Genotyping-by-sequencing (GBS) was performed with two F1 mapping populations of H. bulbosum (FBB_1xPB1 and FBC_3xFBD_5) to calculate genetic maps of H. bulbosum.

For genotyping the F1 populations GBS libraries were prepared from genomic DNA by using the two restriction enzymes MspI and PstI High-Fidelity. In order to produce F1 progeny, flowering ears of a single plant of each accession as well as the clonal plants of PB1 were bagged as described earlier. This resulted in two F1 mapping populations of 190 (FBB1xPB1) and 272 (FBC_3xFBD_5) individuals, respectively. So each mapping population goes back to a single individual of the accessions FBB, FBC, FBD and PB1, respectively. The F1 populations as well as the parental genotypes were subjected to genotyping-by-sequencing (GBS).