Protocol

EMS Mutagenesis of Arabidopsis Seed

This protocol was adapted from "Obtaining Mutants," Chapter 2 in Arabidopsis by Detlef Weigel and Jane Glazebrook. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2002.

INTRODUCTION

EMS-mutagenized Arabidopsis seed collections can be purchased from companies or acquired from other researchers. But in some cases (e.g., mutagenesis of a specific genotype), it is necessary for an investigator to generate EMS-mutagenized seed. In this protocol, EMS is used at concentrations that induce multiple point mutations in each plant, such that mutant alleles of a specific locus are found at a rate of ~1 in 2000-5000 M₂ plants. This high rate of mutagenesis makes possible the screening of relatively few plants to find those with the phenotype of interest, a particular advantage if the screen is laborious or if only a small number of genes mutate to the required phenotype.