Targeted mutagenesis using zinc-finger nucleases in perennial fruit trees

Reut Peer, Gil Rivlin, Sara Golobovitch, Moshe Lapidot, Amit Gal-On, Alexander Vainstein, Tzvi Tzfira, Moshe A. Flaishman

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

Genome editing is a powerful tool for precise gene manipulation in any organism; it has recently been shown to be of great value for annual plants. Classical breeding strategies using conventional cross-breeding and induced mutations have played an important role in the development of new cultivars in fruit trees. However, fruit-tree breeding is a lengthy process with many limitations. Efficient and widely applied methods for targeted modification of fruit-tree genomes are not yet available. In this study, transgenic apple and fig lines carrying a zinc-finger nuclease (ZFNs) under the control of a heat-shock promoter were developed. Editing of a mutated uidA gene, following expression of the ZFN genes by heat shock, was confirmed by GUS staining and PCR product sequencing. Finally, whole plants with a repaired uidA gene due to deletion of a stop codon were regenerated. The ZFN-mediated gene modifications were stable and passed onto regenerants from ZFN-treated tissue cultures. This is the first demonstration of efficient and precise genome editing, using ZFN at a specific genomic locus, in two different perennial fruit trees—apple and fig. We conclude that targeting a gene in apple or fig with a ZFN introduced by transient or stable transformation should allow knockout of a gene of interest. Using this technology for genome editing allows for marker gene-independent and antibiotic selection-free genome engineering with high precision in fruit trees to advance basic science as well as nontransgenic breeding programs.

Main conclusion: Targeting a gene in apple or fig with ZFN, introduced by transient or stable transformation, should allow genome editing with high precision to advance basic science and breeding programs.

Original languageEnglish
Pages (from-to)941-951
Number of pages11
JournalPlanta
Volume241
Issue number4
DOIs
StatePublished - 1 Apr 2015

Keywords

  • Apple (Malus domestica)
  • Fig (Ficus carica)
  • Perennial fruit trees
  • Targeted genome editing
  • Zinc-finger nuclease (ZFN)

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