Repressive C2H2 zinc finger ZAT proteins promote programmed cell death in the Arabidopsis columella root cap

Developmental programmed cell death (dPCD) controls a plethora of functions in plant growth and reproduction. In the root cap of Arabidopsis thaliana, dPCD functions to control organ size in balance with the continuous stem cell activity in the root meristem. Key regulators of root cap dPCD including SOMBRERO/ANAC033 (SMB) belong to the NAC family of transcription factors. Here we identify the C2H2 zinc finger protein ZAT14 as part of the gene regulatory network of root cap dPCD acting downstream of SMB. Similar to SMB, ZAT14 inducible misexpression leads to extensive ectopic cell death. Both thecanonical EARmotif and a conserved L-box motif of ZAT14 act as transcriptional repression motifs and are required to trigger cell death.While a single zat14 mutant does not show a cell death-related phenotype, a quintuple mutant knocking out five related ZAT paralogs shows a delayed onset of dPCD execution in the columella and the adjacent lateral root cap. While ZAT14 is co-expressed with established dPCD-associated genes, it does not activate their expression. Our results suggest that ZAT14 acts as a novel transcriptional repressor controlling a so far uncharacterized sub-section of the dPCD gene regulatory network active in specific root cap tissues.