Fig 1.
Phylogenetic relationship of Rhabditophanes and selected Strongyloididae species based on(5, 25).
Only the tree topology is shown. The branch lengths are not informative.
Fig 2.
Life cycle and developmental stages of R. diutinus.
(A) previously described simple life cycle according to [25]. (B) Updated life cycle for R. diutinus. (C) J2A larva. (D) dauer larva. (E) J3 larva for comparison. (F) buccal plug (arrow) and (G) the uppermost of several intestinal plugs (star). (H) Front ends of J2A, lacking a buccal plug but featuring an ordinary narrow open mouth (arrows). All scale bars are 25μm.
Fig 3.
Recovery of dauers into adults in R. diutinus.
(A) Time course of dauer recovery. Images are taken from a time course experiment. Images are shown at 4 hours intervals from dauers (t = 0 hours) through to fully developed reproducing adults (t = 44 hours). (B-D) High magnification DIC images of the two molts, at 16 and 36 hours are shown. Scale bars indicate 100μm. The images shown here are a composite of the experiment and do not constitute a single worm as it develops.
Table 1.
Genes known to be involved in the dauer signaling pathway in C. elegans that are present within the published Rhabditophanes diutinus genome.
Table 2.
Genes present within 5 tested Strongyloididae genomes that are part of the C. elegans dauer signaling pathway.
Fig 4.
Heatmap of orthologs of known dauer genes in different life stages of R. diutinus.
Transcriptome profile of orthologs of known dauer genes in Adults, J2/J3 larvae, J2As and Dauers. Values shown are mean z score from all 3 biological replicates as replicates were determined to be similar (see S1 Fig). Z score indicates difference in expression between the different stages, with blue indicating a negative z score (-2 to 0) and decreased expression and red indicating a positive z score (0 to 2) and increased expression.
Table 3.
Significantly differentially expressed genes in R. diutinus between J2/J3s, J2As and dauers.
Fig 5.
Heatmap of known dauer genes present in all three species (R. diutinus, S. papillosus and C. elegans).
Z scores were determined and used to measure differences in expression for known dauer genes in all three species. Only genes that are present in all three species are shown. White indicates that the copy is not present within the genome, blue indicates a negative z score (-2 to 0) and decreased expression and red indicates a positive z score (0 to 2) and increased expression.
Table 4.
Dafachronic acid treatment prevents the formation of Dauer but not J2A larvae.
Fig 6.
Dauers survive longer than J2A or J2/J3 when on fresh foodless plates.
Survival curves of Dauers (dark blue line), J2As (green line) and J2/J3s (light blue line) when transferred onto fresh plates to induce starvation. Per treatment, 75 worms were picked in groups of 5 and checked daily for survival. The experiment was performed at three time points such that each time 25 worms per treatment were analyzed in parallel. The data points shown here are the daily mean of all 75 worms. Error bars are standard deviation.
Table 5.
No transgenerational effects of being a dauer or J2A larvae compared to a J2/J3 in terms of offspring produced and likelihood to become dauers in the next generation.
Fig 7.
R. diutinus alters its reproductive output in response to density.
R. diutinus J2s were picked either singularly (red), in groups of 5 (yellow) or 10 (green) onto plates and maintained for 120 hours or picked singularly onto plates and transferred daily to a new plate (blue). Offspring produced after 72, 96 and 120 hours was recorded. ANOVAs were performed between the different samples at each time point. For each density, at least 10 plates were picked, and the experiment was repeated three times. Data is shown as median (line), interquartile range (box) and range of values (whiskers). * indicates a statistically significant value of between 0.05 and 0.01, ** indicates a statistically significant value of between 0.01 and 0.001 and *** indicates a statistically significant value of lower than 0.001.