RESUMO
Plant breeders leverage mutagenesis using chemical, biological, and physical mutagens to create novel trait variations. Many widely used sorghum genotypes have a narrow genetic base, which hinders improvements using classical breeding. Enhancing the diversity of the sorghum genome thus remains a key priority for sorghum breeders. To accelerate the genetic enhancement of sorghum, an extensive library comprised of seeds from 150,000 individual mutant plants of the Sorghum bicolor inbred line BTx623 was established using ethyl methanesulphonate (EMS) as a mutagen. The sorghum mutant library was bulked into 1498 pools (~100 seed heads per pool). In each pool, DNA was extracted from a subset of the seed and screened using the FIND-IT technology based on droplet digital PCR. All 43 nucleotide substitutions that were screened using FIND-IT were identified, demonstrating the potential to identify any EMS-derived mutation in an elite line of sorghum within days. This diverse library represents the largest collection of sorghum mutants ever conceived, estimated to cover 240% of all possible EMS-induced mutation points within the Sorghum genome. Using FIND-IT, the speed at which a specific desired EMS-derived mutation can be identified is a major upgrade to conventional reverse genetic techniques. Additionally, the ease at which valuable variants can be integrated into elite commercial lines is a far simpler and less expensive process compared to genome editing. Genomic variations in the library will have direct utility as a breeding resource for commercial sorghum applications, allowing enhanced adaptation to climate change and enhanced yield potential in marginal environments.