Event Title

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.

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Apr 20th, 10:00 AM Apr 20th, 11:30 AM

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.
http://cornerstone.lib.mnsu.edu/urs/2015/poster_session_A/35