SlKIX8 and SlKIX9 are negative regulators of leaf and fruit growth in tomato

Abstract Plant organ size and shape are major agronomic traits that depend on cell division and expansion, which are both regulated by complex gene networks. In several eudicot species belonging to the rosid clade, organ growth is controlled by a repressor complex consisting of PEAPOD (PPD) and KINASE-INDUCIBLE DOMAIN INTERACTING (KIX) proteins. The role of these proteins in asterids, which together with the rosids constitute most of the core eudicot species, is still unknown. We used CRISPR-Cas9 genome editing to target SlKIX8 and SlKIX9 in the asterid model species tomato ( Solanum lycopersicum ) and analyzed loss-of-function phenotypes. We found that loss of function of SlKIX8 and SlKIX9 led to the production of enlarged, dome-shaped leaves and that these leaves exhibited increased expression of putative SlPPD target genes. Unexpectedly, kix8 kix9 mutants carried enlarged fruits with increased pericarp thickness due to cell expansion. At the molecular level, protein interaction assays indicated that SlKIX8 and SlKIX9 act as adaptors between the SlPPD and SlTOPLESS co-repressor proteins. Our results show that KIX8 and KIX9 are regulators of organ growth in asterids and can provide strategies to improve important traits in produce such as thickness of the fruit flesh. One sentence summary Two transcriptional repressors negatively regulate organ growth in tomato with loss-of-function lines producing enlarged fruits due to an appearance of more expanded cells in the fruit flesh.