Isolation of Open Chromatin Identifies Regulators of Systemic Acquired Resistance

Abstract

Upon local infection, plants activate a systemic immune response called systemic acquired resistance (SAR). During SAR, systemic leaves become primed for the superinduction of defense genes upon reinfection. We used formaldehyde-assisted isolation of regulatory DNA elements coupled to next-generation sequencing to identify SAR regulators. Our bioinformatic analysis produced 10,129 priming-associated open chromatin sites in the 5' region of 3,025 genes in the systemic leaves of Arabidopsis (Arabidopsis thaliana) plants locally infected with Pseudomonas syringae pv. maculicola Whole transcriptome shotgun sequencing analysis of the systemic leaves after challenge enabled the identification of genes with priming-linked open chromatin before (contained in the formaldehyde-assisted isolation of regulatory DNA elements sequencing dataset) and enhanced expression after (included in the whole transcriptome shotgun sequencing dataset) the systemic challenge. Among them, Arabidopsis MILDEW RESISTANCE LOCUS O3 (MLO3) was identified as a previously unidentified positive regulator of SAR. Further in silico analysis disclosed two yet unknown cis-regulatory DNA elements in the 5' region of genes. The P-box was mainly associated with priming-responsive genes, whereas the C-box was mostly linked to challenge. We found that the P- or W-box, the latter recruiting WRKY transcription factors, or combinations of these boxes, characterize the 5' region of most primed genes. Therefore, this study provides a genome-wide record of genes with open and accessible chromatin during SAR and identifies MLO3 and two previously unidentified DNA boxes as likely regulators of this immune response.