title: Targeted gene mutation in tetraploid potato through transient TALEN expression in protoplasts.
abstract: 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.
authors.affiliation: Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden; Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp, Sweden.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Nicolia
authors.affiliation: Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp, Sweden.
authors.forename: Estelle
authors.initials: E
authors.lastname: Proux-Wéra
authors.affiliation: Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden.
authors.forename: Inger
authors.initials: I
authors.lastname: Åhman
authors.affiliation: Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden.
authors.forename: Nawaporn
authors.initials: N
authors.lastname: Onkokesung
authors.affiliation: Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden.
authors.forename: Mariette
authors.initials: M
authors.lastname: Andersson
authors.affiliation: Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp, Sweden. Electronic address: Erik.Andreasson@slu.se.
authors.forename: Erik
authors.initials: E
authors.lastname: Andreasson
authors.affiliation: Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden. Electronic address: Li-Hua.Zhu@slu.se.
authors.forename: Li-Hua
authors.initials: LH
authors.lastname: Zhu
mesh_terms: D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D020869:Gene Expression Profiling; D018745:Genome, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D000069600:Plant Breeding / Q000379:methods*; D011123:Polyploidy; D011523:Protoplasts / Q000378:metabolism*; D016415:Sequence Alignment; D011198:Solanum tuberosum / Q000235:genetics*; D014162:Transfection / Q000379:methods*
chemical_list: D004262:DNA Restriction Enzymes
keywords: Gene editing; Potato; Protoplast; Site-directed mutagenesis; TALEN; Transient gene expression