Effective production of marker-free transgenic strawberry plants using inducible site-specific recombination and a bifunctional selectable marker gene

J.G. Schaart, F.A. Krens, K.T.B. Pelgrom, O. Mendes, G.J.A. Rouwendal

Research output: Contribution to journalArticleAcademicpeer-review

119 Citations (Scopus)

Abstract

Public concerns about the issue of the environmental safety of genetically modified plants have led to a demand for technologies allowing the production of transgenic plants without selectable (antibiotic resistance) markers. We describe the development of an effective transformation system for generating such marker-free transgenic plants, without the need for repeated transformation or sexual crossing. This system combines an inducible site-specific recombinase for the precise elimination of undesired, introduced DNA sequences with a bifunctional selectable marker gene used for the initial positive selection of transgenic tissue and subsequent negative selection for fully marker-free plants. The described system can be generally applied to existing transformation protocols, and was tested in strawberry using a model vector in which site-specific recombination leads to a functional combination of a cauliflower mosaic virus 35S promoter and a GUS encoding sequence, thereby enabling the histochemical monitoring of recombination events. Fully marker-free transgenic strawberry plants were obtained following two different selection/regeneration strategies.
Original languageEnglish
Pages (from-to)233-240
JournalPlant Biotechnology Journal
Volume2
Issue number3
DOIs
Publication statusPublished - 2004

Keywords

  • cytosine deaminase
  • glucocorticoid receptor
  • transformation vector
  • negative selection
  • bacterial gene
  • expression
  • agrobacterium
  • regeneration
  • system
  • frequency

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