Submitted to: Plant and Animal Genome Conference
Publication Type: Abstract Only
Publication Acceptance Date: 11/8/2019
Publication Date: 1/11/2020
Citation: Xin, Z., Burow, G.B., Hayes, C.M., Emendack, Y., Chen, J., Ware, D. 2020. Pedigreed mutant library as an efficient resource to discover targets for genome editing. Plant and Animal Genome Conference. PE0768.
Technical Abstract: Mutagenesis has been a classical and effective approach to dissect metabolic, developmental, and signal transduction pathways in both plants and animals due to the high density and random nature of induced mutations. A manageable number of lines can yield large number of mutations covering the entire genome. However, it suffers from two main disadvantages: 1) the lengthy and expensive process to identify the causal mutation and 2) the high density of background mutations. These two disadvantages have been largely overcome by the development in Next-Generation-Sequencing (NGS) and precise and efficient genome-editing technologies. We have established Pedigreed Mutant Library in the sorghum inbred line BTx623 by mutagenizing the seeds with ethyl methane sulfonate (EMS). This library has 6,400 M4 seed pools and possesses a great diversity of mutant phenotypes. A large number of sorghum mutants with altered agronomic traits has been isolated from the mutant library. We have established an effective bioinformatic pipeline to identify the causal mutations through bulk-segregant-analysis (BSA) of the whole genome sequencing data of the pooled mutants selected from F2 populations. Once an F2 backcrossed population is established, the cost to identify the causal mutation is under $300 in NGS sequencing. In the last few years, we have identified over 30 causal mutations underlying the altered agronomic traits. These causal mutations can serve as targets for genome-editing in elite sorghum lines. The combination of high throughput gene discovery from mutant library with precise genome editing will truly revolutionize plant and animal breeding.