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Nitrate reductase is required for the transcriptional modulation and bactericidal activity of nitric oxide during the defense response of Arabidopsis thaliana against Pseudomonas syringae

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Abstract

Nitrate reductase (NR) has emerged as a potential NO source in plants. Indeed, the Arabidopsis thaliana NR double-deficient mutant (nia1 nia2) produces low NO and develops abnormal susceptibility to bacterial infection. We have employed quantitative real-time polymerase chain reactions to analyze the effects of NO gas on the expression of defense-related genes in wild-type and nia1 nia2 A. thaliana plants that were inoculated with an avirulent strain of Pseudomonas syringae pv. tomato. The pathogenesis-related gene 1 (PR1) was up-regulated by bacterial infection, and its expression was higher in the wild type than in nia1 nia2. Fumigation with NO attenuated the expression of PR1 and other salicylic acid-related genes in plants that had been inoculated with P. syringae. Nevertheless, NO inhibited the most intense bacterial growth and disease symptoms in nia1 nia2 leaves. The NO fumigation also directly modulated lignin biosynthesis-related gene expression (CAD1) and parts of the auxin (TIR1, ILL1, GH3) and ethylene (ACCS7) pathways, among other defense-related genes, and their modulation was more intense in the NR-deficient mutant. Pathogen inoculation induced delayed but intense H2O2 production in mutant leaves in comparison with the wild type. Hydrogen peroxide potentiated the microbicidal effects of NO against bacterial cultures. These results suggest that NO has a direct microbicidal effect in combination with H2O2 to allow for the attenuation of the SA-mediated defense response, thereby reducing the energy expenditure associated with defense-related gene transcription. Overall, these results highlight the importance of NR-dependent NO production in the establishment of disease resistance.

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Abbreviations

Arg:

l-Arginine

Cfu:

Colony forming units

HR:

Hypersensitive response

NO:

Nitric oxide

NOS:

Nitric oxide synthase

NR:

Nitrate reductase

PR:

Pathogenesis-related protein

Pst:

Pseudomonas syringae pv. tomato

ROS:

Reactive oxygen species

SA:

Salicylic acid

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Acknowledgments

This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, grant no. 473090/2011-2). S.C.V. and G.T.D. were supported by a student fellowship and H.C.O. by a research fellowship from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP). I.S. and M.G.A.V. are supported by a research fellowship from CNPq.

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  1. Departamento de Biologia Vegetal, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), CP 6109, Campinas, SP, 13083-970, Brazil

    Simone C. Vitor, Gustavo T. Duarte, Elzira E. Saviani, Michel G. A. Vincentz, Halley C. Oliveira & Ione Salgado

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  1. Simone C. Vitor
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  2. Gustavo T. Duarte
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  3. Elzira E. Saviani
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  4. Michel G. A. Vincentz
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  5. Halley C. Oliveira
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  6. Ione Salgado
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Correspondence to Ione Salgado.

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Vitor, S.C., Duarte, G.T., Saviani, E.E. et al. Nitrate reductase is required for the transcriptional modulation and bactericidal activity of nitric oxide during the defense response of Arabidopsis thaliana against Pseudomonas syringae . Planta 238, 475–486 (2013). https://doi.org/10.1007/s00425-013-1906-0

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