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 4°C

To prepare Samples for Filtered run

use a 0.45µM 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

0.45µM 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.