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

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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.

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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

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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.

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Correspondence to Jorgelina Ottado.

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The authors declare that they have no conflict of interest.

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Carolina Grandellis and Betiana S. Garavaglia contributed equally to this article.

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Grandellis, C., Garavaglia, B.S., Gottig, N. et al. DOTAP, a lipidic transfection reagent, triggers Arabidopsis plant defense responses. Planta 249, 469–480 (2019). https://doi.org/10.1007/s00425-018-3014-7

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  • DOI: https://doi.org/10.1007/s00425-018-3014-7

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