Queensland Alliance of Agriculture and Food Innovation, The University of Queensland, St Lucia, QLD, Australia.
ARC Centre of Excellence for Plant Success in Nature and Agriculture, The University of Queensland, St Lucia, QLD, Australia.
Physiol Plant. 2024 Jul-Aug;176(4):e14449. doi: 10.1111/ppl.14449.
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.
植物育种家利用化学、生物和物理诱变剂来利用诱变,创造新的性状变异。许多广泛使用的高粱基因型遗传基础狭窄,这阻碍了使用传统育种进行的改进。因此,增强高粱基因组的多样性仍然是高粱育种者的主要优先事项。为了加速高粱的遗传改良,使用乙基甲磺酸(EMS)作为诱变剂,建立了一个由 15 万株 BTx623 自交系高粱突变体植物的种子组成的广泛文库。高粱突变体文库被混为 1498 个池(每个池约 100 个头)。在每个池中,从种子的一部分提取 DNA,并使用基于液滴数字 PCR 的 FIND-IT 技术进行筛选。使用 FIND-IT 筛选的所有 43 个核苷酸取代都被鉴定出来,这表明有可能在几天内鉴定出高粱优良品系中的任何 EMS 衍生突变。这个多样化的文库代表了有史以来最大的高粱突变体集合,估计涵盖了高粱基因组中所有可能的 EMS 诱导突变点的 240%。使用 FIND-IT,可以大大提高特定所需 EMS 衍生突变的识别速度,这是对传统反向遗传技术的重大升级。此外,与基因组编辑相比,将有价值的变体轻松整合到优良商业品系中是一个简单得多且成本低得多的过程。文库中的基因组变异将直接用作商业高粱应用的育种资源,从而增强对气候变化的适应能力,并在边缘环境中提高产量潜力。