Rapid generation of a transgene-free powdery mildew resistant tomato by genome deletion

Genome editing has emerged as a technology with a potential to revolutionize plant breeding. In this study, we report on generating, in less than ten months, Tomelo, a non- transgenic tomato variety resistant to the powdery mildew fungal pathogen using the CRISPR/ Cas9 technology. We used wholegenome sequencing to show that Tomelo does not carry any foreign DNA sequences but only carries a deletion that is indistinguishable from naturally occurring mutations. We also present evidence for CRISPR/ Cas9 being a highly precise tool, as we did not detect off- target mutations in Tomelo. Using our pipeline, mutations can be readily introduced into elite or locally adapted tomato varieties in less than a year with relatively minimal effort and investment.