ABSTRACT
The Rickettsiales (Alphaproteobacteria) are obligate intracellular bacteria that colonize a wide range of eukaryotic hosts, including diverse metazoa and protists. Here, we characterize rickettsial endosymbionts discovered in the cytoplasm of the algivorous amoeboflagellates Viridiraptor invadens and Orciraptor agilis (Viridiraptoridae, Cercozoa, Rhizaria), supplying evidence of free-living, phagotrophic members of the Cercozoa serving as hosts for Rickettsiales. According to 16S rRNA gene phylogenies, the bacteria represent two closely related but distinct genotypes within a deep-branching rickettsial clade, which contains the genera "Candidatus Odyssella," "Candidatus Paracaedibacter," and "Candidatus Captivus." Using the full-cycle rRNA approach, we detected the novel bacteria in four of nine viridiraptorid strains tested. Furthermore, two specific oligonucleotide probes with a single-nucleotide-difference discriminated both bacterial genotypes by fluorescence in situ hybridization (FISH). We establish the candidate species "Candidatus Finniella inopinata" (found in Viridiraptor invadens) and "Candidatus Finniella lucida" (found in Orciraptor agilis) for the novel bacteria and propose a new, provisional family of Rickettsiales, "Candidatus Paracaedibacteraceae."
Subject(s)
Alphaproteobacteria/isolation & purification , Alphaproteobacteria/physiology , Cercozoa/microbiology , Symbiosis , Alphaproteobacteria/classification , Alphaproteobacteria/genetics , Cercozoa/physiology , Cytoplasm/microbiology , Molecular Sequence Data , PhylogenyABSTRACT
The genus Hyalodiscus Hertwig and Lesser, 1874 comprises naked freshwater amoebae with a unique set of characters, namely a vibrant orange-red colour, a discoid or fan-shaped morphology, and a characteristic rolling locomotion. Some species feed on the chloroplasts of green algae and were regarded as relatives of Vampyrella Cienkowski, 1865. However, because of striking morphological differences and the lack of molecular data, the exact relationship of Hyalodiscus to vampyrellids is still obscure. Here, I describe Hyalodiscus flabellus sp. nov., a bright orange, fan-shaped amoeba feeding on Oedogonium (Chlorophyceae), which likely is a close relative of the type species H. rubicundus Hertwig and Lesser, 1874. Sequence comparisons of the SSU rRNA gene revealed that H. flabellus belongs to a deep-branching, so far uncharacterised lineage of the order Vampyrellida (Rhizaria), here defined as family Hyalodiscidae POCHE, 1913. Based on these results, the systematic position of the genus Hyalodiscus could be finally clarified, accompanied by the revision of relevant diagnoses and a taxonomic summary. Furthermore, the work reports on endosymbiotic bacteria inhabiting the cytoplasm of H. flabellus, which were identified as "Candidatus Megaira polyxenophila" (Rickettsiales, Alphaproteobacteria) using the full cycle rRNA approach with newly designed FISH probes for this widespread endosymbiotic bacterium.