Figure 3

In situ detection of Bacteroidales symbionts of gut protists of termites. (A-C) Ectosymbionts of Devescovina spp. in the gut of termite Neotermes koshunensis. (A) Ectosymbionts corresponding to the sequence NkFWB2-4 (labeled with 6FAM, stained in green) were associated with the host protist species represented by the sequence NkFWS (labeled with Texas Red, stained in red). (B) A mixture of Devescovina spp. cells was simultaneously hybridized with the sequence-specific probes for NkFWB2-4 (green) and NkD2-1 (red). (C) Ectosymbionts corresponding to the sequence NkD2-1 (red) were detected in the host protist species represented by the sequence Nk2 (green). Arrowheads in phase-contrast images of A-C indicate the stained Devescovina cells in colors corresponding to the probes, and the cells not detected by either probe are indicated by white arrowheads. (D-F) Detection of the ectosymbionts of protists in the genera Pyrsonympha and Dinenympha in the gut of R. speratus. (G) Detection of the ectosymbionts of Pyrsonympha sp.3 in the gut of H. sjoestedti. The upper panels in D-G show images obtained using sequence-specific probes labeled with 6FAM (green), and the middle panels show images obtained with the general bacterial probe (red). Insets in the upper panels of D-G are magnifications of the images indicated by squares. Arrows in panels D-G indicate typical ectosymbionts. Amorphous yellow signals in the upper panels and the corresponding signals in the middle panels were probably derived from autofluorescence of ingested wood particles. Scale bars: 50 μm (A-C, G) and 20 μm (D-F).