Fluorescent antibiotic resistance marker for tracking plastid transformation in higher plants

Muhammad Sarwar Khan, Pal Maliga

Research output: Contribution to journalArticlepeer-review

176 Scopus citations

Abstract

Plastid transformation in higher plants is accomplished through a gradual process, during which all the 300-10,000 plastid genome copies are uniformly altered. Antibiotic resistance genes incorporated in the plastid genome facilitate maintenance of transplastomes during this process. Given the high number of plastid genome copies in a cell, transformation unavoidably yields chimeric tissues, which requires the identification of transplastomic cells in order to regenerate plants. In the chimeric tissue, however, antibiotic resistance is not cell autonomous: transplastomic and wild-type sectors both have a resistant phenotype because of phenotypic masking by the transgenic cells. We report a system of marker genes for plastid transformation, termed FLARE-S, which is obtained by translationally fusing aminoglycoside 3''-adenyltransferase with the Aequorea victoria green fluorescent protein. 3''-adenyltransferase (FLARE-S) confers resistance to both spectinomycin and streptomycin. The utility of FLARE-S is shown by tracking segregation of individual transformed and wild-type plastids in tobacco and rice plants after bombardment with FLARE-S vector DNA and selection for spectinomycin and streptomycin resistance, respectively. This method facilitates the extension of plastid transformation to nongreen plastids in embryogenic cells of cereal crops.

Original languageEnglish (US)
Pages (from-to)910-915
Number of pages6
JournalNature biotechnology
Volume17
Issue number9
DOIs
StatePublished - Sep 1999

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Keywords

  • Aminoglycoside 3''-adenylyl transferase
  • Green fluorescent protein
  • Plastid transformation
  • Rice
  • Spectinomycin and streptomycin resistance
  • Tobacco

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