Efficient tobacco rattle virus-induced gene editing in tomato mediated by the CRISPR/Cas9 system

Plant virus-based sgRNA delivery strategy has been widely applied for efficient genome editing across various plant species, leveraging its significant advantages in the rapid expression and expansion of sgRNA through virus replication and movement. However, the efficacy of the virus-induced gene editing (VIGE) tool in tomato has yet to be explored. In this paper, we established a TRV-mediated CRISPR/Cas9 genome editing system in the somatic cells of tomato, reporting the validation of VIGE and evaluating the mutagenesis efficiency in both tomato leaves and fruits using high-throughput sequencing. The results demonstrated an approximate 65% efficiency of VIGE in tomato leaves for the selected target genes, with VIGE efficiency reaching up to 50% in tomato fruits. This research not only introduces an efficient tool for reverse genetics but also reveals substantial potential of VIGE in surpassing traditional tissue culture techniques for creating heritable mutations in tomato. Establishment of the TRV-mediated CRISPR/Cas9 genome editing system in tomato somatic cells. Efficiency of TRV-induced gene editing in tomato leaves was approximately 65%. Efficiency of TRV-induced gene editing in tomato fruits was up to 50%. VIGE is useful for rapid gene function studies in tomato without conventional tissue culture. image