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United States Department of Agriculture

Agricultural Research Service

Research Project: MOLECULAR GENETIC TOOLS FOR POTATO AND FRUIT TREE IMPROVEMENT Title: Resistance to PVY using siRNA in transgenic potatoes(Abstract)

Authors
item McCue, Kent
item Rockhold, David
item Ponciano, Grisel
item Gray, Stewart
item Whitworth, Jonathan
item Fofanov, Yuriy -
item Belknap, William

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 27, 2012
Publication Date: February 8, 2013
Citation: Mc Cue, K.F., Rockhold, D.R., Ponciano, G.P., Gray, S.M., Whitworth, J.L., Fofanov, Y., Belknap, W.R. 2013. Resistance to PVY using siRNA in transgenic potatoes(Abstract). American Journal of Potato Research. 90(2):p138-139.

Technical Abstract: Virus resistance by expression of viral coat protein (CP) is well documented. The role of small RNAs (sRNA) in virus suppression has only more recently begun to be elucidated. Many sRNAs are involved in gene regulation and referred to as micro RNA (miRNA). Taking advantage of the cellular mechanism to produce biologically active sRNA sequences, we can express RNA that targets the cell’s natural machinery to produce small inhibitory RNA (siRNA) conferring virus resistance. Transgenic potatoes were generated expressing the potato virus Y (PVY) CP in an inverted hairpin RNA (ihRNA)-forming construct driven by the Solanum bulbocastanum 409s promoter. These plants exhibited resistance to PVY in glass house studies using PVYNTN and PVYO as inocula and in field studies using naturally occurring PVYNTN and PVYO. Deep sequencing the sRNAs revealed high levels of siRNA production. Up to 47 of the top 200 most abundant sRNA reads were attributable to the PVY-CP transgene. The 409s promoter drives strong RNA production and the PVY-CP ihRNA construct using the granule bound starch synthase intron as a spacer resulted in efficient hairpin formation and siRNA processing conferring resistance to PVY in transgenic potatoes. This combination of transcription strength and transgene is suitable for constructing efficient resistance cassettes for crop protection.

Last Modified: 12/19/2014
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