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Bacteriophytochromes from Pseudomonas syringae pv. tomato DC3000 modulate the early stages of plant colonization during bacterial speck disease

  • Laura Moyano
  • Analía Carrau
  • Silvana Petrocelli
  • Ivana Kraiselburd
  • Wolfgang Gärtner
  • Elena G. OrellanoEmail author
Article
  • 21 Downloads

Abstra t

Living organisms have evolved the ability to perceive and respond to light of different wavelengths within the visible spectrum by the generation of photoreceptor proteins. Recent studies revealed the participation of these proteins in the virulence of plant pathogenic bacteria. Pseudomonas syringae pv. tomato DC3000 (Pto) is responsible for the bacterial speck, which affects tomato crops. Pto genome contains two genes encoding red/far-red light photoreceptors (BphP1: PSPTO_1902 and BphP2: PSPTO_2652). This work demonstrates the participation of Pto phytochromes and light in the bacterial physiology and during the interaction with tomato plants. We found that Pto phytochromes are implicated in the control of some features related with the bacteria capability to enter into the plant apoplast and cause bacterial speck disease, such as motility, biofilm formation, adhesion and emulsification capability. Red light and bacteriophytochromes are important during the early colonization stage of tomato phyllosphere, affecting Pto virulence. In addition, the development of disease symptoms in infiltrated leaflets is affected by light, which may be the consequence of type-two secretion system regulation.

Keywords

Bacteriophytochromes Light Virulence Pseudomonas syringae Plant colonization Bacterial speck disease 

Notes

Acknowledgments

We thank, Rodrigo Vena (IBR-CONICET) for microscopic analyses, the Statistics Department (Cátedra de Estadística Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario) for their assistance in the statistical analysis of the data and the staff from the English Department of the Facultad de Ciencias Bioquímicas y Farmacéuticas (UNR) for the language correction of the manuscript. The anti-flagellin antibodies were kindly provided by the laboratory of Dr. Eleonora García Véscovi.

Compliance with ethical standards

This research article is not submitted elsewhere for publication and this manuscript complies with the Ethical Rules applicable for this journal.

Competing interests

The authors declare that they have no competing interest.

Human and animal studies

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10658_2019_1918_Fig8_ESM.png (2 mb)
Fig. S1

Adhesion and biofilm formation by P. syringae pv. tomato DC3000 exposed to white light, red light and dark conditions. (A) Representative photograph of Pto WT, bphP1::Spr, cbphP1::Spr, bphP2::Gnr and cbphP2::Gnr adhered to the abaxial leaf surface and stained with Crystal violet dye. KB medium was taken as control. The order of inoculation is indicated at the left side of the panel. Dashed lines in the leaves indicate the inoculated area. (B) Representative photographs of biofilm formation assay. Pto strains were statically grown on glass flasks for 24 h at 28 °C. Biofilm formation levels on the air-liquid-glass interface were determined by Crystal violet staining. (D) Representative photographs of green fluorescent protein (GFP)-labeled bacteria were visualized under confocal laser scanning microscopy (CLSM) after three days of bacterial growth. Scale bars 50 μm. (PNG 2087 kb)

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High resolution image (TIF 7262 kb)
10658_2019_1918_Fig9_ESM.png (1.9 mb)
Fig S2

Biosurfactant production of P. syringae pv. tomato DC3000 exposed to white light, red light and dark conditions. Representative photograph of Pto WT, bphP1::Spr, cbphP1::Spr, bphP2::Gnr and cbphP2::Gnr biosurfactant production. (PNG 1948 kb)

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High resolution image (TIF 7504 kb)
10658_2019_1918_Fig10_ESM.png (1.1 mb)
Fig. S3

Effect of phytochrome genes disruption of P. syringae pv. tomato DC3000 and light on the epiphytic fitness. (A) Representative photograph of disease symptoms on tomato leaflets inoculated by dipping with Pto WT, bphP1::Spr, cbphP1::Spr, bphP2::Gnr and cbphP2::Gnr strains and exposed to white light, red light and darkness. (PNG 1130 kb)

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High resolution image (TIF 4616 kb)
10658_2019_1918_Fig11_ESM.png (407 kb)
Fig. S4

Bacterial growth curves in KB liquid medium. Growth curves of Pto WT, bphP1::Spr, cbphP1::Spr, bphP2::Gnr and cbphP2::Gnr strains in fresh KB medium considering the optical density at 600 nm (OD600) under (A) white light, (B) red light and (C) darkness, and the CFU/mL under (D) white light, (E) red light and (F) darkness, as a function of time. Data are represented as the mean ± standard error of two independent experiments. (PNG 407 kb)

10658_2019_1918_MOESM4_ESM.tif (1.1 mb)
High resolution image (TIF 1168 kb)
10658_2019_1918_Fig12_ESM.png (46 kb)
Fig. S5

Expression of bacteriophytochrome genes in WT and complemented strains. (A) Amplified products of the phytochrome genes by semiquantitative RT-PCR using RNA preparation from WT and complemented strains. As a control for constitutive bacterial expression a fragment of 16S rRNA was simultaneously amplified. (PNG 45 kb)

10658_2019_1918_MOESM5_ESM.tif (197 kb)
High resolution image (TIF 197 kb)
10658_2019_1918_MOESM6_ESM.docx (16 kb)
Appendix S1 . Statistical analysis for Figs. 1B, 5B and 6B. (DOCX 16 kb)

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Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2020

Authors and Affiliations

  • Laura Moyano
    • 1
    • 2
    • 3
  • Analía Carrau
    • 1
  • Silvana Petrocelli
    • 4
  • Ivana Kraiselburd
    • 1
  • Wolfgang Gärtner
    • 5
  • Elena G. Orellano
    • 1
    Email author
  1. 1.Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Ciencias Bioquímicas y FarmacéuticasUniversidad Nacional de RosarioRosarioArgentina
  2. 2.Facultad de Ciencias Exactas y Naturales, Departamento de Biodiversidad y Biología ExperimentalUniversidad de Buenos AiresBuenos AiresArgentina
  3. 3.Instituto de Biodiversidad y Biología Experimental (IBBEA)CONICET-Universidad de Buenos AiresBuenos AiresArgentina
  4. 4.Facultad de Ciencias Bioquímicas y FarmacéuticasUniversidad Nacional de RosarioRosarioArgentina
  5. 5.Max-Planck-Institute for Chemical Energy Conversion, D-45470 Mülheim, Germany; Institute for Analytical ChemistryLeipzigGermany

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