Isolation of CP-PVY-Specific siRNA from PVY-Infected Plants of Solanum tuberosum

Abstract

Tools for activating crop resistance to viruses are now becoming part of a comprehensive plant protection
 strategy. Artificial resistance to viruses through expression of the viral envelope protein in transgenic
 plants is fairly well understood. An urgent issue is the study of small RNAs involved in the protective mechanisms
 of RNA interference against viruses. Understanding the role of short interfering RNA (siRNA) in the
 regulation and shutdown of genes is important. The proteinase accessory component (HC-Pro), a multifunctional
 suppressor protein synthesized by the potato virus Y, is able to neutralize S. tuberosum plant defenses
 by trapping siRNA and removing them from the RNA interference process, thereby causing systemic infection
 of the host plant. Protein liquid chromatography combined with high performance sequencing can help
 recognize the large number of small RNAs resulting from viral RNA degradation and identify 21–23 bp.
 siRNA from PVY-infected S. tuberosum plants. The HC-Pro/siRNA nucleoprotein complex was detected in
 chromatographic fractions using antibodies against HC-Pro, Southern-blot indicated the presence of small
 RNAs in the complex, and analysis of data from deep sequencing of the small RNA population determined a
 specificity of 21–23 bp. siRNA to the envelope protein of the PVY virus. The research results can be applied
 in the study of intracellular signaling molecules and stimulate new research on antiviral mechanisms to
 develop effective strategies for plant protection against viruses.