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番茄中同源染色体间的靶向重组用于精准育种。

Targeted recombination between homologous chromosomes for precise breeding in tomato.

机构信息

Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot 76100, Israel.

出版信息

Nat Commun. 2017 May 26;8:15605. doi: 10.1038/ncomms15605.

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.

摘要

同源重组(HR)在双亲染色体之间随机发生。在这里,我们报告了通过 CRISPR-Cas9 对体细胞诱导的 DNA 双链断裂(DSB)之间的同源染色体进行靶向重组。我们通过一个可视化和分子检测来证明这一点,其中 PSY1 基因的两个等位基因之间的 DSB 诱导导致黄色果实,野生型红色部分通过 HR 介导的 DSB 修复形成。我们还表明,在含有一个对 CRISPR 免疫和一个敏感的 psy1 等位基因的杂合子植物中,使用未断裂等位基因序列模板修复断裂等位基因是常见的结果。在另一个实验中,我们展示了 psy1 可食用番茄突变体和野生型 Solanum pimpinellifolium 之间的体细胞诱导 DSB 的证据,该 DSB 仅靶向 S. pimpinellifolium 等位基因。这使得可以对生殖传播的靶向体细胞 HR 事件进行特征描述,证明体细胞诱导的 DSB 可用于作物的精确育种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8d57/5458649/8020ab3a4819/ncomms15605-f1.jpg

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