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Praveen Gehlot · Joginder Singh Editors
This Springer imprint is published by the registered company Springer Nature Singapore Pte Ltd.
The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721,
Singapore
Contents
v
vi Contents
Dr. Praveen Gehlot is Associate Professor, Department of Botany, Jai Narain Vyas
University, Jodhpur, India. He did his Ph.D. from JNV University Jodhpur, under
the able guidance of Prof. D.K. Purohit. He has about 18 years of teaching and
research experience. He has been an awardee of Young Scientist under FAST Track
Scheme by Department of Science and Technology, New Delhi, and later worked as
a Pool-Officer of Council of Scientific and Industrial Research, New Delhi. Earlier,
he taught at Budha Institute of Technology and Science Research College, Jodhpur,
and Government Bangor PG College, Pali, Rajasthan, and subsequently served as
an Associate Professor and Head, Department of Microbiology, Maharaja Ganga
Singh University, Bikaner, Rajasthan. Dr. Gehlot has successfully completed three
major research projects sponsored by UGC, DST and CSIR. He has published more
than 75 research papers, book chapters and review articles in journals of interna-
tional and national repute and has edited three books entitled Pyrenomycetes Fungi,
Arbuscular Mycorrhizal Fungi and Microbes: In Action published by AgroBios,
India. He attended several international and national seminars, symposia, confer-
ences and chaired technical sessions. He is an active member of various scientific
societies, including Mycological Society of India, Society of Mycology and Plant
Pathology, India, Association of Microbiologists of India, and Indian Science
Congress Association.
ix
x About the Editors
1.1 Introduction
Mushrooms are saprophytic macrofungi. Unlike plants they lack chlorophyll and
draw nutrition from dead and decaying organic materials. The mycelium obtains
nutrition by penetrating substrate and at maturity or under unfavourable conditions
forms fruiting bodies of various shapes, sizes and colour. Generally, mushroom fruit
body consists of pileus and stipe, and a few mushrooms have veil or annulus, cup or
volva and perform vital functions during the life cycle of the fungus. The pileus of
the fruit body produces profuse spores of microscopic size upon maturity. These
spores are dispersed and fall in soil/substrate. Under favourable conditions of mois-
ture and temperature, these spores germinates to produces mycelia on suitable sub-
strate. This mycelium continues to grow, mate by several means and undergo
fructification to complete its life cycle. Due to saprophytic nutritional habit,
mushrooms usually grow in shady moist places on dead organic matter, decaying
plant remains, wood logs, etc. Therefore, mushrooms grow in ample in forests,
fields and meadows. However, some mushrooms are not edible and can be highly
poisonous, and commonly referred to as ‘toadstools’ are objects of fear and distrust
and have led to certain amount of inhibition towards mushroom eating. Mushroom
cultivation being an indoor activity is highly remunerative as it requires low invest-
ment, relatively small space than any other agricultural activity and has simple cul-
tivation technology.
Out of 10,000 macrofungi, only 2000 have been reported growing from different
parts of the globe. Since India has rich floral diversity than any other country of its
size, it is expected to have great fungal wealth and species diversity (Watling and
Gregory 1980). Indian mycologists especially working on fleshy fungi have either
described or reported several species from different parts of the country without
mentioning their edibility. This was probably due to their major interest in survey
and taxonomic studies of the fleshy fungi rather than their economic importance. All
the reported fungi are not edible (Verma et al. 2003).
Purkayastha and Chandra (1976) published a book entitled Indian Edible
Mushrooms in which 122 edible species have been recorded, of which 105 species
are briefly described. In the 1970s two national symposia on survey and cultivation
of macrofungi were organized in Srinagar (Atal et al. 1978; Kaul and Kapoor 1987).
Manjula (1983) enlisted 115 genera and 538 species of agaricoid and boletoid fungi
from India and Nepal. Purkayastha and Chandra (1985) published a manual of
Indian Edible Mushrooms and described more than 430 edible fungi along with
their characters and distribution in India. Bhawani Devi and Nair updated the list of
Indian Agaricales in 1987. Dhar and Singh (1989) presented an account of biodiver-
sity in Indian mushrooms during FAO workshop on Mushroom Biology and
Mushroom Products held in Sydney.
This chapter deals with cultivation, conservation and medicinal significance of
macrofungi to underline their role in sustainable development.
1.2 Cultivation
Mushroom cultures are required for variety of purposes such as preparation of qual-
ity spawn, conservation of genetic resources and maintenance of true to the type
mushroom strains. The mushroom pure cultures are usually raised from tissue cul-
ture as it is expected to have diploid number of chromosomes. The young
1 Cultivation, Conservation and Medicinal Significance of Macrofungi 5
mushroom fruit bodies are first surface sterilized using surface disinfectants such as
mercuric chloride or sodium hypochlorite. The fruit body is then longitudinally cut
into two pieces with the help of sterilized scalpel, and a small piece from the collar
region is carefully transferred onto a suitable culture media such as malt extract agar
(MEA) and potato dextrose agar (PDA) prepared in Petri plates and incubated in a
BOD incubator at 22–28 °C for 5–7 days. The growing mycelium from the margins
of the tissue is carefully transferred onto test tubes that have sterilized culture media
and further incubated at suitable temperatures in an incubator for about 14–20 days
to raise mycelial cultures. Alternatively, pure mushroom cultures can also be
obtained from spores. Most of the mushroom fruit bodies bear millions of spores
which remained covered with pileus. The spore prints can be obtained by placing
mature fruit body onto a sterilized paper or sterilized Petri plate.
Diluted spores are transferred to suitable presterilized culture medium in test
tubes or Petri plates and inoculated at favourable temperatures for 10–14 days to
obtain pure culture. A number of culture media such as malt extract agar, potato
dextrose agar, oatmeal agar, compost extract agar, wheat extract agar, etc. are often
used for preparation of mushroom pure cultures.
Millions of minute spores are produced in mushroom fruit bodies which cannot
be used directly as seed for mushroom production under controlled conditions.
Moreover different mushrooms have different sexuality patterns, and every spore
is not essentially fertile. Therefore, mushroom spawn is normally raised as tissue
culture on suitable cereal grains. The spawn is defined as mushroom mycelia
multiplied on a suitable culture medium. The spawn consists of mushroom myce-
lia and a suitable nutritive medium, which provides food to mushroom during
active growing phase. Culpeper described spawn as mycelium of mushroom.
Until the late nineteenth century, spawn used to be gathered from wild. Before the
advent of modern spawn production technology using cereal grain spawn, virgin
spawn from pastures and meadows; flake spawn by breaking of beds; mill-track
spawn using cattle and horse dung, manure and soil; etc. were used to grow
mushrooms.
The Pennsylvania State University did pioneering research in standardizing
spawn-making process using cereal grains and hold two patents. Subsequently
licences under the patent technology were accessible to other mushroom spawn
laboratories. The process for the production of grain spawn and the fundamentals
have not changed since then; you still need a starter culture, cereal grain. The grains
are sterilized and cooled, and the product is grown out. It is no secret that anyone
can make spawn, just as anyone can grow mushrooms. The traditional spawn labo-
ratories throughout the world use cereal grains as base material for spawn prepara-
tion. The spawn preparation steps are as follows.
6 S. K. Singh and R. Pathak
Various types of cereal grains such as wheat, bajra, rye, jowar and agricultural and
wooden waste are used for mushroom spawn production. Good substrate should not
contain inhibitory compounds; large surface area of substrate should be available
for fungal colonization and should provide essential nutrients required by mush-
room mycelium to grow. Edible quality grains without any pesticide treatment or
infestation (fungal or insect) should be used to avoid contamination and retarded
growth.
For spawn production of oyster, white button and paddy straw mushrooms, cereal
grains are the most suitable substrate, whereas for black ear and shiitake mush-
rooms prefer sawdust-based substrates being wood-rotting fungi. To prepare grain
spawn, the grains are washed in clean drinking water thrice to remove admixtures of
straw, soil and seed of other grasses. These cleaned and washed cereal grains are
then boiled for approximately 20 min. Excess water from the boiled grains is then
removed by drying the grains in shed for few hours. These moist grains are mixed
with 2% calcium sulphate and 0.5% calcium carbonate. The application of these
two chemicals not only provides nutrition to the growing mycelium but also over-
comes the problem of stickiness, lump formation and maintains favourable pH of
the grains near neutral to support the growth of the mushroom mycelium.
About 350 g boiled and treated grains are filled in half kg bottles up to 2/3 volume
and plugged with nonabsorbent cotton. These bottles are then autoclaved at 22 lb
p.s.i. pressure at 126 °C for 1.5–2 h. After cooling a piece of growing mycelium is
aseptically inserted onto grains in bottles, and inoculated bottles are incubated at
22–30°C as per the most favourable temperature for the mycelia growth of the
mushroom to be cultivated for 2–3 weeks. The commercial spawn is prepared from
mother spawn after complete colonization of mycelium of mother spawn on cereal
grains. The most suitable temperatures of mother spawn incubation for Agaricus
and Pleurotus spp. are between 22 and 25 °C and for Calocybe and Volvariella spp.
is around 30 °C.
The commercial spawn is prepared in the same way as described under mother
spawn, but instead of glass bottles, double-sealed polypropylene bags are used for
multiplication of mushroom mycelium. Ideally, 500 g of boiled wheat grains treated
with calcium carbonate and calcium sulphate are filled in each polypropylene bag
and packed by inserting cotton in the polypropylene ring and sterilized in an auto-
clave at 126 °C for a period of approx. 2 h. The bags are left overnight and then
1 Cultivation, Conservation and Medicinal Significance of Macrofungi 7
inoculated with about 50–60 grains from prepared mother spawn and are mixed
thoroughly and incubated in a BOD incubator at 22–30 °C as per the requirement of
the mushroom strain. During incubation, the mushroom mycelium starts spreading
in all directions and covers the entire bag during 2–3 weeks. Ready commercial
spawn is stored at 4 °C until used.
straw in about 18 h. The excess water is then drained off and the straw spread on the
floor and then filled in polythene bags of about 4–5 kg capacity. For larger unit,
instead of drums, bigger cemented blocks with more water holding capacity can be
engaged for chemical treatment of straw. The quantity of the fungicide for chemical
sterilization of substrate is based on per hundred litres of water. Alternatively the
wheat straw can be sterilized by soaking the straw in hot water 65–70 °C for 1–2 h.
The rate of spawn mixed is 2–3% vis-à-vis wet weight of the substrate by thor-
ough mixing or by layer spawning method. Spawned substrate is filled in polythene
bags and tightly closed. About 25–30 holes should be made on all sides of each bag
with 2 side cuts at the bottom to drain excess water. The spawned bags are kept in
incubation room at 25 °C for mycelial growth in a closed room. During this period
fresh air and diffused light are not required, and higher CO2 environment is helpful
in early spawn run. Depending on the substrate and the Pleurotus species used, the
mycelial run is often completed in 18–22 days. Once the mycelial run is complete,
a thick mycelial mat is formed indicating that the macrofungi are ready to fruit. The
polythene coverings of the bags are completely removed, and the blocks are arranged
on shelves and watered at least twice daily. During this period diffused light of low
intensity is required. This can be achieved from sunlight entering through window
or 1–2 bulbs for 3–4 h daily. High relative humidity during cropping period should
be maintained (between 70 and 80%). The fruiting initials start within 3–5 days, and
the fruiting bodies mature within a week. The mature fruit bodies should be regu-
larly harvested and used fresh as vegetable or dried to be used later.
A. bisporus requires well composted materials for its growth. Composting is a com-
plex microbial process which involves chain of microbial succession during com-
posting. The process begins with piling up of pre-wetted ingredients into heap to
start with mesophilic flora to develop and degrade the straw. As the temperature of
the pile rises, mesophilic microbes are replaced by thermophilous bacteria and later
replaced by actinomycetes and help in the decomposition of the substrates. Among
different thermophilic fungi, Scytalidium thermophilum is the most important in
terms of compost production, selectivity and nutrition of A. bisporus (Vijay et al.
1997). Composting is essentially anaerobic process, and the mixture is often turned
to expose every portion of the substrate for aeration. A variety of base material is
used for composting of A. bisporus, i.e. wheat straw, paddy straw, barley, oat, maize
stalks, soybean stalks, mustard stalks, etc. A little quantity of nitrogen is also added
to initiate fermentation; it is supplemented with nitrogen and carbohydrate sources
such as animal manures, urea, calcium ammonium nitrate, ammonium sulphate and
molasses; concentrated meals such as wheat and rice bran, soybean meal, castor
meal, etc.; and mineral sources, i.e. gypsum superphosphates, etc. A number of
formulations are available for compost preparation for long, short and indoor
composting.
1 Cultivation, Conservation and Medicinal Significance of Macrofungi 9
farmyard manure, spent mushroom compost, etc. can also be used. An ideal casing
material should have a neutral pH with low EC. The casing layer should be kept
moist. It takes about a week for complete case run; at this stage air temperatures are
lowered to 15–17 °C and RH around 80–85%, which induces pin heads within
7–10 days. The crop is regularly watered, and mushrooms are harvested for
6–8 weeks.
The only mushroom partially adopted by the farmers is the milky mushroom
(Calocybe indica) which was first cultivated by Purkayastha and Nayak (1981) and
later refined by Doshi et al. (1989, 1993). Its cultivation on wheat/paddy straw sup-
plemented with various organic additives gives about 70% conservation at 25–35 °C
and is catching the imagination of farmers/consumers of west Bengal, Karnataka,
Tamil Nadu, Maharashtra and Uttarakhand, but it needs sustained extension sup-
port. This is a tropical mushroom and has a good scope as it is pure white, grows
well in high temperature ranging from 25–37 °C and is suitable for tropical regions.
The effect of different substrates and casing materials on the growth and yield of C.
indica has been studied by Ruhul et al. (2010). Its keeping quality is excellent and
doesn’t turn brown. The production technology is very simple. This mushroom is
suitable for chutney and pickle production.
1.3 Conservation
Not much organized efforts have been made to conserve the available mushroom
biodiversity ex situ, which require their extensive collection, isolation, pure cul-
tures, identifications and conservation in well-established gene banks as regional,
national and even international facilities. Such a coordinated effort for mushroom
genetic resource conservation has become necessary. In view of the threats like
over-exploitation of wild mushroom and restriction imposed on germplasm
exchange due to patenting, IPR issues, etc., mushroom scientists need to lay more
emphasis on isolating pure culture of wild mushrooms and deposit them in well-
recognized regional and/or national gene bank for their long-term conservation and
future exploitation.
When a new genus or a species of microbe is discovered and described, it is
generally deposited in established germplasm bank. This ensures availability of the
organism for use in future. In addition, microbial strains of industrial importance are
preserved and patented, and their availability became restricted (Jong and
Birmingham 1991). Presently there is no authentic on-the-spot examination; method
is accessible to certify spawn quality; a method of preserving selected strains tested
1 Cultivation, Conservation and Medicinal Significance of Macrofungi 11
and proved desirable is of primary importance. The preservation is not a simple and
routine process as it seems to be, due to degeneration during the preparation or
maintenance. This gradually leads to the loss of desired traits resulting in reduced
recovery, retarded growth and low production and productivity (Chang and Miles
1989; Stadelmann 1986).
Spores produced through asexual processes, i.e. heterothallic or secondary
homothallic species, have genetic differences (Miles and Chang 2004). Although
spores from primary homothallic species are principally genetically similar, they
still exhibit genetic variations, for example, Volvariella (Chang et al. 1981). To
ensure fruiting from single spore of heterothallic species requires mating tests, and
therefore, usually, diploid vegetative mycelia of elite strain are conserved (Snell
1984). Once the pure mycelial cultures are prepared, they can be preserved by dif-
ferent methods as per the requirements of a culture bank, a spawn laboratory and/or
a research organization. Nevertheless, availability of the required equipments and
funds becomes a major limiting factor for mushroom culture preservation for a rela-
tively a long period.
1.3.1 Subculturing
Mineral oil preservation envisages protection to the actively growing mycelial cul-
tures by preventing dehydration and lowering metabolic activity and growth through
reduced oxygen tension. In this storage method, mineral oil is autoclaved for 20 min
at 22 p.s.i. consecutively for 2 days and allowed to cool at ambient temperatures.
Thereafter mycelia cultures prepared in test tubes are filled in this presterilized min-
eral oil covering the culture and the media which disconnect the contact of growing
mycelium with the external environment. Basically, shorter slants require less oil to
cover them. Alternatively, mycelial discs from growing culture in a Petri plate or
wheat grain from mother or commercial spawn can be plunged into sterilized min-
eral oil and may be stored at 4 °C in a refrigerator (Singh et al. 2001a).
In juxtaposition with preservation of the culture at 4 °C in a refrigerator, it is an
efficient way of conserving mycelial cultures. Retrieval of mycelia is easy by using
grain spawn or disc with the help of sterilized needle and draining off as much oil
as possible and streaking the inoculum onto agar in plates or tubes. Tilting the cul-
ture plate facilitates drainage; the excess of mineral oil can also be soaked with the
help of a sterilized filter paper. Subculture at the time of retrieval shows retarded
growth rate, and a second subculture is expected to re-establish the culture.
Contamination by airborne spore and retarded growth on retrieval are two disad-
vantages of mineral oil storage. The method involves cutting 5-mm-diameter discs
from fully grown cultures and storing them at ambient temperature in test tubes in
liquid paraffin and plugged with nonabsorbent cotton. The cultures remained viable
for 8 years.
Boeswinkel (1976) reported that water storage of 650 plant pathogens belonging to
the Phycomycetes, Ascomycetes, Fungi Imperfecti and Basidiomycetes could remain
viable for 7 years. For water storage, the mycelia cultures are raised in Petri plates
1 Cultivation, Conservation and Medicinal Significance of Macrofungi 13
onto a suitable culture media, and small bits of 5 mm diameter are transferred asep-
tically to precooled and sterilized McCartney bottles containing distilled water; the
lids are screwed down tightly and are stored at ambient temperatures.
Most of the mushroom cultures except a few like Volvariella spp. and Calocybe
indica can be stored by this method. Demineralized water without any nutrition
works better. Revival of culture is by removal of a block and placing the mycelium
on a suitable growth medium. Survival of fungal cultures stored in water is reported
for 2–5 years period satisfactorily at IMI, Kew, Survey, UK (Smith and Kolkowski
1996). Freire et al. (2016) evaluated the viability, contamination and morphological
changes of endophytic fungi maintained under different preservation methods.
1.3.4 Lyophilization
the ampoule upon lighting the torch near ampoule. To check the postfreezing count,
any ampoule may be selected at random and break open to check the viability before
finally storing the ampoules for longer durations. If vacuum is perfect, then viability
of most of the organisms does not decrease significantly upon freeze-drying, and
lyophilized cultures can be stored in a refrigerator for several years satisfactorily.
While retrieving the lyophilized culture, it should be re-hydrated in sterile distilled
water for half an hour so as to absorb moisture before inoculating on a suitable cul-
ture medium.
Tan et al. (1991) demonstrated that hyphal cooling at the rate of –1 °C/min to
temperatures of – 45 °C and then −75 °C produced fully freeze-dried mycelia.
Freeze-drying was performed for 2 h at −40 °C, 20 h at −2 °C and 8 h at 20 °C,
resulting in residual moisture content of 2%. Hyphae of Ascomycetes and
Basidiomycetes have been reported to survive freeze-drying. A new lyophilization
protocol has been developed by lyophilization of mushroom mycelium multiplied
on pearl millet grains instead of culture suspension in glass ampoules to improve
survival rates reaching almost 100% (Singh et al. 2004a). Croan et al. (1999)
reported successful lyophilization of basidiomycetes using trehalose solution for
freeze-drying. Lyophilized spores of dictyostelids could remain viable for 30 years
(Raper 1984).
above −139 °C (Morris 1981), and this can cause injury to living cells during stor-
age. Change of physical state from liquid to solid results in increases of volume of
water by 10% and creates mechanical stress inside the cells (Grout and Morris
1987), whereas at −196 °C no recrystallization or movement of molecule has been
reported, and complete dormancy is induced, during which the organism does not
undergo any phenotypic or genotypic change, provided sufficient care is taken dur-
ing freezing of cultures and thawing to recover from preserved culture. This method
can be applied to both sporulating and mycelial cultures. The recoveries of temper-
ature-sensitive strains have been enhanced by standardization of the technique for
individual strain and that have previously failed (Morris et al. 1988; Singh et al.
2004b).
The temperature of nitrogen gas at its vapour phase is 139–140 °C temperature
and that of liquid phase is −196 °C. The mycelia suspension of mushrooms in cryo-
protectant like glycerol (10–15%) or DMSO that can sustain ultra-low temperature
is prepared and distributed in aliquots of 0.5 ml–1 ml in plastic screw cap cryovials
that can withstand ultra-low temperature. Programmed cooling at 1–10 °C per min-
ute is perfect. Alternatively, culture vials are kept in a mechanical freezer main-
tained at −70 °C for 60 min and plunged into liquid nitrogen. Within 24 h the
viability checks before and after freezing of a culture are performed to ensure long-
term preservation of the cultures. Rapid thawing of cultures at 37 °C improves via-
bility (Singh et al. 2001b).
San Antonio (1979) found that cryogenic storage did not affect culture viability
and mushroom production for 9 years. Elliott and Challen (1979) stored 1012 cul-
tures of Agaricus bisporus and related species and reported 95% recovery rate.
Morphological and physiological characters remained unchanged after cryogenic
storage of eight commercial mushroom cultures of A. brunnescens (A. bisporus)
preserved in liquid nitrogen for 10 years (Jodon et al. 1982). Challen and Elliott
(1986) reported successful preservation of species of Agaricus, Coprinus, Lentinula,
Pleurotus, Schizophyllum, Tremella and Polyporus except Volvariella in 10% aque-
ous glycerol solution. They also reported that 10% aqueous DMSO solution gave
constantly reliable retrieval of V. volvacea. Chen (1987) reported that 122 strains
from 42 Basidiomycetes species including important edible mushrooms survived
better with slow cooling (at 1 °C/min.) than rapid freezing. Slow freezing and rapid
thawing generally gave the highest viability count. Ohmasa et al. (1996) recorded
mycelial growth of Flammulina velutipes cultures stored in liquid nitrogen for
7 years. Singh et al. (2004b) preserved mushroom mycelium multiplied on wheat
grains instead of mycelial disc in liquid nitrogen and tested survival, yield and
genetic stability of 11 edible mushroom stock cultures. They modified cryopreser-
vation protocols, gave experimental demonstration of genetic stability of stock cul-
tures and validated the use of liquid nitrogen cryopreservation for long-term
preservation of mushroom cultures. Nevertheless, cryopreservation with liquid
nitrogen, using DMSO as cryoprotectant, has been reported not the most appropri-
ate one for A. blazei preservation (Colauto et al. 2012).
16 S. K. Singh and R. Pathak
Mushrooms contain 20–35% protein with high digestibility which is higher than
most vegetables due to low calories, low fat and high K:Na ratio; mushrooms are the
dietician’s choice for those with obesity and hypertension and suffering from hyper-
acidity and constipation. With low calorie, no sugars and starch and high protein,
they are set to be ‘delight of the diabetic’. Alkaline ash content coupled with high
fibre present in mushrooms is suited to those with hyperacidity and constipation
(Rai and Arumuganathan 2005).
Besides the medicinal significance, the pharmacological research confirms anti-
fungal, antibacterial, antioxidant and antiviral properties of mushrooms (Wani et al.
2010). Nutritionally important chemical components, namely, proteins, amino
acids, carbohydrates, fat, ash, fibre, antioxidants, vitamins, flavour and taste com-
pounds and contents of wild-grown mushrooms, have been reviewed by Wang et al.
(2014).
Ganoderma lucidum, called Rishi mushroom in Japan and Ling Zia in China, is
the most priced medicinal mushroom in the Chinese and Japanese system of medi-
cine but, of late, found appreciable acceptance throughout the world including the
developed countries. It has been found to grow on trees as a phytopathogenic fungus
during monsoon season and found associated with wide range of hosts including
Prosopis spp., Acacia spp., Azadirachta indica, etc. causing economic losses. Its
fruiting bodies have gained wide popularity due to rare medicinal property through-
out the world particularly in Japan, China and the USA. It is reported to possess a
plethora of health and medicinal benefits including significant anticancer, anti-heart
attack and anti-HIV activities and hypoglycaemic, hypotensive, hepato- and nephro-
protective and antioxidant properties (Kim et al. 1996; Liu et al. 1995; Chang 1995,
1996). It is consumed as a tonic and supplement due to its medicinal properties like
antitumour, immunomodulatory, anti-platelet, aggregation, cardiovascular, respira-
tory, anti-hepatotoxic, anti-HIV, hypoglycaemic and anti-nociceptive effects. There
is a big herbal market in American and European countries for this mushroom due
to its health benefits. It is a tropical mushroom fruit under high temperature (30–
38 °C) and high humidity. Its cultivation technology has been developed by the
Directorate of Mushroom Research, Solan, on sawdust substrate (Rai 2003). The
most important bioactive polysaccharide from G. lucidum β-1-3, β-1-6, D-glucan is
high molecular weight polysaccharide and has exhibited significant antitumour
activity. The medicinal component of G. lucidum such as polysaccharides, triter-
penes, organic germanium, etc. and their function have been described by Cheng
and Buswell (1996).
Lentinula edodes generally referred to as Shiitake is considered as medicinal
food mainly in China, Japan and other Asian countries (Yap and Ng 2001). Its cul-
tivation technology has been standardized by Singh et al. (2008). This mushroom
has got as an excellent potential to boost the economy of farmers due to its medici-
nal applications. It possesses anticancerous, anti-hepatitis, cholesterol-lowering,
antihypertensive, antiviral and libido-increasing properties. A bioactive metabolite
18 S. K. Singh and R. Pathak
from L. edodes called lentinan has been reported to be useful in the treatment of
lung, liver, ovarian and stomach cancers (Hazama et al. 1995). It is known to pos-
sess strong immunomodulatory properties. The methanolic extract of Shiitake
mushroom confers protection against hydrogen peroxide-induced cytotoxicity in
peripheral blood mononuclear cells (Kuppusamy et al. 2009).
Phellorinia species is a popular mushroom in Rajasthan, Delhi, Haryana, Punjab
and Uttar Pradesh (Doshi and Sharma 1997). This mushroom is collected in tonnes
and after one or two good monsoon rains from different zones of Rajasthan even
from sand dunes and sold in the market as fresh as well as in dried form. The rural
people have identified the richness of calcium through experience and confirmed
scientifically later. The species of Phellorinia mixed with pure ghee in semisolid
state is given to person suffering from bone crack. It is also given to pregnant women
with local Ayurvedic preparations. Attempts have been made to domesticate
Phellorinia spp. under ICAR ad hoc project by Dr. Anila Doshi and co-workers.
Bioactive components of Podaxis pistillaris exhibited antibacterial activity
(Al-Fatimi et al. 2006). Further production technology has not been standardized
for Podaxis pistillaris.
A lot of Pleurotus species have been found associated with dead and living tree
bark. The cultivation technology of P. sajor-caju, P. sapidus, P. florida and P. citri-
nopileatus and P. flabellatus has been standardized on wheat straw-based substrate
(Anonymous 2009). Fractionation extract of edible mushroom Volvariella volva-
cea, Agaricus bisporus and Calocybe indica yields nicotinic acid carboxylic acid
and ergosterol and correlated with the beneficial health effects as food (Mallavadhani
et al. 2006). Belova and Denisova (2005) studied the possibilities of use of white-rot
wood-destroying fungi for agricultural and industrial waste utilization as a possible
source of energy. Valencia et al. (2006) studied the biological quality of protein
from three strains of Pleurotus spp. Fruit bodies of all the strains possessed high-
protein content, approximately 27% with very high-protein digestibility up to 98%.
Sarangi et al. (2006) identified three natural proteoglycans from Pleurotus ostreatus
mycelia as immune modulators and anticancer agent. Matsubara et al. (2006) dem-
onstrated the potential of white-rot fungi in the bioremediation of contaminated
land. P. eryngii extracts alleviate the decrease in the trabecular bone mineral density
and had significant role in bone metabolism when tested on rats (Kim et al. 2006).
Yi et al. (2006) carried out studies on dietary fibre in P. tuber-regium. Gambato
et al. (2016) evaluated mushroom productivity and antioxidant properties of edible
macrofungi Pleurotus albidus and Pycnoporus sanguineus and found that the sub-
strate ingredients affected mushroom production and chemical composition. Ren
et al. (2014) tested antioxidant and antibacterial activities of eight edible mush-
rooms. They reported that the aqueous extracts of all the mushrooms demonstrated
radical scavenging activities and P. australis with the highest antioxidant activity.
The novel antibacterial compounds from Agaricus bisporus and Pleurotus sajor-
caju were found the most sensitive against Escherichia coli enterobacter spp. and
Pseudomonas spp. (Tambekar et al. 2006). Puttaraju et al. (2006) studied the anti-
oxidant activity in 23 indigenous edible mushrooms. The two mushrooms
Termitomyces heimii and T. mummiformis displayed the maximum antioxidant
1 Cultivation, Conservation and Medicinal Significance of Macrofungi 19
activity potential due to gallic acid, gentisic acid, protocatechuic acid and tannic
acid and developed an important database for the studies of mushroom for prepara-
tion of mushroom-based nutraceutics. Loganathan et al. (2009) performed a relative
study on the anticancer, antioxidant and antimicrobial property of A. bisporus.
Antioxidant activity of three species of wild mushroom Cantharellus has also been
reported from Northwestern Himalaya (Kumari et al. 2011). Guo et al. (2012) found
the macrofungi species Boletus edulis, Boletus regius, Suillus bovinus, Thelephora
ganbajun and Volvariella volvacea exhibited the highest antioxidant capacities and
total phenolic contents. They identified and quantified gallic, homogentisic, proto-
catechuic and p-hydroxybenzoic acid as bioactive compounds which contribute sig-
nificantly to the antioxidant capacities of these macrofungi.
Cordyceps sinensis is native of Himalayan Mountains in India, Tibet and Nepal
at an altitude of 3000–5000 m. It is entomopathogenic in nature and lives as a para-
site on the larvae of Hepialus armoricanus and grows in soil and canalizes caterpil-
lar and forms a fruiting body at the head of the larva. It has got antioxidant
antitumourous and immune-stimulating properties (Singh et al. 2007). Grifola fron-
dosa popularly known as Maitake has been shown to be effective in regulating blood
pressure, constipation and diabetes. Its extract has shown to kill AIDS virus and
anti-HIV activity (Kim et al. 1996). PSK, a top selling anticancer drug extracted
from Coriolus versicolor, accounts for more than 25% of Japan total sales of anti-
cancer drug (Cheng and Buswell 1996).
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