A Semi-In Vivo Transcriptional Assay to Dissect Plant Defense Regulatory Modules

Fatimah Aljedaani, Naganand Rayapuram, Ikram Blilou*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

3 Scopus citations


Plants use different regulatory modules in response to changes in their surroundings. With the transcriptomic approaches governing all research areas, an integrative, fast, and sensitive approach toward validating genes of interest becomes a critical step prior to functional studies in planta. This chapter describes a detailed method for a quantitative analysis of transcriptional readouts of defense response genes using tobacco leaves as a transient system. The method uses Luciferase reporter assays to monitor activities of defense pathway promoters. Under normal conditions, the JASMONATE ZIM-DOMAIN (JAZ) proteins repress defense genes by preventing their expression. Here, we will provide a detailed protocol on the use of a dual-luciferase system to analyze activities of various defense response promoters simultaneously. We will use two well-characterized modules from the Jasmonic acid (JA) defense pathway; the JAZ3 repressor protein and the promoters of three of JA responsive genes, MYC2, 3 and 4. This assay revealed not only differences in promoter strength but also provided quantitative insights on the JAZ3 repression of MYCs in a quantitative manner.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages12
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • DLR
  • Fluorescence
  • JAZ3
  • Luciferase
  • Luminescence
  • MYCs
  • Promoter
  • Tobacco
  • Transcriptional regulation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


Dive into the research topics of 'A Semi-In Vivo Transcriptional Assay to Dissect Plant Defense Regulatory Modules'. Together they form a unique fingerprint.

Cite this