Plant and soil microbe interactions in controlled conditions: rhizosphere protozoa and bacterial community structure
Plants influence the soil system by the large proportion of photosynthesized matters translocated to the roots and secreted into the soil. This root exudation provides an abundant energy source for rhizosphere living microorganisms. Plants are also strongly affected, positively and negatively, by the presence of soil microbiota, particularly bacteria, protozoa and fungi. Throughout the experiments conducted in this work, we aimed to better understand the influence of protozoa on plant growth. The first part of this work focused on the development of a microcosm method. Firstly, physical soil sterilization methods (autoclaving (A) gamma-ray irradiation (i) and both successively (AI)) were tested to eliminate the soil microbiota and their resistance form (spores and cysts). Although all sterilization methods tested were efficient to eliminate protozoa, AI was the only efficient method to eliminate aerobic heterotrophic cultivable bacteria without changing the soil pH. However the release of NH<sub>4</sub><sup>+</sup> in the soil after AI sterilization was higher than for other methods. Secondly, a procedure to re-inoculate the sterilized soil with a complex microbial community without protozoa was developed. The protozoa-free bacterial suspension was obtained from rhizosphere soil by subsequent filtering steps to exclude protozoa. The structure of bacterial communities characterised by 16SrDNA PCR-DGGE in the protozoa-free bacterial suspension was similar to that of the native soil. Diversity (Shannon) and evenness indexes increased with time in the sterile soil inoculated with the protozoa-free bacterial suspension. However the final bacterial community composition after 2 months of incubation in the re-inoculated soil presented a lower diversity as compared to the native soil. The second part of this work focused on the plant-microbiota interactions and on protozoa effects on plant growth. The microcosms developed in the first part of the work were re-inoculated with either sterile water or bacterial protozoa-free suspension or bacterial protozoa-free suspension and <i>Acanthamoeba castellanii</i> or with native soil suspension. The growth of <i>Arabidopsis thaliana</i> was clearly influenced by the inoculum and was particularly increased in presence of protozoa. Plants cultivated in presence of protozoa presented higher nitrogen content in leaves. The effect of leaf clipping (simulating herbivore damage) and nitrogen fertilization on soil microorganisms (bacteria, protozoa and nematodes) associated to the rhizosphere of barley was investigated in a pot experiment. The roots-shoots ratio decreased during the plant growth and was lower in the leaf clipping treatment. The abundance of bacteria was not significantly affected by leaf clipping and was higher in the high nitrogen-treatment. The abundance of bacterial-feeders (i.e. protozoa and nematodes) in the rhizosphere of 2, 4 and 6 weeks old plants was marginally affected by the nitrogen treatment as well as by leaf clipping. The role of protozoa in controlling the structure of bacterial community was investigated in the different experiment. The presence of protozoa did not change significantly the richness (numbers of bands) and the diversity (Shannon index) of the DNA-based DGGE fingerprints. The structure of the “total” bacterial communities was significantly changed in response to the functional group of protozoa (amoeba, ciliates and flagellates) inoculated as compared to the control (bacteria inoculum). The presence of protozoa did not change significantly the richness and the diversity of the RNA-based DGGE fingerprints. The structure of the active bacterial communities was significantly influenced by amoebas.
Thèse de doctorat : Université de Neuchâtel, 2008 ; Th. 2068
Type de publication
Resource Types::text::thesis::doctoral thesis
Dossier(s) à télécharger