RESUMEN
Resistance to Pseudomonas syringae pv. Maculicola 1 (RPM1) is a crucial immune receptor conferring plant enhanced resistance to pathogenic bacteria. RPM1-interacting protein 13 (RIN13) enhances RPM1-mediated disease resistance through interacting with the central domain of RPM1 in Arabidopsis, while the underlying mechanism remains elusive. Here, we report the subcellular localization and function of RIN13 using the Nicotiana benthamiana (N. benthamiana) transient expression system. Our results showed that RIN13 is exclusively localized in the nucleus, and RIN13 (231-300) fragment is responsible for its nuclear localization. Transient expression of RIN13 in N. benthamiana leaves can accelerate leaf senescence and cell death, and affect the activities of ROS-scavenging enzymes, and the C-terminus of RIN13 is crucial for its function. Furthermore, we identified a RIN13-interacting protein, Auxin Response Factor 1 (ARF1), and found that similar to RIN13, ARF1 can also promote leaf senescence and cell death. In addition, expression of RIN13 in N. benthamiana leaves can facilitate the translocation of ARF1 into the nucleus. Collectively, our study revealed a possible mechanism of RIN13 in accelerating leaf senescence and cell death by changing the subcellular localization of ARF1.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Núcleo Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción/metabolismo , Arabidopsis/citología , Arabidopsis/genética , Proteínas de Arabidopsis/análisis , Proteínas de Arabidopsis/genética , Muerte Celular , Núcleo Celular/genética , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/genética , Resistencia a la Enfermedad , Expresión Génica , Enfermedades de las Plantas/microbiología , Pseudomonas syringae/fisiología , Nicotiana/citología , Nicotiana/genética , Nicotiana/metabolismo , Factores de Transcripción/análisis , Factores de Transcripción/genéticaRESUMEN
Plants have evolved an innate immune system to protect themselves from pathogen invasion with the help of intracellular nucleotide-binding leucine-rich repeat (NLR) receptors, though the mechanisms remain largely undefined. RIN13 (RPM1-interacting protein 13) was previously reported to enhance disease resistance, and suppress RPM1 (a CNL-type NLR)-mediated hypersensitive response in Arabidopsis via an as yet unknown mechanism. Here, we show that RIN13 is a nuclear-localized protein, and functions therein. Overexpression of RIN13 leads to autoimmunity with high accumulation of salicylic acid (SA), constitutive expression of pathogenesis-related genes, enhanced resistance to a virulent pathogen, and dwarfism. In addition, genetic and transcriptome analyses show that SA-dependent and SA-independent pathways are both required for RIN13-mediated disease resistance, with the EDS1/PAD4 complex as an integration point. RIN13-induced dwarfism was rescued completely by either the pad4-1 or the eds1-2 mutant but partially by snc1-r1, a mutant of the TNL gene SNC1, suggesting the involvement of EDS1/PAD4 and SNC1 in RIN13 functioning. Furthermore, transient expression assays indicated that RIN13 promotes the nuclear accumulation of PAD4. Collectively, our study uncovered a signaling pathway whereby SNC1 and EDS1/PAD4 act together to modulate RIN13-triggered plant defense responses.