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Fate of hairpin transcript components during RNA silencing and its suppression in transgenic virus-resistant tobacco

Paper ID Volume ID Publish Year Pages File Format Full-Text
25660 43589 2006 8 PDF Available
Title
Fate of hairpin transcript components during RNA silencing and its suppression in transgenic virus-resistant tobacco
Abstract

Transgenic tobacco plants, carrying a Potato virus Y (PVY)-NIa hairpin sequence separated by a unique unrelated spacer sequence were specifically silenced and highly resistant to PVY infection. In such plants neither PVY-NIa nor spacer transgene transcripts were detectable by specific quantitative real time reverse transcriptase PCR (RT-qPCR) assays of similar relative efficiencies developed for direct comparative analysis. However, small interfering RNAs (siRNAs) specific for the PVY sequence of the transgene and none specific for the LNYV spacer sequence were detected. Following infection with Cucumber mosaic virus (CMV), which suppresses dsRNA-induced RNA silencing, transcript levels of PVY-NIa as well as spacer sequence increased manifold with the same time course. The cellular abundance of the single-stranded (ss) spacer sequence was consistently higher than that of PVY dsRNA in all cases. The results show that during RNA silencing and its suppression of a hairpin transcript in transgenic tobacco, the ssRNA spacer sequence is affected differently than the dsRNA. In PVY-silenced plants, the spacer is efficiently degraded by a mechanism not involving the accumulation of siRNAs, while following suppression of RNA silencing by CMV, the spacer appears protected from degradation.

Keywords
Post-transcriptional gene silencing; Real time RT-PCR; RNA silencing suppressor; Double stranded RNA
First Page Preview
Fate of hairpin transcript components during RNA silencing and its suppression in transgenic virus-resistant tobacco
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 2, 1 November 2006, Pages 115–122
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering