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

Plant J. 2024 Sep;119(6):2622-2637. doi: 10.1111/tpj.16937. Epub 2024 Jul 20.

Abstract

Type 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.

Keywords: Pseudomonas syringae; Solanum lycopersicum; plant immunity; protein phosphatase 2C; tomato.

MeSH terms

  • Gene Expression Regulation, Plant
  • MAP Kinase Signaling System
  • Mitogen-Activated Protein Kinases / genetics
  • Mitogen-Activated Protein Kinases / metabolism
  • Nicotiana / genetics
  • Nicotiana / immunology
  • Nicotiana / metabolism
  • Nicotiana / microbiology
  • Phosphoprotein Phosphatases / genetics
  • Phosphoprotein Phosphatases / metabolism
  • Plant Diseases* / genetics
  • Plant Diseases* / immunology
  • Plant Diseases* / microbiology
  • Plant Immunity* / genetics
  • Plant Proteins* / genetics
  • Plant Proteins* / metabolism
  • Plants, Genetically Modified
  • Protein Phosphatase 2C / genetics
  • Protein Phosphatase 2C / metabolism
  • Pseudomonas syringae* / physiology
  • Solanum lycopersicum* / genetics
  • Solanum lycopersicum* / immunology
  • Solanum lycopersicum* / metabolism
  • Solanum lycopersicum* / microbiology

Substances

  • Plant Proteins
  • Protein Phosphatase 2C
  • Phosphoprotein Phosphatases
  • Mitogen-Activated Protein Kinases