American Journal of BioScience
2015; 3(5): 178-182
Published online August 3, 2015 (http://www.sciencepublishinggroup.com/j/ajbio)
ISSN: 2330-0159 (Print); ISSN: 2330-0167 (Online)
Artificial Cultivation of Ganoderma lucidum (Reishi
Medicinal Mushroom) Using Different Sawdusts as
, Miskat Ara Akhter Jahan
, Kamal Kanta Das
, Saurab Kishore Munshi
Department of Microbiology, Stamford University Bangladesh, Dhaka, Bangladesh
Plant Pathology Section, Biological Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhanmondi,
firstname.lastname@example.org (S. Roy), email@example.com (M. A. A. Jahan), firstname.lastname@example.org (K. K. Das),
email@example.com (S. K. Munshi), firstname.lastname@example.org (R. Noor)
To cite this article:
Subarna Roy, Miskat Ara Akhter Jahan, Kamal Kanta Das, Saurab Kishore Munshi, Rashed Noor. Artificial Cultivation of Ganoderma lucidum
(Reishi Medicinal Mushroom) Using Different Sawdusts as Substrates. American Journal of BioScience. Vol. 3, No. 5, 2015, pp. 178-182.
Ganoderma lucidum is a kind of medicinal mushroom possessing anti-tumor, anti-inflammatory, immune-modulatory,
antioxidant and other biological traits which render it to be used as medicinal herbs to combat against variety of diseases. Present
study was designed to implement a suitable method for artificial cultivation of G. lucidum in polypropylene bags with variety of
cheap and readily available substrates. Sawdusts of five woods (Swietenia mahagoni, Dipterocarpus turbinatus, Tectona grandis,
Gmelina arborea and Michelia champaca) were used as substrates and each was supplemented with calcium carbonate (CaCO
either rice or wheat bran for cultivation. T. grandis, G. arborea and M. champaca were not found to provoke the further extension of
mycelial growth and hence the growth was stunted. On the contrary, S. mahagoni and D. turbinatus were noticed to impart
comparatively good yield with biological efficiency. Wheat bran was found to be more efficient as supplement than rice bran.
However, S. mahagoni supplemented with wheat bran provided the best yield of mushroom among the substrates which took 6 days,
33 days and 60 days for the mycelial growth, primordial formation and harvesting, consecutively with the subsequent yields of
235.2 g/kg and biological efficiency of 7.6%.
Ganoderma Lucidum, Reishi, Lingzhi, Medicinal Mushroom, Cultivation, Sawdust
Ganoderma lucidum (Fr.) Karst is a member of fungal
group Basidiomycetes which belongs to Polyporaceae
(Ganodermaceae) of Aphyllophorales. Its fruiting body is
named as “Reishi” in Japanese and “Lingzhi” in Chinese [1, 2].
World-wide Reishi occupies a major source of medicine that
has been used for more than 2000 years [3-5]. Commercial G.
lucidum products are available in various forms, such as
powders, dietary supplements, and tea which are farmed from
different parts of the mushroom, including mycelia, spores,
and fruit body . G. lucidum has been used in Traditional
Chinese Medicine (TCM) as a remedy to treat more than 20
different illnesses which include migraine and headache,
hypertension, arthritis, bronchitis, asthma, anorexia, gastritis
hemorrhoids, hyper-cholesterolaemia, nephritis,
dysmenorrhoea, constipation, lupus erythematosis, hepatitis,
leucopoenia, cardiovascular problems and cancer [3, 6-8].
Besides, reishi or lingzhi also attribute some health benefits
which principally include the control of blood glucose levels,
modulation of the immune system, hepato-protection, and
bacteriostasis . Recent studies on lingzhi have
demonstrated numerous biological activities amongst this type
of mushroom, including anti-tumor, anti-inflammatory,
hepato-protective, anti-microbial, hypotensive, anti-diabetic
and hypolipodemic effects [3, 9, 10].
To meet the gradually increasing demand for G.
lucidum as a natural medicine, commercial cultivation of this
American Journal of BioScience 2015; 3(5): 178-182 179
mushroom has been initiated worldwide, especially in the
tropical Asian countries [4, 11]. As different members of
the Ganoderma genus seek different conditions for growth
and cultivation, and the traditional cultivation technique
takes several months for fruiting body development,
artificial cultivation of G. lucidum has been implemented
using the available substrates such as grain, sawdust, wood
logs and cork residues [3, 11-13]. Several substrates have
been investigated worldwide for the cultivation of G.
lucidum till date [3, 14-18].
Usually in Bangladesh the summer season is considered as
the best time for the cultivation of mushroom [19, 20].
Different environmental factors, oxygen level, and calcium
ion concentration, etc. are also important for the cultivation
[21, 22]. Bangladesh Council of Scientific and Industrial
Research (BCSIR) investigated the efficacy of sawdust
supplemented with rice or wheat bran as substrate, and found
the 9:1 ratio of sawdust and rice bran/wheat bran to be
effective for the cultivation of G. lucidum with elevated
production, even in the large scale. Although extensively
used in several Asian and tropical countries, unfortunately its
application as medicine in Bangladesh is still in scarce.
Commercial-based cultivation of such mushroom in this
country is thus critical to confer its extended medicinal use
as it could be a suitable alternative to synthetic drugs with
less adverse effects. Along these lines, the present study
assessed the best cultivation media for achieving high yield,
biological efficiency, and growth (mycelial, primordial and
fruiting body) rate of G. lucidum using different sawdusts
(Swietenia mahagoni, Dipterocarpus turbinatus, Tectona
grandis, Gmelina arborea, and Mechelia champaca), and the
supplements including rice bran, wheat bran and calcium
) for implementing its large scale
cultivation in Bangladesh.
2. Materials and Methods
2.1. Setting and Sampling
Experiments relating to the present study were carried out
in the laboratory of Plant Pathology Section, Biological
Research Division, Bangladesh Council of Scientific and
Industrial Research (BCSIR), Dhaka, Bangladesh within the
time frame of May 2014 to July 2014. The five types of
sawdusts (Table 1) were collected randomly from different
saw mills of Dhaka city, and the supplements (rice bran, wheat
bran and CaCO
) were collected randomly too from rice mills
and other shops. The sample spawn of G. lucidum was
collected from National Mushroom Development and
Extension Center, Savar, Bangladesh.
Table 1. Plant source of sawdust for substrate base.
Scientific name Family
Mahogani Mahogani Swietenia mahagoni Meliaceae
Gerjan Gerjan Dipterocarpus turbinatus
Segun Teak Tectona grandis Verbenaceae
Gamar Beechwood Gmelina arborea Lamiaceae
Chapa Teak chambul
Michelia champaca Verbenaceae
2.2. Isolation and Obtaining Pure Culture of G. lucidum
To obtain the pure culture of G. lucidum, Potato Dextrose
Agar (PDA) culture or tissue culture method was used. A
small piece of tissue was collected from the fruiting body of G.
lucidum mushroom and placed on the sterilized PDA medium
under aseptic conditions. The inoculated medium was
incubated at 25
C for 7-10 days for sufficient mycelia growth
which was identified on the basis of basidiocarp and
basidiospore morphology. Subsequent sub-culturing was then
performed to obtain the pure culture of G. lucidum mycelium
on PDA which was further added to the substrate for fruiting
body generation [3, 23].
2.3. Substrate Preparation
A total of 10 kg of substrate mixture (65% wet weight and
35% dry weight) was prepared for each category of the 5
sawdusts. Therefore, Moisture level of the mixture was
maintained at 65% (6.5 L) [3, 20, 23]. To achieve 3.5 kg dry
weight of substrate (35% of the mixture), and sawdust and
supplements ratio of 9:1, 3.1 kg (90%) of sawdust was mixed
with 0.3 kg (8%) rice bran or wheat bran and 0.1 kg (2%) of
2.4. Spawn Preparation
A total of ten spawn bags for each of the substrate mixture
were prepared. Each of the ten polypropylene bags (18×25 cm)
were filled with 400 g of the above prepared mixture and
packed tightly. The neck of the bag was prepared by using heat
resistant PVC (Polyvinyl chloride) tube. A hole of about 2/3
deep of the volume was made at the center of polypropylene
bags with sharp end stick for space to put inoculum. The neck
was plugged with cotton and covered with a sterile brown
paper and tied with a rubber band. The packets were sterilized
on an autoclave for one hour at 120
C under 1.5 kg/cm
pressure. Following sterilization, the packets were transferred
into a clean aseptic chamber and were allowed to cool for 24
hours [3, 19, 20, 23].
A piece of pure culture medium containing mycelium (6-7
days old) of G. lucidum mushroom was placed aseptically in
the hole of center of the polypropylene bags and again plugged
with cotton wool in a laminar flow chamber. During
incubation, the inoculated packets were kept in almost dark at
C and transferred to the culture room at 25-32
together with 85-95% relative humidity and 250-350 lux light
. Water was sprayed 4-5 times per day and proper aeration
was maintained in culture house to facilitate the development
of the fruiting bodies [3, 23]. Different parameters such as
days required to mycelial growth, primordial initiation, fruit
body maturation, and harvesting were recorded regularly.
Number of fruiting bodies and dry yield of mushroom
(g/packet) were also calculated. The biological efficiency was
determined for each packet using the formula [3, 19, 23, 24]:
Total biological yield (g)
180 Subarna Roy et al.: Artificial Cultivation of Ganoderma lucidum (Reishi Medicinal Mushroom) Using
Different Sawdusts as Substrates
Biological efficiency (%) = -------------------------- × 100
Total dry substrate used (g)
3. Results and Discussion
Due to the increased rate of resistance and several adverse
effects for toxicity of the synthetic drugs, natural products like
herbs have attracted lots of interest to be a suitable drug
alternatives or sources for new drug discoveries in recent
years [8, 24-37]. G. lucidum has already proved its
competency as medicinally active mushrooms with a wide
range of therapeutic and biological effects [3-5, 7, 9,
meet the huge demand of such mushroom for medicinal
purposes, artificial cultivation using different substrates have
been employed so far round the globe [3, 4, 11-13, 20, 23].
However, in context to economical ease of cultivation, no
significant effort has been initiated to develop a suitable and
cheap cultivation method in Bangladesh till date. Present
study thus endeavored to establish an effective cultivation
technique using sawdusts along with rice bran or wheat bran.
The primordial formation period for S. mahagoni with
rice bran and wheat bran was in 35 days and 33 days,
respectively, which were 40 days and 36 days, respectively
in case of D. turbinatus. The average first harvest days of
fruit bodies of G. lucidum for S. mahagoni with both
supplements (rice bran & wheat bran) were 71 days and 60
days, respectively, which were 90 days and 66 days in case
of D. tarbinatus. In the study conducted by Karma and
Bhatta (2013), it was noted to be up to 92 days . After
obtaining the first harvest of G. lucidum, the second third
and fourth harvest can be obtained from the same sample
within each 18 to 20 days. G. arborea, T. grandis and M.
champaca could not lead to the harvesting of G. lucidum
because of the retarded mycelial growth. Thus, those
sawdust samples could not be suggested to consider as
substrate for the cultivation of mushroom of interest.
Table 2. Comparison of growth (mycelial and primordial) using different sawdust samples supplemented with rice and wheat brans.
3 days 6 days 9 days 12 days 15 days 18 days 19 days
Rice bran 3.5 cm 10.5 cm 14 cm Primordial initiation
Wheat bran 12 cm 14 cm Primordial initiation
- - -
Rice bran 1.2 cm 2.4 cm 4.9 cm 8.7 cm 11.2 cm 13.5 cm Primordial
Wheat bran 2.4 cm 5.8 cm 7.2 cm 10.5 cm 12.5 cm 13 cm Primordial
Tectona grandis Rice bran - - - - 1.2 cm Stunted
Wheat bran - - - 1.4 cm 2.2 cm Stunted
Gmelina arborea Rice bran - - - - 2.1 cm Stunted
Wheat bran - - - - 3.1 cm Stunted
Michelia champaca Rice bran - - - 1.2 cm Stunted
Wheat bran - - - 1.2 cm Stunted
Average data for each of the substrate has been shown.
3.1. Comparison of Growth with Different Types of Sawdust
An equal colonization period was observed with both
supplements (rice bran and wheat bran) in case of D.
turbiratus, T. grandis and M. chmpaca which were 18 days,
15 days and 12 days, consecutively. In case of G. arborea,
colonization period with rice bran and wheat bran was 13
days and 15 days, respectively, which was 9 days and 6 days
for S. mahagoni. But it was observed that, only S. mahagoni
and D. tarbinatus allowed primordial initiation from their
mycelial growth, while other three sawdusts (T. grandis, M.
chmpaca and G. arborea) could not extend the mycelium
growth to primordial formation and the growth was stunted
Growth attained for S. mahagoni up to 14 cm in rice
bran and wheat bran before primordial initiation after 9 days
and 6 days, respectively.
For D. turbiratus, the final mycelial growth was found to be
13.5 cm with rice bran, and 13 cm with wheat bran before
primordial initiation after 18 days (Table 2). Recently, Karma
and Bhatt (2013) in India noticed primordial initiation after 35
days using sawdust as substrate supplemented with rice and
wheat brans, maize flour, and bagasse .
3.2. Effect of Different Types of Sawdust on Yield and
The feasibly of any of the sawdust samples used in this
study was determined through measuring the biological yield
of G. lucidum and biological efficiency of the substrate.
Cultivation of reishi mushroom could only be achieved using
S. mahagoni and D. turbinatus. The yield of 235.2 g/kg with
biological efficiency of 7.6% and yield of 210.9 g/kg with
biological efficiency of 6.8% were attained for S. mahagoni
and D. turbinatus, respectively supplemented with wheat bran
(Table 3). The yield was 132.9 g/kg with biological efficiency
of 4.3%, and 110.4 g/kg with biological efficiency of 3.6% for
S. mahagoni and D. turbinatus, respectively when rice bran
was used as substrate (Table 3).
Karma and Bhatt (2013) found the yield of 570 g/100 kg of the
sawdust used . Azizi et al. (2012) reported the yield of
102.58 g/kg with biological
efficiency of 12.89% using
hornbeam saw dust supplemented with 5% malt extract and 10%
wheat bran . Erkel (2009) also found the highest yield
(63.66 g/kg) and biological efficiency (18.63%) using oak
he used wheat bran as supplement . The
American Journal of BioScience 2015; 3(5): 178-182 181
finding of present study in agreement with previous studies [3,
5, 8, 18, 38], and further suggested that wheat bran would be
suitable supplement for mycelial growth when sawdust being
used as substrate.
Table 3. Effect of different type of supplement and control on yield and
biological efficiency of Ganodarma lucidum.
Sawdust Supplement Yield (g/kg) Biological
Rice bran 132.9 4.3
Wheat bran 235.2 7.6
Rice bran 110.4 3.6
Wheat bran 210.9 6.8
Tectona grandis Rice bran 0 0
Wheat bran 0 0
Gmelina arborea Rice bran 0 0
Wheat bran 0 0
Rice bran 0 0
Wheat bran 0 0
Average data for each of the substrates has been shown
The effect of various kinds of sawdust and supplements on
the yield of G. lucidum was investigated in this study. As
described above yield of G. lucidum varied widely depending
on the kind of sawdust and supplements. Therefore it is
important to use the proper substrate for the commercial
production of G. lucidum. S. mahagoni sawdust with wheat
bran showed highest biological efficiency with better yield
among all treatments. However, marginal difference between
rice bran & wheat bran supplement was observed. As rice bran
and wheat bran are the industrial by-products and are
economically cheaper than other supplements like gram flour,
corn flour etc., these could have better applicability in low
income countries. The present study recommended wheat bran
to be used preferably with S. mahagoni for the commercial
production of G. lucidum. However, rice bran could be used as
an alternative supplement.
Authors thank Plant Pathology Section, Biological
Research Division, Bangladesh Council of Scientific and
Industrial Research (BCSIR), Dhaka, Bangladesh for
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