Fig. 2.

Overview of mutant mapping strategy using exome capture and sequencing. Each M₁ plant grown from EMS-mutagenized seed was self-pollinated to produce single M₂ plants, which were exome-sequenced to catalog induced mutations in the protein-coding regions (Krasileva et al. 2017). M₃ rows derived from each M₂ plant were screened to identify mutant phenotypes of interest (depicted in yellow). Subsequent M₄ seeds bulk-harvested from the selected M₃ row segregating for the mutant phenotype (red dotted box) were used as a mapping population. Barcoded sequencing libraries with unique indices were constructed for each of the selected individual M₄ plants. Libraries were subjected to exome capture and sequenced in multiplexed reactions. Bioinformatics pipeline for sequencing reads processing, mapping, and genotype calling. *IWGSC CSS reference supplemented with a de novo assembly of unmapped reads from Kronos (Krasileva et al. 2017). (Color figure online)