Transcriptomics and functional genomics of ROS-induced cell death regulation by RADICAL-INDUCED CELL DEATH1

Plant responses to changes in environmental conditions are mediated by a network of signaling events leading to downstream responses, including changes in gene expression and activation of cell death programs. Arabidopsis thaliana RADICAL-INDUCED CELL DEATH1 (RCD1) has been proposed to regulate plant stress responses by protein-protein interactions with transcription factors. Furthermore, the rcd1 mutant has defective control of cell death in response to apoplastic reactive oxygen species (ROS). Combining transcriptomic and functional genomics approaches we first used microarray analysis in a time series to study changes in gene expression after apoplastic ROS treatment in rcd1. To identify a core set of cell death regulated genes, RCD1-regulated genes were clustered together with other array experiments from plants undergoing cell death or treated with various pathogens, plant hormones or other chemicals. Subsequently, selected rcd1 double mutants were constructed to further define the genetic requirements for the execution of apoplastic ROS induced cell death. Through the genetic analysis we identified WRKY70 and SGT1b as cell death regulators functioning downstream of RCD1 and show that quantitative rather than qualitative differences in gene expression related to cell death appeared to better explain the outcome. Allocation of plant energy to defenses diverts resources from growth. Recently, a plant response termed stress-induced morphogenic response (SIMR) was proposed to regulate the balance between defense and growth. Using a rcd1 double mutant collection we show that SIMR is mostly independent of the classical plant defense signaling pathways and that the redox balance is involved in development of SIMR.
Author Summary Reactive oxygen species (ROS) are utilized in plants as signaling molecules to regulate development, stress responses and cell death. One extreme form of defense uses programmed cell death (PCD) in a scorched earth strategy to deliberately kill off cells invaded by a pathogen. Compared to animals, the regulation of plant PCD remains largely uncharacterized, particularly with regard to how ROS regulate changes in gene expression leading to PCD. Using comparative transcriptome analysis of mutants deficient in PCD regulation and publicly available cell death microarray data, we show that quantitative rather than qualitative differences in cell death gene expression appear to better explain the cell death response. In a genetic analysis with double mutants we also found the transcription factor WRKY70 and a component of ubiquitin mediated protein degradation, SGT1b, to be involved in regulation of ROS induced PCD.