Genetics, Vol. 159, 1751-1763, December 2001, Copyright © 2001

Insertional Mutagenesis of Genes Required for Seed Development in Arabidopsis thaliana

John McElvera, Iris Tzafrirb, George Auxa, Rebecca Rogersb, Carl Ashbya, Kelsey Smithb, Carla Thomasa, Amy Schetterb, Qing Zhoua, Mary Ann Cushmanb, John Tossberga, Todd Nickleb, Joshua Z. Levina, Marcus Lawa, David Meinkeb, and David Pattona
a Syngenta, Research Triangle Park, North Carolina 27709
b Department of Botany, Oklahoma State University, Stillwater, Oklahoma 74078

Corresponding author: David Meinke, Department of Botany, 104 Life Sciences East, Oklahoma State University, Stillwater, OK 74078., meinke{at}okstate.edu (E-mail)

Communicating editor: C. S. GASSER

The purpose of this project was to identify large numbers of Arabidopsis genes with essential functions during seed development. More than 120,000 T-DNA insertion lines were generated following Agrobacterium-mediated transformation. Transgenic plants were screened for defective seeds and putative mutants were subjected to detailed analysis in subsequent generations. Plasmid rescue and TAIL-PCR were used to recover plant sequences flanking insertion sites in tagged mutants. More than 4200 mutants with a wide range of seed phenotypes were identified. Over 1700 of these mutants were analyzed in detail. The 350 tagged embryo-defective (emb) mutants identified to date represent a significant advance toward saturation mutagenesis of EMB genes in Arabidopsis. Plant sequences adjacent to T-DNA borders in mutants with confirmed insertion sites were used to map genome locations and establish tentative identities for 167 EMB genes with diverse biological functions. The frequency of duplicate mutant alleles recovered is consistent with a relatively small number of essential (EMB) genes with nonredundant functions during seed development. Other functions critical to seed development in Arabidopsis may be protected from deleterious mutations by extensive genome duplications.




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