Selective methylation of one of the two promoters residing in T-DNA inserted in a retroelement of rice

Selective methylation of one of the two promoters residing in T-DNA inserted in a retroelement of... The upstream sequence of pinb previously isolated from rice and confirmed to be a wound-inducible promoter by detecting GUS in T0 transgenic rice transformed via Agrobacterium tumefaciens-mediated procedures. In a transgenic line (pinb-16), the selectable marker hptII driven by CaMV35S promoter was completely silenced in T2 sublines; but the uidA gene driven by pinb promoter was expressed without being affected, though it, together with hptII, exists in the same T-DNA insertion. Analyses of methylation patterns using bisulphite-sequencing in the homozygous T1 and T2 sublines showed that cytosines in CaMV35S were gradually methylated in T1 plants and almost completely methylated in T2 plants. Interestingly, the process of methylation was accompanied by the occurrence of lesion mimic phenotype in rice leaves. The activity of hygromycin-resistance could be reestablished by treatment with 5-azacytidine. Genomic Southern and isolation of the T-DNA flanking sequences indicated that T-DNA was inserted in a retroelement of rice. These results revealed that methylation shows preference for the heterogeneity promoter fragment in the transgenic rice line and may be induced by the retroelement. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

Selective methylation of one of the two promoters residing in T-DNA inserted in a retroelement of rice

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Publisher
Springer Journals
Copyright
Copyright © 2006 by MAIK “Nauka/Interperiodica”
Subject
Life Sciences; Plant Sciences; Plant Physiology
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1134/S1021443706020142
Publisher site
See Article on Publisher Site

Abstract

The upstream sequence of pinb previously isolated from rice and confirmed to be a wound-inducible promoter by detecting GUS in T0 transgenic rice transformed via Agrobacterium tumefaciens-mediated procedures. In a transgenic line (pinb-16), the selectable marker hptII driven by CaMV35S promoter was completely silenced in T2 sublines; but the uidA gene driven by pinb promoter was expressed without being affected, though it, together with hptII, exists in the same T-DNA insertion. Analyses of methylation patterns using bisulphite-sequencing in the homozygous T1 and T2 sublines showed that cytosines in CaMV35S were gradually methylated in T1 plants and almost completely methylated in T2 plants. Interestingly, the process of methylation was accompanied by the occurrence of lesion mimic phenotype in rice leaves. The activity of hygromycin-resistance could be reestablished by treatment with 5-azacytidine. Genomic Southern and isolation of the T-DNA flanking sequences indicated that T-DNA was inserted in a retroelement of rice. These results revealed that methylation shows preference for the heterogeneity promoter fragment in the transgenic rice line and may be induced by the retroelement.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Mar 24, 2006

References

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