PP2C phosphatase Pic14 negatively regulates tomato Pto/Prf‐triggered immunity by inhibiting MAPK activation

Abstract

SUMMARYType 2C protein phosphatases (PP2Cs) are emerging as important regulators of plant immune responses, although little is known about how they might impact nucleotide‐binding, leucine‐rich repeat (NLR)‐triggered immunity (NTI). We discovered that expression of the PP2C immunity‐associated candidate 14 gene (Pic14) is induced upon activation of the Pto/Prf‐mediated NTI response in tomato. Pto/Prf recognizes the effector AvrPto translocated into plant cells by the pathogen Pseudomonas syringae pv. tomato (Pst) and activate a MAPK cascade and other responses which together confer resistance to bacterial speck disease. Pic14 encodes a PP2C with an N‐terminal kinase‐interacting motif (KIM) and a C‐terminal phosphatase domain. Upon inoculation with Pst‐AvrPto, Pto/Prf‐expressing tomato plants with loss‐of‐function mutations in Pic14 developed less speck disease, specifically in older leaves, compared to wild‐type plants. Transient expression of Pic14 in leaves of Nicotiana benthamiana and tomato inhibited cell death typically induced by Pto/Prf and the MAPK cascade members M3Kα and Mkk2. The cell death‐suppressing activity of Pic14 was dependent on the KIM and the catalytic phosphatase domain. Pic14 inhibited M3Kα‐ and Mkk2‐mediated activation of immunity‐associated MAPKs and Pic14 was shown to be an active phosphatase that physically interacts with and dephosphorylates Mkk2 in a KIM‐dependent manner. Together, our results reveal Pic14 as an important negative regulator of Pto/Prf‐triggered immunity by interacting with and dephosphorylating Mkk2.