Biology Lab 020

BIOL1081L
Teresa Rust, Stephanie Vandergriff, Aaron Martinez, and Megan Walz

Oviposition in Toxic and Nutrient Rich Beans of Callosobruchus maculatus

Teresa Rust, Stephanie Vandergriff, Aaron Martinez, and Megan Walz
ABSTRACT
The purpose of this experiment is to identify whether female Callosobruchus maculatus,
bean beetles, also known as cowpea seed beetles can distinguish between poisonous and nutrientrich beans to oviposit, lay their eggs, on. The study used bean beetles from Asia who were raised
in mung beans and then introduced to a mixture of black-eyed peas and navy beans to study
whether they can distinguish between the two beans in order to have successful reproduction.
Black-eyed peas are nutrient rich, able to support the eggs laid on them, leading to successful
reproduction. They have a dark spot on them. The navy beans are similar in size and look very
similar in appearance except that they do not have a dark spot, but are toxic to the bean beetles,
not having the capacity to support the eggs, leading to unsuccessful reproduction. During this
experiment three beetles, one male and two female, were each placed into twelve petri dishes.
Ten of these dishes were replicates and two were control groups. In each of the replicate dishes
20 navy beans and 20 black-eyed peas were placed in large petri dishes containing the three
beetles. The results showed that there was not a significant difference in the amount of eggs laid
on the black-eyed peas versus the poisonous navy beans. These results support the idea that
female bean beetles cannot identify the nutrient-rich beans that will help their young reproduce

most successfully. The findings might be useful to farmers and those who run into bean beetle
pest control problems.
INTRODUCTION
Bean beetles (Callosobruchus maculatus) are small beetles that lay their eggs in beans.
These beetles are considered pests in agricultural regions because they oviposit their eggs on the
growing beans, therefore destroying the crop. The beetles only have one meal when they are
alive, and the bean they are laid on is that only meal. Consequently females have the important
job of choosing a bean that is both large and nutrient-rich. The bean beetles are usually laid on
mung beans or black eyed peas, other beans could be toxic to the beetles. Bean beetles have been
used in the lab recently for research because of their small size, short life cycle, and ease of
reproduction success. Recent studies have shown that there are three types of beans that these
bean beetles have shown the most success in reproducing on: mung, black eyed peas, and adzuki
beans (Bisby 1994). These three beans have been documented as being more successful with the
bean beetles than other various beans. In an attempt to see if the bean beetles detect a difference
between various types of beans, the purpose of this experiment was to see if bean beetles could
determine and choose a bean that was successful in supporting life over a toxic bean unable to
support life for the bean beetle. The null hypothesis was that there is no significant difference in
the number of eggs laid on the nutrient rich black-eyed pea verses the toxic navy bean. The
alternative hypothesis was that the bean beetles would lay more eggs on the nutrient rich beans,
the black eyed peas, versus a toxic bean, the navy beans.

METHODS
This experiment involving bean beetles was conducted at the University of Cincinnati led
by Dr. Jill Beyette and her Biology 1081 L students. The beetles used in this operation were
native of Asia and reproduced mainly on native mung beans. The experiment occurred over a
weeks time. Three beetles, two female and one male, were each placed into twelve petri dishes.
Ten of these dishes were replicates and two were control groups. In each of the replicate dishes
20 navy beans and 20 black-eyed peas were placed in large petri dishes containing the three
beetles. The 40 beans were all mixed together so that the decision of the female between what
beans she should lay eggs on would be purely a decision and not by chance such as if the two
different beans were placed on either side of the dishes. In one of the control dishes 20 navy
beans were placed in small petri dishes containing the three beetles, and in the other control dish
20 black-eyed peas were placed the same, also with the two female and one male bean beetles.
All of the beans used in this experiment were compared visually and each bean matched for its
size to try and keep all beans around the same dimensions in attempt to lower the amount of
variables affecting the outcome. After a week the amount of eggs laid in each dish, on blackeyed peas or navy beans, was counted and the results were gathered.

RESULTS
There was not a significant difference in the number of eggs laid on the black-eyed peas
verses the navy beans. The ratio between the number of black-eyed peas and the navy beans was
92:116. The average deviation was 2.4 (df = 9, p 0.5). The variance is 40.71 and the standard
error was 2.017. In analyzing the data t = 1.189. The two tailed t-table for p 0.5 value was
2.262 (df = 9). We accept the null hypothesis because 2.262 1.189. This means that there was

no significant difference between the number of eggs laid on the navy beans compared to the
black-eyed peas. Our experiment has a less than five percent chance of having these differences
caused by chance alone. Based on our data, bean beetles do not prefer to lay their eggs on blackeyed peas over navy beans.

Graph:
14
12
10
8
6
4
2
0
Navy

Navy

BEP
0
2
5
15
28
54
6
5
0
1

Navy
BEP

BEP

2
8
8
5
20
40
3
4
2
0

Mean
11.6
9.2

Standard
Error
5.741
4.067
4

t-Test: Paired Two Sample for Means

Mean
Variance
Observations
Pearson Correlation
Hypothesized Mean
Difference
df
t Stat
P(T<=t) one-tail
t Critical one-tail
P(T<=t) two-tail
t Critical two-tail

Navy
11.6
296.7111111
10
0.96321884

BEP
9.2
148.8444
10

0
9
1.189473481
0.132343145
1.833112933
0.264686289
2.262157163

DISCUSSION
The results from this experiment suggest that the bean beetles cannot distinguish between
nutrient rich and toxic beans. Our results were contrary to other researchers findings such as
Janzens whom stated, some bean species are very clearly toxic to Callosobruchus maculatus
larvae (1977). This may still hold true, but our results supported the stance that navy beans are
not one of the beans that is very clearly toxic to the bean beetles (Janzen 1877).
Other researchers such as Mitchell have focused more on the size of the bean that the
female chooses to oviposit on (1975). We tried to control the size of our beans to the best of our
ability by visually trying to match each bean so that they were all approximately the same size.
The dark coloring on the navy bean may have reminded the bean beetles of the mung beans
they were raised in because of the dark coloring spot. Mung beans are a medium to darker green.
This coloring may have played a role in how the bean beetles had no significant difference
between the navy beans and black-eyed peas.

In our experiment the bean beetles were raised on mung beans and put in petri dishes to
reproduce on black-eyed peas or navy beans, both unfamiliar to the bean beetles. Messina has
done research on whether bean beetles prefer to reproduce on the familiar mung bean or a new
bean (2004). While more eggs were laid onto the bean the females were familiar with having
come from the same themselves, they did recognize that the other bean could support their
offspring and also oviposited on the other bean (Messina 2004).
While our results supported the null hypothesis that the bean beetles did not significantly
favor either the black-eyed pea or the navy bean, Messinas research leads us to wonder if we did
more trials if the result would be different due to his results from his extensive research. We
would be interested in seeing this experiment on a much larger scale to see if the results would
change or stay the same.
The movement of the bean beetles, invading other ecosystems as invasive speeches poses a
problem not only for farmers, but also consumers (Mexican 2014). Governments are attempting
to control the spread of the bean beetle because of the harm they do to crops (Mexican 2014).
Currently they are dusting and spraying the plants with poisonous chemicals, but they are
looking for a less laborious and less poisonous way to control these pests for farmers (Mexican
2014). The research we have done could contribute to the research behind finding a better way to
control the spread of the bean beetle. Plants of both black-eyed peas and navy beans could be
intermixed and then the bean beetles would be less likely to migrate to new areas where this plan
is enforced because they do not survive as well as they did where they originally came from.
The bean beetles may not be able to tell between the toxic and nutrient rich beans so that they
would want to stay where they knew their offspring were thriving.

REFERENCES
Beyette, Jill. The Nature of Biology: Laboratory Modules in Working, Thinking, and
Writing like a Scientist. Cincinnati, Ohio: Van-Griner, 2014. Print.
Bisby, F.A., J. Buckingham, and J.B. Harborne. 1994. Phytochemical Dictionary of the
Leguminosae. Chapman and Hall, London.
Janzen, D.H. 1977. How southern cowpea weevil larvae (Bruchida Callosobruchus
maculatus) die on non-host seeds. Ecology 58:921-927.
Messina F.J. 2004. How labile are the egg laying preferences of seed beetles. Ecological
Entomology. 29:318-326.
"The Mexican Bean Beetle." Science News. University of Cincinnati Libraries. Web. 11
Sept. 2014.
Mitchell R. 1975. The Evolution of Omniposition Tactics in the Bean Weevil,
(Callosobruchus maculatus). Ecology. 56: 696-702.