Arabidopsis Genes Essential for Seedling Viability: Isolation of Insertional Mutants and Molecular Cloning

• Published in: Genetics (Austin) Vol. 159; no. 4; pp. 1765 - 1778
• Main Authors: Budziszewski, Gregory J, Lewis, Sharon Potter, Glover, Lyn Wegrich, Reineke, Jennifer, Jones, Gary, Ziemnik, Lisa Schlater, Lonowski, Jennifer, Nyfeler, Beat, Aux, George, Zhou, Qing, McElver, John, Patton, David A, Martienssen, Robert, Grossniklaus, Ueli, Ma, Hong, Law, Marcus, Levin, Joshua Z
• Format: Journal Article
• Language: English
• Published: United States Genetics Soc America 01-12-2001; Genetics Society of America
• Subjects: Arabidopsis - genetics; Arabidopsis thaliana; Cell Survival; Chloroplasts - metabolism; Cloning; Cloning, Molecular; DNA Transposable Elements - genetics; Models, Genetic; Molecular biology; Multigene Family; Mutagenesis, Insertional; Mutation; Phenotype; Plasmids - metabolism; Polymerase Chain Reaction; Seeds; Seeds - genetics; Seeds - physiology
• ISSN: 0016-6731; 1943-2631; 1943-2631
• DOI: 10.1093/genetics/159.4.1765

We have undertaken a large-scale genetic screen to identify genes with a seedling-lethal mutant phenotype. From screening approximately 38,000 insertional mutant lines, we identified >500 seedling-lethal mutants, completed cosegregation analysis of the insertion and the lethal phenotype for >200 mutants, molecularly characterized 54 mutants, and provided a detailed description for 22 of them. Most of the seedling-lethal mutants seem to affect chloroplast function because they display altered pigmentation and affect genes encoding proteins predicted to have chloroplast localization. Although a high level of functional redundancy in Arabidopsis might be expected because 65% of genes are members of gene families, we found that 41% of the essential genes found in this study are members of Arabidopsis gene families. In addition, we isolated several interesting classes of mutants and genes. We found three mutants in the recently discovered nonmevalonate isoprenoid biosynthetic pathway and mutants disrupting genes similar to Tic40 and tatC, which are likely to be involved in chloroplast protein translocation. Finally, we directly compared T-DNA and Ac/Ds transposon mutagenesis methods in Arabidopsis on a genome scale. In each population, we found only about one-third of the insertion mutations cosegregated with a mutant phenotype.