BIBLIOGRAPHY ANGIE C. JUAN. APRIL 2006. ...

BIBLIOGRAPHY

ANGIE C. JUAN. APRIL 2006. The Effects of Potato Dextrose Agar
Composition and Light Supply on the Growth of Shiitake (Lentinula edodes) Pure
Culture. Benguet State University, La Trinidad, Benguet.

Adviser: Janet S. Luis, PhD.

ABSTRACT

The potato dextrose agar (PDA) composition and light supply were evaluated to
determine their effects on the growth and quality of shiitake pure culture. The study was
conducted from November 2005 to February 2006 at the Plant Pathology Service
Laboratory, Department of Plant Pathology, College of Agriculture, Benguet State
University, La Trinidad, Benguet.

Based from the results of the experiment, the 3 days after inoculation (DAI)
growth of the mycelia of shiitake on PDA gave the fastest mean mycelial growth (3.24
mm) and gave better growth when treated with continuous light at 3.00 mm which did not
differed significantly from alternating light with darkness at 2.67 mm. The PDA
concentrate costs P200 to produce a liter which is more expensive than PDA formulated
from potato, dextrose and agar at P150.

At 14 DAI, the highest mean mycelial growth was observed from the PDA
concentrate medium with 43.42 mm. Lighting did not affect mycelial growth.

In terms of the average mean daily mycelial growth, PDA concentrate gave the
fastest mycelial growth with mean of 3.02 mm. Lighting had no effect on mycelial
growth.

In terms of culture quality, concentrate PDA and PDA formulated from potato,
dextrose and agar had abundant, filamentous, white and thick mycelia. The PDA
formulated from potato, white sugar and gulaman bar had scanty, filamentous, creamy
and thin mycelia.

ii

TABLE OF CONTENTS

Page

Bibliography. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i

Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i

Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii

INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

REVIEW OF LITERATURE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Classification of Shiitake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Description of Shiitake. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Shiitake Cultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4
MATERIALS AND METHODS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
6
Preparation of Culture Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Preparation of Pure Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Statistical Design and Treatments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Data Gathered . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
RESULTS AND DISCUSSION. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Mycelial Growth at 3 DAI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Mycelial Growth at 14 DAI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Average Daily Mycelial Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

Pure Culture Quality . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

SUMMARY, CONCLUSION AND RECOMMENDATION . . . . . . . . . . . . . . 19

Summary . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
iii

Recommendation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

APPENDICES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

iv

INTRODUCTION

Shiitake
(Lentinula edodes) is a popular mushroom in the world. The shiitake,
meaning “mushroom of the shii or oak tree” in Japan, is highly prized in the orient for its
flavor and reported medicinal value. It is a major agricultural commodity in Japan, where
about half the world’s supply of shiitake is produced (Anonymous, 1997).
The shiitake is a traditional delicacy in Japan, Korea and China. For at least a
thousand years, shiitake have been grown on logs outdoors in temperate mountainous
region of Asia. To this day, shiitake is the most popular of the entire gourmet mushroom.
Only in the past several decades have techniques involved for its rapid cycle cultivation
indoors on supplemented, heat-treated sawdust-based substrates (Stamets, 1993).
Most shiitake are produced on natural logs but many farmers in the United
State of America, Taiwan, China, Canada and Singapore are now producing shiitake on
synthetic substrate composed mainly of oak sawdust in fully environment-controlled
buildings providing conditions most favorable to the mushroom at their various stages of
growth (Quimio, 1980).
This study was conducted to gather and provide some valuable information
concerning the effects of potato dextrose agar (PDA) composition as culture medium and
light supply on the growth of shiitake pure culture.
For the laboratory technician, this study helped determine the best factors
affecting the preparation of shiitake pure culture. This study can also serve as basis for
individuals going into shiitake mushroom production.
The study was conducted at the Plant Pathology Service Laboratory, Department
of Plant Pathology, College of Agriculture, Benguet State University, La Trinidad,
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

2

Benguet from November 2005 to February 2006.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

REVIEW OF LITERATURE

Classification of Shiitake
The mushroom has been variously placed in the genera Collybia, Armilloria,
Lepiota, Pleurotus, and Lentinus. At this moment, it is now known as Lentinula edodes.
According to Stamets (1998), the genus Lentinula was originally conceived by
Earle in the 1990’s and resurrected by Pegler in the 1970’s to better define members
formerly placed in Lentinus. Both genera are characterized by white spores, centrally to
eccentrically attached stems, gill edges which are often serrated, and distinct preference
for woodland environment. The genus Lentinula is monomitic, i.e. lacking dimitic
hyphae in the flesh and descending in their arrangement within the gill trama. Members
in the genus Lentinus have flesh composed of dimitic hyphae and highly irregular or
interwoven cells in the gill trama.

Description of Shiitake
Stamets (1998) stated that the shiitake mushroom cap measures 5-25 cm broad,
hemisphere, expanding to convex and eventually plane at maturity. The cap is dark
brown to nearly black at first, becoming lighter brown in age, or upon drying. The cap
margin is even to irregular, in rolled at first, then incurved, flattening with maturity and
often undulating with age. The gills are white, even at first, becoming serrated or
irregular with age, white, bruising brown when damaged. The stem is fibrous, centrally to
eccentrically attached, fibrous, and tough in texture. The flesh bruises brown.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

4

Bassett (Undated) reported that the shiitake and other edible mushroom are
cultivated on a large scale and complete in commercial importance. Its convex cap varies
from white to dark brown and its stem has an upturned ring.

Shiitake Cultivation
Cultivation of this mushroom is a centerpiece of Asian Culture, having
employed thousands of people for centuries. The first written record of shiitake
cultivation can be traced to Wu Sang Kwuang who was born in China during the Sung
Dynasty (960-1127 AD). In 1904, the Japanese researcher Dr. Shozaburo Mimura
published the first studies of inoculating logs with cultural mycelium. Once inoculated,
logs produce six months to a year later (Stamets, 1998).
Bratkovitch and Baugman (Undated) noted that the log culture, although
traditional in Asia, has yet become highly profitable In North America –despite the hopes
of many woodlot owners. However, log culture does generate modest supplementary
income and fits well within the emerging concept of mycopermaculture .In contrast,
Indoor cultivation on sterilized sawdust-based substrates is providing to be highly
profitable for those who perfect the technique. Most successful American growers
employ methods originating in Asia for the cultivation of this mushroom on sterilized
substrates by doubling or tripling the mass of each fruiting block and by “through-
spawning”. The Japanese, Taiwanese, and Thai production systems typically utilized
cylindrically shaped bags filled with one kilogram of supplemented sawdust, which
are top, inoculated. This method gives a maximum of two flushes whereas the more
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

5

massive blocks (2-3 kilograms a piece) provide 4 to 5 flushes before expiring.
Przybylowics and Donoghue (1988) noted that the natural method of cultivation
could be done on hardwood logs, especially oak, sweet gum, poplar, cottonwood, alder,
ironwood, beech, birch, willow, and many other non-aromatic, broad-leaf woods. The
denser hardwood produces for as long as six years. The more rapidly decomposing
hardwoods have approximately 50% lifespan. The fruitwood is notoriously poor for
growing shiitake. Although shiitake naturally occurs on oaks and beeches, the purpose of
this mushroom in hardwood stumps in North America had poor success thus far.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

MATERIALS AND METHODS

Preparation of Culture Medium
Lentinula edodes, being a basidiomycetous fungus was cultured in potato dextrose
agar (PDA). As a factor of the study, PDA was prepared in three ways: as PDA
concentrate, as formulated from potato, dextrose and agar, and as formulated from potato,
white sugar and gulaman bar. As PDA concentrate, 20 g was stirred continuously in 1000
ml of distilled water at boiling temperature until gelling.
In PDA formulated from potato, dextrose and agar, 200 g of peeled potatoes were
boiled in 500 ml distilled water. When the potatoes were cooked, this was filtered and the
filtrate was measured. The distilled water was added in the filtrate to make one liter
capacity. Afterwards, the 20 g agar and the 20 g dextrose were added into the filtrate. The
mixture was heated and continuously stirred while adding the ingredients. When the
medium slightly gelled, it was removed from the flame.
In PDA formulated from potato, white sugar and gulaman bar, the 200 g
peeled potatoes were boiled in 500 ml distilled water. When the potatoes were cooked,
this was filtered and the filtrate was measured and added with distilled water to make one
liter. The dextrose was substituted by white sugar at double weight, i.e. 40 g. Likewise
the agar was substituted by gulaman bar at double weight. The mixture was heated and
was stirred continuously until the gulaman bar and white sugar were fully dissolved.

Preparation of Pure Culture
The healthy mushroom cap was chosen. At the basement of the cap, slices of
tissue were cut at about two to three size using a sterilized blade. After which, there
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

7

were disinfected with one percent chlorox solution in two to three minutes and washed
with 3 series of distilled water. After which, these were blot-dried on sterilized tissue
paper to remove excess water on the mushroom tissue. Aseptically, the cut tissues were
transferred using a sterilized forcep into the Petri plates containing solidified PDA. Plates
were incubated inside the incubation box for about one week until the occurrence of
mycelial growth. After one week of incubation, an agar block containing the mycelia was
transferred aseptically using sterilized wire loop on PDA slants or desired sterilized
containers containing PDA.

Statistical Design and Treatment

The effect of PDA composition as culture medium, and light supply on the growth
of shiitake were determined by using the two-factorial completely randomized design
(CRD) replicated three times. There were nine treatment combinations with four
replicates and three samples per experimental unit. There were a total of 108 culture
plates that were evaluated.
The two factors used were the following: the PDA composition as culture medium
composition, and light supply. The strain that was used in this study was the BSU
selection 505. For factor A (PDA composition as culture medium), these were PDA
concentrate, PDA formulated from potato, dextrose and agar and PDA formulated from
potato, white sugar and gulaman bar. For Factor B (light supply), these were absence of
light, continuous light and alternating 12 hours light and 12 hours darkness. The
treatments used were the following:
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

8

Factor A = P DA Composition

A0 = Potato dextrose agar (PDA) concentrate

A1 = PDA formulated from potato, dextrose and agar

A2 = PDA formulated from potato, white sugar and gulaman bar
Factor B = Light Supply

B0 = No light

B1 = Continuous light

B2 = Alternating 12 hours light with 12 hours darkness

Data Gathered

The data gathered were:

1. Mycelial growth (mm) at three days after inoculation (DAI). This refers to
growth of the mycelium on pure culture measured at three days after
inoculation. This was monitored from the point of inoculation to the margin of
mycelial growth.
2. Mycelial growth (mm) at 14 days DAI. This was measured as the growth of
the mycelium from point of inoculation to the margin of mycelial growth at 14
DAI.
3. Average daily mycelial growth (mm). This refers to the average daily growth
of the mycelium.
4. Pure culture quality. This refers to the amount thickness and mature of
mycelial growth as observed under the microscope. This was
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

9

photodocumented using the micrograph camera.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

RESULTS AND DISCUSSION

Mycelial Growth at 3 DAI

Effect of PDA Composition. Table 1 shows that PDA formulated from potato,
dextrose and agar gave the highest mean mycelial growth at 3 DAI with 3.24 mm. This
was closely followed by PDA concentrate and PDA formulated from potato, white sugar
and gulaman bar with mean mycelial growth of 2.25 mm and 1.92 mm, respectively.

Effect of Light Supply. Under light supply, data show that continuous light and
alternating 12 hours light with 12 hours darkness gave the best mycelial growth with
means of 3.00 mm and 2.67, respectively. Absence of light gave less mean mycelial
growth of 1.74 mm.
Interaction between PDA Composition and Light Supply. No significant
differences were observed between PDA composition and light supply.
Cost of Production. In terms of production cost of PDA/li., formulation from
potato, dextrose, agar (P150) is cheaper than using concentrated PDA (P200). Also,
continuous light supply means P10.00 expense and alternation of light and darkness
means P4.00 expense.

Mycelial Growth at 14 DAI
Effect of PDA Composition. Concentrated PDA shows the highest mean mycelial
growth of 43.42 mm as affected by PDA composition at 14 DAI (Table 2). This was
followed by PDA formulated from potato, dextrose, agar (22.08) and PDA formulated
from potato, white sugar and gulaman bar (12.33) which did not differ significantly from
each other.
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

11

Effect of Light Supply. At 14 DAI, light supply did not affect mycelial growth
which indicates that L. edodes pure culture is not sensitive to light/darkness.
Interaction between PDA Composition and Light Supply. No significant
differences were observed between PDA composition and light supply.
Cost of Production. In terms of the cost of producing PDA, the use of
concentrated PDA means P200.00 expense which is more expensive than formulating it
from potato, dextrose and agar (P150.00) and potato, white sugar and gulaman bar
(P50.00). For light supply, continuous lighting means P72.52 expense and alternate
light/darkness means P36.30 expense which can be gotten rid off by no light (0 expense)
yet equally good mycelial growth.

Average Daily Mycelial Growth

Effect of PDA Composition. Table 3 and Figure 1 shows the average daily
mycelial growth as affected by PDA composition and light supply. The concentrated
PDA gave the fastest mycelial growth of 3.02 mm. It was closely equally followed by
PDA formulated from potato, dextrose and agar and PDA formulated from potato, white
sugar and gulaman bar with means of 1.32 mm and 0.72 mm, respectively. It should be
noted here that the expensive cost of concentrated PDA gives preference for PDA
formulation from potato, dextrose and agar or potato, white sugar and gulaman bar.
Effect of Light Supply. Light supply shows insignificant differences among
treatments. Being so, the expenses of continuous lighting and alternation of light and

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

12

Table 1. Mean mycelial growth at 3 DAI as affected by PDA composition and light
supply

TREATMENT

MEAN MYCELIAL COST OF

GROWTH(mm) PRODUCTION

(PhP)

Factor A = PDA Composition Per Liter PDA

A0 = PDA concentrate

2.25b

200

A1 = PDA formulated from potato,

dextrose and agar 3.24a 150

A2 = PDA formulated from potato,

white sugar and gulaman bar 1.92b 50
Factor B = Light Supply Per Kilowatt-
Hour

B0 = No light
1.74a 0

B1 = Continuous light 3.00a 14.55
B2 = Alternating 12 hours light,

12 hours darkness 2.67a 7.28

CV = 43.57%

Means in a column followed by a common letter are not significantly different from each other at
5% level DMRT

darkness can be saved by using no light.

Interaction between PDA Composition and Light Supply. No significant
differences were observed between PDA composition and light supply.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

13

Table 2. Mean mycelial growth (mm) at 14 DAI as affected by PDA composition and
light supply

TREATMENT

MEAN MYCELIAL COST OF

GROWTH (mm) PRODUCTION

(PhP)

Factor A = PDA Composition Per Liter PDA

A0 = PDA concentrate

43.42a

200

A1 = PDA formulated from potato,

dextrose and agar 22.08b
150

A2 = PDA formulated from potato,

white sugar and gulaman bar 12.33b 50

Factor B = Light Supply Per Kilowatt-
Hour

B0 = No light
23.25a 0

B1 = Continuous light 30.33a 72.62

B2 =Alternating 12 hours light,

12 hours darkness 24.25a
36.31

CV =44.90%
Means in a column followed by a common letter are not significantly different from each other at
5% level DMRT

Cost of Production. In terms of production, the cost of concentrated PDA gave
preference for PDA formulation. In light supply, the expenses of continuous lighting and
alternation of light and darkness can be saved by using no light.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

14

Table 3. Average daily mycelial growth

TREATMENT

MEAN MYCELIAL COST OF PDA

GROWTH PRODUCTION

(mm) (PhP)

Factor A = PDA Composition Per Liter PDA

A0 = PDA concentrate

3.02a

200

A1 = PDA formulated from potato,

dextrose and agar 1.33b
150

A2 = PDA formulated from potato,

white sugar and gulaman bar 0.72b 50

Factor B = Light Supply Per Kilowatt-
Hour

B0 = No light
1.55a 0

B1 = Continuous light 1.98a 3.64

B2 =Alternating 12 hours light,

12 hours darkness 1.53a
1.82

CV = 45.82%
Means in a column followed by a common letter are not significantly different from each other at
5% level DMRT

Pure Culture Quality

Effect of PDA Composition. Table 4 outlines the mycelial growth quality
observed in the treatments. For concentrated PDA and PDA formulated from potato,
dextrose and agar, both had abundant, filamentous white and thick mycelia. On the other
hand, PDA formulated from potato, white sugar and gulaman bar had scanty,
filamentous, creamy and thin mycelia.
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

15

Legend:

PDA Concentrate
No light

PDA formulated from potato,

Dextrose and agar
Continuous light

PDA formulated from potato,
Alternate 12 hours light,

white sugar and gulaman bar
12 hours darkness

3.5

a

3

2.5
a

2
a
a

1.5
b

1

0.5
b

Mean Mycelial Growth (mm)

0

PDA Composition
Light Supply

Treatment

Figure 1. Average daily mycelial growth

Table 4. Pure culture quality of L. edodes grown in PDA culture medium with varying
ingredients

PDA
PDA FORMULATED
PDA FORMULATED
PARAMETER
CONCENTRATE FROM POTATO,
FROM POTATO,
DEXTROSE AND
WHITE SUGAR AND
AGAR
GULAMAN BAR

Amount of Growth
Abundant
Abundant
Scanty

Form
Filamentous
Filamentous
Filamentous

Color of Mycelium
White
White
Creamy

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

Thickness
Thick
Thick
Thin

16

PDA Concentrate
PDA Formulated-
PDA Gulaman-
No light at 3 DAI
Continuous light
Alternate light (12 hours)
at 3 DAI
at 3 DAI
Plate 1. Comparison of L. edodes pure culture on PDA composition and light supply at
3 DAI

PDA Formulated-
PDA Formulated-
PDA Formulated-
no light at 14 DAI
continuous light at
alternate light (12 hours)
14 DAI
at 14 DAI

Plate 2. Comparison of L. edodes pure culture on PDA Formulated and light supply
at 14 DAI

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

17

PDA Concentrate
PDA Concentrate
PDA Concentrate
(No light) at 14 DAI
(Continuous light) at 14 Alternate light at 14 DAI
DAI

Plate 3. Comparison of PDA Concentrate and light supply at 14 DAI

PDA Gulaman - no light PDA Gulaman -
PDA Gulaman - alternate
at 14 DAI
(Continuous light) at 14
light at 14 DAI
DAI

Plate 4. Comparison of PDA Gulaman and light supply at 14 DAI

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

18

Microscopic (40x) hyphal observation
Microscopic (40x) hyphal observation
on PDA Concentrate
on PDA Formulated

Microscopic (40x) hyphal observation on

PDA Gulaman

Plate 5. Microscopic (40x) hyphal observations on PDA composition and light
supply

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

SUMMARY, CONCLUSION AND RECOMMENDATION

Summary

The study was conducted to determine the effects of PDA composition as culture
medium and light supply on the growth of Lentinula edodes pure culture. This was
conducted at the Plant Pathology Service Laboratory, Department of Plant Pathology,
College of Agriculture, Benguet State University, La Trinidad, Benguet from November
2005 to February 2006.

Based on the results of the experiment, the 3 DAI growth of the mycelia on PDA
gave the fastest mean mycelial growth (3.24 mm) and gave better growth when treated
with continuous light at 3.00 mm which did not differ significantly from alternating light
with darkness at 2.67 mm. The PDA concentrate costs P200 to produce a liter which is
more expensive to PDA formulated from potato, dextrose and agar at P150.

At 14 DAI, the highest mean mycelial growth was observed from the PDA
concentrate medium with 43.42 mm. Lighting did not affect mycelial growth.

In terms of the average mean daily mycelial growth, PDA concentrate gave the
fastest mycelial growth with mean of 3.02 mm. Lighting had no effect on mycelial
growth.

In terms of culture quality, concentrate PDA and PDA formulated from potato,
dextrose and agar had abundant, filamentous, white and thick mycelia. The PDA
formulated from potato, white sugar and gulaman bar had scanty, filamentous,
creamy and thin mycelia.
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

20

Conclusion

Based on the results, it could be concluded that L. edodes mycelia grow fastest at
PDA concentrate and does not need light to grow. Considering the cost of PDA
concentrate, the PDA formulated from potato, dextrose and agar is an alternative with
equally excellent mycelial quality effect.

Recommendation

The pure culture of L. edodes is recommended to be isolated in PDA formulated
from potato, dextrose and agar and to be incubated under dark condition.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

LITERATURE CITED

ANONYMOUS, 1997. Shiitake. Retrieved from http://216.55.141.125/shiitake. htm/

BASSETT, B. Undated. Edible and Poisonous Mushroom. Jefferson City,
Missouri.Retrieve from http://www.covection.state. mo.us/mushroom

BRATKOVITCH, S.M. and M. BAUGMAN. Undated. Shiitake Mushroom
Production: Economic Consideration. Facts Sheet Extention. Ohio State University.
Retrieved from http://ohioline.uso.edu.0.0040 htm/

LONGE, J. Undated. Retrieve from http://www.publish. c.s.ro.au/journal/ap

PRZYBYLOWICZ, P. and JOHN DONOGHUE.1998. Shiitake Growers Handbook.
Kendall/hunt Publishing Company.USA.Pp.11, 183-184

QUIMIO, T. H. 1980. Philippine Mushrooms: Survey and Culture of Edible Ones. Final
report on the regional workshop in the cultivation of edible mushrooms in the
tropics. Manila, Philippines. Pp.10-12

STAMETS, P. 1998. Growing Gourmet and Medicinal Mushroom, Ten
Speed Press. CA. Pp. 259-276.

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

APPENDICES

APPENDIX TABLE 1. Mean mycelial growth (mm) at 3 DAI as affected by PDA composition
and light supply (Actual)

TREATMENT REPLICATION
TOTAL
MEAN
I II II IV
Factor A – Culture Medium

Composition

A0 - Potato dextrose
3.00
2.00
1.67
2.33
9.00
2.25
agar (PDA)
concentrate

A1 – PDA formulated
2.33
3.30
3.67
3.67
12.97
3.24
from potato,
dextrose and agar

A2 – PDA formulated

from potato,
2.00
1.67
1.67
2.33
7.67
1.92
white sugar and
gulaman bar

Factor B - Light

B0 – No light
1.67
1.30
2.00
2.00
6.97
1.74

B1 – Continuous light
2.67
2.67
3.00
3.67
12.01
3.00

B2 – Alternating 12

hours light, 12
3.00
3.00
2.00
2.67
10.67
2.67
hours darkness
Grand Total

59.29
Grand Mean
2.47

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

23

APPENDIX TABLE 2. Mean mycelial growth (mm) at 3 DAI as affected by PDA composition
and light supply (Transformed)

TREATMENT REPLICATION
TOTAL
MEAN
I II II IV
Factor A – Culture Medium

Composition

A0 - Potato dextrose
1.87
1.58
1.47
1.68
6.60
1.65
agar (PDA)
concentrate

A1 – PDA formulated
1.68
1.45
2.04
2.04
1.71
1.93
from potato,
dextrose and agar

A2 – PDA formulated

from potato,
1.58
1.47
1.47
1.68
6.20
1.55
white sugar and
gulaman bar

Factor B - Light

B0 – No light
1.47
1.34
1.58
1.58
5.97
1.49

B1 – Continuous light
1.78
1.78
1.87
2.04
7.47
1.87

B2 – Alternating 12

hours light, 12
1.87
1.87
1.58
1.78
7.10
1.77
hours darkness
Grand Total

41.05
Grand Mean
1.71

ANALYSIS OF VARIANCE

Source of
Degree of
Sum of
Mean of
Computed
Tabulated F
Variance
Freedom
Squares
Square
F
.05 .01
Treatment 11 31.4919
2.863 2.47*
2.22 3.09
Block
3
1.2230
0.408
0.35ns

A
2
11.4005
5.700
4.92*
3.40 5.61
B
2
10.2005
5.100
4.41*
3.40 5.61
A*B
4
8.6678
2.167
1.87ns

Error 24
27.7844
1.158

Total 35
59.2764

* - significant
ns
–
not
significant
CV
=
2.62%
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

24

APPENDIX TABLE 3. Mean mycelial growth (mm) at 14 DAI as affected by PDA composition
and light supply (Actual)

TREATMENT REPLICATION
TOTAL
MEAN
I II II IV
Factor A – Culture Medium

Composition

A0 - Potato dextrose
50.64
50.00
31.67
41.33
173.67
43.42
agar (PDA)
concentrate

A1 – PDA formulated
21.33
22.33
25.67
19.00
88.33
22.08
from potato,
dextrose and agar

A2 – PDA formulated
15.00
9.67
9.67
15.00
49.34
12.33
from potato,
white sugar and
gulaman bar

Factor B - Light

B0 – No light
24.00
18.33
25.00
25.67
93.00
23.25

B1 – Continuous light
35.00
34.67
18.67
33.00
121.34
30.33

B2 – Alternating 12
28.00
29.00
23.33
16.67
97.00
24.25
hours light, 12
hours darkness
Grand Total

622.68

Grand Mean
25.94

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

25

APPENDIX TABLE 4. Mean mycelial growth (mm) at 14 DAI as affected by PDA composition
and light supply (Transformed)

TREATMENT REPLICATION
TOTAL
MEAN
I II II IV
Factor A – Culture Medium

Composition

A0 - Potato dextrose
7.15
7.11
5.67
6.47
26.40
6.60
agar (PDA)
concentrate

A1 – PDA formulated
4.67
4.78
5.12
4.92
18.99
4.75
from potato,
dextrose and agar

A2 – PDA formulated
3.94
3.19
3.19
3.94
14.26
3.56
from potato,
white sugar and
gulaman bar

Factor B - Light

B0 – No light
4.95
4.34
5.050
5.12
14.46
4.86

B1 – Continuous light
5.96
5.93
4.38
5.79
22.064
5.52

B2 – Alternating 12
5.34
5.43
4.88
4.14
19.79
4.95
hours light, 12
hours darkness
Grand Total

120.96

Grand Mean
5.04

ANALYSIS OF VARIANCE

Source of
Degree of
Sum of
Mean of
Computed
Tabulated F
Variance
Freedom
Squares
Square
F
.05 .01
Treatment 11 7402.389 672.944
4.96**

Block
3
225.000
75.000
0.55ns
2.22 3.09
A 2
6065.389
3032.694
22.34**

B
2
352.722
176.361
1.30ns
3.40 5.61
A*B
4
759.278
189.819
1.40ns
3.40 5.61
Error
24
3257.500
135.729

Total 35
10659.89

** - highly significant

CV = 9.63%
ns – not significant
The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

26

APPENDIX TABLE 5. Average daily mycelial growth (mm) (Actual)

TREATMENT REPLICATION
TOTAL
MEAN
I II II IV
Factor A – Culture Medium

Composition

A0 - Potato dextrose
3.97
3.50
2.13
2.98
12.08
3.02
agar (PDA)
concentrate

A1 – PDA formulated
1.36
1.36
1.48
1.12
5.32
1.33
from potato,
dextrose and agar

A2 – PDA formulated
0.91
0.52
0.57
0.88
2.88
0.72
from potato,
white sugar and
gulaman bar

Factor B - Light

B0 – No light
1.60
1.21
1.69
1.71
6.21
1.55

B1 – Continuous light
2.36
2.33
1.10
2.12
7.91
1.98

B2 – Alternating 12
1.78
1.83
1.38
1.14
6.13
1.53
hours light, 12
hours darkness
Grand Total

40.53
Grand Mean
1.69

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

27

APPENDIX TABLE 6. Average daily mycelial growth (mm) (Transformed)

TREATMENT REPLICATION
TOTAL
MEAN
I II II IV
Factor A – Culture Medium

Composition

A0 - Potato dextrose
2.11
2.00
1.62
1.87
7.60
1.90
agar (PDA)
concentrate

A1 – PDA formulated
1.36
1.36
1.38
1.27
5.37
1.34
from potato,
dextrose and agar

A2 – PDA formulated
1.19
1.00
1.00
1.17
4.36
1.09
from potato,
white sugar and
gulaman bar

Factor B - Light

B0 – No light
1.45
1.31
1.48
1.49
5.73
1.43

B1 – Continuous light
1.69
1.68
1.26
1.62
6.25
1.56

B2 – Alternating 12
1.51
1.53
1.37
1.28
5.69
1.42
hours light, 12
hours darkness
Grand Total

35.00
Grand Mean
1.46

ANALYSIS OF VARIANCE

Source of
Degree of
Sum of
Mean of
Computed
Tabular F
Variance
Freedom
Squares
Square
F
.05 .01
Treatment
11
41.134
3.739
6.24**

Block
3
1.356
0.452
0.75ns
2.22 3.09
A
2
34.090
17.045
28.45**

B
2
1.515
0.758
1.26ns
3.40 5.61
A*B
4
4.172
1.043
1.74ns
3.40 5.61
Error
24
14.379
0.599

Total 35
55.514

** - highy significant CV = 2.21%
ns
–
not
significant

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake
(Lentinula edodes) Pure Culture / Angie C. Juan. 2006

Document Outline

The Effects of Potato Dextrose Agar Composition and Light Supply on the Growth of Shiitake (Lentinula edodes) Pure Culture.
- BIBLIOGRAPHY
- ABSTRACT
- TABLE OF CONTENTS
- INTRODUCTION
- REVIEW OF LITERATURE
  - Classification of Shiitake
  - Description of Shiitake
  - Shiitake Cultivation
- MATERIALS AND METHODS
  - Preparation of Culture Medium
  - Preparation of Pure Culture
  - Statistical Design and Treatment
  - Data Gathered
- RESULTS AND DISCUSSION
  - Mycelial Growth at 3 DAI
  - Mycelial Growth at 14 DAI
  - Average Daily Mycelial Growth
  - Pure Culture Quality
- SUMMARY, CONCLUSION AND RECOMMENDATION
  - Summary
  - Conclusion
  - Recommendation
- LITERATURE CITED
- APPENDICES