Genebank 2.0 - GBS

The great efforts spent in the maintenance of past diversity in genebanks are rationalized by the potential role of plant genetic resources (PGR) in future crop improvement. Molecular passport data derived from genomic sequence information can be used as a precise genebank management tool beyond the boundaries of classical descriptors - even across genebanks. Here, genotyping-by-sequencing (GBS) was carried out for 7,745 PGR samples that trace back to 7,651 winter wheat accessions from the German Federal ex situ genebank at IPK Gatersleben. Sequenced PGR plants were obtained by bagging one (two) representative ear(s) in case of homogeneous (clearly non-homogenous) PGR during seed multiplication in the field. In parallel, GBS was performed for 325 modern cultivars covering the diversity available at breeding programs across Europe. For 171 of these cultivars, three plants per genotype were independently sequenced, while a single plant per genotype was used for the remaining ones. DNA samples were digested with PstI and MspI. Barcoded DNA samples were pooled into batches of 540 genotypes in equimolar amount to form a GBS library. Single-end sequencing (1 × 107 cycles) was conducted following Illumina protocols on HiSeq 2000 (171 × 3 samples), HiSeq 2500 (6,262 samples) or NovaSeq 6000 (1,637 samples) devices and using custom sequencing primers. Passport data for these samples were collected from different sources by Schulthess et al. 2022.