Les écosystèmes tropicaux, en particulier les écosystèmes amazoniens, sont connus pour abriter une importante diversité d'organismes, terrestres ou aquatiques. Cependant, les causes et les processus responsables de cette grande diversité dans les assemblages de poissons d'eau douce restent encore peu connus, et leur identification est un fort enjeu dans l'évaluation future des impacts des perturbations humaines qui sont grandissantes dans ces milieux. Nous avons étudié les processus qui façonnent la diversité et la structure des assemblages de poissons d'eau douce de Guyane dans les petits cours d'eau et les fleuves non impactés par les activités humaines. La diversité au sein des assemblages de petits cours d'eau augmente le long du gradient amont/aval et dans les milieux où l'habitat est plus diversifié. L'identité des espèces change le long de ce gradient, définissant des zones le long du cours d'eau. Les relations spatiales entre les assemblages et leur isolement ont aussi un fort effet sur les assemblages. En incorporant les informations sur les traits des espèces (diversité fonctionnelle) et leur relation de parenté (diversité phylogénétique), nous avons montré que la diversité au sein des assemblages n'était pas influencée par l'environnement ou les interactions entre les espèces. Nous avons aussi confirmé le fort effet de la limite à la dispersion entre les assemblages, en lien avec l'histoire passée des bassins versants, dans les petits cours d'eau et dans les fleuves. De telles études sur les processus structurant les assemblages nécessitent l'acquisition de données biologiques plus complètes, et donc le développement d'une nouvelle méthode d'échantillonnage qui soit exhaustive et non invasive. Pour cela, nous avons testé le metabarcoding environnemental (l'identification moléculaire des espèces présentes à partir d'un échantillon d'eau). Cette méthode donne des résultats complémentaires aux pêches traditionnelles et nécessite encore un travail de développement et des tests supplémentaires pour améliorer son efficacité et permettre son utilisation pour identifier les processus structurant les assemblages. Ces travaux, aussi bien pratiques que théoriques, sont nécessaires au développement d'un meilleur cadre conceptuel sur la structure des assemblages de poissons tropicaux, et dans la construction d'indicateurs d'impacts d'activités humaines sur les écosystèmes. / Tropical ecosystems, especially Amazonian ecosystems, host a great diversity of terrestrial and aquatic organisms. However, the causes and the processes behind this high diversity for freshwater fish assemblages are little known, but their identification will be an asset in the assessment of anthropogenic impacts that are increasing in these regions. We studied the processes that shape the diversity and the structure of freshwater fish assemblages of non-impacted streams and rivers located in French Guiana. Within-assemblage diversity increased along an upstream/downstream gradient and was higher in sites where the habitat was diversified. Species identity changed along this gradient, which created zones along the stream. Spatial relationships between assemblages and their isolation also greatly impacted species assemblages. Using information about species traits (functional diversity) and their phylogenetic relationships (phylogenetic diversity), we showed that within-assemblage diversity was not influenced by the environment or by species interactions. We also confirmed that dispersal limitation, linked with the past history of drainage basins, had a strong effect on assemblage structure in both streams and in rivers. Future investigations on the processes structuring fish assemblages will need to acquire more exhaustive biological data, and therefore to develop an efficient, and non-destructive sampling method. To this aim, we evaluated the efficiency of environmental metabarcoding applied to aquatic assemblages (the molecular identification of species present from a water sample) and compared it to traditional sampling methods. Currently, metabarcoding gives complementary information to traditional sampling. It thus needs developments and further tests to increase its efficiency and allow its use for assembly processes studies. Pursuing the formalization of a conceptual framework to investigate assembly rules together with the development of an efficient fish sampling protocol are now needed to better understand the structure of tropical fish assemblages. Those theoretical and practical developments will contribute to better evaluate anthropogenic disturbances on aquatic ecosystems.

La lignée verte (i.e. les plantes) est représentée dans l'océan par les algues appartenant aux Chlorophyta. Celles-ci contribuent en moyenne à 25% des séquences photosynthétiques (Dinoflagellés exclus) retrouvées dans les inventaires moléculaires pan-océanique. Bien que les Chlorophyta jouent un rôle important dans l'écologie de l'océan et nous permettent de comprendre l'histoire évolutive des plantes terrestres, leur diversité et leur distribution dans les eaux marines du globe est peu documentée. Après avoir optimisé une base de données de séquences de références ARNr 18S, j'ai procédé à l'analyse de données de métabarcodes produits par Ocean Sampling Day (OSD), qui a fourni des données utilisant 2 régions hypervariables du gène du 18S appelées V4 et le V9. La comparaison des images de diversité produites par le V4 et le V9 a illustré l'influence de la base de référence sur la diversité. Ensuite, l'analyse de l'ensemble des V4 a permis d'étudier la distribution des Chlorophyta dans l'océan mondial. De plus, la vérification de l'assignation automatique des OTUs par reconstruction phylogénétique grâce à la base de référence produite en début de thèse, a permis de confirmer l'existence de nouvelles lignées de prasinophytes et de confirmer que la classe des Mamiellophyceae dominait les eaux côtières, alors que les clades VII et IX des prasinophytes dominaient les eaux océaniques oligotrophiques. Ces analyses ont aussi permis de montrer l'écart entre la diversité environnementale et celle dans la base de référence, en particulier pour les genres Ostreococcus et Micromonas (Mamiellophyceae). / The green lineage that dominates on land is represented by Chlorophyta which account in average for 25% of photosynthetic sequences (Dinoflagellates excluded) in global marine molecular inventories. Although Chlorophyta are major keys for ecological understanding of the ocean, as well as the evolutionary story understanding of land plants, their diversity and distribution in marine waters has been understudied. This thesis aims at investigating the environmental diversity of marine Chlorophyta and describing their distributions based on available large scale metabarcoding datasets. First, a reference database of publicly available 18S rRNA sequences of Chlorophyta was assembled and critically curated. Next, the Ocean Sampling Day (OSD) 18S metabarcode datasets were analysed. Chlorophyta diversity was compared for a limited sample set based on two regions of the 18S rRNA: the V4 and V9 regions. Then, Chlorophyta distribution was studied using the full OSD V4 dataset. Careful taxonomic investigations using both automatic and hand checked assignation of OTUs using alignments and phylogenies allowed to confirm the existence of new environmental prasinophytes clades and to confirme, that the Mamiellophyceae were the major group in coastal waters, while prasinophytes Clade VII and IX were dominating the oceanic oligotrophic stations. Comparing V4 and V9 regions illustrated the influence of the reference database on diversity. Moreover, the taxonomic investigation highlighted the diversity gaps between reference databases and environmental datasets. This work emphasizes the neglected importance of Chlorophyta in marine waters and provides some suggestions for future research.

Les Collodaires (Radiolaires) sont des eucaryotes unicellulaires (protistes) marins appartenant au super-groupe des Rhizaria. Tandis que certains sont caractérisés par un mode de vie colonial, d’autres sont observés sous la forme de larges organismes solitaires. Les Collodaires sont des protistes hétérotrophes, prédateurs de plancton, mais également hôtes systématiques de micro-algues photosynthétiques intracellulaires. Les récentes analyses de leur diversité moléculaire dans l’environnement ont démontré leur importante contribution aux communautés planctoniques ainsi que leur distribution globale dans l’océan mondial. Cependant, nos connaissances sur leur diversité, biogéographie et écologie restent paradoxalement parcellaires. La première partie de cette thèse a été dédiée à des études morphologiques détaillées (en microscopie électronique et optique) et combinées à une phylogénie moléculaire élaborée en séquençant les sous-unités 18S et 28S de l’ADN ribosomal pour 75 spécimens, coloniaux ou solitaires. Ce travail a abouti à la réévaluation de la classification des Collodaires et à l’élaboration d’une base de référence morpho-moléculaire robuste. Par la suite, ce cadre de référence morpho-moléculaire a permis d’explorer la biodiversité des Collodaires grâce à une approche de metabarcoding appliquée à une série d’échantillons collectés dans l’océan mondial pendant l’Expédition Tara Océans. La distribution cosmopolite à la surface des océans des différents taxons qui composent les Collodaires, a révélé une diversité plus importante dans les vastes régions océaniques intertropicales et oligotrophiques. Les Collosphaeridae ont été principalement observés en pleine mer alors que les Sphaerozoidae formaient la famille dominante dans les régions côtières, où la biodiversité des Collodaires était plus faible. Les Collophidiidae, formellement décrits au cours de thèse, ont rarement été rencontrés dans les zones photiques, quelque que soit la latitude, suggérant ainsi qu’ils occupent une niche écologique particulière. Enfin, j’ai également employé la technologie d’imagerie in situ Underwater Vision Profiler (UVP5) afin d’explorer de façon quantitative les abondances et biomasses des Collodaires et des Rhizaria, à travers l’océan mondial. Cette approche a révélé que les Rhizaria forment un composant majeur du méso- et macro-plancton, et représentent jusqu’à 4,5% de la biomasse globale des 200 premiers mètres de l’océan mondial. Plus particulièrement dans les 100 premiers mètres, les Collodaires constituent le groupe le plus important des Rhizaria et leur distribution suggère que la photosymbiose pourrait influencer leur succès dans les régions oligotrophiques où ils sont particulièrement abondants. Au-delà d’améliorer notre compréhension de la diversité, la biogéographie et l’écologie des Collodaires dans l’océan mondial, ce travail de thèse souligne la pertinence de combiner et d’utiliser des approches alternatives d’échantillonnage et d’analyses tel que le séquençage haut-débit et l’imagerie in situ dans l’étude des protistes marins dans leur environnement. / Collodaria (Radiolaria) are unicellular marine eukaryotes (protists) belonging to the super-group Rhizaria. Collodarian species contribute to planktonic communities as large solitary cells or can form large gelatinous colonies. They are heterotrophic organisms feeding on other plankton, which also systematically harbour intracellular symbiotic microalgae. Recent environmental molecular diversity surveys demonstrated their important contribution to planktonic communities and their worldwide occurrence in the global ocean. However, knowledge on their diversity, biogeography and ecology is paradoxically very poor. In the first part of this thesis I performed detailed morphological analyses (electron and optical microscopy) combined with a molecular phylogeny based on the 18S and 28S rRNA genes, sequencing for a total of 75 distinct colonial and solitary specimens. Ultimately, this work led to the revision of the Collodaria classification and to the construction of a robust morpho-molecular reference database. Then, this morpho-molecular framework allowed the exploration of Collodaria biodiversity through a metabarcoding approach across samples collected in the global ocean during the Tara Ocean expedition. The cosmopolitan distribution of the different collodarian taxa in the surface oceans revealed a higher biodiversity in the vast oligotrophic inter-tropical open oceans. Collosphaeridae were predominantly found in the open oceans while the Sphaerozoidae were the dominant family in the less diverse coastal regions. The newly defined Collophidiidae were rarely encountered in the photic zones at all latitudes, suggesting that they inhabit a different ecological niche. Finally, I also used the in situ imaging system Underwater Vision Profiler (UVP5) to quantitatively explore the abundances and biomasses of collodarian and rhizarian in the global ocean. This approach revealed that the Rhizaria were a major component of the meso- and macro-plankton, constituting up to 4.5% of the global carbon standing stock in the upper 200 m of the world oceans. More specifically, Collodaria were the most important rhizarian groups in the first 100 m of the oceans, and their distribution suggested that photosymbiosis might be an important factor explaining their success in oligotrophic regions where they are particularly abundant. Besides the improvement of our knowledge on the diversity, biogeography and ecology of Collodaria in the global ocean, this thesis highlights the relevance to combine and/or use alternative sampling and analytical procedures such as high-throughput sequencing and in situ imaging technologies to study marine protists in their environment.

Sorghum crop has become attractive to breeders due to its drought tolerance, and many uses including a human food source, animal feed, industrial fibre and bioenergy crop. Sorghum, like any other plant, is a host to a variety of microbes that can have neutral, negative or positive effects on the plant. While the majority of microorganisms are beneficial, pathogens colonize plant tissues and overwhelm its defence mechanisms. This colonization is a direct threat to the sorghum productivity. The development of microbiome-based approaches for sustainable crop productivity and yield is hindered by a lack of understanding of the main biotic factors affecting the crop microbiome. Metabarcoding has proven to be a valuable tool which has been widely used for characterizing the microbial diversity and composition of different environments and has been utilized in many research endeavours. This study analyses the relationship between the microbiota and their response to natural pathogen infection in sorghum disease groups (R, MR, S and HS) and identifies the most dominant pathogen in the highly susceptible disease group. The study also, assesses the spore viability through the use of the automated cell counter and confirms Fusarium graminearum (dominant pathogen linked to the HS disease group) through sequencing of the marker genes, to subsequently characterize pathways likely to be involved in pathogen infection resistance. To achieve the objectives, a combination of 16S rRNA (V3/V4 regions) and ITS (ITS1/ITS4) of the internal transcribed spacer regions were amplified and sequenced using NGS technologies to study the microbiota in response to natural infection. Additionally, comparative transcriptional analysis of sorghum RILs in response to Fusarium graminearum infection was conducted through RNA-Seq. Upon natural infection, the foliar symptoms assessment of the RILs was conducted and four disease groups; resistant (R), moderately resistant (MR), susceptible (S) and highly susceptible (HS) were designated. The results of the present metabarcoding study indicate that resistant sorghum leaves (R group) supported a large diversity of fungal and bacterial microbes. The genera Methylorubrum, Enterobacter and Sphingomonas with reported plant growth promoting traits were more abundant and highly enriched in the R and MR group, with members of the latter genus significantly enriched in the R group. The resistant fungal group had a majority of OTUs showing similarity to well-known plant growth-promoting fungal genus including Papiliotrema (Tremellaceae family), which are known biocontrol agents. The yeast Hannaella was also highly linked with the resistant plants. Some Hannaella species are known to produce indole acetic acid (IAA) for promoting plant growth. Metabarcoding was also used to assess the major potential disease-causing taxa associated with the highly diseased group. It identified fungal pathogenic species, that have not previously been identified as pathogens of sorghum such as Ascochyta paspali and Ustilago kamerunensis (which are known pathogenic fungi of grass species) and were associated with the susceptible disease groups (S and HS). These analyses revealed the potential sorghum fungal pathogen Epicoccum sorghinum, and was highly linked with the S disease group. It further expanded the identification of a reportedly economically importance species causing sorghum related diseases Fusarium graminearum (anamorph Gibberella zeae). This species has also been identified in this study to be highly associated with the RILs showing major disease symptoms. Fusarium graminearum a significant pathogen in winter cereals and maize has been associated with stalk rot of sorghum and sorghum grain mould. The presence of Fusarium graminearum in sorghum can be a toxicological risk, since this species has the potential to produce mycotoxins. It was further shown that natural pathogen infection results in distinct foliar microbial communities in sorghum RILs. The co-occurrence taxa represented by Tremellomycetes and Dothiomycetes fungal classes and Bacillaceae and Sphingomonadaceae bacterial family had more central roles in the network. The modules which are located centrally on the network have been expected to play important ‘topological roles’ in interconnecting pairs of other fungal and bacterial taxa in the symbiont–symbiont co-occurrence network. These taxa having a central role, are considered to be keystone microbes, and have been suggested to be drivers of microbiome structure and functioning. The results of bacterial and fungal community composition, community co-occurrences further suggested the importance of keystone taxa which may disproportionately shape the structure of foliar microbiomes. The foliar disease symptom assessments revealed that sorghum RIL 131 was highly diseased and RIL 103 did not show any visible disease symptoms and were subsequently used for transcriptomic analysis. Gene expression patterns were studied between the identified RIL that did not show visible symptoms (resistant RIL no 103) and the RIL that showed major disease symptoms (susceptible RIL no 131). Fusarium graminearum the dominant potential pathogen found in this study to be associated with the highly susceptible plants was used to inoculate RILs at seedling stage in a greenhouse and samples were collected in triplicates at 24 hours post infection (hpi), 48 hpi, 7 days post infection (dpi) and 14 dpi. Prior to that, ITS and UBC genes confirmed the identity of Fusarium graminearum, and the automated haemocytometer confirmed the cell/spore viability. Using RNA-Seq analysis it was shown that the resistant RIL had defence related pathways from early response (24- 48 hpi) to late response (7-14 dpi). And the more the infection progressed, the more the defence related genes were up-regulated in terms of fragments per kilobase of exon model per million reads mapped (FPKM) and False Discovery Rate (FDR ≤ 0.05) values. Transcriptome time series expression profiling was used to characterize the plant response to Fusarium graminearum with the Dirichlet Process Gaussian Process mixture model software (DPGP) in susceptible and resistant RILs. The susceptible RIL (number 131) transcriptional response upon Fusarium graminearum infection presented differences of the closely related clustered expression profiles across all timepoints in both RILs. Group 2 exclusively clustered the genes encoding the sesquiterpene metabolism pathway, which is one of the major physiological change occurring in response to fungal infection and has been previously reported to produce the mycotoxins associated with Fusarium head blight (FHB) of cereals. This pathway presented an increase from the initial infection phase to the late infection phase in group 4, the genes encoding starch sucrose, metabolism and cyanoamino acid pathways presented a pattern that had a sharp decline from 48 hpi -14 dpi (at a later stage of infection). This could suggest that, as the time progresses in the susceptible RIL the pathways which are important in plant defence declines at a late infection stage. Group 3 presented a pattern increase of the 5-lipoxygenase (LOX 5) gene expressed from 48 hpi-14 dpi timepoints. The loss and silencing of LOX5 function have in the past described to be linked with enhanced disease resistance. In this study the LOX5 was expressed and this could suggest that LOX5 might have a function as a susceptibility factor in disease caused by Fusarium graminearum in sorghum RILs. CBL-interacting protein kinase 6 (CIPK6) gene was also associated with this group. This gene has been associated with negative regulation of immune response to Pseudomonas syringae in Arabidopsis as plants overexpressing CIPK6 were more susceptible to Pseudomonas syringae. Transcriptional response of a resistant RIL (number 103) to infection with Fusarium graminearum presented an increase in genes encoding metabolic and biosynthesis of metabolites pathways in group 1 and group 4 at early infection phase and a sharp decline in the late infection phase. An increase in the genes encoding pathways in earlier infection state could suggest the establishment of a beneficial energy balance for defence. Additionally, genes encoding phenylpropanoid (PAL), galactose and glycolysis pathway were amongst the genes increased at early stages of infection in group 1. Sugar can play a significant role in resistance to fungal pathogens through phenylpropanoid metabolism stimulation, and previous studies showed that the phenylpropanoid pathway could play a role in resistance of wheat to Fusarium graminearum and deoxynivalenol. Overall, this study represents a first step in understanding the molecular mechanisms involved in resistance to Fusarium graminearum. This analysis has also identified the reported beneficial microbes and defence related genes and pathways. Together, the current findings suggest that different ‘resident’ consortia found in naturally infected and uninfected sorghum plants may be viable biocontrol and plant-growth promoting targets. Cultivation studies may shed light on the nature of the putative symbiotic relationships between bacteria and fungi. These results have consequences for crop breeding, and the analysis of microbial diversity and community composition can be useful biomarkers for assessing disease status in plants. The transcriptome and metabarcoding data generated will help guide further research to develop novel strategies for management of disease in sorghum RILs through the integrative approach considering both beneficial microbes and defence related genes. This provides the baseline information and will positively impact in the development of Fusarium graminearum resistant genotypes in future through the integration/incorporation of beneficial microorganisms (bacteria and fungi) and resistant genes in breeding strategies. / Life and Consumer Sciences / D. Phil. (Life Sciences)

Analysis of soil-derived DNA has been shown to minimize problems seen during traditional vegetation surveys, e.g. by matching the eDNA to a reference database for taxonomic identification rather than relying solely on taxonomic expertise. However, it has been debated to what extent and how accurately eDNA acts as a proxy for biodiversity. The reliability on eDNA and the awareness on influencing factors is also important for palaeoenvironmental reconstructions where above-ground vegetation cannot be used. This study aimed to investigate how well modern soil-derived eDNA reflects the contemporary vascular vegetation communities in a subarctic environment, and if the efficiency of the taxonomic identification differed between spatial scales. Near-surface soil samples at altitudinal gradients along numerous transects were collected in combination with vegetation surveys. The eDNA was amplified through metabarcoding using the P6 loop region of the chloroplast trnL intron, followed by a high-throughput sequencing. No difference in the number of identified taxa between eDNA and vegetation survey was seen at landscape scale. In contrast, the number of identified taxa was consistently higher in the vegetation survey at smaller spatial scales. The efficiency of identified taxa per scale remained stable for the vegetation survey, whereas for eDNA, a decreasing trend was seen. This study highlights the variations on taxa identification between both methods and which factors might cause it. Combining the methods allows for a more precise modern biodiversity estimation, as well as to minimize wrongful conclusions. This allows for a more accurate palaeoenvironmental reconstructions, which in turn can improve future species conservation decisions.

Despite over four decades of research on deep-sea hydrothermal vent ecosystems, major gaps remain in our understanding of these systems. Knowledge of microeukaryote diversity, abundance, and involvement in ecosystem function lags far behind that of prokaryotes, and contributions of the non-endosymbiotic microbiome in faunal assemblages to ecosystem processes and overall hydrothermal vent microbial diversity are not known. This research addresses these gaps using high-throughput sequencing of 16S/18S rRNA genes and metagenomes from vent and surrounding non-vent habitats encompassing diffuse hydrothermal fluids, plumes, deep seawater, and microbes in assemblages of the foundation tubeworm species Ridgeia piscesae. Co-occurrence/covariance is a central method used, first, between prokaryotes with known extreme habitat preferences and microeukaryotes to infer potential endemism in the latter, and then between microbes and fauna in R. piscesae assemblages to infer interspecies interactions. Microeukaryote distribution and abundance suggest potential vent endemic microeukarya are infrequently encountered, potentially in low abundance, and belong to novel lineages of Rhizaria and Stramenopila. Potential endemism is inferred for relatives of known apusomonads, excavates, and some clades of Syndiniales. R. piscesae assemblages are shown to be hotspots of microbial taxonomic richness and exhibit a robust temperature-driven distinction in assemblage composition above and below ~25°C spanning micro, meio and macro size classes and microbial domains (Bacteria, Archaea, and micro-Eukarya). Likely interacting faunal and microbial taxa among R. piscesae assemblages are identified as ‘core communities’, which included eight macro- and meiofaunal taxa and members of the Bacteroidetes and Epsilonbacteraeota in highT communities (>25°C) and more meiofaunal species in addition to Alpha- and Gammaproteobacteria, and Actinobacteria in lowT communities (<25°C). Core communities were used to guide metagenomic investigations of microbial functional potential. Exploratory metagenomic analysis required development of new methods to deal with compositional data. ‘Enrichment leanings’ were developed to prioritize in-depth functional comparisons between sample types, which revealed clades within core community microbes with differing functional potential between highT and lowT assemblages and between assemblages and fluids. The balance of autotrophy-heterotrophy genes and patterns of genes for different carbon, nitrogen, and sulfur-cycling processes were tested as potential metrics of community-level function but did not distinguish assemblages by highT/lowT designations. This research brings us closer to understanding hydrothermal vent ecosystem function and suggests sizeable continued discovery potential. / Graduate / 2022-06-08

Travail réalisé en cotutelle avec l'université du Littoral côte d'opale, France / Afin de réduire l’usage des fertilisants chimiques et leurs empreintes environnementales, la production de bioinoculants à base de bactéries rhizosphériques solubilisatrices de Phosphore (PSB) est une biotechnologie prometteuse. Lorsque celle-ci est associée à l’apport de phosphate de roche (RP), elle pourrait constituer une alternative écoresponsable d’amélioration de la fertilité des sols rhizosphériques, notamment à travers l’augmentation du phosphore (P) biodisponible. Pour répondre à cet enjeu environnemental majeur, il est indispensable d’étudier la biodiversité microbienne des sols en interaction avec le RP. Ainsi, ce travail de thèse a pour objectif de caractériser, d’un point de vue taxonomique, les communautés bactériennes et fongiques associées aux compartiments racinaires et rhizosphériques de plantes natives d’environnements singuliers, peu explorés, que sont d’anciens sites miniers riches en RP. Le séquençage Illumina MiSeq d’amplicons de gènes ribosomiques des différents groupes microbiens et l’assignation taxonomique des séquences regroupées en ASV, nous ont permis de mettre en évidence des valeurs de richesse et de diversité microbiennes comparables malgré la différence des teneurs en RP et en P des sols étudiés. De même, une grande similarité entre les profils des communautés a été identifiée à haut niveau taxonomique, avec une prévalence marquée des phyla Actinobacteriota et Ascomycota, et de l’ordre Glomerales pour les communautés de champignons mycorhiziens arbusculaires (CMA). À un niveau taxonomique plus bas, un core microbien de 26 ASV mycorhiziennes, persistantes à travers le gradient de RP et de P, a également été mis en évidence. Toutefois, l’analyse statistique des données environnementales nous a permis d’identifier des ASV indicatrices des habitats enrichis versus non enrichis en RP, pour chaque communauté microbienne étudiée. La synthèse de ces résultats originaux montre une influence mineure des teneurs en RP et en P sur le profil des communautés microbiennes des sites étudiés et questionne la signification écologique de la prévalence de certains taxa en lien avec l’histoire minière du site, sa restauration écologique et sa résilience, ainsi que les stratégies de vie et les attributs fonctionnels des taxa dominants. Par ailleurs, une collection de PSB a été réalisée à partir d’isolats racinaires issus des sites miniers étudiés, ou à partir d’isolats issus de spores et d’hyphes mycorhiziens piégés au contact d’apports en RP déposés dans la rhizosphère de plantes d’intérêt agronomique. Ces PSB racinaires ou hyphosphériques, testés par ailleurs pour plusieurs traits additionnels promoteurs de la croissance des plantes, ont été assignés à trois phyla : Proteobacteria, Firmicutes et, dans une moindre mesure, Actinobacteria ; Bacillus et Pseudomonas étaient les genres prédominants. Enfin, à partir de ces isolats, deux consortia bactériens avec différent traits promoteurs de la croissance des plantes (PGP), ont été constitués et testés seuls ou en combinaison avec l’apport de RP et/ou l’ajout d’un inoculum mycorhizien à base de Rhizophagus irregularis, sur la croissance de la tomate, espèce d’intérêt agronomique. Une augmentation du taux de germination, un accroissement de la hauteur des jeunes plantules et des modifications au niveau du système racinaire des plants adultes, en particulier des variations de la densité du tissu racinaire, ont été observés en réponse à l’inoculation simple par les consortia ou à leur interaction double ou tripartite avec le RP et/ou le CMA. Le profil taxonomique du microbiote natif avant et après apport des inoculants et du RP a été caractérisé. En conclusion, l’ensemble de nos travaux s’appuyant sur l’analyse des données environnementales, combinées aux données de laboratoire sur l’inoculation de plantes par des isolats bactériens, notamment d’origine minière, soulignent l’intérêt mais aussi la complexité de l’élaboration d’inoculants microbiens promoteurs de la croissance des plantes, solubilisateurs de P. / In order to reduce the use of chemical fertilizers and their environmental footprint, the production of bioinoculants based on phosphorus solubilizing rhizospheric bacteria (PSB) is a promising biotechnology. When combined with rock phosphate (RP), it could be an eco-responsible alternative to improve rhizospheric soil fertility, especially by increasing bioavailable phosphorus (P). To address this major environmental issue, studying soil microbial biodiversity and its interactions with RP is required. Thus, the objective of this thesis is to characterize, from a taxonomic point of view, the bacterial and fungal communities associated with the root and rhizospheric compartments of native plants in singular, little explored environments, located in former mining sites rich in RP. Illumina MiSeq sequencing of ribosomal gene amplicons of the different microbial groups, then taxonomic assignment of the sequences grouped in ASV, allowed us to highlight comparable microbial richness and diversity indexes of soils despite their differences in RP and P contents. As well, a high degree of similarity between the community profiles was identified at a high taxonomic level, with a marked prevalence of the phyla Actinobacteriota and Ascomycota, and of the order Glomerales for the arbuscular mycorrhizal fungi (AMF) community. At a lower taxonomic level, a microbial core of 26 mycorrhizal ASV, persistent across the RP and P gradient, was also documented. However, statistical analysis of the environmental data allowed us to identify indicator ASV for PR-enriched versus non-rich habitats for each microbial community studied. All together, these original results contributed to rule on a minor influence of RP and P levels on the microbial community’s profiles across the studied sites, and raised the questions of the ecological significance of the prevalence of certain taxa regarding the site’s mining history, its ecological restoration, as well as the life strategies and functional attributes of the dominant taxa. In addition, a collection of PSB was made from mine site root isolates, or from isolates associated with spores and mycorrhizal hyphae trapped in contact with PR inputs deposited in the rhizosphere of agronomic plants. These root or hyphospheric PSB, tested elsewhere for several additional plant growth-promoting traits, were assigned to three phyla: Proteobacteria, Firmicutes, and, to a lesser extent, Actinobacteria; Bacillus and Pseudomonas were the predominant genera. Finally, from these isolates, two bacterial consortia with different plant growth promoting (PGP) traits were constituted and tested, alone or in combination with RP inputs and/or a mycorrhizal inoculum based on Rhizophagus irregularis, on the growth of tomato, a species with an agronomic interest. An increase in germination rate or height of young seedlings, and changes in the root system of adult plants, particularly changes in root tissue density, were observed in response to single inoculation by the consortia or to their dual or tripartite interaction with RP and/or AMF. The taxonomic profile of the native microbiota before and after inoculants and RP input was characterized. In conclusion, our work based on the analysis of environmental data, combined with laboratory data from plant inoculation experiments by bacterial isolates, especially of mining origin, underlines the interest but also the complexity of the development of microbial inoculants promoting plant growth, solubilizers of P.

The global aquaculture industry is expanding to support increased demand in global markets and supplement traditional fisheries. The rapid increase of aquaculture production relies on introducing and using species outside their native range, posing significant regional environmental and socio-economic risks. Non-native species are selected based on transferable large-scale production protocols, fast growth, and existing market demand. Aquaculture is an important sector within Saudi Arabia’s Vision 2030 (anticipated >400% production increase by 2030). Barramundi (Lates calcarifer) was introduced into the Red Sea for open sea-cage farming in 2008, with large-scale production beginning in 2014. Regionally, there are numerous anecdotal reports of escape events of this species, including an incident in 2015 involving the release of ~300,000 fish. Since then, local fishers have caught up to 15 wild individuals per month in Al Lith, central Saudi Arabia. This study presents the first biological information on L. calcarifer aquaculture escapees in the Saudi Arabian Red Sea. Wild-caught L. calcarifer (n=5) were collected from a local fish landing and measured up to 10 kg, eight years of age, were sexually mature, and consumed benthic and coral reef fishes. In contrast, individuals in aquaculture facilities are harvested at < 1 kg and < 400 mm. In their native range, L. calcarifer can reach up to 200 cm and 60 kg. To assist in detecting and monitoring escapees within the Red Sea, I designed and validated a species-specific SYBR-based environmental DNA quantitative PCR assay targeting a 16S mitochondrial region of L. calcarifer rRNA (Barramundi_16S assay). Preliminary results, using DNA metabarcoding and environmental (seawater) samples, detected L. calcarifer near active/historical aquaculture farms and north/south of areas where this species has not been reported previously in the Red Sea (10–250 km from aquaculture facilities). In the future, the Barramundi_16S assay can screen additional eDNA samples (n=250) collected for this study to delineate the geographic range of barramundi in the Red Sea. Cumulatively, this study highlights the need to consider the ecological impacts of aquaculture escapees and provides managers and industry with valuable information and a novel molecular monitoring tool for detecting aquaculture escapees.

Oomycetes (Stramenopiles, protists) are among the most severe plant pathogens, comprising species with a high economic and ecologic impact on forest ecosystems. Their diversity and community structures are well studied in terrestrial habitats, but tree canopies as huge and diverse habitats have been widely neglected. A recent study highlighted distinct oomycete communities in the canopy stratum compared to the ground region of three temperate deciduous trees (Quercus robur, Tilia cordata, Fraxinus excelsior). While the communities from the two strata were distinct when taking oomycete abundances into account, they were rather similar when only OTU presence/absence was considered. It remains, however, unknown if this homogeneity in the OTU presence also leads to a functional homogenisation among microhabitats within the two strata ground and canopy. In this study, we supplemented functional traits to oomycete communities in the tree microhabitats, which were determined over a time period of 2 years with a metabarcoding approach. Our results showed that even though most oomycetes occurred in all microhabitats, a strong discrepancy between the strata and correspondingly the distribution of oomycete lifestyles could be observed. This pattern was constant over several seasons. Obligate biotrophic species, exclusively feeding on living host tissue, dominated the canopy region, implying tree canopies to be a hitherto neglected reservoir for parasitic protists. OTUs assigned to the genus Hyaloperonospora—parasites highly specialised on hosts that were not sampled—could be determined in high abundances in the canopy and the surrounding air, challenging the strict host dependencies ruled for some oomycetes. Our findings further contribute to the understanding of oomycete ecosystem functioning in forest ecosystems

No description available.