Time Course of the Wounding Effect on Lipoxygenase Expression in Soybean Leaves
Location
CSU Ballroom
Start Date
9-4-2012 10:00 AM
End Date
9-4-2012 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
Lipoxygenases (LOXs) are enzymes that catalyze the addition of molecular oxygen to unsaturated fatty acids to form hydroperoxide products. Soybean plants have several LOX isoenzymes or different proteins that catalyze the same reaction. Soybean seeds contain at least three LOX isoenzymes while at least six different isoenzymes are in the vegetative tissue. Expression of some LOX isoenzymes increases after mechanical wounding in soybean plants. The objective of this project was to explore the time course of the effect of wounding on the expression of Lox mRNAs. Plants were wounded at the bifoliate stage; one leaf was wounded while the other was used as a systemic leaf. Both leaves were harvested separately at 3½, 6 and 24 hours after wounding. Leaves were also harvested from control plants, which were not wounded. RNA was isolated from the samples using the RNeasy Plant Minikit from Qiagen. RNA quantities and quality were assessed by measuring sample absorbancies at 260 and 280nm. cDNAs were prepared using a High- Capacity cDNA Reverse Transcription Kit from Applied Biosystems. The Quantitative Polymerase Chain Reaction experiment was done on a Step One Plus Real-Time PCR system from Applied Biosystems using SYBR Green as the fluorescent indicator. Wounded leaves showed a significant increase in the expression of Lox 7 mRNA 3½ hours after wounding. This level was maintained 6 hours after wounding, but significantly increased again 24 hours after wounding. Systemic leaves did not show a significant increase in Lox 7 expression until 24 hours after wounding.
Time Course of the Wounding Effect on Lipoxygenase Expression in Soybean Leaves
CSU Ballroom
Lipoxygenases (LOXs) are enzymes that catalyze the addition of molecular oxygen to unsaturated fatty acids to form hydroperoxide products. Soybean plants have several LOX isoenzymes or different proteins that catalyze the same reaction. Soybean seeds contain at least three LOX isoenzymes while at least six different isoenzymes are in the vegetative tissue. Expression of some LOX isoenzymes increases after mechanical wounding in soybean plants. The objective of this project was to explore the time course of the effect of wounding on the expression of Lox mRNAs. Plants were wounded at the bifoliate stage; one leaf was wounded while the other was used as a systemic leaf. Both leaves were harvested separately at 3½, 6 and 24 hours after wounding. Leaves were also harvested from control plants, which were not wounded. RNA was isolated from the samples using the RNeasy Plant Minikit from Qiagen. RNA quantities and quality were assessed by measuring sample absorbancies at 260 and 280nm. cDNAs were prepared using a High- Capacity cDNA Reverse Transcription Kit from Applied Biosystems. The Quantitative Polymerase Chain Reaction experiment was done on a Step One Plus Real-Time PCR system from Applied Biosystems using SYBR Green as the fluorescent indicator. Wounded leaves showed a significant increase in the expression of Lox 7 mRNA 3½ hours after wounding. This level was maintained 6 hours after wounding, but significantly increased again 24 hours after wounding. Systemic leaves did not show a significant increase in Lox 7 expression until 24 hours after wounding.
Recommended Citation
Serem, Eva. "Time Course of the Wounding Effect on Lipoxygenase Expression in Soybean Leaves." Undergraduate Research Symposium, Mankato, MN, April 9, 2012.
https://cornerstone.lib.mnsu.edu/urs/2012/poster-session-A/33
