Effects of Mechanical Wounding and Methyl Jasmonate Treatment of Pea Lipoxygenase Isoforms
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
Theresa Salerno
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
Lipoxygenases (LOXs) are important enzymes for development, senescence, and responses to various stresses. In a plant wound defense mechanism, increased expressions of pea LOXs were identified along with the release of signal molecules like methyl jasmonate (MeJA). The effects of mechanical wounding and MeJA treatment on the quantitative expression levels of different LOX isoenzymes have not been well studied. Recent research has shown wound-induced elevations of LOX-g and LOX-N3 mRNA levels in pea leaves. The first phase of this research studied another pea isoenzyme LOX 1: PS 7 along with LOX-g and LOX-N3. Gene specific primers and probes were designed by the Primer Express Software (Applied Biosystems) and used to amplify LOX 1: PS 7 by Real-Time quantitative polymerase chain reaction (qPCR). Its expression also increased after mechanical wounding. In the second phase, the effect of MeJA treatment on LOX-g, LOX-N3, and LOX 1: PS 7 isoform expression was investigated. MeJA-treated pea leaves and their controls were frozen in liquid nitrogen after 3, 6, 12, and 24 hours of treatment. Their RNAs were isolated using RNeasy Plant Mini Kit (Qiagen), and a spectrophotometer was used to measure amounts and purity. The reverse transcription (RT) technique was used to convert RNA samples to the corresponding cDNA molecules. The qPCR results were normalized using endogenous EF1α, and RQ (relative quantities) of three different LOX isoforms were obtained relative to untreated controls. MeJA also increased expression of LOX 1: PS 7.
Effects of Mechanical Wounding and Methyl Jasmonate Treatment of Pea Lipoxygenase Isoforms
CSU Ballroom
Lipoxygenases (LOXs) are important enzymes for development, senescence, and responses to various stresses. In a plant wound defense mechanism, increased expressions of pea LOXs were identified along with the release of signal molecules like methyl jasmonate (MeJA). The effects of mechanical wounding and MeJA treatment on the quantitative expression levels of different LOX isoenzymes have not been well studied. Recent research has shown wound-induced elevations of LOX-g and LOX-N3 mRNA levels in pea leaves. The first phase of this research studied another pea isoenzyme LOX 1: PS 7 along with LOX-g and LOX-N3. Gene specific primers and probes were designed by the Primer Express Software (Applied Biosystems) and used to amplify LOX 1: PS 7 by Real-Time quantitative polymerase chain reaction (qPCR). Its expression also increased after mechanical wounding. In the second phase, the effect of MeJA treatment on LOX-g, LOX-N3, and LOX 1: PS 7 isoform expression was investigated. MeJA-treated pea leaves and their controls were frozen in liquid nitrogen after 3, 6, 12, and 24 hours of treatment. Their RNAs were isolated using RNeasy Plant Mini Kit (Qiagen), and a spectrophotometer was used to measure amounts and purity. The reverse transcription (RT) technique was used to convert RNA samples to the corresponding cDNA molecules. The qPCR results were normalized using endogenous EF1α, and RQ (relative quantities) of three different LOX isoforms were obtained relative to untreated controls. MeJA also increased expression of LOX 1: PS 7.
Recommended Citation
Cho, Young Hee. "Effects of Mechanical Wounding and Methyl Jasmonate Treatment of Pea Lipoxygenase Isoforms." Undergraduate Research Symposium, Mankato, MN, April 9, 2012.
https://cornerstone.lib.mnsu.edu/urs/2012/poster-session-A/35
