Low frequency of T-DNA based activation tagging in Arabidopsis is correlated with methylation of CaMV 35S enhancer sequences

A. Chalfun-Junior, J.J. Mes, L. Mlynarova, M.G.M. Aarts, G.C. Angenent

Research output: Contribution to journalArticleAcademicpeer-review

23 Citations (Scopus)

Abstract

A powerful system to create gain-of-function mutants in plants is activation tagging using T-DNA based vehicles to introduce transcriptional enhancer sequences. Large Arabidopsis populations of individual plants carrying a quadruple cauliflower mosaic virus (CaMV) 35S enhancer are frequently used for mutant screenings, however the frequency of morphological mutants remains very low. To clarify this low frequency we analyzed a subset of lines generated by this method. The correlation between the number of T-DNA insertion sites, the methylation status of the 35S enhancer sequence and 35S enhancer activity was determined. All plants containing more than a single T-DNA insertion showed methylation of the 35S enhancer and revealed a dramatic decrease in 35S enhancer activity. The results support the notion that in a large proportion of the T-DNA based activation tagged lines the 35S transcriptional enhancer is silenced due to methylation, which is induced by multiple T-DNA integrations.
Original languageEnglish
Pages (from-to)459-463
JournalFEBS Letters
Volume555
Issue number3
DOIs
Publication statusPublished - 2003

Keywords

  • double-stranded-rna
  • gene
  • plants
  • inheritance
  • thaliana
  • transgenes
  • growth

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