Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties
CRISPR/Cas technology has recently become the molecular tool of choice for gene function studies in plants as well as crop improvement. Wheat is a globally important staple crop with a well annotated genome and there is plenty of scope for improving its agriculturally important traits using genome e...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-07-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/10/7/1481 |
| _version_ | 1797526322542542848 |
|---|---|
| author | Florian Hahn Laura Sanjurjo Loures Caroline A. Sparks Kostya Kanyuka Vladimir Nekrasov |
| author_facet | Florian Hahn Laura Sanjurjo Loures Caroline A. Sparks Kostya Kanyuka Vladimir Nekrasov |
| author_sort | Florian Hahn |
| collection | DOAJ |
| description | CRISPR/Cas technology has recently become the molecular tool of choice for gene function studies in plants as well as crop improvement. Wheat is a globally important staple crop with a well annotated genome and there is plenty of scope for improving its agriculturally important traits using genome editing technologies, such as CRISPR/Cas. As part of this study we targeted three different genes in hexaploid wheat <i>Triticum aestivum</i>: <i>TaBAK1-2</i> in the spring cultivar Cadenza as well as <i>Ta-eIF4E</i> and <i>Ta-eIF(iso)4E</i> in winter cultivars Cezanne, Goncourt and Prevert. Primary transgenic lines carrying CRISPR/Cas-induced indels were successfully generated for all targeted genes. While BAK1 is an important regulator of plant immunity and development, Ta-eIF4E and Ta-eIF(iso)4E act as susceptibility (S) factors required for plant viruses from the <i>Potyviridae</i> family to complete their life cycle. We anticipate the resultant homozygous <i>tabak1-2</i> mutant lines will facilitate studies on the involvement of BAK1 in immune responses in wheat, while <i>ta-eif4e</i> and <i>ta-eif(iso)4e</i> mutant lines have the potential to become a source of resistance to wheat spindle streak mosaic virus (WSSMV) and wheat yellow mosaic virus (WYMV), both of which are important pathogens of wheat. As winter wheat varieties are generally less amenable to genetic transformation, the successful experimental methodology for transformation and genome editing in winter wheat presented in this study will be of interest to the research community working with this crop. |
| first_indexed | 2024-03-10T09:27:37Z |
| format | Article |
| id | doaj.art-7c31388118cb409ea1a45a23f49eddcc |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T09:27:37Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-7c31388118cb409ea1a45a23f49eddcc2023-11-22T04:44:55ZengMDPI AGPlants2223-77472021-07-01107148110.3390/plants10071481Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat VarietiesFlorian Hahn0Laura Sanjurjo Loures1Caroline A. Sparks2Kostya Kanyuka3Vladimir Nekrasov4Plant Sciences Department, Rothamsted Research, Harpenden AL5 2JQ, UKPlant Sciences Department, Rothamsted Research, Harpenden AL5 2JQ, UKPlant Sciences Department, Rothamsted Research, Harpenden AL5 2JQ, UKDepartment of Biointeractions and Crop Protection, Rothamsted Research, Harpenden AL5 2JQ, UKPlant Sciences Department, Rothamsted Research, Harpenden AL5 2JQ, UKCRISPR/Cas technology has recently become the molecular tool of choice for gene function studies in plants as well as crop improvement. Wheat is a globally important staple crop with a well annotated genome and there is plenty of scope for improving its agriculturally important traits using genome editing technologies, such as CRISPR/Cas. As part of this study we targeted three different genes in hexaploid wheat <i>Triticum aestivum</i>: <i>TaBAK1-2</i> in the spring cultivar Cadenza as well as <i>Ta-eIF4E</i> and <i>Ta-eIF(iso)4E</i> in winter cultivars Cezanne, Goncourt and Prevert. Primary transgenic lines carrying CRISPR/Cas-induced indels were successfully generated for all targeted genes. While BAK1 is an important regulator of plant immunity and development, Ta-eIF4E and Ta-eIF(iso)4E act as susceptibility (S) factors required for plant viruses from the <i>Potyviridae</i> family to complete their life cycle. We anticipate the resultant homozygous <i>tabak1-2</i> mutant lines will facilitate studies on the involvement of BAK1 in immune responses in wheat, while <i>ta-eif4e</i> and <i>ta-eif(iso)4e</i> mutant lines have the potential to become a source of resistance to wheat spindle streak mosaic virus (WSSMV) and wheat yellow mosaic virus (WYMV), both of which are important pathogens of wheat. As winter wheat varieties are generally less amenable to genetic transformation, the successful experimental methodology for transformation and genome editing in winter wheat presented in this study will be of interest to the research community working with this crop.https://www.mdpi.com/2223-7747/10/7/1481CRISPRCas9plantgenome editingBAK1eIF4E |
| spellingShingle | Florian Hahn Laura Sanjurjo Loures Caroline A. Sparks Kostya Kanyuka Vladimir Nekrasov Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties Plants CRISPR Cas9 plant genome editing BAK1 eIF4E |
| title | Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties |
| title_full | Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties |
| title_fullStr | Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties |
| title_full_unstemmed | Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties |
| title_short | Efficient CRISPR/Cas-Mediated Targeted Mutagenesis in Spring and Winter Wheat Varieties |
| title_sort | efficient crispr cas mediated targeted mutagenesis in spring and winter wheat varieties |
| topic | CRISPR Cas9 plant genome editing BAK1 eIF4E |
| url | https://www.mdpi.com/2223-7747/10/7/1481 |
| work_keys_str_mv | AT florianhahn efficientcrisprcasmediatedtargetedmutagenesisinspringandwinterwheatvarieties AT laurasanjurjoloures efficientcrisprcasmediatedtargetedmutagenesisinspringandwinterwheatvarieties AT carolineasparks efficientcrisprcasmediatedtargetedmutagenesisinspringandwinterwheatvarieties AT kostyakanyuka efficientcrisprcasmediatedtargetedmutagenesisinspringandwinterwheatvarieties AT vladimirnekrasov efficientcrisprcasmediatedtargetedmutagenesisinspringandwinterwheatvarieties |