Event Title

Effects of Mechanical Wounding on the Expression of Pea Leaf Lipoxygenase Isoforms

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

Theresa Salerno

Mentor's Department

Chemistry and Geology

Mentor's College

Science, Engineering and Technology

Description

Lipoxygenases (LOX) are enzymes that catalyze the addition of oxygen to polyunsaturated fatty acids. In plants, the hydroperoxides contribute to defense against foreign pathogens. Previous work has implicated the role of the LOX enzyme. Genetic studies have shown several pea LOX isoforms exist, but only LOX N2 has been implicated in the pathogenesis response. A better understanding of LOX isoforms that are induced in these responses would be valuable in providing pesticide alternatives. This research was to examine how wounding affected the expression of LOX1: PS1, LOX1:PS7 and LOX g. Three sets of pea plants were grown in a controlled growth chamber at 22 degrees with 12 hours of light. Four leaves on seventeen day-old plants were wounded with a hemostat and then were frozen in liquid nitrogen 0, 3, 6, 12 and 24 hours after wounding. Controls (non-wounded leaves) were also collected and frozen for each time point. RNA was isolated from the frozen leaves using RNeasy Plant Mini Kit (Qiagen). The quality and quantity of each RNA sample was assessed spectrophotometrically. The RNA samples were then converted to cDNA in reverse transcription using High Capacity cDNA RT Kit (Life Technologies). The subsequent cDNAs were then used for qPCR. The delta delta Ct method was used to assess relative expression using EF-1alpha as the endogenous control. The experiment was repeated twice. The data was compiled to give averages and standard deviations. All the isoforms tested were significantly induced with wounding with maximal expression twelve hours after wounding.

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Apr 16th, 10:00 AM Apr 16th, 12:00 PM

Effects of Mechanical Wounding on the Expression of Pea Leaf Lipoxygenase Isoforms

CSU Ballroom

Lipoxygenases (LOX) are enzymes that catalyze the addition of oxygen to polyunsaturated fatty acids. In plants, the hydroperoxides contribute to defense against foreign pathogens. Previous work has implicated the role of the LOX enzyme. Genetic studies have shown several pea LOX isoforms exist, but only LOX N2 has been implicated in the pathogenesis response. A better understanding of LOX isoforms that are induced in these responses would be valuable in providing pesticide alternatives. This research was to examine how wounding affected the expression of LOX1: PS1, LOX1:PS7 and LOX g. Three sets of pea plants were grown in a controlled growth chamber at 22 degrees with 12 hours of light. Four leaves on seventeen day-old plants were wounded with a hemostat and then were frozen in liquid nitrogen 0, 3, 6, 12 and 24 hours after wounding. Controls (non-wounded leaves) were also collected and frozen for each time point. RNA was isolated from the frozen leaves using RNeasy Plant Mini Kit (Qiagen). The quality and quantity of each RNA sample was assessed spectrophotometrically. The RNA samples were then converted to cDNA in reverse transcription using High Capacity cDNA RT Kit (Life Technologies). The subsequent cDNAs were then used for qPCR. The delta delta Ct method was used to assess relative expression using EF-1alpha as the endogenous control. The experiment was repeated twice. The data was compiled to give averages and standard deviations. All the isoforms tested were significantly induced with wounding with maximal expression twelve hours after wounding.

Recommended Citation

Frolov, Briana. "Effects of Mechanical Wounding on the Expression of Pea Leaf Lipoxygenase Isoforms." Undergraduate Research Symposium, Mankato, MN, April 16, 2013.
http://cornerstone.lib.mnsu.edu/urs/2013/poster-session-A/26