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: | , , , , , |
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| Tipo de documento: | artigo |
| Estado: | Versão publicada |
| Data de publicação: | 2020 |
| País: | Colombia |
| Recursos: | Tecnológico de Antioquia |
| Repositório: | Repositorio Tdea |
| Idioma: | inglês |
| OAI Identifier: | oai:dspace.tdea.edu.co:tdea/2791 |
| Acesso em linha: | https://dspace.tdea.edu.co/handle/tdea/2791 |
| Access Level: | Acceso aberto |
| Palavra-chave: | Hermaphroditism Genome Editing Gene Editing Edición Génica Edição de Genes Actinidia spp Sex-determinant Plant transformation New breeding technologies |
| Resumo: | 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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