To diversify their genetic material, allowing adaptation to environmental disturbances and colonization of new ecological niches, bacteria use various evolutionary processes, including the acquisition of new genetic material by horizontal transfer mechanisms such as conjugation, transduction and transformation. Electrotransformation mediated by lightningrelated electrical phenomena may constitute an additional gene transfer mechanism occurring in nature. The presence in clouds of bacteria capable of forming ice nuclei that lead to precipitations and are involved in the triggering of lightning, such as the global phytopathogen Pseudomonas syringae, led us to postulate that natural electrotransformation in clouds may affect bacteria, by contributing to increase their adaptive potential. We first determined if the ice nucleator bacterium P. syringae could survive when in clouds and acquire exogenous genetic material through lightning shock-simulating in vitro electroporation. In comparison to two other bacteria, P. syringae appears to be best adapted for survival and for genetic electrotransformation under these conditions, which suggests that this bacterium would be able to survive and evolve whilst being transported in clouds. Secondly, we evaluated the impact of lightning shock-simulating in vitro electroporation on the survival, the electrotransformation potential and the diversity of bacteria collected from rain samples. These isolates better resisted lightning than the laboratory strains and some were able to electrotransform exogenous DNA. The rain bacteria we isolated were of different origins and were representative of life modes of the various sources of bacterial emissions on Earth. Our study suggests that bacteria aerosolized from diverse terrestrial ecosystems can spread to new habitats through clouds whilst also being able to acquire new genetic material via lightning-based electrotransformation, thereby potentially enhancing their genetic diversity. The final part of our work consisted of evaluating whether electrotransformation could be applied to the engineering of indigenous soil bacteria in order to develop a tool for the bioremediation of lindane, a once widely used pesticide. Optimized experiments revealed that both natural and electrotransformation contributed to the incorporation of a plasmid harboring a gene encoding the first lindane dechlorination steps by indigenous soil bacteria. In conclusion, we showed that natural electrotransformation mediated by electrical discharges such as those occurring in clouds or reaching soils can be involved in the horizontal gene transfer process among bacteria and, considering the importance of lightning worldwide, may play a role in the adaptation and evolution of these organisms.
| Identifer | oai:union.ndltd.org:CCSD/oai:tel.archives-ouvertes.fr:tel-01024190 |
| Date | 30 September 2013 |
| Creators | Blanchard, Laurine |
| Publisher | Ecole Centrale de Lyon |
| Source Sets | CCSD theses-EN-ligne, France |
| Language | English |
| Detected Language | English |
| Type | PhD thesis |
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