HOLY INFANT ACADEMY

Bonifacio Drive, Ilaya Calapan City Oriental Mindoro 5200

This Capstone Research Project entitled

Anthracnose Inhibiting Activity of San Francisco (Packera franciscana) and Madre De Cacao
(Gliricidia sepium) Leaves Extract on Banana (Musa acuminata) fruit.

In Partial Fulfilment of the Requirements in Capstone Research

Is presented by
Bae, Carl A.
Ante, Ysha Jamilla
Casapao, Viktoria Mae
Coz, Angela Jane Rose
Arellano, Kurt Allen M.

Student Researchers

To
Science and Research Department

With
Mr. Mark Noel C. Leyesa
Research Adviser
April 2020

[AUTHOR NAME] 1
 ACKNOWLEDGEMENT

The researchers would like to express their heartfelt gratitude to all the people who have

supported us which led to the success of this study.

First and foremost, to Almighty God for enlightening the researchers and giving them the

talents, skills, and good health to pursue this Capstone project:

Mr. Mark Noel Leyesa, who served as the adviser and provided oversight and direction to

the researchers, deserves our heartfelt thanks. His persistence and time helped the researchers to

learn more and make steady progress on the study. The related ideas, heartwarming inspirations,

and constant updates kept the researchers on track, and most notably, for being such an

approachable advisor with a notable optimistic mindset.

All Grade 12- St. Blaise who helped and encouraged the researchers in continuing to work

on this project deserves a pat on the back.

And, most notably, the researchers are thankful to the researcher's family for their

encouragement and unending assistance in completing this project.

C.A.B

Y.J.A

A.J.R.C

K.A.A

V.M.C

[AUTHOR NAME] 2
 ABSTRACT

The constant effort of the government to increase food production resulted in the

Utilization of synthetic chemicals. However, continuous reliance on chemicals has depleted the

quality of our agricultural soil. Effects in nutrient content, acidity, and even the presence of

pathogens are some of the impacts to the environment hence, the promotion of organic farming

Thus the researchers conducted a study entitled “Anthracnose-Inhibiting Activity of San

Francisco (Packera franciscana) and Madre de Cacao (Gliricidia sepium) Leaves Extract on

Banana (Musa acuminata) fruit” focusing on the potential fungicidal properties of San Francisco

(Packera franciscana) and Madre de Cacao (Gliricidia sepium) against Anthracnose in banana

(Musa sapientum). Effectiveness was measured in terms of shelf-life and disease severity rate.

Difference levels of concentration were tested: T1-100% San Francisco leaves extract, T2-b. 50%

San Francisco Leaves Extract and 50% Madre de Cacao Leaves Extract, T3- c. 100% Madre

de Cacao Leaves Extract. Ketoconazole was also included as the reference standard. One-Way

ANOVA was utilized to determine the most effective level of concentration.

Analysis of data revealed that Treatment 3: (100% Madre de Cacao Leaves Extract)

showed the longest shelf life of 7.67 days a disease severity rate of 57.64% all recorded in tree

trials.

No significant difference between the best concentrations of Madre de Cacao and San

Francisco leaves extract and the positive control is observed as Ketoconazole exhibited a shelf life

of 7.33 days and an average of disease severity rate of 57.88% which is comparable to Treatment

3.

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 The result of this study indicated that the plant extracts significantly demonstrated the

longest shelf life especially in the second concentration. Thus, Treatment 3 with 100% Madre de

Cacao Leaves extract manifest fungicidal properties. After gathering the observed results the

researchers took into consideration the possible improvements in the study. First would be to

conduct of a fungi bioassay for the experimentation period and phytochemical screening of the

two leaf extracts, secondly would be to employ other extraction methods like the use of methanol,

ethanol or acetone extract the phytochemicals as well as to test the plant extract to other species of

fungi dominant the country. Finally, future researchers are advised to conduct a similar study with

altered variables.

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 Table of Contents

Acknowledgement ………………………………………………………………………… 2

Abstract ……………………………………………………………………………………. 3 - 4

Introduction ………………………………………………………………………………... 7 - 12

Background of the Study…………………………………………………………... 7

Significance of the Study………………………………………………………….. 9

Scope and Delimitation …………………………………………………………… 10

Definition of Terms ……………………………………………………………….. 11 - 12

Review of Related Literature …………………………………………………………….. 13 - 18

Research Methodology………………………………………………………………......... 19 – 21

Research Locale ……………………………………………………………........... 19

Research Design……………………………………………………………........... 19

Sample and Data Collection Techniques …………………………………............. 19

Data Gathering Procedure …………………………………………………........... 21

Statistical Treatment………………………………………………………............. 21

Results and Discussion …………………………………………………………………….22 - 29

Summary…………………………………………………………………………………... 30 - 31

Conclusion…………………………………………………………………………………. 31

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Recommendations ………………………………………………………………………......31

References ……………………………………………………………………………….... 32-33

Appendices ……………………………………………………………………………....... 34-45

Appendix A………………………………………………………………………....34

Appendix B………………………………………………………………………....35

Appendix C………………………………………………………………………....36 - 41

Appendix D………………………………………………………………………....42 - 44

Appendix E………………………………………………………………………....45

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 CHAPTER I

INTRODUCTION

Background of the Study

After having given the privilege to be converted into a component city on Year 1982, the

municipality of Calapan have more capabilities to upgrade, enhance, and enrich its industries in

order to reap the potentials of a port town where goods, passengers, and cargo pass through almost

round the clock. This then results to the need to address and focus on the crops and products trading

issues, considering the damage and spoilage of items being bartered because of a disease (such as

Anthracnose) is inevitable. One of the crops being imported in town are Bananas. However,

mountainous terrains and curved roads make the transportation of crops harder and slower.

Because of this, crops are over-ripened and are affected by diseases such as anthracnose that will

result to bad quality and incapability of being sold and eaten. This study is conducted to find a

viable solution in prolonging the shelf life of the banana fruit by using the extract of Madre de

Cacao and San Francisco. Their leaf has medicarpin and tannin which are anti-fungal compounds

that control the reproduction of fungus Colletotrichum musae, which affects banana fruits. (Sahin

et al, 2004) Madre de Cacao and San Francisco is commonly seen all over Oriental Mindoro.

That is why the researchers had conceived an idea that would be helpful for the lessening

or even the removal of the disease (anthracnose) to the specified product, (which is a fruit called

Banana) that will be used in this study.

Statement of the Problem

Can San Francisco (Packera franciscana) and Madre De Cacao (Gliricidia sepium) Leaves

Extract control the growth of Anthracnose and prolong the shelf-life of Banana Fruit.

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Sub-Problem

1. What is the most effective among the three concentrations of San Francisco and Madre de

Cacao leaves extracts in terms of controlling the growth of anthracnose?

a.100% San Francisco leaves extract

b.50% San Francisco leaves extract and 50% Madre de Cacao leaves extract

c.100% Madre de Cacao leaves extract.

2. What is the best concentration in the different treatment of San Francisco and Madre de

Cacao leaves extract in controlling the growth of Banana Anthracnose in terms of:

a. Shelf-life

b. Disease Severity Rate

3. Is there a significant difference between the effectiveness of the different ‘Concentrations

of San Francisco and Madre de Cacao leaves extract treatment and the commercially

available fungicide agent - Ketoconazole in terms of:

a. Shelf-life

b. Disease Severity Rate

Statement of the Hypotheses

The extract of San Francisco (Packera franciscana) and Madre De Cacao (Gliricidia

sepium) Leaves are effective in controlling the Anthracnose growth and prolonging the shelf-life

of Banana Fruit.

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Sub-Hypotheses

• The most effective concentration in terms of controlling the growth of anthracnose

is Concentration C. Due to the fact that the 100% Madre de Cacao leaves extract are comprised of

blended components containing Tannin and Medicarpin as well as antifungal, anti-amoebic and

anti-cancerous activities which are effective against plant fungi and restricting the growth of

anthracnose.

• The best concentration in controlling the growth of Banana Anthracnose in terms

of shelf life is the 100% Madre de cacao leaves extract is an efficacious concentration that

lengthens the spoilage date of Bananas. And for the Disease Severity Rate is also the 100% Madre

de cacao leaves extract are the efficient disease lessening treatment for the bananas

• There is a significant difference between the effectiveness of the different

Concentrations of San Francisco and Madre de Cacao leaves extract treatment and the

commercially available fungicide agent - Ketoconazole in terms of:

a. Shelf Life

b. Disease Severity Rate

Significance of the Study

In our country, chemical and inorganic fungicides are used in treating certain banana

diseases such as Anthracnose without knowing its consequences to cur nature and to our health

these chemicals can contribute to pollution and harmful results to our environment and to us

humans. Using these two plants can be an effective way to produce an eco-friendly and more

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affordable fungicide agent that is made out of natural and easy-to-get materials. This research is

significant to the following

Farmers and Gardeners. The findings of this study will benefit our farmers in contributing diseases

to crop production. This will help them increase their crop yield without depleting the soil quality

for future use. Local farmers will also be conscious and motivated to re-grow more of these plants

and will also be driven to discover afferent potentials of other botanicals then making them

cultivate more thus, helping nature to re-grow itself once again. Eventually, there will be an

increase in the agricultural profit and better maintained physical health conditions due to the least

intake of harmful vapours from synthetic fungicides.

Agricultural Agencies/Aspects. The result of this study will provide a more accessible solution to

control the growth of Anthracnose in bananas. This will greatly help the agricultural agencies or

aspects for the possible improvement of controlling the growth of Anthracnose. It may also bring

satisfaction for having progressive harvests.

Future Researchers. The result of this study will provide baseline data for future research studies

on the potential fungicidal properties of Madre de Cacao and San Francisco leaf extracts thus, this

will open further tests and studies regarding the potential fungicidal properties of Madre de Cacao

and San Francisco leaf extracts

Scope and Delimitation

This Science Investigatory Project entitled “Anthracnose Inhibiting Activity of San

Francisco (Packera franciscana) and Madre de Cacao (Giiricidia sepium) leaves on Banana (Musa

paradrsiaca) is only limited to studying the potential of Madre de Cacao and San Francisco as an

organic fungicide against banana Anthracnose

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 It is mainly concerned with the screening of the potential fungicidal properties of Madre

de Cacao and San Francisco treat extracts. Moreover, it is limited in prolonging the shelf life and

observing the disease severity after application of the Anthracnose-infected bananas.

The researchers will use banana Anthracnose as the subject of the experiment. Treatments

using Madre de Cacao and San Francisco leaf extracts served as the experimental control. To

compare the potential fungicidal properties, the commercial treatment. Ketoconazole will be use

and 4 negative control will also be prepare, given no treatment for the whole observation period.

The preparation and experimentation will be conducted at the researcher's house.

Definition of Terms

Anthracnose- It refers to the black spots that can be seen in the surface of the banana that happens

to spread swiftly throughout the banana due to the fungus causing it to be diseased and be spoiled.

Anthracnose infected bananas- It refers to those banana that had been invaded and deeply affected

by the anthracnose defect.

Disease Severity Rate- It refers to the rate of how fast does the banana begun affected due to

anthracnose that had spread throughout the shell of it.

Ketoconazole- It is a commercial fungicide that can be applied externally to fight the growth of

fungi (or fungus). Hence, it is used by the researchers to one of their set ups to test its effectivity

along with the other set ups (which were organically made).

Madre De Cacao Leaves- It refers to one of the leaves that were used by the researchers. And its

purpose is to be extracted and mixed in with a specific concentration with the San Francisco

Leaves. So that, the researchers would know whether if they show any significance effectiveness

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in restricting the growth of anthracnose, prolonging its shelf life, and controlling the disease

severity rate of the banana(s).

San Francisco Leaves- It also refers to one of the leaves that were used by the researchers. And

this material were to be extracted and mixed in with a specific concentration with the Madre de

Cacao leaves. And this is to forsee if they show any significance effectiveness in restricting the

growth of anthracnose, prolonging its shelf life, and controlling the disease severity rate of the

banana(s) which will be observed by the researchers.

Shelf Life- It refers on how long does the banana had been prolonged.

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 REVIEW OF RELATED LITERATURE AND STUDIES

Agriculture is one of the primary contributors to the Philippines’ gross domestic product.

It is a source of income and subsistence among many Filipinos. In agriculture, competition is high

on the quality and number of food production. Likewise, there is an increasing demand for food

production for the increasing population in the country. As such, farmers tend to rely on extensive

use of fungicides to increase yield.

Oayan, Franck et al. (2009) states that a Tremendous increase in crops associated with the

‘green’ revolution would not have been achieved without the contribution of these synthetic

compounds, However, concerns have arisen over the potential impact of fungicide to our

environment and the harmful effects it can bring when applied which can potentially decrease crop

production.

Furthermore, the health of most agricultural workers is being compromised. There are

many studies documenting the relationship between pesticide use and its effect on the farmer's

health. Chronic effects of pesticide exposure include cancer in adults and children, adverse

reproductive outcomes, delayed neuropathy, and neurobehavioral effects. People who are directly

exposed to pesticides are the handlers who mix, load, and apply pesticides, and workers cultivating

and harvesting crops. They are at risk of developing acute poisoning or even death from extensive

exposures.

In fact, from 1977 to 1987 pesticide use in our country, including fungicide increased by

325%, however, crops like rice yield increased by only 30% during this time, Fungicide imports

have continued to increase, while rice and corn yields have increased at a much smaller rate

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(Tirado, Reyes and David, Bedoya, 2008). This data sealed that synthetic fungicide and other

pesticides slightly contribute to the increase of crop productions.

Meanwhile, soil degradation is linked to the application of synthetic fungicides and other

chemicals. The N fertilization, the soil moisture, soil temperature, and also the presence of

pesticide residues in the agricultural environment may disturb the natural degradation process.

Some substrates, including pesticide, may inhibit, but also stimulate microbial activity. Thus,

disturbances of the microbial activity may influence the important biogeochemical processes of

the entire soil environment (Stefani, Angelo, et al,, 2012)

Synthetic fungicides can reach surface water through runoff from treated plants and soil,

Contamination of water by pesticides is widespread. More than 90 percent of water and fish

samples from all the streams contained one, or more often, several pesticides and fungicides.

Fungicides were found in all samples from major rivers with mixed agriculture and urban land use

influences and 99 percent of samples of urban streams (Kole et al., 2001)

The main objective of this research is to produce an alternative, affordable and eco-friendly

fungicide that is made out of natural and easy-to-get materials. The proponents utilized San

Francisco and Madre de Cacao leaves as the main materials to produce an organic fungicide. The

leaves are also growing abundantly in the locality making it very convenient to gather and re-create

the organic fungicide.

Banana is undoubtedly the most widely and commonly grown fruit in the Philippines. It

market has wide potentials here and abroad. It is not only a dessert fruit, it can also be made into

banana sauce, fried chips, flour purses, vinegar and wine. It is also used as a substitute for our

staple food (Estrella 2015)

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 Banana (Musa paradisiaca, family Musaceae) is a central fruit crop of the tropical and

subtropical regions of the world’s oldest cultivated crop (Kumar et al., 2012). As a diet, banana is

an affluent source of carbohydrates with calorific value of 67 calories per 100g fruit and is one of

the most well-likes and widely traded fruits across the world (Emagaet al,. 2008; Kumar et al.,

2012)

The Philippine banana industry is one of the country’s best agricultural products. It is

estimated that 35 billion pesos are invested in this agricultural industry which includes the cost of

infrastructure, planting and distribution. However, the productivity of the banana affected by

various diseases. Among these diseases, banana anthracnose which is caused by Colletotrichum

musae is the most common and serious disease.

Anthracnose is a widespread disease of banana fruits. The disease produces unsightly

blemishes on the fruit peel, which makes them unacceptable to consumers. Infection normally

begins as small black circular specks during the pre-harvest stage. Affected fruits ripen

prematurely, rot and eventually shrivel.

The most important disease problem of export bananas (Cavendish cultivars) is crown rot.

This disease may involve several species, but the most commonly associated organism is

Colletotrichum musae (Green & Goos, 1963; Griffee & Burden, 1976; Finlay & Brown 1993). C.

musae also causes anthracnose lesions which commonly appear on the fruit peel after-ripening

(Grifee & Burden, 1974). Control of most banana fruit infections is through the application, shortly

after harvest, of fungicides as a dip or spray (Thompson & Burden, 1995). In the past, the most

commonly used fungicides were benzimidazoles such as benomyl and thiabendazole (TBZ), but

C. musae has developed resistance to these fungicides (Griffee, 1973: Slabaugh & Grove, 1982;

de Lapeyre de Bellaire & Dubois, 1997).

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 Knowing that there are many cases of banana anthracnose and that San Francisco and

Madre de Cacao leaves, which possess the chemical components, fit for the production of an

organic fungicide, gives the researcher an idea to pursue a study on preparing natural fungicide.

San Francisco and Madre de Cacao are plants growing abundantly in many parts of Calapan City,

the City in the province of Oriental Mindoro.

Hajra Faiz, Shazia lram, and Anam Rasool (2016) conducted a study entitled “Management

of Mango Diseases Anthracnose and Blossom Blight by Eco-friendly Methods’” They found out

that eucalyptus extract was effective in controlling diseases and growth of inhibition of

anthracnose. Even though different botanical plant extracts were tested, such as the akk, neem, and

garlic, the eucalyptus showed more potential in controlling the growth of anthracnose. The other

extracts also showed potentials. Thus, akk, neem, garlic, and eucalyptus plant extracts are effective

as organic against anthracnose.

The result of Hajra Faiz, Shazia Iram, and Anam Rasool’s study gave an idea to the

researcher to use San Francisco and Madre de cacao against banana anthracnose since it harbored

the same chemical components that the akk, neem, garlic, and eucalyptus plants all had in common.

According to current resources, a form of Anthracnose disease is infecting a widespread

amount of banana plants in the Philippines, All plants are genetically identical, which prevents the

evolution of disease resistance. Researchers are examining hundreds of wild varieties for

resistance. It has yet to reach the Americas; however, this fungus can travel through the air from

various decaying organic matters. This is how anthracnose travels and it is most likely to travel

into Latin America. The only effective defense is the production of new fungicides.

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 Fungicide, also called antimitotic, any toxic substance used to kill or inhibit the growth of

fungi. Fungicides are generally used to control parasitic fungi that either cause economic damage

to crop or ornamental plants or endanger the health of domestic animals or humans. Most

agricultural and horticultural fungicides are applied as sprays or clouds of dust. Seed fungicides

are applied as a protective covering before germination. Systematic fungicides, or

chemotherapeutants, are applied to plants, where they become distributed throughout the tissue

and act to eradicate existing disease or to protect against possible disease. In human and veterinary

medicine, pharmaceutical fungicides are commonly applied as topical antifungal creams or are

given as oral medications.

Fungicide residues have been found on food for human consumption, mostly from post-

harvest treatments. Some fungicides are dangerous to human health. such as vinclozolin, which

has now become removed from use. Ziram is also a fungicide that is thought to be toxic to humans

if exposed chronically. A number of fungicides are also used in human health care.

In addition, the use of such fungicides may cause adverse effects to terrestrial and aquatic

ecosystems if fungicide residues persist in soil, or if they migrate off-site to surface and ground

waters (Wightwick, Adam, et al., 2008)

Agricultural researchers believed that there are proven alternatives to this expensive and

harmful agriculture system. Farmers are already fertilizing soils and protecting crops with organic

and sustainable techniques that work with nature, not against it, and which can provide food for

all (Pretty and Badgely, 2003).

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 Plants are a good source of antifungal agents. A large number of plant compounds have

been reported to have antifungal activity. Well-known examples are flavonoids. |actones, proteins,

sulphur compounds, cyanogenic glycosides, and essential oils (Sokovic, Marina D. 2013)

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 CHAPTER II

RESEARCH METHODOLOGY

Research Locale

The study was conducted within each of the members’ premises as there is a strict rule of

not going out due to the possible exposure of the COVID 19 virus. However, the experimentation

was done and facilitated at one of the researcher’s house in Canubing I, Calapan City due to the

reason that it is a safe, clean and well-conditioned environment for both the extract making and

the whole 11-day observation. The researchers chose to set a one day schedule for the collection

of materials and the whole experimentation process and for it to be done in the said area

considering that it is secluded and a safe place for conducting the whole experiment. The

researchers also determined that it was the best place to effectively monitor and update the changes

of the bananas without violating any rules and giving the accessibility of the other members via

messages and videos

Research Design

In this study, the experimental research method will be used to utilize in order to determine

the relationship between an independent and another dependent variable in a population.

Experimental method is a problem solving approach that the study is described in the future on

what will happen when variables are controlled and manipulated.

Sample and Data Collection Techniques

Materials and Equipment’s.

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 The researchers gathered all the materials and the major components of the study such as

the San Francisco and Madre de Cacao leaves from Calapan City. The leaves were measured on a

weighing scale of 1 kg. Anthracnose-infected bananas were collected from Calapan Market. The

infected bananas were collected and placed in a container. Commercial fungicide — Ketoconazole

was also provided and bought in Calapan City.

Extraction of leaves

1 kg San Francisco and Madre de Cacao leaves were taken from the Ante's Residence and

washed and air-dried for 30 minutes. They were cut into small pieces, pounded and filtered

thoroughly using the cotton cloth. The resulting product was transferred into the 100 mL spray

container and measured into different concentrations.

Preparation of Extract

After the extraction, the extracts were measured into:

a.100% San Francisco Leaves Extract

b.50% San Francisco Leaves Extract and 50% Madre de Cacao Leaves Extract

c.100% Madre de Cacao Leaves Extract

The prepared extract were placed into 100ml spray bottle and refrigerated to avoid spoilage.

Application of Organic Fungicide

An experiment was conducted to test the effectiveness of the fungicide. Commercially

available fungicide - Ketoconazole was bought in Calapan City and transferred to the 100 ml spray

container. Five set-ups were prepared and labeled set-up A, set-up B, set-up C, set-up D, and set-

up E, The prepared bananas in the container were split into five each set-up. 100 % San Francisco

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leaves extract was applied to set-up A, 50% San Francisco Leaves Extract and 50 % Madre de

Cacao leaves extract was applied to set-up B, the 100 % Madre de Cacao leaves extract was applied

to set-up C and the commercially available fungicide - Ketoconazole was applied to set-up D.

Changes between the five experiments were observed within a week to determine the effectiveness

of the organic fungicide prepared, the average shelf-life of the bananas were recorded in a data

table.

At the same time, the five anthracnose-infected bananas were also used for the examination

of disease severity. For the sake of data collection of banana diseases anthracnose, a per forma

were designed to note down the disease severity. The per forma was filled by rating the severity

of each disease according to a severity scale of 1-5 by the careful observation from each side of

the fruit.

Data Gathering Procedure

Changes within the bananas anthracnose inhibiting activity will be gathered and will be

recorded in a table based on the disease severity before and after the extracts were sprayed in the

specimens.

Statistical Treatment

Results will be recorder, tabulated, and statistically analysed. Analysis of Variance

(ANOVA) will be used to identify the significant difference among the five set-ups.

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 CHAPTER III

RESULTS AND DISCUSSION

Results

The study evaluated the potential fungicidal properties of San Francisco and Madre de

Cacao leaf extract through the shelf-life and the disease severity of the infected bananas. The

fungicidal properties of San Francisco and Madre de Cacao leaf extract was determined based on

the length of time the bananas lasted and the disease severity after the extracts were applied.

TABLE 1
Average Shelf-life and Disease Severity Rate of Anthracnose-Infected Bananas caused by 100% San
Francisco Leaves Extract after exposure

Replicates Average Shelf-life Disease

(Days) Severity

Trial 1 7 54.58%
Trial 2 6 58.33%
Trial 3 6 64.58%

TOTAL 19 177.49%

MEAN 6.33 59.16%

Table 1 represents the average Self-life and Disease severity rate of Anthracnose-infected

bananas caused by San Francisco leaves extract after exposure. Trial 1 has the longest time that

prolonged the shelf-life of the banana with 7 days and has the lowest disease severity rate of

54.58%, followed by trial 2 with the shelf-life of 6 days and Disease severity rate of 58.33% and

last is trial 3 with the shelf-life of 6 days and Disease severity rate of 64.58%.

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 As shown in Table 1, trial 1 is the best replicate in concentration 1 as exhibited the longest

shelf-life and the lowest disease severity rate while Trial 2,3 that shows same result in shelf-life

and are less effective than trial 1.

TABLE 2
Average Shelf-life and Disease Severity Rate of Anthracnose- infected Bananas caused by 50% San
Francisco and 50% Madre de Cacao leaves Extract after exposure

Replicates Average Shelf-life Disease

(Days) Severity

Trial 1 4 75.42%
Trial 2 4 74.17%
Trial 3 7 53.33%

TOTAL 15 202.92%

MEAN 5 67.64%

Table 2 represents the Average Shelf-life and Disease Severity Rate of Anthracnose-

infected Bananas caused by 50% San Francisco and 50% Madre de Cacao leaves Extract after

exposure. Trial 3 has the longest shelf-life of 7 days and the lowest disease severity rate of 53.33%,

followed by trial 2 with the shelf-life of 4 days and Disease severity rate of 74.17% and trial 3

which also has the shelf-life of 4 days and Disease severity rate of 75.42%.

As shown in Table 2, trial 3 is the best replicate in concentration 2 as exhibited the longest

shelf-life and the lowest disease severity rate while Trial 1 shows less effectiveness that the other

two.

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 TABLE 3
Average Shelf-life and Disease Severity Rate of Anthracnose- infected Bananas caused by 100% Madre de
Cacao leaves Extract after exposure

Replicates Average Shelf-life Disease

(Days) Severity

Trial 1 7 58.75%
Trial 2 8 57.08%
Trial 3 8 57.08%

TOTAL 23 172.91%

MEAN 7.67 57.64%

Table 3 displays the Average Shelf-life and Disease Severity Rate of Anthracnose- infected

Bananas caused by 100% Madre de Cacao leaves Extract after exposure. Trial 2 and 3 has the

longest shelf-life of 8 days and lowest Disease Severity rate of 57.08%, followed by trial 1 which

has a shelf-life of 7 days and a 58.75% disease severity rate. Concentration 3 has an average of

7.67 days with a 57.64% disease severity rate. As shown in Table 3, Trial 2 and 3 is the best

replicate in concentration 3 as it exhibited the longest shelf-life and the lowest severity rate while

trail 1 shows less effectiveness than the other two.

TABLE 4
Average Shelf-life and Disease Severity Rate of Anthracnose- infected Bananas caused by 100%
Ketoconazole after exposure

Replicates Average Shelf-life Disease

(Days) Severity

Trial 1 8 58.18%
Trial 2 7 57.73%
Trial 3 7 57.73%

TOTAL 22 173.64%

MEAN 7.33 57.88%

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 Table 4 shows the Average Shelf-life and Disease Severity Rate of Anthracnose- infected

Bananas caused by 100% Ketoconazole after exposure. Trial 1 has the longest shelf-life of 8 days

and have a disease severity rate of 58.18%, followed by trial 2 and 3 which has a shelf-life of 7

days and the lowest disease severity rate of 57.73%. Concentration 4 has an average shelf-life of

7.33 days with a 57.88% disease severity rate. As shown in the Table 4, trial1 is the best replicate

in concentration 4 as it exhibited the longest shelf-life and an average disease severity rate.

TABLE 5
Summary of the Average Shelf-life and Disease Severity Rate of Anthracnose- infected Bananas in the
Three Concentrations and Positive Control after Exposure

Treatment Average Shelf-life Disease

(Days) Severity

Concentration 1 6.33 59.16%

Concentration 2 5 67.64%
Concentration 3 7.67 57.64%
Ketoconazole 7.33 57.88%

Table 5 shows the Summary of the Average Shelf-life and Disease Severity Rate of

Anthracnose- infected Bananas in the Three Concentrations and Positive Control after Exposure.

Concentration 3 has the longest shelf-life of 7.67 days and an average disease severity rate of

57.64%, followed by Ketoconazole which is the positive control with an average shelf-life of 7.33

days and with also a 57.88% disease severity rate. Concentration 1 came next with an average

shelf-life of 6.33 days and has the average disease severity rate of 59.16%. Concentration 2 came

last with the shortest average shelf-life of 5 days and has the highest disease severity rate of 67.64%

[AUTHOR NAME] 25
Discussion

TABLE 6
Summary of the Difference of the three experimental concentrations in terms of
Shelf-life

Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Column 1 3 19 6.333333 0.333333
Column 2 3 15 5 3
Column 3 3 23 7.666667 0.333333

ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 10.66667 2 5.333333 4.363636 0.067622 5.143253
Within Groups 7.333333 6 1.222222

Total 18 8

Table 6 shows the summary of the difference of three experimental concentrations in terms

of shelf-life. Results of ANOVA shows that the three experimental concentrations reveal

significant difference in terms of shelf-life. The computed f-value of 4.36 was lower than the

critical f-value 5.14. Thus, there is no significant difference between the three experimental

concentrations in terms of controlling the growth of Anthracnose in bananas.

TABLE 7
Summary of the Difference of the three experimental concentrations in terms of
Disease Severity Rate

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 Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Column 1 3 177.49 59.16333 25.52083
Column 2 3 202.92 67.64 153.9727
Column 3 3 172.91 57.63667 0.929633

ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 174.2513 2 87.12563 1.448688 0.306668 5.143253
Within Groups 360.8463 6 60.14106

Total 535.0976 8

Table 7 shows that summary of the difference of the three experimental concentrations in

terms of disease severity rate. Result of ANOVA shows that the three experimental

concentrations reveal significant difference in terms of the disease severity rate. The computed f-

value of 1.44 was lower than the critical f-value of 5.14. Thus, there is no significant difference

between the three experimental concentrations in terms of controlling the growth of Anthracnose

in bananas.

TABLE 8
Summary of the Difference of Best Concentration against the Positive Control in terms of
Shelf-life

T-crit Computed Interpretation Decision

t-Value
Concentration 3
vs. Positive 2.13 -0.71 No significant Accepted
Control

[AUTHOR NAME] 27
 Table 8 presents the summary of the difference of the best concentration against the

positive control in terms of the shelf-life of bananas. Results of t-test Concentration 3 and the

Positive control shows a computed t value of -0.71 which is lower than the tabular value of 2.14.

Thus, the null hypothesis is accepted.

TABLE 9
Summary of the Difference of Best Concentration against the Positive Control in terms of
Disease Severity Rate

T-crit Computed Interpretation Decision

t-Value
Concentration 3
vs. Positive 2.14 -0.42 No significant Accepted
Control

Table 9 presents the summary of the difference of the best concentration against the

positive control in terms of the disease severity rate. Results of t-test Concentration 3 and the

Positive control shows a computed t value of -0.42 which is lower than the tabular value of 2.14.

Thus, the null hypothesis is accepted.

According to the Agroforestry Database in 2009 and to De Boer et al, G sepium has been

shown to thave hogh antifungal activity. G sepium contains various phytochemicals like

flavonoids, sterols, alkaloids, glycosides, tannis, saponins, (Sahin et al, 2004) medicarpin,

coumarin, and coumaric acids have beddn isolated and characterized from the plant that showed

antifungal and antibacterial properties.

Phytochemicals such as flavonoids, effectively controls fungal pathogen such as Fusarium

oxysporum. Medicarpin increases the release of phytoalexins which is released by plants to combat

pathogenic infection. Coumarin and coumaric acids both prevent the entrance of nutrients into a

[AUTHOR NAME] 28
fungal cell thus resulting in the fungu’s cell lysis or death. Saponin causes loss of membrane

integrity and is lethal to fungal pathogens.

Several studied also revealed that G. sepium and P. franciscana have medicarpin content

which exhibited potent anti-fungal activity towards Fusarium spore, by inhibiting their

germination and hyphal growth and has cytotoxic abilities. Coumarins, especially esculetin, inhibit

cell growth and cell cycle progression by including arrest of the G phase in cells. Coumaric acid,

on the other hand, shows strong antioxidant properties.

[AUTHOR NAME] 29
 CHAPTER IV

SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

Summary

To assess the potential fungicidal properties of San Francisco and Madre de Cacao leaf

extracts, the shelf-life and disease severity life of the infected bananas were used. Thus, the

fungicidal properties of San Francisco and Madre de Cacao leaf extracts were determined based

on the length of time the bananas lasted and the disease severity after the extracts were applied.

As the experimentation was commenced and came to an end with the span of 11 days, the

researchers were able to gather data and calculate the mean number of shelf-life and diseases

severity rate to determine the effectivity of the treatments per banana piece. The salient summary

yielded results, wherein different concentrations of San Francisco and Madre de Cacao leaves

extract influence fungi development, extending the shelf life of Anthracnose-infected bananas and

controlling the growth of anthracnose present in each banana pieces, as observed with their disease

severity rate. The most efficacious among the three extracted concentrations was the Treatment 3

(100% Madre de Cacao leaves Extract) as it exhibited an average shelf-life of 7.67 and 57.64%

disease severity rate. In terms of shelf-life with 7.33 days and disease severity rate of 57.88 percent

Ketoconazole, the commercially available fungicide did not show the strongest fungicidal

properties of all in controlling the growth of banana Anthracnose. This shows that Treatment 3 is

commensurable to the positive control of Ketoconazole since their fungicidal properties are

relatively close to each other. With reference to the growth of banana Anthracnose in terms of the

shelf-life and disease severity rate, the experimentation yielded results in which, proves that there

is a significant difference between the three experimental concentrations. Thus, concluding that

there is no significant difference between the best concentration of San Francisco and Madre de

[AUTHOR NAME] 30
Cacao leaves extract and the positive control in controlling the growth of banana Anthracnose in

terms of the shelf-life and disease severity rate.

Conclusion

Based on the finding of the study, the researchers concluded that the chemical

composition present in the leaves or plant makes the extract lethal to some extent against banana

Anthracnose. And in terms of shelf-life and disease severity rate, the San Francisco and Madre de

Cacao extracts displayed a great role in controlling the growth of banana Anthracnose. Each

concentration gave different significant effects in controlling the growth of banana Anthracnose

in terms of shelf-life and disease severity rate as a significant difference between the three

experimental concentrations in controlling the growth of banana Anthracnose in terms of the shelf-

life and disease severity rate was found. Meanwhile, there was no significant difference between

the best concentrations of San Francisco, Madre de Cacao leaves extract and the positive control

in controlling the growth of banana Anthracnose in terms of the shelf-life and disease severity rate.

Recommendation

For future use and to give further insights, the researchers suggest the conduct of a fungi

bioassay for the experimentation period and phytochemical screening of the two leaf extracts if it

is possible to attain more concise results. The future researchers may also employ other extraction

methods life the use of methanol, ethanol or acetone extract the phytochemicals. Doing so would

help extract active ingredients of the plants. Testing the plant extract to other species of fungi

dominant the country would also be a suggestion that they could reconsider. And finally, for future

direction for this research, future researchers are advised to conduct a similar study with altered

variables.

[AUTHOR NAME] 31
 REFERENCES

 Department of Agriculture and Fisheries (2016) Anthracnose. Retrieved from

https://www.daf.qld.gov.au/business-priorities/agriculture/plants/fruit-vegetable/diseases-

disorders/anthracnose

 Diamante, J., Dacillo, V.J., Pentinio, J.C. (2017) Oregano and Kamantigue leaf extracts as

a Potential Organic Fungicide against Fusarium oxysporum causing Fusarium wilt in

Banana. San Teodoro National High School.

 Diaz, J. (2017) Anthracnose Inhibiting Activity of Madre de cacao leaf extra.

https://prezi.com/_sd9ohkuyozi/anthracnose-inhibiting-activity-of-madre-de-cacao-leaf-

extra/

 Kuo, P.C. (2015) Analysis of Antifungal Components in the Galls of Melaphis chinensis

and Their Effects on Control of Anthracnose Disease of Chinese Cabbage Caused by

Colletotrichum higginsianum. Retrieved from

https://www.hindawi.com/journals/jchem/2015/850103/

 Nataraj S.K., Munikirishbnappa P.M., Sy C., Reddy B.S., (2017) Codiaeum Variegatum

(Family: Euphoribiaceae). Commonly used Ornamental Plants. (251-255). Kruger Brentt

https://www.researchgate.net/profile/Nataraj_S_K/publication/343306517_Codiem_verig

atum/links/5f22900c299bf13404927ee8/Codiem-verigatum.pdf

 Nattawut, R. (2016, June) Isolation and evaluation of biocontrol agents in controlling

anthracnose disease of mango in Thailand. Retrieved from

https://worldwidescience.org/topicpages/a/anthracnose+disease+control.html

[AUTHOR NAME] 32
 Posada, E.R., (2017) Anthracnose Inhibiting Activity of Madre de Cacao (Gliricidia

sepium) Leaf Extract on Post – harvest Tomato (Solanum lycopersicum). http://sip-

mnhs.blogspot.com/2016/03/anthracnose-inhibiting-activity-of.html

 Roldan, G. (2019) Anthracnose Restraining Property of Madre de Cacao (Gliricidia

sepium) Leaf Extract on Banana (Musa acuminata) fruit The Problem and Its Background.

https://www.academia.edu/28761392/Anthracnose_Restraining_Property_of_Madre_de_

Cacao_Gliricidia_sepium_Leaf_Extract_on_Banana_Musa_acuminata_fruit_The_Proble

m_and_Its_Background

 Supetran, B.L. (2018, June 3) Calapan: A booming port city.BM. Retrieved from

https://businessmirror.com.ph/2018/06/03/calapan-a-booming-port-city/

 Torres, M.L.(2018) MADRE DE CACAO (GLIRICIDIA SEPIUM) AND SINTA

(ANDROGRAPHIS PANICULATA) LEAVES EXTRACT AS BOTANICAL ANIMAL

LICE AND TICKS REMOVER. Journal of Fundamental and Applied Science ISSN 1112-

9867

[AUTHOR NAME] 33
 APPENDIX A

Materials and Equipment’s

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 APPENDIX B

Procurement of Product

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 APPENDIX C

Experimentation

Day 0

Day 1

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Day 2

Day 3

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Day 4

Day 5

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Day 6

Day 7

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Day 8

Day 9

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Day 10

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 APPENDIX D

STATISTICAL COMPUTATION

A. SHELF LIFE

Concentration 1 vs. Ketoconazole

t-Test: Two-Sample Assuming Equal Variances

Variable Variable
1 2
Mean 6.333333 7.333333
Variance 0.333333 0.333333
Observations 3 3
Pooled Variance 0.333333
Hypothesized Mean
Difference 0
df 4
t Stat -2.12132
P(T<=t) one-tail 0.050596
t Critical one-tail 2.131847
P(T<=t) two-tail 0.101192
t Critical two-tail 2.776445

Concentration 2 vs. Ketoconazole

t-Test: Two-Sample Assuming Equal Variances

Variable Variable
1 2
Mean 7.333333 5
Variance 0.333333 3
Observations 3 3
Pooled Variance 1.666667
Hypothesized Mean
Difference 0
df 4
t Stat 2.213594
P(T<=t) one-tail 0.04563
t Critical one-tail 2.131847
P(T<=t) two-tail 0.09126
t Critical two-tail 2.776445

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 Concentration 3 vs. Ketoconazole

t-Test: Two-Sample Assuming Equal Variances

Variable Variable
1 2
Mean 7.333333 7.666667
Variance 0.333333 0.333333
Observations 3 3
Pooled Variance 0.333333
Hypothesized Mean
Difference 0
df 4
t Stat -0.70711
P(T<=t) one-tail 0.259259
t Critical one-tail 2.131847
P(T<=t) two-tail 0.518519
t Critical two-tail 2.776445

B. DISEASE SEVERITY RATE

Concentration 1 vs. Ketoconazole

t-Test: Two-Sample Assuming Equal Variances

Variable Variable
1 2
Mean 59.16333 57.73
Variance 25.52083 #DIV/0!
Observations 3 1
Pooled Variance 25.52083
Hypothesized Mean
Difference 0
df 2
t Stat 0.245714
P(T<=t) one-tail 0.414409
t Critical one-tail 2.919986
P(T<=t) two-tail 0.828818
t Critical two-tail 4.302653

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Concentration 2 vs. Ketoconazole

t-Test: Two-Sample Assuming Equal Variances

Variable Variable
1 2
Mean 57.88 67.64
Variance 0.0675 153.9727
Observations 3 3
Pooled Variance 77.0201
Hypothesized Mean
Difference 0
df 4
t Stat -1.36205
P(T<=t) one-tail 0.12242
t Critical one-tail 2.131847
P(T<=t) two-tail 0.24484
t Critical two-tail 2.776445

Concentration 3 vs. Ketoconazole

t-Test: Two-Sample Assuming Equal Variances

Variable Variable
1 2
Mean 57.63667 57.88
Variance 0.929633 0.0675
Observations 3 3
Pooled Variance 0.498567
Hypothesized Mean
Difference 0
df 4
t Stat -0.42207
P(T<=t) one-tail 0.347335
t Critical one-tail 2.131847
P(T<=t) two-tail 0.694671
t Critical two-tail 2.776445

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 APPENDIX E

FLOW CHART

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