Two-Dimensional Gel Electrophoresis Analysis of Soybean Leaf Proteins
Location
CSU Ballroom
Start Date
16-4-2013 10:00 AM
End Date
16-4-2013 12:00 PM
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
Lipoxygenase (LOX) enzymes play a significant role in plant defense mechanisms along with growth, development, nitrogen storage and senescence. These enzymes catalyze the oxygenation of polyunsaturated fatty acids. Soybeans have numerous different LOX enzymes. When plants are wounded, the octadecanoid pathway is stimulated and thus higher expression of some LOX isoenzymes are seen. Methyl jasmonate (MeJA), a volatile wound signal in plants, has been shown to increase the expression of some LOX isoenzymes. This study investigated LOX isoenzyme levels in soy bean leaves at the bifoliate stage in response to mechanical wounding and (MeJA) treatment using two-dimensional gel electrophoresis. Soybeans were grown in an environmental chamber. Plants were treated at the bifoliate stage. Wounded plants had one leaf crimped with a hemostat. The other unwounded leaf was treated as a systemic leaf. MeJA treated plants were exposed to MeJA vapor in a sealed aquarium. After 24 hours i) wounded leaves, ii) systemic leaves, iii) methyl jasmonate treated leaves and iv) leaves from control plants were collected. Proteins were extracted using a procedure adapted from Natarajan et. al (Analytical Biochemistry 2005, 342, 214-220). Samples were electrofocused in a Bio-Rad Protean IEF cell using IPG ready strips for separation by isoelectric points. Then the strips were embedded on top of a SDS-PAGE gel for separation according to protein size. The LOX isoenzyme expression on the sample leaves were compared to the control leafs, which were left untreated and unwounded using a gel scanner.
Two-Dimensional Gel Electrophoresis Analysis of Soybean Leaf Proteins
CSU Ballroom
Lipoxygenase (LOX) enzymes play a significant role in plant defense mechanisms along with growth, development, nitrogen storage and senescence. These enzymes catalyze the oxygenation of polyunsaturated fatty acids. Soybeans have numerous different LOX enzymes. When plants are wounded, the octadecanoid pathway is stimulated and thus higher expression of some LOX isoenzymes are seen. Methyl jasmonate (MeJA), a volatile wound signal in plants, has been shown to increase the expression of some LOX isoenzymes. This study investigated LOX isoenzyme levels in soy bean leaves at the bifoliate stage in response to mechanical wounding and (MeJA) treatment using two-dimensional gel electrophoresis. Soybeans were grown in an environmental chamber. Plants were treated at the bifoliate stage. Wounded plants had one leaf crimped with a hemostat. The other unwounded leaf was treated as a systemic leaf. MeJA treated plants were exposed to MeJA vapor in a sealed aquarium. After 24 hours i) wounded leaves, ii) systemic leaves, iii) methyl jasmonate treated leaves and iv) leaves from control plants were collected. Proteins were extracted using a procedure adapted from Natarajan et. al (Analytical Biochemistry 2005, 342, 214-220). Samples were electrofocused in a Bio-Rad Protean IEF cell using IPG ready strips for separation by isoelectric points. Then the strips were embedded on top of a SDS-PAGE gel for separation according to protein size. The LOX isoenzyme expression on the sample leaves were compared to the control leafs, which were left untreated and unwounded using a gel scanner.
Recommended Citation
Subedi, Paurakh. "Two-Dimensional Gel Electrophoresis Analysis of Soybean Leaf Proteins." Undergraduate Research Symposium, Mankato, MN, April 16, 2013.
https://cornerstone.lib.mnsu.edu/urs/2013/poster-session-A/39
