Insertional Mutagenesis of Genes Required for Seed Development in Arabidopsis thaliana

Abstract

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.

E3 ligase BRUTUS Is a Negative Regulator for the Cellular Energy Level and the Expression of Energy Metabolism-Related Genes Encoded by Two Organellar Genomes in Leaf Tissues

“…A previous study showed that the knock-out mutation of BTS is lethal ( McElver et al, 2001 ; Selote et al, 2015 ; Tzafrir et al, 2004 ). This result raises the possibility that BTS plays a role in various developmental stages.…”

The pentatricopeptide repeat protein MTSF3 is required fornad2mRNA stability and embryogenesis in Arabidopsis

“…Arabidopsis ( Arabidopsis thaliana ) Col-0 plants were obtained from the Arabidopsis stock centre of the Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement in Versailles (http://dbsgap.versailles.inra.fr/portail/). T-DNA insertion line mstf3 (N816708) mutants was obtained from the European Arabidopsis Stock Center (NASC) (McElver et al, 2001). The mtsf3 mutant plants were genotyped by PCR using the primers listed in Supporting Information Table S1 and the insertion site was confirmed by sequencing.…”

Analytical Method Evaluation and Discovery of Variation within Maize Varieties in the Context of Food Safety: Transcript Profiling and Metabolomics

“…Three types of tissues, V5 leaf, 25 DAP immature kernel, and mature kernel, were used for metabolomic experiments. Because the mature kernels are dormant and have very limited gene expression, , only the V5 leaf and the 25 DAP immature kernels were used for microarray experiments. Due to limited tissue availability for some varieties, some microarray or metabolomic experiments were not conducted.…”