Fig. 1: C. elegans are attracted to PA14-produced ammonia. | Nature Communications

Fig. 1: C. elegans are attracted to PA14-produced ammonia.

From: Pseudomonas aeruginosa modulates both Caenorhabditis elegans attraction and pathogenesis by regulating nitrogen assimilation

Fig. 1

a Schematic of worm bacterial choice assay between PA14 and OP50. Worms were hatched and grown on OP50 for 2 days prior to bacterial choice assays. Worms were given 1 hour to chemotax to bacteria and paralyzed at first choice. ~30–100 worms were analyzed per plate, and precise numbers of worms used for each assay are noted in the source data file for each experiment. Created with BioRender.com. b Worm choice assay between PA14 WT/∆P11 and OP50. Choice index (CI) = (# on PA14–OP50)/(Total). p = 0.0117. c Choice assays between WT PA14 and ∆P11 with Wild type, che-2(e1033), che-1(p627), and odr-7(ky4) worm mutants (left) or odr-1(n1936), ceh-36(ky646), and ceh-37(ok642) (right). CI = (# on WT PA14–PA14 ∆P11)/(Total). d PA14 WT and ∆P11 ammonia production measured using an ammonia colorimetric assay. Colorimetric fluorescence intensity (ex = 360 nm/em = 450 nm) normalized to CFU and multiplied by 105. p = 0.0305. Chemical choice assays between water and ammonium acetate (e) and ammonium hydroxide (f) with Wild type, che-2(e1033), che-1(p627), and odr-7(ky4) worm mutants (left) or odr-1(n1936), ceh-36(ky646), and ceh-37(ok642) (right). CI = (# on chemical - # on water)/(# on chemical + # on water). g Schematic of bacterial choice assay with saturating concentrations of volatile ammonia. Bacteria spotted on NGM agar and chemical spotted on lid (5 spots of 10 µl chemical, 2.5 M each). Created with BioRender.com. h Addition of ammonia (ammonium acetate and ammonium hydroxide) to choice assay between PA14 WT and OP50. (CI) = (# on PA14–OP50)/(Total). i Addition of volatile ammonia (ammonium acetate and ammonium hydroxide) to choice assay between PA14 WT and ∆P11. CI = (# on PA14 WT– PA14 ∆P11)/(Total). Box Plots: center line = median, box range 25th–75th percentile, minimum/maximum denoted by whiskers, individual choice assay plates are represented by dots and are technical replicates (10 plates/technical replicates for (b, c (left), e (left), f (left), h, i) and 20 plates/technical replicates for c(right), e(right), f(right)). 3 Biological replicates were performed per choice assay and graphs represent one replicate. Bar graph error bars denote SD, center denotes mean, and each dot represents a biological replicate (3 total). One-Way ANOVA (c, e, f, h, i) Tukey’s multiple comparison test. Unpaired T-test, two-sided (b, d). *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, ns = not significant (Prism 9). For exact sample size (n), all biological replicates, and p values see source data file. Figure 1a, g were created with BioRender.com and released under a CC BY-NC-ND lisence.

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