Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana

Abstract

Over 225,000 independent Agrobacterium transferred DNA (T-DNA) insertion events in the genome of the reference plant Arabidopsis thaliana have been created that represent near saturation of the gene space. The precise locations were determined for more than 88,000 T-DNA insertions, which resulted in the identification of mutations in more than 21,700 of the ∼29,454 predicted Arabidopsis genes. Genome-wide analysis of the distribution of integration events revealed the existence of a large integration site bias at both the chromosome and gene levels. Insertion mutations were identified in genes that are regulated in response to the plant hormone ethylene.

A Genetic Screen for Leaf Movement Mutants Identifies a Potential Role for AGAMOUS-LIKE 6 (AGL6) in Circadian-Clock Control

“…1F and 1G). These results were consistent with previously reported data (Alonso et al, 2003;Mizoguchi et al, 2002;Somers et al, 1998), suggesting that our imaging system is in fact an accurate method for screening leaf movement mutants.…”

“…In addition, phototropins (PHOTs), ZEITLUPE (ZTL), and LOV (LIGHT OXYGEN OR VOLTAGE)/KELCH PROTEIN 2 (LKP2) may also actively mediate additional light input. ZTL is a circadian photoreceptor stabilized by GI in blue light; it has the ability to degrade TOC1 protein (Alonso et al, 2003;Kim et al, 2007). The circadian clock can be also entrained by temperature cycles according to the temperature difference between daytime and nighttime (Alonso et al, 2003;Devlin, 2002;Millar, 2004).…”

“…ZTL is a circadian photoreceptor stabilized by GI in blue light; it has the ability to degrade TOC1 protein (Alonso et al, 2003;Kim et al, 2007). The circadian clock can be also entrained by temperature cycles according to the temperature difference between daytime and nighttime (Alonso et al, 2003;Devlin, 2002;Millar, 2004). Genes in the output pathway are the clock-regulated genes whose function is not necessary for endogenous circadian oscillation.…”

The inositol oxygenase gene family of Arabidopsis is involved in the biosynthesis of nucleotide sugar precursors for cell-wall matrix polysaccharides

“…For some of the MIOX genes, T-DNA insertion lines (DMIOX1, DMIOX2 and DMIOX5) were isolated from the SALK collection (Alonso et al 2003) or the GABI-KAT collection (Rosso et al 2003, http://www.gabikat.de). Homozygous knock-outs in single isoforms were analyzed for phenotypic changes compared to the wild type.…”

T-DNA–mediated transfer of Agrobacterium tumefaciens chromosomal DNA into plants

“…Our discovery that this sequence is identical to a region on the sequenced linear chromosome of A. tumefaciens led us to determine whether this was a unique event or whether it is an intrinsic property associated with T-DNA transfer in general. Therefore, we analyzed databases that contain A. thaliana-flanking sequence tags (FSTs) [6][7][8][9] , the sequences that flank T-DNA insertion sites in populations of insertion lines generated to saturate the genome with mutations. Fragments of AchrDNAs were detected in all tested T-DNA insertion databases, and AchrDNAs were found much more frequently in FSTs recovered from the RB ( Table 1).…”