Fig. 5: Plasmid transfer under anaerobic conditions. | Nature Communications

Fig. 5: Plasmid transfer under anaerobic conditions.

From: Differential gut transmission of IncP plasmid clades involving hypervirulent Klebsiella pneumoniae reveals plasmid-specific ecological adaptation

Fig. 5: Plasmid transfer under anaerobic conditions.

a Log10-transformed conjugation efficiency of PTU-P1 (RP4-11, pMNCN064 and pYKBP039) and PTU-P2 (pKPC2, pVS0309 and pVS1035) plasmids from EcN to SGH10 measured under anaerobic conditions after 4 h of filter mating. The center line of the boxplot denotes the median, the box bounds indicate the 25th and 75th percentiles, and the whiskers extend to 1.5X the IQR. Each dot represents one biological replicate from a total of nine biological replicates for pKPC2 and RP4-11 or a total of twelve biological replicates for pMNCN064, pYKBP039, pVS0309 and pVS1035 from three independent experiments. Significant differences between PTU-P1 (n = 9 for RP4-11, n = 12 for pMNCN064 and pYKBP039) and PTU-P2 (n = 9 for pKPC2, n = 12 for pVS0309 and pVS1035) plasmids were determined using an unpaired t-test with Welch’s correction (two-sided) and are indicated by asterisks. The exact p value (PTU-P1 vs. PTU-P2): <0.0001. b Log10-transformed conjugation efficiency of pKPC2, RP4-11, pNDM1, and TP114 from EcN into SGH10 and SGH10ΔwcaJ measured under anaerobic conditions after 4 h of filter mating. The center line of the boxplot denotes the median, the box bounds indicate the 25th and 75th percentiles, and the whiskers extend to 1.5X the IQR. Each dot represents one biological replicate (n = 9) from three independent experiments. Significant differences between the anaerobic transfer efficiencies were determined using an unpaired t-test with Welch’s correction (two-sided) and are marked with asterisks. The exact p values (SGH10 vs. SGH10 ΔwcaJ): pKPC2 (0.9019); pNDM1 (0.9061); RP4-11 (0.0004); TP114 (0.0140). For comparison, aerobic data from Fig. 1a are shown in light overlay. Insets display the fold-change in conjugation efficiency into SGH10ΔwcaJ relative to SGH10 under respective aerobic and anaerobic conditions. c Mucoviscosity tests (supernatant-to-culture OD600 ratio) and d uronic acid quantification (µg/mL per 109 CFU) for SGH10 grown aerobically and anaerobically, with SGH10ΔwcaJ serving as the negative control. The center line of the boxplot in c denotes the median, the box bounds indicate the 25th and 75th percentiles, and the whiskers extend to 1.5X the IQR. Error bars in d represent mean ± SD. Each dot represents a biological replicate (n = 7 each for SGH10 aerobic and anaerobic; n = 5 SGH10ΔwcaJ) from three independent experiments. For both (c, d), one-way ANOVA with Tukey’s correction was used to assess statistical significance, with significant p values shown as asterisks. The exact p values (c, d): aerobic vs. anaerobic (<0.0001, 0.1408); aerobic vs. control (<0.0001, <0.0001); anaerobic vs. control (0.0673, <0.0001). e Log10-transformed conjugation efficiency of pKPC2, pNDM1, RP4-11 and TP114 from EcN into SGH10 (blue circles) and its mutants (SGH10ΔrmpA (orange triangles), SGH10ΔrmpC (pink squares), SGH10ΔrmpD (green plus signs), and SGH10ΔwcaJ (square with cross)) measured under aerobic conditions after 4 h of filter mating. The center line of the boxplot denotes the median, the box bounds indicate the 25th and 75th percentiles, and the whiskers extend to 1.5X the IQR. Each dot represents a biological replicate (n = 9) from three independent experiments. Statistical significance was assessed using one-way ANOVA (Dunnett’s test), with SGH10 as the reference group. Asterisks indicate statistical significance. The exact p values (SGH10 vs. SGH10ΔrmpA, SGH10 vs. SGH10ΔrmpC, SGH10 vs. SGH10ΔrmpD, SGH10 vs. SGH10ΔwcaJ): pKPC2 (<0.0001, 0.0282, 0.0044, <0.0001); pNDM1 (0.0107, 0.4998, 0.1765, <0.0001); RP4-11 (0.0007, 0.9986, 0.1992, <0.0001); TP114 (<0.0001, 0.0227, 0.6183, <0.0001). f Mucoviscosity tests (supernatant-to-culture OD600 ratio) and g uronic acid quantification (µg/mL per 109 CFU) for aerobically grown SGH10, SGH10ΔrmpA, SGH10ΔrmpC, SGH10ΔrmpD, and SGH10ΔwcaJ. The center line of the boxplot denotes the median, the box bounds indicate the 25th and 75th percentiles, and the whiskers extend to 1.5X the IQR. Each dot represents one biological replicate (n = 4) and the error bars represent mean ± SD. Statistical significance for (f, g) was assessed using one-way ANOVA (Dunnett’s test), with SGH10 as the reference group. Asterisks indicate statistical significance (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001), while “ns” denotes no significant difference. The exact p values for (f): SGH10 vs. SGH10ΔrmpA (<0.0001); SGH10 vs. SGH10ΔrmpC (0.9759); SGH10 vs. SGH10ΔrmpD (<0.0001); SGH10 vs. SGH10ΔwcaJ (<0.0001). The exact p values for (g): SGH10 vs. SGH10ΔrmpA (0.0165); SGH10 vs. SGH10ΔrmpC (0.0123); SGH10 vs. SGH10ΔrmpD (0.8007); SGH10 vs. SGH10ΔwcaJ (<0.0001).

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