Nuclear localization of NPR1 is required for regulation of salicylate tolerance, isochorismate synthase 1 expression and salicylate accumulation in Arabidopsis

Abstract

Plant systemic acquired resistance (SAR) is a broad-spectrum immune response in which pathogen infection in local tissue induces resistance in systemic leaves. Activation of SAR requires the signal molecule salicylic acid (SA), which is primarily synthesized from chorismate via isochorismate through the action of isochorismate synthase 1 (ICS1) and a putative isochorismate pyruvate lyase. The Arabidopsis transcription coactivator NPR1 is a key regulator of SAR, which functions at multiple nodes in the SA signaling network. NPR1 not only acts downstream of SA to activate SAR, but also upstream of SA to suppress the expression of ICS1, thus inhibiting SA biosynthesis. NPR1 also positively regulates SA tolerance and plays a role in SA-mediated negative regulation of jasmonic acid (JA) signaling. The NPR1 protein contains a functional bipartite nuclear localization signal (NLS). It has been shown that the NLS and nuclear localization of NPR1 are required for activation of pathogenesis-related gene expression, whereas modulation of the crosstalk between SA- and JA-dependent defense pathways is mediated by cytosolic NPR1. In this study we used two transgenic lines, one expressing a mutated npr1 with a dysfunctional NLS and the other in which NPR1 nuclear localization can be induced by dexamethasone treatment, to test whether nuclear localization is required for other functions of NPR1. We found that prevention of NPR1 nuclear localization renders transgenic seedlings sensitive to the toxicity of high levels of SA and causes over-accumulation of ICS1 transcripts and SA in response to pathogen infection. Induction of NPR1 nuclear localization restores SA tolerance and normal accumulation of ICS1 transcripts and SA. These results indicate that the NLS and nuclear localization of NPR1 are required for regulation of SA tolerance, ICS1 expression and SA accumulation.