In Vitro Antifungal Activity of Red Onion (Allium cepa) Peel Extract

Against Sporisorium scitamineum

An Official Entry to the School-based Research Fair

Science Investigatory Project

Life Science Category

Glaizelle T. Leonoras

Diana Jean A. Sanico

Research Advisers

Jomar L. Cera

Czarrina Kyle B. Descalso

Myna Patricia B. Gabuya

Researchers

Negros Occidental National Science High School

Victorias City, Negros Occidental

August 2019
 ACKNOWLEDGMENTS

The researchers would like to extend their heartfelt gratitude to everyone that helped

them from the beginning until the end of their research undertaking.

To the God Almighty, all of these would not be possible without His blessing and

guidance. For providing the researchers' strength and patience as they go along their research.

For blessing wisdom to make substantial and good judgments and the courage to remain intact

despite the challenges ahead of them. Through His grace and mercy, this study became a

successful one.

To the parents of the researchers, Mrs. Jonita L. Cera, Mr. and Mrs. Joebert Descalso, and

Mr. and Mrs. Myniel B. Gabuya for giving their love, understanding, and undying support to the

researchers in accomplishing their study.

To Mr. Joebert Ramos, for giving directions, assistance, and lending the expertise of his

chosen field for the completion of the study of the researchers.

To the research advisers, Ms. Glaizelle T. Leonoras, Mrs. Diana Jean A. Sanico and Mrs.

Kalvin Joy Bauno for their time, support, understanding, and patience in answering all of our

questions as well as sharing their knowledge in the field of research.

To Ms. Herian Marie Ibañez, Sir Dexter Balboa, Ms. Niña Janessa Eduardo, Ms. Krystin

Pearl Navarro, and Ms. Necher Abelada for the guidance they gave during the conduct of our

study. To Herbanext Laboratories Inc., for allowing us to utilize their materials and facilities to

complete our study.

i
 To the statisticians, Ms. Ruth Fontanilla and Mr. Bernard Belleza for helping us with

computing our data and interpret our results.

ii
 ABSTRACT

                The sugarcane industry had been the lifeline of Negros Occidental economy. But
commercial varieties of this ratoon crop had been laid unavailable due to Ustilago scitaminea
Sydow or the sugarcane whip smut. Planted smut resistant sugarcanes eventually succumbed to
the disease. Sporisorium scitamineum is the causative agent of the smut disease. Farmers relied
to the use of fungicides. Yet, fungicide immunity became evident. This study aimed to determine
the antifungal property of red onion (Allium cepa) peel extract against S. scitamineum as a
contribution of using phytomedicine against phytopathogen. Extraction was carried out via
rotary evaporation using ethanol as solvent in Herbanext Laboratories. The result showed that
the extract was more effective compared with the marketed fungicides. It was discovered that the
higher concentration the red onion peel was the greater its ability to limit the growth of fungi.

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 TABLE OF CONTENTS

INTRODUCTION
Background of the Study ………………………………………………………. 1

Statement of the Problem ………………………………………………...…….. 3

Hypothesis …………………………………………………………………….…3

Significance of the Study …………………………………………………..…… 3

Scope and Delimitation ……………………………………………………….…..3

METHODOLOGY
Flow Chart ……………………………………………………………………….4

Research Design …………………………………………………………...…….5

Procedure …………………………………………………………...……………5

Data Collection …………………………………………………………….….. 10

Data Analysis …………………………………………………………..……… 11

RESULTS AND DISCUSSION

Findings …………………………………………………………………..…… 12

Summary …………………………………………………………………..…... 16

Conclusion ……………………………………………..……………………… 17

Implication ………………………………………..…………………………….17

Recommendation ……………………………………………………………….18

REFERENCES …………………………………………………………………………19

APPENDICES
Statistical Computation …………………………………………………………22

Documentation ………………………………………………………………… 25

Letters and Certificates ………………………………………………………… 28

 LIST OF TABLES

Table 1. Phytochemical Screening of Red Onion Peel Extract …………………..….13

Table 2. Composition of Commercialized Fungicide Benomyl 50WP ……………....13

Table 3. Radial Growth after the Introduction of Red Onion Peel Extract …………..14

Table 4a. Significant Differences Among Treatments …………………………….…15

Table 4b. Post Hoc Analysis in Scheffe ……………………………………………...15

Table 5. Correlation between the concentration of red onion peel extract and

Radial growth ………………………………………………………….16

 INTRODUCTION

Sugarcane (Saccharum spp.) is one of the world’s largest crops, cultivated in about 26.9

million hectares in more than 109 countries and takes over 60% of the world’s sugar (Dotaniya et

al., 2016). Meanwhile, in the Philippines, Western Visayas plays a big part in the sugarcane

industry and remained as one of the top producers of sugar throughout the years. As of 2015, the

region produced 13.42 million metric tons denoting 58.5% of the national sugarcane production.

Sugar-making has served as the lifeline of the Negros Occidental's economy contributing to more

than half of the country's sugar, making itself the "Sugarbowl of the Philippines." Negros

Occidental is highly respected in terms of exporting sugar that it was able to supply not only

national consumers but international ones as well.

The Ustilago scitaminea Sydow or sugarcane whip smut is a plant disease that emerged in

the Philippines in 1920 and can infect 50-75% of the sugarcane stalk (Luzaran et al., 2012). The

smut infection easily reduced yield by 50% and made ratoon crops unprofitable. Due to its high

infectivity, developed countries have been unable to stay smut free even with the use of

appropriate quarantine measures (Esh et al., 2018).

Sporisorium scitamineum, an airborne pathogenic spore that enters through the buds

(germinating sugarcane shoots) is the causative agent of the sugarcane whip smut. It develops

with the apical and lateral meristems of the plant and induces the creation of a dusty, whip-like,

black structure which is a combination of the plant and fungal tissues. The teliospores of S.

scitamineum can contaminate both soil and water irrigation that may enter other plant paddies

(Wada et al., 1999).

The Philippines had suffered serious economic losses from the sugarcane whip smut.

According to the Bureau of Agriculture in 2012, the sugarcane whip smut was considered as one

1
of the most destructive sugarcane diseases. The presence of the disease was unnoticeable until a

new set of sugarcane has been cultivated. Although it was also widespread to other sugar-

producing countries, the highest genetic diversity of the phytopathogen was found here in the

country. (Yohannes et al., 2011).

The exploration of phytomedicine began due to fungicide immunity. As per the

Philippine Sugar Research Institute (2002), due to the Philippines' climate, commercial varieties

of sugarcane that were supposed to be resistant succumb to the disease.

Onion (Allium cepa) is a well-known crop that is used in culinary arts and traditional

medicine containing polyphenols which are bioactive substances that have multiple biological

properties, such as antioxidant, antimutagenic, antibacterial, antiviral and anti-inflammatory

activities (Vlase et al., 2013), and is widely cultivated in many parts of the world.

The onion peel was often discarded because it can't be used in cooking. However, it was

discovered to contain antioxidants. The outer layer of red onion variety presented high levels of

flavonols (Benmalek et al., 2013). Dimethyl-trisulfide and methyl-propyl-trisulfide were

identified in the essential oil of A. cepa and also displayed fungicidal effects on Penicillum

chrysogenum, Aspergillus niger and species of the respective genera (Pejin, et al., 2016). These

phenolic compounds constituted heavily on how onion peel can exhibit antifungal property. 

Thus, the researchers believed that the red onion peel could be considered as a potential

phytomedicine against phytopathogens.

This study aims to determine the antifungal activity of red onion (Allium cepa) peel

extract against Sporisorium scitamineum. Specifically, it aimed to: (1) Determine the present

phytochemical components in the red onion peel. (2) Determine the radial growth produced after

the introduction of red onion (Allium cepa) peel extract at varying concentrations and

2
commercialized fungicide against Sporisorium scitamineum: a. 30% concentration of red onion

(Allium cepa) peel extract b. 20% concentration of red onion (Allium cepa) peel extract c. 10%

concentration of red onion (Allium cepa) peel extract d. 50 mg of CAT Benomyl 50WP (Positive

control). (3) Determine the significant difference in the radial growth between the

commercialized fungicide and each red onion (Allium cepa) peel extract. (4) Determine the

significant relationship between the different concentrations of red onion (Allium cepa) peel

extract and the radial growth produced.

There was a significant difference after introducing each concentration of red onion peel

extract and the commercialized fungicide. They also wanted to see if there was a significant

relationship among the concentrations of red onion peel extract.

The findings of the study would benefit farmers by considering the use of phytomedicine

against plant pathogens since fungi had developed resistance to fungicidal mechanisms from

marketed fungicides. This study would also give an advantage for the milling companies. It

would help the improvement of crop yield. Thus, there would be an increased rate of sugar

production to export. To the future researchers, the study could play a big part in providing more

information regarding the studies for phytomedicine. Using this, future researchers could

innovate and make a better study to address other problems.

The confines of other in vitro studies were also applied here in the same manner. This

study only focuses on the effect of the red onion A. cepa peel extract on the Sporisorium

scitamineum. The extract would not be applied to the infected sugarcane. The controlled setting

in the laboratory minimized the extraneous factors. Thus, the researchers greatly accentuated that

the alterations during the experimentation on S. scitamineum do not mean that it will yield the

same effects if introduced to the outside environment.

3
 METHODOLOGY

Collection and Extraction of Red Onion

Peel

Collection of Infected Sugarcane

Isolation, Incubation, and Identification

of Sporisorium scitamineum

Microscopy

Phytochemical Screening of Red

Onion Peel

Antifungal Assay

Data Gathering

Disposal

Statistical Analysis

4
Research Design

The researchers followed the Time-series experimental research design. The three red

onion peel treatments were compared to two control groups, the positive control (CAT Benomyl

50WP) and negative control (no treatment application). The control groups and the interventions

were subjected to daily mensuration during the five-day incubation of the Sporisorium

scitamineum. The experiment was done in a controlled environment.

Procedures

Collection and Extraction of Red Onion Peel

The red onion (Allium cepa) peels were collected from E.B. Magalona public wet market.

The samples were cleaned with water and air-dried. The extract was obtained using solvent

extraction method in Herbanext Laboratories, Taloc, Bago City. Four hundred forty-four grams

(444 g) of red onion skin was soaked in one and seventy-eight liters (1. 78L) of 80% alcohol

(ethanol). It was set aside for 48 hours inside a clear glass bottle. The filtrate was sieved and then

transferred into a round-shape rotary flask. The red onion peel extract was concentrated under

reduced pressure in a rotary evaporator at 60 °C at 80 (revolutions per minute) rpm. The

researchers were assisted by a resident chemist to indicate the removal of all present ethanol. The

researchers gained 141 ml of red onion peel extract. After evaporation, the extract was stored in a

refrigerator until use.

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Collection of Infected Sugarcane

The infected sugarcane stalks were collected in a sugarcane plantation in Brgy. Tabigue,

E.B. Magalona. The sugarcanes were physically identified through a growing whip-like, dusty,

curved black shoot that may be several feet in length. The shoot was devoid of leaves and was

slender, flexible and covered by a silvery, thin membrane. Ten infected sugarcane stalks were

collected, parts belonging 15 cm from tip and below of whips was cut and removed. The central

part of the whips was placed inside a sterilized glass bottle and was transported to the laboratory.

Phytochemical Screening of Red Onion Peel

           The extract was subjected to various standardized tests to determine the presence of the

following metabolites: flavonoids, tannins, saponins, proteins, and amino acids.

Test for Flavonoids

Two phytochemical tests were used to determine if there where flavonoids present in the

red onion peel extract. First, lead acetate test was, 0.2 ml of the extract was transferred to a

sterile test tube using a pipette and combined with 0.2 ml of 10% lead acetate. The mixture was

shaken gently to avoid emulsion. A creamy white precipitate would indicate the presence of

flavonoids. In the sodium hydroxide test, 0.2 ml of dilute NaOH was added to 0.2 ml of the red

onion peel extract and was also shaken gently. The extract would appear as intense yellow or a

yellow precipitate would emerge because of the flavonoids.

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Test for Tannins

  One ml of the red onion peel extract was transferred into a test tube. Then, three drops of

neutral 10% Ferric chloride solution were added. A green-black precipitate would indicate the

presence of tannins.

Test for Saponins

          The red onion peel extract was tested by frothing in aqueous solution, 0.5 ml of the red

onion peel extract was transferred using a pipette. The test tube containing the extract was filled

with 2 ml of distilled water. The solution was shaken. Continuous frothing would indicate the

presence of saponins.

Test for Proteins

Biuret test was used where 1 ml of the red onion peel extract was transferred to clean test

tubes. Two ml of 10% NaOH solution and 2 drops of 0.1% Copper sulfate solution were added

to the sample. The presence of proteins would be indicated through the violet or pink color

formation in the sample.

Test for Amino Acids

           For the test of amino acids, the ninhydrin test was used where 1 ml of the red onion

ethanolic extract was transferred to a sterile test tube. One ml of 0.2% ninhydrin solution was

added to the sample then boiled for 8 minutes. The formation of purple color in the sample

would indicate the presence of amino acids in the extract.

7
Isolation, Identification, and Incubation, of Sporisorium scitamineum

The whips were scraped with a sterile spoon to obtain the black (mature) teliospores and

waste materials were removed. The teliospores were disinfected in distilled water containing 3

mg L-1 streptomycin sulfate. Then, it was placed inside three Eppendorf tubes and had undergone

centrifugation. Before desiccation, it was weighed on an analytical balance to find its initial mass

(43 107. 06 mg). The teliospores were desiccated in a desiccator until observably dry. Still using

an analytical balance, the final mass of the black teliospores were taken (356. 84 mg).

For the germination, the desiccated teliospores were inoculated using a sterile inoculation

loop on two SDA (Sabouraud Dextrose Agar) plates. The media were incubated in the dark for

12 hours at 25°C. After incubation, it was transferred into two fresher SDA supplemented with

100ppm chloramphenicol to address potential bacterial contamination. It was again incubated in

the dark for 24 hours at the same temperature.

Microscopy

The sample collected had undergone conventional microscopy to determine if the fungi

was accurate. The teliospores were streaked on two glass slides containing a drop of saline

solution and were heat treated. The microbiologist examined it under a Euromex CKL No. 4

Professional Microscope with 40x magnification. The spores were described as small blackish-

brown and were 5.5-7.5 µm in diameter. After the ten days of incubation, the growth pattern and

characteristics of the fungi were observed. It formed white mats on the SDA culture media. A

small square of whitish mycelium was cut and subjected to another microscopy. The part was

stained with crystal violet. The mycelia were septate and the sporidia were ovoid.

8
Preparation for Different Treatments and Media

The specific quantities of the red onion peel extract were added using a sterilized pipette in

their respected Petri dishes. Then, the SDA media were poured on the dishes. The plates were

gently mixed in orbicular motion to achieve an even distribution between the red onion peel

extract and SDA culture media. It was left for 30 minutes to solidify. Each treatment had three

replicates, along with the positive control (CAT Benomyl 50WP) and the negative control (No

treatment). The concentrations were converted into the following milliliters:

1. Treatment 1: 30 % Red onion peel extract concentration (with 6:14 ml, peel extract to

SDA culture medium ratio)

2. Treatment 2: 20 % Red onion peel extract concentration (with 4:16 ml, peel extract to

SDA culture medium ratio)

3. Treatment 3: 10% Red onion peel extract concentration (with 2:18 ml, peel extract to

SDA culture medium ratio)

4. The positive control group (50 mg CAT Benomyl 50WP to 20 ml SDA culture medium)

5. The negative control group (without the inhibition and CAT Benomyl 50WP)

Antifungal Assay

The antifungal activity of the red onion peel extract was tested using the Poisoned Food

technique in Herbanext Laboratory. Each red onion peel extract concentration was tested in

triplicates. The fungicide (CAT Benomyl 50WP) was dissolved in three SDA plates before it

9
solidified. The agar disc containing the pure isolated S. scitamineum fungi was cut out using

needles. The needles were heat-treated in an alcohol lamp until the needles were red. The cut-out

agar was transferred aseptically to the center of Petri dishes, ensuring that the film-like white

mycelium was touching the agar base. The three treatments, positive control, and negative

control were incubated in the dark at room temperature.

Data Gathering

The efficacy of the red A. cepa peel extract was tested and measured in Herbanext

Laboratory. In the first 24 hours of incubation, the colony growth radius of all groups was

measured by a digital Vernier caliper. The mensuration was repeated when the incubation period

reached 48 hours (2 days), 72 hours (3 days), 96 hours (4 days) and, finally, 120 hours (5 days).

Disposal

All substances and samples had undergone the proper disposal procedure. The Petri

dishes and materials used in the experiment were decontaminated in an autoclave at 121°C for 20

minutes. Then, the agar was scraped off and put inside an autoclavable plastic and sealed again

in yellow plastic with a biohazard symbol. The Petri dishes were washed with Lysol I.C and

bleached and again underwent the autoclave for sterilization. The excess samples used was

placed in trash bins designated for organic wastes.

10
Data Analysis

The data obtained from the tests were subjected to the following descriptive and

inferential statistical treatments. The researchers utilized SPSS Statistics software and used one-

way analysis of variance (ANOVA) to identify whether there was a significant difference in the

radial growth between the commercialized fungicide and each red onion (A. cepa) peel extract.

Post Hoc Analysis was used to linearly compare the difference of all treatments, positive

control and negative control, selecting the day five (5) in the radial growth the experiment.

Pearson Correlation was used to know the significant relationship of the interventions

with the radial growth.

11
 RESULTS AND DISCUSSIONS

Table 1
Phytochemical Screening of Red Onion Peel Extract

QUALITATIVE PHYTOCHEMICAL FOR CONSTITUENTS

Sample Flavonoids Saponins Amino Acids Proteins Tannin

Red Onion
Peel Ethanolic
Extract + + + - +

Note:
Result are specifically related to samples as received.
(-) may denote compound to be present in trace amount.

The results showed the confirmation of the present metabolites in the red onion extract:

flavonoids, saponins, amino acids and tannins. The proteins were assumed to be present in trace

amount in the red onion A. cepa peel extract. Compared to the edible inner part of the red onion,

the phenolic compounds that the red onion skin contained were three to five times higher; it can

greatly inhibit the growth of microorganisms (Chiew, 2014).

Table 2
Composition of Commercialized Fungicide Benomyl 50WP
Active Ingredient:
Benomyl 500g/kg
Inert Ingredients 500g/kg

Table 2 showed the ingredients of Benomyl 50 WP, commercial fungicide used as

positive control. The treatment of sugarcane cuttings with fungicides was difficult to assess

because it only ensures the seed cane was smut free (Bailey, 1979).

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Table 3
Radial Growth after the Introduction of Red Onion Peel Extract
Radial Growth (mm) Averag
N e
Std.
Treatment (per Radial
Day 1 Day 2 Day 3 Day 4 Day 5 Deviation
pay) growth
per day
30%
3 NVG NVG 7.06 10.00 10.96 2.19 .304
concentration
20%
3 NVG 7.44 9.31 10.51 14.60 2.92 .945
concentration
10%
3 7.39 8.46 18.54 31.50 38.14 7.63 .580
concentration
50mg
Commercial
3 NVG 8.61 9.31 13.67 18.31 3.66 .078
fungicide (CAT
Benomyl 50WP)
No Treatment 3 26.39 32.67 45.69 65.89 78.60 15.72 .311

Note: No Visible Growth (NVG)

Table 3 showed that the average daily radial growth of Sporisorium scitamineum was

highest when not treated , whereas it was at its lowest when introduced

with 30% concentration of red onion (A. cepa) peel extract . The

average growth of the control was 15.72 mm per day. Whereas in the positive control (CAT

Benomyl 50WP), the average radial growth daily was 3.66 mm. It could be inferred that the

commercialized fungicide had effectively hindered the phytopathogen based on the results of the

non-treated media. The red onion peel extract (OPE) treatments with 10%, 20% 30%

concentration produced an average radial growth per day, respectively, 7.63mm, 2.92 mm, 2.19

mm. The 30% concentration of red OPE had impeded the growth for two days. While, both the

20% concentration of red OPE and fungicide, had delayed it for a day. Compared with the

positive control, the 30% concentration of red onion peel extract, of red OPE concentration, had

proven to be more effective in the inhibition of radial growth.

13
 According to Franco, et al (2018), the red onion peel exhibited 100% efficacy against the

radial growth of Fusarium moniliforme. Different Allium spp. had also displayed high inhibitory

effects on other radially growing fungi such as Pencillium oxalicum, Fusarium oxysporum,

Botryodiplodia theobromae Macrophomina phaseolina, Aspergillus niger, Rhizopus stolonifer,

and Fusarium solani (Okigbo, et al., 2019).

Table 4.a
Sum of Df Mean F Sig
Squares Square
Between Groups 9410.433 4 2352.608 329.718 .000
Within Groups 71.352 10 7.135
Total 9471.786 14
Significant Difference Among Treatments

This table showed that there was a significant difference between and among the radial

growth of Sporisorium scitamineum under different treatments, , .

Table 4.b
Post Hoc Analysis in Scheffe
Dependent Variable: Radial Growth in Day 5

Linear Comparison Mean Difference Sig.

30% Extract - 20% Extract -3.65 .610

30% Extract - + Control -7.53 .073
30% Extract - 10% Extract -27.18* .000
30% Extract - - Control -67.64* .000
20% Extract - + Control -3.89 .555
20% Extract - 10% Extract -23.54* .000
20% Extract - - Control -63.99* .000
+ Control - 10% Extract -19.65* .000
+ Control - - Control -60.11* .000
10% Extract - - Control -40.46* .000
Note: * The mean difference is significant at 5%.

14
 Table 4.b showed that there was a significant difference between radial growth when treated

using 30% concentration of red onion extract and using 10% concentration of red onion peel

extract. . Between the 30% and the 20 % concentration of red onion

peel were more effective than the 10%.

Table 5
Concentration of
red onion peel N Mean Std. Deviation
extract
30% Extract 3 10.96 1.52
20% Extract 3 14.60 4.73
10% Extract 3 38.14 2.90
Correlation N P
-1.000* 9 .000
Correlation between the concentration of red onion peel extract and radial growth
Note: *Correlation is significant at the 0.01 level (2-tailed). The correlation coefficient is
interpreted according to the scale (Mukaka, MM, 2003): .90-1.0 Very High, .70-.90
High, .50-.70 Moderate, .30-.50 Low, .00-.30 Negligible Correlation.

Table 5 showed that there was a negative correlation between the concentration of red

onion extract and radial growth Sporisorium scitamineum

This means that the less the concentrated the treatment, the greater is the radial growth.

Likewise, the more concentrated is the treatment, the lesser is the radial growth.

There was also a general increase of inhibition when extract concentration was increased

in the study of Okigbo in 2019.

15
SUMMARY

The red onion Allium cepa peel extract had exhibited comparable result to the

commercial fungicide. The growth of the fungi under 30% concentration had been inhibited for

two days. While in for the CAT Benomyl 50WP the growth was impeded for one day.

There is negative correlation between the extracts and radial growth which means the

more concentrated the red onion peel extract was, the more difficult for the fungi to grow.

CONCLUSION

Based on the findings of the study, Red Onion (Allium cepa) peel extract was effective on

the radial growth of Sporisorium scitamineum which causes the sugarcane whip smut disease.

After the introduction of the varying extract concentrations, the radial growth is observably small

compared to the negative control. The average growth per day of the 30%, 20% and 10%

concentration were 7.63mm, 2.92 mm, 2.19 mm in contrast to negative control which was 15.72

mm

In the phytochemical screening, four out of five metabolites were found to be present on

the red onion A. cepa peel extract. Proteins tested negative while the rest tested positive namely:

Flavonoids, Tannins, Saponins, and Amino Acids. Conversely, the components present in the

fungicide was Benomyl (500 g/kg) and inert ingredients (500 g/kg). 

The researchers determined that there is a significant difference between the radial

growth of the fungi using each extract concentration and the usage of commercialized fungicide

using one-way ANOVA. As the concentration increases, the radial growth decreases. After five

days of observation, the formed growth per day was measurably small in diameter (mm), 10%,

20% 30% concentration produced were 7.63mm, 2.92 mm, 2.19 mm respectively. Post Hoc

16
Analysis in Scheffe was used to compare the inhibition with each other, the treatments with the

fungicide, and with the negative control.

Finally, with the use of Pearson Correlation, the researchers discovered that there was a

significant relationship between the concentrations of the Red Onion peel extract and radial

growth of Sporisoriu scitamineum.

To conclude, this study is a method to consider utilizing phytomedicine to eliminate

phytopathogens.

IMPLICATION

The findings of the study indicate that Red Onion (Allium cepa) peel extract can be used

as a potential organic component to commercialized fungicides or apply the extract itself

particularly to eliminate sugarcane fungal pathogens for the advantage of agricultural industries.

Moreover, the extract was found to be more effective compared commercial fungicide.

17
RECOMMENDATION
To further contribute novel ideas regarding different problems in society, research can

support better solutions to the gaps in the current state of the field. This is for the advantage not

only for the civilization of today but for future generation as well. Still, this research study seeks

to be modified to address different problems associated with the field of agriculture. The

researchers wanted to recommend the following:

1. Focus on characterization, purification and isolation of specific compound in the red

onion peel.

2. Directly apply the concentrations to the sugarcane making an in-situ study.

3. Determine what other fungal pathogens the Red Onion (Allium cepa) peel extract can

inhibit.

4. Determine what other extracts can hinder the growth of Sporisorium scitamineum.

18
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Qo

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APPENDICES

Raw Data
Radial Growth Per Day During Five Day Incubation
TEST GROUP DAY 1 DAY 2 DAY 3 DAY 4 DAY 5

Rep1 Rep2 Rep3 Rep1 Rep2 Rep3 Rep1 Rep2 Rep3 Rep1 Rep2 Rep3 Rep1 Rep2

S. scitamineum Treated with Fungicide NVG NVG NVG 8.56 9.06 8.20 9.35 9.45 9.12 13.76 13.50 13.76 18.17 18.01

S. scitamineum Treated with 30% OPE

NVG NVG NVG NVG NVG NVG 7.03 7.10 7.05 9.44 9.80 10.75 10.84 12.53

S. scitamineum Treated with 20% OPE

NVG NVG NVG 7.23 7.59 7.50 9.12 9.68 9.13 11.82 10.08 9.62 12.33 11.44

S. scitamineum Treated with 10% OPE

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 S. scitamineum Treated with No Treatment 7.49 7.63 7.05 8.26 8.60 8.52 18.57 18.64 18.41 29.38 34.23 30.88 39.82 39.81

26.75 26.48 25.95 34.30 31.15 32.56 45.12 45.80 46.16 65.15 68.18 64.34 77.75 77.65

Note: NVG- No Visible Growth

& OPE- Onion Peel Extract

Raw Data
Average Radial Growth Per Day During Five Day Incubation
TEST GROUP DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 Average
Mean Score
S. scitamineum Treated NVG NVG 7.06 10.00 10.96 5.60
with 30% OPE
S. scitamineum Treated NVG 7.44 9.31 10.51 14.60 8.37
with 20% OPE
S. scitamineum Treated 7.39 8.46 18.54 31.50 38.14 20.81
with 10% OPE
S. scitamineum Treated 26.39 32.67 45.69 65.89 78.60 49.85
with No Treatment
S. scitamineum Treated NMG 8.61 9.31 13.67 18.31 9.98
with Fungicide

Table 1
Phytochemical Screening of Red Onion Peel Extract
QUALITATIVE PHYTOCHEMICAL FOR CONSTITUENTS

Sample Flavonoids Saponins Amino Acids Proteins Tannin

Red Onion
Peel
Ethanolic + + + - +
Extract
Note:
Result are specifically related to samples as received.
(-) may denote compound to be present in trace amount.

Table 2
Composition of Commercialized Fungicide Benomyl 50WP

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 Active Ingredient:
Benomyl 500g/kg
Inert Ingredients 500g/kg
Table 3
Radial Growth after the Introduction of Red Onion Peel Extract
Radial Growth (mm) Averag
N e
Std.
Treatment (per Radial
Day 1 Day 2 Day 3 Day 4 Day 5 Deviation
pay) growth
per day
30%
3 NVG NVG 7.06 10.00 10.96 2.19 .304
concentration
20%
3 NVG 7.44 9.31 10.51 14.60 2.92 .945
concentration
10%
3 7.39 8.46 18.54 31.50 38.14 7.63 .580
concentration
50mg
Commercial
3 NVG 8.61 9.31 13.67 18.31 3.66 .078
fungicide (CAT
Benomyl 50WP)
No Treatment 3 26.39 32.67 45.69 65.89 78.60 15.72 .311

Note: No Visible Growth (NVG)

Table 4.a
Significant Difference Among Treatments
Sum of Df Mean F Sig
Squares Square
Between Groups 9410.433 4 2352.608 329.718 .000
Within Groups 71.352 10 7.135
Total 9471.786 14

Table 4.b
Post Hoc Analysis in Scheffe
Dependent Variable: Radial Growth in Day 5

Linear Comparison Mean Difference Sig.

30% Extract - 20% Extract -3.65 .610

30% Extract - + Control -7.53 .073
30% Extract - 10% Extract -27.18* .000
30% Extract - - Control -67.64* .000

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 20% Extract - + Control -3.89 .555
20% Extract - 10% Extract -23.54* .000
20% Extract - - Control -63.99* .000
+ Control - 10% Extract -19.65* .000
+ Control - - Control -60.11* .000
10% Extract - - Control -40.46* .000
Note: * The mean difference is significant at 5%.
Table 5
Correlation between the concentration of red onion peel extract and radial growth
Concentration of
red onion peel N Mean Std. Deviation
extract
30% Extract 3 10.96 1.52
20% Extract 3 14.60 4.73
10% Extract 3 38.14 2.90
Correlation N P
-1.000* 9 .000

25