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DOTAP, a lipidic transfection reagent, triggers Arabidopsis plant defense responses

  • Carolina Grandellis
  • Betiana S. Garavaglia
  • Natalia Gottig
  • Caroline Lonez
  • Jean-Marie Ruysschaert
  • Jorgelina Ottado
Original Article

Abstract

Main conclusion

DOTAP triggers Arabidopsis thaliana immunity and by priming the defense response is able to reduce bacterial pathogen attack.

DOTAP is a cationic lipid widely used as a liposomal transfection reagent and it has recently been identified as a strong activator of the innate immune system in animal cells. Plants are sessile organisms and unlike mammals, that have innate and acquired immunity, plants possess only innate immunity. A key feature of plant immunity is the ability to sense potentially dangerous signals, as it is the case for microbe-associated, pathogen-associated or damage-associated molecular patterns and by doing so, trigger an active defense response to cope with the perturbing stimulus. Here, we evaluated the effect of DOTAP in plant basal innate immunity. An initial plant defense response was induced by the cationic lipid DOTAP in the model plant Arabidopsis thaliana, assessed by callose deposition, reactive oxygen species production, and plant cell death. In addition, a proteomic analysis revealed that these responses are mirrored by changes in the plant proteome, such as up-regulation of proteins related to defense responses, including proteins involved in photorespiration, cysteine and oxylipin synthesis, and oxidative stress response; and down-regulation of enzymes related to photosynthesis. Furthermore, DOTAP was able to prime the defense response for later pathogenic challenges as in the case of the virulent bacterial pathogen Pseudomonas syringae pv. tomato. Disease outcome was diminished in DOTAP-pre-treated leaves and bacterial growth was reduced 100 times compared to mock leaves. Therefore, DOTAP may be considered a good candidate as an elicitor for the study of plant immunity.

Keywords

Cationic lipid DOTAP Arabidopsis thaliana Plant defense responses 

Abbreviations

BABA

β-Aminobutyric acid

BTH

Benzo(1,2,3)thiadiazole-7-carbothioic acid

DAB

3,3′-Diaminobenzidine

DMPC

1,2-Dimyristoyl-sn-glycero-3-phosphocholine

DOTAP

1,2-Dioleoyl-3-trimethylammonium-propane

Dpi

Days post-infiltration

ETI

Effector-triggered-immunity

Hpi

Hours post-infiltration

LPS

Lipopolysaccharide

INA

2,6-Dichloroisonicotinic acid

MAPK3

Mitogen-activated protein kinase 3

ΦNPQ

Non-photochemical quenching

ΦPSII

Operational efficiency of photosystem II

OPDA

Oxophytodienoic acid

PAL1

Phenylalanine ammonia-lyase 1

PAMP

Pathogen-associated molecular pattern

PR

Pathogenesis-related protein

PRR

Pattern recognition receptor

PTI

Pattern-triggered-immunity

RLK

Receptor-like kinase

ROS

Reactive oxygen species

Notes

Acknowledgements

We thank Rodrigo Vena for assistance with the microscopy confocal facility and Diego Aguirre for plant technical assistance. This work was supported by Grants from Argentine Federal Government and Fondo para la Investigación Científica y Tecnológica (PICT2013-0625 to JO) and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, Argentina) (PIP2014 to J. O). CG is a fellow and BSG, NG and JO are staff members of CONICET.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Carolina Grandellis
    • 1
  • Betiana S. Garavaglia
    • 1
  • Natalia Gottig
    • 1
  • Caroline Lonez
    • 2
  • Jean-Marie Ruysschaert
    • 2
  • Jorgelina Ottado
    • 1
  1. 1.Instituto de Biología Molecular Y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas (IBR-CONICET) and Facultad de Ciencias Bioquímicas y FarmacéuticasUniversidad Nacional de RosarioRosarioArgentina
  2. 2.Laboratory of Structure and Function of Biological MembranesUniversité Libre de BruxellesBrusselsBelgium

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