Identifying a Growth Restoring Mutant in ala4/5 Dwarf Plants with PCR Diagnostics

Abstract

Aminophospholipid ATPases (ALAs) are lipid flippases that are involved in the uptake and translocation of specific lipids across membrane bilayers. Research demonstrates that ALA4 and ALA5 are critical to proper vegetative tissue development in Arabidopsis Thaliana. If an Arabidopsis Thaliana plant were to undergo a mutation removing ALA4 and ALA5 this plant would demonstrate vegetative dwarfism. Using a chemical mutagen in ala4/5 dwarf mutants four independent lines with restored growth were isolated and named suppressed ala4/5 dwarf mutants (SADs). The mutation for SAD3 was determined to have 43k candidates, which had to be significantly lowered to find the SAD3 mutation. Using PCR-diagnostic driven breeding approaches that exploit mutations that disrupt restriction digest sites can assist in the removal of non-causal candidate mutations in SAD3 mutants. Using this experimental model to contribute to the identification of the causal SAD3 mutation, plant lines were identified that lacked the flanking candidates.