Journal: Nature Communications

Article Title: Dual impact of elevated temperature on plant defence and bacterial virulence in Arabidopsis

doi: 10.1038/s41467-017-01674-2

Figure Lengend Snippet: Enhanced growth of Pst DC3000 in planta at 30 °C requires the type III secretion system and results in elevated levels of effector translocation. a Bacterial growth in plants ( n = 4) 3 days after syringe-infiltration with Pst DC3000. b Disease symptoms at three dpi for plants in ( a ). c Bacterial growth in plants ( n = 4) 3 days after syringe-infiltration with Pst DC3000, Pst DC3118 (coronatine-deficient mutant) or hrcC − (T3SS-deficient mutant) strains. d Disease symptoms three dpi for plants in ( c ). e Amount of cyclic AMP (cAMP) generated in temperature-acclimated plants syringe-infiltrated with Pst DC3000(P nptII ::avrPto-CyaA ) ( n = 4), Pst DC3000(P tac ::avrPtoB-CyaA ) ( n = 4), Pst DC3000(P tac ::hopU1-CyaA ) ( n = 6), Pst DC3000(P tac ::hopG1-CyaA ) ( n = 6) or hrcC − (P nptII ::avrPto-CyaA ) ( n = 4) strains. Tissue was collected at 4–6 hpi for quantification of cAMP, which was normalized by total protein. Higher levels of cAMP indicate more translocation of bacterial effectors. All data are representative of three independent experiments; graphical data are presented as the mean ± standard error of the mean (s.e.m.), with n = biological replicates. Letters indicate statistical significance based on a two-factor ANOVA with Tukey’s HSD post hoc analysis ( p < 0.05); samples sharing letters are not significantly different. Asterisks indicate statistical significance based on a Student’s t test (*** p < 0.001) of pairwise comparisons for each individual effector strain at 23 °C vs. 30 °C; ‘ns’ indicates no significance

Article Snippet: Statistical significance was determined using a Student’s t test (Excel) for pairwise comparisons, a one-way analysis of variance (ANOVA) followed by a Dunnett’s test (Prism 6, GraphPad Software, Inc.) for comparisons of multiple test samples to the same control, or by conducting a 2 × 2 factorial ANOVA followed by Tukey’s honest significant difference (HSD) test (RStudio ( https://www.rstudio.com/ ) for multi-variate analyses.

Techniques: Translocation Assay, Mutagenesis, Generated

Journal: Nature Communications

Article Title: Dual impact of elevated temperature on plant defence and bacterial virulence in Arabidopsis

doi: 10.1038/s41467-017-01674-2

Figure Lengend Snippet: Enhanced disease at elevated temperature corresponds to loss of SA biosynthesis. a SA marker gene expression ( n = 3) and ( b ) SA metabolite quantification ( n = 4) 24 h after vacuum-infiltration with mock or Pst DC3000. qPCR was used for gene expression analysis, with expression of ICS1 and PR1 normalized to the expression of PP2AA3 . SA and SAG metabolites were quantified using LCMS, converted to ng, normalized by sample fresh weight (FW) mass (g), and then combined and reported as total SA. c Bacterial growth in WT and ics1 mutant plants ( n = 4) at 1, 2 and 3 days after syringe-infiltration (dpi) with Pst DC3000. d Translocation of bacterial effectors in WT and ics1 mutant plants ( n = 4) syringe-infiltrated with Pst DC3000(P nptII ::avrPto-CyaA ). All data are representative of three independent experiments, and are presented as the mean ± s.e.m., with n = biological replicates. Letters indicate statistical significance based on a two-factor ANOVA with Tukey’s HSD post hoc analysis ( p < 0.05); samples sharing letters are not significantly different. Data for each time point in ( c ) were analysed separately

Article Snippet: Statistical significance was determined using a Student’s t test (Excel) for pairwise comparisons, a one-way analysis of variance (ANOVA) followed by a Dunnett’s test (Prism 6, GraphPad Software, Inc.) for comparisons of multiple test samples to the same control, or by conducting a 2 × 2 factorial ANOVA followed by Tukey’s honest significant difference (HSD) test (RStudio ( https://www.rstudio.com/ ) for multi-variate analyses.

Techniques: Marker, Gene Expression, Expressing, Mutagenesis, Translocation Assay

Journal: Nature Communications

Article Title: Dual impact of elevated temperature on plant defence and bacterial virulence in Arabidopsis

doi: 10.1038/s41467-017-01674-2

Figure Lengend Snippet: Induction of SA defence responses is compromised at 30 °C. a SA marker gene expression in plants ( n = 6) 24 h after spraying with mock or BTH. b SA metabolite quantification in plants ( n = 4) 48 h after spraying with mock or BTH. Gene expression and LCMS data were processed and analysed as described in Fig. . c Quantification and d representative images of callose accumulation from plants ( n = 6) 24 hpi with mock or BTH. Scale bar length represents 100 µm. All data are representative of three independent experiments. All graphical data are presented as the mean ± s.e.m., with n = biological replicates. Letters indicate statistical significance based on a two-factor ANOVA with Tukey’s HSD post hoc analysis ( p < 0.05); samples sharing letters are not significantly different

Article Snippet: Statistical significance was determined using a Student’s t test (Excel) for pairwise comparisons, a one-way analysis of variance (ANOVA) followed by a Dunnett’s test (Prism 6, GraphPad Software, Inc.) for comparisons of multiple test samples to the same control, or by conducting a 2 × 2 factorial ANOVA followed by Tukey’s honest significant difference (HSD) test (RStudio ( https://www.rstudio.com/ ) for multi-variate analyses.

Techniques: Marker, Gene Expression

Journal: Nature Communications

Article Title: Dual impact of elevated temperature on plant defence and bacterial virulence in Arabidopsis

doi: 10.1038/s41467-017-01674-2

Figure Lengend Snippet: Major SA antagonistic pathways are not responsible for enhanced susceptibility to Pst DC3000 at elevated temperature. a ET, c JA and e ABA marker gene expression in plants ( n = 3) 24 h after vacuum-infiltration with mock or Pst DC3000. Bacterial growth in WT and b ein3 eil1 , d myc2/3/4 , f aba2 and g camta2/3 mutant plants ( n = 4) 3 days after vacuum-infiltration with Pst DC3000. h SA metabolite quantification and i SA marker gene expression in WT and camta2/3 mutant plants ( n = 4) 3 days after vacuum-infiltration with mock or Pst DC3000. Gene expression and LCMS data were processed and analysed as described in Fig. . Data are presented as the mean ± s.e.m. with n = biological replicates and are representative of three independent experiments. Letters indicate statistical significance based on a two-factor ANOVA with Tukey’s HSD post hoc analysis ( p < 0.05); samples sharing letters are not significantly different. Data in h and i were analysed in two groups based on temperature

Article Snippet: Statistical significance was determined using a Student’s t test (Excel) for pairwise comparisons, a one-way analysis of variance (ANOVA) followed by a Dunnett’s test (Prism 6, GraphPad Software, Inc.) for comparisons of multiple test samples to the same control, or by conducting a 2 × 2 factorial ANOVA followed by Tukey’s honest significant difference (HSD) test (RStudio ( https://www.rstudio.com/ ) for multi-variate analyses.

Techniques: Marker, Gene Expression, Mutagenesis

Journal: Nature Communications

Article Title: Dual impact of elevated temperature on plant defence and bacterial virulence in Arabidopsis

doi: 10.1038/s41467-017-01674-2

Figure Lengend Snippet: BTH protection against Pst DC3000 at 30 °C requires NPR1 and TGAs but occurs without ICS1 and PR1 expression. a Bacterial growth in mock- or BTH-pre-treated plants ( n = 4) 3 days after vacuum-infiltration with Pst DC3000. b Disease symptoms three dpi in plants in a . c SA marker gene expression ( n = 3) and d SA metabolite quantification ( n = 4) 24 hpi of plants in a . Gene expression and LCMS data were processed and analysed as described in Fig. . Bacterial growth in mock- or BTH-pre-treated WT, e npr1 and f tga2/5/6 mutant plants ( n = 4) 3 days after vacuum-infiltration with Pst DC3000. g Translocation of bacterial effector proteins in plants ( n = 4) pre-treated with mock or BTH 24 h before syringe-infiltration with Pst DC3000(P nptII ::avrPto-CyaA ). Additional mock-treated plants ( n = 4 for each temperature) were infiltrated with hrcC − (P nptII ::avrPto-CyaA strains) as a negative control. Tissue was collected at 4 hpi for quantification of cAMP, which was normalized by total protein. All data are representative of three independent experiments; graphical data are presented as the mean ± s.e.m. with n = biological replicates. Letters indicate statistical significance based on a two-factor ANOVA with Tukey’s HSD post hoc analysis ( p < 0.05); samples sharing letters are not significantly different. Data in e and f were analysed in two groups based on temperature as indicated by the prime symbol (′). Symbols in g denote statistical significance based on a one-factor ANOVA with Dunnett’s post hoc analysis (***, ### p < 0.001) using the mock-treated sample at each temperature as the means for comparison

Article Snippet: Statistical significance was determined using a Student’s t test (Excel) for pairwise comparisons, a one-way analysis of variance (ANOVA) followed by a Dunnett’s test (Prism 6, GraphPad Software, Inc.) for comparisons of multiple test samples to the same control, or by conducting a 2 × 2 factorial ANOVA followed by Tukey’s honest significant difference (HSD) test (RStudio ( https://www.rstudio.com/ ) for multi-variate analyses.

Techniques: Expressing, Marker, Gene Expression, Mutagenesis, Translocation Assay, Negative Control, Comparison