Targeted Mutagenesis of the Tomato Procera Gene Using TALENs

Transcription activator-like effector nucleases (TALENs) are efficient custom tools increasingly used for genome engineering. We successfully used TALENs under the control of an estrogen-inducible promoter for targeted mutagenesis in tomato (Solanum lycopersicum). Stable transgenic lines with TALENs targeting the negative regulator of gibberellin signaling PROCERA (PRO) were produced. Plants bearing pro mutations were regenerated from somatic cells of estrogen treated cotyledons. Six mono-allelic and one bi-allelic plants with deletions ranging in sizes from 1bp to 88bp at the TALEN target site were recovered. The mutant alleles were heritable and exhibited the expected Mendelian segregation pattern in the progeny of heterozygous plants. Homozygous mutant pro plants exhibited phenotypes consistent with increased gibberellin response. Our work demonstrates that TALENs can be used for targeted mutagenesis in tomato.