The YDA-MKK4/MKK5-MPK3/MPK6 Cascade Functions Downstream of the RGF1-RGI Ligand–Receptor Pair in Regulating Mitotic Activity in Root Apical Meristem

The mitotic activity of root apical meristem (RAM) is critical to primary root growth and development. Previous studies have identified the roles of ROOT GROWTH FACTOR 1 (RGF1), a peptide ligand, and its receptors, RGF1 INSENSITIVEs (RGIs), a clade of five leucine-rich-repeat receptor-like kinases, in promoting cell division in the RAM, which determines the primary root length. However, the downstream signaling components remain elusive. In this study, we identify a complete mitogen-activated protein kinase (MAPK or MPK) cascade, composed of YDA, MKK4/MKK5, and MPK3/MPK6, that functions downstream of the RGF1-RGI ligand-receptor pair. Similar to the rgi1/2/3/4/5 quintuple mutant, loss-of-function mutants of MPK3 and MPK6, MKK4 and MKK5, or YDA show a short-root phenotype, which is associated with reduced mitotic activity and lower expression of PLETHORA 1 (PLT1)/PLT2 in the RAM. Furthermore, MPK3/MPK6 activation in response to exogenous RGF1 treatment is impaired in the rgi1/2/3/4/5 quintuple, yda single, and mkk4 mkk5 double mutants. Epistatic analyses demonstrated that the expression of constitutively active MKK4, MKK5, or YDA driven by the RGI2 promoter can rescue the short-root phenotype of the rgi1/2/3/4/5 mutant. Taken together, these results suggest that the YDA-MKK4/MKK5-MPK3/MPK6 cascade functions downstream of the RGF1-RGI ligand-receptor pair and upstream of PLT1/PLT2 to modulate the stem cell population and primary root growth in Arabidopsis.