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Effects of Caffeine and Ibuprofen on the Growth of Arab

Kyle Butzine, Jasmine Crafton, and Dr. Catherine Chan

University of Wisconsin – Whitewater, Department of Biological Sciences and Chemistry

Introduction• Pharmaceuticals and personal care products (PPCPs) are often

released into the environment, even after wastewater treatment (e.g., application of biosolids and wastewater runoff)

• Two commonly consumed pharmaceuticals are caffeine and ibuprofen.

• The effects of caffeine and ibuprofen in the environment have not been well studied.

• We test the effects of caffeine and ibuprofen on various growth aspects of a model land plant, Arabidopsis thaliana, at different life stages.

• The goal is to examine how these drugs affect plant growth and their effective concentrations.

Ibuprofen

MethodsGermination Experiment

• Grow A. thaliana seeds in standard nutrient agar media enriched with various concentrations of caffeine and ibuprofen.

Early Growth Experiment

• Grow A. thaliana seeds in regular nutrient media.

• Transplant seedlings into caffeine- and ibuprofen-containing media when plants have reached the 4 leaf growth stage..

• Compare primary root length of plants transplanted into caffeine and ibuprofen media to plants grown in regular nutrient media.

• Record the length of time it takes for roots, stem, 2 leaves, 4 leaves, and 6 leaves to emerge.

Later Stage Growth Experiment

Continuous Exposure

• Grow A. thaliana in regular nutrient media until the 4 leaf developmental stage is reached.

• Transfer plants into soil and divide plants into control and experimental group.

• Water control group with water and the experimental with water supplemented with 20 mM of calcium chloride.

• Water every 3 days.

Direct Application

• Separate flower clusters and leaves into control and experimental groups.

• Apply a set volume of ibuprofen or caffeine at different concentrations on control and experimental flowers and leaves. Repeat the same with solvent control.

• Repeat every 3 days and record differences between the control and experimental groups.

AcknowledgementsMerck/ AAAS Summer Undergraduate Research Program

WiscAMP

College of Letters and Sciences

University of Wisconsin Whitewater Undergraduate Research Program

Figure 1: Average % of seeds that germinated in different concentrations of ibuprofen within 317 hours (about 13 days). Without ibuprofen, 89% of seeds germinated. Error bars are calculated based on 95% confidence. As concentration of ibuprofen increases, a statistically significant percentage of seeds that germinate decreases.

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Leaf Emergence with Caffeine

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Leaf Emergence with Ibuprofen

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Figure 3: Average time for appearance of different plant organs of plants grown in various concentrations of ibuprofen. Plants exposed to 10 ppm of ibuprofen did not grow 4 leaves or 6 leaves within the experimental time frame. Plants exposed to 10 PPM of Ibuprofen show a statistically significant delay of all organs.

Figure 2: Average % of seeds that germinated in different concentrations of caffeine within 317 hours (about 13 days). Without caffeine, 78% of seeds germinated. Error bars are based on 95% confidence. No differences in germination between the presence and absence of caffeine or exposure to different concentrations of caffeine are observed.

Figure 6: Average amount of primary root growth after plants were exposed to various caffeine concentrations during the 4 leaf stage for 24 hours. Statistically significant differences are seen at 100 ppm caffeine. Preliminary data suggests that 2 leaf stage is unaffected by concentrations up to 1 ppm and 6 leaf is unaffected up to 100 ppm caffeine.

Germination Results

Caffeine

Figure 4: Average time for appearance of different plant organs of plants grown in various concentrations of caffeine. Caffeine appears to not significantly delay organ emergence.

Figure 5: Average amount of primary root growth after plants were exposed to various ibuprofen concentrations during the 4 leaf stage for 24 hours. Plants exposed to 10 ppm and 100 ppm of ibuprofen have a significant decrease in root length. Preliminary data suggests that 2 and 4 leaf stages plants have similar responses toward ibuprofen.

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Figure 7: Plants grown in 10 ppm and 100 ppm ibuprofen showed lateral

root growth, as oppose to the normal downward growth aligned with the

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Early Growth ResultsIbuprofen Caffeine