Symbiont location, host fitness, and possible coadaptation in a symbiosis between social amoebae and bacteria
- Sun Yat-sen University, China
- Washington University, United States
- Southern Illinois University, United States
Figures
Figure 1
Illustration of host-symbiont pairs used throughout the study.
D. discoideum clones were originally harvested from the wild in three different states: uninfected (indicated as naïve), or naturally infected with B. agricolaris or B. hayleyalla (indicated as …
Figure 2 with 1 supplement
Burkholderia Infections Differentially Alter Spore Viability According to Burkholderia Species and Host Background.
Total viable spores were determined for naïve and native hosts in their field harvested (a), cured (b), B. agricolaris lab-infected (c), and B. hayleyella lab-infected state (d). Four clones were …
Figure 2—figure supplement 1
Total Spore Number and Percent of Viable Spores for Burkholderia Infections in Diverse Host Backgrounds.
Total spores (top panel) and percent viable spores (bottom panel) were determined for naïve and native hosts in their field harvested (a), cured (b), B. agricolaris lab-infected (c), and B. …
Figure 3 with 1 supplement
Bacterial cells are found within Burkholderia exposed vegetative amoebae.
Transmission electron micrographs of vegetative amoebae show naïve and cured native amoebae with intracellular morphologies suggestive of active bacterial digestion with no evidence of intact …
Figure 3—figure supplement 1
Multi-lamellar bodies excreted by vegetative amoebae.
Transmission electron micrographs of vegetative amoebae identified multi-lamellar bodies inside uninfected amoebae, indicating successful digestion of bacterial food (a). Multi-lamellar bodies are …
Figure 4
Burkholderia is found abundantly in colonized vegetative amoebae.
Confocal imaging of fixed and stained vegetative amoebas show little to no intracellular bacteria in uninfected clones (a). However, abundant Burkholderia (Burkholderia-RFP shown in red) is found in …
Figure 5
Intracellular bacteria are retained in naïve migrating slugs exposed to Burkholderia and in native Burkholderia hosts.
Transmission electron micrographs of uninfected (a) show closely packed amoebae with internal structures reminiscent of previous bacterial digestion but without evidence of intact internal bacteria. …
Figure 6
Bacterial cells are retained in spore and stalk cells from Burkholderia-exposed hosts.
As visualized through transmission electron microscopy, (a) uninfected hosts form sturdy spores and stalk cells with no detectable bacteria. Spores and stalk cells retain intracellular bacteria in …
Figure 7
Burkholderia is retained in the sori of developed D. discoideum hosts and the percent of Burkholderia positive spores differs according to Burkholderia species.
Confocal images show no intra- or extracellular bacteria in uninfected spores (a) Abundant Burkholderia is seen in B. agricolaris (b) and B. hayleyella (c) hosts, with more infected spores seen for B…
Figure 8
Fruiting body morphology is differentially altered by Burkholderia colonization.
Macro photographs of fruiting bodies (a) show slightly different morphologies according to Burkholderia infection status. Sori measurements demonstrate that field collected native-hayleyella hosts …
Tables
Table 1
Dictyostelium discoideum clones used for this study.
Clones are divided into specific sets each with naive, native-ag, and native-ha field-collected counterparts. They were collected from Virginia, North Carolina, and Texas as indicated.
| Set | Clone | Status | Burkholderia | Location collected | Date collected | GPS coordinates |
|---|---|---|---|---|---|---|
| 1 | QS9 | Naïve | None | Virginia-Mt Lake Biological Station | Oct. 15 2000 | N 37° 21’, W 80° 31’ |
| QS70 | Native | B. agricolaris | Texas- Houston Arboretum | Jul. 15 2004 | N 29° 46’, W 95° 27’ | |
| QS11 | Native | B. hayleyella | Virginia-Mt Lake Biological Station | Oct. 15 2000 | N 37° 21’, W 80° 31’ | |
| 2 | QS18 | Naïve | None | Virginia-Mt Lake Biological Station | Oct. 15 2000 | N 37° 21’, W 80° 31’ |
| QS159 | Native | B. agricolaris | Virginia-Mt Lake Biological Station | May. 2008 | N 37° 21’, W 80° 31’ | |
| QS23 | Native | B. hayleyella | Virginia-Mt Lake Biological Station | Sep. 25 2000 | N 37° 21’, W 80° 31’ | |
| 3 | QS17 | Naïve | None | Virginia-Mt Lake Biological Station | Oct. 15 2000 | N 37° 21’, W 80° 31’ |
| QS161 | Native | B. agricolaris | Virginia-Mt Lake Biological Station | May. 2008 | N 37° 21’, W 80° 31’ | |
| QS22 | Native | B. hayleyella | Virginia-Mt Lake Biological Station | Sep. 25 2000 | N 37° 21’, W 80° 31’ | |
| 4 | QS6 | Naïve | None | Virginia-Mt Lake Biological Station | Sep. 25 2000 | N 37° 21’, W 80° 31’ |
| NC21 | Native | B. agricolaris | North Carolina-Little Butts Gap | Oct. 1988 | N 35° 46’, W 82° 20’ | |
| QS21 | Native | B. hayleyella | Virginia-Mt Lake Biological Station | Oct. 15 2000 | N 37° 21’, W 80° 31’ |
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strainbackground(Dictyostelium discoideum) | QS6 | Douglas et al., 2011, Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D.discoideum) | QS9 | Douglas et al., 2011, Brock et al., 2011 | Virginia-Mt Lake Biological Station | |
| Strain, strainbackground (D. discoideum) | QS17 | Douglas et al., 2011, Brock et al., 2011 | Virginia-MtLake Biological Station | |
| Strain, strainbackground (D. discoideum) | QS18 | Douglas et al., 2011, Brock et al., 2011 | Virginia-Mt Lake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | QS11 | Douglas et al., 2011, Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | QS21 | Douglas et al., 2011, Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | QS22 | Douglas et al., 2011, Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | QS23 | Douglas et al., 2011, Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | QS70 | Douglas et al., 2011 | Texas- HoustonArboretum | |
| Strain, strainbackground (D. discoideum) | QS159 | Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | QS161 | Brock et al., 2011 | Virginia-MtLake BiologicalStation | |
| Strain, strainbackground (D. discoideum) | NC21 | Francis and Eisenberg, 1993 | NC-Little ButtsGap | |
| Strain, strainbackground (Burkholderia hayleyella) | BhQS11 | Haselkorn et al., 2018 | isolated fromQS11 | |
| Strain, strainbackground (B. hayleyella) | BhQS21 | Haselkorn et al., 2018 | isolated fromQS21 | |
| Strain, strainbackground (B. hayleyella) | BhQS22 | Haselkorn et al., 2018 | isolated from QS22 | |
| Strain, strainbackground (B. hayleyella) | BhQS23 | Haselkorn et al., 2018 | isolated fromQS23 | |
| Strain, strainbackground (Burkholderia agricolaris) | BaQS70 | Haselkorn et al., 2018 | isolated fromQS70 | |
| Strain, strainbackground (B. agricolaris) | BaQS159 | Haselkorn et al., 2018 | isolated fromQS159 | |
| Strain, strainbackground (B. agricolaris) | BaQS161 | Haselkorn et al., 2018 | isolated fromQS161 | |
| Strain, strain background (B. agricolaris) | BaNC21 | Haselkorn et al., 2018 | isolated fromNC21 | |
| Strain, strain background (B. agricolaris) | BaQS70-RFP.1 | DiSalvo et al., 2015 | modified fromBaQS70 | |
| Strain, strain background (B. hayleyella) | BhQS11-RFP.2 | This paper | modified fromBaQS11 | |
| Strain, strainbackground (Klebsiella pneumoniae) | KpQS | Dictybase (http://dictybase.org/) | ||
| Strain, strainbackground (K. pneumoniae) | KpQS-GFP.1 | This paper | ||
| Recombinant DNA reagent | pmini-Tn7-KS-GFP | Teal et al., 2006 | ||
| Recombinant DNA reagent | pmini-Tn7-gat-P1-RFP | Su et al., 2014 |
Additional files
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Supplementary file 1
Statistical results of three fitness measures assayed for field-collected amoeba clones and after curing with antibiotics.
The three fitness measures were percent of spores that were viable, the total number of spores produced by a clone, and total viable spores. Total viable spores is the product of the other two measures. For each pairwise contrast, the essential difference in treatments is in boldface, and a treatment that is significantly higher than the other is marked with an asterisk and printed in red. Each of the fitness measures was analyzed with a set of Generalized Linear Mixed Models (GLMMs). This table gives the p-values for each question asked about main or interaction effects and the post hoc pairwise comparisons made, as relevant. Details about the statistical tests used can be found in the main text.
- https://doi.org/10.7554/eLife.42660.014
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Supplementary file 2
Statistical results of three fitness measures assayed for antibiotic-cured amoeba clones after experimental addition of Burkholderia.
he three fitness measures were again percent of spores that were viable, the total number of spores produced by a clone, and total viable spores. Total viable spores is the product of the other two measures. For each pairwise contrast, the essential difference in treatments is in boldface, and a treatment that is significantly higher than the other is marked with an asterisk and printed in red. Each of the fitness measures was analyzed with a set of Generalized Linear Mixed Models (GLMMs). This table gives the p-values for each question asked about main or interaction effects and the post hoc pairwise comparisons made, as relevant. Details about the statistical tests used can be found in the main text.
- https://doi.org/10.7554/eLife.42660.015
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Supplementary file 3
Statistical results for stalk morphology.
Each of the stalk measures was analyzed with a set of Generalized Linear Mixed Models (GLMMs). This table gives the p-values for each question asked about main or interaction effects and the post hoc pairwise comparisons made, as relevant. For each pairwise contrast, the essential difference in treatments is in boldface, and a treatment that is significantly higher than the other is marked with an asterisk and printed in red. Details about the statistical tests used can be found in the main text. One clone of each native type was tested: QS9 naïve; QS70 ag-infected; QS11 ha-infected.
- https://doi.org/10.7554/eLife.42660.016
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Supplementary file 4
Statistical results for sorus morphology.
Each of the spore measures was analyzed with a set of Generalized Linear Mixed Models (GLMMs). This table gives the p-values for each question asked about main or interaction effects and the post hoc pairwise comparisons made, as relevant. For each pairwise contrast, the essential difference in treatments is in boldface, and a treatment that is significantly higher than the other is marked with an asterisk and printed in red. Details about the statistical tests used can be found in the main text. One clone of each native type was tested: QS9 naïve; QS70 ag-infected; QS11 ha-infected.
- https://doi.org/10.7554/eLife.42660.017
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Transparent reporting form
- https://doi.org/10.7554/eLife.42660.018
