Methods

Abstract and Literature Cited

After germination, grow seedlings
distilled water and cultivated in a
greenhouse area for eight days with
14 hour daylight per day
Prepare Pb and Pb-
EDTA solutions for
all plant treatments
After 6 days of growth,
grow the seedlings in the
solutions of Pb and Pb-
EDTA for 8 days
Grind up plant material
with mortar and pestle
Prepare GR Assay Buffer,
GR Sample Buffer, and GR
Glutathione Reductase
Prepare Samples by follow-
ing Tissue Homogenate in
assay protocol and centri-
fuge for 15 minutes at 4C
To prepare Samples for Filtered run
use a 0.45M filter syringe; store so-
lutions on ice when not in use
Set the specphotometer at 340 nm and record
absorbance once every minute for 5 minutes to 5
obtain data points for each sample tested
Conclusion
Cayman Chemical Glutathione Reductase Assay Kit
Adjustable pipettes and a repeat pipettor
HPLC grade water
A plate reader capable of reading absorbance at 340 nm or a specphotometer
Sunflower Plant Material
0.45M filter syringe
Centrifuge
Mortar and Pestle
Based on the results of the experiment, the hypothesis was supported. The hy-
pothesis stated that less oxidative stress would be induced on plants treated with
EDTA. All cultivars treated with EDTA demonstrated higher levels of glutathione re-
ductase activity. Glutathione reductase plays a key role in protecting plants from oxi-
dative stress, and so EDTA did reduce the amount of oxidative stress on a plant.
However, in some cultivars the starting absorbance value was above 1.2 and so those
results should not be as highly regarded. The filtered run provided better results for
the dry plant material, but for the live Arabidopsis plant tissue it destroyed the en-
zymes within the tissue. For future research, the data demonstrates live tissue
should not be filtered, but dry tissue provided better and clearer results when fil-
tered. To further prove the hypothesis, it is recommended that more trials using the
assay should be done to demonstrate truly accurate results. EDTA both assists in up
taking more metal and increases glutathione reductase activity to protect the plant
against oxidative stress. Phytoextraction is a cost efficient method in removing met-
als from the soil and chelating agents have proven to make phytoextraction an even
more effective method. The Sunflower has excellent tolerance to oxidative stress
with the assistance of EDTA, and also is a biofuel crop. The oil obtained from the
sunflower can produce profits while also removing metals from the ground.