Using Q-PCR To Measure Effects of Nitrogen Supplementation on the Expression of Lipoxyenase Isoenzymes in Soybeans
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
James Rife
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
Lipoxygenases (LOXs) have been described as a group of enzymes that add oxygen to fatty acids containing a cis,cis-1,4-pentadiene bond system. Soybeans have been shown to have four different LOXs in the seed and at least five different vegetative LOXs. Several roles have been proposed for the LOXs including lipid mobilization, nitrogen storage and wound response. The purpose of this project was to investigate the possible role of LOXs as nitrogen storage proteins by monitoring the effect of nitrogen supplementation on the levels of the mRNAs coding for different LOXs. Soybean plants were grown in a controlled environment. Plants were watered with a modified Hoagland‘s solution containing minimal nitrogen. Control plants received only this basic solution while experimental plants were watered with this minimal solution supplemented with NH4NO3. Leaves and roots were harvested at the bifoliate stage. RNA was isolated from the tissue samples using an RNeasy Plant Minikit from Qiagen. cDNA copies of the mRNAs were made using a High Capacity Reverse Transcription kit from Applied Biosystems. Changes in the relative levels of LOX 5, LOX 7 and LOX 9 mRNAs were measured by the Quantitative Polymerase Chain Reaction (Q-PCR) using a Power SYBR Green mix from Applied Biosystems.
Using Q-PCR To Measure Effects of Nitrogen Supplementation on the Expression of Lipoxyenase Isoenzymes in Soybeans
CSU 253/4/5
Lipoxygenases (LOXs) have been described as a group of enzymes that add oxygen to fatty acids containing a cis,cis-1,4-pentadiene bond system. Soybeans have been shown to have four different LOXs in the seed and at least five different vegetative LOXs. Several roles have been proposed for the LOXs including lipid mobilization, nitrogen storage and wound response. The purpose of this project was to investigate the possible role of LOXs as nitrogen storage proteins by monitoring the effect of nitrogen supplementation on the levels of the mRNAs coding for different LOXs. Soybean plants were grown in a controlled environment. Plants were watered with a modified Hoagland‘s solution containing minimal nitrogen. Control plants received only this basic solution while experimental plants were watered with this minimal solution supplemented with NH4NO3. Leaves and roots were harvested at the bifoliate stage. RNA was isolated from the tissue samples using an RNeasy Plant Minikit from Qiagen. cDNA copies of the mRNAs were made using a High Capacity Reverse Transcription kit from Applied Biosystems. Changes in the relative levels of LOX 5, LOX 7 and LOX 9 mRNAs were measured by the Quantitative Polymerase Chain Reaction (Q-PCR) using a Power SYBR Green mix from Applied Biosystems.
Recommended Citation
Lee, Hyuck Jin. "Using Q-PCR To Measure Effects of Nitrogen Supplementation on the Expression of Lipoxyenase Isoenzymes in Soybeans." Undergraduate Research Symposium, Mankato, MN, April 4, 2011.
https://cornerstone.lib.mnsu.edu/urs/2011/poster-session-B/17
