Event Title

Expression of Lipoxygenase Isoforms in Wounded Pea Leaves

Location

CSU 253/4/5

Start Date

5-4-2010 10:00 AM

End Date

5-4-2010 12:00 PM

Student's Major

Chemistry and Geology

Student's College

Science, Engineering and Technology

Mentor's Name

Terry Salerno

Mentor's Department

Chemistry and Geology

Mentor's College

Science, Engineering and Technology

Description

Lipoxygenases (LOX) are a group of enzymes that are widely distributed in plants. There are at least five different LOX enzymes in peas. Little is known about their specific roles, although some change in amount at different stages of development. Mechanical wounding of pea leaves has shown that general LOX activity increases with peaks of activity observed at 6 hours and 24 hours after wounding. None of these studies measured individual LOX isoenzymes, and none used a very quantitative method to measure the expression of mRNAs. The purpose of this research was to measure the change in expression of LOX 1 and LOX 3 mRNAS in wounded pea leaves. The control and wounded plant leaves were harvested at 0, 3, 6, 12, and 24 hours after wounding. For each time point, the tissues from four plants (8 wounded leaves) were combined and quickly frozen in liquid nitrogen. Total RNAs from each duplicate sample were isolated using the RNeasy Plant Mini Kit (Invitrogen). The RNAs were made into copy DNAs using reverse transcriptase and random primers. Real time polymerase chain reaction (qPCR) primer and probes were designed and were used to measure differences in the levels of LOX 1 and 3 relative to a housekeeping gene (EF-1 ). Initial studies have shown significant increases in the expression of both LOX isoforms twelve hours after wounding.

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Apr 5th, 10:00 AM Apr 5th, 12:00 PM

Expression of Lipoxygenase Isoforms in Wounded Pea Leaves

CSU 253/4/5

Lipoxygenases (LOX) are a group of enzymes that are widely distributed in plants. There are at least five different LOX enzymes in peas. Little is known about their specific roles, although some change in amount at different stages of development. Mechanical wounding of pea leaves has shown that general LOX activity increases with peaks of activity observed at 6 hours and 24 hours after wounding. None of these studies measured individual LOX isoenzymes, and none used a very quantitative method to measure the expression of mRNAs. The purpose of this research was to measure the change in expression of LOX 1 and LOX 3 mRNAS in wounded pea leaves. The control and wounded plant leaves were harvested at 0, 3, 6, 12, and 24 hours after wounding. For each time point, the tissues from four plants (8 wounded leaves) were combined and quickly frozen in liquid nitrogen. Total RNAs from each duplicate sample were isolated using the RNeasy Plant Mini Kit (Invitrogen). The RNAs were made into copy DNAs using reverse transcriptase and random primers. Real time polymerase chain reaction (qPCR) primer and probes were designed and were used to measure differences in the levels of LOX 1 and 3 relative to a housekeeping gene (EF-1 ). Initial studies have shown significant increases in the expression of both LOX isoforms twelve hours after wounding.

Recommended Citation

Obidiegwu, Ifedi. "Expression of Lipoxygenase Isoforms in Wounded Pea Leaves." Undergraduate Research Symposium, Mankato, MN, April 5, 2010.
http://cornerstone.lib.mnsu.edu/urs/2010/poster-session-A/4