Lignin is the second most abundant biopolymer on earth and is found in the wood (xylem) of vascular land plants. To transport the hydro-mineral sap, xylem forms specialized conduit cells, called tracheary elements (TEs), which are hollow dead cylinders reinforced with lateral secondary cell walls (SCW). These SCWs incorporate lignin to gain mechanical strength, water impermeability and resistance against pathogens. The aim of this thesis is to understand the spatio-temporal deposition of lignin during TE differentiation and the relationship with its neighbouring cells. In vitro TE differentiating cell cultures of Zinnia elegans and Arabidopsis thaliana are ideal tools to study this process: cells differentiate simultaneously into 30-50% TEs while the rest remain parenchymatic (non-TEs). Live-cell imaging of such TEs indicated that lignification occurs after programmed cell death (PCD), in a non-cell autonomous manner, in which the non-TEs provide the lignin monomers. This thesis confirms that lignification occurs and continues long after TE PCD in both in vitro TE cultures and whole plants using biochemical, pharmacological and cytological methods. The cooperative supply of lignin monomers by the non-TEs was demonstrated by using Zinnia and Arabidopsis in vitro TE cultures. Inhibitor experiments revealed further that the non-TEs supply reactive oxygen species (ROS) to TEs and that ROS are required for TE post-mortem lignification. Characterization of the non-TEs showed an enlarged nucleus with increased DNA content, thus indicating that non-TEs are in fact endoreplicated xylem parenchyma cells (XP). The cooperative lignification was confirmed in whole plants by using knock-out mutants in a lignin monomer synthesis gene, which exhibit reduced TE lignification. The XP specific complementation of these mutants led to nearly completely rescuing the TE lignin reduction. Using microscopic techniques, the spatial distribution of lignin was analyzed in TEs from in vitro cultures and whole plants, revealing that lignification is restricted to TE SCWs in both protoxylem and metaxylem. These specific deposition domains were established by phenoloxidases, i.e. laccases localized to SCWs and peroxidases, present in SCWs and the apoplastic space. Laccases were cell-autonomously produced by developing TEs, indicating that the deposition domains are defined before PCD. Altogether, these results highlight that the hydro-mineral sap transport through TEs is enabled by the spatially and temporally controlled lignification of the SCW. Lignification occurs post-mortem by the supply of monomers and ROS from neighbouring XP cells and is restricted to specific deposition domains, defined by the pre-mortem sequestration of phenoloxidases.

La palourde japonaise, Venerupis philippinarum, a été introduite en France au début des années 70 à des fins aquacoles. Depuis 1987, d'importants épisodes de mortalité, causés par la bactérie pathogène Vibrio tapetis, touchent cette espèce le long des côtes françaises et européennes. Cette vibriose, appelée Maladie de l'Anneau Brun (MAB), est considérée comme une maladie d'eau froide. L'interaction tripartite entre V. philippinarum, V. tapetis et l'environnement a été explorée à travers le rôle du système enzymatique des phénoloxydases (POs) dans le but de mieux comprendre la réponse immunitaire de la palourde japonaise, la virulence de l'agent pathogène mais aussi l'impact de l'environnement et plus particulièrement de la température. Les POs sont des oxydoréductases impliquées dans la synthèse de la mélanine et de ses dérivés mais aussi dans les processus de reconnaissance du non-soi et d'encapsulation chez les invertébrés. Dans un premier temps, l'activité PO du sérum d'hémolymphe a été caractérisée d'un point de vue biochimique comme étant majoritairement de type laccase ; une activité minoritaire de type tyrosinase a également été identifiée. Des infections de palourdes par trois souches de V. tapetis, à différentes températures, ont mis en évidence une modulation de la réponse du système PO en fonction du temps et du compartiment étudiés. De façon générale, l'infection bactérienne s'est traduite par une augmentation de l'activité PO. Toutefois, le niveau basal d'activité PO est variable d'une population à une autre et cette variabilité semble traduire une susceptibilité différente à la MAB. L'augmentation de la température de 15 à 22°C a entraîné une augmentation des capacités immunitaires de la palourde japonaise. La températurea également eu un impact sur la pathogénicité de V. tapetis et ce, de façon différentielle selon les souches. L'inhibition de l'activité PO observée in vitro en présence de produits extracellulaires bactériens souligne la complexité de l'interaction entre V. philippinarum et V. tapetis. Ainsi, le suivi de l'activité PO constitue un biomarqueur pertinent des capacités immunitaires des invertébrés marins dans l'interaction tripartite hôte-pathogène-environnement.

Microbial enzymes implicated in plant cell hydrolysis may have several potential aplications such as biomass degradation biocatalysts or with biofuel production. Bark beetles establish symbiosis with several microbial strains which play different roles benifitting the beetle, as the production of hydrolytic enzymes to degrade the ingested wood, the protection against mirobial antagonist or the detoxification of the environment. Fungal symbionts have been traditionally the best studied, but several recent research with bacterial symbionts of several bark beetle species show that bacterial also display important functions for the host. In this study, the bacterial communities of the bark beetle species Cryphalus piceae and Pithophtorus pithophtorus, collected in the Czech Republic from pine and fir trees, respectively, were isolated and 55 out of 89 samples were identified by 16S rRNA gene amplification and sequencing. Members of the genera Erwinia, Pantoea, Curtobacterium, Yersinia, Pseudomonas and Staphylococcus were detected. The isolates were object of study for their possible biotechnological potential in (ligno)cellulose materials degradation by screening several enzymes implicated in plant cell hydrolysis, as cellulases, xylanases, amylases, laccases, as well as their capability for colorant...