Efficient, Antibiotic Marker-Free Transformation of a Dicot and a Monocot Crop with Glutamate 1-Semialdehyde Aminotransferase Selectable Marker Genes

Methods Mol Biol. 2016:1385:89-98. doi: 10.1007/978-1-4939-3289-4_6.

Abstract

Antibiotic-free, efficient in vitro selection in plant genetic engineering can improve risk perception and speed up pre-market scrutiny of genetically modified crops. We provide a protocol for genetic transformation of two important crops, durum wheat and alfalfa, using a bacterial and a plant-derived selectable marker gene encoding mutated, gabaculine-insensitive glutamate 1-semialdehyde aminotransferase (GSA) enzymes. These methods can potentially be applied, with minor adaptations, to many other monocot and dicot crop plants.

Keywords: Alfalfa; Durum wheat; GMO; Gabaculine; HemL; MsGSAgr; Non-antibiotic selection; Plant genetic engineering.

MeSH terms

  • Bacterial Proteins / genetics
  • Genetic Engineering / methods*
  • Genetic Markers
  • Intramolecular Transferases / genetics*
  • Medicago sativa / genetics*
  • Medicago sativa / metabolism
  • Plant Proteins / genetics
  • Plants, Genetically Modified
  • Transformation, Genetic*
  • Triticum / genetics*
  • Triticum / metabolism

Substances

  • Bacterial Proteins
  • Genetic Markers
  • Plant Proteins
  • Intramolecular Transferases
  • glutamate-1-semialdehyde 2,1-aminomutase