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Plant and Cell Physiology Advance Access published online on February 21, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm028
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© The Author 2007. Published by Oxford University Press on behalf of the Japanese Society of Plant Physiologists. All rights reserved.

Epigenetic inactivation of chalcone synthase-A transgene transcription in petunia leads to a reversion of the post-transcriptional gene silencing phenotype

Akira Kanazawa1,*, Michael O'Dell2 and Roger P. Hellens3

1 Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
2 Department of Genetics, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom
3 Gene Discovery and Function Department, HortResearch, Private Bag 92 169, Auckland, New Zealand

* Corresponding author: E-mail, kanazawa{at}res.agr.hokudai.ac.jp; Fax, +81-11-706-4933.


  Abstract

Petunia plants that exhibit a white-flowering phenotype as a consequence of chalcone synthase transgene-induced silencing occasionally produce revertant branches that produce flowers with wild-type pigmentation. Transcription run-on assays confirmed that the production of white flowers is caused by post-transcriptional gene silencing (PTGS), and indicated that transgene transcription is repressed in the revertant plants, providing evidence that induction of PTGS depends on transcription rate. Transcriptional repression of the transgene was associated with cytosine methylation at CpG, CpNpG, and CpNpN sites, and the expression was restored by treatment with either 5-azacytidine or trichostatin A. These results demonstrate that epigenetic changes occurred in the PTGS line, and these changes interfere with the initiation of transgene transcription, leading to a reversion of the PTGS phenotype.

Keywords: bisulfite sequencing analysis - cauliflower mosaic virus 35S promoter - chalcone synthase - cytosine methylation - Petunia hybrida - post-transcriptional gene silencing


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