Molecular Identification of Rickettsial Endosymbionts in the Non-Phagotrophic Volvocalean Green Algae

Background: The order Rickettsiales comprises Gram-negative obligate intracellular bacteria (also called rickettsias) that are mainly associated with arthropod hosts. This group is medically important because it contains human-pathogenic species that cause dangerous diseases. Until now, there has been no report of non-phagotrophic photosynthetic eukaryotes, such as green plants, harboring rickettsias. Methodology/Principal Findings: We examined the bacterial endosymbionts of two freshwater volvocalean green algae: unicellular Carteria cerasiformis and colonial Pleodorina japonica. Epifluorescence microscopy using 49-6-deamidino-2phenylindole staining revealed the presence of endosymbionts in all C. cerasiformis NIES-425 cells, and demonstrated a positive correlation between host cell size and the number of endosymbionts. Strains both containing and lacking endosymbionts of C. cerasiformis (NIES-425 and NIES-424) showed a.10-fold increase in cell number and typical sigmoid growth curves over 192 h. A phylogenetic analysis of 16 S ribosomal (r)RNA gene sequences from the endosymbionts of C. cerasiformis and P. japonica demonstrated that they formed a robust clade (hydra group) with endosymbionts of various non-arthropod hosts within the family Rickettsiaceae. There were significantly fewer differences in the 16 S rRNA sequences of the rickettsiacean endosymbionts between C. cerasiformis and P. japonica than in the chloroplast 16 S rRNA or 18 S rRNA of the host volvocalean cells. Fluorescence in situ hybridization demonstrated the existence of the rickettsiacea

A gene transfer event suggests a long-term partnership between eustigmatophyte algae and a novel lineage of endosymbiotic bacteria

International audienceRickettsiales are obligate intracellular bacteria originally found in metazoans, but more recently recognized as widespread endosymbionts of various protists. One genus was detected also in several green algae, but reports on rickettsialean endosymbionts in other algal groups are lacking. Here we show that several distantly related eustigmatophytes (coccoid algae belonging to Ochrophyta, Stramenopiles) are infected by Candidatus Phycorickettsia gen. nov., a new member of the family Rickettsiaceae. The genome sequence of Ca. Phycorickettsia trachydisci sp. nov., an endosymbiont of Trachydiscus minutus CCALA 838, revealed genomic features (size, GC content, number of genes) typical for other Rickettsiales, but some unusual aspects of the gene content were noted. Specifically, Phycorickettsia lacks genes for several components of the respiration chain, haem biosynthesis pathway, or c-di-GMP-based signalling. On the other hand, it uniquely harbours a six-gene operon of enigmatic function that we recently reported from plastid genomes of two distantly related eustigmatophytes and from various non-rickettsialean bacteria. Strikingly, the eustigmatophyte operon is closely related to the one from Phycorickettsia, suggesting a gene transfer event between the endosymbiont and host lineages in early eustigmatophyte evolution. We hypothesize an important role of the operon in the physiology of Phycorickettsia infection and a long-term eustigmatophyte-Phycorickettsia coexistence

Biodiversity of “Non-model” Rickettsiales and Their Association with Aquatic Organisms

Representatives of the order Rickettsiales are obligate intracellular bacteria, traditionally including well-studied pathogens of humans and other vertebrates, such as Rickettsia, Orientia, Anaplasma, and Ehrlichia. In the last two decades, studies based on molecular characterization techniques have reshaped our view on the biodiversity of Rickettsiales, and the eukaryotic hosts they can exploit. Several new genera have been described in “traditional” Rickettsiales families Rickettsiaceae and Anaplasmataceae. Moreover, a new family, “Candidatus Midichloriaceae,” displaying diversity at least comparable with the “traditional” ones, has been described. Recent data show that the majority of extant genera of Rickettsiales (16 out of 24) are hosted exclusively (or at least partly) by aquatic organisms, such as protists (e.g., ciliates, amoebas, flagellates), and animals (e.g., cnidarians, mollusks, tunicates, leeches), while only ten genera include some terrestrial host, such as arthropods. Given the highly interwoven phylogenetic relationships among Rickettsiales hosted by aquatic and terrestrial hosts, it is likely that the ancestral host of Rickettsiales was an aquatic protist organism, and the adaptation to terrestrial environments occurred independently in several distinct sublineages at least six times. Newly discovered lineages of “non-model” Rickettsiales present unforeseen features for the order such as the presence of flagella. Future investigations on “non-model” Rickettsiales are crucial to gain insight on the evolutionary history of the whole group, including the origin of molecular mechanisms involved in pathogenesis for humans and vertebrates

The Hidden World of Rickettsiales Symbionts: “Candidatus Spectririckettsia obscura,” a Novel Bacterium Found in Brazilian and Indian Paramecium caudatum

Symbioses between bacteria and eukaryotes are widespread and may have significant impact on the evolutionary history of symbiotic partners. The order Rickettsiales is a lineage of intracellular Alphaproteobacteria characterized by an obligate association with a wide range of eukaryotic hosts, including several unicellular organisms, such as ciliates and amoebas. In this work, we characterized the Rickettsiales symbionts associated with two different genotypes of the freshwater ciliate Paramecium caudatum originated from freshwater environments in distant geographical areas. Phylogenetic analyses based on 16S rRNA gene showed that the two symbionts are closely related to each other (99.4% identity), belong to the family Rickettsiaceae, but are far-related with respect to previously characterized Rickettsiales. Consequently, they were assigned to a new species of a novel genus, namely “Candidatus Spectririckettsia obscura.” Screening on a database of short reads from 16S rRNA gene amplicon-based profiling studies confirmed that bacterial sequences related to the new symbiont are preferentially retrieved from freshwater environments, apparently with extremely scarce occurrence (< 0.1% positive samples). The present work provides new information on the still under-explored biodiversity of Rickettsiales, in particular those associated to ciliate host cells