Genetic mapping of the pendant awn trait using Genotyping-by-Sequencing of F2 individuals segregating for the mutant phenotype

Genotyping-by-sequencing (GBS) was performed on F2 individuals segregating for the pendant awn mutant trait, along with wild-type controls (Alraune, Igri) and the mutant line M4IGRI_11. Genomic DNA was digested with PstI-HF and MspI, size-selected (150–600 bp), quality-checked, and sequenced on an Illumina NovaSeq6000 platform (single-end, 138 cycles, 2x8 index cycles) to generate SNP markers for genetic analysis.