Open AccessArabidopsis Genes Essential for Seedling Viability: Isolation of Insertional Mutants and Molecular Cloning
Author(s) -
Gregory J. Budziszewski,
Sharon Potter Lewis,
Lyn Wegrich Glover,
Jennifer Reineke,
Gary W. Jones,
Lisa Schlater Ziemnik,
Jennifer Lonowski,
Beat Nyfeler,
George Aux,
Qing Zhou,
John McElver,
David A. Patton,
Robert A. Martienssen,
Ueli Grossniklaus,
Hong Mā,
Marcus Law,
Joshua Z. Levin
Publication year - 2001
Publication title -
genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.792
H-Index - 246
eISSN - 1943-2631
pISSN - 0016-6731
DOI - 10.1093/genetics/159.4.1765
Subject(s) - biology , mutant , arabidopsis , genetics , insertional mutagenesis , gene , mutagenesis , transposable element , phenotype , positional cloning
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.