BIBLIOGRAPHY

ESTACIO, MICHAEL JAVIER. MAY 2011. Coconut Water as a Liquid Medium Base
for Lentinula. Benguet State University, La Trinidad, Benguet.

Adviser: Dr. Bernard S. Tad-awan, Ph.D
ABSTRACT
The study was conduced at the Plant Pathology Department of College of Agriculture at
Benguet State University, La Trinidad, Benguet from April 2011- June 2011. The study aimed to
determine the effect of coconut water as culture medium base for liquid culture of Lentinula sp.
Coconut water added with different amounts of molasses and rice bean were evaluated
on its effect on mycelia weight of Lentinula sp.
Coconut water added with 5 grams powdered rice bean and 20 grams molasses as carbon
source gave the highest mycelial yield compared with plain coconut water, rice bean and
molasses.
Use of Potato Dextrose Broth supplemented with 1% urea, 0.3% KH2PO4 and MgSO4
has the highest mycelia yield compared to the use of organic media.

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INTRODUCTION
Mushrooms have long been regarded as effective medicines for the treatment of various
human diseases. The medicinal properties are due to various cellular components and secondary
metabolites, which can be isolated and identified from the fruiting- body, culture mycelium and
culture broth of mushrooms (Tang, 2007).
Mushrooms also show potential for use in waste management. However, as fungi,
mushrooms have life cycles very different from those of green plants (Stamets and Chilton,
1983).
The substrate material is also important. The substrate material is also very important.
The substrate serves mainly as the vehicle that carries the mycelium of the mushroom. The
growth pattern of the mycelium may be influenced by the substrate, as seen by more rapid
growth (Chang and Miles, 2004).
Coconut water is a healthy and one of the best drinks to hydrate. It helps to remove
toxins from the body and aid digestion, coconut have anti-viral, anti-fungal and anti-microbial
properties. Once exposed to air, the liquid rapidly loses most of its nutritional characteristics and
begins to ferment (Anonymous, 2007). Coconut water contains a variety of nutrients including
vitamins, minerals, anti-oxidants, amino acids, enzymes, growth factors and other nutrients. Over
the past two decades, it’s been used extensively as treatment for cholera, influenza and other
infectious diseases that promote dehydration (Lee, 2008). Coconut water is a natural water filter
that takes almost a month to filter each liter of water. The water travels through many fibers
being purified where it is stored away sterile itself (Safron, undated).
Indoor mushroom production demands a much higher level of knowledge, continuous
monitoring and timely manipulation of environmental conditions. Early production methods
Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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involving the use of solid culture medium such as grain and manure have been reported the
growth of vegetative mycelia. These methods require an undesirable extended period of time for
the mycelium to attain a suitable developmental stage and are labor- intensive requiring manual
transfers of aseptic materials as production is scaled up (Holtz and McCulloch, 1994).
A submerged fermentation process using liquid media reduces the time required to
produce mushroom mycelia. These submerged culture technique, however is a not suitable for
commercial mushroom production. The process is characterized by an increase in broth viscosity
with time which can be due to one or more factors, including increase in cell concentration,
changes in growth morp0hology or the production of extracellular products that are alter the
character of the culture fluid (Tang, 2007).
Mycelium can be cultured in a liquid medium (as opposed to a solid like vermiculite/
brown rice flour). In using liquid medium, it is easier to inoculate substrate and start mycelial
growth much faster than using spore (Biotronics, 2000).
Liquid cultures are used to expand mycelium into a liquid solution to inoculate the
chosen substrate. This is similar in using a multi- spore syringe, except the spores have
germinated into network. Since the spores are already germinated, colonization times are
substantially faster and inoculated substrates have an edge over contamination with speed
(Frequently Asked Questions, 2004). Liquid cultures can be economical, as 1cc from spore
syringe can supply with a volume of liquid inoculants which can be used on many jars or bags
(Frequently Asked Questions, 2004).
Information generated in the study will help future producers of liquid culture utilizing
coconut water as based substrate.
Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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The study aimed to determine the effect of coconut water as culture medium base for
liquid culture of Lentinula sp.
The study was conducted at the Plant Pathology Department of College of Agriculture
at Benguet State University, La Trinidad, Benguet from April 2011- June 2011.






































Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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REVIEW OF LITERATURE


Chemical Constituents of Coconut Water
Numerous medicinal properties of tender coconut water were noted (Veekey, undated).
Coconut water was good for feeding infants suffering from intestinal disturbances, oral
rehydration medium, contains organic compounds possessing growth promoting properties,
presence of saline and albumine make it a good drink in cholera cases and many more.
Pradera (1946) also noted the tender coconut water contains sugar which is steadily
increases from 1-5% to about 5-5.5% in the early months of maturation and then slowly falls
reaching about 2% at the stage of the full maturity of the nut.
The tender coconut water contains most of the minerals such as potassium, sodium,
phosphorus, iron, copper, sulfur and magnesium
Coconut water contains amino acids. These composed of 2.41% alanine, 10.75%
ariginine,3.60% aspartic acid,0.97%x 1.17% cystine, 9.76 x 14.5% glutamic acid, 1.95 x 4.18%
leucine,1.95 x 4.57 lysine, 1.21 x 4.1 % proline,1.23% phenylalanine,0.59 x 0.91 % serene, 2.83
x 3.56 % tyrosine (Pradera, 1946).
Tender coconut water contains both ascorbic acids and vitamin of B group. The
concentration of ascorbic acid ranges for 2.2 to 3.7 mg/ml, which gradually diminishes as the
kernel surrounding the water begins to harden (Pradera, 1946). Vitamin of B group in coconut
water containing 0.64 mg/ml Nicotinic acid, 0.52 mg/ml Panthothenic acid, 0.02 mg/ml
Biotin,0.01 mg/ml Riboflavin, 0.003 mg/ml Thiamine (trace), Pyridoxine (trace).
Coconut water composed of 95.5% water, 0.05% nitrogen. 0.56% Phosphoric acid,
0.25% Potassium, 0.69% Calcium oxide, 0.59% Magnesium oxide, 0.5% Iron, 4.71 of total
solid, 0.80 reducing sugar, 2.08 total sugars and 0.62 ash (Pandatal, 1958).

Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Quality of Coconut Water
Coconut water of drinking quality is clear and colorless, with pH of 5 to 5.4 and a Brix
level ( a measurement of sugar concentration) of 5 to 6.5. Per milliliter, it should have a totlal
microbiological count of less than 5,000, less than 10 of coliform bacteria and zero fecal
coliform. A rancid odor indicates that the small quantity of fats in the liquid has oxidized.
Bacteria and yeast are the main microorganisms that threaten coconut water (Food and
Agriculture Organization, 2007). Young coconut water alone contains candida (known as yeast
infection), a single-cell organisms, candida reproduces asexually and thrives on some of the
body’s by- products; dead tissue and sugars from food.

Mushroom Description
According to Stamets (1998), the genus Lentinula was originally conceived by Earle
in 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, gills edges which are often serrated and distinct preference for woodland environment.
Genus Lentinula have flesh composed of dimitic hyphae or irregular or interwoven cells in the
gill trama.
The cap measures 3-6 cm broad, hemispherical then later expanded, light brown to
deep brown, not viscid, margin inflexed. Lamella is white, adnexed, sinuate to adnate then later
separated to stipe. The stipe is cylindrical, 2-5.5 cm long, 10-18 mm thick at the apex and
fibrous. Spores ellipsoid, 5-7 x 2-3.5µm, smooth, hyaline and inamyloid (Chang and Quimio,
1982).


Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Substrate on Mycelial Production
Wasser et al. (2000), reported that the growth of pure mushroom cultures in submerged
conditions like in a liquid medium, permits the acceleration in growth resulting to biomass yield.
The several optimization of the culture medium composition allows the regulation of mushroom
metabolism in order to obtain high yield of mycelia biomass.
Sornprosent and Sangsuwan (2004), also found out that culture mycelia of Ganoderma
lucidum in Potato Dextrose Agar (PDA) was stationary culture for 52 days while in liquid
medium gave the highest dry weight of mycelia of 0.91g/ 1000ml was produced in 25 days.

Carbon and Nitrogen Ratio
The effect of nitrogen is less specific than that of carbon. The carbon- nitrogen ratio
(C:N ratio) is important in fruiting body formation. The C: N ratio obtained by chemical analysis
of fungal cell is approximately 10:1, but substrate carbon is also used for energy and is respired
as CO2.Thus it is, estimated that an amount is converted to cellular material that is similar to the
amount of CO2; consequently, for growth, a C:N ratio of 20:1 is suitable (Miller,2000).

Medicinal Importance of Mushroom
In oriental folk medicine, Lentinula sp is useful in normalizing high blood pressure
and lowering blood cholesterol level. It is also valuable in stimulating immune system to
increase the body’s ability to ward off cancerous tumors, viral infections and chronic fatigue
syndrome. Lentinus sp is the source of a well known anti tumor polysaccharide, lentinan, from
the fruiting bodies or mycelia (Chihara, 1992).



Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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pH of Culture Medium
Several environmental factors affect the growth of mycelia under submerged culture
such as pH and temperature. In submerged culture, the pH of the medium begins to fall rapidly
after commencement of the culture, irrespective of the initial pH range from 3-6 and becomes
about 3.0 within several days. The optimal pH in liquid culture is 5-6 (Chang and Hayes, 1978).

Nutritional Requirement of Fungi
Essential elements required for the growth of fungi includes macro elements:
potassium, sulfur, magnesium, and calcium; micro elements; iron, copper, manganese, zinc and
molybdenum (Griffin, 1993). Nitrogen promotes enzyme activity and ATP production. Fungi
have natural deficiencies on vitamins (Griffin, 1993).
Carbon source provides structural and energy requirements of the fungal cell
concentration of carbon should probably not exceed 2% (Chang and Miles, 2004). Nitrogen is
essential in the synthesis of amino acids, nucleotides and vitamins (Griffin, 1993). Potassium is
commonly supplied as potassium phosphate. A concentration of potassium at around 10-3 M
satisfies the requirement of fungi. Potassium has a role as a co-factor in some enzyme systems
involving carbohydrate metabolism and important in the maintenance of ionic balance (Griffin,
1993; Chang and Miles, 2004).
Among all metabolic minerals, potassium at 0.004 M concentration is adequate for
most of all the fungi (Belgrami, 1978; Griffin, 1993; Chang and Miles, 2004). Magnesium is
essential to all fungi (Chang and Miles, 2004) and its function as enzymes activation and ATP
metabolism statics at 10-3 M (Griffin, 1993: Chang and Miles, 2004).



Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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MATERIALS AND METHODS


The substrates used in the study was sterilized at 20 psi for 1- 2 hours and laid out
following the Complete Randomized Design (CRD). The treatments are replicated 3 times.

Treatments
T01 = Potato Dextrose Broth + 1% urea + 0.3%KH2PO4 + 0.2%MgSO4
T02 = Coconut water
T1 = Coconut water + rice bean
T2 = Coconut water + molasses
T3 = Coconut water + rice bean + molasses
The rice bean was soaked overnight, cooked and then dried. After drying, the rice
bean was macerated to powder form.

Source of Inoculum
A parent culture of Lentinula sp was subcultured by cutting one centimeter square and
then transferred to petri dishes with PDA. The culture was incubated until the mycelia reaches
1cm from inside the peripheral side of the petri dish.

Introduction of Inoculum
Pure culture of Lentinula sp was sectioned into quadrants with a heat sterilized scalpel
and aseptically transferred into the blender containing the substrate sterilized previously.
The blender was placed in power and stirred in 3 seconds burst and pausing for 5
seconds so that the surviving chunks of agar would fall downwards into the blades. Another 3
seconds burst decimates these pieces. This was followed by one more 5 seconds pause followed
Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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by the last 3 seconds. Mycelium blend was pour into a 230 ml jars previously sterilized with
equal volume. The jars were placed in a shaker for 2 hours. After shaking, the jars were placed in
water bath overnight and maintained 27-280C. These were incubated for 7 days to allow the
substrate to be fully colonized with mycelia.

Harvesting Mycelium
Mycelia were filtered using the filter paper allowing the entire liquid medium to
pass through. The mycelial filtrate on the filter paper was dried in the electric stove at low heat
for 15 minutes. The mycelial filtrate was transferred onto the aluminum foil at known weight,
taken from the fresh weight, oven dried at 80oC for 48 hours to get the oven dried weight
(Aspiras and Tad- awan, 2006).

Data Gathered
1. Incubation period (days) - from inoculation to full mycelial colonization of the
substrates.
2. Mycelial fresh weight (g). Mycelia were filtered using filter paper allowing the
entire liquid pass through. The mycelia filtrate on the filter paper was dried at electric stove at
low heat for 15 minutes
3. Mycelial dry weight (g). The mycelia filtrate obtained in data 2 was transferred in
a aluminum foil with known weight and oven dried at 800C for 48 hours and get the oven dried
weight.



Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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RESULTS AND DISCUSSIONS


Effect of Coconut Water with Molasses and Rice Bean
Among of all the media used in the study, control treatment containing the addition of 1
% urea, 0.3% KH2PO4 and 0.2% MgSO4 gave the highest mean weight of 0.12 g (Table 1, Figure
1). However, there was no significant difference between the above treatment with that of
coconut water also gave a good mycelia yield.
Coconut water contains elements such as zinc, selenium, iodine, sulfur, manganese,
boron, molybdenum and others (Lee, 2008). Nutritional facts also cited that it is composed of
95.5% water, 0.05% nitrogen. 0.56% Phosphoric acid, 0.25% Potassium, 0.69% Calcium oxide,
0.59% Magnesium oxide, 0.5% Iron, 4.71 of total solid, 0.80 reducing sugar, 2.08 total sugars
and 0.62 ash (Pandatal,1958). However, addition of 20 grams molasses and 5 grams powdered
rice bean into coconut water increased mycelial weight (Figure 2).
Figure 3a, shows the mycelia of Lentinula sp after oven drying and Figure 3b shows the
mycelia of Lentinula sp taken from the dried mycelia. The mycelium of the dried Lentinula sp is
hyaline.

a
b

a
Figur
a e 1.a) T01- Potato Dextrose Broth + 1% urea + 0.3%

s
KH2PO4 + 0.2 % MgSO4; (b) mycelia of Lentinula sp
s
s
s
s
s
Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011
s
s
s

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Table 1. Mean mycelia fresh and dry weight (g) of Lentinula sp at 7 days after inoculation


TREATMENT MEAN WEIGHT
FRESH DRY
________________________________________________________________________ Potato
Dextrose Broth + 1% urea +
0.3% KH2PO4
+ 0.2 % MgSO4 0.47a 0.12a

Coconut Water 0.28a 0.05b

Coconut Water + 5grams
Rice bean 0.19a 0 .02b

Coconut Water + 20 grams
Molasses 0.26a 0.05b

Coconut Water + 5 grams rice
bean + 20 grams molasses 0.24a 0.11a

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

a
b


Figure 2.a) T3- Coconut water + 20 grams molasses + 5 grams rice bean;
(b) mycelia of Lentinula sp

Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Figure 3a. dried mycelia of Lentinula sp



Figure 3b. Mycelia of Lentinula sp. after drying



Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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pH of the Substrate
The addition of nutrients and fermentation of the substrate significantly lowered the pH.
This is observed after harvesting the mycelia. Decrease of pH is observed on the treatments with
rice bean, which might have fermented on the process and maybe also the coconut water begins
to ferment.
Based on findings of Chang and Hayes, 1978, pH of the submerged culture begins
rapidly fall after commencement of the culture, irrespective of the initial pH. The optimum pH of
the culture media is 5-6.
The lower the decrease of pH the higher the yield of the mycelia. However, there was no
significance difference between the pH of the treatment (Table 2).

Table 2. Mean ph of the substrate before inoculating and after harvesting
TREATMENTS pH

Potato Dextrose Broth + 1% urea + 0.3 %

0.80b
KH2PO4 + 0.2% MgSO4
Coconut Water 0.96b
Coconut water + 5 grams rice bean

1.27a
Coconut water + 20 grams molasses 0.89b
Coconut water + 5 grams rice bean +
20 grams molasses 1.03a
Means in a column followed by a common letter are not significantly different from each other
at 5% DMRT


Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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SUMMARY, CONCLUSION AND RECOMMENDATIONS
Summary
Among all the media used, Potato Dextrose Broth with 1% urea, 0.2% KH2PO4 and
0.3% MgSO4 has the highest mycelial yield.
There is no difference in the dry weight between T01 which is the Potato dextrose Broth
with 1% urea, 0.2% KH2PO4 and 0.3% MgSO4 and T3 which is the Coconut water with 20g
molasses and 5g rice bean.
Favorable environment, such as pH is used for the growth of the mycelia. Achieving
the optimum pH will help the inoculum grow faster. Shaking also help for the production of the
oxygen in an anaerobic condition which is needed by the Lentinula in order to grow.

Conclusion
Coconut water has the potential as a liquid culture medium based for Lentinula sp with
or without the addition of molasses and rice bean. Coconut water contains vitamins and minerals
that can supply the requirements of the mushroom.

Recommendations

Based on the findings, it is recommended:
Further studies in use of coconut water as liquid medium base for the other species of
mushroom and improve the process in the assessment of the purity of the liquid culture.






Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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LITERATURE CITED


ANONYMOUS, 2007. Health Benefits of Coconut Water. Retrieve May 2010 from
www.knowledgebase-script.com

ASPIRAS, J. and B. TAD-AWAN. 2006. Effects of Mineral and Vitamins Sources
on the Mycelial Production of Ganoderma lucidum

BELGRAMI, K.S. 1978, Physiology of Fungi; V. V. Enterprises, India

BIOTRONICS. 2000. Guidelines for Measuring and Reporting EnvironmentalParameters in
Growth
Chambers.
Retrieved
April
2010
from
http://ncr101.montana.edu/Guidelines/guidelines biotronics 2000.htm.

CHANG,S.T. and P. MILES. 2004. Mushroom Cultivation, Nutritional Value, Medicinal Effect,
and Environment Impact, 2nd ed. CRC. Press. USA.Pp.64-67.

CHANG, S.T. and T. QUIMIO, 1982. Tropical Mushrooms: Biological Nature and Cultivation
methods. Hongkong. Polydesign Printing Co; Ltd. Pp. 397-398

CHANG, S.T. and W. HAYES, 1978. The Biology and Cultivation of Edible Mushrooms.
Academic Press New York, San Francisco, London Pp 146-147

CHIHARA, G. 1992. Immunopharmacology of lentinan, a polysaccharide isolated from
Lentinus edodes: Its application as a defense potentiator. Int. J. Orient. Med. 17:57-77

FOOD and AGRICULTURE ORGANIZATION. 2007. Food and Agriculture Organization of
the United Nation, Agriculture and Consumer Protection Department, Retrieved at
www.faop.org/AG/magazine/0701sp1.htm

FREQUENTLY ASKED QUESTIONS, 2004.Psilocybe Mushroom. Retrieved June 2010 from
www.shoomery.com

GRIFFIN, D.H. 1993. Fungal Physiology. 2nd ed. Wiley- Liss Pud., New York. Pp 130-136.

HOLTZ, R.B and J. McCULLOCH. 1994. Process of Production of production of Inoculum.
HPS, Biotechnologies, Inc. Vacaville California

LEE, L. 2008. Nutritional Benefits of Coconut Water. Retrieved April 2011 from
www.litalee.com/shopexd.asp?id.


MILLER, C. 2000. Understanding the Carbon- Nitrogen Ratio. Retrieved June 2010 from
www.acres.usa.com.

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PANDATAL, K. M., 1958. Coconut Water and its uses. Retrieved May 2011 from
www.absoluteastronomy.com/topics/coconut_water.

PRADERA. 1946. Chemical Constituents of Tender Coconut Water. Retrieved April 2010 from
www.geocities.com.

SAFRON,
J.
undated.
Coconut
Benefits.
Retrieved
May
2010
from
www.youngcoconuts.com/benefits.html.

STAMETS, P. 1998. Growing Gourmet and Medicinal Mushroom. Accompanied Guide to The
Mushroom Cultivation. Term Speed Press Berkely. Retrieved Junhe 2010 from
www.kyotan.com.

STAMETS, P. and CHILTON, J. 1983. The Mushroom Cultivator. Agarikon Press, Olympia,
WA. 415p. Retrieved June 2010 from www.attra.org/attra-pub/mushroom.html#top.

STAMETS, P. 1998. Growing Gourmet Mushroom, Term Speed Press. CA. Pp.259-276.

SORNPRASENT, R. and Y. SANGSUWAN. 2004. Effect of Crude from Mycelia of
Ganoderma lucidum Against the Bacterial Growth. Knowledge.biotic.orth/doc.upload.
Retrieved on June 2010

TANG.Y.J. 2007. Submerged Cultivation of Mushrooms, Food Technology, Biotechnology 45
(3) 221-229 (2007). hrcak.srce.hr/file/38111. Retrieved on May 2010

VEEKEY, I. undated. Numerous Medicinal Properties of Tender Coconut Water. Retrieved on
August 2010 from www.veekeyipek.com/Tender_Coconut_Water.htm.

WASSER,S., P.NIEVO, P. SOKOLOV, M. TMORTISMENETSKY, and S. V.
RESHETNIKOY. 2000. The regulation dietary supplements from medicinal mushroom. In:
Science and Cultivation of Edible Fungi. Vol. 1, L. J. D. Griensven (Ed.). Balkerma,
Rotterdam, Netherlands, Rotterdam, Netherlands








Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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APPENDICES
Appendix Table 1. Mycelial fresh weight (g), of Lentinula sp 7 days after inoculation

REPLICATION
TREATMENT I II III TOTAL MEAN_

Potato dextrose Broth
1% urea + 0.3 % KH2PO4
+ 0.2 % MgSO 0.38 0.15 0.87 1.40 0.47

Coconut Water 0.14 0.48 0.23 0.85 0.28

Coconut Water + 5g
Rice bean 0.11 0.08 0.39 0.58 0.19

Coconut water + 20g
Molasses 0.21 0.19 0.37 0.77 0.26

Coco nut Water + rice
Bean + molasses 0.05 0.10 0.57 0.72 0.24



GRAND TOTAL 4.32

GRAND MEAN 0.28



ANALYSIS OF VARIANCE

SOURCE DEGREE SUM MEAN COMPUTED TABULAR F
OF OF OF SQUARE F .05 .01
VARIATION FREEDOM SQUARE

Treatment 4 0.13 0.03 0.60ns 3.11 5.99
Experimental
Error 10 0.58 0.05

Total 14 0.71

ns= Not significant cv= 79.68

Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Appendix Table 2. Mycelial dry weight(g), of Lentinula sp 7 days after inoculation

REPLICATION
TREATMENT I II III TOTAL MEAN

Potato dextrose Broth +
1% urea + 0.3 % KH2PO4
+ 0.2 % MgSO4 0.05 0.04 0.27 0.36 0.12

Coconut Water 0.04 0.07 0.05 0.16 0.05

Coconut Water + 5g
Rice bean 0 .03 0 .04 0.12 0.19 0.02

Coconut water + 20g
Molasses 0.03 0.04 0.09 0.16 0.05

Coco nut Water + rice
Bean + molasses 0.01 0.03 0.31 0.36 0.11

________________________________________________________________________
GRAND TOTAL 1.21

GRAND MEAN 0.07



ANALYSIS OF VARIANCE

SOURCE DEGREE SUM MEAN COMPUTED TABULAR F
OF OF OF SQUARE F .05 .01
VARIATION FREEDOM SQUARE

Treatment 4 0.02 0.005 5.0* 3.11 5.99
Experimental
Error 10 0.01 0.001

Total 14 0.71

ns= Not significant cv= 39.52%





Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Appendix Table 3. Initial pH of the liquid media before inoculation

REPLICATION
TREATMENT I II III TOTAL MEAN

Potato dextrose Broth +
1% urea + 0.3 % KH2PO4
+ 0.2 % MgSO4 7.5 7.2 7.3 22.0 7.33

Coconut Water 6.2 5.9 6.3 18.40 6.13

Coconut Water + 5g
Rice bean 6.3 6.0 6.0 18.30 6.10

Coconut water + 20g
Molasses 5.9 6.0 6.0 17.90 5.96

Coco nut Water + rice
Bean + molasses 5.9 5.8 5.7 17.40 5.80
______________________________________________________________________
GRAND TOTAL 94.0

GRAND MEAN 6.26




ANALYSIS OF VARIANCE

SOURCE DEGREE SUM MEAN COMPUTED TABULAR F
OF OF OF SQUARE F .05 .01
VARIATION FREEDOM SQUARE

Treatment 4 4.47 1.12 14** 3.11 5.99
Experimental
Error 10 0.81 0.08

Total 14 0.71

**= highly significant cv= 4.52%




Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Appendix Table 4.Final pH of the liquid media after harvesting

REPLICATION
TREATMENT I II III TOTAL MEAN


Potato dextrose Broth +
1% urea + 0.3 % KH2PO4
+ 0.2 % MgSO4 6.9 6.8 6.5 20.2 6.73

Coconut Water 5.1 5.5 4.9 15.5 5.17

Coconut Water + 5g
Rice bean 4.8 4.8 4.9 14.5 4.83

Coconut water + 20g
Molasses 4.9 5.2 5.1 15.2 5.07

Coco nut Water + rice
Bean + molasses 4.7 4.8 4.8 14.3 4.77

GRAND TOTAL 79.7

GRAND MEAN 5.31





ANALYSIS OF VARIANCE

SOURCE DEGREE SUM MEAN COMPUTED TABULAR F
OF OF OF SQUARE F .05 .01
VARIATION FREEDOM SQUARES

Treatment 4 7.89 1.97 59.69** 3.11 5.99
Experimental
Error 10 0.33 0.033

Total 14 0.71

**= highly significant cv= 3.42%


Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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CA-UR FORM 9
Benguet State University
COLLEGE OF AGRICULTURE
La Trinidad, Benguet

Date_________
APPLICATION FOR MANUSCRUPT ORAL DEFENSE
Name: MICHAEL J. ESTACIO________________________________
Degree (Major Field): Bachelor of Science in Agriculture (Plant Pathology) _____
Title of Research: COCONUT WATER AS LIQUID CULTURE MEDIUM BASE FOR
LENTINULA .
Date and Time of Defense: JUNE 3, 2011
Place of Defense: AC 107 Building, Benguet State University
Endorsed:
BERNARD S. TAD-AWAN
Adviser, Advisory Committee
(Printed Name and Signature)

APPROVED
ASUNCION L. NAGPALA JOCELYN C. PEREZ
Member, Advisory Committee Member, Advisory Committee
(Printed Name and Signature) and Department Chairperson
(Printed name and Signature)

RESULT OF MANUSCRIPT DEFENSE

Name and Signature Remarks (Passed/Failed)

BERNARD S. TAD-AWAN ____________________
Adviser, Advisory Committee

ASUNCION L. NAGPALA ____________________
Member, Advisory Committee

JOCELYN C. PEREZ ____________________
Member, Advisory Committee
and Department Chairperson
Coconut Water as a Liquid Medium Base for Lentinula / Estacio, Michael Javier. 2011

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Document Outline

• Coconut Water as a Liquid Medium Basefor Lentinula
  • BIBLIOGRAPHY
  • ABSTRACT
  • INTRODUCTION
  • REVIEW OF LITERATURE
  • MATERIALS AND METHODS
  • RESULTS AND DISCUSSIONS
  • SUMMARY, CONCLUSION AND RECOMMENDATIONS
  • LITERATURE CITED

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