Mycobiology 38(2) : 97-101 (2010)
© The Korean Society of Mycology
DOI:10.4489/MYCO.2010.38.2.097
Effect of Different Substrates and Casing Materials on the Growth and Yield of
Calocybe indica
Ruhul Amin , Abul Khair , Nuhu Alam and Tae Soo Lee *
1
1
1,2
2
Department of Botany, Jahangirnagar University, Savar, Dhaka 1342, Bangladesh
Department of Biology, University of Incheon, Incheon 406-840, Korea
1
2
(Received April 8, 2010. Accepted April 30, 2010)
, a tropical edible mushroom, is popular because it has good nutritive value and it can be cultivated commercially. The current investigation was undertaken to determine a suitable substrate and the appropriate thickness of casing
materials for the cultivation of
. Optimum mycelial growth was observed in coconut coir substrate. Primordia initiation with the different substrates and casing materials was observed between the 13th and 19th day. The maximum length
of stalk was recorded from sugarcane leaf, while diameter of stalk and pileus, and thickness of pileus were found in rice
straw substrate. The highest biological and economic yield, and biological efficiency were also obtained in the rice straw
substrate. Cow dung and loamy soil, farm-yard manure, loamy soil and sand, and spent oyster mushroom substrates were
used as casing materials to evaluate the yield and yield-contributing characteristics of
. The results indicate that
the number of effective fruiting bodies, the biological and economic yield, and the biological efficiency were statistically similar all of the casing materials used. The maximum biological efficiency was found in the cow dung and loamy soil casing
material. The cow dung and loamy soil (3 cm thick) was the best casing material and the rice straw was the best substrate
for the commercial cultivation of
.
Calocybe indica
C. indica
C. indica
C. indica
KEYWORDS : Biological efficiency,
Calocybe indica,
Casing material, Spawn, Substrate, Yield
Calocybe indica, commonly known as milky white mush-
oyster mushrooms.
The production and marketing potential of the milky
white mushroom in Bangladesh is promising. Because of
the high local demand for and export potential of this
mushroom, many private entrepreneurs are interested in
its commercial cultivation. This mushroom requires a temperature of 30~35 C and a relative humidity of 70~80%
for cultivation, which is conducive to the environmental
conditions of Bangladesh [8]. Huge quantities of lignocellulosic residues such as rice straw, wheat straw, mustard
straw, maize straw, waste cotton, water hyacinth, sugarcane bagasse, coconut coir, and kash are generated annually through activities of the agricultural, forest, and foodprocessing industries in Bangladesh. About 30 million
tons of rice straw is produced annually in Bangladesh [9],
which might prove suitable for mushroom cultivation.
Therefore, the present investigation was undertaken to
determine the best substrates and casing materials with
appropriate thickness for the commercial cultivation of C.
indica.
room, grows during the summer in the gangetic plain of
Bangladesh and West Bengal of India [1]. This mushroom is a relatively new introduction from India [2]. Its
robust size, sustainable yield, attractive color, delicacy,
long shelf-life, and lucrative market value have attracted
the attention of both mushroom consumers and prospective growers [3]. C. indica is rich in protein, lipids, fiber,
carbohydrates, and vitamins and contains an abundant
amount of essential amino acids [4].
A substrate is an important substance for growing
mushrooms. Usually, a wide range of diverse cellulosic
substrates are used for cultivating mushrooms. Volvariella
volvacea is grown on banana leaves, bracts of pineapple,
coconut coir, coffee bran, coffee pulp, corn cob, corn stover, orange peel, rice bran, rice straw, sisal bagasse, sugarcane bagasse, and wheat straw [5]. Various agricultural
byproducts are being used as substrates for the cultivation
of oyster mushrooms, including banana leaves, peanut hull,
corn leaves, mango fruits and seeds, sugarcane leaves, and
wheat and rice straw [6]. In Asia, rice straw is widely
used as the substrate for cultivating oyster mushroom [7]
and is also considered the best substrate for yield and high
protein content. Wheat straw is commonly used as a substrate in Europe and sawdust is commonly used as a substrate in Southeast Asian countries for the cultivation of
o
Materials and Methods
Mushroom strain and substrates. C. indica was
obtained from the National Mushroom Development and
Extension Centre (NAMDEC), Savar, Dhaka, Bangladesh.
Coconut coir (Cocos nucifera), kash (Saccharum spontanum), maize straw (Zea mays), rice straw (Oryza sativa),
*Corresponding author <E-mail : tslee@incheon.ac.kr>
97
98
Amin
sugarcane bagasse, sugarcane leaf (Saccharum officinarum),
and waste cotton (Gossypium indicum) were used as substrates in this study.
Preparation of spawn. The substrates were chopped into
3~4 inch lengths. On a dry-weight basis, 0.2% CaCO and
30% wheat bran were added to the chopped substrates
and mixed thoroughly. Water was added to constitute a
moisture level of 65%. Substrate (500 g) was added to
polypropylene bags (7 × 10'' sizes), and the openings of
the bags were plugged with cotton and secured with plastic rings. The bags were autoclaved at 121 C at 15 psi for
1 hr, after which they were inoculated with 2 teaspoons of
a mother culture of C. indica. The spawn packet was kept
in a dark room for incubation at 30~32 C for approximately 30 to 35 days.
et al.
can’s multiple range test. Biological efficiency was measured using the following formula:
Biological efficiency (%)
= total biological yield/total substrate used × 100
3
o
o
Casing materials and experimental conditions. Cow
dung and loamy soil (3 : 1, v/v), farm-yard manure, loamy
soil and sand (3 : 1, v/v), and spent oyster mushroom substrate (saw dust) were used as casing materials, which
were collected from the NAMDEC farm. All casing materials were sterilized at 65 C for 4 hr. After mycelial colonization, the mouth of the spawn packet was covered with
casing materials and maintained with five different (1 to 5
cm) levels of thickness. It was then transferred to the culture house and maintained at a temperature of 30~35 C
and a relative humidity of 70~80%.
o
o
Experimental design. The experiment had a completely
randomized design with four replications. The following
data were collected: mycelial growth in the spawn, the
number of days required for the initiation of primordia,
the number of days required for total harvest, length and
diameter of the stalk, the diameter and thickness of the
pileus, the number of effective fruiting bodies, biological
yield, economic yield, and biological efficiency. The data
were analyzed according to standard methods using the
MSTAT-C program. Means were compared using Dun-
Results and Discussion
Effect of different substrates on yield and yield-contributing characteristics. The optimum mycelial growth
in the spawn packet was observed in the coconut coir substrate (0.81 cm), which was statistically similar to that in
the maize straw and sugarcane leaf substrates. The lowest
level of mycelial growth was found in the waste cotton
substrate (0.37 cm). The shortest time required to complete mycelial growth was observed in the sugarcane bagasse
substrate (27.75 days), followed by the maize straw (29.50
days) and sugarcane leaf (29.75 days) substrates. The
longest time (40.25 days) required to complete mycelial
growth was observed in the waste cotton substrate (Table
1). These findings are similar to those of previous studies
using C. indica [10]. The presence of the right proportion
of α-cellulose, hemi-cellulose, pectin, and lignin was the
probable cause of the higher rate of mycelium in the
coconut coir substrate. A suitable ratio of carbon to nitrogen might have been responsible for the higher mycelial
growth. The capacity of mushrooms to grow on lingo-cellulosic substrates is related to the vigor of their mycelium
[11]. The minimum time for primordia initiation was
observed in waste cotton substrate (13.0 days), which was
statistically similar to that of the coconut coir, maize straw,
and rice straw substrates. The longest time was recorded
in sugarcane bagasse substrate (19.00 days). Patra and Pani
[12] reported that the time required for primordia initiation of C. indica on paddy straw was 13~16 days. Similar findings were also reported by Jiskani et al. [13].
Substrates containing glucose, fructose, and trehalose produced the highest number of primordia, whereas abnormal fruiting bodies were produced in glycerol, xylose,
Effect of different substrates on the mycelial growth, fruiting body formation, and yield of
Mycelial growth
Complete mycelial
Primordia
No. of effective
Substrates
in spawn
growth in spawn
initiation
fruiting body
(cm/day)
(days)
(days)
Table 1.
Coconut coir
Kash
Maize straw
Rice straw
Sugarcane bagasse
Sugarcane leaf
Waste cotton
CV (%)
Calocybe indica
0.81a
0.67b
0.72ab
0.71b
0.69b
0.72ab
0.37c
9.30
32.50b
33.00b
29.50cd
32.00b
27.75d
29.75c
40.25a
03.86
13.50d
17.75b
13.75d
14.00d
19.00a
15.75c
13.00d
05.21
3.50d
3.50d
7.00ab
7.75a
3.50d
4.75c
6.00b
13.75
Biological
yield
(g/packet)
346.0d
315.8e
452.3b
472.5a
259.3f
342.8d
401.3c
001.72
Economic
yield
(g/packet)
323.8d
297.3e
427.3b
448.5a
240.3f
324.0d
377.5c
001.49
In a column the same letters indicate that the values are not significantly different by Duncan’s multiple range test (P > 0.05). CV, coefficient
of variation.
Effect of Different Substrates and Casing Materials of Calocybe indica
Fig. 2.
Fig. 1.
The fruiting body of
different substrates.
Calocybe
indica
produced in
sucrose, and fructose. The optimal fruiting body production occurred on glucose and fructose containing substrates [14]. Fruiting bodies of
were formed in
six different substrates (Fig. 1). The maximum biological
and economic yield was obtained from rice straw substrate
(472.5 and 448.5 g). The lowest biological and economic
yields (259.3 and 240.3 g) were obtained from sugarcane
bagasse (Table 1). Of all the substrates tested, the total
length of harvesting time varied from 65.75 to 91.75 days.
The longest time for harvesting was observed in rice
straw substrate (Table 2). The maximum diameter of stalk
and pileus and thickness of pileus were also obtained from
rice straw. Statistically similar stalk lengths were found in
all of the substrates. The minimum diameter (4.98 cm)
and thickness (1.40 cm) of pileus was observed in sugarcane bagasse (Table 2). Biological efficiency varied significantly between the substrates. The highest (91.75%)
and lowest (51.86%) biological efficiencies were observed
in the rice straw and sugarcane bagasse substrates, respectively (Fig. 2). The findings of the present study are comparable with those of previous studies using
[15,
16]. Rice straw was the best substrate, followed by wheat
straw. Therefore, cellulose-rich organic substrates are suitable for cultivating mushrooms [17, 18].
99
Effect of different substrates on the biological efficiency
of
. The vertical bars represent the
standard error. S-1, coconut coir; S-2, kash; S-3, maize
straw; S-4, rice straw; S-5, sugarcane bagasse; S-6,
sugarcane leaf; S-7, waste cotton.
Calocybe
indica
C. indica
C. indica
Fig. 3.
Effect of different casing materials on the biological
efficiency of
. The vertical bars
represent the standard error. Cm-1, cow dung and soil;
Cm-2, farm-yard manure; Cm-3, soil and sand; Cm-4,
spent mushroom substrate.
Calocybe
Effect of different casing materials and thicknesses on
yield and yield-contributing characteristics. Cow dung
and loamy soil, farm-yard manure, loamy soil and sand,
and spent oyster mushroom substrate were used as casing
materials to evaluate the yield and yield-contributing char-
Effect of different substrates on the yield-contributing characteristics of
Total days of
Length of
Diameter of
Substrates
harvest
stalk (cm)
stalk (cm)
Coconut coir
89.75ab
07.90a
2.40bcd
08.38a
1.71f
Kash
76.25e
08.60a
2.68ab
Maize straw
86.00bc
Rice straw
91.75a
09.03a
2.83a
08.77a
1.73f
Sugarcane bagasse
65.75f
09.26a
2.43bcd
Sugarcane leaf
80.25de
06.52b
2.45bc
Waste cotton
83.25cd
CV (%)
03.87
10.89
9.27
Table 2.
indica
Calocybe indica
Diameter of
pileus (cm)
5.66bc
6.11ab
6.09ab
6.53a
4.98d
5.14cd
6.28ab
7.32
Thickness of
pileus (cm)
1.70de
1.68de
2.08abc
2.10abc
1.40e
1.88bcd
1.68de
8.26
In a column the same letters indicate that the values are not significantly different by Duncan’s multiple range test (P > 0.05). CV, coefficient
of variation.
100
Amin
et al.
Effect of different casing materials on fruiting body formation and the yield of
Primordia
Total days
No. of effective
Casing materials
initiation (days)
of harvest
fruiting body
Cow dung and soil
15.25b
67.92bc
05.33a
04.92b
Farm yard manure
14.17c
68.83b
06.08a
Soil and sand
15.17b
71.75a
Spent mushroom substrate
17.67a
66.00c
05.75a
03.98
10.12
CV (%)
6.17
Table 3.
Calocybe indica
Biological yield
(g/packet)
314.7a
313.2a
307.3a
308.3a
002.12
Economic yield
(g/packet)
311.5a
310.6a
305.3a
305.8a
002.14
In a column the same letters indicate that the values are not significantly different by Duncan’s multiple range test (P > 0.05). CV, coefficient
of variation.
Effect of different casing materials on the yield-contributing characteristics of
Casing materials
Length of stalk (cm) Diameter of stalk (cm) Diameter of pileus (cm) Thickness of pileus (cm)
02.01aa
Cow dung and soil
7.55b
2.73c
7.20b
Farm yard manure
8.44a
2.97b
7.55a
02.01aa
Soil and sand
6.57c
2.33d
6.75c
01.83ba
Spent mushroom substrate
8.85a
3.18a
6.13d
01.91ab
9.12a
7.75a
10.11aa
CV (%)
6.63a
Table 4.
Calocybe indica
In a column the same letters indicate that the values are not significantly different by Duncan’s multiple range test (P > 0.05). CV, coefficient
of variation.
Effect of different thicknesses of casing materials on the fruiting body formation and yield of
Thickness of casing
Primordia
Total days
No. of effective
Biological yield
material (cm)
initiation (days)
of harvest
fruiting body
(g/packet)
01.50c
123.5e
1
17.25c
44.00c
05.75b
330.3d
2
18.00c
53.25b
3
20.25b
62.00a
07.50a
375.5c
07.50a
411.5b
4
21.75ab
60.25ab
07.75a
433.8a
5
23.25a
64.75a
07.38
08.60
15.21
002.66
CV (%)
Table 5.
Calocybe indica
Economic yield
(g/packet)
121.5e
327.0d
373.5c
408.8b
431.5a
002.71
In a column the same letters indicate that the values are not significantly different by Duncan’s multiple range test (P > 0.05). CV, coefficient
of variation.
Effect of different thicknesses of casing materials on the yield-contributing characteristics of
Thickness of casing
Length of stalk
Diameter of stalk
Diameter of pileus
material (cm)
(cm)
(cm)
(cm)
02.68c
2.39c
5.05db
1
2
07.75b
2.70b
5.55cd
09.51a
2.72b
6.08bc
3
4
09.28a
2.83b
6.58bb
5
09.42a
3.05a
7.75ab
CV (%)
12.71b
5.34b
7.29bb
Table 6.
Calocybe indica
Thickness of pileus
(cm)
01.33bb
01.57ab
01.65ab
01.71ab
01.82ab
11.76bb
In a column the same letters indicate that the values are not significantly different by Duncan’s multiple range test (P > 0.05). CV, coefficient
of variation.
acteristics of C. indica. The results indicate that the number of effective fruiting bodies, the biological and economic
yields, and biological efficiency were statistically similar
in all of the casing materials tested. Maximum biological
efficiency was recorded in cow dung and soil (62.94%)
followed by the farm-yard manure (62.64%), spent mushroom substrate (61.66%), and soil and sand (61.46%) casing materials (Fig. 3). Primordia initiation occurred earlier
in farm-yard manure than in the other casing materials
(Table 3). The greatest stalk length and diameter were
observed with the spent mushroom substrate casing material. The maximum diameter and thickness of pileus were
found with the farm-yard manure (Table 4). These findings are comparable with data from a study using Agaricus bisporus, because it grows well in composting casing
materials [19]. The effects of five different thicknesses of
cow dung and loamy soil as casing materials on the yield
and yield-contributing characteristics of C. indica are pre-
Effect of Different Substrates and Casing Materials of Calocybe indica
Fig. 4.
Effect of different thicknesses of casing materials on
the biological efficiency of
. The
vertical bars represent the standard error.
Calocybe
indica
sented in Tables 5 and 6. The results indicate that 3 to 5
cm thicknesses of casing materials had significant positive results on the number of days required for primordia
initiation, the total number of days to harvest, the number
of effective fruiting bodies, biological and economic yields,
stalk length and diameter, and diameter and thickness of
pileus. The highest and lowest biological efficiencies were
5 cm (86.76%) and 1 cm (24.7%), respectively (Fig. 4).
The casing layer is an essential component for the artificial cultivation of
. According to Sassine
[20], the casing layer must be very loose; otherwise, the
primordia cannot penetrate from the bottom to the top of
the casing layer. The 3 cm casing layer thickness seemed
to be the most efficient. The combination of composted
cow dung and loamy soil proved to be a good casing
material, which likely played a role in stimulating the initiation of the fruiting body [21-23]. Thus, the 3 cm thick
composting cow dung and loamy soil was the best casing
material and the rice straw was the best substrate for the
commercial cultivation of
.
C. indica
et al.
C. indica
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