Open AccessCRISPR/Cas9‐mediated tetra‐allelic mutation of the ‘Green Revolution’ SEMIDWARF‐1 ( SD‐1 ) gene confers lodging resistance in tef ( Eragrostis tef )
Author(s) -
Beyene Getu,
Chauhan Raj Deepika,
Villmer Justin,
Husic Nada,
Wang Ning,
Gebre Endale,
Girma Dejene,
Chanyalew Solomon,
Assefa Kebebew,
Tabor Girma,
Gehan Malia,
McGrone Michael,
Yang Meizhu,
Lenderts Brian,
Schwartz Chris,
Gao Huirong,
GordonKamm William,
Taylor Nigel J.,
MacKenzie Donald J.
Publication year - 2022
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.13842
Subject(s) - biology , tilling , crispr , genetics , frameshift mutation , genome editing , gene , allele , cultivar , genetically modified rice , indel , crop , genetically modified crops , mutation , horticulture , transgene , agronomy , single nucleotide polymorphism , genotype
Summary 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 1 and T 2 generations revealed that the sd‐1 lines have reduced culm and internode lengths with no reduction in either panicle or peduncle lengths. These characteristics are comparable with rice sd‐1 plants. Measurements of lodging, in greenhouse‐grown plants, showed that sd‐1 lines have significantly higher resistance to lodging at the heading stage compared with the controls. This is the first demonstration of the feasibility of high frequency genetic transformation and CRISPR/Cas9‐mediated genome editing in this highly valuable but neglected crop. The findings reported here highlight the potential of genome editing for the improvement of lodging resistance and other important traits in tef.