CRISPR/Cas9‐mediated tetra‐allelic mutation of the ‘Green Revolution’ SEMIDWARF‐1 (SD‐1) gene confers lodging resistance in tef (Eragrostis tef)

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...

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Published in:Plant biotechnology journal Vol. 20; no. 9; pp. 1716 - 1729
Main Authors: 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, Gordon‐Kamm, William, Taylor, Nigel J., MacKenzie, Donald J.
Format: Journal Article
Language:English
Published: England John Wiley & Sons, Inc 01.09.2022
John Wiley and Sons Inc
Subjects:
Agricultural production
Alleles
Amino acids
BABY BOOM
Biosynthesis
biotechnology
Cash crops
Cereals
Corn
CRISPR
CRISPR-Cas Systems
CRISPR/Cas9
Crops
culms
Cultivars
Domestication
Editing
Eragrostis - genetics
Eragrostis tef
Ethiopia
Explants
Frameshift mutation
Gene Editing
Genes
Genes, Plant
Genetic modification
Genetic transformation
genetically modified organisms
Genome editing
Genomes
Germplasm
Grain
Green revolution
High frequencies
Kinases
leaves
Lodging
Mutagenesis
Mutation
panicles
peduncle
phenotype
Plant Breeding
Plants, Genetically Modified - genetics
Proteins
Reagents
Rice
SEMIDWARF‐1
Sorghum
staple foods
WUSCHEL
Ethiopia
Eragrostis tef
lodging
BABY BOOM
WUSCHEL
CRISPR/Cas9
SEMIDWARF-1
ISSN:1467-7644, 1467-7652, 1467-7652
Online Access:Get full text
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Abstract 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 T1 and T2 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.
AbstractList 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 T1 and T2 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.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 T1 and T2 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.
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 and T 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.
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 T1 and T2 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.
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 T1 and T2 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.
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.
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 T₁ and T₂ 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.
Author Assefa, Kebebew
Chauhan, Raj Deepika
Lenderts, Brian
Husic, Nada
Wang, Ning
Yang, Meizhu
MacKenzie, Donald J.
Villmer, Justin
Gebre, Endale
Girma, Dejene
Tabor, Girma
Chanyalew, Solomon
Beyene, Getu
Gehan, Malia
Gordon‐Kamm, William
Gao, Huirong
Taylor, Nigel J.
McGrone, Michael
Schwartz, Chris
AuthorAffiliation 4 128161 Ethiopian Institute of Agricultural Research Addis Ababa Ethiopia
1 7538 Donald Danforth Plant Science Center St. Louis MO USA
2 57705 Corteva Agriscience Johnston IA USA
3 3078 Michigan State University East Lansing MI USA
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/35560779$$D View this record in MEDLINE/PubMed
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ContentType Journal Article
Copyright 2022 The Authors. published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.
2022 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.
2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
Copyright_xml – notice: 2022 The Authors. published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.
– notice: 2022 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.
– notice: 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
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DocumentTitleAlternate Green Revolution tef (Eragrostis tef)
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Issue 9
Keywords Eragrostis tef
lodging
BABY BOOM
WUSCHEL
CRISPR/Cas9
SEMIDWARF-1
Language English
License Attribution
2022 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.
This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
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Snippet 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...
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...
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proquest
pubmed
crossref
wiley
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Index Database
Enrichment Source
Publisher
StartPage 1716
SubjectTerms Agricultural production
Alleles
Amino acids
BABY BOOM
Biosynthesis
biotechnology
Cash crops
Cereals
Corn
CRISPR
CRISPR-Cas Systems
CRISPR/Cas9
Crops
culms
Cultivars
Domestication
Editing
Eragrostis - genetics
Eragrostis tef
Ethiopia
Explants
Frameshift mutation
Gene Editing
Genes
Genes, Plant
Genetic modification
Genetic transformation
genetically modified organisms
Genome editing
Genomes
Germplasm
Grain
Green revolution
High frequencies
Kinases
leaves
Lodging
Mutagenesis
Mutation
panicles
peduncle
phenotype
Plant Breeding
Plants, Genetically Modified - genetics
Proteins
Reagents
Rice
SEMIDWARF‐1
Sorghum
staple foods
WUSCHEL
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Title CRISPR/Cas9‐mediated tetra‐allelic mutation of the ‘Green Revolution’ SEMIDWARF‐1 (SD‐1) gene confers lodging resistance in tef (Eragrostis tef)
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Volume 20
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