Event Title

The Effect of Wounding on the Expression of MicroRNAs in Peas

Location

CSU Ballroom

Start Date

18-4-2016 10:00 AM

End Date

18-4-2016 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

Legumes and other crops are constantly challenged by abiotic and biotic stresses that decrease average yields by more than 50%. Previous studies have shown that microRNAs play crucial roles in plant defense mechanisms in response to stress. When wheat plants were wounded, decreased levels of miR-169 and miR-167 and increased levels of miR-319 were observed. In another wounding study with wild tobacco, both miR-167 and mi-RNA 319 levels were increased. This study investigated the effect of wounding on the expression of miR-167 and miR-319 in pea leaves. Little marvel pea seeds were planted in soil and placed in a growth chamber. After 14 days, leaves in one set of pots were wounded by a hemostat and leaves in a second set of pots were not treated. Leaves were harvested at 0, 4, 8, 12, and 24 hours after mechanical wounding and immediately frozen in liquid nitrogen. Small RNAs were isolated using a modification of the mirVana Kit procedure. Absorbance values at 260 and 280 nm were used to assess acceptable quality and quantity of the purified small RNAs. DNA copies of the isolated RNA were made by reverse transcription using the Taqman MicroRNA RT Kit. qPCR methods were designed to amplify the cDNA and monitor the expression of miR-167 and mi-319. Efficiency curves showed that the qPCR designs were successful. The initial studies have shown that levels of expression of miR-319 and miR-167 were up-regulated at 4 and 8 hours but not 12 hours after wounding compared to the control pea leaf samples.

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Apr 18th, 10:00 AM Apr 18th, 11:30 AM

The Effect of Wounding on the Expression of MicroRNAs in Peas

CSU Ballroom

Legumes and other crops are constantly challenged by abiotic and biotic stresses that decrease average yields by more than 50%. Previous studies have shown that microRNAs play crucial roles in plant defense mechanisms in response to stress. When wheat plants were wounded, decreased levels of miR-169 and miR-167 and increased levels of miR-319 were observed. In another wounding study with wild tobacco, both miR-167 and mi-RNA 319 levels were increased. This study investigated the effect of wounding on the expression of miR-167 and miR-319 in pea leaves. Little marvel pea seeds were planted in soil and placed in a growth chamber. After 14 days, leaves in one set of pots were wounded by a hemostat and leaves in a second set of pots were not treated. Leaves were harvested at 0, 4, 8, 12, and 24 hours after mechanical wounding and immediately frozen in liquid nitrogen. Small RNAs were isolated using a modification of the mirVana Kit procedure. Absorbance values at 260 and 280 nm were used to assess acceptable quality and quantity of the purified small RNAs. DNA copies of the isolated RNA were made by reverse transcription using the Taqman MicroRNA RT Kit. qPCR methods were designed to amplify the cDNA and monitor the expression of miR-167 and mi-319. Efficiency curves showed that the qPCR designs were successful. The initial studies have shown that levels of expression of miR-319 and miR-167 were up-regulated at 4 and 8 hours but not 12 hours after wounding compared to the control pea leaf samples.

Recommended Citation

Tachago Kamdom, Estelle Sonya. "The Effect of Wounding on the Expression of MicroRNAs in Peas." Undergraduate Research Symposium, Mankato, MN, April 18, 2016.
http://cornerstone.lib.mnsu.edu/urs/2016/poster-session-A/37