Targeted Mutagenesis of the Female-Suppressor SyGI Gene in Tetraploid Kiwifruit by CRISPR/CAS9
Abstract: Kiwifruit belong to the genus Actinidia with 54 species apparently all functionally dioecious. The sex-determinants of the type XX/XY, with male heterogametic, operate independently of the ploidy level. Recently, the SyGI protein has been described as the suppressor of female development....
- Autores:
-
De Mori, Gloria
Zaina, Giusi
Franco Orozco, Bárbara
Testolin, Raffaele
De Paoli, Emanuele
Cipriani, Guido
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2020
- Institución:
- Tecnológico de Antioquia
- Repositorio:
- Repositorio Tdea
- Idioma:
- eng
- OAI Identifier:
- oai:dspace.tdea.edu.co:tdea/2791
- Acceso en línea:
- https://dspace.tdea.edu.co/handle/tdea/2791
- Palabra clave:
- Hermaphroditism
Genome Editing
Gene Editing
Edición Génica
Edição de Genes
Actinidia spp
Sex-determinant
Plant transformation
New breeding technologies
- Rights
- openAccess
- License
- https://creativecommons.org/licenses/by/4.0/
| Summary: | Abstract: Kiwifruit belong to the genus Actinidia with 54 species apparently all functionally dioecious. The sex-determinants of the type XX/XY, with male heterogametic, operate independently of the ploidy level. Recently, the SyGI protein has been described as the suppressor of female development. In the present study, we exploited the CRISPR/Cas9 technology by targeting two different sites in the SyGI gene in order to induce a stable gene knock-out in two tetraploid male accessions of Actinidia chinensis var. chinensis. The two genotypes showed a regenerative efficiency of 58% and 73%, respectively. Despite not yet being able to verify the phenotypic effects on the flower structure, due to the long time required by tissue-cultured kiwifruit plants to flower, we obtained two regenerated lines showing near fixation of a unique modification in their genome, resulting in both cases in the onset of a premature stop codon, which induces the putative gene knock-out. Evaluation of gRNA1 locus for both regenerated plantlets resulted in co-amplification of a minor variant differing from the target region for a single nucleotide. A genomic duplication of the region in proximity of the Y genomic region could be postulated. Keywords: Actinidia spp.; sex-determinant; hermaphroditism; plant transformation; genome editing; new breeding technologies (NBTs) |
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