Expression of Lipoxygenase (LOX) 1 Ps: 5 and LOX G in Pea Leaves after Mechanical Wounding
Location
CSU 253/4/5
Start Date
4-4-2011 11:00 AM
End Date
4-4-2011 12:30 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
Lipoxygenase (LOX) enzymes are a class of catalysts that vitally contribute to plant defenses against environmental and pathogenic stressors. Several LOX isoforms like LOX 1 ps: 1, LOX 1 ps: 5, LOX 2, LOX 3, LOX 4, LOX G, and LOX 7 have been identified at the DNA level in pea tissues. Previous studies have shown an increase in total lipoxygenase activity in wounded pea plants after the plant had been exposed to biotic stressors. Very few studies have shown differences in the relative expression of individual LOX isoenzymes. In this present work, two real time polymerse chain (qPCR) methods were designed to quantitatively monitor LOX ps:5 and LOX G expression in wounded pea leaves. RNA was isolated from pea leaves using the RNeasy Plant Mini Kit (Qiagen). The quantity and quality of the RNA samples were assessed spectrophotometrically. The RNAs were reverse transcribed using random hexamers and a high capacity cDNA reverse transcription kit (Applied Biosystems). Primers and probes were designed using known sequences and Primer Express software ( Applied Biosystems). Efficiency curves showed that the qPCR designs were successful. The expression of the LOX isoenzymes were measured in pea leaves at 0, 12 and 24 hours after mechanical wounding. While LOX 3 expression increased significantly compared to the unwounded control, LOX 1 ps:5 did not.
Expression of Lipoxygenase (LOX) 1 Ps: 5 and LOX G in Pea Leaves after Mechanical Wounding
CSU 253/4/5
Lipoxygenase (LOX) enzymes are a class of catalysts that vitally contribute to plant defenses against environmental and pathogenic stressors. Several LOX isoforms like LOX 1 ps: 1, LOX 1 ps: 5, LOX 2, LOX 3, LOX 4, LOX G, and LOX 7 have been identified at the DNA level in pea tissues. Previous studies have shown an increase in total lipoxygenase activity in wounded pea plants after the plant had been exposed to biotic stressors. Very few studies have shown differences in the relative expression of individual LOX isoenzymes. In this present work, two real time polymerse chain (qPCR) methods were designed to quantitatively monitor LOX ps:5 and LOX G expression in wounded pea leaves. RNA was isolated from pea leaves using the RNeasy Plant Mini Kit (Qiagen). The quantity and quality of the RNA samples were assessed spectrophotometrically. The RNAs were reverse transcribed using random hexamers and a high capacity cDNA reverse transcription kit (Applied Biosystems). Primers and probes were designed using known sequences and Primer Express software ( Applied Biosystems). Efficiency curves showed that the qPCR designs were successful. The expression of the LOX isoenzymes were measured in pea leaves at 0, 12 and 24 hours after mechanical wounding. While LOX 3 expression increased significantly compared to the unwounded control, LOX 1 ps:5 did not.
Recommended Citation
Tambi, Sandra. "Expression of Lipoxygenase (LOX) 1 Ps: 5 and LOX G in Pea Leaves after Mechanical Wounding." Undergraduate Research Symposium, Mankato, MN, April 4, 2011.
https://cornerstone.lib.mnsu.edu/urs/2011/poster-session-B/23
