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Posttranscriptional gene silencing of gn1 in tobacco triggers accumulation of truncated gn1-derived RNA species

Published online by Cambridge University Press:  06 September 2001

KATIA LITIÈRE
Affiliation:
Laboratorium voor Genetica, Departement Genetica, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium Present address: Wellcome/CRC Institute, Tennis Court Road, Cambridge CB2 1QR, United Kingdom.
GERBEN J. VAN ELDIK
Affiliation:
Laboratorium voor Genetica, Departement Genetica, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium Plant Genetic Systems N.V., J. Plateaustraat 22, B-9000, Gent, Belgium
JOHN J.M.R. JACOBS
Affiliation:
Plant Genetic Systems N.V., J. Plateaustraat 22, B-9000, Gent, Belgium
MARC VAN MONTAGU
Affiliation:
Laboratorium voor Genetica, Departement Genetica, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium
MARC CORNELISSEN
Affiliation:
Plant Genetic Systems N.V., J. Plateaustraat 22, B-9000, Gent, Belgium
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Abstract

Posttranscriptional silencing of basic β-1,3-glucanase genes in the tobacco line T17 is manifested by reduced transcript levels of the gn1 transgene and homologous, endogenous basic β-1,3-glucanase genes. An RNA ligation-mediated rapid amplification of cDNA ends (RLM-RACE) technique was used to compare the 3′ termini of gn1 RNAs present in expressing (hemizygous and young homozygous) and silenced (mature homozygous) T17 plants. Full-length, polyadenylated gn1 transcripts primarily accumulated in expressing plants, whereas in silenced T17 plants, mainly 3′-truncated, nonpolyadenylated gn1 RNAs were detected. The relative abundance of these 3′-truncated gn1 RNA species gradually increased during the establishment of silencing in homozygous T17 plants. Similar 3′-truncated, nonpolyadenylated gn1 RNA products were observed in an independent case of β-1,3-glucanase posttranscriptional gene silencing. This suggests that these 3′-truncated gn1 RNAs are a general feature of tobacco plants showing posttranscriptional silencing of the gn1 transgene.

Type
Research Article
Information
RNA , Volume 5 , Issue 10 , October 1999 , pp. 1364 - 1373
Copyright
1999 RNA Society

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