WGS shallow sequencing of flowering time locus mutant F2 mapping populations

Flowering time locus (FLC) mutant F2 mapping populations were generated as follows. KO mutant lines covering most of the flowering time range in the mutant population were crossed to generate F1 progeny. Per cross, around five F1 plants were selfed to obtain biparental F2 mapping populations, from which one was randomly selected for the mapping experiment. All plants were propagated at 22°C under long days (LD, 16 h light / 8 h dark). The identity of the parents of the mutant lines and of the F1 individuals was verified with shallow Illumina short-read whole-genome sequencing. F2 plants were subsequently cultivated in trays containing 60 pots, with four trays assigned to each F2 mapping population. Six plants from each parent line were randomly placed across the four trays. While all trays for a mapping population were kept together in the greenhouse, they underwent regular rotation, and the entire group of trays was relocated every second day to minimize positional effects. Due to insufficient germination in certain pots, the F2 plant count per population varied.