Tesi di dottorato di ricerca
Endosymbiosis in Paramecium - Characterization of novel systems, transmission routes and the influence of abiotic stress factors
Settore scientifico disciplinare
Corso di studi
tutor Prof. Petroni, Giulio
relatore Prof. Berendonk, Thomas
relatore Prof. Berendonk, Thomas
Data inizio appello
Data di rilascio
Symbiosis between bacteria and single celled eukaryotes, such as ciliates and amoebae, is a widespread phenomenon. In the past years, the number of species characterizations of symbiotic bacteria, has constantly grown. Especially members of the order Rickettsiales (Alphaproteobacteria), which comprises obligate intracellular human and animal pathogens as well as other non-pathogenic intracellular species, got into the focus. However, biodiversity of both, protists and associated bacterial symbionts, is still largely underestimated. Thus, I hypothesised that different kind of bacteria occur naturally in the ciliate Paramecium, that paramecia can serve as potential reservoir for human and animal pathogens, and that the species distribution of Paramecium and its endosymbionts shows geographical patterns. In the present thesis, I provide data on the distribution and diversity of Paramecium species and their endosymbionts isolated from freshwater and brackish habitats from North and South America. The most frequent species were Paramecium biaurelia, Paramecium caudatum and Paramecium multimicronucleatum for the North American samples as well as P. multimicronucleatum and Paramecium calkinsi for the South American samples indicating geographical differences in the species distribution of Paramecium between the two continents. Monoclonal cultures were screened for the presence of endosymbionts by fluorescence in situ hybridization (FISH) using universal and specific probes. I detected bacterial endosymbionts belonging to different taxa of Rickettsiales sensu lato in Paramecium-cultures isolated from eight different habitats. Moreover, I characterized the four novel endosymbiont species “Candidatus Fokinia solitaria”, “Candidatus Fokinia cryptica”, “Candidatus Bealeia paramacronuclearis” and “Candidatus Jurandia parameciophila”, respectively, presenting detailed descriptions. Thus, I did not only detect various bacterial symbionts in different host species from two continents, but I also identified bacteria related to human and animal pathogens. Moreover, phylogenetic information provided by the four novel bacterial species helped me to revise the taxonomic organisation of the order Rickettsiales, which I split into the two orders Rickettsiales sensu stricto and Holosporales ord. nov.<br>The importance and severity of a symbiosis for its host can be estimated, among others, by studying the symbionts transmission modes. Most host-symbiont systems in nature show a mixed mode of transmission, even if one transmission type occurs only rarely. Moreover, evolution of exclusively vertically or horizontally transmitted symbionts is scarce. However, circumstances for horizontal transmission of many symbiont species so far described as vertically transmitted, are completely unknown and still need to be revealed. Therefore, I assumed that both, vector organisms and environmental stress, can trigger horizontal transmission. I performed different sets of infection experiments using vector organisms and environmental stress factors as driving forces to trigger horizontal transmission. In experiments focused on vector-induced transmission, I was not able to confirm vector properties of Holospora towards Caedibacter, as described towards food bacteria. However, I rarely observed vector-independent horizontal transmission of Caedibacter taeniospiralis towards symbiont-free Paramecium species. The here tested genotypes of C. taeniospiralis might still take advantage of additional horizontal transmission, even if it occurs rarely. In addition, I tested the effect of salinity and temperature stress on the transmission of the recently described bacterial symbionts “Candidatus Megaira polyxenophila” and “Ca. Bealeia paramacronuclearis” as potentially interesting representatives. The involved symbiont and host genotypes differed in their response to the induced environmental stress. Results of horizontal transmission turned out to be ambiguous. While temperature stress did not show any effect on transmission, salinity stress could possibly induce horizontal transmission of “Ca. Megaira polyxenophila” at slightly increased salinity concentrations in regard to normal conditions indicating the significance of environmental stress for the selection of the preferred transmission mode.
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