The Effect of Mechanical Wounding on the Expression of MicroRNAs in Pea Leaves
Location
CSU Ballroom
Start Date
20-4-2015 10:00 AM
End Date
20-4-2015 11:30 AM
Student's Major
Chemistry and Geology
Student's College
Science, Engineering and Technology
Mentor's Name
Theresa Salerno
Mentor's Email Address
theresa.salerno@mnsu.edu
Mentor's Department
Chemistry and Geology
Mentor's College
Science, Engineering and Technology
Description
MicroRNAs (miRNAs) are important regulators of protein expression in both plants and animals. Recently microRNAs have been found to be involved in plant stress responses in cotton under drought conditions. Little is known about the effect of mechanical wounding on microRNA expression. In this study the effects of mechanical wounding on the expression of two different miRNAs in pea ( .) leaves, MiR-167 and MiR-408, were measured. Computer-based algorithms were used to confirm a very probable match between MiR-167 and the auxin response factor mRNA and between MiR-408 and the 3’ end of plastocyanin mRNA. In this study pea (Little Marvel) seeds were grown for fifteen days. Plant tissues were collected at 0, 6 and 12 hours after wounding and quickly frozen in liquid nitrogen. Small RNAs were then extracted using the MirVana miRNA Isolation Kit. The quantities and qualities of RNA and miRNA quality were confirmed using the Nanodrop Lite spectrophotometer readings at 260 and 280nm. For the analyses poly A tails were added to the microRNAs followed by reverse transcription of the modified RNAs. Quantitation was done using a specific forward primer and a universal reverse primer. The expression relative to a small nuclear RNA U6 was then determined on a Step One Plus qPCR instrument using the ΔΔCt method. It is expected that the expression of both MiR-408 and MiR-167 will be changed. This research will contribute to the understanding of the role of miRNAs in biotic and abiotic stress responses of plants.
The Effect of Mechanical Wounding on the Expression of MicroRNAs in Pea Leaves
CSU Ballroom
MicroRNAs (miRNAs) are important regulators of protein expression in both plants and animals. Recently microRNAs have been found to be involved in plant stress responses in cotton under drought conditions. Little is known about the effect of mechanical wounding on microRNA expression. In this study the effects of mechanical wounding on the expression of two different miRNAs in pea ( .) leaves, MiR-167 and MiR-408, were measured. Computer-based algorithms were used to confirm a very probable match between MiR-167 and the auxin response factor mRNA and between MiR-408 and the 3’ end of plastocyanin mRNA. In this study pea (Little Marvel) seeds were grown for fifteen days. Plant tissues were collected at 0, 6 and 12 hours after wounding and quickly frozen in liquid nitrogen. Small RNAs were then extracted using the MirVana miRNA Isolation Kit. The quantities and qualities of RNA and miRNA quality were confirmed using the Nanodrop Lite spectrophotometer readings at 260 and 280nm. For the analyses poly A tails were added to the microRNAs followed by reverse transcription of the modified RNAs. Quantitation was done using a specific forward primer and a universal reverse primer. The expression relative to a small nuclear RNA U6 was then determined on a Step One Plus qPCR instrument using the ΔΔCt method. It is expected that the expression of both MiR-408 and MiR-167 will be changed. This research will contribute to the understanding of the role of miRNAs in biotic and abiotic stress responses of plants.
Recommended Citation
Vinje, Samuel. "The Effect of Mechanical Wounding on the Expression of MicroRNAs in Pea Leaves." Undergraduate Research Symposium, Mankato, MN, April 20, 2015.
https://cornerstone.lib.mnsu.edu/urs/2015/poster_session_A/35
