Empresa Brasileira de Pesquisa Agropecuária / The knowledge of the level of association that occurs between diazotrophic bacteria and fungi arbusculares mycorrhizae (AMF) in grassland ecosystems may be important for the understanding of the changes caused by the addition of fertilizers containing phosphorus (P) and nitrogen (N), in the production and botanical composition of natural pastures. The objective of this study was to evaluate. (a) the occurrence of three genera of diazotrophic bacterial in the root system under fertilization with N and P, and determine the contribution of N via BNF, and (b) evaluate the behavior of the association between AMF and native grasses. Four most abundant grasses in natural grasslands of the Southern Campos in Rio Grande do Sul , Axonopus affinis, Paspalum notatum, Andropogon lateralis and Aristida laevis were grown in pots of 5 kg, in a greenhouse, two studies being conducted (A e B ). In A, two treatments were applied: 50 mg kg-1 soil P and 100 mg kg-1 of soil N (NP) and a control, being evaluated, the number of diazotrophic bacteria of the genera Azotobacter, Azospirillum and Herbaspirillum, and the contribution of BNF was determined by the technique of natural 15N abundance. In B, the treatments consisted of applying 50 mg kg-1 soil P (P), application of 50 mg kg-1 soil P and 100 mg kg-1 of soil N (NP), and a control, in both treatments mycorrhizal colonization was determined. For A, A. laevis demonstrate to be more dependent on biological N fixation than the other species. The grass P. notatum compared with other species demonstrated to be more efficient to absorb available soil N. The dry matter accumulation in shoots of the native species was higher with the application of NP. In B the mycorrhizal colonization was similar between the control, P and NP to the roots of A. lateralis and A. laevis, thus presenting a greater dependence on the mycorrhizal association. Differently, in A. affinis and P. notatum, the mycorrhizal colonization was lower when subjected to fertilization with P and NP, thus presenting a lower dependence. It is concluded for A that fertilization with N and P reduces diazotrophic colonization, increasing the production of dry matter and N content of the tissue. A. laevis showed the highest contribution of biological nitrogen fixation, since P. notatum showed higher N accumulation in soil. In relation to B, A. laevis and A. lateralis have a higher dependence on the mycorrhizal than A. affinis and P. notatum. / O conhecimento do nível de associação que ocorre entre bactérias diazotróficas e fungos micorrízicos arbusculares (FMA) nos ecossistemas campestres, pode ser importante para a o entendimento das alterações provocadas pela adição de fertilizantes, contendo fósforo (P) e nitrogênio (N), na produção e na composição botânica das pastagens naturais. O objetivo do presente trabalho foi avaliar (a) a ocorrência de três gêneros de bactérias diazotróficas no sistema radicular sob fertilização com N e P, e determinar a contribuição de N via FBN, e (b) avaliar o comportamento da associação entre FMAs e gramíneas nativas. Foram utilizadas quatro gramíneas de maior abundância nas pastagens naturais dos Campos Sulinos no Rio Grande do Sul, Axonopus affinis, Paspalum notatum, Andropogon lateralis e Aristida laevis, cultivadas em casa de vegetação, sendo conduzidos dois estudos (A e B). Em A, foram aplicados dois tratamentos: 50 mg kg-1 de solo de P + 100 mg kg-1 de N solo (NP) e uma testemunha, sendo avaliados, o número de bactérias diazotróficas dos gêneros Azotobacter, Azospirillum e Herbaspirillum, e a contribuição da FBN através da técnica da abundância natural de 15N. Em B, os tratamentos consistiram na aplicação de 50 mg kg-1 de solo de P (P); aplicação de 50 mg kg-1 de solo de P + 100 mg kg-1 de N solo (NP), e uma testemunha, em ambos foi determinada a colonização micorrízica. A espécie A. laevis demonstrou ser mais dependente da fixação biológica de N que as demais espécies. A espécie P. notatum em comparação as demais espécies, demostrou ser mais hábil em absorver o N disponível no solo. A matéria seca acumulada da parte aérea das espécies nativas foi maior com a aplicação de NP. A colonização micorrízica foi semelhante entre a testemunha, P e NP para as raízes de A. lateralis e A. laevis, apresentando assim uma maior dependência da associação micorrízica. Diferentemente, nas espécies A. affinis e P. notatum, a colonização micorrízica foi menor quando submetidas a adubação com P e NP, apresentando assim uma menor dependência. Conclui-se assim, que a adubação com N e P reduz a colonização de bactérias diazotróficas, aumentando a produção de matéria seca e teor de N no tecido. A. laevis apresentou a maior contribuição da fixação biológica de nitrogênio, já P. notatum apresentou maior acúmulo de N do solo. As espécies A. laevis e A. lateralis apresentam maior dependência da micorrização que as espécies A. affinis e P. notatum.

La phytoremédiation, la technique de bioremédiation qui utilise les plantes, est considérée comme une technologie « verte » et efficace pour décontaminer des sols pollués aux hydrocarbures. Les plantes agissent essentiellement à travers la stimulation des microorganismes de la rhizosphère, où l’exsudation racinaire semble être le moteur majeur de l’activité microbienne qui s’y déroule. Beaucoup de gènes responsables de l’adaptation bactérienne, dans le sol contaminé ou dans la rhizosphère, semblent être portés par les plasmides conjugatifs et transférés entre les bactéries. Une meilleure compréhension de ce phénomène pourrait améliorer le développement de techniques de manipulation du microbiome rhizosphérique afin d’accélérer la bioremédiation. Le but de cette recherche est d’étudier le plasmidome, soit le contenu total en plasmides, de la rhizosphère de saules provenant d’un sol contaminé aux hydrocarbures. Des analyses métagénomiques, de données obtenues à partir d’un séquençage Illumina, ont été utilisées pour étudier les fonctions portées par les plasmides. Nos résultats ont indiqué un fort effet de la contamination aux hydrocarbures sur la composition des plasmides. De plus, les plasmides contenaient des gènes impliqués dans de nombreuses voies métaboliques, telles que la biodégradation d’hydrocarbures, la production d’énergie, la transduction du signal, le chimiotactisme, et les métabolismes des sucres, acides aminés et métabolites secondaires. À ce jour, c’est la première étude de métagénomique comparative documentant la diversité des plasmides dans un contexte de phytoremédiation. Ces résultats fournissent de nouvelles connaissances sur le rôle du transfert latéral de gènes dans l’adaptation bactérienne dans la rhizosphère et dans le sol contaminé aux hydrocarbures. / Phytoremediation, a bioremediation technique that uses plants, is considered to be an effective and affordable “green technology” to clean up hydrocarbon contaminated soils. Plants essentially act indirectly through the stimulation of rhizosphere microorganisms. Root exudation is thought to be one of the predominant drivers of microbial communities in the rhizosphere and is therefore a potential key factor behind enhanced hydrocarbon biodegradation. Many of the genes responsible for bacterial adaptation in contaminated soil and the plant rhizosphere are thought to be carried by conjugative plasmids and transferred among bacteria. A better understanding of these phenomena could thus inform the development of techniques to manipulate the rhizospheric microbiome in ways that improve hydrocarbon bioremediation. This research aims to study the plasmidome (the overall plasmid content) in the rhizosphere of willow growing in hydrocarbon contaminated and non-contaminated soils, as compared with unplanted soil. Metagenomic analyses based on Illumina sequencing were used to highlight functions carried by plasmids. Our results indicate a strong effect of hydrocarbon contamination on plasmid composition. Furthermore, plasmids harbored genes involved in several metabolic pathways, such as hydrocarbon biodegradation, energy production, signal transduction, chemotaxis, metabolisms of carbohydrates, amino acids and secondary metabolites. To date, this is the first comparative soil metagenomics documenting the plasmidome diversity in a phytoremediation system. The results provide new knowledge on the role of lateral gene transfer in the bacterial adaptation in rhizosphere and in hydrocarbon contaminated soil.

In dieser Arbeit wurden zunächst die Kolonisationen von drei verschiedenen Pflanzengattungen durch den GFP-markierten Bacillus amyloliquefaciens FZB42 mittels confocaler Lasermikroskopie und Elektronmikroskopie verfolgt. Hier konnte gezeigt werden, dass FZB42 alle ausgewählten Pflanzen besiedeln konnte. Bei Arabidopsis- und Maiskeimlingen wurden die Wurzelhaare und Verbindungen, an denen laterale Wurzeln entstehen, durch FZB42 bevorzugt besiedelt. Weiterhin wurden bei Arabidopsis die Spitzen der Primärwurzeln, und bei Mais die Wurzelkerben bevorzugt besiedelt. Bei Lemna wurden FZB42 Zellansammlungen entlang der Furchen, die zwischen den Epidermiszellen der Wurzel liegen, sowie den intrazellulären Hohlräumen an der Blattunterfläche gefunden. Anschließend wurden die Transkriptome von FZB42, der mit Maiswurzelexudat angezogen wurde, mittels Microarray analysiert. Insgesamt wurden 302 Gene, die 8,2 % des Transkriptoms ausmachen, signifikant durch das Wurzelexudat beeinflusst, wobei die Mehrzahl (260 Gene) hochreguliert wurde. Die induzierten Gene, dessen Funktion bereits bekannt ist, sind hauptsächlich an dem Nährstoffwechsel, Chemotaxis und Beweglichkeit, sowie an der Produktion von Antibiotika beteiligt. Auch wurden die Trankriptome von sieben FZB42-Muatnten durch Microarray analysiert. Diese hatten jeweils eine Deletionen in fünf Sigmafaktor-Genen (sigB, sigD, sigM, sigV,and sigX) und zwei globalen Transkriptionsregulator-Genen (degU und abrB). Die Expression vieler Genen wird durch diese Genprodukte beeinflusst. Mögliche Mechanismen, wie diese Faktoren die bakterielle Reaktion auf Wurzelexsudaten beeinflüssen, wurden vorgeschlagen. Schließlich wurden Northernblott-Untersuchungen an möglichen sRNA-Kandidaten durchgeführt, dessen Expression signifikant durch Wurzelexudate beeinflusst wurde. Dabei konnten 6 von 20 vermeintlichen sRNA-Kandidaten betätigt werden. Dies weist auf eine noch unbekannte Rolle der sRNAs bei der Pflanzen-Mikroben-Wechselwirkung. / In this work colonization of three different plants genera, maize, Arabidopsis, and Lemna, by GFP-labeled Bacillus amyloliquefaciens FZB42 in a gnotobiotic system was firtly studied using confocal laser scanning microscopy and electron microscopy. It was shown that FZB42 is able to colonize all these three plants with a specific pattern. Root hairs and the junctions where lateral roots occurred were a preferred area of FZB42 on both maize and Arabidopsis seedlings. On Arabidopsis, tips of primary roots were another favored site of FZB42; while, on maize, the concavities in root surfaces were preferred. FZB42 cells were also able to colonize Lemna, preferably accumulating along the grooves between epidermis cells on roots and the concaved intercellular space on fronds. Secondly, microarray experiments were performed concerning the transcriptomic response of FZB42 to maize root exudates. A total of 302 genes representing 8.2% of FZB42 transcriptome were significantly altered in transcription by the presence of root exudates, the majority of them (260) were up-regulated in expression. The induced genes with known function were mainly involved in nutrition utilization, chemotaxis and motility, and antibiotic production. The transcriptome of seven FZB42 mutants, defective in five sigma factor genes (sigB, sigD, sigM, sigV, and sigX) and two global transcriptional regulator genes (degU and abrB), were also investigated through microarray experiments. A vast number of genes were indentified to be controlled by the protein factors respectively. Possible mechanisms were proposed of how these protein factors are involved in the response to root exudates. Finally, by northern blot existence of six out of 20 small RNA (sRNA) candidates was identified, which were significantly altered in expression by root exudates. This suggests that sRNA may play a hitherto unrecognized role in plant-microbe interaction.

Bacillus amyloliquefaciens FZB42 ist ein frei lebendes Bakterium, das Pflanzenwurzeln besiedelt und das Pflanzenwachstum durch viele verschiedene Wirkmechanismen anregt. In dieser Arbeit wurden die molekularen Grundlagen dieser positiven Wirkungen, die dieses „Pflanzenwachstum fördernde Rhizobakterium“ (PGPR) auf seine Wirte ausübt, untersucht. Um den gegenseitigen Austausch von B. amyloliquefaciens und seinen Wirtspflanzen zu entschlüsseln, wurden umfangreiche Proteomstudien durchgeführt. Es wurden Referenzkarten der extrazellulären und zytosolischen Proteinfraktionen erstellt. Die größte Anzahl an ausgeschiedenen Proteinen konnte während der stationären Phase beobachtet werden. Die identifizierten extrazellulären Proteine gehören verschiedenen Funktionsklassen an, wobei die prominentesten Klassen am Kohlenhydrat-Abbau und den Transport von Molekülen durch die Zellwand beteiligt sind. Die zytosolischen Extrakte von Kulturen, die in 1C-Medium bzw. Mineralmedium angezogen wurden, und in der zweidimensionalen Gelelektrophorese (2 DE) aufgetrennt wurden, ergaben 461 und 245 verschiedene Protein-Einträge. Die erstellten Referenz-Karten wurden anschließend verwendet, um Proteine und Prozesse, in an der Interaktion mit Pflanzen beteiligt sind, zu identifizieren. Dafür wurden die Bakterien Wurzelexudaten von Mais (Zea mays L.) ausgesetzt. Die Proteine aus zwei Stämmen, denen die globalen Transkriptionsregulatoren (Degu, AbrB) und vier Sigma-Faktoren (SigB, SigM, SigV, und SigX) fehlen, wurden ebenfalls untersucht, um ihre Beteiligung an den bakteriellen Reaktionen auf die Wurzelausscheidungen zu analysieren. Zusammenfassend ist dies die erste Studie, die umfangreiche Proteomdaten von Gram-positiven PGPR präsentiert, wobei gleichzeitig die Veränderung der Expression von extrazellulären und zytoplasmatischen Proteinen, nach Zugabe von Wurzelexudaten, ausgewertet wurde. / Bacillus amyloliquefaciens strain FZB42 is a free-living bacterium that competitively colonizes plant roots and stimulates plant growth by many different modes of action. The molecular basis of singular beneficial effects that this Plant Growth-Promoting Rhizobacteria (PGPR) exert on their hosts have been studied. To decipher the molecular cross-talk of B. amyloliquefaciens and its’ host plants as a whole system, an extensive proteomic approach was performed. Reference maps of the extracellular and cytosolic protein fractions were established. The highest number of secreted proteins was observed during stationary growth phase. Identified extracellular proteins belong to different functional classes, with the most prominent classes involved in carbohydrate degradation and transportation of molecules across the cell wall. Cytosolic extracts obtained from cultures grown in 1C and minimal media subjected to the 2 Dimensional Electrophoresis (2 DE), revealed 461 and 245 different protein entries, respectively. Created reference maps were subsequently used to identify proteins and processes involved in the interaction with plants, prior to exposure of bacteria to maize (Zea mays L.) root exudates. The proteomics of two strains lacking expression of genes coding for global transcriptional regulators (degU, abrB) and four sigma factors (sigB, sigM, sigV, and sigX) were also inves-tigated, in order to analyse their involvement in bacterial responses to root exudates. In summary, this is the first study presenting comprehensive proteomics of Gram-positive PGPR, evaluating at the same time changes in protein expression caused by addition of root exudates at the extracellular and cytosolic level.

Interactions between plant roots and soil microorganisms in the rhizosphere are critical for plant growth. However, understanding of precisely how root exudates influence the diversity and activity of rhizosphere microorganisms is limited. The main objective of this study was to investigate the effect of radiata pine (Pinus radiata) root exudates on rhizosphere soil microbial communities, with an emphasis on the role of low molecular weight organic anions. The study involved the development and validation of new methods for investigating rhizosphere processes in a purpose-built facility. This included development of an in situ sampling technique using an anion exchange membrane strip to collect a range of organic anions exuded from radiata pine roots grown in large-scale rhizotrons. These included tartarate, quinate, formate, malate, malonate, shikimate, lactate, acetate, maleate, citrate, succinate and fumarate. Soil microbial activity and diversity were determined using dehydrogenase activity and denaturing gradient gel electrophoresis. Links between organic anions in root exudates and rhizosphere soil microbial community structures were investigated by comparing wild type and genetically modified radiata pine trees which were grown in rhizotrons for 10 months. As expected, there was considerable temporal and spatial variability in the amounts and composition of organic anions collected, and there were no consistent or significant differences determined between the two tree lines. Significant differences in rhizosphere microbial communities were detected between wild type and genetically modified pine trees; however, they were inconsistent throughout the experiment. The shifts in microbial communities could have been related to changes in exudate production and composition. Based on results from the main rhizotron experiment, a microcosm study was carried out to investigate the influence of selected pine root exudate sugars (glucose, sucrose and fructose) and organic anions (quinate, lactate and maleate) on soil microbial activity and diversity. Soil microbial activity increased up to 3-fold in all of the sugar and organic anion treatments compared to the control, except for a mixture of sugars and maleate where it decreased. The corresponding impacts on soil microbial diversity were assessed using denaturing gradient gel electrophoresis and 16S rRNA phylochips. Addition of the exudate compounds had a dramatic impact on the composition and diversity of the soil microbial community. A large number of bacterial taxa (88 to 1043) responded positively to the presence of exudate compounds, although some taxa (12 to 24) responded negatively. Organic anions had a greater impact on microbial communities than sugars, which indicated that they may have important roles in rhizosphere ecology of radiata pine. In addition, a diverse range of potentially beneficial bacterial taxa were detected in soil amended with organic anions, indicating specific regulation of rhizosphere microbial communities by root exudates. This project highlighted the considerable challenges and difficulties involved in detailed investigation of in situ rhizosphere processes. Nonetheless, the findings of this study represent a significant contribution to advancing understanding of relationships between root exudates and soil microbial diversity, which will be further enhanced by refinement and application of the specific methodologies and techniques developed.

No description available.

La coiffe racinaire est cruciale à la croissance et survie du méristème subapical de racine. Elle libère des cellules frontières (CFs) qui assurent la protection de l’apex racinaire. Les CFs associées à leur mucilage forment le piège extracellaire de racine (RET). La caractérisation du RET et des exsudats racinaires de trois essences ligneuses sahéliennes à partir de plantules cultivées in vitro a été réalisée. B. aegyptiaca et A. raddiana prospèrent dans les zones semi-arides, à l’opposé de T. indica. La morphologie des CFs et l’organisation du RET ont été déterminées par microscopie. La compostion en glycopolymères et la détection des arabinogalactanes proteines (AGPs) dans le RET et les exsudats racinaires ont été déterminées par des analyses biochimiques. L’effet des exsudats racinaires sur la croissance d’Azospirillum brasilense, une bactérie bénéfique pour la plante a été évalué. B. aegyptiaca produit des CFs de type border cells (BCs) alors que les autres Fabaceae produisent des BCs et des border-like cells. Les BCs sont entourées d’un dense mucilage riche en polymères de paroi. Le RET et les exsudats racinaires issus de B. aegyptiaca et A. raddiana sont plus riches en AGPs que ceux provenant T. indica. Les AGPs pourraient contribuer à la survie des plantules dans un contexte semiaride. Ce travail ouvre de nouvelles perspectives de recherche concernant l'implication du RET dans la survie des plantes à l'aridité. / The root cap is primordial for seedling growth and supports root apical meristem integrity. The root cap releases root border cells (RBCs) that surround the root tip and ensure seedling protection against numerous stresses. RBCs and their associated mucilage form the root extracellular trap (RET). Here, RET and root exudate characterization of three Sahelian woody seedlings are performed. In contrast to B. aegyptiaca and A. raddiana which thrive in semi-arid areas, T. indica is more sensitive to drought. B. aegyptiaca, A. raddiana and T. indica seedlings were sub-cultured in vitro. RBC morphologies and RET organization were determined using microscopic approaches. The polysaccharide composition and arabinogalactan protein (AGP) content were determined by biochemical approaches in the RET and the root exudates. Moreover, the effect of root exudates on the growth of Azospirillum brasilense a plant benefical bacteria has been performed. While B. aegyptiaca produces only border cell (BC) type, the two Fabaceae seedlings release both BCs and border-like cells (BLCs). BCs are enclosed in a dense mucilage enriched in cell wall polymers. Compared to T. indica, RET and root exudates of B. aegyptiaca and A. raddiana include more abundant AGPs. In this context, AGPs could contribute to woody seedling survival. This work opens new research perspectives regarding involvement of RET in plant survival to aridity.

El objetivo de esta Tesis Doctoral es estudiar diferentes herramientas para el manejo de plagas y enfermedades del cultivo de la platanera. Entre las herramientas que vamos a desarrollar, se van a analizar los compuestos orgánicos volátiles (COVs) fúngicos derivados de hongos entomopatógenos (HE) y nematófagos, como fuente de metabolitos con actividad antagónica contra el picudo negro (PN) de la platanera, Cosmopolites sordidus, para su control y manejo en el campo. Así mismo, vamos a estudiar las respuestas de cultivares de plataneras a quitosano, un polisacárido biodegradable, para evaluar su posible uso en el campo como estimulante y protector de las plantas frente a plagas y patógenos, como Fusarium oxysporum f. sp. cubense. El conjunto de capítulos de esta tesis pretende sentar las bases de una estrategia de manejo sostenible de plagas y enfermedades del cultivo de la platanera, basada en el uso de COVs derivados de hongos presentes de forma natural en los cultivos, en combinación con la suplementación de quitosano en el riego, para un efecto de protección y activación de las defensas de las plataneras antes de cualquier infección de plagas y/o enfermedades. El objetivo principal de esta Tesis Doctoral es encontrar nuevas fórmulas para la gestión integrada de plagas como Cosmopolites sordidus y enfermedades de la platanera en condiciones de campo. En esta Tesis Doctoral hemos ideado enfoques sostenibles para la gestión de las plagas y enfermedades de las plataneras. Nuestros objetivos son: a) Cosmopolites sordidus (picudo negro de la platanera, PN), la principal plaga de los cultivos de plátano y, b) el hongo del marchitamiento Fusarium oxysporum f. sp. cubense Raza Tropical 4 (FocTR4), agente causante de una nueva variante extremadamente virulenta de la enfermedad del “Mal de Panamá”, que se está extendiendo rápidamente por todo el mundo. Nuestras herramientas de gestión sostenible son: a) los hongos entomopatógenos (HE, conocidos por su uso como agentes de control biológico, ACBs) aislados de campos comerciales de plátanos, b) sus compuestos orgánicos volátiles (COVs) y, c) el quitosano, un compuesto biodegradable y elicitor de la inmunidad de las plantas con actividad antimicrobiana. Damos evidencia de que los COVs de los hongos agentes de control biológico son repelentes del PN. Pueden utilizarse en los cultivos de platanera mediante estrategias de push and pull para gestionar la plaga de forma sostenible. El quitosano puede utilizarse en el riego para prevenir las defensas de la platanera local y sistémicamente. Por lo tanto, este polímero, con probada actividad antimicrobiana frente a otros patógenos de marchitamiento de Fusarium spp., podría utilizarse contra la actual pandemia en las plataneras causada por FocTR4. La capacidad de inducir reguladores del crecimiento de las plantas sostiene también el papel fertilizante del quitosano. La inducción de compuestos relacionados con la respuesta sistémica adquirida (RSA) hace que el riego con quitosano sea una herramienta para manejar también las plagas de las plataneras sobre el suelo (PN) y las enfermedades (Sigatoka). De esta manera, los COVs y el quitosano podrían ayudar a reducir el uso de agroquímicos tóxicos en los cultivos de platanera en todo el mundo.

Les sols représentent une ressource non renouvelable qui soutient 95% de la production alimentaire mondiale. Cependant, les sols sont de plus en plus impactés par la pollution chimique anthropique, menaçant la santé humaine et l’environnement. Parmi les polluants les plus répandus dans les sols, l'arsenic est aussi l’un des plus dommageables pour la santé humaine, touchant près de 200 millions de personnes dans le monde. Pour limiter la contamination des sols, les approches conventionnelles de remédiation reposent principalement sur l'excavation et l'enfouissement des sols contaminés, mais sont incompatibles avec les grandes surfaces concernées par la contamination chimique, pouvant s’étendre sur des millions d’hectares. Une solution novatrice utilise les plantes et les microorganismes associés pour extraire, dégrader ou stabiliser les contaminants in situ dans une approche dite de phytoremédiation. L'exsudation d'une grande diversité de métabolites des racines dans le sol environnant serait un mécanisme essentiel qui permet aux plantes de tolérer et de détoxifier les contaminants du sol. Cependant, l’environnement chimique de la rhizosphère et les interactions complexes entre les exsudats racinaires et les contaminants restent largement inconnus. L'objectif de cette thèse est de faire progresser la compréhension de l'exsudation racinaire en réponse à la contamination et de son impact sur le devenir de l'arsenic dans la rhizosphère. Le Chapitre 1 rapporte un nouveau système de croissance à petite échelle, hautement reproductible, développé pour capturer et caractériser les exsudats racinaires. En utilisant une analyse métabolomique non ciblée basée sur la chromatographie liquide couplée à la spectrométrie de masse, l’espèce légumineuse Lupinus albus a été examinée pour identifier les différences significatives dans l’exsudation d’un large spectre de composés dans la rhizosphère. Cette approche a révélé les classes prédominantes de composés exsudés et leurs variations en réponse à la contamination, notamment les coumarines, connues pour être impliquées dans les stratégies d'acquisition de nutriments chez les plantes, ainsi que l'exsudation inattendue de phytochélatines, connues pour participer à la complexation et à la détoxification intracellulaire de l'arsenic. Pour confirmer l'exsudation des phytochélatines, une expérience supplémentaire a été menée et a permis de valider l'exsudation des phytochélatines comme mécanisme potentiel de tolérance à l'arsenic dans la rhizosphère. Le Chapitre 2 examine plus en détail les mécanismes d'exsudation des phytochélatines et leurs interactions avec l'arsenic chez Lupinus albus. Grâce à l'inhibition chimique des principales voies de synthèse et d'exsudation des phytochélatines, ce chapitre fournit les premières observations de l’exsudation active de complexes arsenic-phytochélatine chez les plantes, pouvant jouer un rôle critique dans la détoxification de l'arsenic. À partir de ces observations, le chapitre 2 propose une révision du modèle actuel d'efflux d'arsénite des racines et met en évidence l'exsudation de complexes arsenic-phytochélatine comme mécanisme jusque-là inconnu de détoxification chez les plantes. Dans le Chapitre 3, les différentes stratégies employées en réponse à la contamination à l'arsenic chez Lupinus albus et Salix miyabeana sont explorées en profondeur par différents essais en pots et à plus grande échelle, directement sur le terrain. Ces deux espèces, illustrant différents traits fonctionnels importants pour la phytoremédiation, révèlent des adaptations distinctes ainsi que des stratégies d’exsudation conservées en réponse à l'arsenic. Les résultats présentés dans ce chapitre révèlent en particulier le rôle central de l'exsudation de phytochélatines dans la rhizosphère de ces deux espèces, pourtant éloignées phylogénétiquement. Notamment, la découverte de complexes phytochélatine-arsenic dans la rhizosphère des deux espèces souligne l'importance des mécanismes extracellulaires dans la détoxification de l'arsenic chez les plantes. De plus, les mesures sur le terrain soutiennent les implications de l'exsudation des phytochélatines en tant qu’adaptation à l'exposition à l'arsenic en conditions réelles. En résumé, ce chapitre fournit de nouvelles perspectives sur l'interaction complexe entre les plantes et les sols lors de la phytoremédiation de l'arsenic. Dans l'ensemble, cette thèse présente de nouvelles stratégies d'exsudation chez deux espèces phytoremédiatrices majeures et apporte de nouvelles connaissances sur la façon dont l'investissement de ressources dans la rhizosphère peut aider les plantes à tolérer, voire à surmonter, l'effet de la pollution anthropique sur l'environnement. Comprendre ces interactions naturelles est essentiel pour aider à concevoir des stratégies de gestion durables des terres, visant à réduire l'impact à long terme des activités humaines sur les sols. / Soils represent a non-renewable resource supporting 95% of global food production. However, soils face increasing threats from anthropogenic chemical pollution, creating an environmental burden impacting human and environmental health worldwide. Arsenic is one of the most widespread soil contaminants, thought to affect over 200 million people globally and posing substantial threats to public health. To limit contamination of soils, conventional remediation approaches rely on soil excavation and burial, but are incompatible with the extensive problem of soil contamination, often impacting millions of hectares. An innovative solution is to use phytoremediation to harness plants' natural abilities to extract or degrade soil contaminants. The exudation of a wide diversity of metabolites from roots into the surrounding soil is thought to be an essential mechanism used by plants to modify challenging soil environments. However, the extent and variation of root exudation remains largely uncharacterised for many important crops. The objective of this thesis is to advance the understanding of root exudation in response to contamination and how it impacts the fate of arsenic in the rhizosphere. Chapter 1 reports a novel small-scale but highly reproducible growth system developed to capture and characterise root exudates. Using untargeted liquid chromatography-tandem mass spectrometry-based metabolomic analysis, the leguminous crop white lupin (Lupinus albus) was scrutinised to identify significant differences in exuded compounds within the rhizosphere. This approach revealed the predominant classes of exuded compounds in response to contamination, including coumarins, known to be involved in plant nutrient acquisition strategies, as well as unexpected phytochelatin exudation, known to participate in intracellular arsenic complexation and detoxification. A validatory experiment was conducted and confirmed the exudation of phytochelatins as a potential arsenic tolerance mechanism for rhizosphere detoxification. Chapter 2 further investigates the mechanisms of phytochelatin exudation and their interactions with arsenic in Lupinus albus. Through chemical inhibition of key root exudates synthesis and exudation mechanisms, this chapter provides the first evidence that plants actively exude arsenic-phytochelatin complexes, which may function as a critical step for arsenic detoxification and tolerance. From this evidence, Chapter 2 provides a tentative revision of the current model of arsenite efflux from roots and demonstrates that arsenic-phytochelatin exudation may be an active mechanism conferring arsenic tolerance. In Chapter 3, the different strategies employed in response to arsenic contamination in Lupinus albus and Salix miyabeana were extensively scrutinised in larger-scale pot and field trials, to capture the diversity of rhizosphere metabolites within constructed and real-world soils. These species, illustrating different important functional traits for phytoremediation, revealed distinct as well as more conserved root exudate adaptations to arsenic. Most importantly, the findings presented in this chapter reveal a conserved and pivotal role for extracellular phytochelatin exudation in the rhizosphere of these distantly related phytoremediating species. The discovery of phytochelatin-arsenic complexes in the rhizosphere of both species underscores the importance of extracellular mechanisms in plant arsenic detoxification. Furthermore, field assessments supported the real-world implications of phytochelatin exudation as an adaptive response to arsenic exposure. In summary, this chapter provides novel insights into the complex interplay between plants and soils in arsenic phytoremediation. Overall, this thesis presents novel exudation strategies in two major phytoremediation species and brings new knowledge on how investment of resources in the rhizosphere can help plants tolerate, or even overcome, the effect of anthropogenic pollution upon the natural environment. Understanding these mechanisms is vital to devise sustainable land management strategies to reduce the long-term impact of human activity on soils around the world.