PMC

Burkholderia hayleyella has a larger total population size with Dictyostelium discoideum, while B. agricolaris does not. We measured the total abundance of Burkholderia inoculated into soil microcosms as nonsymbiotic cells (Burkholderia treatment) or symbiotic within D. discoideum (Burkholderia + Dictyostelium treatment). We determined the abundance of each Burkholderia species in the entire soil microcosm, which included Burkholderia within D. discoideum and in the soil. (a) Abundance of Burkholderia in each treatment with isolates from Figure pooled by Burkholderia species. Filled circles are the mean Burkholderia abundance across all isolates, and open circles are the mean for each isolate. Timepoints are the same as in Figure . (b) Interaction plot for the two treatments of B. agricolaris and B. hayleyella. The endpoint of each line is the average Burkholderia abundance across all isolates and timepoints for the treatments in each Burkholderia species

Neighbor-joining tree of Burkholderia coxL (259 bp) clones from edge and canopy DNA extracts and coxL from reference cultures. Clone sequences are indicated with the site from which they were obtained. Sequences for Burkholderia isolates and clones (259 bp) were obtained using primers designed for this study. Sequences for the remaining isolates were obtained from genomic sequences or from amplifications described by King (). After alignment (ClustalX), only nongapped positions were considered for phylogenetic analysis. The tree was rooted with the form II coxL gene of Rhodobacter sphaeroides 2.4.1 (YP352939). Culture sequences were from the following organisms (accession numbers): Bradyrhizobium sp. ORS278 (YP_001207897), Burkholderia sp. I7 (GU128953), Burkholderia sp. DNBP6-1 (FJ466453), Burkholderia sp. DNBP16 (FJ466452), Burkholderia sp. DNBP18 (FJ466451), Burkholderia sp. DNBP20 (FJ466450), Burkholderia sp. PP51-2 (FJ713671), Burkholderia sp. Rim (GU128952), Carbophilus carboxidus (FJ466455), Mesorhizobium sp. KP12W (FJ152139), Shinella zoogloeoides strain FG1M5 (FJ152138), Stenotrophomonas strain LUP (AY307920), Oligotropha carboxidovorans (CAA57829), Silicibacter pomeroyi DSS-3 (YP_167616), Ruegeria sp. WHOI JT-08 (AAW88347), Burkholderia xenovorans LB400 (YP_558874), Burkholderia strain CP11 (FJ152141), Burkholderia strain PP52-1 (FJ152140), Burkholderia strain PO-04-17-38 (FJ713672), Burkholderia strain EB-2 (GU128954), Hydrogenophaga pseudoflava (HPU80806), Alkalilimnicola erlichei MLHE1 (YP_742401), and Rhodobacter sphaeroides form II coxL (YP_352939).

Homology analysis of Burkholderia sp. CQ001 and other Burkholderia strains (Burkholderia phenoliruptrix BR3459, Burkholderia xenovorans LB400, Burkholderia kuruiensis M130, and Burkholderia vietnamiensis G4).

In vitro PHA production by wild-type and PHA-related gene-depleted mutants of the Burkholderia symbiont. (A) Images of PHA-derived fluorescence under the fluorescence microscope. Phase-contrast images (left) and fluorescent images (right) of Burkholderia cells cultured in PB-G medium are stained with Nile blue A. (i) Wild type; (ii) ΔphaP1 mutant; (iii) ΔphaP2 mutant; (iv) ΔphaP3 mutant; (v) ΔphaP4 mutant; (vi) ΔphaR mutant; (vii) ΔphaC mutant; and (viii) ΔphaR/phaR Burkholderia mutant (scale bars, 0.5 μm). (B) Stained PHA granules of wild-type and mutant Burkholderia cells colonized in the midgut of fifth-instar nymph. (i) Wild type; (ii) ΔphaP1 mutant; (iii) ΔphaP2 mutant; (iv) ΔphaP3 mutant; (v) ΔphaP4 mutant; (vi) ΔphaR mutant; (vii) ΔphaC mutant; and (viii) ΔphaR/phaR Burkholderia mutant. (C) Flow cytometric histograms of PHA-derived fluorescence from Burkholderia cells cultured in YG medium (black), PB-G medium (red), and colonized symbiotic Burkholderia (blue). Burkholderia cells were stained with Nile blue A. (i) Wild type; (ii) ΔphaP1 mutant; (iii) ΔphaP2 mutant; (iv) ΔphaP3 mutant; (v) ΔphaP4 mutant; (vi) ΔphaR mutant; (vii) ΔphaC mutant; and (viii) ΔphaR/phaR Burkholderia strain.