1. CRISPR/Cas9-mediated tetra-allelic mutation of the 'Green Revolution' SEMIDWARF-1 (SD-1) gene confers lodging resistance in tef (Eragrostis tef)
- Author
-
Getu Beyene, Raj Deepika Chauhan, Justin Villmer, Nada Husic, Ning Wang, Endale Gebre, Dejene Girma, Solomon Chanyalew, Kebebew Assefa, Girma Tabor, Malia Gehan, Michael McGrone, Meizhu Yang, Brian Lenderts, Chris Schwartz, Huirong Gao, William Gordon‐Kamm, Nigel J. Taylor, and Donald J. MacKenzie
- Subjects
Gene Editing ,Plant Breeding ,Mutation ,Plant Science ,CRISPR-Cas Systems ,Eragrostis ,Genes, Plant ,Plants, Genetically Modified ,Agronomy and Crop Science ,Alleles ,Biotechnology - Abstract
Tef is a staple food and a valuable cash crop for millions of people in Ethiopia. Lodging is a major limitation to tef production, and for decades, the development of lodging resistant varieties proved difficult with conventional breeding approaches. We used CRISPR/Cas9 to introduce knockout mutations in the tef orthologue of the rice SEMIDWARF-1 (SD-1) gene to confer semidwarfism and ultimately lodging resistance. High frequency recovery of transgenic and SD-1 edited tef lines was achieved in two tef cultivars by Agrobacterium-mediated delivery into young leaf explants of gene editing reagents along with transformation and regeneration enhancing morphogenic genes, BABY BOOM (BBM) and WUSCHEL2 (WUS2). All of the 23 lines analyzed by next-generation sequencing had at least two or more alleles of SD-1 mutated. Of these, 83% had tetra-allelic frameshift mutations in the SD-1 gene in primary tef regenerants, which were inherited in subsequent generations. Phenotypic data generated on T
- Published
- 2021