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. 2014 Feb;80(3):878–885. doi: 10.1128/AEM.03310-13

FIG 3.

FIG 3

In situ identification of “Candidatus Hartigia pinicola” and “Candidatus Annandia pinicola” in different life stages (exulis and egg) of Pineus strobi. Bacterial symbionts were labeled by using symbiont-specific 16S and 23S rRNA-targeted oligonucleotide probes together with a general bacterial probe (see Table S2 in the supplemental material). Probes specific for phylotype 1 and phylotype 2 were labeled with Cy5 (blue) and Cy3 (red), respectively. The general bacterial probe (EUB338-I) was double labeled with Fluos (green). All probes were used simultaneously. The combined signals from the general and the symbiont-specific probes appear blue-green for “Candidatus Hartigia pinicola” (coccoid symbiont, phylotype 1) and yellow to orange for “Candidatus Annandia pinicola” (polymorphic symbiont, phylotype 2). (A) Bacteriocytes of P. strobi at the exulis life stage visualized by 16S rRNA-targeted symbiont-specific probes HarPi-265 and AnnPi-327. (B) Bacteriocytes in the exulis life stages visualized by 23S rRNA-targeted symbiont-specific probes HarPi-378 and AnnPi-1439. (C) Symbionts inside a P. strobi egg (probes HarPi-265 and AnnPi-1439). Bars, 10 μm.