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Student Research Symposium Abstracts - Spring 2017


BIOL 152, Badri Ghimire,

Different Types of Manure Have a Direct Connection to Plant Growth 
Alec Lindsey 

When gardening, manure is an essential part of growing plants, but does the type of manure truly matter? This project took 5 manures which included horse manure, cow manure, rabbit manure, and chicken manure, and tested it on 3 different plants.  My hypothesis was that different types of manure did not play a role in plant growth.   After 4 weeks of growth, the rabbit manure was significantly larger than the other manure types. The hypothesis was not supported due to rabbit manure proving to be superior.

Stomatal Density Varies in Creek Facing Leaves and Non-Creek Facing Leaves 
Madeline Mattos, Julissa Rendon, Nick Blackwell 

We tested whether or not stomatal density in the leaves facing the creek, and the leaves not facing the creek, on the same plant, varied. We took two leaf samples from four different plants all within eight feet of the Chico Creek on campus. We took nail polish and applied it to the under-side of the leaves, let it dry, and took clear tape to peel off the polish. The nail polish withheld the imprint of the leaf, including stomata, we then stuck it to a slide and examined under a microscope. We used 400x magnification using an ocular micrometer, which limited our field of view to 0.12 mm2. We counted the stomata in two different locations on the same leaf for additional data. We conducted a two-tail t-Test assuming unequal variances. Our P value was 0.01, which is less than the alpha of 0.05, meaning we had to reject our null that there is no difference between creek facing leaves and non–creek facing leaves in plants. In turn we accepted our alternative hypothesis that there is a difference in stomatal density in creek facing leaves than non-creek facing leaves in plants. Our results could support a future experiment regarding the hypothesis that stomatal density is responsive to environmental conditions.

The effect of pH on plants and their growth 
Victoria Miranda, Kedar Rozi, Lenore Robertson, 

How do coffee grounds affect plant growth in radishes? We hypothesized that coffee would provide the plant with more nutrients than just soil, predicting that watering the plants with water that contained ground coffee would cause them to grow taller and more quickly. There were 10 radishes planted for each sample group that were grown over a period of about 2 weeks. The experimental group did as we predicted, growing more than the control, but died in the end, disproving our hypothesis. The coffee grounds made the soil too acidic for the radishes in the experimental group.

Role of Water Source on Plant Height 
Walker Brown, Monica Quinton, Simon Burdick,, 

The effects that different types of water have on Lima bean growth was tested.  This was tested by using tap, filter, flowing, and pond water collected in Chico, CA.  The question is whether the different components of each water source will affect Lima bean height.  All Lima beans were germinated and grown in the same general location with the different water sources.  Plant height was recorded in centimeters every two days.  This experiment statistically supports that tap water has the largest effect out of all the water sources tested on Lima bean growth, showing an average of 76% more growth from the plants treated with tap water when compared to the average height of all other plants from different water sources.  It is concluded that water source does affect Lima bean height.

Sugar concentration affects arthropod attraction in different environments 
Yesenia Morales, Jamilleth Santillan, Jettefren Romero, Cynthia Mendoza,,, 

Many speculate insects are attracted to sugary substances due to palatability, although arthropods consume sugar to have an efficient metabolism and convert it into energy as the Krebs Cycle explains. We hypothesized that insect attraction will be higher in the liquid with the highest sugar content and not exposed directly to the sun instead of other substances with lower sugar content.  To test this hypothesis we researched the variation in insect attraction between liquids with different sugar concentration exposed to direct sunlight and the same liquids where they are not exposed directly to the sun. We examined on four different substances, each containing a different sugar concentration and placed them in different environments (sun vs. shade). We ran the experiment for nine days while recording data every three days. We accept our initial hypothesis since more insects were found in the liquid with the highest sugar concentration and less exposure to the sun.

The Heredity of Spot Density on Ladybugs 
Kayla Jelen, Juan Caracheo, Eunice Gomes 

The topic researched was the heredity of spots on ladybugs. Our hypothesis was that ladybugs inherit their spot density from their parents. We tested this hypothesis by employing artificial selection. The store bought containers of ladybugs, Hippodamia convergens as indicated by the store, were separated into two trials each with a control group containing randomly selected ladybugs, a group with less containing ladybugs with eight spots or less, and a group with more  that included ladybugs with ten spots or more. Each group from each trial was kept in a separate habitat. Each trial included twenty ladybugs in each group, allowing for a total of forty ladybugs in each group. These groups were then fed, kept in standard conditions and left to mate in order to produce a second generation. The parent groups were then compared to their respective offspring groups via three separate, t-tests which provided whether the number of spots in parents and their respective offspring were similar. The control groups and groups with more spots were found to be significantly different with a p value of 1.697 and t values of -0.132 and 0.019. The groups with less spots were not found to be significantly different with a p value of 1.697 and a t value of 3.001. Because two of the three offspring groups are significantly different with a confidence of 0.1 it can not be concluded that the spots on a ladybugs are genetically inherited from their parents.

Intermediate Disturbance in Upper and Lower Bidwell Park 
Brice Vanness, Olivia Puppo, Samuel Lowthorp, 

The Intermediate Disturbance Hypothesis states that at "intermediate" levels of disturbance, species diversity should be highest, and that at low and high levels of disturbance, species diversity will go down. We wanted to test if human disturbance, mostly along hiking trails, would have a small scale effect on species diversity of plants in a given plot. To test this, we marked our measured plots along various areas of hiking trails, and split up each plot into four equal sections. We then counted the number of each species, and how many members of each species were present in the given plot. To analyze our data, we used the Shannon Diversity Index, a commonly used diversity index in conservation biology. Once our data was analyzed, we noticed that there did seem to be a distribution that is predicted by the intermediate disturbance hypothesis. Parks such as Upper and Lower Bidwell seek to preserve the natural surroundings as much as possible, yet our findings show that human foot traffic has an impact on the biodiversity of plants here.   

The Effectiveness of Essential Oils as a Snail Deferent on Spider Lilies (Lycoris radiata)  
Caylea Molyneux, Haley Lowe, Kelli Thorup,, 

Various studies have shown that essential oils help defer various bugs and insects away from plants which protects them from damage. We tested the hypothesis that peppermint oil and cedarwood oil would decrease the damage rate on Lycoris Radiata caused by snails. We predicted that cedarwood oil would be more effective than peppermint oil. Through a duration of seven days spread out between two weeks, we examined and treated each plant every other day and counted the number of snails present and the number of holes in leaves of the three different plants with peppermint oil, cedarwood oil, and the control plant with water. After calculating our data, we accepted the alternate hypothesis that there was a difference in the damage rates between the control plant, peppermint oil plant and cedarwood oil plant, because the p-value was less than 0.05.  We also rejected the alternate hypothesis that the oils had an effect on the amount of snails present on the plants because the value was above 0.05.  Essential oils had a more positive effect decreasing the damage on Lycoris Radiata.

The Effectiveness of Batesian Mimicry Based on the Frequency of Mimic and Model Species 
Emily Ragan, Farah Yousif, 

Batesian mimicry is a phenomenon whereby unprotected prey species gain protection from predators by mimicking toxic or otherwise protected species. Predators learn to avoid models based on prior experience. An experiment was conducted over the course of three days using pita bread as the prey. We hypothesized that predators will learn that certain prey colors are not palatable as others based on the frequency of the models and mimics. The experiment was divided into five separate groups; blue, unpalatable blue, green, unpalatable green, and yellow prey. We used vinegar to make the bad-tasting models. Prey were set out at 8 am on trays of 25 along with a map of the randomized array, and removed at 6 pm. After running a chi-squared test, our results were less than 5.991, based on an alpha value of 0.05, on the first and second day. We concluded that feeding behavior is due to chance. On the third day, our results were greater than the accepted (X2= 6.09) therefore, the feeding behavior was different on the different colored prey. The morality of the mimics as well as the models is significantly dependent on the frequency of the unpalatable prey.

Variation of stomatal density in California Redbuds due to environmental stress 
Michal Hanson, Victor Cisneros, Ana Castillo,, 

The question we are trying to resolve is how environmental stress, such as proximity to water, affects stomatal density in California Redbuds. We inquired as to if being closer to a body of water creates more stomata in the leaves of this species. We tested our hypothesis by first measuring the distances of six plants from the creek and then collecting five leaves that are routinely exposed to the sun from each plant. We then got samples of the stomata from each leaf by painting the underside of them with clear nail polish and examined the samples under microscopes to count the average number of stomata per mm² on each leaf. Based on the results of the ANOVA, which states that the p-value is 2.02E-30, which is less than .05, we are 95% confident that stomatal variation is not due to proximity of water. Our data showed no clear correlation between distance from the creek and stomata in California Redbuds.

Does Distance from Chico creek affect species richness? 
Orlando Colbert; Alana Hyman; Colby Morris; Jade Seasate 

Water is known to give life to various species of plants and foster dense areas of specimens within nature. This supports the hypothesis that the density of species changes when the distance from the water source changes. We predicted that the species richness of a one meter by one meter plot of land is higher the closer it is to a source of water and gradually decreases the farther you get from the source. To test this, multiple one meter by one meter plots of land were chosen at intervals of 0.3, 1.5, and 4.6 meters away from Chico Creek, and then the number of species in the individual plots were counted to find results.  The lowest T-test value is greater than 0.05; therefore, the data is not statistically significant, and our hypothesis is not supported.  The areas we tested might have been affected by recreation more than the help of the water source, or the rain evenly distributed water for all plants this season.

Grubs Use of Batesian Mimicry 
Paulo Baylon, Kross Corbett, Parker White,, 

In this project experiment, Batesian mimicry is tested by using the consumption of grubs to determine whether or not is effective. We tested the hypothesis that birds will learn to avoid the unpalatable species. We predicted that birds will indeed learn to avoid the less favorable one by color. Camouflage has always been an effective defense mechanism for many species over the years. Many animals and plants are known to mimic others which are not preferable for consumption. Birds are often fallible to choosing whether or not a species are most likely to be palatable or not. We set up trays of 30 grubs made from lard, 3 groups of 10, separated by yellow (all palatable), green (¾ palatable) and blue color (¼ palatable). The results showed that the more palatable species were eaten as hypothesized. Because our chi-square value of 3.88 is less than 5.991, we reject the null hypothesis and conclude that the difference in feeding measured was due to chance alone.

BIOL 152, Bethany Prince,

Stomatal density and its relationship to distance from a water source 
Aracelie Agraz, Vannesa Lopez, 

Stomata are small pores that open and close by the functioning of guard cells found on the underside of a leaf in plants. Stomata play an important role in gas exchange. Water from a plant gets evaporated and released from the stomatal opening and in exchange carbon dioxide gas from the atmosphere enters the stomata and the process of photosynthesis occurs. The target of this experiment was to test whether plants near a water source had a higher stomatal density than plants farther away from a water source. Leaves were collected from two separate trees of the species Quercus lobata, also known as Valley Oak. Thirty leaves were collected from a tree with its roots in the Big Chico Creek and thirty leaves were collected from a tree 57.00 meters (187.01 feet) from the creek. The statistical results revealed that the tree directly in the water had a lower stomatal density than the tree farther from the water (P<0.05). Therefore, our hypothesis was not supported, since the tree farther from the source of water had higher stomatal density. This may be contributed to by the fact that its leaves were larger, while the leaves of the tree directly in the water were curled and smaller due to over exposure to water.

The Effect on the Western Squirrel's Eating Habits due to the Presence of Vinegar  
Cinthya Ledezma-Baeza, Kasandra Rockwell, 

Everyone with a bird-feeder knows that along with bringing in birds, it can also bring in some unwanted friends as well. Squirrels are known for getting into bird-feeders and taking the food, Home remedies have been explored and people found that if you use vinegar in the food, then they stop coming around. We soaked three types of nuts, peanuts, hazelnuts, and almonds, in vinegar for 16 hours and then laid them out for the squirrels to eat. By doing this we wanted to see if the nuts that weren’t soaked in the vinegar, were protected by the ones that were soaked in vinegar, due to Batesian mimicry. After analyzing our data we had a Chi^2 of zero. We found that the weather had a significant impact on our results and, because our Chi^2 was zero, that means our data is insignificant and there was a high probability of error so we couldn't reject or accept the null hypothesis.

Differences in Stomatal Density in Relation to Proximity of CSUC Creek  
Sahar Sandoval, Stephanie Montoya, 

The stomatal density in leaves influence the rate of carbon fixation as well as water loss. The higher the stomatal density, the more CO₂ that can be taken up. The ability of plants to make a developmental change based on environmental cues is caused by phenotypic plasticity. It occurs during the lifespan of an individual but is not a short term change. It was hypothesized that the stomatal density will be greater in plants that have access to water than the ones with no access. A sample of about thirty leaves from the plants not near the creek and plants in close proximity of the creek were collected, including the dry weight of both soils. After collecting the samples from both plant areas, the amount of stomata in each leaf of the near and far plants were observed with the use of a microscope. A t-test: two sample assuming equal variance, was used to examine the sample sizes to test the difference and concluded that there is a significant difference in both plants when in proximity to a water source. The plants near the creek attained a higher amount of stomata than the far plants. Distance to a water source strongly influences the amount of stomatal density in the near and far plants.

Temperature's Influence On Stomatal Density 
Annabelle Taylor, Amy Whitcomb, 

All plants require stomata, which take in carbon dioxide, in order to survive. However, can stomatal density vary within the same type of plant based on the temperature of the environment it lives in? Our belief is that as temperature increases, the stomatal density of a plant will increase, as well, because it needs more carbon dioxide entering in order to photosynthesize. We explored this idea by collecting leaves from a Vinca major plant in two different locations that had a noticeable difference in temperature. We then measured the stomatal density of thirty leaves from each location to determine whether there was a significant difference in the number of stomata. From this discrete data, we performed a t-test and determined that the stomatal density was greater for the leaves found in Chico, which had the greater temperature. Meanwhile, the leaves found in Magalia, which had the lower temperature, had an average stomatal density that was significantly lower. Based on this data, it was determined that temperature influences the stomatal density of a plant, as well as its carbon dioxide intake.

Water Source Effect on Coriandrum sativum 
Cynthia Vue, Monica Salas, 

The hypothesis of the experiment was that the creek water would promote more growth because the creek water is more natural. The methods include measuring the plants with a one foot ruler and watering the plants everyday. The conclusion was that there was no difference in the water sources pertaining to health because the p-value was 0.9 which is greater than 0.05.

A Survey of disturbance effects on observed alpha diversity in Upper Bidwell Park 
Donald Willingham 

In a decade where the environment is being put second after most other government and business affairs, it is important to keep sections of our world to house and protect wildlife as best as possible. Some of our greatest assets in this goal are parks such as we have here in Chico. After some thought it occurred to me that much of the Upper Bidwell Park looks the same, with tall grasses and occasional wildflowers, interrupted by a trail here or there. I was curious how the trails affect the diversity seen in the park, as it could be a source of disturbance that creates gaps in the tall grasses where plants can grow, or it could also be too great a disruptive force, allowing only the most resilient of weeds to survive. After counting the alpha diversity within ten quarter-meter square sections per three different distances from park trails in three different locations in the park I found that the data did not point to any conclusions about the park as a whole. The data show that alpha diversity along trails is significantly higher within areas of tall grass, while it is significantly lower in areas dominated by much shorter plants. It seems that park trails do create level playing grounds in some sections of Bidwell Park, while in others it stifles the ability of some plants to grow.

Watercress grown in soil vs watercress grown in aquaponics. 
Jose Serrano, Francisco Preciado, 

The common way to grow crops is that an individual generally plant the seeds in soil, but how do those compare to plants grown in water without soil. Aquaponics is a form of agriculture that reduces land consumption and produces both crops and fish in a sustainable system. My colleague and I decided to test if plants do better in an aquaponics system compared to soil in a downsized scale. The plant we decided to conduct this experiment is watercress (Nasturtium officinale) and the fish we used were common goldfish (Carassius auratus). N.offcinale was separated into two groups of 10. One group of 10 was left in soil, received regular daylight, and regular tap water from the city of Chico. The plants were left out for 6hr every day for two weeks. The second group of 10 plants were planted in the aquaponic system and were watered continuously with a water pump using tap water from the city of Chico. These plants were given 6 hours of red and blue LEDs as their daily primary light source. These plants were kept indoors for 2 weeks to prevent outside light to alter results. Initially there was not a significant gap in growth rates when comparing both methods of cultivation. The third day of growth started showing differences in growth, where in the aquaponics had seedlings compared to the soil where none have sprouted. During the progression of this experiment the growth rates seemed varied among individual plants but the aquaponic system saw faster growth overall when comparing the soil grown watercress.

Growth effects of fertilizer on Solanum lycopersicum 
Julia Reilley, Micheal Klemm, Diana Ledford,, 

The plant under study in this experiment is the common tomato (Solanum lycopersicum). This particular species is a part of the Nightshade family. The topic under study is understanding the growth effects that different fertilizers have on Solanum lycopersicum. This experiment can better help us understand how introduction of nutrients can vary the growth rate in plants. We determined how chemical nutrients differ from organic nutrients and how they compare to no nutrients at all. Mathematical statistics were used to identify correlation between the chemical, organic and control groups. An a-nova test will be conducted along with graphs that show the growth rate of the plants over a period of one month. Statistical analysis and p-value will be presented on the poster.

BIOL 152, Rachel Schleiger,

Comparing the effect of homemade fertilizer and store bought fertilizer on plant growth 
Alana Taylor, Joseph Garcia, Carly Martin,, 

There are factors that can influence the growth and health of a plant. Plant growth limitations include water, sunlight, temperature and nutrients. The goal for this study was to focus on the different levels of nutrients provided in three fertilizer groups: the control group, store bought fertilizer group, and homemade fertilizer group. We conducted this experiment to see the effects of each group on the growth and health of the Zinnia Giant Violet Queen flower. Our scientific question was whether there was variation in plant growth when using a homemade fertilizer versus a store bought fertilizer. Our three groups were Kellogg’s All Natural Gardening Soil (control), Kellogg’s soil mixed with Organic Plus All Natural Fertilizer (store bought), and Kellogg’s soil mixed with a homemade fertilizer (homemade). Our hypothesis was that variation in plant growth would be due to the hand selected combination and quantity of nutrients and minerals that will be included in the homemade fertilizer. We used 30 small pots and divided them into groups of 10. One seed was planted into each pot; there were 10 seeds per group and 30 seeds total. The ANOVA indicated no significant differences between groups (p=0.40075). Based off of these results, follow up T-tests were conducted to indicate specific variability between groups. Of these the strongest difference was noted between homemade versus store bought (p=0.0982). With more trials and a greater sample size, we could potentially see more of a difference between homemade and store bought fertilizer.

Northern California bird's feed size preference 
Christopher Hale, Perla Saavedra, Tonya Boscovich,, 

Different birds have different preferences when it comes to their food due to their beak size. Another variable in the preference is the risk factor, if birds are willing to come to the ground in order to get their desired food. This experiment set out to test both of these concepts on Northern Californian birds specifically the Californian Scrub jay (Aphelocoma Californica) and the American Robin (Turdus Migratorias).  The method that was used was placing small seeds and large seeds in four different bowls. One bowl of 80 grams of each kind was suspended 10 ft high in a tree and the other was placed at ground level. Bowls were weighed and replenished every other day. At the end of the experiment two t-tests were conducted, one comparing small and large seeds at 10 feet and one comparing small seeds at 10 feet and small seeds at ground level. Results indicated there is a preference in seed size (p-value=.0000416) but the second test proved that there is not a preference between seeds at elevation and on the ground  (p-value= .215345). Small seeds were preferred to larger seeds as we predicted. There is now a better of understanding of the birds in the test, their feeding habits, and preferences.

Phenotypic Plasticity of Blue Oaks in relation to distance from water source 
Nora Pizzella, Jillian Olivar, Kaleigh O’Brien, Andrea Coria,,,  

The focus of this experiment was to determine if Blue Oaks displayed phenotypic plasticity of stomatal density based on their distance from a water source(Horseshoe lake). Trees were divided into three different groups based on distance from the water source : 0-5 meters, 10 meters, and 20 meters. Each group contained three trees and leaf samples were collected from each tree. Leaves were be prepared for viewing under the microscope and numerical data for stomatal densities was collected. Once analyzed, the means were compared and an ANOVA test and a T-test were conducted to determine the statistical significance in the experimental findings. Results indicated support for the hypothesis that trees closest to the water source have the highest stomatal densities. With increased stomatal density, there is potential for higher productivity through photosynthesis with substantial water access.

Western Grey Squirrel (Sciurus griseus) feeding habits based on dietary preference 
Abrianna Sanders,Devin Sirls,Melissa Valencia,Rana Yousaf,,, 

Humans tend to modify their preferences for food based on the foods features. The sense of taste is instinctive to human’s daily lives that they seldom think about it. Animals have taste preferences for what they are consuming, similar to humans. It is this idea that the study aimed to analysis. The purpose of this study was to see if Western Grey Squirrels have food preferences and if they prefer natural foods grown from their own environment over human foods. The hypothesis for this experiment is that squirrels do have food preferences based on the different varieties of food. This selection may be caused by variables such as smell, texture, and taste.  The feeds used were bird feed, nuts, french fries, and fruit, all have a variety of different features. The prediction is that the squirrels will consume more french fries than the other feeds available. Methods for conducting this experiment included placing 100 grams of each feed in a park that was highly inhabited by the squirrels. The food is then monitored for 4 hours and weighed once every hour. This is repeated for two weeks. The Anova test for the entire experiment the F value (2.15) was less than the F critical (3.24), so the group fails to reject the null hypothesis as well. The data results do support the hypothesis that grey squirrels do have a preference for different food variables over their traditional food.

Mimulus guttatus elevation favorability in terms of height and fitness 
Carl Conserve, Nousheen Mahmud, Alyssa Myers, Inderjit Sarai 

Mimulus guttatus, commonly known as the monkey flower is found along stream banks and western Northern America and throughout most of America, predominantly in the west region of the United States. They are of the family Phrymaceae. Monkey flowers are found clustered in the “splash zone” of the pacific west, as it favors aquatic environments. Our experiment conducted at Table Top Mountain in Oroville California has supported the findings. Both the fitness and height of the plant were favored in the lower elevation as they had more exposure to water as it is readily available in low-lying ponds and crevasses. Our T-test values were 0.003591 for fitness and 0.027505 for height, which supports our hypothesis.

Variation in Stomatal Density of Camellia japonica Due to Proximity a Water Source 
Alex Fluitt, Claire Attias, Tyler Firebaugh,, 

Stomata are small pores located on the underside of a leaf that open and close to allow for gas exchange. When open, stomata allow carbon dioxide to enter the leaf for carbon fixation as well as allow water and oxygen to escape. The higher the stomatal density of a plant, the more quickly carbon dioxide can be taken in, and the more water can be released. This experiment tested the phenotypic plasticity of the Camellia japonica to see if the stomatal density would change depending on proximity to a water source. It was hypothesized that leaves on trees closer in proximity to a water source will have a higher stomatal density than leaves on trees further from a water source. To test this 15 leaves were collected from a Camellia japonica tree in close proximity to water (approximately 1.52 meters) and 15 leaves from the same type of tree located further from the same water source (approximately 15.24 meters), then obtained stomatal impressions for all 30 leaves. Results indicated that there was a significant difference between the stomatal density of leaves in close proximity to water compared to those in far proximity to water (p=0.003). Plants closer to a water source have a higher rate of water exchange for photosynthesis, which increases carbon dioxide exchange, and in turn requires plants to have a higher stomatal density.

Stomatal Density Variance Between Sun and Shade Leaves in Native and Non-Native Trees   
Lauren Howey, Erik Mechelhoff, Nelson Slen, Juan Dominguez,,, 

Stomata are small pores on the leaves of vascular plants. They control the movements of gases, and allow carbon dioxide to enter the leaf for photosynthesis and the release of water and oxygen. Leaf samples were collected from lemon (citrus limon), orange (Citrus sinensis), california buckeye (Aesculus californica) and big leaf maple (Acer macrophyllum) trees in Chico, California. The native buckeye and maple trees were expected to have a higher stomatal density average than the non-native lemon and orange trees. The stomatal density for the leaves located in sunny conditions were anticipated to have a higher stomatal density than leaves located in shaded conditions. For each species, 3 sunned leaves and 3 shaded leaves were collected. To calculate the stomatal density, a compound microscope at 400x magnification was used to count the amount of stomata present using a field of view area about 0.12 mm^2. The average stomatal density in the fruit bearing trees had a higher overall stomatal density in sunned leaves of 318.3 while the native tree average was 198.75. Shaded leaves in non-native trees had a higher overall stomatal density (267.9 ) over native trees (211.25). A T-test was used to evaluate the data, and the results concluded that there is no significant difference in stomatal density based on non-native species and native species (p value .39157792). Only the big leaf maple had a higher stomatal density in leaves found in shaded areas than leaves found in sunned areas of the tree. Overall, we can conclude that stomatal density in angiosperms varies greatly and no correlation was found in having less stomata present to bear fruit.

Goldenrod gall predation relations  
Samuel strawn,  Rain garcia, Daisy lopez-ramirez, Matt fritze,,, 

In this study the questioned relationship is the correlation between the gall size and predation for the Goldenrod plant. The study predicted that different gall sizes would have different level of predation as well as different predators. by gathered data based on what type of predator preyed upon different gall sizes an answer could be determined. In order to determine this, a number of goldenrods were planted and allowed for insect exposure. After gall formation plants were sorted by type of predator and then compared gall size. In our study, the study found that there was no significant predatory discrimination (p = 0.37). All sizes of galls had nearly equal predation. The conclusion reached was that gall predators are indiscriminate, the size of the gall has little effect on their interest.        

The Effects of Natural Sunlight versus Artificial Light on Plant Growth 
Sonia Alvarado-Ray, Kirshna Labayna, Natalio Plascencia, Brett Cote,, 

A plant’s light source is the one of most significant factors behind the stimulation of plant growth. This study aimed to test whether the type of light the plants were exposed to had an impact on the amount of growth the plants experienced in a two-week time span. Data was collected using two 50 seed sample groups of Zinnias (Zinnia); one group was exposed to natural sunlight, the other to artificial light via incandescent light bulb. Each plant was watered daily, with height recorded every other day. The final sample included 10 plants from each group, the average height of those which received natural sunlight was 5.82 cm, with a variance of 3.86 cm, while the artificial light group had an average height 8.35 cm, with a variance of 3.16 cm. Results supported the hypothesis that plants grown under different light sources will have different average heights (t-value = 3.39, critical t-value = 2.26, p = 0.00793). These results are important because it shows that plants can be grown indoors regardless of the outdoor weather conditions, as long as artificial light can be obtained.

BIOL 152, Stephanie Foster,

Does human traffic effect the frequency of plant herbivore defense mechanisms?
Giuseppe Aprile, Christian Garcia, 

Our overall goal was to explore if human traffic effects the amount of plant defenses used to combat herbivory.  We wanted to determine if human traffic effects the amount of latex secreted by Euphorbia peplus. We gathered 20 samples from two locations on Chico State campus and 20 samples from two locations in lower Bidwell park and gathered latex secretion from both sets of specimens. From our collected data we determined that the average mean of secretion in plants collected on Chico State campus was about a .53 difference between the two locations with a T-critical value of 2.023. According to our data and statistics we gathered form our T-test we can accept our original hypothesis and reject the null and thus, conclude that frequent human traffic has a significant effect on plant defense mechanisms and specifically latex secretion in Euphorbia peplus.

Effect of pH on Plant Growth 
Garrett Isley, Cole Gleaton, Aidan Ruiz,, 

We designed an experiment to determine the effect of pH on plant growth. We hypothesized that plants grown in a basic environment grow faster and taller than those grown in a neutral environment because acidic solutions denature plant enzymes and hinder growth. We took a sample size of twenty radish seeds and potted them in two separate trays, ten seeds watered with a basic solution and ten seeds watered with tap water, our control solution. We watered them once a day and observed the growth of the plants over a four week period. We then took the height of each plant in millimeters and determined our results. Our data showed that plants grown in a basic environment had an average height 37% higher than that of those grown in a neutral environment however, our statistical test results revealed that there is not a significant difference between the height of the plants grown in the two different environments. Our inconclusive results could be explained by human error, improper planting and nurturing, and environmental effects.

Coffee and plants 
Maria Avelar, Sophia Balme, Breck-Linn Patke, Emily Semans,,, 

Farmers, gardeners, or even those wishing to keep houseplants healthy often look for easy nutrients to help their plants grow faster and healthier without having to spend a high sum of money. This study focused on finding beneficial factors for plant growth from common substances one might have on hand. Our team hypothesized that the right amount of coffee grounds, which are acidic and high in nitrogen, would aid in plant growth. Nitrogen is known to be a supplemental nutrient to the growing root system, to a certain extent. To test such a claim, we sprouted radish seeds in three different concentrations to determine to what extent the nitrogenous coffee grounds would aid growth. We predicted that our 50/50 percent mixture of coffee and soil would produce the tallest sprouts of our experiment, as it would adequately supply the root system with nitrogen, though not in extreme excess. However, unfortunate consequences of our experiment setup led to the soil samples containing coffee to instead grow mold rather than the radish seeds. As such, we were unable to confirm our hypothesis of the positive relationship between plant growth and coffee ground concentration. We instead determined future experiments involving soil samples with a much lower concentration of coffee, as well as procedures that would lower the likelihood of mold sprouts.

Optimal Foraging of the Western Gray Squirrel 
Rachel Rudy 

According to optimal foraging theory, animals will optimize energy intake by eating food that is high in nutrients and provides the most energy while also requiring the least amount of time and energy for capture. Based on this idea, we hypothesized that the Western Gray Squirrel on the CSU Chico campus will forage for foods that will provide them with the most optimal energy intake. For this experiment we’ve chosen to test squirrel’s food preference by providing them with peanuts, a low nutrient legume, and blueberries, a nutrient rich fruit. For this experiment we placed a tray of 15 peanuts and 15 blueberries out each day at 8 am for a fourteen-day period and recorded how many of each were left over by 5 pm that night. After all data was collected we ran a t-test comparing the mean number of peanuts taken (2.64) and the mean number of blueberries taken (3.29). Based on the p-value (0.19), there was no significant difference in number of peanuts removed and number of blueberries removed. Therefore, we were able to conclude that squirrels will not optimize energy intake by choosing foods with a higher nutrient content.

Food Preference of CSU Chico Western Grey Squirrels 
Savanna LaPant, Mireya Limon, 

Animals tend to try and maximize their benefit by eating the more nutrient dense, more accessible food sources. The objective of this experiment was to examine food preferences between shelled/unshelled peanuts and shelled/shelled pistachios in the Western Gray squirrels on the Chico State campus.  Since peanuts are more nutrient rich and are easier to access, we hypothesized that the Western Grey squirrels will prefer to eat the unshelled peanuts in order to maximize their benefit. The experiment was carried out by placing 5 arrays array of shelled/unshelled peanuts and pistachios for a two week period. After our data was collected we can conclude that there is no statistically significant difference in nut preference (p-value 0.556). We cannot accept our hypothesis and conclude that squirrels preferred to eat the nuts with the most nutrients.

Analysis of individual density dependence of Winter Vetch based on distance from an open water source
Jessica Blankenship, Chris Del Santo, Eddie Green,, 

We studied how distance from an open water source (ex. creek, pond) affects the individual density of Winter Vetch, Vicia villosa. A small distance from an open water source will cause significant change individual density.  In order to collect data to investigate this phenomenon, individuals were located along Big Chico Creek in Upper Bidwell Park and tangent lines were measured along both 2 meter and 10 meter distances.  Individuals were counted on square meter plots and logged.  After data analysis, we concluded that there is a large discrepancy in individual density, with individuals tending to grow farther from an open water source, with statistical means of 14 individuals at 2m distance and 27 individuals at 10m distance, thus having a greater distance father away (means rounded due to inability to have partial individuals; p-value of 1.6 x 10-5).  A conclusion was reached that with regards to the Winter Vetch, individual density is highly dependent on distance from a water source, with greater density being found further from a water source.

Decomposing fruit vs. Normal soil experiment 
Mario Velazquez, Damaris Diaz, Enrique Figueroa, Alexandra Smith,,,

In our experiment, our goal as a group was to see a difference in growth with plants that had decomposing fruit added to the soil as opposed to plants grown in regular soil. We hypothesize that soil in the surrounding environment of a plant will affect its growth. To test our Hypothesis, in 4 weeks, we grew twenty radish seeds, 10 of them were grown with soil and the other 10 had both soil and banana peel. Throughout the experiment we were able to acquire 7 measurements of the plants and calculate height averages which for the plants grown in regular soil was 4.2 cm and for the ones grown in the banana peel with the soil, it came to about 3.33 cm. With that, once we calculated the degrees of freedom and p-value (0.27) we could in fact accept our null hypothesis and reject our hypothesis, saying that there is a noticeable difference in growth in plants grown with just soil as to those grown with decomposing fruit.


The effects of altered soil nutrients on Cherry Radish growth 
Melissa Diaz, Jannely Jimenez, 

In order to evaluate the effects of soil nutrients in plant growth, we researched how cherry radishes would mature when watered with different water types. We hypothesized that when altering the water used, the plant’s height would be impacted due to the nutrients absorbed from the different water types.  To test our hypothesis, 12 samples of each water type were tested accompanied with one cherry radish seed. The samples were watered every other day with their designated altered water type for two weeks total. At the end of the experiment, every sample’s height was recorded. The samples tested with salt water resulted in no sprouting seeds, which is why our results were influenced by the control group and the sugar water group. Our data suggests that a majority of the radishes were able to grow along the same height comparatively between tap water and sugar water. Given that the p-value was greater than 0.05, we cannot reject our null hypothesis. With our evaluation, we concluded that the nutritional factors in sugar water and tap water can influence growth in cherry radishes, while salt lacks those required nutrients causing the seeds to not grow.

Relationship between leaf abundance and distance from water source 
Oscar Kalfsbeek, Matthew McDonald, Daljot Singh,, 

In order for there to be significant development of plant leaves, there must be a reliable source of water; without it, the leaves can not develop. The objective of this study was to determine the effect that distance from the water source has on the plant’s leaf abundance. We hypothesize that plants closer to the water source will have more leaves per centimeter than the further away plants due to increased water availability that can be dedicated for leaf growth. Ten Vinca major plants from a near (5 meters) and far (15 meters) plot from Big Chico Creek were used as our sample.  The number of leaves per centimeter was calculated for the two plots and compared. The plants from the far plot had 8.2% greater number of leaves per centimeter than plants from the near plot, but the difference was not statistically significant (p-value = 0.27). These results did not agree with our proposed hypothesis which led to the conclusion that increased water supply does not have a significant effect on abundance.

Soil Types Affect Growth in Radish Plants 
Elizabeth Winders, Anh Thu Ma, Natalie Alba,, 

Plants need water and carbon in order to grow and plants grow better in soils that retain their moisture and are composed of carbon rich materials. Two types of soil were obtained to conduct this experiment: a peat moss soil-less mixture and an organic soil. Using radish seeds, we planted 10 seeds into each type of soil with one in each cup. Every three days the plants were watered and observations and height measurements were taken to distinguish the growth difference in the radish plants between the two soil types. Final observations showed that more radish plants were produced in the peat moss mixture and the plants that grew in the peat moss mixture grew an average of 2.3 times more in length than the radish plants that grew in the organic soil. These results showed there was a significant difference in plant growth between the soil-less mixture and organic soil (p-value= 0.003).

A Squirrel's Preference 
Shyanne Rizzo, Stephanie Ibarra, 

Optimal Foraging Theory states that the time and energy required to obtain food may determine how and what an animal chooses to eat. Our experiment was based on observing squirrel preferences of both shelled or unshelled peanuts, as well as foraging on dirt or grass. Our hypothesis was that squirrels will take more unshelled peanuts than shelled peanuts. Every morning, we set out arrays of shelled and unshelled peanuts at two different plots and would collect data every afternoon, making observations on the weather and surroundings as well. Unshelled peanuts were more preferred by squirrels, averaging about 2-3 more peanuts than shelled. Plot 2 (dirt) had a higher turnout of peanuts taken than Plot 1 (grass), nearly doubled with an average of 35. Our p-value was .551 so we cannot reject our null hypothesis, concluding that squirrels have no preference. The use of different kinds of nuts in multiple areas of different environments could change the results if this experiment was replicated.

Effects of Nitrogen Density on Radish Growth 
Ariana Rahman, Marikris Morales, Tia Peete,, 

The use of nitrogen in fertilizers is largely popular with nitrogen being a limiting nutrient to productivity in crops. Nitrogen assists in plant growth and amino acid, enzyme, and protein function. With nitrogen being an imperative nutrient in plant health, our experiment focused on the effects nitrogen on of radish growth based on different densities of nitrogen fertilizer in soil. It was hypothesized that an increased density of nitrogen fertilizer in the soil would increase radish growth. Ten samples each of 0%, 50%, and 100% nitrogen fertilizer to soil with one radish seed each were placed out for 24 days and watered. The growth of each treatment was measured and shown to have a significant difference (p-value =2.3x10^-10) Although results were significant, two treatments showed no growth and may have altered data. Our collective means for 0%, 50%, 100% nitrogen fertilizer in soil were 1.99in, 0in, and 0in respectively.

Pine tree needle acidity affects the growth of plants 
Cora Piper, Nely Lopez, Megan Farrell,, 

Pine needles acidify the under canopy which reduces the amount of competition they experience for nutrients. Our hypothesis is that the acidity of pine needles will affect the radish plants by decreasing their growth compared to radish plants watered with tap water. To study this, we planted eighteen radish seeds and watered nine of them with pine tree tea and watered the other nine with tap water. Our results showed that the average growth of radish plants is significantly slower (p=1.26 x 10^-10) when watered with pine needle tea than when watered with tap water. This indicates that pine tree needles limit the undergrowth causing the soil to be more acidic with the fall of pine needles.


BIOL 350, Rachel Schleiger,

Ecological facilitation 
Alex Willis, Bisma Khan, Theodore Lee,,  

Ecological facilitations in the environment are relationships between species that can be beneficial or harmful for a species. These interactions can be classified as mutualisms where both species benefit from one another, or commensalism where one species is neutral and the other is benefitted. To observe these relationships we tested the ecological facilitation  between Plant species Blue oak (Quercus douglasii) and poison oak (Toxicodendron diversilobum) which exhibit a commensalism relationship. But the question is how close in distance does the poison oak have to grow to receive these ecological benefits, or what is the optimal distance from the nurse plant for the other species to benefit from it.  By testing the question at hand, we collected ten samples of measuring the width of the canopy of the blue oak trees, and measuring  the distance of the poison oak plants under the canopy of the blue oak. We did this to determine whether one plant enhances the growth of the other. With the collected data we compared the average of the poison oaks distances under the Blue Oak’s canopy, to the collective average of all the Blue oaks canopy widths to determine an optimal growth distance for the poison oak. Using a one-tailed T-test to determine which plant enhances the growth of the other, we subtracted from two sets of data to find a difference between the two distances that we recorded and determined a p-value of 0.003576.  With this p-value we can conclude that the relationship of the two species is with the success of the blue oak tree, the poison oak will benefit and grow alongside the nurse plant, which in this case is the blue oak tree.

Blue Oak (Quercus douglasii) recruitment in unaltered and altered landscapes 
Bryn Copson, Dara Stroup, Rana Yousaf 

California’s oak woodlands have been dramatically reduced in extent over the past 200 years. Oak woodlands play a critical role in protecting soils from erosion and landsliding, regulating water flow in watersheds, and maintaining water quality in streams and rivers. Oak woodland ecosystems in California also have high levels of biodiversity, and there is increasing interest in oak woodland restoration projects. The objective of this study was to examine if an altered (restored) landscape either impeded or facilitated recruitment of blue oak (Quercus douglasii) seedlings compared to an unaltered landscape. It was hypothesized that the altered landscape would impede recruitment of blue oak seedlings due to soil compaction and nutrient removal by the previous plot use as an orchard. Each plot was split into a hundred 10 meter by 10 meter plots. A random number generator was used to choose 30 plots for each of the landscape types. A count of blue oak (Quercus douglasii) seedlings (height less than 30 cm) present in the 10 meter by 10 meter transect was collected. Blue Oak seedling success rates were significantly higher (p = .0114) in the altered landscape when compared to the unaltered landscape. This data demonstrates the viability of potential oak woodland restoration sites.

Decomposition Rates in Woody Plants: California Sycamore (Platanus racemosa), Valley Oak (Quercus lobata), and Manzanita (Arctostaphylos spp.)  
Cynthia Banales, Judith Celis, Rachel Libby, Jenny Salazar,,,   

The study seeks to measure the rate of decomposition of three common riparian species: California Sycamore (Platanus racemosa), Valley Oak (Quercus lobata), and Manzanita (Arctostaphylos spp.), located near Big Chico Creek in Chico, Ca. We hypothesized the manzanita woody plant will have a slower rate of decomposition due to its thick, leathery-waxy, cuticle leaves. These variations in leaf composition will define rates of decompositions in this riparian waterway, providing information about microbial and invertebrate species present, and potential for nutrient dispersal to higher order water. 25 grams of the three leaf species was collected, separated into four bags with a bag representing each week, and thus placed in Big Chico Creek. Every week, for four weeks, a bag from Platanus racemosa, Quercus lobata, Arctostaphylos spp. was removed from the creek and dried in the Lundberg Blue GO1350A laboratory heater for 24 hours at 100 degree celsius. After the drying period, the dried weight was obtained. After four weeks, the percent loss data of each species was inconclusive due to interference of soil entering the bag, creating a negative correlation. Further research would be needed by adding additional weeks of the three plant species in the creek to measure the rate of decomposition.

Blue Oak Regeneration in Upper Bidwell Park 
Jamont Strickland, Diego Arriaga, 

Blue Oak (Quercus douglassi), endemic to California, is the most abundant hardwood forest type tree in the state. Despite the fact that the Blue Oak is well adapted for California’s Mediterranean climate, it has been poorly regenerating. Oak trees are known to regenerate by advanced regeneration, where the seedling directs the majority of resources to its root system. The seedlings which constitute advance regeneration are suppressed by competition from the overstory, which results in little shoot growth. Death of overstory canopy releases seedlings, which respond with rapid growth, and regeneration results. This experiment investigates the difference between the abundance in which offspring occur whether they be within the outer canopy margin of the paternal oak, or outside of it. It was hypothesized that more seedlings would be found on the outside due to less shading therefore less competition for sunlight. In this experiment, a population of 15 Blue Oaks in Upper Bidwell Park were selected and then 6-8 transects equidistance from each adult tree and 20 meters long were made. Along each transect, we recorded the number of Blue Oak seedlings that were within and outside the canopy margin, and repeated until we had a large enough sample size for the groups. The data showed a significant correlation between the abundance of seedlings found within the dripline versus those found outside the dripline. The data collected is significant due to the fact that Oak tree regeneration is heavily dependent on advanced oak regeneration to repopulate and compete with other species.

Decomposition Rates of Deciduous Leaves 
Rachel Rudy, Miranda Olea, 

Leaf decomposition from deciduous trees into freshwater streams is an important part in nutrient cycling for aquatic communities. Stream productivity relies on the nutrients and organic matter provided by the decomposing leaves. Deciduous leaf decomposition rate varies based on environmental conditions and physiological factors such as surface area of the specific leaf type. Decomposition rates of BigLeaf Maple (Acer macrophyllum), Manzanita (Arctostahpylos spp.), and Valley Oak (Quercus lobata), were compared while each species was submerged in a local freshwater source, Big Chico Creek. Initially 80g of each species was weighed out in lab then divided into four trials, after each week one bag per species was pulled out, reweighed, and a final mass was recorded. At the end of the four-week trial the decomposition rate of each species was measured. Based on the results of the experiment there was a significant difference in decomposition rates between each species (P=0.045). The Manzanita decomposed at a significantly slower rate than the Big Leaf Maple and Valley Oak. Ultimately, leaf decomposition rates vary based on leaf composition, thickness, and overall area.

Higher content granaries influence the likelihood of robbing behavior in Sciurus griseus 
Stephanie Aguiar, Ivan Gonzalez, 

The foraging of food can be very costly to any animal species, as energy must be expelled scavenging in order to meet the nutritional demands necessary.  The western gray squirrel (Sciurus griseus) is known for robbing the granaries made by woodpeckers, as a way to expend less energy while gathering resources. The higher the granary content, the higher the probability of foraging from squirrels. When granaries have a high amount of acorn content, squirrels will tend to interact with those granaries more often. Western gray squirrels were observed at five different granaries located on CSU Chico’s campus and the One-Mile Recreation area of Bidwell Park. Estimated granary content was noted, and the number of interactions from squirrels at each granary were observed. The data demonstrates that the squirrels frequented those granaries possessing less overall content, to those with higher acorn content. It can be assumed that these results might be attributed to the possibility of the higher content granaries being visited with a higher frequency in the past.


The Abundance of Aquatic Invertebrates in Relation to Stream Flow 
Ty Hagberg, Sam Franson, Brooke Hall, 

Big Chico Creek and Butte Creek, located in Butte County California, both support healthy aquatic ecosystems. The purpose of this study is to analyze the relationship between stream flow, in CFS, and the abundance of aquatic invertebrates. This study is of importance as California has experienced one of its wettest winters on record and the data could be of value for predicting the effects of these events on aquatic biota in the future. The study was conducted over three sites at each creek with elevation being maintained between the two stream sampling areas. At all sampling sites for each stream, twenty rocks were carefully collected from the stream and the nymphs counted. It is hypothesized that due to the lower flow of Big Chico Creek, at 387 CFS compared to 890 CFS for Butte Creek, there will be a smaller population of mayfly nymphs as CFS diminishes.  The data collected from the two streams indicates that there is significance, derived from a 2-way-ANOVA statistical test with a p-value of 1.115E-12, in the relationship between stream flow and population size of mayfly nymphs. It is hypothesized that due to nutrient availability and a higher abundance of food sources in the larger stream, the mayflies have a higher carrying capacity per rock. Interestingly, Big Chico Creek was also devoid of adult caddis larger than one inch, while nearly a third of the samples at Butte Creek contained one or more. However, as Butte Creek maintains a cooler water temperature annually at the same elevation as Big Chico Creek, further studies must be conducted in order to understand the population dynamics on an annual basis.

Allelopathic effect of Lavandula and Rosmarinus officinalis on Brassica  
Jessica Coronel, Manveer Dhillon, Amandeep Dhillon,, 

Allelopathy can cause one plant to either benefit or be harmed by another plant species.  This experiment was conducted to evaluate the allelopathic potential of lavender and rosemary. The main focus was to record the germination success of the mustard seeds in the presence of the allelopathic plant.  The experiment was performed by crushing and creating an extract of the allelopathic species.  Then, the mustard seeds were planted in the presence of the allelopathic species and the control, which in this study was water.  The germination success and rate was recorded over a 3-4 week period. The control sample had a total of 3 seedlings germinate while the treatment sample did not germinate at all. This is indicative of the strong allelopathic qualities of the lavender and rosemary. It can be concluded that allelopathic species can influence the germination success of another plant.

Allelopathic effects on seed germination  
Illiana Cajias, Korina Sigala, Tristan Scott, Stephanie Barajas,,,  

Allelopathy in plants work by releasing allelochemicals that can be classified as secondary metabolites which inhibit germination of other plant species. Allelopathy in one plant can hinder the germination, growth, survival and reproduction of other organisms. This study hypothesized that allelopathic plants will slow the germination of other plants. To test this, five potential allelopathic species (walnut, sage, bay, pine, and eucalyptus) were compared to a control. Each plant was crushed and then soaked in water with filter paper overnight to allow the water to be saturated with the plants oils. Six petri dishes were prepared, one for each species in question and one for the control. Six seeds were placed in the center of the petri dish on top of the soaked filter paper, then a layer of topsoil was placed. The petri dishes were placed in a window sill where they each received equal amounts of sun. Over the course of 15 days, data was collected every 3 days where the number of germinated seeds within each petri dish. At the end of the 15 days the germination success rate was calculated for each plant. Results that Pine was the most allelopathic among our selected species. Allelopathy is an important role in the plant ecosystem because it helps in weed control and crop productivity. These allelopathic compounds within plants are a much safer and natural alternative than pesticides and herbicides and can naturally keep a balance in the plant ecosystem.

Vegetation density in relation to distance from vernal pools 
Madison Brandt, Micheal Klemm, Clinton Underwood,, 

Vernal pools are among California’s most diverse and ecologically important ecosystems, as well as one of the fastest declining habitat types in the state. These pools are home to a wide variety of both plant and animal species, many of which are state and federally listed endangered species. The species that inhabit these pools have to endure dramatic changes in conditions throughout the year, and as such the species observed within and around the pools throughout the year are subject to change. This study aims to gain a deeper understanding of the relationship between the percent cover of grasses and forbs and distance from the edge of the standing water within the vernal pools at North Table Mountain Ecological Reserve. An area of vernal pool habitat was sampled and five vernal pools were randomly selected as sample sites. At each sample site, five 0.25 meter square quadrats were randomly selected and percent cover of grasses, forbs, and bare ground was measured. Analysis run on the data collected revealed insufficient data to suggest a relationship between the distance from pools and percent cover of grasses or forbs. These results could indicate a departure of vernal pools at this location from the convention that forbs are more numerous at closer proximity to the center of vernal pools. Data collected in this survey could be used to suggest further exploration of the makeup of the vegetation in and around vernal pools at North Table Mountain Ecological Reserve.

Characteristics of storage trees used by acorn woodpeckers   
Anna Burns, Maci Meyers, Sydney Sullivan,, 

Urban environments can limit the critical resources available to wildlife. In areas more characteristic of a natural habitat, species tend to be more successful. Some respond to fluctuations in food availability by foraging for more than is needed when abundant and storing it in caches for later use. This study examined the caching behavior of the California acorn woodpecker (Melanerpes formicivorus) and was particularly interested in the trees selected for as food storage sites (granaries) in a semi-urban, naturalized park setting. The hypothesis tested was that acorn woodpeckers require a large surface area and easy access to stored food, therefore the trees selected for use as granaries would be large, oak trees with trunk circumference of at least 3m, and consistently the largest tree within 4.5m. Percent of excavated surface area of all granary trees in a 500-meter-long transect was recorded and considered a quantitative measure of acorn woodpeckers’ preference for the tree, and environment. Circumference at breast height was noted, as well as the number and size of other trees within 4.5 meters. The hypothesis was tested by running linear regressions with percent cover against the three distinct explanatory variables of granary tree circumference, number of other trees nearby, and their circumference means. Data suggested no relationship amongst the variables, resulting in rejection of the hypotheses. Additional studies might be useful toward improving understanding of acorn woodpeckers’ food caching behaviors and thus enhance management of parks and other natural corridors used by this species in the future.


Spatial patterns of Quercus lobata seedling recruitment in Bidwell Park 
He-Lo Ramirez, Mariby Cruz, Dominic Valeriote, Allison Hardeay,,, 

The Valley Oak (Quercus lobata) is a keystone species of the Sacramento Valley. Understanding its reproductive successes is important because it provides habitat and nourishment within the ecosystem. This study serves to help better understand Valley Oak spatial patterns of seedling recruitment of the Valley Oak population found in Lower and Upper Bidwell Park in Chico, CA. We hypothesized that Valley oak saplings prefer to grow near the drip line of the mature canopy cover, because this area has less competition for sunlight from the mature trees in the interior of the canopy, while being shielded from extreme heat faced in the area outside of the canopy. Thirty-eight saplings were sampled. An ANOVA indicated a significant difference amongst the means between drip line, interior, and exterior saplings (F-value = 7.530 with a p-value = .020). A Tukey's post hoc test was performed revealing that the significant difference amongst the means was due to the interaction between the interior and drip line means of seedlings (p-value of 0.02). These insights may be applied to better serve state and local municipality efforts towards rehabilitating Valley Oak populations within the Sacramento Valley.

Decomposition Rates in Woody Plants: Acer pseudoplatanus, Quercus lobata, & Arctostaphylos 
Jeanette Burdick, Tara McColm, Zachary Nelson,,   

The team collected a variety of fresh leaves from three different trees which include the California Sycamore (Acer pseudoplatanus), the Valley Oak (Quercus lobatus), and Manzanita (Arctostaphylos) with the intent on studying each of the leaves to determine their various rates of decomposition as well as the rate at which they return the nutrients that the consumed back into the ecosystem. To achieve this, 4 bags of each specimen were assembled and then placed in Big Chico Creek to initiate decomposition. A dry weight and wet weight of each leaf type was collected every week to determine the amount of decay the leaves experienced. After a total of 4 weeks, all replicates showed great signs of decay although the Manzanita showed the greatest loss in biomass. It was hypothesized that the California Sycamore leaves will decompose slower because of the trees adaptability and close exposure to water that will resist decomposition. The resulting data depicts a gradual decline of the Manzanita biomass, which may be due to the waxy layer on the outside of its leaves. This coating is an adaptation the Manzanita has developed, and is a common adaption amongst other plant types whose are native lands are prone to wildfires. This decline in biomass may be due to the fact that they are not typically found in a riparian environment. The other two specimen, the Valley Oak and California Sycamore, had a subtle decline in decomposition because of the leaves ability to hold more biomass and water in comparison to the Manzanita, thus supporting the hypothesis. Similarly, the California Sycamore and Valley Oak adapted to retain water and biomass in response to their natural riparian habitat.

Determination of Niche Partitioning in Birds Along a Portion of Big Chico Creek 
Monica Stanwick, Caylin Stanley, cstanley4@mail.csuchico 

In ecological communities where factors such as interspecific competition and predation influence species distribution, niche partitioning can facilitate the coexistence of species, ultimately preserving and furthering biodiversity. A study was conducted near a portion of Big Chico Creek to determine the extent of niche partitioning between bird species of various sizes in the surrounding vegetation. Five surveys were conducted to determine the heights in vegetation at which different sized birds frequented. When birds landed, they were recorded by size and by visual estimation of height in the vegetation. After conducting a statistical analysis, no evidence was found presenting a significant relationship between bird size and visitation height in vegetation. However, it was determined that the two most frequently sighted bird species of the study, the Acorn Woodpecker and the European Starling, showed a significant difference in their height ranges. Based on the results, it is unlikely that niche partitioning near Big Chico Creek occurs in bird species based on size and vertical height in vegetation, although further studies should be conducted to determine the extent of niche differentiation at other spatial scales.

Interspecific Exploitation by Squirrels 
Sang Pharn, Joseph Lu-na, 

In this research, we observed the interspecific exploitation of resources by California Ground Squirrels (Spermophilus beecheyi) and Western Gray Squirrels (Sciurus griseus) on the granaries of Acorn Woodpecker (Melanerpes formicivorus) through direct interspecific competition. We hypothesize that both squirrel species were equally likely to raid the Acorn Woodpecker granaries located at Lower Bidwell. In order to determine whether the two squirrel species exploit the granaries, we surveyed a specific patch of trees at the One Mile at Bidwell Park for any activities regarding these exploitations and recorded the amount of times the squirrel species raided the granaries. We then conducted a T-test in a Microsoft excel program to find out if there was any significant difference in exploitation activity between the two squirrel species. The calculations led to the discovery of non-significant results (p=0.36) regarding that the two squirrel species were equally likely to raid the Acorn Woodpecker granaries.

Foraging ecology: Giving-up densities in squirrels 
Daisy Avila, Victoria Coia, Kassandra Inzunza-Gonzalez, Rachael Simon,,, 

Behavioral responses of prey to predation risk are often mediated by vegetation structure. To assess the role of vegetation coverage in determining behavioral responses to foraging under predation risk, we used measurements of giving-up density (GUD) of vigilance behavior of Western Gray Squirrel (WGS), Sciurus griseus, across open vegetation to dense vegetation area. The GUD is the percentage of food left unconsumed that will be measured. This measure assesses when a particular food patch may become too risky that the cost of foraging outweighs any benefits. An experiment was conducted to test the GUD of WGS in open and dense vegetation areas. It’s hypothesized that the GUD will be lower in the denser vegetation areas for the WGS due to lower risk of predation. Two aluminum pie pans were set out containing 140 grams of seeds. The pans were placed near habitats of the WGS, on the Chico State campus near the creek. The pans sat out for twenty-four hours for eight days. After conducting the experiment, a linear regression graph was performed. The coefficient of determination (.21) indicates a weak relationship between the two variables.  Having a low test statistic (1.58) and a high p-value (0.26) indicates a non-significant correlation between the foraging behavior of the WSG in the open and dense vegetation area. The data does not support the hypothesis which could have been caused by the wind, people and the spot of interest. An understanding of foraging behavior in animals can lead to an understanding of forest regeneration. Many animals serve as seed dispersers and are thus essential for the propagation of tree species and essential for habitat preservation.

Niche partitioning in Solenopsis xyloni and Tapinoma sessile 
Sukhwinder Sarai and Kelly Scott, 

Ants are one of the most common insects encountered on any given day. They live in close quarters with other ant species as well as many other types of insects. The purpose of this study was to understand how these creatures utilize their environment differently than their competitors. Three different categories of food were provided for the ants grain, meat, and sugar. We observed how many ants visited each food particle and noted how long they stayed and whether they carried any food away with them. A Chi-squared test indicated a large difference in the choice of resources between the two species (P-value of <.0001 and a Chi-square value of 39.85). We believed both species would choose the same high glucose food source so we rejected our hypothesis. The ants exhibited a behavior known as resource partitioning, which is very common in the animal kingdom. They must utilize different resources because Gause’s principle states no two species can be complete competitors and coexist. Niche complementarity is another way animals avoid competition, by utilizing different parts of their habitat. This is what we predicted the ants would do, aggregate themselves apart from competitors. What we observed was a large overlap of the two species and a clear preference for meat among Solenopsis xyloni  and a preference for grains among Tapinoma sessile.

BIOL 350, Stephanie Foster,

Niche Partitioning in Birds 
Lynsey Gilreath, Miriam Ibarra, Trevor Moore and Priscilla Morales,,, 

In order to avoid interspecific competition niche partitioning is a commonly observed phenomenon in most organisms. The goal of our study was to assess if the local bird species inhabiting Bidwell Park engage in niche partitioning so that conservation efforts may focus more directly on target species. In order to begin to examine this question we surveyed an area in Bidwell Park, where we recorded the frequency at which individuals from each of the observed bird species landed on a tree along with its relative location in relation to the trunk. The hypothesis that was tested was whether different size classes of birds show distinct preferences for certain regions of the tree. Based upon our results it appears that the three size classes of birds observed do not in fact appear to be showing distinct preferences for certain regions of the trees (P=0.11). This suggests that the birds may be partitioning in some other way than by location on the branches of the trees. It is also possible that some of the species observed were not utilizing the tree as a niche and were simply landing without foraging and competing for resources. In that context some species may be able to coexist by utilizing shared tree’s region differently.

Estimating Importance Values and Variance In Tree Species 
Maison Power, Kelsey Hanson, Jacob Bomagat,, 

The relative importance of tree species in a forest is of great interest to ecologists who are attempting to understand community ecology. By understanding the community ecology, we are able to see which species are more prevalent and determine which species exhibit the most dominance. We ran an experiment at One Mile on five different tree species: Valley Oak, California Buckeye, White-leaved Manzanita, Scrub Oak and Gray Pine using the point-quarter sampling method to calculate the importance values for each species. It is hypothesized that Valley Oak and California Buckeye will have a higher importance value indicating higher dominance than Scrub Oak, White-leaved Manzanita, and Gray Pine due to higher succession rates in riparian and chaparral environments at lower elevations. Using an ANOVA, we calculated a p-value of 1.22E-13, which allows us to reject our null hypothesis, concluding that there is variance between the tree species. After calculating our importance values, we also see that our hypothesis is supported which allows us to conclude that Valley Oak and California Buckeye exhibit more dominance than Scrub Oak, White-leaved Manzanita, and Gray Pine at One Mile and that variance between tree species is present.

Giving Up Density in Western Gray Squirrels 
Jessica Shippen, Annie Enos, Teresa Levya, Breanna Tobar,,, 

The giving up density is indicative of the foraging behavior of an animal. The quantity of food that is left behind is correlated to the risks the animal faces while foraging for food instead of allocating energy towards safety, development, or reproduction. Western gray squirrels (Sciurus griseus) can be found in large numbers in many different regions.  Each of these regions have different types of pressures that act upon these squirrels, and data was collected in order to determine if the giving-up-density of Western gray squirrels is correlated with the amount of human activity around which their food source is found. We hypothesized that the giving-up-density of the Western Gray Squirrel (Sciurus griseus) is correlated to the presented risks (the presence of predators, costs of missed opportunities, and energetic costs of achieving food) of urban “low risk” areas and rural “high risk” areas. Pans of sand and sunflower seeds were placed in both urban and rural areas for 24 hours at a time. The remaining sunflower seeds were counted at the end of that time period.  These totals were used in a one-tailed t-test to determine if the giving-up density differed between rural and urban landscapes. Our t-test reflected a p-value of 0.17, which allows us to conclude that there is no significant difference in the giving-up density of the western gray ground squirrel between urban and rural settings.

Effects of Allelopathy on Germination of Radish Seeds 
Brandon Maddox, Casey Combs, Eric Madrigal, Taunya Shilling,,, 

Allelopathy is the chemical inhibition of one plant by another plant, by releasing toxins that impacts germination, growth and survival in a negative manner.  In testing our three species of plants Quercus lobata (Valley Oak), Juglans hindsii (Black Walnut), and Eucalyptus globulus (Eucalyptus) we hypothesized that the Raphanus sativus (Radish) seeds germination success should be significantly lower when exposed to the allelopathic leaves. Radish seeds were placed on filter paper soaked in the leaf litter extract and monitored for a period of seven days. There is statistical evidence that supports that the success of germination with allelopathic leaf litter is different than success of germination with control (p-value<.00001). After seeing the results, there may be another factor other than allelopathy that contributes to the success of germination of R. sativus.

Decomposition Rates in Woody Plants 
Michael Elfgen, Zach Scott, Charles Wheeler, Muhammad Khan,, 

Leaf litter from trees into freshwater creeks and streams plays a key role in the nutrient cycling of aquatic species. The purpose of this research was to study the decomposition rates of three commonly found tress along streambanks in the Butte County region – Valley Oak, Big Leaf Maple, and Manzanita leaves. These three species were collected and utilized in the experiment. Each week for a total of four weeks, several leaf filled bags submerged in Big Chico Creek were withdrawn from the water and weighed as a dry weight after drying. The new dry weight was compared to the initial measured weight to determine if or how much leaf decomposition occurred for each leaf species. The results showed that the decomposing rate of Manzanita Leaves was much faster than the decomposition rates of Big Leaf Maple and Valley Oak leaves. This could be due to the fact that the Manzanita Leaves were not as compacted in the mesh bags compared to the other two species looked at. It allowed for more water and turbulence to occur and essentially break down the leaf structure at a faster rate.          

BIOL 369, Christopher Ivey,

Possible divergence of Mimulus glaucescens populations in Butte County  
He-Lo Ramirez, Melissa McCall, Samuel Martin, Caysie Hughes, Aithne Loeblich, Angeline Jacoby, Holly Bennett, Stephen Butcher,,,, 

Edaphic adaptation can contribute to isolating barriers amongst different ecotypes, and may lead to eventual speciation. Adaptations to different soil have been witnessed in multiple Mimulus species. We investigated the possibility of local or edaphic adaptation between four different populations of Mimulus glaucescens, which grows on and off serpentine soil, within Butte County. In a greenhouse, we reciprocally transplanted the four home site seeds onto soil from each home site and recorded germination successes, rosette diameter, flowering time, reproductive effort, and dry biomass. Results were found to be composed of mixed support for edaphic adaptation, with no support for local adaptation. Seed by soil interaction displayed large p-values for all measurements save rosette diameter. However, the data may suggest divergent populations within the Mimulus glaucescens populations in Butte County.

Populations of Mimulus glaucescens experience differential success on and off serpentine soils 
Kyle Gunther, Laura Lampe, Daysi May, Evangelina Sandoval Duran,,, 

Local and edaphic adaptation can provide means for divergent evolution within species. Determining the presence one or both of these forces requires extensive testing. Serpentine soils have high levels of toxic metals, and are low in calcium, high in magnesium, thinly composed, and prone to drought and erosion. Taxa that thrive on these soils must be uniquely adapted. When found both on and off these conditions, they inspire questions about local distribution patterns and how edaphic variation influences plant evolution. Mimulus glaucescens, a regional endemic, is an example of a species found on both serpentine and non-serpentine soils. We expected serpentine and non-serpentine populations to show differential success on opposing soil types, which could suggest divergence. We conducted a greenhouse reciprocal transplant study of two serpentine and two non-serpentine populations and measured germination success, rosette diameters, flowering times, reproductive effort, and dry above-ground biomass. Using ANOVA tests to determine whether there was evidence for divergence, we observed distinct trends regarding germination and growth. Serpentine seeds experienced higher germination success and one of the serpentine sites yielded high reproductive effort and earlier flowering. All seeds displayed high reproductive effort and greater biomass on one non-serpentine site. Our results indicated differential growth success on serpentine and non-serpentine soils, but not necessarily strong support of local or edaphic adaptation.

BIOL 409, Juan Araujo Sarinana,

Investigation & Function of Gene AT3G21770 
Aarondeep Pamma, Inderveer Chahal, Raymon Dhami 

The concept of inserting transgenic proteins into a host species can offer numerous advances in biomedical research and applications. The gene AT3G21770 is one of 73 that belongs to the class III secretory peroxidase family of Arabidopsis thaliana. A transgenic variant of the gene was infiltrated with jellyfish yellow fluorescent protein (YFP), which successfully permitted visual identification of the transformed seeds under microscopic observation in which we observed our transgenic products fluoresce. We wanted to determine the location and function of the gene. Our main question of what is the function of our selected gene and where will the protein it codes for be found? Taking the independent and dependent variable into account, we formed our hypothesis, the hydrophobic nature of the C-terminus should cause our protein to be found in the vacuole, as a result of protein translation. In the vacuole our protein would help with nutrient storage within the cell or it may also help maintain turgor against the cell wall by controlling the flow of water by active transport via a potassium ion pump. The methods that we used to carry out our experiment included the construction of the YFP and the construct which was then cloned into the DNA of the PMN20 plasmid. The results of the fluoresence allows us to accept our alternate hypothesis that the protein is indeed found in the vacuole. For future work we could figure out ways to be more efficient to make this process have more conclusive results in the end.

A Transgenic Investigation of Arabidopsis thaliana Peroxidase Gene AT4G33420 Expression and Protein Localization 
Dutra A., Simonian N., Wimer L.,, 

Arabidopsis thaliana contains seventy-three class III secretory peroxidase genes, many of which serve multifunctional purposes through the plant’s physiology. Of the seventy-three, peroxidase gene AT4G33420 was used in an investigation of protein targeting. Hypotheses of the location of this gene’s expressed protein were made using bioinformatics. It was hypothesized that if the peroxidase gene was expressed, the targeted protein would be found in root tissues, and more specifically, subcellular localization of the protein would be in cell walls. In the AT4G33420 gene, native N-terminus sequencing was conserved in order to preserve signal peptide sequencing that targets insertion of the protein to the ER.  The native hydrophilic C-terminal segment was also conserved; which lacked KDEL sequencing, resulting in protein targeting to the cell wall. In order to preserve the native promoter sequencing, a procedure was developed that introduced a YFP gene tag into a selected site within our target gene. The YFP gene would be used to assess protein location using fluorescence microscopy. To prevent gene silencing, a viral promoter (CaMV35S) was introduced to Arabidopsis thaliana. It was determined through fluorescence microscopy that the protein entered the ER after expression, traveled through the Golgi apparatus, and was targeted to the cell walls; identified by the fluorescent illumination of the tagged YFP gene in root tissues.

Expected Location of AT3G21770 in Transgenic Plants 
Jacob Bomagat, Salaman Rasoli, Makenna Soudan,, 

The Peroxidase gene family (PER) has a wide variety of functions in all land plants and specifically within Arabidopsis thaliana including pathogen defense, lignification, and suberization. Because of the many functions within the large peroxidase family, it is difficult to assign a specific role to each Class III peroxidase. Using bioinformatics, the gene PER 30 (AT3G21770) was chosen and was thought to be localized in the roots. This lead to the hypothesis that the location of the protein coded by the gene AT3G21770 would be seen in the root, but more specifically in the cell wall because AT3G21770 lacks the ER retention sequence KDEL and contains hydrophilic terminal amino acids. An experiment was conducted to detect the subcellular and anatomical location of the protein coded by the gene AT3G21770. To begin, the gene was tagged with yellow fluorescent protein (YFP) using Tri-Template PCR. The fluorescently tagged gene was then cloned into the pDONR221 plasmid and finally subcloned into a second plasmid, pMN20GW. pMN20GW was used to transform Agrobacterium tumefaciens, which was then pipetted onto Arabidopsis thaliana flowers for the transformation of the developing egg/embryo to create transgenic seeds. These seeds were then harvested, plated, and allowed to germinate. Using microscopy, fluorescence was seen in the lateral root caps of the germinated seeds, concluding that the protein coded by the gene AT3G21770 is located in the roots.

Determining the function of the Peroxidase gene using Yellow Fluorescent Protein 
Jamont Strickland, Eric Tran, 

The peroxidase gene is a gene whose function is unknown, and being studied today. Recent findings suggest the gene is involved in making the protein that creates the waterproofing mechanism in the xylem, expressed primarily in the tips of the root tissue. Our experiment will be focused on establishing a correlation between peroxidase gene expression, the protein it encodes, and where it is being targeted in Arabidopsis plants. With the results of our experiment we expect to aid future scientists in understanding plant life cycles. To begin our experiment, we used tri-template PCR to insert the YFP gene into the C terminus of the peroxidase gene along with genes for kanamycin resistance. From there, we mixed our PCR products with heat shocked Escherichia coli cells and plated them onto kanamycin. The surviving colonies were mixed with heat shocked Agrobacterium tumefaciens cells and plated onto selective media. Surviving colonies were placed onto the seed pods of Arabidopsis plants. Harvested seeds were then placed onto kanamycin plates, with the surviving plants investigated under fluorescent light. If there is a correlation between peroxidase expression and xylem production, then we are expected to see fluorescence in the tip of the Arabidopsis roots. We also expect to find fluorescence in the cell wall of the plant cell based on the peroxidase protein’s hydrophobic amino acids and lack of  ER retention sequences.  With our research, we established a correlation between peroxidase expression and xylem production upon finding fluorescence in the root tips of our transgenic plants.

Expression of AT4G33420 in Arabidopsis thaliana 
Jason Saenz and Tyler Neal, 

Class III Peroxidases are extremely important to plants. Using hydrogen peroxide as an electron acceptor, it catalyzes the polymerization of lignin when targeted to the cell wall. This process transpires in the secondary cell wall in the root’s xylem cells; providing structural integrity and creates a pathway to secrete water. We formulated the question, where in the plant cell is the protein expressed from the peroxidase gene? We believe that the protein expression of our peroxidase gene is going to be located in the plant cell wall. Our peroxidase gene contains no KDEL site, meaning that the protein created will leave the endoplasmic reticulum. Also, our gene sequence contains a hydrophilic C-terminus which results in the protein most likely going to the cell wall or cell membrane of the plant cell. For our experiment, we conducted PCR on primers we produced and inserted that PCR product into a plasmid.  The PCR product contained the Yellow Fluorescence Protein which tags our protein for viewing. We then infiltrated the plant cells by drip method of our plasmid onto the flowering plants. Weeks later we collected the seeds and looked for the YFP under a microscope. From over a thousand seeds we believe that 0.3% of them to be transgenic. From our successful transformation, we are able to see YFP running down the center of the seeds root.

Protein expressed by modified gene AT3G21770 exhibits fluorescence in Arabidopsis thaliana roots 
Maddy Thompson, Katerina Arca, Haley Broadhurst,, 

In order to create genetically modified Arabidopsis thaliana seedlings, we used bioinformatics tools to choose a gene of interest and obtained the subcellular location of our class III peroxidase gene. We chose gene AT3G21770, which we hypothesized would be secreted into the cell wall. To visually analyze our experimental results, we inserted a yellow fluorescent protein (YFP) by designing primers and performing PCR on the gene of interest, followed by performing a tri-template PCR to anneal the gene fragments. Cloning of various plasmids such as pDonr and PMN20GW were done to result in the production of Agrobacterium cells.  The Agrobacterium cells contain the plasmid that was constructed molecularly, that were then inoculated onto Arabidopsis thaliana leaves, to infect the plants’ chromosomes with the selected gene.  Analysis of leaves were conducted through taking pictures of our Arabidopsis thaliana roots, where fluorescence was present in the cell, specifically in the nucleus, golgi apparatus, endoplasmic reticulum, and the cell wall.

Exploring the location and expression of Class III Secretory Peroxidase AT4G34060 within Arabidopsis thaliana through fluorescence microscopy 
Victoria Ramirez, Baldeep Sangha, 

Class III secretory peroxidase genes are plant-specific oxidoreductase that have been known to show involvement in several physiological processes in Arabidopsis thaliana. The wide variety of physiological processes that have been studied such as lignification, defense against pathogens, catabolism, and root development make it hard to classify roles for each peroxidase gene.  Our study focuses on one specific class III peroxidase gene of Arabidopsis thaliana gene family and draws a connection between peroxidase gene AT4G34060 expression and where the location of an encoded protein will be targeted in the cell.  The gene AT4G34060 was analyzed through Genevesitgator and PSORT, tagged using the YFP gene and amplified through tri-template PCR. The formed construct was then transformed into pDONR221, subcloned into an Agrobacterium plasmid known as pMN20 and then inserted into the binary vector pBLEE. After growth, Arabidopsis thaliana and Nicotiana benthamiana plants were inoculated by the Agrobacterium tumefaciens present in pBLEE to observe the subcellular location of the protein. Fluorescence microscopy helped compare plant tissues infiltrated with the AT4G34060 protein with leaves that were served as controls. Through our results, it was shown that the expression of AT4G34060 protein was visible within the xylem tissue of the apical root meristem and veins of leaves in comparison to leaves that were not infiltrated. Due to the fluorescence in these locations of the cell, the Class III secretory peroxidase gene AT4G34060 does indeed aid in processes of detoxification, lignification, defense against pathogens, catabolism, and root elongation within the plant.

BIOL 409, Mandeep Grewal,

Class III secretory peroxidase AT5G64100 subcellular location of plant cell vacuole found via comparison of control and inoculated Nicotiana tabacum leaves with YFP labeled protein 
Alyssa Bowlsby, Taeler Anderson, Savannah Epstein,, 

Arabidopsis thaliana has 73 class III secretory peroxidases with the common function of lignification in the secondary cell wall of the root vascular xylem cells, yet many of the individual functions are unknown. In order to provide evidence for the function of a single peroxidase, a subcellular location must first be determined. The N-terminus of peroxidase 69 contained a cleavage site between amino acids 23 and 24 inferring that it is a secretory protein that would be secreted outside of the nucleus. Due to the lack of an endoplasmic retention sequence and the hydrophobicity of the three terminal amino acids of the protein, it was hypothesized that peroxidase 69 (AT5G64100) would leave the endoplasmic reticulum, travel to the golgi apparatus and ultimately be found in the vacuole. This was tested by using PCR to generate coding sequence for cloning into T-DNA of pBLEE, followed by the transformation into Agrobacterium tumefaciens with pBLEE, and finally, Nicotiana tabacum leaf inoculation. Fluorescent microscopy was used to compare control and inoculated leaves and provided support to our hypothesis that the selected peroxidase subcellular location is in the vacuole. The determination of the subcellular location can be used to help deduce protein function in further research.

Subcellular Localization and Function of Peroxidase AT5G64100 in Nicotiana benthamiana With Agrobacterium tumefaciens  
Carley Corona, Nancy Martinez, and Jenny Salazar,, 

Class III peroxidases are a family of secretory peroxidase enzymes found in plant cells, and are known to support lignification in plants. Their role is to create radicals of hydrophobic lignin monomers, which bind together to form diverse lignin polymers in order to provide structural integrity and pathogen defense. In our experiment, we attempted to determine the subcellular location and function of our a peroxidase protein from the Arabidopsis thaliana genome. Bioinformatics was used to select PER 69, located at gene AT5G64100.  Our procedure included amplification, followed by isolation of our selected protein sequence using PCR. We further transformed our protein sequence into the t-DNA region of a pBLEE plasmid. The pBLEE plasmid already contained tobacco mosaic virus and a Yellow Fluorescence Protein (YFP) sequence to allow us to identify the protein. Successful transformation into Agrobacterium tumefaciens was followed by resuspension which allowed us to infiltrate the leaves of Nicotiana benthamiana. Control and infiltrated leaves were then examined under a fluorescence microscope. The presence of YFP in our predicted subcellular location successfully confirmed protein expression. Our results supported our hypothesis, indicating that expression of AT5G64100 is found inside of the cell, specifically in the vacuole and supporting our prediction that class III secretory peroxidases are involved in lignification, via precursor activity in the vacuole. Further research to confirm presence of the peroxidase and lignin in the same vacuole, since it the vacuole serve as storage compartments for nutrients and metabolites.

Investigation of AT1G05260 via Agrobacterium tumefaciens and Tobacco Mosaic Virus 
Cydney Wain, Karina Haddad, Malak Alshaibani,, 

Class III peroxidase enzymes are responsible for catalyzing lignin formation in plants. Lignin assists in structure and defense in plants. The purpose of this experiment was to determine the presence and location of a PER3 and to confirm the function of the peroxidase in the plant. All 73 Class III peroxidase genes were analyzed and a peroxidase with high mRNA content in the elongation zone of the plant was chosen. A segment of the DNA was isolated by tri-template PCR and cloned into the T-DNA portion of the pBlee plasmid vector by recombination. The pBlee plasmid contains yellow fluorescent protein (YFP) which functions in the identification and location of the gene segment. This plasmid vector was then transformed to Agrobacterium and transformed into tobacco plants. The leaves of the tobacco plant were observed using fluorescence microscopy to visualize the location of the fluorescent protein. The protein showed fluorescence in the cell wall as well as the vacuoles. These results supported the hypothesis regarding protein location. The protein contained a hydrophobic amino acid sequence at the C-terminus, indicating that the protein will be expressed in the vacuole of the plant cell. The presence of the protein in the cell wall implies it may function in lignin production.

Observation of Class III Peroxidase AT5G64100 in A. thaliana Cell Wall 
Emily Egusa, William Smith, Christopher VonTungeln,, 

Lignin is an essential component to a plant’s vascular system by creating a ring-like sheath which acts as a waterproofing mechanism to facilitate the movement of water. It is hypothesized that Class III peroxidase gene expression of Arabidopsis thaliana is responsible for the plant’s lignin production. Gene selection was based on a heat map generated by the program, Genevestigator. This mRNA accumulation map, showed that the mRNA from the gene AT5G64100 was abundant in A. thaliana vascular tissue during early development. The protein sequence of the selected gene, AT5G64100, does not contain an endoplasmic reticulum retaining sequences and the last five amino acids are not hydrophobic. Therefore, the hypothesized location of the protein in is in the cell wall. The gene was inserted into the tDNA portion of the YFP containing plasmid, pBLEE. Agrobacteria was transformed using modified pBLEE and then infiltrated into tobacco leaves.  The Agrobacteria was then used to infiltrate  tobacco leaf tissue. Fluorescence microscopy was utilized to detect any expression of YFP in the leaves. Results displayed significant fluorescence in the cell walls of the experimental plant tissue which supports our original hypothesis that the protein will be secreted into the cell wall. Using this research, future research can be done to further investigate lignin formation in plants.

Investigation of A. thaliana class III peroxidase genes 
Grace Prator, Brendan Tinsley, Melissa French,, 

It has been shown that class III peroxidase proteins in plants like Arabidopsis thaliana may play a role in lignin formation alongside their usual functions. Using various bioinformatics tools we were able to identify the gene, AT5G64100, which codes for a class III peroxidase in Arabidopsis thaliana. Is this gene’s peroxidase product involved in the polymerization of lignin? Because this peroxidase is soluble and has a hydrophobic c-terminus, we hypothesize that if we tag the protein with a fluorescent label then we will see fluorescence localized near the cell wall. We then employed cloning and subcloning techniques to transform Agrobacterium tumefaciens with pBLEE, a plasmid containing both the YFP gene, a 3xflag, and AT5G64100. A. tumefaciens was then used to infiltrate the leaves of Nicotiana benthamiana. After a week, the leaves were observed and photos were taken using fluorescent microscopy, which confirmed that the protein was being expressed in the cell wall. The presence of our peroxidase protein was also confirmed in the plant tissue by using SDS-PAGE. AT5G64100 produces a class III peroxidase that is present in the cell walls of plants. This provides a foundation for future research into the molecular mechanism of this peroxidase and the analysis of a possible target that would negate the need for harmful pollutants to be added in the paper production process.

Expression of the Protein Coded for by AT5G64100, an Arabidopsis thaliana Class III Secretory Peroxidase 
Jim Her, Usman Rehman, Vanessa Ruelas,, 

The secretory plant peroxidase family contains three different classes of peroxidases. Class I peroxidases detoxify excess hydrogen peroxide in cells. Class II peroxidases degrade soil debris. In Arabidopsis thaliana, there are 73 class III secretory peroxidases which synthesize precursors for lignin formation; lignin is a hydrophobic polymer that lines cells, such as xylem, and provides structural support to the plant. From PSORT and Genevestigator, we selected peroxidase gene 69 (AT5G64100), and tagged it with the yellow fluorescent protein (YFP), a variant of the green fluorescent protein (GFP) found in the jellyfish Aequorea victoria. We hypothesize that the protein coded for by peroxidase 69 will be expressed in the elongation and maturation zones of the root. HDEL is an endoplasm reticulum (E.R) retention peptide sequence present at the C-terminus; we hypothesize that our modified protein will be retained in the E.R. after translation. A tri-template polymerase chain reaction (TT-PCR) was performed to construct the gene AT5G64100::YFP::AT5G34060HDEL. The constructed gene was subcloned into PMN20GW (T-DNA), a cloning vector for the target gene used to transform Agrobacterium tumefaciens. The bacteria was subsequently used to transform Arabidopsis thaliana; fluorescence microscopy was performed to determine which tissue and subcellular location displayed green fluorescence. The elongation and maturation zone in the roots, and the cell wall of root cells emitted green fluorescence. Further research on expression levels of class III secretory peroxidase genes can be used to further understand the functions, regulation, and evolution of this gene family.

Engineering of Transgenic Seeds and Sub-cellular Tracking of AT1G05260 gene from Arabidopsis thaliana  
Kaylee White, Hannah Wallrich, 

The Arabidopsis thaliana is a flowering plant that houses 73 Peroxidase genes, all with a specific function. By targeting certain Peroxidase genes location, the function of each gene can be discovered. The purpose for this experiment is to find out where a specific gene, Peroxidase 3, gene AT1G05260, protein will be expressed in Arabidopsis thaliana.  Looking at the gene using PSORT, the cleavage site is between amino acid 24 and 25, which means the protein will leave the nucleus. Since the protein lacks a KDEL, it will leave the endoplasmic reticulum. The C-terminus is mainly polar amino acids, which means the protein will continue on to the cell wall. We hypothesized that the protein will be expressed outside of the cell since it is secreted outside of the cell and will function to stabilize the cell wall. To track the sub-cellular location of the gene, steps were performed, 1. Obtain and isolate our gene of interest and create gene fragments with extensions. 2. Amplify gene fragments and add in yellow fluorescent protein. 3. Clone gene using recombination and place on an E.coli host using transformation protocol, with kanamycin resistance markers. 4. Inoculate the leaves and flowers of our plants and harvest seeds. 5. Screen transgenic seeds for our gene and determine the sub-cellular location. Looking at our seeds, we see an abundance of yellow-fluorescent protein in the root elongation zone. Going forward, we can use these results to determine the function of the Peroxidase 3 gene.

Peroxidase 57, Gene AT5G17820, Fluorescence Expression in Vascular Tissue of Arabidopsis thaliana 
Lauren Davichick, Madison Hossfeld, 

Peroxidase is a stress response in plants, using redox reactions for detoxification. Other function of peroxidase includes lignification. Genetic loss is typical among all species throughout evolution. This begs the question why Arabidopsis thaliana has 73 class III secretory peroxidase genes. It is likely that every one of these genes has a specific function, which is why this plant has conserved all of these genes overtime. To determine the function of each individual gene, the location of protein expression must be identified. The purpose of this experiment is to find where PER 57, gene AT5G17820, protein is expressed in Arabidopsis thaliana. Using PSORT the cleavage site was found between amino acid 22 and 23, the gene will be cut in the nucleus and continue to the rough endoplasmic reticulum. It will go to the cell wall because the last three amino acids are mainly polar. It is hypothesized that gene AT5G1280 protein will be expressed outside the secondary cell wall. Gene PER 57 was isolated and YFP was inserted. The gene was then subcloned into the tDNA portion of PMN20 plasmid. It was then transformed into Agrobacterium and inoculated onto Arabidopsis thaliana flowers. Once seeds formed, they were harvested and place on a kanamycin agar plate. The seeds that survived were likely transgenic plants containing the transformed gene of PER 57. YFP was expressed in the vascular tissue of the transgenic plants. The next step would be to find out the specific function of this protein.

Observing cell wall subcellular location of Arabidopsis thaliana Peroxidase 69 on YFP-tagged protein structure using fluorescence microscopy 
Megan Erickson, Erica Luke, Madison Couch,, 

Peroxidases are a large family of enzymes that play a part in the plant’s defense system by generating a hydrogen peroxide production response that aids significantly in the deterrence of pathogens. There are 73 class III secretory peroxidases in Arabidopsis thaliana; the number of processes catalyzed by these peroxidases as well as the large number of their genes suggests that each individual class III peroxidase enzyme has a specialized function. This project highlights progress of identifying the specific function of our individual class III peroxidase enzyme using the model plant A. thaliana. Bioinformatics tools were used to select the AT5G64100 peroxidase gene and determine its subcellular location in the cell. The gene was expected to be expressed in the endoplasmic reticulum of cells located in the elongation zone of the root of Arabidopsis due to the presence of the ER retention signal sequence HDEL. After analysis of the amino acid sequence of peroxidases, a prediction was made that AT5G64100 would transcribe proteins that would eventually be expressed in the cells walls of the plant Nicotiana benthamiana once transfer DNA from the pMN20 plasmid had been transgenically inserted via the infectious bacterium Agrobacterium tumefaciens. The altered AT5G64100 gene contains a YFP coding sequence that causes the target peroxidase protein to fluoresce, allowing for better visualization of the location of the peroxidase. The DNA coding sequence does not have an ER retention signal HDEL on its C-terminus, allowing the protein to be placed in the bio-secretory pathway, sending it ultimately to the cell wall for secretion.

Peroxidase 69 is located in the cell wall of Arabidopsis thaliana determined using fluorescent microscopy 
Montana Loveday, Michael Xiong, Carson Hunt,, 

The general function of the 73 class III secretory peroxidases is to catalyze the formation of lignin, a structural component of plant cell walls; however, the individual function of each peroxidase is unknown. Knowing the intracellular location of peroxidase 69 will support the investigation of its individual function. This research explored what the individual function of peroxidase 69 is. Peroxidase 69 travels through the biosecretory pathway and is involved in lignification displaying fluorescence using YFP at the cell wall. We designed oligonucleotide primer sequences that could anneal the peroxidase 69 gene to use as a transcript to amplify the DNA. Homologous recombination was used to insert peroxidase 69 into pBLEE. Positive colonies containing the gene construct were identified and transformed into Agrobacterium tumefaciens. Leaves of Nicotiana benthamiana were infiltrated with Agrobacterium suspension where T-DNA from pBLEE was cut out and introduced into the plant genome. Finally, fluorescent microscopy was used to observed infiltrated leaves to identify the subcellular location of peroxidase 69. Infiltrated leaves displayed fluorescence in the cell walls, supporting our hypothesis. Further studies involving attachment of different retention signal sequences could be done to study the function of peroxidase 69 in Arabidopsis thaliana.

Peroxidase AT5G17820 is targeted to the cell wall in Arabidopsis thaliana 
Navneet Kang, Mayson Lin, Batool Al Dawood,,  

Class III peroxidases are members of a large multigene family in the plants. 73 genes were identified in the genome of Arabidopsis thaliana. Peroxidases are involved in broad range of physiological processes throughout the plant life cycle including auxin metabolism and lignin formation. However, the individual function of every peroxidase still remains unknown. By determining the location of a peroxidase in the cell, we might be able to find the potential function of the peroxidase. By using bioinformatics tools, those tools indicated the protein coding sequences from AT5G17820 contained cleavage sites, lack of KDEL sequence, and absence of hydrophobic amino acid sequences at C-terminal, thus we hypothesized that peroxidase AT5G17820 would be located in the cell wall within Arabidopsis. To test the hypothesis, we built peroxidase construct by using polymerase chain reactions to fuse yellow fluorescent protein. We eventually subcloned the construct into tumor inducing DNA vector pMN20, transformed into Agrobacterium, and then inoculated the bacteria on the flowers of Arabidopsis. Once seeds germinated, we observed the presence of peroxidases under fluorescent microscopy. The fluorescent microscopy images showed location of peroxidases residing along the root tips, which confirmed our hypothesis. Overall, the results suggested presence of peroxidase AT5G17820 in cell wall within root tips, and further attempts might be required to investigate the role of AT5G17820 in the cell wall.

Investigation & Function of Gene AT1G05260 
Rammy Sooch, Sandeep Sahota, Lucas Biggio,, 

The application of transgenic proteins into a host species offers a variety of biomedical opportunities. The gene AT1G05260 belongs to the class III secretory peroxidase family consisting of 73 genes from Arabidopsis thaliana. As part of our study, a transgenic variant of this gene was infiltrated with jellyfish YFP allowing for visual identification of our successfully transformed seeds. We asked: what is the subcellular location of AT1G05260 and what are its functions? By considering our independent variable (hydrophilic nature of the C-terminus) and the dependent variable (identification of fluorescence at various subcellular locations), we formed our hypothesis. We hypothesized that a conserved, hydrophilic C-terminus suggests that our protein will be found on the cell wall and the endoplasmic reticulum, as a result of protein translation. It will be involved in lignin formation, an essential phenolic polymer structure found in vascular plants. The methods for this experiment consisted of construction of the YFP and the construct which was cloned into the t-DNA of PMN20. This process was then followed by the Agrobacterium tumefaciens transformation and the Arabidopsis thaliana transformation. As a result of these procedures, fluorescence was observed in the cell wall of transgenic Arabidopsis thaliana plants and in the elongation zone of the root tip. The presence of lignin in high concentrations retards the absorptivity of paper, however, its removal is a highly expensive and toxic process. In terms of significance for future its removal in the plants beforehand would present an extremely efficient alternative to this process.

BIOL 409, Kristopher Blee,

Class III Peroxidase Arabidopsis thaliana gene, AT4G33420 Protein Expression in Plant Cell Wall 
Ashley Meyer, Elyn Chagolla, Ashley Addington,, 

Lignin formation is important for vascular and supportive tissues within plants, as well as aiding in the conduction of water in plant stems. Peroxidases are thought to be involved in the polymerization step of lignin biosynthesis.The peroxidase gene AT4G3320 was investigated in order to formulate our hypothesis that peroxidase class III will be distributed to the plant cell wall due to the mature protein lacking a KDEL sequence and containing three polar amino acids on the C-terminal end of the protein. Genomic DNA from Arabidopsis thaliana was isolated and a polymerase chain reaction was performed using two sets of forward and reverse primers as well as a YFP primer. DNA was amplified using PCR and analyzed in gel electrophoresis, followed by recombinational cloning by inserting the tri-template construct into pDONR221. E. coli was then transformed and plated in kanamycin containing agar. PCR and agarose gel electrophoresis were used to evaluate the transformation success. Recombinational subcloning was done by inserting the cloned pDONR221 into the pMN20W plasmid. A. tumefaciens was then transformed with the subcloned pMN20W plasmid and agarose gel electrophoresis was used to evaluate positive colonies. Transformed A. tumefaciens was inoculated to Arabidopsis thaliana plants in order to generate transgenic seeds which were plated onto kanamycin containing plates. Plants that were able to grow in the kanamycin plates would be taken for fluorescence microscopy analysis, it is expected that the cell wall of these transgenic plants fluoresce due to the recombinant peroxidase class III tagged with YFP.

Cell Targeting of Enzyme Peroxidase AT4G33420 in Arabidopsis thaliana 
Brandon Maddox, Casey Combs, Kerrick Swangler,, 

Identifying the targeted location of the peroxidase AT4G33420 enzyme in Arabidopsis thaliana can provide aid in identifying the role the enzyme has within the cell. Peroxidase enzymes are affiliated with the conversion of radical lignol molecules into the formation of lignin polymers. Lignin is a complex hydrophobic organic polymer found in xylem as structural thickenings. Our study addresses the potential location of the targeted peroxidase AT4G33420 enzyme in an A. thaliana cell. The targeted location in the plant cell is the independent variable and fluorescence detected is the dependent variable. Looking at the amino acid sequence and seeing that the last four amino acids are not KDEL we would hypothesize that the peroxidase would be targeted by the cell wall. Transformation of A. tumef flowering bodies using A. tumefaciens containing pMN20GW, a plasmid vector transformed by recombinant reporter construct, PEROX::YFP,  formed by tri-template PCR holding the native enhancer sequence. Secondary procedure involving recombinant viral promotion of peroxidase, into plasmid vector pBLEE containing DNA coding for a YFP. Transformed A. tumefaciens inoculated into lower epidermal Nicotina benthamiana leaf tissue. The results of looking at the leaf/root tissue under a microscope concluded that there was a fluorescence in the cell walls of the cells A. thaliana when compared to a controlled leaf/root cell. Results support the peroxidase AT4G33420 enzyme gets targeted to the cell walls of A. thaliana. This illustrates that peroxidase AT4G33420 is targeted to where lignin is in A. thaliana.

Peroxidase Enzyme Targeting of ATG33420 in Transformed Arabidopsis thaliana 
Cameron Divoky, Muhammad Khan, Brittany Simpson,, 

Genes for peroxidase enzymes can be found throughout all plants, animals and bacteria. The peroxidase enzymes for A. thaliana are classified as class III secretory peroxidases. Class III secretory peroxidases have several functions in a plant such as oxidizing toxic compounds, removing hydrogen peroxide and synthesizing lignin in the cell wall. The objective of the study was to monitor the intracellular targeting of ATG33420 peroxidase proteins via transformed A. thaliana plants in order to further understand the roles of peroxidases in plant cells and the mechanisms responsible for the intercellular targeting. It was hypothesized that the peroxidase 33420 protein will be targeted to the cell walls of the Arabidopsis thaliana root tips. The combination of the hydrophobic signaling peptide of ATG33420’s N-terminus coupled with the lack of KDEL at the C-terminus were reasoned to be responsible for the targeting of ATG33420’s peroxidase enzyme to the cell wall of developing root tips. After identification of ATG33420 gene using bioinformatics software, primers were designed to amplify the gene via PCR. A series of PCR amplifications followed by the cloning and sub-cloning of the peroxidase gene into various plasmids allowed for the transformation of Arabidopsis thaliana seeds. Upon examination of the plant’s cells, the presence of the yellow fluorescence protein throughout the entire plant cell indicated that the ATG33420 peroxidase protein did not exclusively target the cell wall.

Exploration of Microscopy on Transgenic Arabidopsis thaliana 
Phil McGie Hall, Jeffrey Walsh, Jessica Soria,, 

Model organism A. thaliana was used to study the formation of lignin rings in xylem. The hypothesis being tested was “Is gene AT4G33420 and its corresponding peroxidase protein PER47 involved in the formation of lignin rings in root tissue.” Gene AT4G33420 was modified to produce a YFP protein between the functional domain and the C-Terminus when translated. Roots were observed using bright field and fluorescence microscopy. Positive results would consist of fluorescent bands or spirals where xylem are located in developing root tissue. Increased fluorescence in experimental plants relative to controls would show that the YFP protein was present and the modified gene had been successfully translated.

Cellular location, and function of a single peroxidase enzyme AT5G17820 in Arabidopsis thaliana. Peroxidase is thought to be located in the xylem of the roots.  
Rachael  Simon, Sabrina Gribble, Ellen Sampsin, Lindsey Duke,,, 

Class III secretory peroxidase (PER) is one of the many types of peroxidases that are distributed in animals, plants, and microorganisms. Each PER isoenzyme has variable amino acid sequences and shows diverse expression profiles. Studies have shown that PERs participate in lignification, suberization, wound healing, etc. The question was where in the plant does the peroxidase enzyme reside?  The plant Arabidopsis thaliana PER gene AT5G17820, is a gene whose protein is targeted to the root of the plant and can be used for determining the subcellular locations of the proteins into plant cells. Based on the Genevestigator data, there was an abundance of mRNA transcripts in a particular region inside the root of the plant. This led to the hypothesis that the mature peroxidase protein will reside outside the cell wall due to the last three amino acids of the c-terminal being polar and having no KDEL sequence. KDEL refers to the abbreviations for the 4 terminal amino acids (Lysine, Aspartic acid, Glutamic acid, Leucine).The YFP coding sequence fused construct of AT5G17820 was created using tri-template PCR and cloned into the T-DNA of the Agrobacterium tumefaciens plasmid PMN20GW, Through floral inoculation, transgenic seeds of Arabidopsis were created. The infected seeds were grown and plant tissue was viewed using fluorescence microscopy. After viewing the infected plant tissue, it showed a successful transformation of the protein marked with YFP. Fluorescence was seen in transgenic roots, in the extracellular environment of the cell thus supporting the hypothesis. In order to deduce the specific gene function we will suppress the gene to see how this affects the plant.

Location of Secretory Peroxidase 47 in Arabidopsis thaliana 
Tristan Scott 

Class III secretory peroxidases are a group of enzymes that are widely speculated to support the lignification in plant cell walls. Lignin is a polymer that provides ring structures around cell walls, which provides the structural support for plants.  Research will be conducted into where the peroxidase protein (PER 47) will be secreted to within Arabidopsis thaliana. Peroxidase 47 will be expressed in heterochromatin. Upon expression it is hypothesized that a signal peptide sequences directs the Per 47 ribosomes to the rough endoplasmic reticulum. . There is a cleavage site at 29 amino acids and a lack of a KDEL sequence thus the continuation of proteins through the secretory pathway.  Peroxidase 47 proteins will then be secreted to the cell wall due to 2 of the 3 C-terminal amino acids being polar. A series of experiments were conducted designing primers to isolate the gene, constructing gene fragments containing a yellow fluorescent protein coding sequence, and creating a Tri-template pcr product.  Cloning, sub cloning, and the transformation of several plasmids were also conduct which lead ultimately to the transformation of Arabidopsis thaliana.  The results concluded that the peroxidase 47 gene is indeed secreted to the cell wall due to the presence of fluorescence along the cell wall of roots in Arabidopsis thaliana. Based on this research, this finding could prove to further investigations of other peroxidase proteins within the peroxidase family to also be secreted to the cell wall.

Functional Analysis of Class III Secretory Peroxidase AT5G17820 Gene 
Chris Viloria, Claudia Menchaca, Andrew Small,, 

Peroxidases are enzymes found in a wide variety of organisms such as plants, humans, and bacteria. They are involved in many psychological processes in plants such as the biosynthesis of lignin. Peroxidases form a large family of related enzymes that are ubiquitous in plants. There are 73 genes in the Class III Peroxidase family which have multiple tissue-specific functions. Each gene’s mechanism is yet to be determined, so our study focused on determining the individual function of the resulting gene, AT5G17820, from the plant, Arabidopsis thaliana. To determine the function of this peroxidase gene, we have to identify where the protein will be targeted into the cell as well as in the plant. We hypothesized that our protein will be targeted in the cell wall based on the absence of KDEL, the hydrophilicity of the last three amino acids at the C terminal cleavage site, and peptide cleavage occurring between the 22nd and 23rd amino acid of the N terminus cleavage site. We also hypothesized that the protein will be targeted in the root due to a high percent of expression of mRNA based on an anatomy heat map. In order to test our hypothesis, we tagged a DNA fragment which was transformed into pMN20 and later transformed into Agrobacterium. Fluorescent microscopy will allow us to visually detect the protein’s location in the cell. Inferences based on the results can then be used to either reject or support our initial hypothesis on the whereabouts and function of the AT5G17820 protein.

Cellular Targeting of Peroxidase47 in Arabidopsis thaliana 
Emma Stainton, Jennifer Morales, Guadalupe Ramirez,, 

Peroxidase (III) enzymes play an essential role in the biosynthesis of lignin- a polymer used by xylem. Protoxylem are a xylem type, that are found stacked on top of each other and upon maturation are dead and hollow. Their cell walls thicken and become ring like, allowing them to serve as “ribbed” straws. These ringed “ribs” are made of cellulose and lignin and prevent the cells from imploding as pressure is exerted on them during the uptake of water. However, the cellular location of the gene remains under question.  In this study, we observed PER47, which is coded for by ATG33420.1 and is strongly expressed in root cells of Arabidopsis thaliana. The gene sequence lead us to predict that PER47 is targeted to the lignin coated cell wall. To investigate the expected location of our protein, we transformed Arabidopsis thaliana using a bacterial plasmid containing PER47 tagged by yellow fluorescent protein (YFP). Using fluorescence, we were then able to confirm the presence of our gene in the plant cell wall.

The N and C-terminal domains of class III secretory peroxides' target the protein to the cell wall  
Stephanie Keck, Ashley Oropeza, Tara Godinez,, 

Lignin is one of the main components of the cell wall in all vascular plants. Lignin polymerization in the cell wall is dependent on the presence of class III secretory peroxidase proteins. If lignification of the cell wall is dependent on the presence of secretory peroxides', then what properties of the peroxidase protein determine its final location in the cell wall? This led to the hypothesis that the N and C-terminus determine the peroxidase protein’s involvement in the lignification of the cell wall. To test the hypothesis, molecular cloning was used for inserting the recombinant peroxidase DNA into plasmid vectors for transformation of E. coli, Agrobacterium tumefaciens, Nicotiana benthamiana (tobacco) leafs and Arabidopsis thaliana roots. Bacterial based Gateway cloning was used for generating transgenic Arabidopsis plants; while gateway compatible tobacco mosaic virus (TMV) based vectors were used for transient gene expression in tobacco leafs. With the use of fluorescence microscopy, the fluorescent peroxidase was observed in the predicted protoxylem vascular tissue of Arabidopsis roots as well as in the cell wall of the tobacco leaf cells. Both experiments support the hypothesis and conclude that the N and C-terminal domains of the peroxidase protein commits the protein to the cell wall for polymerization of lignin. Based on the current study, future research could involve the study of relative gene expression of peroxidase genes at different developmental stages of cell growth and in different cell types. 

BIOL 434, Jay Bogiatto,

Selective vs. Random Distribution of Birds in Chaparral Environment With Notes on Avian and Plant Species Diversity 
Caylin Stanley, Anton Dresler,  Jared Decker,,  

Our scientific study was conducted in order to observe the habitat partitioning of avian species in the Chaparral environment. Our mission was to determine if preference of species location was present in relation to plant diversity as well as foliage height. Our study area was located at the Big Chico Creek Ecological Reserve off of Highway 32 in Butte County. Conducting one vegetation and five avian surveys, our team collected sufficient data in order to calculate a series of diversity indexes to measure the richness of birds, vegetation, and foliage height. By inserting these derived values into further statistical test, we were able to determine if there was a relationship between height diversity and the frequency of avian visitation thus proving a selective preference among the species. Our purpose in our statistical analysis is to disprove our null hypothesis that there is no selection of foliage height as well as type of foliage by the birds of the surrounding area. To disprove this, we calculated results portraying selective utilization of plant species by birds. Among our plant species, we recorded species such as Buckbrush (Ceanothus cuneatus), Gray Pine (Pinus sabiniana), and a broad category of Grasses and Forbes. For avian species, we recorded Lazuli Bunting (Passerina amoena), California Quail (Callipepla californica), Wrentit (Chamaea fasciata), as well as many others. Our data analysis in relation to these species is ongoing and will be presented in our poster.

Habitat Utilization and Selection by Birds within a Foothill Woodland Blue Oak, with Notes on Avian and Vegetation Diversity 
Rebecca Pilakowski, Leslie Esquivel, Nikkie Werner,,  

We conducted an investigation of habitat utilization by birds within a low elevation foothill woodland blue oak ecotone located in upper Bidwell Park, Chico, Butte County, California.  Data were collected between April 10 and May 1, 2017, along 100 m transects within our 1 ha study site.  The objectives of our study were (1) to determine avian and vegetation species composition within our study area, (2) to generate a foliage height profile for our site, (3) to calculate and then assess the relationship between Shannon Diversity values (H’) for avian species, foliage height, and plant species within our habitat type, and (4) to determine whether birds are randomly or selectively using those plant species and vegetation foliage height intervals present within our study area; our null hypotheses state that there are no differences in the proportion of plant species and foliage height intervals available within our study area and the utilization of these habitat components by birds.  Our results will be presented on our poster.

Habitat Utilization and Selection by Birds of the Sacramento River Riparian Forest with Notes on Avian and Plant Diversity 
Thomas Dehoney, Sarah Smither, Jamison Sydnor, Tanner Talan,,, 

We conducted a survey of the avian community within a riparian forest habitat on a restoration site adjacent to the Sacramento River, 8 mi west of Chico, Butte County, California.  Birds were surveyed for 2 hr beginning at sunrise during the month of April 2017.  Transects were designed and surveyed for both plants and birds within our 1 ha study area.  Our goals were (1) to determine the species composition of both the avian and plant communities within are study area, (2) to calculate a foliage height profile of the vegetation present, (3) to calculate Shannon (H’) diversity values for plant species, foliage heights, and bird species to be used in a comparison with values generated within other habitats, and (4) to determine whether birds use the various plant species and foliage heights randomly or selectively; our null hypotheses being that there are no differences in the utilization of these habitat components by each bird species relative to the availability of each habitat component.  Our results, including data analysis, will be presented on our poster. 

BIOL 476, Troy Cline,

Isolation and characterization of bacteriophages isolated from raw sewage 
Jennifer Anthony, Laura Bowker, Brenda Casillas, Oscar Chavez, Rocio Clara, Stephanye Frias, Jose Gomez, Tyler Harris-Krull, Brette Heady, Carson Hunt, Douglas Lee, Montana Loveday, Erica Luke, Renee Margolin, Tonia Mubaraka, Charlotte Park, Mandeep Phagura, Baylee Russell, Ellen Sampson, Katelyn Schneider, Cathy Vue, Michael Xiong 

Bacteriophages are viruses that infect bacteria and are among the most numerous and diverse organisms on earth. Despite being critical for the discovery of many foundational principles in molecular biology, genetics, and virology, there is still much to learn about bacteriophages. Here we report the isolation and characterization of six distinct bacteriophage isolates from raw sewage collected at the Chico Water Pollution Control Plant and isolated from four different bacterial hosts (E. coli MM294, E. coli C91, Pseudomonas aeruginosa, and Serratia marcescens). We performed experiments to begin to understand some of the structural and functional properties of the phages that were isolated, including 1) whether the phages contained a lipid envelope, 2) the temperature stability of the phages, 3) the host range and attachment specificity of the phages, and 4) the burst size (ie – how many infectious phage particles are released during one round of replication) of the phage. Although all of the phages that we isolated were non-enveloped, the other parameters that we measured showed varying levels of specificity for each phage. These data highlight the great diversity that exists among bacteriophage.

BIOL 484, Mandy Banet,

Investigating Scarabaeidae Beetle Selectivity Among Native, Introduced and Exotic Mammal Dung 
Andrew Overton, Drew Nielsen, Tanner Talan,, 

Elephant conservation is shifting from zoo settings to preserves, which imitate natural settings in attempts to increase general welfare, survivorship, and birth rate of captive elephants. Adult African Elephants produce over a hundred pounds of dung daily, making research exploring natural waste management essential to the creation of such preserves. Local Scarabaeidae beetles such as the Canthon simplex and the Aphodiine Dung Beetle, Aphodius fimetarius, aid in the dispersion and removal of dung in California’s ecosystems, and could be used by the preserves as dung removal organisms. This study is investigating California Scarabaeidae beetle affinity for African Elephant dung in the presence of cow and Black-Tailed deer dung, both of which are local species.  Three pitfall traps were placed in three different locations at the Big Chico Creek Ecological Reserve. At each location, one trap was baited with cow, one with deer, and one with elephant dung. Beetle frequency was recorded at each trap and they were re-baited weekly. An ANOVA test was used to compare the frequencies of dung beetles captured at each type of dung trap. Preliminary data suggest that there is no significant difference in Scarabaeidae beetle attraction to cow, deer or elephant dung. A population of these beetles could be a cost effective, ecologically friendly means of dung removal on these large scale preserves. Further research on dung removal rates could provide compelling evidence for the use of these beetles in large scale preserve settings.

Investigation of the Factors Determining Growth Strategies of Poison Oak, Toxicodendron diversilobum, on the Big Chico Creek Ecological Reserve 
Nicole Balkow, Anton Dresler, Cody Rice,, 

Poison oak, Toxicodendron diversilobum, can exhibit a variety of growth patterns, most notably shrubs and vines. We conducted a survey of Poison Oak Communities across multiple habitats on the Big Chico Creek Ecological Reserve in order to determine whether poison oak growth pattern is correlated with environmental conditions. Surveys were conducted during a three-week period from April 13th to May 4th, 2017.  Our goals were to investigate the conclusions of previous published work that vine growth occurs at a higher proportion in riparian environments over woodland, and to explore what exact variables cause this trend and whether the growth strategies have a relation with other environments. We conducted our study along several trails on the reserve in order to make access easier, but made sure to conduct our plots out of the zone of disturbance. We added a growth type that was described in a separate publication, ground cover, and recorded information on where and how the plant grew, such as environment, slope of the hill, canopy cover, and distance to the nearest tree. Our null hypothesis was that there would be no difference in the proportion of different growth types between environments. Analysis of our data showed that there were variables that had an effect on how poison oak grows, but not exactly what we expected. Results will be displayed on the poster.

Does Flower Structure Influence Pollinator Visitation in the Big Chico Creek Ecological Reserve (BCCER)? 
Patrick Hanna, Jaclyn Lewis, Michael Spinola, Lucas Urlie,,, 

Co-evolution between pollinators and angiosperm (flowering plants) species can lead to obligate mutualism, where species traits are specialized and cause a symbiotic reliance on one another for survival. Trait expression of distinct plant structures within the flowers morphological development over time have an incentivised effect on pollinator attraction and coincide with unique characteristics of anatomical or behavioral aspects of pollinator evolution. The goal of this study is to observe whether the flower structures of selected angiosperms in the (BCCER) have an effect on pollinator attraction. We hypothesize that the flower structure will influence pollinator visitation abundance at (BCCER). A list of hypothesized generalists and specialists were generated based on the flower structure, then the individuals were haphazardly located for observation. The observations recorded all nectar extracting pollinator visits, and pollinators were identified at least down to order level. Analyses were conducted to examine differences in diversity and the numbers of pollinators that visited each type of flower.  Results will be discussed. Further studies should focus on other flower traits that may influence pollinators such as color, scent, pheromone mimicry, marked patterns and overall abundance.

Comparing habitat use of the foothill yellow-legged frog (Rana boylii) and American bullfrog (Rana catesbeianus) with respect to overlap within Big Chico Creek Ecological Reserve  
Samantha Akel, Karissa Cunningham, Loreal Matson, Cassidy Pierce,,, 

Previous observations of the American bullfrog (Rana catesbeianus) and its larva show a preference for slow flowing water usually observed in downstream locations. However, there have been many American bullfrog sightings within Big Chico Creek Ecological Reserve (BCCER), an area known to have the faster running waters associated with upstream riparian zones. This is of great concern due to possible habitat overlap with that of the foothill yellow-legged frog (Rana boylii), a species of special concern in California. Previous studies have shown a dramatic decrease in foothill yellow-legged frog populations when such habitat overlap is present. To better understand the habitat preference of the American bullfrog within and around BCCER and how it may potentially overlap and/or impact the native foothill yellow-legged frog, we completed visual encounter surveys documenting habitat preferences and species counts along three tributaries of Big Chico Creek as well as neighboring backwater channels. Preliminary results show habitat overlap at all backwater channels, with a marked absence of American bullfrog in the tributaries. However, reserve staff have observed a reduction in the amount of American bullfrog sightings along Big Chico Creek in the 2017 year. It is possible the reduced sightings are correlated with increased precipitation. Subsequent years of research will provide better understanding of this phenomenon and may provide further information needed to manage the American bullfrog within BCCER, in order to conserve habitat for the foothill yellow-legged frog.


Analysis of novel inhibitors of the GRB2 SH2 domain that decrease proliferation in chronic myeloid leukemia 
Stephanie Aguiar, Sofia Rodriguez, Tina Hanson,, 
Faculty: Carolynn Arpin, David Stachura,,

Chronic myeloid leukemia (CML) is a disease affecting the normal growth of myeloid cells in the blood and bone marrow caused by a chromosomal translocation linking the BCR and ABL1 genes together. Once transcribed, this fusion protein known as BCR-ABL, causes an over-proliferation of myeloid cells. Downstream of BCR-ABL is GRB2, an intracellular protein involved in cellular growth and differentiation. BCR-ABL binds to the SH2 domain of GRB2 and causes leukemic transformation. K562 cells are human BCR-ABL+ immortalized myelogenous leukemia cells; we tested novel SH2 antagonists (NHD215A, NHD215B, NHD292, and NHD2107) against them via a metabolic assay and found significant growth reduction after 48hrs (IC50 = 62.5uM, 31.25uM, 125uM, and 62.5uM respectively). Each antagonist was then tested on developing zebrafish embryos, which indicated that future studies on the effectiveness of these SH2 antagonists can be conducted in vivo, bringing us closer to determining safe treatments for CML.

Acoustic Analysis of Bat Echolocation Calls: Species Diversity and Seasonal Migratory Patterns 
Trevor Moore, Cameron Divoky, Aithne Loeblich, Shahroukh Mistry,,, 
Faculty: Colleen Hatfield,

California is home to 23 species of bats - almost half of all species in the country - yet few long-term acoustic monitoring programs exist. This study examines the diversity and activity patterns in three habitat types in northern California. Acoustic monitoring equipment recorded echolocation calls of bats and were analyzed using SonoBat software. Fourteen species were found across all habitats with seven being ubiquitous. The canyon bat and the fringed myotis were unique to canyon habitat while the long-legged myotis was only found at ELFS. The Yuma myotis was the most abundant ubiquitous species. BCCER had an early Spring peak. ELFS had substantial activity during the late summer and CSUF peaked during late Fall. Bats showed significant seasonal shifts in activity levels but remained present at the two lower elevation sites throughout the year.  Interesting seasonal variation suggests possible changes in prey availability in combination with shifting seasonal conditions.

mustn1a depletion negatively affects vertebrate hematopoiesis  
Brendan Tinsley, Arturo Berrun, David Stachura, 
Faculty: David Stachura,

We generated and investigated three hematopoietic-supportive stromal cell lines derived from different sites of zebrafish blood production and maintenance, which encourage the growth and differentiation of HSPCs. RNA sequencing analysis of these cell lines indicated 447 genes that were shared amongst these cell lines that were not expressed in non-hematopoietic supportive zebrafish stromal cell lines. One highly expressed transcript shared between them was mustn1a (musculoskeletal, embryonic nuclear protein 1a), one of two orthologs whose protein product is predicted to be highly related to other small nuclear proteins in the mustn1 family. In an effort to identify mustn1a’s role in hematopoiesis, we performed loss-of-function experiments by injecting morpholino (designed against mustn1a’s translational start site) into zebrafish embryos immediately post-fertilization. By utilizing transgenic zebrafish with fluorescently labeled erythroid cells, we found that mustn1a’s depletion in the developing embryo reduced erythroid cell production and negatively affected blood circulation. These results show that mustn1a may play an important role in hematopoiesis in vertebrates. Future studies of mustn1a will be aimed at analyzing the myeloid impact of reducing its transcript as well as further quantifying the erythroid results using flow cytometry. We also hope to perform overexpression experiments in the future to see how that might impact the developing zebrafish. Identifying and chemically modulating transcripts involved in making blood can be utilized clinically, allowing the expansion of blood stem cells in culture to treat a multitude of diseases such as anemia and leukemia.

kal1b reduction decreases vertebrate blood cells 
Tanya Schmidt, Matt Boice, Omar Pulido
Faculty: David Stachura,

Hematopoiesis is the process of blood development and formation of all blood cell types from hematopoietic stem cells (HSC). Zebrafish (Danio rerio) are an excellent model organism to study vertebrate hematopoiesis because they have a similar hematopoietic system to mammals containing highly conserved genetic processes for hematopoietic development and function. Methods employed were typical PCR protocol, cDNA formation via miniprep, morpholino injections, TOPO-TA cloning and sequencing. Prior studies have shown 447 gene transcripts are shared among hematopoietic supportive cells and of those 100 are overexpressed. kal1b was among those overexpressed transcripts indicating it’s implicated in hematopoiesis. We predict that by knocking out kal1b from the zebrafish transcriptome there will be a decrease in production of blood cells while overexpressing kal1b will cause an increase in blood cell production. Preliminary results indicate that kal1b transcript reduction in zebrafish significantly decreased platelet formation. Identifying genes responsible for thrombocyte production can be utilized clinically allowing in vitro culture to treat thrombocytopenia.

Meddling with microbial tubes: sheath production by Leptothrix cholodnii SP-6 
Kirstie B. Steiner, Malory O. Brown, Sandra Martell, Microbial Genetics Class (BIOL 472), Betsey Tamietti, 
Faculty: Emily Fleming,

Metal-oxidizing bacteria (MOB) such as Leptothrix-Sphaerotilus generate complex surface structures to avoid entombment within an iron or manganese crust and to remain suspended in the water column. Their cellular machinery generates a highly ordered organometallic fibrillar microtubule sheath, which they have been producing for millions if not billions of years. The microtubule sheath structure is well known but assembly machinery and regulation of microtubule sheath production is not. Determining sheath-related genes in MOB is complicated by lack of a genetic system. We developed a genetic system, obtained several sheath mutants, and determined the genes disrupted. Cells were sensitive to several antibiotics, underwent conjugation, and had transposition frequencies of 2.9 x 10-4. Several sheath mutants that either overproduced or did not produce sheaths were found. The genes associated with these mutants were linked to motility, suggesting a tie between sheath production and motility.

What is a key autotrophic enzyme doing in a metal-oxidizing heterotrophic bacterium? 
Malory Brown,
Faculty: Emily Fleming,

The metal-oxidizing bacterium Leptothrix cholodnii is considered an obligate heterotroph, but its genome contains ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the key enzyme for autotrophic carbon assimilation. Some phototrophic bacteria use RuBisCO to “mix” organic and inorganic carbon assimilation to maintain redox balance in reducing conditions, but little is known about RuBisCO activity in chemomixotrophic bacteria. L. cholodnii increases its cell yield in the presence of metals, elevated CO2 concentration, and microoxic conditions. However, an enzyme assay showed RuBisCO was inactive after 20 and 40 hours in cultures supplied with reduced manganese (Mn2+) and gassed with 7% CO2 and 10% O2. RuBisCO was also inactive in cultures grown aerobically with and without Mn2+. Future work will continue to examine the relationship between oxygen, carbon dioxide, RuBisCO activity, and metal oxidation using targeted gene mutation analysis.

Iron Oxidizers in the Classroom: Increasing Scientific Interest Through a Collaborative Ecological Study 
Molly Tuttle, 
Faculty: Emily Fleming,

The goal of this project is to both continue an existing ecological study of iron oxidizing bacteria in the environment and to increase public outreach efforts through scientific collaboration. It is no secret that there has been a decrease in STEM majors in recent years, and perhaps a lack of first-hand experience at a young age is partially to blame. Effective communication of scientific discovery to the general public is key in inspiring an interest in science to begin with. Collaboration between research labs on campus and the community schools hold a great potential for not only educating and inspiring the general public, but also serves as a novel way in which to establish and maintain ongoing scientific research. This proposed program would allow students in 6th-8th grades an opportunity to sample from the environment firsthand; learn new laboratory skills, evaluate their own samples, get a feel for “real-time science” and gain a sense of accomplishment by aiding current research. Students will be trained in methods of sampling iron oxidizing bacteria from its natural environment- measuring physical indicators and observing trends in morphology that can assist in determining regional and seasonal trends in different populations of these enigmatic organisms. In continuous sampling, done in part by the high school students, the Fleming lab can not only establish a means of consistent monitoring of the iron oxidizing bacterial populations during changes in California water events, but will be able to maintain contact and provide a continual educational influence to the community.

Mercury mobility and FeOB 
Sophia Phillips, Charles Brooke, 
Faculty: Emily Fleming,

Mercury is a potent biotoxin. The California coast is rich in both iron and mercury, and iron often transports mercury depositing it in estuarine ecosystems. Understanding interactions between mercury and iron is key to developing remediation strategies that keep mercury out of the marine ecosystem. In site surveys of a mercury contaminated marsh, iron-rich and FeOB-rich sediment mercury concentrations were lower than nearby sediment suggesting increased mercury mobility. I hypothesize biotic iron oxide production by iron oxidizing bacteria increases mercury mobility. To test this hypothesis I enriched for a marine FeOB to generate biotic iron oxides. I quantified the ability for mercury to be retained by abiotic oxides, biotic oxides, and naturally occurring oxides. I will use these data to grow FeOB under fluctuating redox conditions and monitor mercury bioavailability in the presence of biotic iron cycling.

Differential Susceptibility to Glutamate-Induced Excitotoxicity in Hippocampal and Cortical Neurons 
Tiffany Baer, William Smith,, 
Faculty: Jonathan R. Day,

Excitotoxicity is a form of neurodegeneration and is linked to stroke, epilepsy, and Alzheimer’s disease. The hippocampus is an area frequently linked to excitotoxic events due to an abundance of excitatory glutamate receptors on hippocampal pyramidal cells. Previous studies in this lab have identified a difference in response to glutamate excitotoxic insult between FVB and B6 mice hippocampal neurons. We asked if the same neuroprotection reported in B6 hippocampal neurons would be seen in B6 cortical neurons. A LIVE/DEAD assay was conducted to measure cell viability in primary cultures of cortical neurons in both strains under varying concentrations of glutamate. A significant difference was not observed in the cell viability of the B6 and FVB cortical neurons. The neuroprotection exhibited in the hippocampal cells of B6 mice is not seen in the cortical cells.

Serpentine Adaptation Gene Expression in Mimulus glaucescens  
Cody Rice, 
Faculty: Kristina Schierenbeck,

Serpentine soils; often characterized by low calcium to magnesium ratios, poor nutrient levels, and drought conditions; present a significant challenge to many plant species. Nevertheless, the significant selective pressure present has caused several adaptations in multiple linages to adapt to the harsh conditions. However, a genetic understanding of how these species have adapted is as of yet minimal. Multiple species of the clade Mimulus have evolved means of withstanding the toxic conditions of serpentine areas. One of these species, Mimulus glaucescens has the ability to be both tolerant of serpentine and non-serpentine soils, making it an excellent candidate for determining methods of serpentine adaptation. Root tips of individuals found on non-serpentine and serpentine soils were collected and total RNA was extracted from each. Illumina® RNA-seq was then used to determine overall expression levels in individuals found on serpentine sites. Using these results and previous research, four genes were selected for further study: Migut.H01178, Migut.M00950, Migut.J01730, and Migut.M01512. Quantitative real-time PCR was then employed to compare the expression of each gene from the different sites. Several of these genes demonstrated differential expression levels dependent on the soil in which the individuals were collected. These results provide greater insight into the genetic mechanisms of serpentine adaptation.

Induction Studies of PHB Depolymerase in Acidovorax and Pseudomonas 
Cassie Havens, Emily Egusa, Tara Burns, My Lo Thao, Daniel Edwards, Larry Hanne, Larry Kirk, 
Faculty: Larry Hanne, Larry Kirk, Daniel Edwards,

Biodegradable plastics are critical for a sustainable future. Polyhydroxybutyrate (PHB), polylactic acid (PLA), and co-polymers with valerate are examples of bioplastics that are partially degraded by excreted microbial enzymes in the environment. Bacteria capable of degrading PHB were previously isolated by our lab. Two of the isolates, Acidovorax wautersii and Pseudomonas alcaliphila, showed inducible production of a depolymerase in the presence of PHB. The monomer of PHB, 3-hydroxybutyrate (3HB), demonstrated partial induction of the depolymerase in Pseudomonas. Induction of the PHB depolymerase in Pseudomonas was partially repressed by the presence of glucose. A surfactant, Tween 20, was found to inhibit induction and activity of the depolymerase in Acidovorax. Both strains produce a protease that may degrade the depolymerase (at later stages of induction). Future work will focus on other strains and testing oligomeric esters of 3HB as inducing agents.

Isolation and characterization of avian influenza viruses in Northern California 
Elizabeth Bianchini, Analucia Barragan Trejo, Raymond Bogiatto, Robin Donatello, Magdalena Plancarte, Andrew Ramey, Walter Boyce,
Faculty: Troy Cline,

Recent human infections with influenza viruses of avian origin highlight the need for continued surveillance of avian influenza viruses (AIV) in waterfowl. In 2014, highly pathogenic avian influenza (HPAI) H5N8 first detected in South Korea entered North America through the Pacific Flyway, a major migratory route for waterfowl, and caused outbreaks in poultry in the United States. California’s Sacramento Valley is an important wintering site for waterfowl from which avian influenza viruses may be isolated. To better understand the risk posed by avian influenza viruses circulating in California, we collected cloacal swabs from 1,361 hunter-killed ducks across three hunting seasons at different locations in the Sacramento Valley. Sixteen waterfowl species were represented in our sampling. The presence of influenza viruses in cloacal swabs was determined by PCR for the matrix gene. The overall prevalence rate was 11.8% with diverse HA and NA subtypes represented. We observed a significantly higher positive rate in 2015-2016 (19.9%), a phenomenon that may be related to overcrowding on wetlands due to drought conditions. Northern shovelers had a statistically higher carriage rate (21.9%) relative to other species. Of particular interest, we detected HPAI H5 influenza viruses by PCR but were unable to retrieve an egg isolate. Three H7N3 isolates were obtained and, given recent human infections with H7 viruses, were characterized with respect to in vitro replication kinetics in mammalian tissue culture. All three H7 viruses were capable of replication in mammalian cell culture at levels similar to a human H1N1 virus.


Characterization of ism-1 in Hematopoietic Proliferation and Differentiation 
Arturo Berrun,
Faculty: David Stachura,

Hematopoiesis is an essential cellular process in which hematopoietic stem and progenitor cells (HSPCs) differentiate into the multitude of different cell lineages that comprise mature blood.    Hematopoiesis is a conserved evolutionary process, allowing the use of model organisms such as the zebrafish to investigate these processes. Bioinformatics data showed highly conserved transcripts within hematopoietic supporting stroma. One highly expressed transcript was ism-1, a secreted protein. To identify the role of ism-1 in hematopoiesis, we performed loss-of-function experiments. Utilizing transgenic zebrafish that have fluorescent myeloid and erythroid cells, we saw that ism-1 depletion in embryos reduced myeloid cells and negatively affected blood circulation. We also enumerated hematopoietic progenitors in vitro, which were reduced in morphant embryos. These results indicate that ism-1 is an important gene in normal vertebrate hematopoiesis. Identifying and chemically modulating transcripts involved in making blood can be utilized for the expansion of blood stem cells to treat a multitude of diseases such as anemia and leukemia.

Differential Effects of bcl-2 Overexpression in Various B Lymphocyte Populations  
Hannah Metzger, 
Faculty: David Stachura,

The purpose of this research is to further characterize the role of Bcl-2, a gene that inhibits cell death (apoptosis), by analyzing its expression and functional outcomes in different populations of B cells.  We will use fluorescence-activated cell sorting (FACS) to isolate B cells from distinct populations. From these, we will determine normal levels of BCL-2 expression using a western blot, and the degree of apoptosis using AnnexinV. We will also overexpress bcl-2 in some B cell populations using a vector consisting of bcl-2 driven by the IgM2 promoter and enhancer, and a fluorescent marker (DsRed).We will use FACS on the resulting transgenic line to isolate B cells to determine the levels of BCL-2 and apoptosis. This novel transgenic line could serve as a unique model that can be; a) mated with other transgenic fish to create new models for B cell leukemia or; b) used as a tool for immortalizing B cells to further characterize their development process and transcriptional properties.

Changes in male northern pintail (Anas acuta) courtship choices in the context of sex ratio, female availability, and hunter pressure 
Stephanie Foster, 
Faculty: Don Miller,

Animals must make choices in engaging in behaviors that are potentially risky or costly. In northern pintails, males may decide not to engage in courtship flights if many other males are competing in the same flights, if hunters are present, and if it’s late enough in the season. Between October and February of 2010-2011 and 2011-2012, I observed male courtship flights, pintail sex ratios, pintail pairing status, and hunter harvested sex ratios, and performed regressions to test for relationships. My results indicate that there was a relationship between courtship flight size and both female availability and time of year, and not with sex ratio or hunter harvest. This may mean that courtship flights are only costly near the end of the season when acquiring resources for migration may be more important than potentially pairing with a female.