High throughput selection of antibiotic-resistant transgenic Arabidopsis plants

Anal Biochem. 2017 May 15:525:44-45. doi: 10.1016/j.ab.2017.02.017. Epub 2017 Feb 27.

Abstract

Kanamycin resistance is the most frequently used antibiotic-resistance marker for Arabidopsis transformations, however, this method frequently causes escape of untransformed plants, particularly at the high seedling density during the selection. Here we developed a robust high-density selection method using top agar for Arabidopsis thaliana. Top agar effectively suppressed growth of untransformed wild-type plants on selection media at high density. Survival of the transformed plants during the selection were confirmed by production of green true leaves and expression of a firefly luciferase reporter gene. Top agar method allowed selection using a large amount of seeds in Arabidopsis transformation.

Keywords: Antibiotic selection; Arabidopsis; Transformation.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Agar / chemistry*
  • Agar / metabolism
  • Anti-Bacterial Agents / pharmacology*
  • Arabidopsis / drug effects
  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Genetic Engineering / methods*
  • High-Throughput Screening Assays
  • Kanamycin Resistance / genetics*
  • Luciferases / metabolism
  • Plant Leaves / drug effects
  • Plant Leaves / genetics
  • Plant Leaves / metabolism*
  • Plants, Genetically Modified / drug effects
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Seeds / drug effects
  • Seeds / genetics
  • Seeds / metabolism
  • Transformation, Genetic

Substances

  • Anti-Bacterial Agents
  • Agar
  • Luciferases