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Free Content Graft transmission of induced and spontaneous post-transcriptional silencing of chitinase genes

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Sense and antisense tobacco chitinase (CHN) transgenes, Luciferase-CHN transcriptional fusions, and promoterless CHN cDNAs were introduced biolistically into CHN transformants of tobacco that never exhibit spontaneous gene silencing. All of the constructs tested induced systemic silencing of the resident CHN transgene and endogenes. Nuclear run-on transcription assays showed that local introduction of additional gene copies triggers systemic post-transcriptional gene silencing (PTGS). Together, this provides evidence that additional transgene copies need not be either highly transcribed or produce sense transcripts to evoke production of systemic PTGS signals. CHN PTGS was transmitted by top grafting, but not by reciprocal grafting of mature stems or the exchange of tissue plugs. Thus, the commonly encountered difficulties in achieving graft-transmission could reflect the method used. Silencing in sense but not antisense transformants was transmitted by grafting to a high-expressing sense CHN scion suggesting that the elaboration of mobile signals may not be an essential feature of antisense-mediated gene silencing.
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Keywords: RNA interference; antisense transformation; chitinase; graft transmission; post-transcriptional gene silencing; systemic signals

Document Type: Research Article

Affiliations: The Friedrich Miescher Institute for Biomedical Research, A branch of the Novartis Research Foundation, Maulbeerstrasse 66, CH-4058, Basel Switzerland

Publication date: December 1, 2001

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