Transcript UWW poster Final[1]

Mineral Accumulation of a Model Plant, Arabidopsis
thaliana, After Exposure to Caffeine and Ibuprofen
Kyle Butzine and Dr. Catherine Chan
[email protected] and [email protected]
University of Wisconsin – Whitewater
Root Growth Results
Introduction
• High volume pharmaceuticals are excreted by humans and
sometimes disposed through the sewer system.
Figure 1
•Terrestrial plants can be exposed to these pharmaceuticals through
flooding and biosolid application.
Figure 2
1cm
•Preliminary results suggest two frequently used drugs, caffeine and
ibuprofen, have negative effects on plant growth at 100 ppm and 10
ppm, respectively.
• We tested the synergistic effects of applying both caffeine and
ibuprofen at the same time to a model land plant, Arabidopsis thaliana.
•We monitored the growth of Arabidopsis during drug exposure, and
quantified mineral accumulation by Inductively Coupled Plasma
Optical Emission Spectroscopy (ICP-OES).
•Our goal is to correlate phenotypic effects of drug exposure to
differences in mineral accumulation.
Figure 1 & Figure 2: Root
growth after 24 hours of drug
exposure. T0 indicates initial
root length, and T24 indicates
root length after 24 hours
exposure to 10 ppm caffeine +
10 ppm ibuprofen. Plants
exposed to caffeine and
ibuprofen grew significantly
less than control plants.
Significant difference was
calculated based on 95%
confidence.
1cm
T0
T0
T24
T24
Control
10 ppm Caffeine +
10 ppm Ibuprofen
Methods
1.2
•Germinate seeds in standard plant agar media.
1
Ratios (treated/untreated)
•Microwave digest dried plant tissue with a mixture of 50% nitric acid, 30%
hydrogen peroxide, and 20% water.
1 ppm Caffeine +
1 ppm Ibuprofen
0.8
0.6
10 ppm Caffeine +
1 ppm Ibuprofen
0.4
10 ppm Caffeine +
10 ppm Ibuprofen
0.2
0
Aluminum
Microwave Digestion Parameters
Stage
1
2
3
Power
(W)
1600
1600
1600
Ramp Time
(Min)
5:00
5:00
20:00
PSI
400
400
400
Temp.
70
100
180
Hold Time
(Min)
5:00
5:00
10:00
• Analyze samples for macro nutrients (Ca, Mg, Na, K, S, P) and micro
nutrients (Al, B, Co, Cr, Cu, Fe, Mn, Mo, Ni, V, Zn) by ICP-OES.
•Arabidopsis exposed to select
pharmaceuticals may have altered physiology
and mineral nutrient content, even when there
is no observable phenotypic effects.
•Applying caffeine and ibuprofen
simultaneously does not result in enhanced
toxicity in Arabidopsis.
ICP-OES Results
Significantly Affected Elements
•Transplant seedlings into media with varying concentrations of caffeine and
ibuprofen and monitor growth.
Conclusion
•Aluminum and Chromium levels significantly
decrease when Arabidopsis are exposed to as
low as 1 ppm caffeine + 1 ppm ibuprofen.
Chromium
Elements
Figure 3
Significance
• Caffeine and ibuprofen may negatively affect
plant growth.
Caffeine and ibuprofen generally exist at
concentrations below 1 ppm in surface waters
but their concentrations are possibly higher in
the biosolids that are applied to agricultural land.
•Crop plants with significantly lower Cr levels
may negatively impact human and animal
health.
Chromium is believed to enhance glucose
metabolism in humans by enhancing insulin
receptor sensitivity and increasing translocation
of glucose transporter to the cell membrane.
Figure 3: Elements that were significantly different between plants treated with
various combinations of caffeine and ibuprofen and control plants. A ratio of 1
indicates no change in the accumulation of a particular mineral between
treated and untreated plants. Aluminium and chromium were present at
significantly lower concentrations in caffeine- and ibuprofen-treated plants.
Significant difference
was determined by Student’s t-test. (N=3,p<.05)
Wavelength Parameters for ICP-OES (nm)
Table 1
Table 1: The average ratio and
standard error between untreated
plants and plants treated with 10 ppm
caffeine + 10 ppm ibuprofen for
macro elements. No macro element
concentrations were significantly
different between untreated plants
and treated plants. (N=3,p<.05)
Mo
202.031
Zn
213.857
Co
228.616
Ni
231.604
Fe
238.204
Mn
257.61
Cr
267.716
Cu
324.752
Table 2
Al
396.153
Sr
407.771
Ba
455.403
Mg
285.213
Ca
317.933
V
292.402
Na
589.592
K
766.49
Sr
180.669
P
213.617
B
249.772
Element
Al
B
Ba
Co
Cr
Cu
Fe
Mn
Mo
Ni
Sr
V
Zn
Average Ratio
0.774
0.636
1.761
3.446
0.558
0.859
1.122
0.928
1.243
1.365
1.209
0.793
1.344
Element
Ca
K
Mg
Na
P
S
Average Ratio
1.291
1.302
1.368
1.470
1.698
1.333
+/- Standard Error
0.048
0.265
1.279
2.121
0.011
0.084
0.207
0.281
0.413
0.566
0.242
0.426
0.272
+/- Standard Error
0.159
0.381
0.302
0.392
0.489
0.450
Table 2: The average ratio
and standard error between
untreated plants and plants
treated with 10 ppm caffeine
+ 10 ppm ibuprofen for
microelements. Aluminum
and chromium had
significantly different
concentrations between
untreated plants and treated
plants. (N=3,p<.05)
Adapted from http://lpi.oregonstate.edu/infocenter/minerals/chromium
Acknowledgements
Merck/ AAAS Undergraduate Science Research Program
University of Wisconsin Whitewater Undergraduate Research Program
College of Letters and Sciences, Dean Mary Pinkerton
Dr. John Ejnik