Generation of PVY Coat Protein siRNAs in Transgenic Potatoes Resistant to PVY

Kent F. McCue, Grisel Ponciano, David R. Rockhold, Jonathan L. Whitworth, Stewart M. Gray, Yuriy Fofanov, William R. Belknap

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Global climate change has the potential to quickly alter the distribution and profile of crop pests and pathogens. Ready tools for mobilization of resistance into elite crops will be part of an integrated strategy to maintain agricultural productivity. Engineered virus resistance through expression of coat protein is well established. The mechanisms of virus suppression by small RNAs (sRNAs) have only recently been elucidated. Understanding the role of sRNAs in gene regulation and development is rapidly evolving. High throughput sequencing (HTS) can generate large data sets of sRNA sequences which can be analyzed to recognize the abundant sRNAs that result from transcript degradation and to identify the regulatory micro RNAs (miRNA) involved in gene regulation and small inhibitory RNAs (siRNA) that confer virus resistance. Transgenic Solanum tuberosum that are glass house or field grown and that express the potato virus Y coat protein (PVY-CP) inverted hairpin RNA (ihRNA) from the 409s promoter with GBSS6 intron as a spacer exhibited resistance to PVY. Southern blots of genomic DNA indicated one or two copies of the transgene and analysis of HTS data of the small RNA population indicated high levels of siRNA production from the transgenic hairpin construct. Up to 47 of the top 200 most frequent sRNA reads were attributable to the PVY-CP transgenic construct in Ranger Russet line #5. This combination of transgene and transcription strength is functional for constructing efficient resistance cassettes for crop protection. Up to 95 of the top 200 sRNA sequences were 21 nucleotides in length with as many as 19 sequences corresponding to known miRNA transcripts. Additional endogenous sRNAs with homology to resistance gene analogs may be indicative of additional or complimentary resistance mechanisms produced in concert with the PVY siRNAs arising from PVY-CP RNA transcription.

Original languageEnglish (US)
Pages (from-to)374-383
Number of pages10
JournalAmerican Journal of Potato Research
Volume89
Issue number5
DOIs
StatePublished - Oct 2012
Externally publishedYes

Fingerprint

coat proteins
genetically modified organisms
potatoes
RNA
Potato virus Y
microRNA
transgenes
transcription (genetics)
nucleotide sequences
plant pests
viruses
genes
messenger RNA
resistance mechanisms
plant protection
Solanum tuberosum
Southern blotting
introns
nucleotides
promoter regions

Keywords

  • 409s
  • High throughput sequencing
  • ihRNA
  • miRNA
  • pBINPLUS/ARS
  • sRNA

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

Cite this

McCue, K. F., Ponciano, G., Rockhold, D. R., Whitworth, J. L., Gray, S. M., Fofanov, Y., & Belknap, W. R. (2012). Generation of PVY Coat Protein siRNAs in Transgenic Potatoes Resistant to PVY. American Journal of Potato Research, 89(5), 374-383. https://doi.org/10.1007/s12230-012-9257-0

Generation of PVY Coat Protein siRNAs in Transgenic Potatoes Resistant to PVY. / McCue, Kent F.; Ponciano, Grisel; Rockhold, David R.; Whitworth, Jonathan L.; Gray, Stewart M.; Fofanov, Yuriy; Belknap, William R.

In: American Journal of Potato Research, Vol. 89, No. 5, 10.2012, p. 374-383.

Research output: Contribution to journalArticle

McCue, KF, Ponciano, G, Rockhold, DR, Whitworth, JL, Gray, SM, Fofanov, Y & Belknap, WR 2012, 'Generation of PVY Coat Protein siRNAs in Transgenic Potatoes Resistant to PVY', American Journal of Potato Research, vol. 89, no. 5, pp. 374-383. https://doi.org/10.1007/s12230-012-9257-0
McCue, Kent F. ; Ponciano, Grisel ; Rockhold, David R. ; Whitworth, Jonathan L. ; Gray, Stewart M. ; Fofanov, Yuriy ; Belknap, William R. / Generation of PVY Coat Protein siRNAs in Transgenic Potatoes Resistant to PVY. In: American Journal of Potato Research. 2012 ; Vol. 89, No. 5. pp. 374-383.
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