Isolation of embryo-specific mutants in Arabidopsis: genetic and phenotypic analysis.

Abstract

To perform an effective genetic screen for embryo-specific mutants is a prerequisite for understanding the molecular mechanisms that control Arabidopsis embryo development. Mutagenesis based on either T-DNA or transposon insertion has been successfully used in identifying embryonic mutants. We present here a typical genetic screen for putative embryonic mutants based on distorted Mendelian segregation ratio (2:1) and reduced seed set. It is advisable to examine whether the mutation also affects gametophytic functions by performing reciprocal crosses between wild type and the mutant. We also provide detailed explanations on the whole-mount clearing method, a simple but effective method for phenotypic analysis of mutant embryos blocked in certain steps during the process necessary for embryo viability and development.