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 language | American English |
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Pages (from-to) | 941-951 |
Number of pages | 11 |
Journal | Planta |
Volume | 241 |
Issue number | 4 |
DOIs | |
State | Published - Apr 2015 |
Bibliographical note
Funding Information:This study was supported by the Ministry of Agriculture, Bet Dagan, Israel.
Publisher Copyright:
© 2014, Springer-Verlag Berlin Heidelberg.
Keywords
- Apple (Malus domestica)
- Fig (Ficus carica)
- Perennial fruit trees
- Targeted genome editing
- Zinc-finger nuclease (ZFN)