Generation of a Stress-Inducible Luminescent Arabidopsis and Its Use in Genetic Screening for Stress-Responsive Gene Deregulation Mutants

Methods Mol Biol. 2017:1631:109-119. doi: 10.1007/978-1-4939-7136-7_6.

Abstract

In order to understand plant stress tolerance and its application, it is important to identify the signaling components involved in the stress-regulated gene expression. One initial step for this is generation of a stress-inducible luminescent Arabidopsis and its use in genetic mutant screening. Here, we describe how to generate a transgenic Arabidopsis line harboring a single copy of the STABILIZED1 (STA1) promoter-driven luciferase transgene (STA1p-LUC) as an example. STA1 is a pre-mRNA splicing factor Prp6p homolog and is induced by cold and heat stresses. In addition, generation of the STA1p-LUC mutant pool and a luminescence imaging-based screening for STA1p-LUC deregulated mutants are described.

Keywords: Arabidopsis; Cold stress; EMS mutagenesis; Genetic screening; Heat stress; Luminescence imaging; STA1; STABLIZED1; Thermal stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Arabidopsis* / genetics
  • Arabidopsis* / metabolism
  • Gene Expression Regulation, Plant / genetics*
  • Luciferases* / biosynthesis
  • Luciferases* / genetics
  • Mutation*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Plants, Genetically Modified* / genetics
  • Plants, Genetically Modified* / metabolism
  • Promoter Regions, Genetic / genetics
  • Stress, Physiological*

Substances

  • Arabidopsis Proteins
  • Nuclear Proteins
  • STA1 protein, Arabidopsis
  • Luciferases