Erwinia amylovora psychrotrophic adaptations: evidence of pathogenic potential and survival at temperate and low environmental temperatures

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Introduction

Material and Methods

Biosafety approval

Bacterial strains and culture conditions

Virulence

Growth curves

Siderophore detection

Motility assays in soft agar

EPS relative quantification

Biofilm quantification by a microtiter assay

Oxidative stress resistance

Microcosms preparation, monitoring of population dynamics and morphology analysis of starved cells

Statistical analysis

Results & Discussion

E. amylovora remains pathogenic even at 4 °C, with virulence, growth rates, siderophore secretion and motility decreasing in parallel to incubation temperature

EPS production, biofilm formation and oxidative stress resistance are enhanced at temperatures below the optimal for growth

Starvation at 14 °C favors the persistence in a culturable state, while the VBNC response is enhanced at 28 °C and, to a lesser extent, at 4 °C

Starvation induces morphological changes in a temperature-dependent manner

Supplemental Information

Virulence assays in immature pears (cv. Devoe) with the E. amylovora strain CFBP 1430 at 28 °C (A), 14 °C (B) and 4 °C (C)

Graphs show the percentage of symptomatic fruits over time. Representative pictures of fruits showing fire blight symptoms at the end of the experimental period (28 °C, 10 dpi; 14 °C, 15 dpi; 4 °C, 51 dpi) are represented besides the corresponding graph. Vertical lines indicate the SD.

DOI: 10.7717/peerj.3931/supp-1

Flow cytometry analysis of total and viable cell counts in E. amylovora starved cells at times 0 and 122 dpi

Strains Ea 1/79 (A), Ea 1189 (B), CFBP 1430 (C), NCPPB 2080 (D) and ATCC 49946. Vertical lanes indicate the SD. An ANOVA analysis of data revealed an effect of temperature on viability (p < 0.05) and total cell integrity (total cell counts) (p < 0.01).

DOI: 10.7717/peerj.3931/supp-2

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests.

Author Contributions

Ricardo D. Santander conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Elena G. Biosca conceived and designed the experiments, contributed reagents/materials/analysis tools, wrote the paper, reviewed drafts of the paper.

Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

The Universitat de València granted ethical and biosafety approval to carry out the study within its facilities.

Data Availability

The following information was supplied regarding data availability:

The raw data from representative experiments used to create the figures in the paper have been provided as Supplemental Files.

Funding

This work was supported by the project UV-INV-AE 112-66196 of the Universitat de València (UV), and also by the UV research support to the BACPLANT Laboratory. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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