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. (C) 2015 Elsevier B.V. All rights reserved.

Keywords

Potato; Site directed mutagenesis; TALEN; Gene editing; Protoplast; Transient gene expression

Published in

Journal of Biotechnology
2015, volume: 204, pages: 17-24
Publisher: ELSEVIER SCIENCE BV

SLU Authors

• Nicolia, Alessandro

  • Department of Plant Protection Biology, Swedish University of Agricultural Sciences
    
  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Wera, Estelle

  • Department of Plant Protection Biology, Swedish University of Agricultural Sciences
    

• Åhman, Inger

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Onkokesung, Nawaporn

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Andersson, Mariette

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

• Andreasson, Erik

  • Department of Plant Protection Biology, Swedish University of Agricultural Sciences
    

• Zhu, Li-Hua

  • Department of Plant Breeding, Swedish University of Agricultural Sciences
    

Global goals (SDG)

SDG2 Zero hunger
SDG12 Responsible consumption and production

UKÄ Subject classification

Plant Biotechnology

Publication identifier

• DOI: https://doi.org/10.1016/j.jbiotec.2015.03.021

Permanent link to this page (URI)

https://res.slu.se/id/publ/68332