Abstract
Biogenesis of chloroplasts is essential for plant growth and development. A number of homozygous mutants lacking a chloroplast protein exhibit an albino phenotype. In general, it is challenging to grow albino Arabidopsis plants on soil until they set seeds. Homozygous albino mutants are usually obtained as progenies of heterozygous parents. Here, we describe a method of recovering seeds from the seedling lethal Arabidopsis mutant ppi2-2, which lacks the atToc159 protein import receptor at the outer envelope membrane of chloroplast. Using plastic containers, we were able to grow homozygous ppi2-2 plants until these set seed. Although the germination rate of the harvested seeds was relatively low, it was still sufficient to allow us to further analyze the ppi2-2 progeny. Using ppi2-2 homozygous seeds, we were able to analyze the role of plastid protein import in the light-regulated induction of nuclear genes. We propose that this method be applied to other seedling lethal Arabidopsis mutants to obtain homozygous seeds, helping us further investigate the roles of plastid proteins in plant growth and development.
Highlights
Plastids such as chloroplasts in photosynthetic plant cells are believed to have evolved from a cyanobacterium-like ancestor (Dyall et al, 2004)
We describe a method for generating viable seeds from the seedling lethal Arabidopsis mutant ppi2-2, which lacks the major protein import receptor of plastids (Bauer et al, 2000; Kakizaki et al, 2009)
APPLICATIONS This report describes a method for obtaining viable seeds from the seedling lethal ppi2-2 mutant using Ziploc® containers
Summary
Plastids such as chloroplasts in photosynthetic plant cells are believed to have evolved from a cyanobacterium-like ancestor (Dyall et al, 2004). The key player involved in delivering nuclear-encoded proteins into plastids is the translocon at the outer envelope membrane of chloroplasts (TOC) and the translocon at the inner envelope membrane of chloroplasts (TIC) complex (Inaba and Schnell, 2008; Li and Chiu, 2010; Jarvis and Lopez-Juez, 2013). Molecular genetic analysis of identified components using Arabidopsis indicated that these were indispensable for plastid biogenesis (Jarvis et al, 1998; Bauer et al, 2000; Chou et al, 2003; Constan et al, 2004a,b; Ivanova et al, 2004; Kubis et al, 2004; Inaba et al, 2005; Kovacheva et al, 2005; Teng et al, 2006; Kikuchi et al, 2013)
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