Plant Molecular Biology Reporter

, Volume 18, Issue 2, pp 143–144

The use of the luciferase reporter system forin planta gene expression studies

  • Wessel Van Leeuwen
  • Marc J. M. Hagendoorn
  • Tom Ruttink
  • Remco Van Poecke
  • Linus H. W. Van Der Plas
  • Alexander R. Van Der Krol
Publish by Abstract

DOI: 10.1007/BF02824024

Cite this article as:
Van Leeuwen, W., Hagendoorn, M.J.M., Ruttink, T. et al. Plant Mol Biol Rep (2000) 18: 143. doi:10.1007/BF02824024

Abstract

The properties of the firefly luciferase (LUC) make it a very good nondestructive reporter to quantify and image transgene promoter activity in plants. The short half-life of the LUC mRNA and protein, and the very limited regeneration of the LUC protein after reacting with luciferin, enables monitoring of changes in gene activity with a high time resolution. However, the ease at which luciferase activity is measuredin planta, using a light sensitive camera system (2D-luminometer), contrasts sharply with the complications that arise from interpreting the results. A variegated pattern of luciferase activity, that is often observed inin planta measurements, might either be caused by differences in influx, availability of the substrates (luciferin, oxygen, ATP) or by local differences in reporter gene activity. Here we tested the possible contribution of differences in the availability of each substrate to the variegatedin planta luciferase activity, and we show whenin planta luciferase activity is measured under substrate equilibrium conditions and can be related to the promoter activity of the reporter gene. Furthermore, we demonstrate the effects of protein stability, apparent half-life of luciferase activity, regeneration of luciferase and pH on thein vivo andin vitro luciferase measurements. The combined results give the prerequisites for the correct utilisation of the luciferase reporter system, especially forin vivo gene expression studies in plant research.

Key words

Cauliflower Mosaic Virus 35S promoterluciferase reporter systemPetunia hybridareporter genetransgene expressionvariegation

Copyright information

© International Society for Plant Molecular Biology 2000

Authors and Affiliations

  • Wessel Van Leeuwen
    • 1
  • Marc J. M. Hagendoorn
    • 1
  • Tom Ruttink
    • 2
  • Remco Van Poecke
    • 3
  • Linus H. W. Van Der Plas
    • 1
  • Alexander R. Van Der Krol
    • 1
  1. 1.Laboratory of Plant PhysiologyWageningen UniversityWageningenThe Netherlands
  2. 2.Laboratory of Molecular BiologyWageningen UniversityWageningenThe Netherlands
  3. 3.Laboratory of EntomologyWageningen UniversityWageningenThe Netherlands