A Rapeseed WRKY Transcription Factor Phosphorylated by CPK Modulates Cell Death and Leaf Senescence by Regulating the Expression of ROS and SA-Synthesis-Related Genes

Abstract

Salicylic acid (SA) and reactive oxygen species (ROS) are two well-defined inducers of leaf senescence. Here, we identified a novel WRKY transcription factor gene WSR1 (WRKY regulating SA and ROS 1) in Brassica napus (rapeseed) in promoting SA and ROS production, which eventually led to leaf senescence thereafter. Its expression increased in senescing leaves. Ca2+-dependent protein kinase (CPK) 5 and -6 interacted with and phosphorylated BnaWSR1. Overexpression of phosphomimic BnaWSR1 (BnaWSR1ca) in rapeseed protoplasts elicited ROS production and cell death while its ectopic expression in Arabidopsis enhanced SA and ROS levels and, hence, accelerated leaf senescence. Furthermore, BnaWSR1ca activated the expression of Isochorismate Synthase 1 (ICS1), Respiratory Burst Oxidase Homologue (Rboh) D, and Senescence-Associated Gene 14 (SAG14). Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) assays demonstrated that BnaWSR1ca directly bound to promoter regions of ICS1, RbohD, and SAG14. These data have identified a CPK-WSR1 module that integrates SA and ROS to control cell death and leaf senescence.