Event Title

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.

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Apr 9th, 10:00 AM Apr 9th, 11:30 AM

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.
http://cornerstone.lib.mnsu.edu/urs/2012/poster-session-A/33