Transplastomics in Arabidopsis: Progress Toward Developing an Efficient Method

Kerry Ann Lutz, Arun Azhagiri, Pal Maliga

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

Abstract

Protocols developed for plastome engineering in Nicotiana tabacum rely on biolistic delivery of the transforming DNA to chloroplasts in intact leaf tissue; integration of the foreign DNA into the plastid genome by homologous recombination via flanking plastid DNA (ptDNA) targeting regions; and gradual dilution of non-transformed ptDNA during cultivation in vitro. Plastid transformation in Arabidopsis was obtained by combining the tobacco leaf transformation protocol with Arabidopsis-specific tissue culture and plant regeneration protocols. Because the leaf cells in Arabidopsis are polyploid, this protocol yielded sterile plants. Meristematic cells in a shoot apex or cells of a developing embryo are diploid. Therefore, we developed a regulated embryogenic root culture system that will generate diploid tissue for plastid transformation. This embryogenic culture system is created by steroid-inducible expression of the BABY BOOM transcription factor. Plastid transformation in Arabidopsis will enable the probing of plastid gene function, and the characterization of posttranscriptional mechanisms of gene regulation and the regulatory interactions of plastid and nuclear genes.

Original languageAmerican English
Title of host publicationChloroplast Research in Arabidopsis
Subtitle of host publicationMethods and Protocols, Volume I
PublisherHumana Press Inc.
Pages133-147
Number of pages15
ISBN (Print)9781617792335
DOIs
StatePublished - 2011

Publication series

NameMethods in Molecular Biology
Volume774

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Keywords

  • Arabidopsis thaliana
  • BABY BOOM
  • Dexamethasone
  • Plant regeneration
  • Plastid transformation
  • Steroid-inducible gene expression
  • Tissue culture

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