Académique Documents
Professionnel Documents
Culture Documents
The main goal of this study was to utilize the antifungal activity of corn (Zea mays) cob to inhibit
the growth of Fusarium oxysporum, a fungi known to be resistant to most available drugs. The
researchers obtained the extract using the maceration method to release the necessary
phytochemicals for inhibition. Three different concentrations for the extract were employed for
antifungal testing which are: twenty five percent, fifty percent and seventy five percent. There
were three trials for each concentration in correspondence to the Randomized Complete Block
Design. The zones of inhibition for all concentrations were equivalent to zero. Using the
Analysis of Variance and Pearson-R Correlation, at the conventional level of significance, the
computed f and r-values are smaller than their respective tabular values. Based on the
abovementioned result, the decision to accept all null hypotheses were formulated. It was
concluded that the various concentrations of the corn cob extract have no significant effect on the
size of inhibition. Furthermore, there is no significant relationship between the extract and the
fungi. Numerous researches have reported the presence of phytochemicals in corn cobs, however,
this study proved that it is not sufficient to inhibit the growth of Fusarium oxysporum. In
accordance to the results of this study, the researchers highly recommend finding other plant
waste materials with potential antifungal properties, applying the extract to other fungi and
discovering other methods to stimulate the antifungal activity of corn cobs. It also proposes the
utilization of the extract for other areas of concern such as antibacterial and anticoagulation
fields.
Antifungal Activity of Corn (Zea mays) Cob Extract against F. oxysporum (Fusarium
oxysporum)
The Philippines is an agricultural country with a land area of 30 million hectares, 47
percent of which is used for farming. Contrary to the massive production of crops, several
problems are besetting the farming industry such as the spread of pathogenic microbes. Crop
diseases have an effect on the income of farmers and the health of consumers. Several
researches have been conducted in an attempt to provide low-cost and efficient solutions to crop
Corn is the second most important crop in the Philippines that contributes significantly to
the country’s economic development. Corn wastes are considered one of the top agricultural
wastes in the Philippines. The combustion of these wastes pose serious risks on the environment
and the health of farmers. The cob is the part of the corn ear on which the kernels grow. This
part will become the waste product once the kernels have been consumed. Phytochemical
studies revealed that corn cob contains liquid crystalline oils, flavonoids, phenolic compounds
and this anthocyanins which result in the anti-inflammatory, antibacterial and antioxidant
properties of the cob. Carvacrol, thymol and menthol are phenols used chiefly as fungicides,
moisturizers and disinfectants to treat fungal infections such as eczema and rashes
(botanicalonline.com).
According to Dong et. al (2014), three of the most commonly known by- products of corn
(Zea mays) are used in different herbal medicine due to its rich chemical deposits within it and
was proven to be safe. Various studies show that the three are rich in phenolic compounds,
flavonoids, kerotinoids and different antioxidants and other antibacterial properties. Stigma
maydis is proven to have the richest contents among these parts. It contains exceptional
bioactivities which are due to hydroxyl groups found in these phenolic compounds. Corncobs
have almost the same properties with the former but the study also showed that anthocyanins
were present which is a type of flavonoid. On the other hand, corn cobs and corn husks are easier
to obtain than stigma maydis because of a higher yield in production and most of these are
thrown out.
Maize grain is the third most consumed cereal, surpassed only by rice and wheat. It is
ranked as one of the cereals with the highest content of phenolic compounds. In maize grain, the
phenolic acids: ferulic acid and p‐coumaric acid predominate, which are highly abundant in their
bound forms. However, other phenolics such as anthocyanins, flavonoids, and flavanols have
been identified in colored maize grains. Flavonoids possess important biological activities such
Fusarium oxysporum
forms, each specific to a unique plant host. F. oxysporum causes fusarium wilt, a vascular
wilting disease characterized by root rot, seed blight, rotten fruits and discolored leaves followed
by the impairment of the whole plant (Campbell et al, 2014). According to Gutierrez (2016),
approximately 15,000 hectares of land have been affected by the Fusarium wilt that is considered
as one of the first disease that affects perennial and annual plants to have spread globally. Dry
weather and low moisture accelerates the development of F. oxysporum. Attempts of complete
eradication have only produced insignificant results since mycelia growth displayed high
adaptation ability in different environmental conditions. The most effective solution is soil
sterilization, a method that requires a lot of equipment and is of long duration. Alternatively,
some farmers use commercial fungicides with limited effects on the fungus and negative effects
According to Puig (2014), Fusarium wilt is a global threat to all banana growing areas
around the world. The massive losses of bananas, specifically ‘Cavendish’, in Davao region’s
banana industry is caused by this disease. Because of this, a huge area of lands in Davao del
Norte and ComVal were abandoned. One of the causal organisms is Fusarium oxysporum f. sp.
cubense Tropical Race 4 (Foc TR4). This affects all banana cultivars growing in the region.
Fusarium wilt in the region remains to be a problem in some Asian countries, Australia, South
Fusarium wilt in Cavendish bananas around Davao on the southern Philippine island of
Mindanao was caused by Foc VCG 01213 that was found both on the Mindanao and Luzon
islands, and was isolated mainly from the Silk cultivar Latundan. VCG 0122 was related with the
Cavendish cultivar Grande Naine in the Davao del Norte area. Three different VCGs were
isolated from Grande Naine, namely VCGs 0122, 01213 and 01216. (Mostart et al, 2017)
Numerous crops have been known to be host plants for Fusarium species causing soil-
borne diseases. Though extensive studies have been performed, little is known about the fungus
stimulating or inhibiting properties. In the present study, the effect of flavonoid compounds on
the germination and fungal growth of soil-borne pathogen Fusarium oxysporum was observed.
oxysporum, whereas the other flavonoids were inactive at different concentrations. Individual
flavonoid concentrations resulted in a small increase of fungal growth. However, the lowest
flavonoid concentrations showed an inhibiting effect on fungal growth. This suggests that low
flavonoid concentrations exhibit antimicrobial properties against F. oxysporum (Steinkellner &
Mammerler, 2016).
The Philippines has and continues to undergo an economic crisis when it comes to the
agricultural sector. The growth and quality of the crops can be affected by a lot of factors. One
of these is the susceptibility of the plant to different pathogenic microbes. Fusarium oxysporum
is a fungus that enters the host through the roots and grows on the xylem, blocking the vascular
and preventing the transport of water and nutrients. Species in the Solenaceae family such as
bananas, eggplants, pepper, potatoes and watermelons are the most susceptible to fusarium wilt,
commonly known as Panama disease. This fungus can be dormant for 30 years before resuming
virulence and is resistant to most antimycotic medications due to its rapid mycelia growth and
ability to survive in the absence of effective biological, chemical and physical control measures
(Ploetz, 2015).
well as national economy. The resistance of Fusarium oxysporum to a wide variety of chemicals
has stimulated the search of new alternatives for control measures. The use of plant extracts are
one of the most promising, effective, safer and eco-friendly method to eradicate the pathogen
from ripe tomato. Out of 9 extracts, it was found that almost all extracts at 25% concentration
were effective in reducing the mycelium growth of Fusarium oxysporum f. sp. lycopersici. The
highest inhibition (87%) of mycelium growth of this pathogen was observed when treated after
the plant extract of Calotropis proceraon at 25% concentration and lowest inhibition (27%) of
this pathogen was observed when treated after the plant extract of Momordi cacharantia on at
emerging opportunistic pathogen of human and other animals. Antifungal susceptibility test
revealed the concentration dependent growth inhibition of lawsone against F. oxysporum with
minimum inhibitory concentration (MIC) at 100 μg/mL. Ultra-structural analysis indicates the
prominent damage on cell wall of the mycelium after lawsone treatment, and imples that it could
increase the membrane permeability and disintegration of cells leading to cellular death.
Lawsone can induce the reactive oxygen species (ROS) level in the filaments of F. Oxysporum
which activates the autophagy related pathways due to the oxidative stress. Results showed the
dependent manner. F. oxysporum infected zebrafish has recovered after lawsone therapy as a
topical treatment suggesting that lawsone is a potential natural antifusariosis agent (Dananjaya,
et al., 2017).
chemical constituents in organic materials. Phenolics are secondary metabolites that are found in
several fruits, grains and different parts of plants. It is considered to have a wide range of
protection. The solvent to be used can greatly affect the amount and type of phenolic compounds
to be extracted. Ethanol and methanol with different concentrations are commonly used
whenever phenolics, flavonoids, kerotenoids and tannins are extracted on plants. The study used
100ml of water, 50% ethanol, 80% ethanol, 50% methanol, 80% methanol and ethyl acetate to
test each in 2g- sample powders of three by- products of corn (corncobs, cornhusk and stigma
maydis) in for a day. It was then extracted via ultrasonic cleanerat 40°C thrice, and then
concentrated through a rotary evaporator. After different tests it was proven that 80% ethanol can
be used to effectively extract more from the by- products of corn (Dong et. al, 2014).
According to Saikia and Mahanta (2013), it was reported that the phenolic content and
flavonoid content of cooked vegetables are different from the uncooked form as it could have
effect like destruction, release, and transformation of the phytochemicals in the vegetable.
Application of heat could change the structural integrity and cellular matrix of vegetables when
cooked. This means it can have either positive and negative effects on the total flavonoid content
of the vegetable. In the study, it was shown that boiling the vegetable lead to a decrease in the
The goal of extraction is to separate soluble plant metabolites, leaving behind insoluble
cellular marc (residue). The initial crude extracts commonly contain a complex mixture of
of extraction used in medicinal plant research. It involves soaking powdered plant materials in a
stoppered container with a solvent at room temperature for a minimum period of 3 days. This
process intends to soften and break the plant’s cell wall in order to release soluble
phytochemicals. The mixture is then pressed or strained by filtration. Conventionally, the choice
of solvent will determine the type of compound extracted from the samples (Azwanida, 2015)
in-vitro Antimicrobial Activity from Fruit of Momordica charantia L.”, research has shown that
“longer maceration time often leads to chemical changes, particularly oxidation of phenolic
compounds such as flavonoids. It was the reason why there were no antifungal activity observed
for hexane that was macerated at 12 and 24 hours. But the presence of other phytochemicals that
conferred the activity might be the reason why mild antifungal activity was observed at 48 hours
Over the past few decades, antifungals have decreased in effectivity due to the emergence
of drug-resistant fungi and toxic reactions. F. oxysporum is resistant to most available antifungal
drugs. This study aims to utilize the antifungal activity of uncooked corn cobs to eliminate
Fusarium oxysporum. It will yield a significant difference between the sizes of inhibition and
concentration of the corn cob extracts. The researchers also predict that there will be a
significant relationship between the amount of corn cob extract and its antifungal activity against
F. oxysporum. The extract will be applied to a culture of the aforementioned fungi instead of the
infected plant to limit the effect of extraneous variables. This study provides a good chance of
reducing the cost of sterilization and the use of fungicides with little impact on the infected plant.
Problem Statement
1) There is a significant difference between the sizes of inhibition and the concentration of the
2) There is a significant relationship between the corn cob extract and the fungi, Fusarium
oxysporum.
The research study focus on extracting the uncooked corn cobs to test its antifungal
activity. It also focus on the potential of the extract against the fungi, Fusarium oxysporum. It
covers the use of laboratory and laboratory equipment such as florence flask and beaker.
Nevertheless, it does not seek to include the testing of the corn cob extract to an actual
plant affected by the said disease. It is also limited that the corn cob extract can completely
The study will benefit the farmers as the number of crops to be affected by Fusarium wilt
will be lessened. This will benefit them as a component to create a natural antifungal agent rather
than commercial antifungals. It will also benefit the community since the number of corn cob
wastes will decrease and be reused to agriculture in the locality. Future Researchers will also
benefit as they may come up with a more comprehensive study about the effect of the corn cob
This study will prove a safer and more effective alternative component to create
fungicides using corn cobs, lessening environmental wastes and producing an accessible natural
Method
Research Design
The appropriate experimental design for the study is the Randomize Complete Block
Design (RCBD). This design will be used to determine the cause of change by using successive
treatments. There will be randomization process to designate samples for the experimental group
of the study. RCBD implies that there will be repetition of the experiment and the treatments
used.
Measures
The study will use the Kirby-Bauer method, a common technique to measure the zone of
inhibition. This is the appropriate technique to measure the antifungal activity of corn cob extract
against F. oxysporum. The results will be measured by examining the area in which the fungi is
unable to grow because of the extract that will be applied. The areas will be measured in
millimeters and will undergo statistical treatment to see changes in significance of the different
concentrations.
Procedure
One thousand grams of corn cobs will be collected in the city of Santa Rosa in
Laguna. The corn cobs will be subjected to air-dry for an average of 3 days. After air-drying the
cobs were manually crushed using mortar and pestle. The researchers used manual crushing
instead of using millers to monitor the solubility of the cob and limit the contact of the sample
with contaminated materials. Approximately 1000 grams of pulverized corn cobs were then
soaked in 95% ethanol. The mixture followed a 1:3 ratio which implies that 250 grams of corn
cobs will be soaked in 750 ml of ethanol. This process was done under the following conditions:
at room temperature, using tightly-sealed containers at a duration of two (2) days. The initial
crude extract was then subjected to filtration using regent containers, funnels and filter papers.
Water bath was the final step for maceration which includes the exposure of the extract to
steam which allowed evaporation and condensation to take place. This was done to remove any
trace of ethanol in the extract. A stoichiometric amount of 25 ml of corn cob extract was placed
in an amber bottle and delivered to the laboratory in National Sciences Research Institute of the
University of the Philippines Diliman College in Quezon City for antifungal testing.
Microbial suspension was prepared from a 7-day old culture of F. oxysporum. 0.1%
microbial suspension by swabbing the agar surface. The cotton swab on an applicator stick was
dipped into the microbial suspension, rotated a few times, and pressed firmly on the inside wall
of the tube to remove excess inoculum. The swab was streaked over the whole agar surface. This
process was repeated two more times, rotating the plate 60° each time to ensure even
distribution. Three (3) equidistant wells were made on the agar plate using a cork borer. Two
hundred (200) μl of the sample in different concentrations was placed in each well.
The PDA plates were incubated at room temperature for 3 days. The clearing zones of
Results
Table 1 shows the summary table of significant difference between the sizes of inhibition
and different concentrations of corn cob extract at 25%, 50%, and 75%.
In 25% concentration, the sizes of inhibition of trials 1, 2, and 3 are all 0.0. In 50%
concentration, the sizes of inhibition of trials 1, 2, and 3 are all 0.0. In 75% concentration, the
The following data was calculated using Analysis of Variance. Based on the data shown
in the table below, the obtained F-value obtained was 0. The table showed the analysis of
variance of the gathered data at α=0.05 level of significance. The null hypothesis is accepted
This implied that there is no significant difference between the sizes of inhibition and
25% 0 0 0
50% 0 0 0
75% 0 0 0
Sources SS df MS
Between Groups 0 2 0
Within Groups 0 6 0
Total 0 8
F value Fcrit
0 5.1433
Table 2 shows the summary table of significant relationship between the corn cob extract
and the fungi, Fusarium oxysporum.The concentrations of the corn cob extract are 25%, 50%,
and 75% with three samples of each. In 25% concentration, the sizes of inhibition are all 0.0. In
50% concentration, the sizes of inhibition are all 0.0. In 75% concentration, the sizes of
in the table below, the obtained Rcomputed was 0. The table showed that the pearson-r correlation of
the gathered data at α=0.05 level of significance is equivalent to 0.632. The null hypothesis is
. This implied that the different concentrations of Corn cob extract and the sizes of
inhibition in each trial does not have any correlation with each other.
Table 2
Significant relationship between the corn cob extract and the fungi, Fusarium oxysporum.
1 0.25 0.0
2 0.25 0.0
3 0.25 0.0
4 0.50 0.0
5 0.50 0.0
6 0.50 0.0
7 0.75 0.0
8 0.75 0.0
9 0.75 0.0
H o
Discussion
This study utilized the antifungal activity of corn cobs to inhibit the growth of Fusarium
oxysporum. Antifungal tests have determined that the zones of inhibition for all concentrations of
the extract were equivalent to 0. All null hypotheses were accepted with respect to the statistical
treatments. This suggests that the corn cob extract has no significant effect on the growth of the
necessary to degrade lignocellulosic biomass to sugars. It also has a recognized ability to survive
without any pathogenic activity which implies that any attempt of inhibition might encounter
some difficulties. Contrary to previous studies regarding the pharmacological activity of the corn
plant, the phenolic acids and flavonoids present in corn cobs were not sufficient to inhibit the
growth of the fungi. According to a study by Mahmud, Nasrin and Podder (2018), the
studies, the researchers were able to conclude that three factors have contributed to hindrances in
the inhibition of the fungi: the method of extraction, amount of the extract and the tolerance of F.
oxysporum to ethanol.
The study only seeks to inhibit Fusarium oxysporum using corn cob extract. It does not
seek to include other parts of the corn plant for the extract. The researchers only used maceration
as a method of extraction in the said study. It focused on the potential of the corn cob extract
against Fusarium oxysporum that is tested on the laboratory not on the plant infected by
Fusarium wilt.
Implication of Findings to Theory and Practice
In this study, the inhibition of F. oxysporum using corn cob extract was not observed,
primarily due to the effect of extraneous variables and other factors concerning the methods and
samples. During the experimentation and analyzation processes, the researchers have identified
the significant contributions of the study. Specifically, it is the first research to have used corn
cob as an antifungal agent against Fusarium oxysporum. And it is also proposes the feasibility of
using the cob extract as an alternative, cheap component for natural fungicides directed to other
Similarly, the results of this study indicate that the fungi is widely resistant to almost, if
not all, drugs or organic materials with pharmacological properties. Studying other plant waste
materials, finding other ways to stimulate the antifungal activity of corn cob such as using
References
Plants,04(03).
doi:10.4172/2167-0412.1000196
doi: 10.4103/2231-4040.82956
https://pubs.ext.vt.edu/content/dam/pubs_ext_vt_edu/2811/2811-1020/2811-
1020_pdf.pdf
doi:10.1111/j.1470-9465.2004.00845.x
doi:10.5935/0103-5053.20140177
https://microbewiki.kenyon.edu/index
php/Fusarium_oxysporum
0.1371/journal.pone.0181630.
www.extento.hawaii.edu/kbase/crop/type/f_oxys.htm
Jain, P.K., et al. (2011). Current Status of Fusarium
Antagonistic sp. and Chemical Elicitors In Vitro. Fungal Genom Biology. Fungal
Torre (March 15th 2017). Phenolic Compounds in Maize Grains and Its Nixtamalized
of the banana Fusarium wilt fungus, Fusarium oxysporum f. sp. cubense, in Asia. PLOS
boiling and microwave cooking on the total phenolics, flavonoids and antioxidant
https://www.philstar.com/business/agriculture/2016/03/19/1564562/deadly-disease-puts-
philippines-banana-industry-under-threat
Curriculum Vitae