Basic Techniques to Assess Seed Germination Responses to Abiotic Stress in Arabidopsis thaliana

Methods Mol Biol. 2016:1398:183-96. doi: 10.1007/978-1-4939-3356-3_15.

Abstract

The model organism Arabidopsis thaliana has been extensively used to unmask the molecular genetic signaling pathways controlling seed germination in plants. In Arabidopsis, the normal seed to seedling developmental transition involves testa rupture soon followed by endosperm rupture, radicle elongation, root hair formation, cotyledon expansion, and greening. Here we detail a number of basic procedures to assess Arabidopsis seed germination in response to different light (red and far-red pulses), temperature (seed thermoinhibition), and water potential (osmotic stress) environmental conditions. We also discuss the role of the endosperm and how its germination-repressive activity can be monitored genetically by means of a seed coat bedding assay. Finally we detail how to evaluate germination responses to changes in gibberellin (GA) and abscisic acid (ABA) levels by manipulating pharmacologically the germination medium.

Keywords: Abscisic acid; Endosperm rupture; Gibberellins; Osmotic stress; Seed coat bedding assay; Seed germination; Testa rupture; Thermoinhibition.

MeSH terms

  • Abscisic Acid / pharmacology*
  • Arabidopsis / drug effects
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / metabolism
  • Endosperm / drug effects
  • Endosperm / metabolism
  • Gene Expression Regulation, Plant / drug effects
  • Germination / drug effects
  • Gibberellins / pharmacology
  • Seeds / drug effects
  • Seeds / metabolism

Substances

  • Arabidopsis Proteins
  • Gibberellins
  • Abscisic Acid