Targeted gene mutation in tetraploid potato through transient TALEN expression in protoplasts
| Autor: | Mariette Andersson, Alessandro Nicolia, Estelle Proux-Wéra, Erik Andreasson, Inger Åhman, Nawaporn Onkokesung, Li-Hua Zhu |
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| Rok vydání: | 2014 |
| Předmět: |
Molecular Sequence Data
Mutagenesis (molecular biology technique) Bioengineering Gene mutation Biology medicine.disease_cause Transfection Applied Microbiology and Biotechnology Loss of heterozygosity Polyploidy medicine Inbreeding depression Plant breeding Mutation frequency Gene Solanum tuberosum Genetics Mutation Base Sequence Gene Expression Profiling Protoplasts fungi food and beverages General Medicine DNA Restriction Enzymes Plant Breeding Mutagenesis Site-Directed Sequence Alignment Genome Plant Biotechnology |
| Zdroj: | Journal of biotechnology. 204 |
| ISSN: | 1873-4863 |
| Popis: | Potato is the third largest food crop in the world, however, the high degree of heterozygosity, the tetrasomic inheritance and severe inbreeding depression are major difficulties for conventional potato breeding. The rapid development of modern breeding methods offers new possibilities to enhance breeding efficiency and precise improvement of desirable traits. New site-directed mutagenesis techniques that can directly edit the target genes without any integration of recombinant DNA are especially favorable. Here we present a successful pipeline for site-directed mutagenesis in tetraploid potato through transient TALEN expression in protoplasts. The transfection efficiency of protoplasts was 38-39% and the site-directed mutation frequency was 7-8% with a few base deletions as the predominant type of mutation. Among the protoplast-derived calli, 11-13% showed mutations and a similar frequency (10%) was observed in the regenerated shoots. Our results indicate that the site-directed mutagenesis technology could be used as a new breeding method in potato as well as for functional analysis of important genes to promote sustainable potato production. |
| Databáze: | OpenAIRE |
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