The Bigger the Bean

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Dec 6, 2023

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1 Effects of Bean Size on Oviposition: Bean Beetles Payton Weinstock Florida State University BSC2011L: Biological Science II Lab Professor Dani Davis April 17, 2023

2 Abstract Bug molecular biology has long been focused on genes that are involved to ovulation. Oviposition is the term used to describe the act of depositing eggs. The process of oviposition, which involves egg laying and egg hatching, is how most insects reproduce. It is crucial to consider several environmental characteristics, such as indicators given off by the beans, such as aroma, texture, and temperature, while choosing the best location for oviposition. The female will oviposit where she thinks her offspring will perform best after carefully weighing a variety of criteria. Since we'll be utilizing the same two beans in two different sizes, we'll concentrate primarily on bean size. Originally from Africa and Asia, bean beetles, or Callosobruchus maculatus, are agricultural invasive pests that now reside in temperate and subtropical climates worldwide. Because they are rich in fiber, protein, B vitamins, magnesium, and iron, lima beans are a fantastic source of nutrition. Adults spend their brief lives mating and laying their eggs on legumes because they don't need food or water. We started our experiment by selecting a sample of bean beetles that had previously been put in a plate with the bottom lined with black eyed peas, which resemble lima beans in color. In order to begin assembling our setup, we gathered three circular petri dishes and filled the bottom of each with 14 little and 14 large lima beans. Three females and 2 males were present in each dish. The relationship between the size of the lima bean and the quantity of eggs laid was investigated using a chi-square test. Whether bean beetles chose to deposit their eggs on small or large beans had no difference. The chi-squared test findings show that this is not statistically significant between lima bean size and the probability of oviposition. As a result, the null hypothesis cannot be rejected. Introduction

3 Oviposition-related genes have long been a center of bug molecular biology. The process of depositing ova is referred to as oviposition. Most insects procreate through oviposition, which involves egg laying and egg hatching. When a female discovers an appropriate location to oviposit on its host, she deposits several eggs. During the course of this procedure, numerous genes regulate the female's actions both behavioral or physiological, such as ovulation, keeping temperature on the egg surface, and preserving a steady internal climate in the eggs (Hu et al., 2019). Beneficial insect longevity and health are dependent on successful oviposition and egg hatching (Reddick et al., 2018). When determining oviposition choice, it is important to consider different environmental factors such as cues emitted by the beans, such as scent, texture, or temperature (Delobel and Sembene, 2000). Chemical substances on the bean surface (fatty acids and alcanes) play an essential part in oviposition behavior production. Because these chemicals are linked to tegument pigment and toughness, they may explain the substantial variation in oviposition decision (Parr et al., 1998). After careful consideration of many factors, the female will oviposit where they believe their offspring will have the greatest performance. We will be focusing specifically on bean size as we will be using the same two beans of two different sizes. This will make it possible to control for the variable as both beans are the same in color and nutritional contents. Bean beetles, Callosobruchus maculatus , are agricultural invasive insects native to Africa and Asia that now inhabit the temperate and subtropical regions of the globe. The southern cowpea weevil is another name for this species. This species' larvae eat and mature solely on the seed of legumes, hence the term bean beetle. Lima beans are a great source of nutrients as they contain fiber, protein, B vitamins, magnesium, and iron. Adults do not need sustenance or water and spend their brief existence (one to two weeks) mating and depositing eggs on legumes

4 (Kergoat et al., 2007). Because of their short reproductive cycle/life cycle and their ability to not require food or water because they rely solely on getting nutrition from the bean, makes them a good model organism. This makes it possible to conduct an experiment within a short period of time and requires little to no maintenance. Since we are using two of the same beans (lima beans) of differing sizes, we will be testing whether Callosobruchus maculatus prefer to oviposit on a smaller or larger lima bean. Another study like ours involving Melitaea cinxia , a species of butterfly, was conducted in an effort to identify the relationship between plant size and oviposition. The results of the study found plant size to be an important stimulus, with M. cinxia favoring large plants for oviposition as opposed to smaller ones (Reulder et al.). As is relates, we expect to see more eggs laid on the larger bean as they are denser nutritionally and have a larger surface area, allowing the bean beetles to lay more eggs. Methods We began our experiment by choosing a sample of bean beetles that were previously placed in a dish with black eyed peas, which are similar in color to lima beans, lining the bottom. Our bean beetles had an expected hatch date of March 21 st : We collected them on March 27 th .The dish contained ten females and ten males and was initially kept in an incubator at 28.8 degrees Celsius. At first sight, all bean beetles were active and alive. Females appeared as a darker color with white spots and males appeared with a red/brown coat and no spots. To start putting together our set-up, we gathered three round petri dishes and covered the bottom of the dish with 14 small lima beans and 14 big lima beans. We moved the beans around on the dish so that they were equally dispersed. We then obtained an aspirator which is a common instrument to collect small insects. The aspirator consists of a metal straw to intake the

5 insect, a rubber straw in which we stuck on, and a filter that prevents beetles from being sucked up into the tube. There is also a vial attached that the bean beetles get collected in. One vial has a female symbol and the other has a male symbol. We then began collecting bean beetles using the aspirator where we collect three females in the female vial and two males in the male vial then put them in our petri dish. We conducted this same process for our two other treatments. After this was completed, we had a total of 15 bean beetles with three females and two males in each lima bean lined petri dish. The sides of the dish were taped enough to keep the bean beetles from escaping. They were then left for two weeks while females began their oviposition preference process. We will use a microscope to count the number of eggs using a microscope to determine beetle oviposition preference after the two weeks. We will count the number of eggs on both the large and small bean. Since we are collecting categorical data, we will use a chi-square test to make inferences about the results. Results A chi-square test was performed to examine the relation between lima bean size and the number of eggs laid. There was no significant relationship between whether bean beetles preferred to lay eggs on a small (M=3.33, SD= 0.577) or large lima bean (M=5.00, SD= 5.000) , X 2 (1, n=9) = 1.000, p=0.3173; Fig 1).

6 Discussion Our initial study was to investigate the probability of the female bean beetle's oviposition selection. This decision is most likely related to how successful the offspring will be. Based on the results of the chi-squared test, there is no statistically significant difference between lima bean size and likelihood of oviposition. Therefore, we fail to reject the null hypothesis. Another study was completed to answer our same hypothesis. Although this study used a different kind of bean, they were similar in size to the lima beans that we used. Bischoff et al. 2017 findings indicate that, when compared to the number of eggs deposited, smaller beans have a higher proportion of adult beetle progeny than larger beans, yet there is not enough data to make the results statistically significant. Our studies relate as they were both under the same conditions such as given two beans to choose to lay eggs on, incubated, and left for two weeks to oviposit. One difference is that our bean beetles initially came from black eyed peas whereas their substrate in the study were mung beans. This can attribute to why we had similar results and can be a limiting factor. Because of the fact that they came from beans similar in size to the small

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