Transgenic plants of tall fescue (Festuca arundinacea Schreb.) obtained by direct gene transfer to protoplasts

Biotechnology (N Y). 1992 Jun;10(6):691-6. doi: 10.1038/nbt0692-691.

Abstract

Chimeric hygromycin phosphotransferase (hph) and phosphinothricin acetyltransferase (bar) genes were introduced, using polyethylene glycol treatment, into protoplasts isolated from embryogenic cell suspension cultures of tall fescue (Festuca arundinacea Schreb.), a graminaceous plant that is an important forage crop in temperate pastures. Colonies resistant to either 200 mg/l hygromycin or 100 mg/l phosphinothricin, respectively, were recovered upon selection using bead-type culture systems. Stable integration of the transgenes in the genomes of plants regenerated from resistant callus clones was shown by Southern hybridization analysis. In situ hybridization of a labeled transgene-probe to metaphase chromosomes is shown for one transgenic primary regenerant. Expression of the transgenes in mature plants was demonstrated by HPH enzyme assay or by phosphinothricin-herbicide spraying.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / genetics*
  • Aminobutyrates / pharmacology
  • Base Sequence
  • Cells, Cultured
  • Cinnamates*
  • Drug Resistance
  • Gene Expression
  • Hygromycin B / analogs & derivatives
  • Hygromycin B / pharmacology
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Phosphotransferases (Alcohol Group Acceptor)*
  • Phosphotransferases / genetics*
  • Plants / drug effects
  • Plants / genetics*
  • Polyethylene Glycols / pharmacology
  • Polymerase Chain Reaction
  • Protoplasts / drug effects
  • Transfection*

Substances

  • Aminobutyrates
  • Cinnamates
  • Polyethylene Glycols
  • Hygromycin B
  • hygromycin A
  • phosphinothricin
  • Acetyltransferases
  • phosphinothricin N-acetyltransferase
  • Phosphotransferases
  • Phosphotransferases (Alcohol Group Acceptor)
  • hygromycin-B kinase