Phylogenetic diversity of 'Endomicrobia' and their specific affiliation with termite gut flagellates

'Endomicrobia', a distinct and diverse group of uncultivated bacteria in the candidate phylum Termite Group I (TG-1), have been found exclusively in the gut of lower termites and wood-feeding cockroaches. In a previous study, we had demonstrated that the 'Endomicrobia' clones retrieved from Reticulitermes santonensis represent intracellular symbionts of the two major gut flagellates of this termite. Here, we document that 'Endomicrobia' are present also in many other gut flagellates of lower termites. Phylogeny and host specificity of 'Endomicrobia' were investigated by cloning and sequencing of the small subunit rRNA genes of the flagellate and the symbionts, which originated from suspensions of individual flagellates isolated by micropipette. Each flagellate harboured a distinct phylogenetic lineage of 'Endomicrobia'. The results of fluorescent in situ hybridization with 'Endomicrobia'-specific oligonucleotide probes corroborated that 'Endomicrobia' are intracellular symbionts specifically affiliated with their flagellate hosts. Interestingly, the 'Endomicrobia' sequences obtained from flagellates belonging to the genus Trichonympha formed a monophyletic group, suggesting co-speciation between symbiont and host.