Event Title

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.

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Apr 4th, 11:00 AM Apr 4th, 12:30 PM

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.
http://cornerstone.lib.mnsu.edu/urs/2011/poster-session-B/17