Study : Identification of the causative mutation (using Whole Genome Sequencing) in Arabidopsis EMS mutants showing subcellular trafficking defects

Identification

• Name: Identification of the causative mutation (using Whole Genome Sequencing) in Arabidopsis EMS mutants showing subcellular trafficking defects
• Identifier: dXJuOkVWQS9zdHVkeS9QUkpOQTYxMjY4OA==
• Data link: Link to this study on EBI European Nucleotide Archive
• Description: Goal of the study was to identify mutants resistant to auxin inhibition of endocytosis. Overall design: M2 generation of 5-day-old EMS-mutagenized PIN1:PIN1-GFP seedlings (progenies of 1, 920 M1) were co-treated with 10 μM NAA and 25 μM BFA for 1 hour and screened under the fluorescent microscope for individuals with PIN1- GFP signal accumulated in BFA bodies, therefore insensitive to auxin effect on endocytosis. In M3 generation, the candidates were confirmed, obtained alleles were back crossed to the parental line 3 times and the homozygous lines were selected showing clear cellular phenotype. For next generation sequencing, genomic DNA of 3 homozygous backcrossed individuals and the parental PIN1:PIN1-GFP plant were submitted to Vienna Biocenter Core Facility for whole-genome sequencing. Interactive tool, artMAP (Javorka et al., 2019) was used to map EMS induced mutations.
• Data files: SRR11308374; SRR11308373; SRR11308372; SRR11308371