How Different Levels of Capsaicin Effect Fungal Growth

Introduction
Capsaicin is a defensive compound found in chilis. The peppers vary in levels of capsaicin. However,
experiments and tests show that peppers with higher levels of capsaicin are spicier than peppers with
minimal to no capsaicin (Borrel, 2009). The capsaicin helps the plants to fight off natural predators such
as insects and fungus. The higher the capsaicin levels the less insects can get to the seeds of the plant
which diminishes dispersal (Schussler, 2014). After reading articles and learning more about capsaicin, it
led us to ask the question: How would fungus taken from plants with high capsaicin levels versus fungus
taken from plants with low capsaicin levels act when placed on a petri dish with capsaicin?

This question led us into our hypothesis that if fungi is grown on plates with capsaicin, then the fungus
retrieved from spicier peppers such as habaneros (H-fungus) will grow more than a fungus retrieved
from a tomato plant (T-fungus) because the fungus is better adapted to the environment with capsaicin.
Based off our hypothesis, we predicted that the higher the capsaicin levels, the more significantly
different the growth size will be between the H-fungus and T-fungus. Fungus isolated from habaneros
will grow more in environments with capsaicin then fungus from tomato plants. Based off our readings
from the articles, we can confirm that the hotter the pepper the more capsaicin is in the plant.
Therefore, if we collect a fungus from a high-level capsaicin plant, that tells us that the fungus has
evolved to combat capsaicin, so if we place the fungus that can combat capsaicin on a petri dish with
high levels of capsaicin, then the fungus will continue to grow. However, if we retrieve fungus from a
tomato plant with no capsaicin and then place it on dish with capsaicin, it will have no defense
mechanisms to combat the capsaicin. Therefore, it will not grow very much if at all. In this experiment
the null hypothesis would be that both fungi grow equally on the dishes with varying levels of capsaicin.

Methods
Experimental Design:
For our experimental design, the independent variables were 1. Fungal spores from habanero peppers
and 2. Fungal spores from tomatoes. The dependent variable was the fungal growth for each plant with
varying levels of capsaicin. We used eighteen total petri dishes with varying levels of capsaicin for the
experiment. There were three capsaicin groups: C0 with no capsaicin, C1 with one gram of capsaicin,
and C4 with four grams of capsaicin. For each capsaicin group, there were six dishes: three for H-fungus
and three for T-fungus. We placed fungal spores on each dish in a sterile environment. We then taped
the petri dishes closed and labeled the dishes with group. Name, class time, level of capsaicin, and
temperature incubated. Finally, we put them in a 28°C dark incubator.

Data Collection:
We removed T-fungus from the incubator after three days and H-fungus from the incubator after five
days. We removed T-fungus sooner due to how rapidly it grows. After retrieving the dishes, we
measured diameter of the fungal growth for each dish from the largest point of diameter that was
visible. We could then compare the diameters for each fungus along with the different levels of
capsaicin present. At this point, we could begin to test if our hypothesis was correct. We needed to find
any statistical difference in the data to support or reject our hypothesis. A statistical difference would be
a p-value<0.05. We got this p-value from t-test.
Results
Question: How does the growth rate of different species of fungi isolated from plants with capsaicin
and plants without capsaicin differ when placed in an environment with capsaicin

Figure 1

Figure 1 shows the difference between H-fungus and T-fungus when placed in dishes with varying levels of
capsaicin. The number above each bar shows the exact diameter of the fungus.

In Figure 1 the x-axis represents the amount of capsaicin in each of the three petri dish groups. The y-
axis represents the diameter(mm) of the fungal growth in each petri dish after the allotted time. The
figure shows that the fungal growth between habaneros and tomatoes was relatively similar. We found
the p-value for each amount of capsaicin present. The values found were: C0:0.60643. C1: 0.051429, and
C4:0.615793.

Discussion

Hypothesis:
Based on the results from our experiment, we were able to conclude that our hypothesis was rejected.
Our hypothesis stated that fungi retrieved from plants with high capsaicin levels would be better suited
to survive in an environment with capsaicin. However, our data showed us there was little difference in
fungal growth between H-fungus and T-fungus. There was no significant difference in the diameters
between the fungi. This can be supported by the p-values being greater than 0.05. This means that
fungus retrieved from any sort of plant could grow when placed with capsaicin. The fungus can evolve to
be better suited to grow with capsaicin.

Analysis of Data:
The data that was gathered from the experiment is accurate from what we can conclude. However,
there is always room for human error. For example, it is a possibility that the diameter measurements
could have been off for the fungal growth. In addition, we did have one error in the experiment. One of
the petri dishes was taped upside down, so the fungus grew on the lid of the dish. Although this sample
was a little bigger, we are unsure of the impact it may have caused in the data. There were no outliers in
our data set that impacted the results. However, there is always room for improvement. If my group
were to do this experiment again, we would make sure all the dishes are upright, and we would also try
to take more accurate measurements of the fungi.

Implications:
Our results do make sense when it comes to background information and previously known information.
Capsaicin is a chemical compound that naturally fights off fungi. However, fungi have learned to evolve
to combat that in what’s called the evolutionary arms race. Our data shows that the amount of capsaicin
and the origin of the fungi had little to do with the growth of it. Therefore, the bigger picture shows that
capsaicin would not be agriculturally beneficial to farmers. IT would not effectively prevent fungal
growth. This means that farmers can stop putting non-helpful chemicals into or foods to prevent fungal
growth. The fungus will find ways around it through evolution.

Further Investigation:
In the future, I would like to test fungal growth with varying capsaicin levels on much longer periods of
time. I would like to see how the fungus from a plant with capsaicin is affected long-term versus how a
fungus from a plant with no capsaicin is affected long-term. If the fungal growth is significantly different,
is it because of the type of plant the fungus came from, is it due to the environment, and what other
factors could affect it?

Works Cited

1. Borrell, B. 2009. What is So Hot About Chili Peppers? Smithsonian. 80

2. Schussler E. 2014. Biology 159 Lab Manual: Skills of Biological Investigation (General
Biology,University of Tennessee - Knoxville). bluedoor, LLC: Minneapolis.  
3. SITNFlash. “The Complicated Evolutionary History of Spicy Chili Peppers.” Science in the News,
12 Aug. 2013, https://sitn.hms.harvard.edu/flash/2012/issue131b/#:~:text=Capsaicin
%20probably%20deters%20insects%20much,of%20plants%20that%20made%20spice.