Targeted recombination between homologous chromosomes for precise breeding in tomato

Shdema Filler Hayut; Cathy Melamed Bessudo; Avraham Levy

doi:10.1038/ncomms15605

Targeted recombination between homologous chromosomes for precise breeding in tomato

Shdema Filler Hayut, Cathy Melamed Bessudo, Avraham Levy^*

^*Corresponding author for this work

Department of Plant and Environmental Sciences

Research output: Contribution to journal › Article › peer-review

160 Citations (Scopus)

Abstract

Homologous recombination (HR) between parental chromosomes occurs stochastically. Here, we report on targeted recombination between homologous chromosomes upon somatic induction of DNA double-strand breaks (DSBs) via CRISPR-Cas9. We demonstrate this via a visual and molecular assay whereby DSB induction between two alleles carrying different mutations in the PHYTOENE SYNTHASE (PSY1) gene results in yellow fruits with wild type red sectors forming via HR-mediated DSB repair. We also show that in heterozygote plants containing one psy1 allele immune and one sensitive to CRISPR, repair of the broken allele using the unbroken allele sequence template is a common outcome. In another assay, we show evidence of a somatically induced DSB in a cross between a psy1 edible tomato mutant and wild type Solanum pimpinellifolium, targeting only the S. pimpinellifolium allele. This enables characterization of germinally transmitted targeted somatic HR events, demonstrating that somatically induced DSBs can be exploited for precise breeding of crops.

Original language	English
Article number	15605
Number of pages	9
Journal	Nature Communications
Volume	8
DOIs	https://doi.org/10.1038/ncomms15605
Publication status	Published - 26 May 2017

Funding

We would like to thank Prof. Daniel Zamir, Prof. Yossi Hirschberg and Dr Itay Zemach (Hebrew University of Jerusalem) for providing the tomato mutant lines; Offir Lupo and Or Gross for the excellent technical help. This work was supported by the grants 1027/14 from the Israel Science Foundation (ISF); 15-01-0001 from the Israeli Ministry of Agriculture to A.A.L. S.F.H. performed most of the experimental work and took part in the planning of experiments and the writing of the manuscript; C.M.B. performed some of the experiments, provided experimental coaching and participated to the planning and the writing of the manuscript; A.A.L. took part in the planning of the experiments, in supervision of the work and in writing of the manuscript.

All Science Journal Classification (ASJC) codes

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General Physics and Astronomy

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Cite this

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abstract = "Homologous recombination (HR) between parental chromosomes occurs stochastically. Here, we report on targeted recombination between homologous chromosomes upon somatic induction of DNA double-strand breaks (DSBs) via CRISPR-Cas9. We demonstrate this via a visual and molecular assay whereby DSB induction between two alleles carrying different mutations in the PHYTOENE SYNTHASE (PSY1) gene results in yellow fruits with wild type red sectors forming via HR-mediated DSB repair. We also show that in heterozygote plants containing one psy1 allele immune and one sensitive to CRISPR, repair of the broken allele using the unbroken allele sequence template is a common outcome. In another assay, we show evidence of a somatically induced DSB in a cross between a psy1 edible tomato mutant and wild type Solanum pimpinellifolium, targeting only the S. pimpinellifolium allele. This enables characterization of germinally transmitted targeted somatic HR events, demonstrating that somatically induced DSBs can be exploited for precise breeding of crops.",

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Targeted recombination between homologous chromosomes for precise breeding in tomato

Abstract

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