gaseous exchange between
the substrate and the surrounding environment are important physical Iactors to consider.

(1). Mushroom substrate. Mushroom substrate may be simply deIined as a kind oI lignocellulosic
material which supports the growth, development, and Iruiting oI mushroom mycelium. The
process oI preparation oI substrate is broadly termed 'composting. The Iinal product oI
'composting is called the 'compost or prepared substrate. The process Ior preparation oI
substrates has been the subject oI much scientiIic and practical interest over the past two
decades. It should be noted that diIIerent types oI mushrooms require diIIerent types or
substrate/compost. Agaricus bisporus grows on Iermented compost which is traditionally
developed Irom wheat straw mixed with horse manure, and it requires higher nitrogen content.
Wheat straw contains about 0.62 nitrogen, and horse manure contains about 1.5 to 1.8
nitrogen. The optimum C:N (carbon:nitrogen) ratio Ior the mushroom is about 17 to 1 Ior
mycelial running. Jolvariella volvacea, the paddy straw mushroom, and Stropharia rugoso-
annulata are grown on almost raw or less composted plant residues, such as rice straw and cotton
waste materials. These cellulosic plant materials contain diIIerent amounts oI nitrogen: rice straw
0.58, cotton waste 0.65 to 1, and banana leaves 1.71. It should be understood that J.
volvacea is capable oI growing on plant material with low nitrogen content. The optimum C:N
ratio is about 75 to 80:1, but C:N ratios Irom 32 to 150:1 are almost as eIIective. Lentinula
edodes and Pleurotus spp. are Iungi that can grow on wood. In addition to being distinguished by
its high lignin content, wood can also be distinguished Irom other plant materials by its very low
nitrogen content. Woody tissues contain 0.03 to 1.0 nitrogen as compared to 0.85 to 1.71 in
herbaceous residues. The C:N ratio in most woody tissues is in the order oI 350 to 500:1. Wood-
inhabiting mushrooms are unique in that they can grow in such substrates. This suggests that
these mushrooms can metabolise large amounts oI carbohydrates, including lignin, in the
presence oI a very small amount oI nitrogen.

(2). Composting. Some early composting investigators had a good appreciation oI the biological
nature oI the composting process. These early researchers had some appreciation oI the role oI
various ecological Iactors such as temperature, oxygen, moisture, pH and nutrition in
determining the general activity and population dynamics that occur during composting. As a
working hypothesis, it may be assumed that composting conditions which produce a Iavourable
medium Ior the development oI mushrooms probably do so because they encourage the
development oI microbial population that is best able to pave the way Ior the subsequent growth
and IructiIication oI mushrooms. Such a hypothesis must take Iull cognisance oI the eIIect oI the
38
staling products oI diIIerent groups oI organisms on the mushroom development as well as the
actions oI these organisms in producing changes in the manure Iavourable to the nutrition oI
mushrooms under competitive conditions.

The substrate leIt aIter the mushrooms have been harvested is known as spent compost. This is
present in large amounts, and raises the question oI what can be done with it. It is certainly not
desirable to leave it as a possible source oI pollution. It is known that there still remains in the
spent compost a considerable amount oI lignocellulosic material in addition to the mushroom
mycelia and also other products Iormed by the metabolic activities oI the mycelium. Thus, the
spent compost should be capable oI supporting Iurther biological activities, e.g., the growth oI
another species oI edible mushroom; use as Iodder Ior livestock; as a soil conditioner and
Iertiliser; and also in bioremediation.

3.1.5 Care of Mycelial (Spawn) Running

Following composting, the substrate is placed in beds where it is generally pasteurized by steam
to kill oII potential competitive microorganisms. AIter the compost has cooled, the spawn may
be broadcast over the bed surIace and then pressed down Iirmly against the substrate to ensure
good contact, or inserted 2 to 2.5 cm deep into the substrate. Spawn running is the phase during
which mycelium grows Irom the spawn and permeates into the substrate. Good mycelial growth
is essential Ior mushroom production.

3.1.6 Management of Fruiting/Mushroom Development

Under suitable environmental conditions, which may diIIer Irom those adopted Ior spawn
running, primordial Iormation occurs and then Iollowed by the production oI Iruiting bodies.
The appearance oI mushrooms normally occurs in rhythmic cycles called "Ilushes".

3.1.7 Harvesting Mushrooms Carefully

Harvesting is carried out at diIIerent maturation stages depending upon the species and upon
consumer preIerences and market value.

3.2 Mushroom Diseases

The mushroom, like any other cultivated crop, is subject to attack by pathogens and pests. The
mushroom diseases can be caused by both Iungi and bacteria. There are Iour important Iungal
diseases oI the cultivated mushrooms, particularly reIerring to Agaricus bisporus and these are:

(1) Dry Bubble caused by Jerticillium fungicola. At vary early stages on inIection, brown spots
can be seen on mushroom caps. This stage oI the disease can be conIused with the brown blotch
disease caused by a bacterium. However, in the case oI dry bubble, iI the inIected mushrooms
continue to be incubated in a humid chamber, greyish white mycelium oI the pathogen will
develop Irom the brown spots on the mushroom caps. CONTROL: The environment conditions
most conducive to the growth oI Jerticillium - high relative humidity and high relative
temperature. ThereIore, an eIIective ecological control would be to reduce the relative humidity
39
and temperature in the growing rooms. A reduction in relative humidity Irom 90-95 to 80-85
and in temperature to 14
o
C can result in a reduction in the incidence oI the disease. Control oI
the disease depends also on Iarm hygiene consisting oI prompt disposal oI spent compost and
debris and disinIection oI aIIected bed areas. II soil is used as a casing material, it should be
pasteurized at 60
o
C Ior halI an hour with aerated steam.

(2) Wet Bubble caused by Mvcogone permiciosa. InIection oI mushroom Iruit bodies results in
malIormed stalk and cap. The surIace oI diseased mushrooms is covered by a white Ielt oI
mycelium oI the pathogen. The undiIIerentiated tissues oI the cap and stalk become necrotic with
a wet soIt rot dripping a brown liquid. The name 'wet bubble is derived Irom the Ioul-smelling,
mis-shapen wet mushroom tissues. CONTROL: DisinIection oI casing material or steam-air
pasteurization oI soil, iI soil is used as a casing material, can control the disease. It is necessary
to protect the developing mushroom Iruiting bodies throughout the period oI cropping. It is
essential that management gives Iull attention to hygiene such as disinIecting the work area,
removing all spent materials, disinIecting all harvesting tools and carrying out aIter-crop
sterilization using steam at 70
o
C Ior 12 hours.

(3) Mildew caused by Cladobotrvm sp. This disease is characterized by a cotton wool type oI
growth oI the pathogen on the mushroom Iruiting bodies and spreads over the casing layer to
neighbouring mushrooms. The greyish-white mycelium oI the pathogen can envelop the
mushroom Iruiting body completely. InIected mushrooms die with a soIt wet rot. CONTROL:
DisinIection oI the casing material and strict hygiene can control the disease.

(4) Aphanocladium Disease caused by Aphanocladium album. InIected mushrooms develop
brown spots which, under conditions oI high relative humidity, show greyish-white mycelium oI
the pathogen. The gill tissues oI the mushroom Iruiting body are commonly inIected by the
pathogen. CONTROL: DisinIection oI the casing material appears to control the disease.
Management oI the crop environment similar to that oI the Dry and Wet Bubble diseases can be
eIIective in controlling the disease.

In addition, there are three important bacterial diseases oI the cultivated mushrooms, particularly
reIerring to Agaricus bisporus:

(1) Brown Blotch caused by Pseudomonas tolaasii. The disease causes brown, slightly sunken
blotches on the developing mushroom Iruiting body. It aIIects only the surIace layers oI the
mushroom and, in severe cases, brown streaks develop on the stalk. The blotches on the cap can,
sometimes, be yellowish to pale brown in colour. CONTROL: The disease can be controlled by
manipulating the crop environment. Since the pathogen can spread by splashing water, the
surIaces oI mushrooms should be maintained relatively dry. II the surIaces oI mushrooms dry
within an hour or two aIter becoming wet, no inIection occurs. The drying can take place in an
atmosphere oI high relative humidity as long as there is no Iluctuation in temperature and there is
a circulation oI air over the mushroom beds. The temperature oI the air coming into the growing
room should, thereIore, be closely controlled.

(2) Mummy Disease caused by Pseudomonas sp. In diseased mushrooms the stalk is bent and the
cap tilted. OIten there is a dense growth oI mycelium around the base oI the stalk on the surIace
40
oI the casing layer. InIected mushrooms remain intact Ior a relatively long period aIter which
they are invaded by secondary bacteria. Mushrooms oIten Iail to mature and remain in the
'button stage with unopened veil. CONTROL: Strict hygiene is the best means oI controlling
the disease. DisinIecting the casing layer can help to reduce the inIection.

(3) Drippy Gill caused by Pseudomonas agarici. The pathogen damages the gill tissues oI
mature mushroom Iruiting bodies. Damage is restricted to the gills. Dark brown, round spots
appear on the side and bottom edges oI the gills. At the centre oI each spot, a creamy grey
droplet may show on the gill surIace. II the inIection is severe, bacterial droplets join to Iorm
long streaks oI slime which may lead to distortion and collapse oI the gills. CONTROL:
DisinIection oI casing soil can control the disease.

3.3 Post-Harvest Handling

Like all Iruits and vegetables, mushrooms are perishable, and aIter harvest they oIten change in
ways that make them unacceptable Ior human consumption. The most readily observable oI these
changes include wilting, ripening, browning, liqueIaction, loss oI moisture, and loss oI texture,
aroma and Ilavour. To ensure that mushrooms are acceptable and nutritious to the consumer at
the time oI purchase, it is necessary to delay or prevent senescence. Expansion oI the pileus by
growth oI gills and elongation oI the stipe post harvest is supported by increased cell wall chitin
and protein. Chitin synthase can be activated by proteinases. It has shown that during ageing,
there is a major redistribution oI dry weight between tissues with gill dry weight increasing
during storage while that oI the pileus and stipe diminishing. The rate oI cap opening depends on
the stipe length (the longer the stipe, the greater the expansion), indicating that the stipe is acting
as a major nutrient source Ior the expanding gill tissue.

Recently, the biochemistry and genetic regulation in mushrooms post harvest has been studied by
identiIying genes with higher transcript levels aIter harvest. Technologies such as cooling and
modiIied atmosphere packaging can be use to delay the rate oI senescence, while preservative
technologies such as canning, drying, pickling, and Ireezing and -irradiation arrest biological
Iunction to prevent senescence. Depending on the species, the shelI liIe oI mushrooms may vary
Irom one day to two weeks. Fresh mushrooms are best-stored unwashed in brown paper bags in
the reIrigerator, preIerably on the lowest shelI. Bags should be available Irom your mushroom
retailer; otherwise, a paper lunch bag is Iine. Generally, it is important that Iresh mushrooms are
packaged in materials that allow them to breathe, so they do not sweat` and become slimy. At
the same time, the material should ensure mushrooms do not dry out too much.

3.4 Cultivation of Several Selected Mushrooms

The cultivation oI edible mushrooms can be divided into two major stages. The Iirst stage
(vegetative) involves the preparation oI the Iruiting culture, stock culture, mother spawn and
planting spawn, while the second stage (reproductive) entails the preparation oI the growth
substrates Ior mushroom cultivation. Cultivation conditions Ior a Iew selected mushroom
species are brieIly described in the Iollowing sections. Examples oI Iormulas in the Iollowing
sections are Ior reIerence only. They should be modiIied according to the local available
materials and climatic conditions.
41

3.4.1 Agaricus bisporus (Champignon, Button mushroom) as shown in Figure 7


Figure7. Agaricus mushrooms grown on horse manure compost.

Agaricus bisporus is variously known as the white mushroom, button mushroom,
champignon, or simply the common cultivated mushroom. In Western countries this
mushroom has developed over the past 60 years Irom beginning as a risky venture to a largely
predictable and controllable industrial process, particularly in Great Britain and the Netherlands.
In no small measure this remarkable achievement in modern mushroom industrial development
may be attributed to contributions resulting Irom the vigorous research activities conducted at
mushroom research laboratories, centres and stations.

The basic principles and practical methods oI cultivation oI this mushroom have been very well
established through repetitive practical experiences during the last 60 years. It is not intended to
summarize those in this manual, instead readers are reIerred to San Antonio (1975), Chang and
Hayes (1978), Van Griensven (1988), Quimio et al. 91990) and Kaul and Dhar (2007). However,
the compost and composting are reviewed here because they play an important role in the
cultivation oI this very popular edible mushroom (this discussion is an extension oI that in
Section 3.1.4 and more speciIic Ior this mushroom Iurther reading: Hayes 1975; Nair 1991;
Nair 1994).

Generally, composting reIers to the piling up oI substrates Ior a certain period oI time and the
changes due to the activities oI various micro-organisms, which result in the composted substrate
being chemically and physically diIIerent Irom the starting material. This is sometimes reIerred
to as a solid state Iermentation. Two types oI composting are commonly described. One type
involves the decomposition oI heaps oI organic wastes and the subsequent application oI the
residue to the soil. The aim oI this type oI composting is to reduce, in a sanitary manner, both the
volume and the C:N ratio oI the organic waste so that is it suitable Ior manuring the soil to
improve the growth oI plant crops. When given directly to the soil without composting, organic
42
waste with a high C:N ratio (such as straw) can give rise to a temporary nitrogen deIiciency
which will then result in a reduction in yield oI the plant crop.

The second type oI composting is also a process oI microbial Iermentation, but in this case the
substrate is used Ior the cultivation oI edible mushrooms. Through composting, a mixture oI rich
organic materials is converted into a stable medium which is selective Ior the growth oI a
particular mushroom but is not suitable, or is less Iavourable, Ior the growth oI competing micro-
organisms. The competitors exist in uncomposted materials, and oIten in partially composted
materials, but they are Iar less active in well-composed mushroom substrates. Actually, this type
oI composting is derived mainly Irom the Agaricus mushroom-growing industry, in which a
composting technique which renders wheat straw with horse manure speciIic Ior the growth oI
the Agaricus mushroom has been developed. It should be noted that the treatment oI substrates
Ior growing other mushrooms can be regarded as 'composting, but the procedures Iollowed in
composting and the nature oI the product can be quite diIIerent. This is because the starting
materials and the lengths oI time accompanying the various changes in the substrates vary Irom
mushroom to mushroom.

As stated above, the role oI composting is the production oI a selective substrate that will
preIerentially support the growth oI the mycelium oI the mushroom. The basis oI this selectivity,
however, cannot be attributed to one Iactor or one aspect oI the entire system. The physical,
chemical and biological aspects oI composting are Iundamentally interrelated, but can be
artiIicially separated Ior the convenience oI investigation and discussion.

Mushroom growers like to practise the liberal use oI the sense oI sight, smell and touch to
evaluate the progression oI the composting process and the quality oI the Iinal product. The gross
characteristics oI compost, usually reIerred to as 'structure, result Irom a number oI complex
physical, chemical and microbial processes.

The overall goal oI composting is to produce selective nutrient media Ior the growth oI the
mushroom. These selected nutrient-rich substrates should support a high yield oI good quality
mushrooms. The general aspects oI the achievement oI composting are summarised as involving:

1) straw soItening and other structural changes;
2) modiIication oI plant materials so that nutrients are made available to mushroom growth and
development;
3) building up oI an appropriate biomass and a variety oI microbial products( some oI these can
serve as nutrient sources Ior the mushroom);
4) establishment oI selectivity, i.e. the compost promotes the growth oI the mushroom over
competitor organisms;
5) modiIication oI compost structure so that it holds more water; and
6) building up oI compost moisture content to serve as a water reservoir Ior the mushroom crop.


Composting is prepared in accordance with well-documented commercial procedures (van
Griensvan, 1988, Chang and Hayes, 1978, and Kaul and Dhar, 2007). In Phase I oI the process
(outdoor composting), locally available raw materials are arranged into piles which are
43
periodically turned and watered. The initial breakdown oI the raw ingredients by micro-
organisms takes place in Phase I. This phase is usually complete within 9-12 days, when the
materials have become pliable, dark brown in colour and capable oI holding water. There is
normally a strong smell oI ammonia. Phase II (indoor Iermentation) is pasteurisation, when
undesirable organisms are removed Irom the compost. This is carried out in a steaming room
where the air temperature is held at 60C Ior at least 4 hours. The temperature is then lowered to
50 C Ior 8 to 72 hours depending upon the nature oI the compost. CO
2
is maintained at 1.5 to
2 and the ammonia level drops below 10 PPM. Following Phase II composting, the substrate is
cooled to 30 C Ior A. bitorquis and to 25 C Ior A. bisporus Ior spawning. Production oI Phase
III or Phase IV composts Ior growing Agaricus mushrooms has been an advanced technological
development in recent years in Western countries. The production oI Phase III compost is Phase
II compost spawn run in a bulk tunnel, and ready Ior casing when delivered to the grower. II the
Phase III compost is then cased and spawn developed into casing layer beIore dispatching to the
growing unit or delivering to growers, it is named as Phase IV compost. The successes oI bulk
Phase III and Phase IV depend a lot on the quality oI Phase I and Phase II processes. Phase II on
shelves produce an average oI 4.1 crops per year. Since 1999, growers using Phase III production
enjoyed an average oI 7.1 crops per year. In recent years, Phase IV can generate 10-12 crops per
year (Dewhurst 2002, Lemmers 2003).

3.4.2 Lentinula edodes (Xiang-gu, Shiitake, Oak mushroom) as shown in Figure 8



Figure 8. Lentinula edodes grown on sawdust synthetic logs.

Lentinula edodes (Berk) Sing., (common name: black Iorest/oak mushroom; Chinese name:
Xiang-gu; Japanese name: shiitake) is the most important edible mushroom in the world Irom the
standpoint production; and it is the most popular Iungus cultivated in China, and in other Asian
countries. For a long time, this mushroom has been valued Ior its unique taste and Ilavour and as
a medicinal tonic. It can be cultivated either on wood log or on synthetic substrate logs (Quimio,
et al., 1990, Stamets, 2000, Chang and Miles, 2004).

(1). Biological nature: Lentinula edodes is a heterothallic mushroom. Its sexuality is controlled
by two mating Iactors, A and B, with multiple alleles, and thereIore, its liIe history is a tetrapolar
or biIactorial mating system (Chang and Miles, 1984). Its liIe cycle starts with the germination oI
44
basidiospores. AIter selected mating between two compatibility germinative mycelium, the
dikaryon mycelium or Iruiting culture is established. From the Iruiting culture, the stock culture,
mother spawn and commercially planting spawn can be made. When the spawn is planted on a
suitable substrate, under good climatic conditions the Iruiting bodies oI the mushroom are
developed. Then when the mature stage is reached, the spores are discharged and its liIe cycle is
completed.

Lentinula edodes is a kind oI wood rot Iungus. In nature, it grows on dead tree trunks or stumps.
In general, the wood Ior the mushroom growth consists oI crude protein 0.38, Iat 4.5, soluble
sugar 0.56, total nitrogen 0.148, cellulose 52.7, lignin 18.09 and ash 0.56. Generally
speaking, the C:N ratio in the substrate should be in the range Irom 25 to 40: 1 in the vegetative
growth stage and Irom 40 to 73: 1 in the reproductive stage. II nitrogen source is too rich in the
reproductive phase, Iruiting bodies oI the mushroom are usually not Iormed and developed.

The optimum temperature oI spore germination is 22-26
o
C. The temperature Ior mycelial growth
ranges Irom 5-35
o
C, but the optimum temperature is 23-25
o
C. Generally speaking, Lentinula
edodes belongs to low temperature mushrooms, the initial and development temperature oI
Iruiting body Iormation is in the range oI 10-20
o
C and the optimum temperature oI IructiIication
Ior most varieties oI the mushroom is about 15
o
C. Some variety can Iruit in higher temperatures,
e.g. 20-23
o
C. These high temperature mushrooms usually grow Iaster and have a bigger and
thinner cap (pileus), thin and long stalk (stipe). Their Iruiting bodies are easily opened and
become Ilat grade mushrooms, which are considered to be low quality. The optimum pH oI the
substrate used in making the mushroom bag/log is about 5.0-5.5.

(2) Culture media and preparation: The mushroom can grow on a variety oI culture media and on
diIIerent agar Iormulations, both natural and synthetic, depending on the purpose oI the
cultivation. Synthetic media are oIten expensive and time-consuming in preparation; hence they
are not commonly used Ior routine purposes.

The potato dextrose agar, or PDA, is the simplest and the most popular medium Ior growing the
mycelium oI the mushroom. It can be prepared Iollowing the instructions in Section 3.1.2.

(3). Examples oI the diIIerent Iormulas Ior spawn substrates are described below.
Mother grain spawn: (i) Wheat/rye grain 1.5 gypsum or slaked lime. (ii) Cotton seed hull
40, sawdust 38, wheat bran 20, sugar 1 and gypsum 1. (iii) Sugar cane bagasse 40,
sawdust 38, wheat bran 20, sugar 1 and gypsum 1. Planting spawn: A number oI
materials, mostly agricultural and Iorest wastes can be used to prepare mushroom planting
spawn. Three oI them are given here as examples: (i) Sawdust 78, rice/wheat bran 16, sugar
1.5, corn Ilour 1.7, ammonium sulphate 0.3, Calcium superphosphate 0.5 and gypsum
2; (ii) Sawdust 64, wheat bran 15, spent coIIee grounds 20 and gypsum/lime 1; and
(iii) Sawdust 78, sucrose 1, wheat bran 20 and Calcium carbonate 1.

The Lentinula edodes mushroom is produced both on a cottage and a commercial scale. The
Iollowing section outlines some oI the issues associated with the methodologies Ior the diIIerent
cultivation styles.

45
(1). Cottage scale cultivation: There are many Iormulas Ior the composition oI the substrate. The
ingredients can be variable Irom place to place and country to country depending upon the raw
materials available and local climatic conditions. In general, aIter mixing the dry ingredients by
hand or with a mechanical mixer, water is added to the mixture so that the Iinal moisture content
oI the substrate is between 55 and 60, depending on the capacity oI 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. AIter the bags have been Iilled
with the substrate (1.5 to 4 kg wet weight, w/w), the end oI the bag can be closed either by
strings or plugged with cottonwool stopper.

Four Iormulas in the preparation oI the substrate Ior the cultivation oI the mushroom are given
here as reIerence. (i) Sawdust 82, wheat bran 16, gypsum 1.4, Potassium phosphate,
dibasic 0.2, and lime 0.4. (ii) Sawdust 54, spent coIIee 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 hard wood
sawdust medium to grow the mushroom. The basic steps are (i) to mix the sawdust,
supplements, and water; (ii) bag the mixture; (iii) autoclave the bags to 121
o
C and cool the bags;
(iv) inoculate and seal the bags; (v) incubate Ior 90 days to achieve Iull colonisation oI the
sawdust mixture, in other words, to allow the mycelium to be established Ior ready IructiIication;
(vi) Iruit the colonised and established sawdust logs/bags/blocks 6 times using a 21 days cycle at
16 to 18
o
C; and (vii) harvest, clip steps, grade, box, and cold store Ior Iresh market, or harvest,
dry, cut steps, grade and dry again beIore box Ior dry market.

Major equipment used in production consists oI mixer/conveyor, autoclave, gas boiler, cooling
tunnel, laminar-Ilow cabinet, bag sealer, air compressor Ior humidiIication, shelves to incubate.
Incubation can be done in two rooms and in two shipping containers. The two shipping
containers can be installed near the Iruiting rooms. Temperature during incubation is held
between 18 to 25
o
C.

Fruiting can be done in 6 rooms so that the blocks/logs can be moved as a unit. With
compartmentalization, blocks in each room can be subjected to a cycle oI humid cold, humid
heat, and dry heat.

3.4.3 Pleurotus sajor-caju (Grey oyster mushroom, Phoenix-tail mushroom, Indian oyster)
as shown in Figure 9

46


Figure 9. Pleurotus safor-cafu grown on cereal straw substrates.

Pleurotus safor-cafu (grey oyster mushroom) is comparable to the high temperature species in
the group oI Pleurotus (oyster) mushrooms, with high temperatures required Ior IructiIication.
This mushroom has a promising prospect in the 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 Ior growth oI mycelium is 10-35
o
C. The optimum
growing temperature oI the mycelium is 23-28
o
C. The optimum developmental temperature oI
the Iruiting body is 18-24
o
C. The optimum pH oI 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 oI 30-60: 1. A large circulation
oI air and reasonable light are required Ior the development oI the Iruiting bodies.

(2). Examples oI spawn substrates: (i) Wheat grain 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 coIIee grounds/spent tea leaves 20, water
hyacinth/cereal straw 20, sugar 1 and gypsum 1.

(3). Examples oI cultivation substrates: (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 purposes, this mushroom can be nurtured to grow into a tree-like shape
(Chang and Li, 1982). The cultivation method, which has been tested to be successIul, is as
Iollows: Cotton waste or rice straw mixed with water hyacinth is used as the substrate. Tear large
pieces oI cotton waste into small parts or cut the straw and water hyacinth into small segments.
Add 2 per cent (w/w) lime and mix with suIIicient water to get moisture content oI about 60-65
per cent. Pile the materials up, cover with plastic sheets and leave to stand overnight. Load the
substrate into small baskets or on shelves Ior pasteurisation or cook the substrate with boiled
water Ior 15 minutes. AIter cooling to approximately 25
o
C, mix around 2 per cent (w/w) spawn
thoroughly with the substrate and pack into columns oI 60 cm long tubes which have hard plastic
(PVC) tubing oI 100 cm (4 cm in diameter) as central support, and with plastic sheets as outside
wrapping.
47

Incubate these columns at around 24-28
o
C, preIerably in the dark. When the mycelium oI the
mushroom has ramiIied the entire column oI substrate aIter three to Iour weeks, remove the
plastic wrapping and switch on white light. Watering occasionally is needed to keep the surIace
Irom drying. In around three to Iour days white primordia start to appear over the whole surIace.
AIter another two to three days, the Pleurotus mushrooms are ready Ior harvesting. During the
cropping period watering is very important iI many Ilushes are required.

3.4.4 Jolvariella volvacea (Patty straw mushroom, Chinese mushroom) as shown in Figure
10



Figure10. DiIIerent stages oI Iruiting bodies oI the Straw mushroom (Jolvariella volvacea)
grown on cotton waste as substrate.

The edible straw mushroom, Jolvariella volvacea is a Iungus oI the tropics and subtropics and
has been traditionally cultivated in rice straw Ior many yeas in China and in South East Asian
countries. In 1971, cotton wastes were Iirst introduced as heating material Ior 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 oI straw mushroom cultivation, because the cotton-waste compost through pasteurisation
process brought the cultivation oI the mushroom into an industrial scale Iirst in Hong Kong and
then in Taiwan, Thailand, and China. Several techniques are adopted Ior the cultivation oI the
mushroom, which thrives in the temperature range oI 28-36 C and a relative humidity oI 75-
85. Detailed descriptions oI the various methods are given by Chang and Quimio (1982),
Chang and Miles (2004), Kaul and Dhar (2007) and Quimio, et al. (1990). Choice oI technology
usually depends on personal preIerence, and on the availability oI substrates and the amount oI
resources available. While the more sophisticated indoor technology is recommended Ior an
industrial-scale production oI the mushroom, most oI the other technologies are low-cost and
appropriate Ior rural area development, especially when production is established at the
community level.

3.4.5 Agaricus brasiliensis (Royal Sun Agaricus, Himematuatake) as shown in Figure 11.

48


Figure 11. DiIIerent stages oI Agaricus brasiliensis mushroom grown in straw compost with case
soil.

In recent years, Agaricus brasiliensis, Iormerly called Agaricus bla:ei 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 eIIective medicinal mushroom, particularly Ior anti-
tumour active polysaccharides.

Agaricus brasiliensis was a wild mushroom in south eastern Brazil, where it was consumed by
the people as a part oI their diet. The culture oI 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 Ior commercial cultivation (Chang and Miles, 2004).

(1) Biological nature: A. brasiliensis belongs to middle temperature mushrooms. The growth
temperature Ior mycelium ranges Irom 15 to 35
o
C and the optimum growth temperature ranges
Irom 23
o
C to 27
o
C. The temperature Ior Iruiting can be Irom 16
o
C to 30
o
C and the optimum
developmental temperature oI Iruiting bodies is 18 to 25
o
C. The ideal humidity Ior casing soil is
60-65. The preIerred air humidity in a mushroom house is 60-75 Ior mycelium growth and
70-85 Ior Iruiting body Iormation and development. The optimum pH oI the compost used in
making the mushroom bed is 6.5-6.8. The optimum pH oI the casing soil is 7.0. A good
circulation oI air is required Ior the development oI the Iruiting bodies. These conditions are
similar to those needed Ior the cultivation oI Agaricus bisporus. Under natural conditions, the
mushroom can be cultivated Ior two crops each year. Each crop can harvest three Ilushes.
According to the local climate, the Iarmer can decide the spawning time in the year in order to
have mushrooms Ior harvest within 50 days aIter spawning.

(2). Preparation oI mushroom bed (Stamets, 2000): A. brasiliensis is a kind oI mushroom
belonging to straw-dung Iungi and preIers to grow on substrate rich in cellulose. The waste/by-
productive agro-industrial materials, e.g. rice straw, wheat straw, bagasse (squeezed residue oI
sugar cane), cotton seed hull, corn stalks, sorghum stalks and even wild grasses, can be used as
the principal component oI the compost Ior cultivation oI the mushroom. It should be noted that
these materials have to be air dried Iirst and then mixed with cattle dung, poultry manure and
some chemical Iertilisers. The Iollowing Iormulas Ior making compost are Ior reIerence only.
49
They should be modiIied 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 sulphate or urea 0.5.
(iii) Rice straw 90.6, rice bran 2.4, Iowl droppings 3.6, slaked lime 1.9, superphosphate
1.2 and ammonium sulphate/urea 0.3. (iv) Bagasse 75, cottonseed hull 13, Iowl
droppings 10, superphosphate 0.5 and slaked lime 1.5.

3.4.6 Canoderma lucidum (ling Zhi, Reishi) as shown in Figure 12.


37 37


Figure 12. The Iruiting bodies oI Ganoderma lucidum as shown growing on short-wood
segments which then were buried in the soil base Ior Iruiting.

Although the medicinal value oI G. lucidum has been treasured in China Ior more than two
thousand years, the mushroom was Iound inIrequently in nature. This lack oI availability was
largely responsible Ior the mushroom being so highly cherished and expensive. During ancient
times in China, any person who picked the mushroom Irom the natural environment and
presented it to a high-ranking oIIicial was usually well-rewarded (Chang and Miles, 2004).

ArtiIicial cultivation oI this valuable mushroom was successIully achieved in the early 1970s
and, since 1980 and particularly in China, production oI G. lucidum has developed rapidly.
Currently, the methods most widely adopted Ior 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 oI natural logs and tree stumps which are inoculated
with spawn directly under natural conditions. The third alternative technique involves the use oI
sterilized short logs about 12cm in diameter and approximately 15 cm long which allow Ior good
mycelial running. This method provides Ior a short growing cycle, higher biological eIIiciency,
good quality oI Iruiting bodies, and, consequently, superior economical beneIit. However, this
production procedure is more complex and the production costs much higher, than natural log
and tree stump methods. For this production procedure, the wood logs should be prepared Irom
broad-leaI trees, preIerably Irom oak. Felling oI the trees is usually carried out during the
50
dormant period, which is aIter deIoliation in autumn and prior to the emergence oI new leaves
the Iollowing spring. The optimum moisture content oI the log is about 45-55 .The Ilow-chart
Ior the short-log cultivation method is as Iollows : selection and Ielling oI the tree---
sawing/cutting the log into short segments---transIer segments to plastic bags---sterilization---
inoculation---spawn running--- burial oI the log in soil---tending the Iruiting bodies during
development Irom the pinhead stage to maturity---harvesting oI the Iruiting bodies---drying oI
the Iruiting bodies by electrical driers--packaging. It should be noted that the prepared
logs/segments are usually buried in soil inside a greenhouse or plastic shed. The soil should
allow optimum conditions oI drainage, air permeability and water retention, but excessive
humidity should be avoided.

Examples oI cultivation substrates, using plastic bags or bottles as containers, include the
Iollowings (please note that these examples are Ior reIerence purposes only and can be modiIied
according to the strains selected and the materials available in diIIerent 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. AIter sterilisation, the plastic bags can be laid horizontally on beds or the
ground Ior Iruiting.

4. ENVIRONMENTAL IMPACT

4.1 Reducing Environmental Pollution by Bioconversion of Vast Quantities of Organic
Wastes into Mushrooms

Organic solid wastes are a kind oI biomass, which are generated annually through the activities
oI the agricultural, Iorest and Iood processing industries. They consist mainly oI three
components: cellulose, hemicellulose and lignin. The general term Ior these organic wastes is
lignocellulose.

It is common knowledge that lignocellulosic wastes are available in abundance both in the rural
and urban areas. They have insigniIicant or less commercial value and certainly no Iood value, at
least in their original Iorm. When carelessly disposed of in the surrounding environment by
dumping or burning, these wastes are bound to lead to environmental pollution and
consequently health hazards. It should be recognised that the wastes are resources out oI place
and their proper management and utilization would lead to Iurther economic growth as well.

Huge quantities oI lignocellulosic and other organic waste residues are generated annually
through the activities oI agricultural, Iorest and Iood processing industries. In 1999, more than
3,000 million tons oI cereal straws were available in the world, and about halI oI these residues
remain unused. In addition, the world produced 952 million tons oI bagasse; 6,476 thousand tons
oI coIIee pulps; 6,152 thousand tons oI coIIee wastes; 9,386 thousand oI cottonseed hulls;
14,073 thousand tons oI sunIlower seed hulls; and 325 thousand tons oI sisal wastes. Million
tons oI sawdust, wood chips, and water hyacinth are also available worldwide. All these
lignocellulosic waste residues can be used as substrate growing mushrooms; otherwise, they
51
would cause health hazards. Mushroom enzymes can break down lignin, cellulose and
hemicellulose present in these organic materials into simpler molecules, which the mushrooms
then use Ior their growth and metabolism.

Lignocellulosic compounds are complex and insoluble. They can be treated by various chemical
methods, e.g. with dilute hydrochloric acid and calcium chloride to increase the digestibility and
nutritional qualities, and even to Iorm sugars to serve as carbon sources. However, these
chemical methods are tedious and costly. Furthermore, treatments to eliminate adverse side
eIIects oI the chemicals are also very complex. In contrast, mushroom cultivation techniques
have become signiIicantly important in recent years in improving nutritional quality and
upgrading the economic value oI the solid organic wastes. Mushrooms with other Iungi are
presently only organisms that can synthesize and excrete the relevant hydrolytic and oxidative
enzymes that enable them to degrade complex organic substrates into soluble substances which
can then be absorbed by the mushrooms Ior their nutrients,

The ability oI the diIIerent mushroom species to utilize various substrates will depend on both
mushroom-and substrate-associated Iactors. For example, examination oI the lignocellulolytic
enzymes proIiles oI the three important commercially cultivated mushrooms exhibit varying
abilities to utilise diIIerent lignocellulosics as growth substrate. Lentinula edodes is cultivated on
highly ligniIied substrates such as wood or sawdust, produces two extracellular enzymes
(manganese peroxidase and laccase) which have been associated with lignin depolymerisation.
Conversely, Jolvariella volvacea preIers high cellulose- low lignin-containing substrates such as
paddy straw and cotton wastes which have relatively low lignin content, and produces a Iamily
oI cellulolytic enzymes including at least Iive endoglucanases, Iive cellobihydrolases and two -
glucosidases, but none oI the recognised lignin-degrading enzymes. Pleurotus safor-cafu is the
most adaptable oI the three species and can be grown on a wide variety oI agricultural waste
materials oI diIIering composition in terms oI polysaccharide/lignin ration, because it is able to
excrete both kinds oI cellulose- and lignin-degrading enzymes.

4.2 Recycling of Organic Wastes into Mushrooms, Biofertilizer and Biogas

The ultimate aim in the applied aspects oI any scientiIic endeavour is to integrate wherever
possible the various disciplines oI science as well as the technological processes in order that
maximum beneIits accrue Irom such eIIorts. Combined production oI mushrooms, biogas and
bioIertilizer Irom the rural and urban organic wastes should be one oI the aims oI such integrated
schemes that can eventually be put into proIitable operation. Though the conventional and
established approaches towards the production oI Iood, Iertilizer and Iuel exist, the explosive
growth oI the population vis-a-vis the rapid depletion oI conventional Iuel resources leads
mankind to look Ior alternative sources Ior Iood, Iertilizer and Iuel.

Even though man has been harvesting mushrooms as Iood Irom wild sources Irom times
immemorial, their nutritive value was not assessed and their production under controlled
conditions was not undertaken until recent decades. The lignocellulosic substrate used Ior
mushroom production and which is leIt aIter harvesting oI the mushrooms can be used as
compost Ior soil conditioning. It should be noted that this compost besides being rich in
nitrogenous material contains partly degraded lignocellulosic components, which when
52
combined with animal dung or human excreta in a biogas digest would yield not only biogas but
also a good quality organic nitrogenous Iertilizer in the Iorm oI sludge. The sludge Irom the
biogas plant as a nitrogenous Iertilizer is Iar more beneIicial than the compost Irom which it has
been derived. Part oI the biogas that is produced in the vicinity oI the mushroom house can also
be conveniently used Ior pasteurization oI the mushroom bed material and maintenance oI the
optimal temperature in the mushroom house as well.

It is thereIore suggested that an integrated approach in the production oI mushroom, bioIertilizer
and biogas should be considered as a Ieasible approach Ior the rural and urban lignocellulosic
waste utilization and disposal. This is the 'Zero Emission or Total Productivity concept. It is
said 'The earth can not produce more: Man has to do more with what the earth produces (Pauli,
1996; Chang, 2007).

4.3 Restoration of Damaged Environment by Mushroom Mycelia

Mushroom cultivation technology is Iriendly to the environment. Mushroom mycelia can
produce a group oI complex extracellular enzymes which can degrade and utilize the
lignocellulosic wastes in order to reduce pollution. It has been revealed recently that mushroom
mycelia can play a signiIicant role in the restoration oI damaged environments. Saprotrophic,
endophytic, mycorrhizal, or even parasitic Iungi/mushrooms can be used in mycorestoration,
which can be perIormed in Iour diIIerent ways: mycoIiltration (using mycelia to Iilter water),
mycoIorestry (using mycelia to restore Iorests), mycoremediation (using mycelia to eliminate
toxic waste, and mycopesticides (using mycelia to control insect pests). These methods represent
the potential to create the clean ecosystem, where no damage will be leIt aIter Iungal
implementation (Stamets 2005).

5. ECONOMIC AND SOCIAL IMPACTS

Since mushroom cultivation can be a labour-intensive agro-industrial activity, it could have great
economic and social impact by generating income and employment Ior both women and youth,
particularly in rural areas in developing countries. Using China as an example, in 1978 the total
production oI mushrooms in China was only 60,000 tonnes, which accounted Ior less than 6
percent oI total world mushroom production. In 2006, however, total production oI mushrooms
in China reached 14 million tonnes and accounted Ior over 70 per cent oI total world mushroom
production. According to recent statistics, in 2006 the value oI total mushroom production in
China was US$6billion and the export value oI mushrooms was US$1.1billion. It is estimated
that in 2007, the export value oI mushrooms is US$1.4billion and it is expected to grow to
US$1.6billion in 2008. Total employment in the mushroom industry in China was over 30
million people in 2006, with only 10 percent oI the employed being actual mushroom Iarmers,
other employment Iall within sectors such as Iood, beverage manuIacturing, trading and
management, transport, marketing, wholesaling, retailing, export etc. The mushroom industry
can also have even broader positive spill-overs, generating complementary employment in areas
such as accommodation, restaurant services etc. Further, it is interesting to note that in some
counties in China with a population oI just under 200,000 people, 60 per cent oI the population
were engaged in mushroom production and management. The local mushroom industry can also
be the main source oI revenue Ior local government.
53

Mushrooms, like all other Iungi, lack chlorophyll and are non-green organisms. They cannot
convert solar energy through the process oI photosynthesis to organic matters as green plants do,
but they can produce extensive enzymes that can degrade lignocellulosic materials Ior their own
nutrients Ior growth and Iruiting. DiIIerent mushrooms have diIIerent lignocellulolytic enzyme
proIiles (Buswell and Chang, 1994, Buswell et al., 1996a). This demonstrates the impressive
capacities oI mushrooms Ior biosynthesis`, which is diIIerent Irom photosynthesis` by green
plants. The species oI mushroom Iungi not only can convert the agricultural and Iorestry
lignocellulosic wastes through solid Iermentation technology into the high quality protein
consumed directly in the Iorm oI the mushroom Iruiting body, but also can convert Iood
processing biomass wastes, e.g., soybean wastes using submerged culture, into Iungal protein
(Buswell and Chang 1994) or 'mycomeat (Miles and Chang 1988). Soybean waste materials
(slurries) are generated in large quantities during the processing oI soybean mild and 'toIu
(bean curd), which are popular Ioods in many countries now and are, in some places, discarded
without treatment thereby constituting an environmental pollutant. In addition, mushrooms and
their mycelia can provide nutriceutical and pharmaceutical products. As outlined in above
sections, by blending the advances in basic biological knowledge with that oI practical
technology, a mushroom-related industry based on utilization oI the lignocellulosic waste
materials that are abundantly available in rural and urban areas can have positive global impacts
on long-term Iood nutrition, health, environmental conservation and regeneration, and economic
and social change. ThereIore, the signiIicant impact oI APPLIED MUSHROOM BIOLOGY on
human welIare has been named as a 'Non -green revolution (Chang 1999).

The Iollowing statements summarise the signiIicance oI mushrooms in our drive towards
alleviating poverty, enhancing human health, and arresting environmental degradation:

(1). Mushrooms can convert lignocellulosic waste materials into a wide diversity oI products,
which have multi-beneIicial eIIects to human beings, e.g., as Iood, health tonic, and medicine, as
Ieed, as Iertilisers, and Ior protecting and regenerating the environment. In addition, mushroom
cultivation can positively generate equitable economic growth. The tropical regions, particularly,
have a wet and warm climate and have an abundant supply oI agricultural wastes. These
materials are resistant to natural biological degradation because they contain mainly cellulose,
hemicellulose and lignin. Mycelia oI mushrooms can excrete enzyme complexes which can
directly attack/degrade these components oI lignocellulosic materials. ThereIore, mushrooms can
use these wastes as nutrients Ior their growth and in the process become Iood and medicine Ior
human consumption.

(2). Mushrooms are relatively Iast growing organisms. Some tropical mushrooms can be
harvested and consumed within 10 days aIter spawning. By the use oI diIIerent varieties,
mushrooms can be cultivated year round. They can be cultivated by using primitive Iarming
techniques in rural areas or by using highly industrialised technologies in the urban and peri-
urban communities.

(3). Mushroom cultivation can be labour intensive. Thus the activity can generate new jobs,
especially in tropical or less developed countries.

54
(4). While land availability is usually a limiting Iactor in most types oI primary production,
mushroom cultivation requires relatively little space. Actually they can be stacked using shelI-
like culture systems.

(5). Mushrooms have been accepted as human Iood Irom times immemorial, and can
immediately supply additional protein to human Iood. Other sophisticated and unconventional
sources oI Iood protein, such as yeast, uni-algal cultures and single-cell proteins have relatively
more complicated requirements, and need to be processed beIore they can be consumed.

(6). Edible mushrooms should be treated as healthy vegetables. AIter improving the cultivation
techniques, they should be cultivated as widely and as cheaply as other common vegetables,
which will thus be beneIicial to the general public.

(7). In view oI their pleasing Ilavour, their high protein, and tonic and medicinal values,
mushrooms no doubt represent one oI the world`s greatest untapped resources oI nutritious and
palatable Iood Ior the Iuture.


6. CONCLUDING REMARKS

From what has been presented in the preceding pages, the Iollowing summary can serve as
concluding remarks:

---Mushrooms can serve as Iood, as tonic, and as medicine. A regular intake oI
mushrooms can make you healthier, Iitter, and happier. They can make you live longer, and
always look younger.

---Mushrooms are biota characterized by wonder. They rise up Irom lignocellulosic
wastes, yet they become so bountiIul and nourishing.

---Mushrooms are environmentally very Iriendly. They biosynthesise their own Iood
Irom agricultural crop residues, which would otherwise cause health hazards. And their spent
composts/substrates can be used as animal Ieed, bioIertilizers and biogas.

---Mushrooms can serve as agents Ior promoting equitable economic growth in society.
They are a unique group oI Iungi through which we can pilot a non-green revolution in less
developed countries, and in the world at large. They demonstrate great potential Ior generating a
great socio-economic impact in human welIare, at local, national and regional levels.

ThereIore, the aims oI the discipline oI applied mushroom biology are to tackle the three basic
problems: shortage of food, diminishing quality of human health and pollution of the
environment, which human beings still Iace, and will continue to Iace, due to the continued
increase oI the world population. On the other hand, it has been observed that over 70 oI
agricultural and oI Iorest products have not been put to total productivity, and have been
discarded as waste. Applied mushroom biology not only can convert these huge lignocellulosic
biomass wastes into human Iood, but also can produce notable nutriceutical products, which
55
have many health beneIits. Another signiIicant aspect oI applied mushroom biology is using the
biota in creating a pollution-Iree and beneIicial environment. These three components oI applied
mushroom biology are closely associated with three aspects oI wellbeing Iood shortage, human
health and environmental pollution. One of the most significant benefits of mushroom
cultivation is their ability to create a pollution free and friendly environment.

7. ACKNOWLEDGEEMENTS

First oI all, I would like to take this opportunity to express my sincerest thanks to the UN-
APCAEM Ior having invited me to serve as a consultant to advise on environmental Iriendly
mushroom cultivation technology. I am grateIul also to ProIessor Chang Ping, Senior Expert oI
UN-APCAEM in Beijing, the project coordinator, who helped me enthusiastically on all aspects
during the development oI this project, and who has shown a great interest in the mushroom
project.

8. SUGGESTED FURTHER READINGS

Beelman R. B. and D. J. Royse. 2006. Selenium enrichment oI Pleurotus cornucopiae (Paulet)
Rolland and GriIola Irondosa (Dicks.:Fr.) S. F. Gray mushrooms. Intl. J. Med. Mushr. 8, 77-84.

Bennett, J. W. and L. L. Lasure. 1991. More Gene Manipulations in Fungi. Academic Press,
INC. New York, USA.

Beyer, D. 2005. Spent mushroom substrate (SMS) research in the US. AMGA J. Summer Issue
31-32.

Buswell J. A. and S. T. Chang. 1994. Biomass and extracellular hydrolytic enzyme production
by six mushroom species grown on soybean waste. Biotechnology Letters.16: 1317-1322.

Buswell J. A., Y. J. Cai and S. T. Chang. 1995. EIIect oI nutrient nitrogen and manganese on
manganese peroxidase and laccase production by Lentinula edodes. FEMS Microbiology Letters
128: 81-88.

Buswell J. A., Y. J. Cai and S. T. Chang. 1996. Ligninolytic enzyme production and secretion in
edible mushroom Iungi. In Mushroom Biology and Mushroom Production, Royse D. J. ed.,
University Park, Pan. 113-122.

Buswell J. A., Y. J. Cai, S. T. Chang, J. F. Peabody, S. Y. Fu and H. S. Yu. 1996.
Lignocellyulolytic enzyme proIiles oI edible mushroom Iungi. World J. Microb. &
Biotechnol.12: 537-542.

Cai Y. J., J. A. Buswell and S. T. Chang. 1994. Production oI cellulases and hemicellulases by
the straw mushroom, Jolvariella volvacea. Mycol. Res. 98: 1019-1024.

56
Cai Y. J., J. A. Buswell and S. T. Chang. 1998. b-Gludosidase components oI cellulolytic system
oI the edible straw mushroom, Jolvariella volvacea. Enzyme and Microbial Technology 22: 12-
129.

Cai Y. J., S. J. Chapman, J. A. and S. T. Chang. 1999. Production and distribution oI
endoglucanaase, cellobiohydrolase, and b-Glucosiase components oI the cellulolytic system oI
Jolvariella volvacea, the edible straw mushroom. Appl. and Environ. Microbiology 65:553-559.

Chang S. T. 1965. Cultivation oI the straw mushroom in S. E. China. World Crops 17: 47-49.

Chang S. T. 1974. Production oI the straw mushroom (Jolvariella volvacea) Irom cotton wastes.
The Mushroom J. 21: 348-353.

Chang S. T. 1989. Recycling oI solid wastes with emphasis on organic wastes. Green
Productivity, March Issue 23-16.

Chang S. T. 1996. Mushroom research and development equality and mutual beneIit. In
Mushroom Biology and Mushroom Products. Royse D. J., ed., University Park, Pan. 1-10.

Chang S. T. 1998. Development oI novel agroscience industries based on bioconversion
technology. In Frontiers in Biology: the Challenges oI Biodiversity. Chou C. H. and K. T. Shao,
ed., Academia Sinica, Taipei. 217-222.

Chang S. T. 1999. Global impact oI edible and medicinal mushrooms on human welIare in the
21st century: Non-green revolution. Intl. J. Med. Mush. 1:1-7.

Chang S. T. 1999. World production oI cultivated edible and medicinal mushrooms in 1997 with
emphasis on Lentinus edodes (Berk.) Sing. in China. Intl. J. Med. Mushrooms 1:291-300.

Chang, S. T. 2006a. The need Ior scientiIic validation oI culinary medicinal mushroom
products. Intl. J. Med. Mush, 8:187-195.

Chang, S. T. 2006b. The world mushroom industry: Trends and technological development. Intl,
J. Med. Mush. 8:297-314.

Chang, S. T. 2007. Mushroom cultivation using the 'ZERI principle: potential Ior application in
Brazil.Micol. Apl. Int., 19:33-34.

Chang, S. T. and J. A. Buswell. 1996. Mushroom nutriceuticals. World J. Microb. and Biotech.
12:473-476.

Chang, S. T. and J. A. Buswell. 1999. Ganoderma lucidum (Curt.:Fr.) P. Karst.
(Aphyllophoromycetidease): a mushrooming medicinal mushroom. Int. J. Med. Mush. 1: 139-
146.

57
Chang, S. T. and J. A. Buswell. 2003. Medicinal mushroomsa prominent source oI
nutriceuticals Ior the 21st century. Current Topics in Nutraceutical Res,1:257-280.

Chang, S. T. and J. A. Buswell. 2008. Development oI the world mushroom industry: Applied
mushroom biology and international mushroom organization. Intl, J. Med. Mush. (In press).

Chang, S. T. and S. W. Chiu.1992. Mushroom productionan economic measure in
maintenance oI Iood security. In: Biotechnology : Economic and social Aspects, edited by
DaSilva, E. J., C. Ratledge and A Sasson. Cambridge University Press. Pp110-141.

Chang, S. T. & W. A. Hayes( eds.) 1978. The Biology and Cultivation oI Edible Mushrooms.
Academic Press, Inc., New York & London. 819 pp.

Chang S. T. and P. G. Miles. 1982. Introduction to mushroom science. In Tropical Mushrooms-
Biological Nature and Cultivation Methods, Chang, S. T. and T. H. Quimio, the Chinese
University Press, Hong Kong. 3-10.

Chang S. T. and P. G. Miles. 1984. A new look at cultivated mushrooms. BioScience 34: 358-
362.

Chang, S. T. and P. G. Miles. 1989. Edible mushrooms and their cultivation. CRC Press, Inc.
Florida, 345p.

Chang S. T. and P. G. Miles. 1992. Mushroom biologya new discipline. The Mycologist 6:64-
65.

Chang, S. T. and P. G. Miles. 2004. Mushrooms: Cultivation, Nutritional Value, Medicinal
EIIect, and Environmental Impact (Second Edition).CRC Press.Boca Raton, 451pp.

Chang S. T. and K. E. Mshigeni. 2001. Mushrooms and Human Health: with special reIerence to
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Letters supplement 28:S59.


9. ANNEX: RECOMMENDATIONS FOR FOLLOW-UP ACTIVITIES

Cultivated mushrooms have now become very popular all over the world. This is because
mushrooms can adapt to a wide range oI climatic conditions, and also because they have
immense nutritional and medicinal values.

The production oI mushrooms in the Asian and PaciIic area accounted Ior over 75 oI the total
world production in 2005. Those amounts were mainly contributed by the three north-eastern
countries, China, South Korea and Japan. China`s success in the development oI its mushroom
industry during the past three decades should serve as an example on what is also possible Ior
other developing countries.

9.1 Problems for Cultivation of Mushrooms in Some Countries in the Region

In practical application, the cultivation oI Agaricus mushrooms in developed countries has
become a high technology industry, and in Iundamental research, it has become a branch oI
science which is derived Irom the disciplines oI microbiology, Iermentation and environmental
engineering. In the last two decades, millions oI dollars have been spent in western countries to
develop the industry. The mushroom Iarms are usually Iurnished with heavy equipment. The
support Irom industry and governments Ior Agaricus mushroom research is equally impressive in
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some developed countries such as the USA (Mushroom Research Centre at the Pennsylvania
State University), UK (Mushroom Research Section at Horticultural Research International).

In Asia, there are also national and local government support mushroom research centres,
particularly in China, Japan, South Korea and India.

However, in other Asian and PaciIic countries, there appears to be a lack oI such inIrastructure
Ior the mushroom industry. There are several reasons/ problems, which may be directly or
indirectly related to the slow development oI mushroom cultivation in tropical regions:

(1). Social concept. Mushrooms are usually eaten Ior their culinary properties, providing a
Ilavouring and garnish Ior other Ioods. They are cultivated with special technique and usually
consumed by the rich people because the price oI mushroom is usually much higher than that oI
the most common vegetables. This may give one the impression that mushrooms constitute a
luxury Iood and that their promotion would only beneIit relatively rich people. Actually,
mushrooms are rich in protein and contain several vitamins and mineral salts and should thus be
considered as high protein vegetables to enrich all human diets.

(2). Lack oI government and industrial support Ior research. Research on tropical mushrooms is
relatively inexpensive (requiring neither huge research establishments nor massive, highly
complex equipment). As Iar as I know, no government supported mushroom research centre or
institute has been established in tropical regions.

(3). Inadequate interest on the part oI academic scientists in the Iundamental biological studies oI
edible mushrooms. This contributes to the delay in the massive production oI edible mushrooms
in the tropical regions. ThereIore, the knowledge on the biological nature oI mushrooms
cultivated in tropical regions is very meagre indeed. Without such basic knowledge, the
development oI any mushroom industry is rather diIIicult. Since the cultivation oI mushrooms,
by its nature, requires very strong regional and local adaptation, the greatest needs should be the
creation oI national and regional laboratories /centres oI Iundamental biological and applied
technical research on edible mushrooms in tropical/ subtropical countries, and also the
development oI bright, young, and highly motivated people who will be attracted work in these
laboratories / centres.

(4).Those three problems mentioned above are the most basic and important problems. There are
several other problems, such as shortage oI technical expertise, lack oI appropriate equipment,
inadequate regional cooperation, lack oI organised marketing strategies, etc that would require
particular attention.

(5). Further, due to the prevailing hot and humid climate, mushroom cultures are more easily
contaminated; the shelI liIe oI the mushrooms is diIIicult to be prolonged; and the preparation
and preservation oI the mushroom pure culture spawn is also a common problem to mushroom
growers, since they do not have basic knowledge in microbiology.

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There should be a concerted eIIort to lobby the governments and international/national agents,
reminding them that research and development oI mushroom industry is not a luxury but a
national necessity Ior human welIare.

9.2 The Strategies for Developing Mushroom Industries in the Asian and Pacific Region

(1). At the beginning, the strategy is not to use highly mechanized technologies as in the large
mushroom Iarms in industrialized countries, but to promote cottage style enterprise Ior the rural
poor in thousands oI small mushrooms sheds, constructed using locally available materials ( like
those used in China at the beginning oI its mushroom industry).
(2). Then more towards gradual introduction and Iamiliarization oI the art oI large scale
commercial cultivation techniques. This was the path China Iollowed Irom its humble
beginnings to its current status as the world`s leading mushroom production powerhouse.
(3). Select appropriate target strains oI diIIerent mushrooms grown on seasonal basis so that an
attempt could be made to obtain yields all year-round.
(4). Make use oI existing lignocellulosic residues and waste Irom agricultural activities and agro-
industries.
(5). Create employment opportunities, particularly Ior women and the youth in rural areas, and
control/reduce pollution.
(6). Emphasize quick-investment-return mushrooms, and select relatively Iast growing species
that can be harvested within 3 to 4 weeks aIter spawning, thus generating immediate beneIits.
(7). Promote mushrooms species demonstrated to generate potent nutriceuticals with superior
immune-enhancing attributes: species whose natural products include unique bioactive
compounds that can make people healthier and Iitter.

The Iollowing ideas also need to be emphasized. Although science and Iarming practice have led
to the development oI some universal or general concepts concerning mushroom cultivation, the
diverse biological nature oI the process (in which large numbers oI mushroom species and
natural organic substrates are involved) also means that a wide spectrum oI variations in Iarming
methods must be employed. Thus the transIer oI mushroom cultivation techniques Irom one
region or country to another cannot be treated in the same way as the transIer oI non-biological
industrial technology, such as that oI a complete complex oI Iactory equipment Ior textile or
chemical Iertilizer industries. Since the cultivation oI mushrooms deals with living organisms,
one should consider, not only the unique attributes oI the mushroom itselI, and oI the various
micro-organisms growing with it (including both the harmIul and beneIicial ones), but also the
biochemical nature oI the local available substrates. ThereIore, the speciIic methods must be
tailored in accordance with the prevailing unique natural resources, heritage, local climate, and
socio-economic conditions oI the Iarming community. All these consideration call Ior a critical
mass oI well trained mushroom scientists. Thus the training activities oI the UN-APACEM
Project oI Asian member countries need to be Iurther supported.

9.3 Appropriate Training for Regional Mushroom Scientists and Mushroom Farmers

(1) A one-week mushroom training workshop can be held Iirst at a convenient time covering the
topics in Section 2. The number oI participants can be around 18-24. Two to three can be
nominated Irom each oI the selected regional members, and the host country can have Iour. They
64
should have a general knowledge oI microbiology/ plant pathology/and mushroom cultivation. It
is hoped that by the end oI the Workshop, the selected participants would be equipped with new
knowledge and basic skills that will enable them to do Iurther research, training and Iarming
mushrooms, and eventually to generate robust and high quality edible and medicinal products
Irom the local biota.

(A). Content oI the Workshop: The Training Workshop is designed to cover the basic
science and technologies involved in the production oI mushrooms using agricultural and other
organic wastes, as well as other categories oI biomass (e.g., water hyacinth, which many
communities oIten conceive as environmental tragedies).

(B) Methodology: The training Workshop would involve Lecture Sessions in the
morning, and Discussion and Question Sections in the AIternoon.

(C) Resource Person: The Key Resource Person Ior the Training Workshop can be a
world known mushroom scientist.

(2). Then there should be a two- or three -week mushroom training course, which provides more
detailed and practical lessons on cultivation technology, similar to the topics in Section 3.

(A) Contents: Establishment oI a mushroom spawn Iacility;
Appropriate training Ior national scientists;
IdentiIication oI local mushrooms Ior Iarming development; and
Construction oI model mushroom Iarm Ior demonstration and training purposes.
Establishment oI a mushroom spawn Iacility: An old Chinese saying stipulates that iI
you wish to get the work done well, you must Iirst have proper Iacilities or the correct
tools. The isolation oI pure cultures oI mushrooms and the preparation oI mushroom
spawns are microbial manipulations. ThereIore, mushroom spawn workers need not
only the knowledge and appropriate techniques in microbiology, but also require a
suitable working place (laboratory), where proper equipment and Iacilities should be
provided, in order to achieve the production oI good quality spawns.

(B). Methodology: In addition to the Lecture and Discussion Sessions, there should be
Laboratory Exercise Sessions.

(C). Resource Person: At least, two resource persons should be invited.

9.4 Consideration on the Formation of a Regional Network on Mushroom Development

It may be considered in due course that the establishment oI a Network as an integral part oI the
sustainable development oI edible/medicinal mushroom industry aims to improve the capability,
productivity and business development oI the mushroom industry in the Region. The
justiIications oI this suggestion are in responding to diIIerent needs oI member countries in the
development oI mushroom industries, particularly in capacity building, diIIerent socio-economic
and environmental aspects, exchange oI inIormation on the development oI edible and medicinal
mushroom industry; emerging need to share diIIerences and similarities in experiences and
65
knowledge, and program and policy regarding the development oI mushroom industry within the
regional countries; growing demand in solving multilateral constraints on marketable products
and marketing oI edible and medicinal mushrooms in global market; and developing strategies to
promote the consumption oI edible and medicinal mushrooms.