Virus induced transgene- and tissue-culture free heritable genome editing in tomato

Genome editing has emerged as a powerful tool for crop breeding. However, most commonly used approaches rely on tissue culture and the use of transgenic materials, which are time-consuming and labor-intensive, and often exhibit strong genotype dependency. Here we utilized tobacco rattle virus (TRV)-mediated ISYmu1 genome editing, coupled with in planta shoot regeneration, to achieve somatic and heritable genome editing in different tomato cultivars. By targeting the SlPDS gene, we successfully generated virus- and transgene-free homozygous mutant progeny in one generation. We further verified the method by generating somatic edits in SlDA1 , a gene with potential agronomic relevance. Given the wide host range of TRV and its compatibility with in planta shoot regeneration, our system could be broadly applicable for rapid, non-transgenic and heritable genome editing, thereby advancing functional genomics and crop improvement.