Development of a non-lethal selection system by using the aadA marker gene for efficient recovery of transgenic rice (Oryza sativa L.)

A. S. Oreifig, G. Kovács, B. Jenes, O. Toldi, E. Kiss, P. Scott

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The application of aminoglycoside-3″-adenyl-transferase (aadA) gene-mediated streptomycin resistance for non-lethal selection of transgenic rice resulted in plant regeneration frequencies under selection pressure as high as those in non-transformed controls without selection. Since streptomycin does not kill non-transgenic cells, and allows plant regeneration from them, a selection procedure was developed that made the visual identification of transgenic calli and regenerants possible. For callus-level selection, a vital pH indicator - Chlorophenol Red - was applied together with streptomycin, making use of the phenomenon that fast-growing cell lines lower the pH in the culture medium. Transgenic plants were selected according to their main distinctive features; their green colour (photomixotrophic assimilation), and more intense growth. At the same time, non-transgenic regenerants were bleached (heterotrophic assimilation), and growth was retarded in the presence of streptomycin and sucrose. The final efficiency of genetic transformation based on streptomycin resistance was found to be double that of transformations where the selective agent was L-phosphinothricin, and nearly three times more compared to transformations resulting in hygromycin-resistant regenerants. To the best of our knowledge, this is the first report on producing nuclear transformed rice plants by using a non-lethal selection strategy based on the chimaeric aadA gene.

Original languageEnglish
Pages (from-to)490-496
Number of pages7
JournalPlant Cell Reports
Volume22
Issue number7
DOIs
Publication statusPublished - Feb 2004

Fingerprint

aminoglycosides
streptomycin
transferases
Oryza sativa
genetically modified organisms
rice
genetic markers
assimilation (physiology)
callus
chlorophenols
glufosinate
genetic transformation
transgenic plants
genes
culture media
cell lines
sucrose
color
cells

Keywords

  • Aminoglycoside-3″-adenyltransferase
  • Genetic transformation
  • Non-lethal selection
  • Rice
  • Streptomycin

ASJC Scopus subject areas

  • Plant Science

Cite this

Development of a non-lethal selection system by using the aadA marker gene for efficient recovery of transgenic rice (Oryza sativa L.). / Oreifig, A. S.; Kovács, G.; Jenes, B.; Toldi, O.; Kiss, E.; Scott, P.

In: Plant Cell Reports, Vol. 22, No. 7, 02.2004, p. 490-496.

Research output: Contribution to journalArticle

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