Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant

Delivery into plants of T-DNAs containing promoter, terminator, or coding sequences generated small interfering RNAs (siRNAs) specific to each type of sequence. When both promoter and transcribed sequences were simultaneously present in the T-DNA, accumulation of siRNAs to transcribed sequences was...

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Main Authors: Tomas Canto, Fabrizio Cillo, Peter Palukaitis
Format: Article
Language:English
Published: The American Phytopathological Society 2002-11-01
Series:Molecular Plant-Microbe Interactions
Online Access:https://apsjournals.apsnet.org/doi/10.1094/MPMI.2002.15.11.1137
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author Tomas Canto
Fabrizio Cillo
Peter Palukaitis
author_facet Tomas Canto
Fabrizio Cillo
Peter Palukaitis
author_sort Tomas Canto
collection DOAJ
description Delivery into plants of T-DNAs containing promoter, terminator, or coding sequences generated small interfering RNAs (siRNAs) specific to each type of sequence. When both promoter and transcribed sequences were simultaneously present in the T-DNA, accumulation of siRNAs to transcribed sequences was favored over accumulation of siRNAs to the nontranscribed upstream promoter sequences. The generation of specific siRNA sequences occurred even in the absence of T-DNA homology to sequences in the plant. Delivery of T-DNA, with homology to the transgene limited to the nontranscribed cauliflower mosaic virus 35S promoter (35SP) and the transcribed nopaline synthase transcription termination (NosT)signal sequences, into transgenic plants expressing the green fluorescent protein (GFP), generated siRNAs in infiltrated tissues to both 35SP (35SsiRNAs) and NosT (NosTsiRNAs), but not to the GFP sequence (GFPsiRNAs). In infiltrated tissues, the 35SsiRNAs failed to trigger the transcriptional silencing of the transgene, accumulation of 35SsiRNAs could be prevented by the potyviral HC-Pro, and the NosTsiRNAs required an initial amplification to trigger efficient transgene silencing, which is mediated by transcripts from the exogenous T-DNA, and not from the transgene. In upper leaves, silencing correlated with the presence of GFPsiRNAs and the absence of 35SsiRNAs, confirming that its spread was posttranscriptionally mediated by the transgene mRNA.
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spelling doaj.art-bd0d957133bf46ef940dfe50a3fc54632022-12-22T03:02:31ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77062002-11-0115111137114610.1094/MPMI.2002.15.11.1137Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the PlantTomas CantoFabrizio CilloPeter PalukaitisDelivery into plants of T-DNAs containing promoter, terminator, or coding sequences generated small interfering RNAs (siRNAs) specific to each type of sequence. When both promoter and transcribed sequences were simultaneously present in the T-DNA, accumulation of siRNAs to transcribed sequences was favored over accumulation of siRNAs to the nontranscribed upstream promoter sequences. The generation of specific siRNA sequences occurred even in the absence of T-DNA homology to sequences in the plant. Delivery of T-DNA, with homology to the transgene limited to the nontranscribed cauliflower mosaic virus 35S promoter (35SP) and the transcribed nopaline synthase transcription termination (NosT)signal sequences, into transgenic plants expressing the green fluorescent protein (GFP), generated siRNAs in infiltrated tissues to both 35SP (35SsiRNAs) and NosT (NosTsiRNAs), but not to the GFP sequence (GFPsiRNAs). In infiltrated tissues, the 35SsiRNAs failed to trigger the transcriptional silencing of the transgene, accumulation of 35SsiRNAs could be prevented by the potyviral HC-Pro, and the NosTsiRNAs required an initial amplification to trigger efficient transgene silencing, which is mediated by transcripts from the exogenous T-DNA, and not from the transgene. In upper leaves, silencing correlated with the presence of GFPsiRNAs and the absence of 35SsiRNAs, confirming that its spread was posttranscriptionally mediated by the transgene mRNA.https://apsjournals.apsnet.org/doi/10.1094/MPMI.2002.15.11.1137
spellingShingle Tomas Canto
Fabrizio Cillo
Peter Palukaitis
Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant
Molecular Plant-Microbe Interactions
title Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant
title_full Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant
title_fullStr Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant
title_full_unstemmed Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant
title_short Generation of siRNAs by T-DNA Sequences Does Not Require Active Transcription or Homology to Sequences in the Plant
title_sort generation of sirnas by t dna sequences does not require active transcription or homology to sequences in the plant
url https://apsjournals.apsnet.org/doi/10.1094/MPMI.2002.15.11.1137
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AT fabriziocillo generationofsirnasbytdnasequencesdoesnotrequireactivetranscriptionorhomologytosequencesintheplant
AT peterpalukaitis generationofsirnasbytdnasequencesdoesnotrequireactivetranscriptionorhomologytosequencesintheplant