Republic of the Philippines

Calamba City Schools Division

CALAMBA CITY SCIENCE HIGH SCHOOL
Chipeco Avenue, Barangay 3, Calamba City, Laguna, 4027

Mycelial growth of Oyster Mushroom (Pleurotus ostreatus) in Arrowroot

(Maranta arundinacea) dextrose agar

An Investigatory Project

Presented to the Faculty of Calamba City Science High School

In partial fulfilment of the requirements

In Research 10

By

Francesca Marie O. Supeña

Researcher

Rowena L. Acosta

Research Adviser

March 2019
 Introduction

Background of the Study

Oyster mushroom cultivation has long been in the industry of agriculture but it

gained popularity nowadays in the Philippines because of its impact in the society, economy, and

environment (Andam and Dulay, 2017). It is said by Chang and Wasser (2017) that mushrooms

can be used as food, tonics, medicines, cosmeceuticals, and as organic biocontrol agents in plant

protection with insecticidal, fungicidal, bactericidal, herbicidal, nematocidal, and antiphytoviral

activities. It can also reduce agricultural and industrial waste because it uses different kinds of

substrates such as sawdust, manure, coir, vermiculite, coffee grounds and more. Mushrooms can

be of great help in our community as they have great potential for generating a socio-economic

impact in human welfare on local, national, and even global levels if they are known by many.

In a study by Dr. Viyar last 2017, they aim to make Laguna the mushroom capital of the

country by introducing its way of culture. Since mushrooms don’t have seeds, the person needs

to create a spawn first by culturing its myceliain an agar. The most common agar used is potato

dextrose agar and other substitutes for potatoes as long as it has a reasonable amount of starch.

This study aims to replace the potato starch with arrowroot starch to know if it can be

used as an alternative starch source for the agar. According to Bryan (2017), arrowroot powder

has more starch than potato which means that fewer amounts can be utilized. It is also a healthier

option than other starches since it is grain-free which means it is a big help for people who has a

hard time digesting, as said by (Dr. Mercola, 2016). According to Capiña (2017), arrowroot

powder has one of the purest form of natural carbohydrates which is considered gluten free and
superior quality starch. These health benefits may help in utilizing it as a starch in agar since it

will be used as the culture media for oyster mushroom spawn.

Statements of the Problem

1. What is the effect of arrowroot (M. arundinacea) starch to the mycelial growth of oyster

mushroom (P. ostreatus) in terms of colonial diameter?

2. Is there any significant difference between the colony diameters of the oyster mushroom

mycelia in Arrowroot dextrose agar and Potato dextrose agar?

Hypotheses

Null Hypotheses

1. There is no significant difference between the colony diameters of the oyster mushroom

mycelia in Arrowroot dextrose agar and Potato dextrose agar

Alternative Hypotheses

1. There is a significant difference between the colony diameters of the oyster mushroom

mycelia in Arrowroot dextrose agar and Potato dextrose agar

Significance of the Study

This study will be conducted to use Arrowroot (M. arundinacea) starch for the culturing

of the mycelia of oyster mushrooms (P. ostreatus). This aims to find an alternative starch for the

culturing of the oyster mushrooms (P. ostreatus) mycelia instead of potato to know if it has more

benefits than the usual Potato Dextrose Agar (PDA).

 The beneficiaries of this study will be the oyster mushroom farmers since they’ll be able

to use another starch that might have more benefits than potato. It’ll also help arrowroot farmers

since they’ll be able to sell arrowroot starch to people who want to plant oyster mushrooms. It

will also benefit the community since oyster mushrooms are alternatives for many kinds of food

and it is healthier than most commercial food like meat and such.

Scope and Delimitation

The study will focus on the mycelial growth of oyster mushroom in Arrowroot Dextrose

Agar in terms of colonial diameter. It will not include any other variables. The study will only

help locals especially oyster mushroom and arrowroot farmers and nothing more. It will only be

conducted for 6 months.

Conceptual framework

This figure indicates the framework for the whole study. In this framework the

dependent variables are stated

Data Gathering
(Colonial Diameter)
Data Analysis
Input: (t-test for independent
Arrowroot starch, groups)
sugar and agar Dependent variable
powder (Colonial diameter
compared to PDA)

Process
Boiling of
Arrowroot starch, Output
sugar and agar Mycelia in the culture
powder in 1 liter media (PDA as control
water and and Arrowroot Dextrose
sterilization Agar as experimental)
 In this conceptual framework, the dependent variable which is the colonial diameter of

the oyster mushroom mycelia will be computed from the data that will be gathered in the testing

of the Arrowroot Dextrose Agar and Potato Dextrose Agar. For the t-test for independent groups,

the ability of the oyster mushroom to grow in the two different culture media will also be

computed.

Definition of Terms

Dextrose –sugar; provides carbohydrates for the culture media

Agar – thickener for the culture media

Mycelia – the vegetative part of a fungus; ‘seeds’ for mushroom

Colonial diameter – the diameter of the growth of the mycelia (measured with a ruler, average of

two perpendicular lines)

 Review of Related Literature and Studies

Mushroom

According to Zelman (2018), mushrooms are an all-natural multivitamins which

obviously has many vitamins, minerals, and nutrients in it such as B vitamins, copper, folate

magnesium, zinc, selenium, and ergothioneine. It also has protein, fiber, potassium, vitamin D,

calcium, and more even though it is more than 90% water. It is also anti-bacterial, anti-oxidant,

good for the immune system, and may even help prevent diseases such as Parkinson’s disease,

Alzheimer’s disease, high blood pressure, and cancer.

Mushrooms are also low-carb, contains less fat, and it has low calories but rich in

vitamins as said by Shortsleeve (2019). Nearly 70-80% of mushroom species are poisonous but

the rest are healthy for humans such as cremini mushrooms, horse mushrooms, button

mushrooms, Portobello mushrooms, and oyster mushrooms.

Oyster Mushroom

Oyster mushrooms are saphotrophs that feed on decaying matter especially wood. Since

it decays matter, it eats up wood, paper, coffee grounds, and some petroleum products which

spikes up interests to use it for decreasing all kinds of pollution. Its mycelia also have the ability

to kill and eat small roundworms and bacteria making it one of the few carnivorous mushrooms.

It is also a powerful absorbent of mercury and mycologists have already used them in oil spills.

(Mushroom Appreciation)

Oyster mushrooms and shiitake mushrooms have the most fiber (at 2g per serving),

Lemond says. It is also considered as a medicinal mushroom since it has many health benefits

according to Busch (2018).

Health Benefits of Oyster Mushroom

Since oyster mushrooms are low in calories, fat-free, cholesterol-free, gluten-free, very

low in sodium and it has high amounts of protein, fiber, iron, zinc, potassium, phosphorus,

selenium, calcium, folic acid, niacin, iron, Vitamin B, Vitamin C, and Vitamin D, it is used for

lowering cholesterol, antioxidant and anticancer. (Sayner, 2018).

According to Stamets, Oyster mushrooms are considered the most famous edible

mushroom because it has many health benefits and it is easy to cultivate. It also helps in blood

circulation through its iron and protein content, boosts metabolism through niacin, calcium

absorption in the body through Vitamin D, and lowers systematic inflammation through its

antioxidant properties given by selenium and ergothioneine. It is also considered as the

mushroom that’s easiest to cultivate because it grows in a wide range of substrates. (GroCycle)

Oyster Mushroom cultivation

The mycelia will be made into oyster mushroom spawn and mixed with a substrate.

These substrates can be coffee grounds, straw, sawdust, and more. It’s placed in bags with a hole

to let air in it. It is then placed in a dark and warm room for the incubation period for 10-14 days

for the mycelia to colonize and grow.

Once it is colonized and inoculated, the oyster mushrooms will start to grow in the

fruiting period. The fruiting bags must be exposed to fresh oxygen, high humidity, low level

light, and cooler temperatures.

Oyster Mushroom Mycelia culturing

To culture oyster mushroom mycelium which is used to spawn it, the person needs to

prepare media plates. The most common one is the Potato Dextrose Agar which is prepared

through boiling potatoes to get its starch. After this, it will be mixed with agar powder at a 1:1
volume ratio in water. After this, sterilization will occur through an autoclave at 121°C, 15 psi

which will be done for 30 minutes. It’ll then be put in petri dishes or any glass and covered with

a parafilm or tape to avoid contamination and then it’ll be ready for the tissue of the oyster

mushroom to produce mycelium. (Hsu et. al, 2018?)

Starch Dextrose Agar

Starch Dextrose Agar is used in culturing of microorganisms such as in fungi and

bacteria. It is composed of starch from plants, sugar or dextrose, and agar powder. According to

Rijal last 2015, the starch serves as the nutrient base, the dextrose serves as the growth stimulant,

and the agar serves as the solidifying agent. The most common example of this is Potato

Dextrose Agar (PDA)

PDA is a medium for the identification, cultivation and enumeration of yeast and molds

in foods and dairy products. It may also be used for the cultivation of yeasts and molds from

clinical specimens. Since it stimulates sporulation and pigmentation, it also aids for cultivating

and differentiating pathogenic and non-pathogenic fungi.

Other starches can also be used in preparing culture media such as cornstarch, starches of

legumes, and literally any starchy vegetables but they have different effects since they have

different starch contents.

Arrowroot Starch

Arrowroot powder is a starchy substance that’s extracted from the root of a tropical plant

known as Maranta arundinacea. When the arrowroot is harvested, it looks similar to other

underground tubers such as cassava, yucca or kudzu, which are oblong in shape. (Bryan 2017)
 According to Capiña (2017), arrowroot powder has one of the purest form of natural

carbohydrates which is considered gluten free and superior quality starch.

Arrowroot Starch vs. Potato Starch

Potato starch, tapioca (made from manioc root), and arrowroot are larger-grained starches

that gelatinize at relatively lower temperatures (Christensen, 2009). According to Bryan (2017),

arrowroot starch has more starch than potato which means that fewer amounts of arrowroot

starch can be utilized but it will still have the same effect as the potato starch.

Arrowroot Starch vs. other starches

Arrowroot starch is a healthier option than other starches since it is grain-free which

means it is a big help for people who has a hard time digesting, as said by (Dr. Mercola, 2016).

Arrowroot powder is gaining popularity due to corn allergies and sensitivities or to avoid

anything GMO and pesticide-laden which is cause by cornstarch, another example of a

thickening agent. (Bryan, 2017)

Also according to Bryan last 2017, cornstarch is the most common thickening agent used

in cooking but arrowroot starch can easily replace it. A benefit of arrowroot starch is it has no

taste and it leaves food glossy and clear, whereas cornstarch has a slight taste and leaves food

cloudy and opaque.

These health benefits of arrowroot starch may help in utilizing it as a starch in agar since

it will be used as the culture media for oyster mushroom spawn.

 Methodology

Research Design

In this study, an experiment will been conducted. In the experiment, there are 2 different

set-ups with 3 trials wherein the colonial diameter of the mycelia grown in Arrowroot Dextrose

Agar (Experimental group) and Potato Dextrose Agar (Control group) will be measured to test its

mycelial growth.

Source of Materials

The arrowroot powder and agar powder will be bought online through Shopee, the

potatoes will be bought from Paciano Market, the oyster mushrooms will be bought from Green

Grocer Manila, and the 99% Isopropyl Alcohol and laboratory gloves will be from a medical

supplies store in Parian, Calamba City.

Location of the Study

The whole procedure will be done at the researcher’s residence at Paciano, Calamba City,

Laguna.

Preparing of the Arrowroot Dextrose Agar

The researcher will prepare the arrowroot starch, sugar, agar powder, and other materials

such as the pot where the boiling will occur and pressure cooker. The researcher will then boil

the 15 grams arrowroot powder with 1 liter water then put the 20 grams sugar and 50 grams agar

agar powder with it. After boiling the media it will be divided and put in 3 sterilized petri dishes

and left to cool and dry. The petri dishes will then be subjected to sterilizing by putting it in a

pressure cooker or autoclave for 30 minutes at 121°C and 15 psi. Place the petri dishes over one
another and let the culture media dry in a sterilized container with paper at the cover to make

sure that air doesn’t go inside and contaminate it.

Slice an oyster mushroom in half and get a small tissue from it in the center. Do this process

inside an improvised glove box to avoid contamination from the air and make sure that it doesn’t

make contact with any surface while doing this process. Carefully place the oyster mushroom

tissue inside the culture media then put the petri dishes back in the container with paper at the

cover. Incubate the culture media with the Oyster Mushroom tissue for 2 weeks.

Data Gathering Techniques

Observe the culture media plates after 2 weeks if there is any growth in the petri dishes and

test for its colony diameter then compare it to the Mycelial growth of Oyster Mushroom in

Potato Dextrose Agar.

The study will determine if the arrowroot starch can be used as a starch in Dextrose agar

and if it has any differences with Potato Dextrose Agar. Setup A (experimental group) has the

Arrowroot Dextrose Agar and Setup B (control group) has the Potato Dextrose Agar. These data

will be used to answer the problems.

Research Methodology Flowchart

Preparing and Gathering of Materials

Production of the Arrowroot Dextrose Agar

Setup A Setup B

Arrowroot Dextrose Agar Potato Dextrose Agar

Measuring of the colonial diameter to

determine its mycelial growth

Comparison of the mycelial growth of Arrowroot

Dextrose Agar and the Potato Dextrose Agar
 Bibliography

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