Suppression of class I compensated cell enlargement by xs2 mutation is mediated by salicylic acid signaling.

The regulation of leaf size has been studied for decades. Enhancement of post-mitotic cell expansion triggered by impaired cell proliferation in Arabidopsis is an important process for leaf size regulation, and is known as compensation. This suggests a key interaction between cell proliferation and cell expansion during leaf development. Several studies have highlighted the impact of this integration mechanism on leaf size determination; however, the molecular basis of compensation remains largely unknown. Previously, we identified extra-small sisters (xs) mutants which can suppress compensated cell enlargement (CCE) via a specific defect in cell expansion within the compensation-exhibiting mutant, angustifolia3 (an3). Here we revealed that one of the xs mutants, namely xs2, can suppress CCE not only in an3 but also in other compensation-exhibiting mutants erecta (er) and fugu2. Molecular cloning of XS2 identified a deleterious mutation in CATION CALCIUM EXCHANGER 4 (CCX4). Phytohormone measurement and expression analysis revealed that xs2 shows hyper activation of the salicylic acid (SA) response pathway, where activation of SA response can suppress CCE in compensation mutants. All together, these results highlight the regulatory connection which coordinates compensation and SA response. Author summary Leaves are determinate organ and size of leaves are determined by intrinsic and extrinsic cues. Cell proliferation and post-mitotic cell expansion should be coordinated during leaf morphogenesis to develop appropriate size depending on its developmental programs. Recent studies highlighted the existence of integrated mechanism which coordinates cell proliferation and cell expansion during leaf development. Compensation, which is enhanced post-mitotic cell expansion accompanied by a significant decrease in cell number during leaf organogenesis, is one of the clues for such coordination. However, the molecular mechanisms linking cell proliferation and cell expansion are still poorly understood. Previously, we reported extra-small sisters 2 (xs2) mutation caused specific defect in cell expansion and it suppressed increased post-mitotic cell enlargement in an3 mutant, which exhibits typical compensation. Here we identify the affected gene of xs2 mutant encodes a member of cation calcium exchanger which is believed to be involved in cation homeostasis within cells. Loss of function of this protein causes hyper accumulation of salicylic acid (SA) and increased expression of pathogen related genes. Physiological and genetic studies revealed activated SA signal transduction reduced cell size. It suppressed post-mitotic cell expansion in several compensation mutants not only an3 but partially suppressed in another type of compensation mutant which increases size of mitotic cells. This finding suggests post-mitotic cell expansion pathway is regulated in common by SA-dependent signaling and by compensation signaling during leaf development.