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Virus‐mediated efficient induction of epigenetic modifications of endogenous genes with phenotypic changes in plants
Author(s) -
Kanazawa Akira,
Inaba Junichi,
Shimura Hanako,
Otagaki Shungo,
Tsukahara Sayuri,
Matsuzawa Akihiko,
Kim Bo Min,
Goto Kazunori,
Masuta Chikara
Publication year - 2011
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2010.04401.x
Subject(s) - biology , rna silencing , gene silencing , epigenetics , dna methylation , genetics , gene , rna interference , bisulfite sequencing , promoter , rna , microbiology and biotechnology , gene expression
Summary Gene silencing through transcriptional repression can be induced by targeting double‐stranded RNA (dsRNA) to a gene promoter. It has been reported that a transgene was silenced by targeting dsRNA to the promoter, and the silenced state was inherited to the progeny plant even after removal of the silencing inducer from cells. In contrast, no plant has been produced that harbors silenced endogenous gene after removal of promoter‐targeting dsRNA. Here, we show that heritable gene silencing can be induced by targeting dsRNA to the endogenous gene promoters in petunia and tomato plants, using the Cucumber mosaic virus (CMV)‐based vector. We found that efficient silencing of endogenous genes depends on the function of the 2b protein encoded in the vector virus, which has the ability to facilitate epigenetic modifications through the transport of short interfering RNA to nucleus. Bisulfite sequencing analyses on the targeted promoter in the virus‐infected and its progeny plants revealed that cytosine methylation was found not only at CG or CNG but also at CNN sites. The observed inheritance of asymmetric DNA methylation is quite unique, suggesting that plants have a mechanism to maintain even asymmetric methylation. This CMV‐based gene silencing system provides a useful tool to artificially modify DNA methylation in plant genomes and elucidate the mechanism for epigenetic controls.

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