2D Electrophoresis Analysis of Environmentally Stressed Soybean Plants
Location
CSU Ballroom
Start Date
18-4-2016 10:00 AM
End Date
18-4-2016 11:30 AM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
James Rife
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
Soybeans are economically important to Minnesota with a projected harvest of eight million acres in 2015, the third highest state. Biological and environmental stresses can reduce yield, impacting Minnesota’s economy. By understanding response mechanisms to stressors in specific organs of the plants, improvements in yield can be made. Sample plants were treated for five hours with cold, drought, and salt stress. Cold stressed plants were exposed to 4°C, drought stressed plants had their bare roots exposed to air, and salt stressed plants had their roots soaked in a 0.3M NaCl solution. Leaf and root samples were collected for each group and analyzed using two-dimensional gel electrophoresis to detect changes in protein expression. Proteins were extracted using trichloroacetic acid and acetone prior to being redissolved in a 9M Urea, 1% CHAPS, 1% DTT, and 1% ampholyte solution for isoelectric focusing. The proteins were then analyzed in two dimensions. The first dimension was ran on IPG Ready Strip gels from BioRad with a pH gradient from 3 to 10. The second dimension was ran on Criterion XT Bis-Tris gels from BioRad. A BioRad Imager was used to compare intensities of protein spots in the samples. Proteins were identified on the gel and the differences in protein expression for each stressor were compared to the control plant. These changes in protein expression in response to stress provide insight into the plant’s stress response mechanism.
2D Electrophoresis Analysis of Environmentally Stressed Soybean Plants
CSU Ballroom
Soybeans are economically important to Minnesota with a projected harvest of eight million acres in 2015, the third highest state. Biological and environmental stresses can reduce yield, impacting Minnesota’s economy. By understanding response mechanisms to stressors in specific organs of the plants, improvements in yield can be made. Sample plants were treated for five hours with cold, drought, and salt stress. Cold stressed plants were exposed to 4°C, drought stressed plants had their bare roots exposed to air, and salt stressed plants had their roots soaked in a 0.3M NaCl solution. Leaf and root samples were collected for each group and analyzed using two-dimensional gel electrophoresis to detect changes in protein expression. Proteins were extracted using trichloroacetic acid and acetone prior to being redissolved in a 9M Urea, 1% CHAPS, 1% DTT, and 1% ampholyte solution for isoelectric focusing. The proteins were then analyzed in two dimensions. The first dimension was ran on IPG Ready Strip gels from BioRad with a pH gradient from 3 to 10. The second dimension was ran on Criterion XT Bis-Tris gels from BioRad. A BioRad Imager was used to compare intensities of protein spots in the samples. Proteins were identified on the gel and the differences in protein expression for each stressor were compared to the control plant. These changes in protein expression in response to stress provide insight into the plant’s stress response mechanism.
Recommended Citation
Colakovic, Ryan. "2D Electrophoresis Analysis of Environmentally Stressed Soybean Plants." Undergraduate Research Symposium, Mankato, MN, April 18, 2016.
https://cornerstone.lib.mnsu.edu/urs/2016/poster-session-A/33
