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.
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.
https://cornerstone.lib.mnsu.edu/urs/2010/poster-session-A/4
