Le Miroir des sports : publication hebdomadaire illustrée

17 août 19371937/08/17 (N964)-1937/08/17

Additional file 1: Figure S1. of Comparative genome and phenotypic analysis of three Clostridioides difficile strains isolated from a single patient provide insight into multiple infection of C. difficile

Growth curves in BHIS of DSM 27638, DSM 27639 and DSM 27640. The isolates were grown in brain heart infusion medium containing 0.5% (w/v) yeast extract und 0.03% (w/v) L-cysteine. All isolates have the same growth rate under laboratory conditions as shown as the means of three replicates with standard deviation. (DOCX 55 kb

The transcriptional regulator-encoding gene <i>MTF1</i> of <i>Verticillium dahliae</i> is required for formation of microsclerotia at levels comparable to the wild-type.

(a, b) The ex-planta phenotype of the V. dahliae MTF1 deletion strain (ΔMTF1) was compared with the JR2 wild-type (WT) and the MTF1 complementation strain (MTF1-C). 50,000 spores of respective strains were point-inoculated onto plates containing simulated xylem medium (SXM), potato dextrose medium (PDM) or Czapek-Dox medium (CDM) and incubated at 25°C for 10 days. (a) Bottom view scans depict fewer microsclerotia formed by ΔMTF1 compared with the wild-type and MTF1-C on all tested media. For CDM + cellulose plates, close-up views, cross-sections of colony centers and microscopic images of fungal material from the colony centers are shown (black scale in upper two rows = 500 μm, orange scale in lower row = 50 μm). (b) Melanization of fungal colonies grown on CDM + cellulose was quantified as a measure of the number of microsclerotia formed. Pictures were taken from top view scans of the colonies after removal of aerial mycelium. The brightness factor of three colonies per transformant (n = 1) was calculated using the ROI Manager of ImageJ software. Three independent experiments were performed. Means ± SD are shown. Significant differences from wild-type were determined using t-tests. Melanization of ΔMTF1 was reduced to approximately 60% of wild-type levels (****, P MTF1-C (ns, not significant). (c) Expression levels of VTA1, which encodes the microsclerotia melanization-controlling transcription factor Vta1, were compared between wild-type, MTF1 deletion and complementation strains. RNA was extracted from mycelia grown in liquid SXM for five days and subsequently in xylem sap for 8 h. Means of three biological replicates normalized to transcript levels of wild-type and references H2A and EIF2B with error bars representing the SE of the means are shown. VTA1 expression was reduced to 60% in ΔMTF1 (*, P MTF1-C (calculated using t-tests).</p

<i>Verticillium dahliae</i> Vta3 does not affect the expression of effector genes <i>AVE1</i> and <i>SCP7</i>.

(DOCX)</p

Tomato pathogenesis-related proteins (PR) gene expression is activated after treatment with spores of <i>Verticillium dahliae</i> wild-type (WT) but not <i>VTA3</i> deletion strain (Δ<i>VTA3</i>).

(DOCX)</p

Proteins interacting with Vta3-GFP during vegetative growth.

(DOCX)</p

Expression of fungal effector genes and pathogenesis-related protein (PR) genes in tomato plants is increased in the presence of <i>Verticillium dahliae MTF1</i>.

Transcript levels were analyzed by reverse transcription-quantitative PCR and gene expression levels in wild-type were set to one. Significant differences from wild-type (WT) were determined using t-tests. Means of three independent experiments are shown with error bars representing the SE of the means. (a,c) Strains were cultured in extracted tomato xylem sap for 8 h following preculture in simulated xylem medium. Means of three biological replicates normalized to the transcript levels of reference genes H2A and EIF2B are shown. (a) In the absence of MTF1 (ΔMTF1), expression of the virulence factor Ave1-encoding gene was reduced by almost 80% compared with wild-type and the MTF1 complementation strain (MTF1-C) (*, P V. dahliae JR2 wild-type, ΔMTF1 or MTF1-C spores. Water-treated plants (mock) served as controls. Hypocotyls from 13 to 15 plants per treatment were pooled (n = 1). Means of six biological replicates normalized to the transcript levels of reference genes EF1α and αTUB are shown. The expression levels of PR1, PR1a, PR3 and PR5 did not differ significantly between wild-type and MTF1-C-infected plants. In plants treated with water or ΔMTF1, the expression levels of PR genes were significantly lower (**, P P P MTF1 to 40% of the wild-type level.</p

<i>Verticillium dahliae</i> Mtf1 does not affect <i>CMR1</i> expression.

(DOCX)</p

Plasmids used in this study.

(DOCX)</p

Verification of <i>Verticillium dahliae ELV1</i> deletion and complementation strains.

(DOCX)</p