Identification and characterization of SNO regulated genes (SRGs) in plant immunity
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A conspicuous feature of plants responding to pathogen invasion is the synthesis of nitric oxide (NO), a redox signal. NO regulates protein function by S-nitrosylation, the addition of an NO moiety to a cysteine thiol to form an S-nitrosothiol. A key theme of NO function is reprogramming plant immune-related gene expression. However, it is still not clear how the NO signal is translated into transcriptional changes. Here we explored the potential role of a sub-group of SNO Regulated Genes (SRGs) uncovered by global expression profiling. Firstly, transgenic plants containing the SRG1 or SRG3 promoter fused to glucuronidase gene GUS together with qRT-PCR assays confirmed that transcripts of SRGs could be induced by NO and pathogen challenge, suggesting that SRGs may be involved in NO signalling related to plant immunity. More importantly, transient and stable overexpression of SRG genes induced hypersensitive response (HR)-like cell death development, which is often associated with pathogen effector-triggered immunity. Furthermore, transgenic plants constitutively expressing SRG genes exhibited enhanced ROS accumulation, PR1 transcript accumulation, and increased resistance to Pseudomonas syringae (Pst) DC3000 compared with Col-0 wild type plants. In contrast, lines with T-DNA insertions into SRG genes exhibited susceptibility to Pst DC3000. These data suggested SRGs act as the positive regulators in plant immunity. In order to further explore how NO regulates these SRGs in plant immunity, we focused on SRG1 and found SRG1 could be S-nitrosylated in vitro and in vivo. Moreover, electrophoretic mobility shift assays showed SRG1 could bind to an AGT motif and the transcriptional activity was blunted in the presence of NO, suggesting that the DNA binding activity of SRG1 is redox-modulated. Further, a transient repression activity assay showed that SRG1 has repression activity and this activity was impaired in the gsnor1-3 mutant, which has a high S-nitrosothiols level. These data suggested NO could block SRG1 transcriptional activity in vitro and in vivo. Furthermore when the SRG1 overexpression line was crossed with gsnor1-3 the SRG1-mediated resistance related phenotypes were suppressed. These data demonstrated NO negatively regulates SRG1 transcriptional activity during plant immunity. SRG1 may therefore be an important regulator of NO signalling and subsequent regulate transcription during plant immunity. Additionally, NO may negatively feedback to inhibit transcriptional activity of SRG1 to control its repression activity, to enable the activation of plant immunity.