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Bioprospecting di simbionti vegetali con proprietà PBS per lo sviluppo di nuovi prodotti biostimolanti: bridging tra i risultati della ricerca e gli aspetti normativi. / BIOPROSPECTING OF PLANT SYMBIONTS WITH PBS PROPERTIES FOR THE DEVELOPMENT OF NOVEL PLANT BIOSTIMULANT PRODUCTS: BRIDGING RESEARCH OUTCOMES WITH REGULATORY ASPECTSGUERRIERI, MARIA CHIARA 28 April 2021 (has links)
L'agricoltura moderna sta affrontando sfide come la perdita di fertilità del suolo, la variabilità climatica e gli attacchi di agenti patogeni in continuo aumento. Le pratiche agricole si stanno evolvendo verso sistemi sostenibili e rispettosi dell'ambiente. L'uso di biostimolanti (PBS, plant biostimulant) è una soluzione innovativa per affrontare le sfide di un’agricoltura sostenibile che garantisce un assorbimento ottimale dei nutrienti, una resa delle colture e tolleranza agli stress abiotici. In particolare, tra i diversi tipi di biostimolanti presenti sul mercato, i rizobatteri, classificati come Plant Growth Promoting Rhizobacteria (PGPR), offrono un nuovo approccio per promuovere la crescita delle piante, la mitigazione degli stress e l’aumento della resa colturale. Pertanto i PGPR sono considerati come una sorta di "probiotici" vegetali, poiché contribuiscono in modo efficiente alla nutrizione e all'immunità delle piante. L'obiettivo principale di questa tesi è isolare e identificare batteri presenti nella rizosfera di pomodoro (Solanum lycopersicum L.) che mostrano proprietà PBS, nonché valutare i meccanismi coinvolti nell'azione di promozione della crescita delle piante (Capitolo 2) e la genetica alla base di questi meccanismi (Capitolo 3 e 4). Infatti, una profonda comprensione dei meccanismi d’azione dei PGPR potrebbe colmare la mancanza di coerenza del dato di efficacia tra gli studi di laboratorio e gli studi in campo e stimolare la ricerca per la produzione e la commercializzazione di nuovi prodotti biostimolanti microbici. / Modern agriculture faces challenges such as loss of soil fertility, fluctuating climatic factors and increasing pathogen and pest attacks. Agricultural practices have been evolving towards organic, sustainable and environmentally friendly systems. The use of natural plant biostimulants (PBS) is an innovative solution to address the challenges in sustainable agriculture, to ensure optimal nutrient uptake, crop yield, quality and tolerance to abiotic stress. In particular, among different types of biostimulants present on the market, plant growth promoting rhizobacteria (PGPR) offer a novel approach for promoting plant growth, mitigate stress and increase crop yield. Hence, PGPR inoculants are now considered as a kind of plant ‘probiotics’, since they efficiently contribute to plant nutrition and immunity. The main goal of this thesis was to isolate and identify bacteria symbionts of tomato (Solanum lycopersicum L.) rhizosphere, which showed PBS properties and evaluate mechanism involved in the action of PGPR (Chapter 2), underlying genetics and physiological pathways (Chapter 3 and 4). Indeed, a deeply understanding of the mechanisms of plant growth promotion, could fulfill the lack of consistency between lab, greenhouse and field studies, and support commercialization of novel plant biostimulant products.
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Pseudomonas spp. Isolated from Soybean Nodules Promote Soybean Growth and Nitrogen FixationGriggs, Roland Stephen 08 June 2020 (has links)
Nitrogen-fixing bacteria in soybean nodules convert atmospheric nitrogen to plant-available forms in exchange for carbon from the plant, but other non-nitrogen-fixing bacteria also reside in nodules, and their role in the nodule is not well understood. This study was conducted to determine the effect of three non-nitrogen-fixing Pseudomonas spp. strains isolated from nodules on soybean, and we hypothesized these strains benefit soybean. A greenhouse study in which two cultivars of soybean (Asgrow AG46X6 and Pioneer P48A60X) were treated with three fluorescent Pseudomonas spp. strains (referred to in this study as Bullseye, Pancake, and Starfish) and an uninoculated control. Soybeans were harvested at two time points: the R2/R3 growth stage and the R6 growth stage. Following each harvest, measures of growth, yield, and nitrogen fixation were taken, and data were analyzed using two non-parametric, multivariate analyses: multiple response permutation procedure (MRPP) and permutational multivariate analysis of variance (PERMANOVA). Both analyses showed soybeans of both cultivars treated with Pancake differed from controls following the first harvest but not the second. When analyzed individually, most metrics for growth, yield, and nitrogen fixation following the first harvest were not significantly different between Pancake and control treatments, but Pancake treatment means were still generally higher than controls. If metrics are considered collectively in conjunction with the results of the multivariate analyses, the results show Pancake generally increased soybean growth and nitrogen fixation. These findings support the hypothesis that non-nitrogen-fixing bacteria from nodules benefit plants, and such bacteria have the potential to serve as biofertilizers. / Master of Science in Life Sciences / Soybeans are one of the most commonly grown crops in the world, and nitrogen-fixing bacteria colonize the roots of soybeans and initiate the formation of spherical nodules attached to the roots. Inside the nodules, these bacteria convert atmospheric nitrogen to plant-available forms in exchange for sugar from the plant, and such bacteria reduce the need to add nitrogen fertilizer to agricultural fields. Other non-nitrogen-fixing bacteria also reside in nodules, but their role in the nodule is not well understood. If these bacteria benefit soybeans, they have the potential to serve as biofertilizers (microbial inoculants that promote plant growth). This study was conducted to determine whether non-nitrogen-fixing bacteria isolated from nodules benefit soybean. A greenhouse study in which two cultivars of soybean (Asgrow AG46X6 and Pioneer P48A60X) were grown in soil and were either left uninoculated or were inoculated with one of three strains of bacteria from the genus, Pseudomonas (referred to in this study as Bullseye, Pancake, and Starfish). Following harvest, measures of growth, yield, and nitrogen fixation were taken, and data showed the bacteria generally benefited the soybean plants. Although, these results showed the bacteria benefitted the plants, field trials and further testing in the greenhouse should be conducted before using these bacteria as commercial biofertilizers. Additionally, the effects of other non-nitrogen-fixing nodule bacteria on soybeans should also be tested to identify other beneficial strains, and the cost of production should be compared to the potential gains of using such bacteria before they are developed into biofertilizers.
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Pseudomonas spp. Isolated from the Soybean Nodule Interior Promote Soybean Growth upon Field AmendmentDoyle, Connor Patrick 31 August 2022 (has links)
Diazotrophic microbes reside in soybean nodules; however, other non-nitrogen fixing bacteria are a part of the interior nodule microbiome. Results from a previous greenhouse study show that a novel species of Pseudomonas associates with soybean nodules as a plant-growth promoting rhizobacteria (PGPR). This study observes the soybean growth promoting potential of Pseudomonas spp. in a field setting. Additionally, this study observed differences in soybean growth promotion based on amending the plant with isolated strains or a mixed culture of the species' strains. Two cultivars of soybean (Asgrow AG46X6 and Pioneer P48A60X) were either amended with isolated strains of the novel Pseudomonas spp. (referred to as PAMW1 and BUMW2 in this study), a mix of the two strains, or an uninoculated control. The study recorded measurements to observe growth, yield, and nitrogen fixation differences. The study uses two-way factorial ANOVAs and non-parametric, multivariate analyses to determine differences in growth promotion among samples. Soybean amended with PAMW1 has greater shoot mass, biomass, and height than other treatments. Through nonmetric multidimensional scaling (NMS), samples amended with a mixed culture or PAMW1 may be different regarding growth promotion relative to the non-amended samples. Univariate results support the hypothesis that the novel Pseudomonas spp. benefit soybean in a field setting. However, it is inconclusive whether a mixed culture amendment of multiple strains alters the overall growth promotion of soybean compared to samples amended with isolated strains. / Master of Science / Soil hosts a relatively abundant and diverse community of microorganisms. Moreover, the area of soil that interacts closely with plant roots and their associated exudates, called the rhizosphere, has a significantly greater microbial abundance than surrounding bulk soil. Interactions between microbes and the plant often promote plant growth because of secondary metabolites produced by these beneficial microbes. One particular bacterial species, belonging to the Pseudomonas genus, was discovered and extracted from the soybean nodule interior. Nitrogen-fixing bacteria predominantly reside in the soybean nodule, yet this microorganism cannot fix nitrogen. Although trace amounts of non-nitrogen-fixing bacteria reside in the soybean nodule, this novel species has a relatively high abundance. This study determines the benefits of this species in the soybean nodule. Following positive results in a greenhouse study, this field experiment observes variance in soybean growth and productivity based on their received bacterial amendment. For this study, two soybean cultivars were either amended with an isolated strain of this species, a mix of the two strains, or left uninoculated to serve as a control. Numerous recorded measurements serve as indices of soybean growth and productivity. The results suggest that this novel Pseudomonas species benefits the plant by significantly improving biomass. With further research, this species can potentially serve as an environmentally sensitive and sustainable alternative to fertilizers through its ability to promote soybean growth.
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Étude des communautés microbiennes rhizosphériques de ligneux indigènes de sols anthropogéniques, issus d’effluents industriels / Study of rhizosphere microbial communities from native woody species collected on anthropogenic soils made of industrial effluentsZappelini, Cyril 03 July 2018 (has links)
Mon sujet de thèse intègre l’un des projets globaux de l’UMR UFC/CNRS 6249 Chrono-Environnement intitule « stratégies de phytoremédiation basées sur l’utilisation d’arbres et de microorganismes associés », qui s’appuie, entre autre, sur 2 projets de recherche :• le projet PROLIPHYT (programme Eco-Industrie, 2013-2018, ADEME) intitulé « PROduction de LIgneux PHYtoremédiants»,• le projet PHYTOCHEM (ANR CD2i, 2013-2018) intitulé « Développement de procédés chimiques éco-innovants pour valoriser les biomasses issues des phytotechnologies ».Les objectifs généraux sont d’améliorer le potentiel de phytoremédiation d’un panel d’espèces ligneuses et de développer le potentiel microbien pour une phytoremédiation aidée sur sol contaminé. En plus de limiter l’impact des polluants, cette stratégie vise à promouvoir la production de biomasse sur sols délaissés et non exploitables par l’agriculture, tout en assurant la biodiversité nécessaire à la restauration d’un écosystème anthropogénique.Mon travail de thèse est financé au travers un contrat doctoral ministériel handicap (dyslexie). Il s’appuie sur la réhabilitation de deux zones de stockage de sédiments industriels, utilisés jusque dans les années 2000. Ces deux sites expérimentaux (site INOVYN de St Symphorien-sur-Saône en Côte d’Or, site CRISTAL de l’Ochsenfeld en Alsace) présentent des caractéristiques physico-chimiques très particulières qui en font des lieux d’étude privilégiés. Le premier est une ancienne lagune de décantation dont les sédiments enrichis en Hg, Ba et As proviennent du traitement des eaux usées issues du procédé d’électrolyse à Hg de l’entreprise SOLVAY. Le second est une lagune constituée d’un remblai dans lequel ont été stockés depuis les années 1930, les résidus d’extraction du dioxyde de titane de l’Usine CRISTAL de Thann. A l’inverse du premier site expérimental, on observe une flore peu abondante qui se traduit par un développement hétérogène d’une espèce ligneuse principale, le bouleau.La recolonisation naturelle et spontanée de végétaux, plus particulièrement d’espèces ligneuses sur les deux sites est sans doute le résultat d’étroites collaborations avec des microorganismes telluriques situés aux abords de leur système racinaire. Nous avons ainsi choisi de travailler sur 3 espèces pionnières qui se sont naturellement réimplantées sur les deux sites d’études : le saule et le peuplier pour la friche industrielle de Tavaux et le bouleau pour l’unité de traitement des effluents du site de l’Ochsenfeld. / AbstractMy thesis subject includes one of the global projects of the UMR UFC/CNRS 6249 Chrono-Environnement entitled "phytoremediation strategies based on the use of trees and associated microorganisms", which is based, among other things, on 2 research projects:• the PROLIPHYT project (Eco-Industry programme, 2013-2018, ADEME) entitled "Production of woody phytoremediants",• the PHYTOCHEM project (ANR CD2i, 2013-2018) entitled "Development of eco-innovative chemical processes to exploit biomasses from phytotechnologies".The general objectives are to improve the phytoremediation potential of a panel of woody species and to develop the microbial potential for assisted phytoremediation on contaminated soil. In addition to limiting the impact of pollutants, this strategy aims to promote the production of biomass on land abandoned and not exploitable by agriculture, while ensuring the biodiversity needed to restore an anthropogenic ecosystem.My thesis work is financed through a ministerial doctoral contract for disability (dyslexia). It is based on the rehabilitation of two industrial sediment storage areas, used until the 2000s. These two experimental sites (INOVYN site of Saint-Symphorien-sur-Saône in Côte-d'Or, CRISTAL site of Ochsenfeld in Alsace) present very particular physico-chemical characteristics which make them privileged places of study. The first is a former settling lagoon whose sediments enriched in Hg, Ba and As come from the treatment of wastewater from SOLVAY's Hg electrolysis process. The second is a lagoon consisting of a backfill in which the titanium dioxide extraction residues from the CRISTAL Thann Plant have been stored since the 1930s. In contrast to the first experimental site, there is a low abundance of flora which results in heterogeneous development of a main woody species, the birch.The natural and spontaneous recolonisation of plants, more particularly woody species on both sites, is undoubtedly the result of close collaboration with telluric microorganisms located near their root systems. We have thus chosen to work on 3 pioneer species that have naturally relocated to the two study sites: willow and poplar for the industrial wasteland of Tavaux and birch for the effluent treatment unit at the Ochsenfeld site.
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TOXICITY OF ENGINEERED NANOMATERIALS TO PLANT GROWTH PROMOTING RHIZOBACTERIALewis, Ricky W. 01 January 2016 (has links)
Engineered nanomaterials (ENMs) have become ubiquitous in consumer products and industrial applications, and consequently the environment. Much of the environmentally released ENMs are expected to enter terrestrial ecosystems via land application of nano-enriched biosolids to agricultural fields. Among the organisms most likely to encounter nano-enriched biosolids are the key soil bacteria known as plant growth promoting rhizobacteria (PGPR). I reviewed what is known concerning the toxicological effects of ENMs to PGPR and observed the need for high-throughput methods to evaluate lethal and sublethal toxic responses of aerobic microbes. I addressed this issue by developing high-throughput microplate assays which allowed me to normalize oxygen consumption responses to viable cell estimates. Oxygen consumption is a crucial step in cellular respiration which may be examined relatively easily along with viability and may provide insight into the metabolic/physiological response of bacteria to toxic substances.
Because many of the most toxic nanomaterials (i.e. metal containing materials) exhibit some level of ionic dissolution, I first developed my methods by examining metal ion responses in the PGPR, Bacillus amyloliquefaciens GB03. I found this bacterium exhibits differential oxygen consumption responses to Ag+, Zn2+, and Ni2+. Exposure to Ag+ elicited pronounced increases in O2 consumption, particularly when few viable cells were observed. Also, while Ni2+ and Zn2+ are generally thought to induce similar toxic responses, I found O2 consumption per viable cell was much more variable during Ni2+ exposure and that Zn2+ induced increased O2 utilization to a lesser extent than Ag+. Additionally, I showed my method is useful for probing toxicity of traditional antibiotics by observing large increases in O2 utilization in response to streptomycin, which was used as a positive control due to its known effects on bacterial respiration.
After showing the utility of my method for examining metal ion responses in a single species of PGPR, I investigated the toxicity of silver ENMs (AgENMs) and ions to three PGPR, B. amyloliquefaciens GB03, Sinorhizobium meliloti 2011, and Pseudomonas putida UW4. The ENM exposures consisted of untransformed, polyvinylpyrrolidone coated silver ENMs (PVP-AgENMs) and 100% sulfidized silver ENMs (sAgENMs), which are representative of environmentally transformed AgENMs. I observed species specific O2 consumption responses to silver ions and PVP-AgENMs. Specifically, P. putida exhibited increased O2 consumption across the observed range of viable cells, while B. amyloliquefaciens exhibited responses similar to those found in my first study. Additionally, S. meliloti exhibited more complex responses to Ag+ and PVP-AgENMs, with decreased O2 consumption when cell viability was ~50-75% of no metal controls and increased O2 consumption when cell viability was <50%. I also found the abiotically dissolved fraction of the PVP-AgENMs was likely responsible for most of the toxic response, while abiotic dissolution did not explain the toxicity of sAgENMs.
My work has yielded a straightforward, cost-effective, and high-throughput method of evaluating viability and oxygen consumption in aerobic bacteria. I have used this method to test a broad range of toxic substances, including, metal ions, antibiotics, and untransformed and transformed ENMs. I observed species specific toxic responses to Ag+, PVP-AgENMs, and sAgENMs in PGPR. These results not only show the clear utility of the methodology, but also that it will be crucial to continue examining the responses of specific bacterial strains even as nanotoxicology, as a field, must move toward more complex and environmentally relevant systems.
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Comprehensive proteomic study of Bacillus amyloliquefaciens strain FZB42 and its response to plant root exudatesKierul, Kinga 19 August 2013 (has links)
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.
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Interaction plante-microorganismes : Implication de la rhizobactérie Phyllobacterium brassicacearum dans les réponses d’Arabidopsis thaliana au stress hydrique / Plant-microbes interactions : Implication of Phyllobacterium brassicacearum in Arabidopsis responses to water deficitBresson, Justine 16 December 2013 (has links)
Les bactéries promotrices de la croissance des plantes (PGPR) peuvent améliorer la performance et la tolérance des plantes lors de stress environnementaux. Arabidopsis thaliana est un modèle de choix pour étudier les mécanismes impliqués dans les interactions plante-bactéries. Nous avons analysé de multiples traits associés à la dynamique de croissance, au développement et la physiologie des végétaux afin d'évaluer les effets de l'inoculation par Phyllobacterium brassicacearum STM196, une PGPR isolée de la rhizosphère du colza, sur les réponses d'A. thaliana à des stress hydriques de différentes intensités. Grâce à des outils performants de phénotypage, nous avons développé une nouvelle approche d'analyse à haut-débit pour examiner l'implication de STM196 dans les stratégies de résistance des plantes au stress hydrique. Nos résultats montrent pour la première fois que les PGPR peuvent interférer dans les stratégies d'échappement des plantes grâce à des modifications de la croissance et du temps de floraison. De plus, STM196 induit une meilleure résistance au déficit hydrique modéré et une meilleure tolérance à la déshydratation sous une contrainte hydrique sévère. L'inoculation par STM196 peut ainsi représenter une valeur ajoutée aux stratégies de résistance intrinsèques aux plantes, ce qui est illustrée par sa remarquable capacité à promouvoir la survie et la production de biomasse végétale dans des environnements contrastés. Nos résultats soulignent l'importance des interactions plantes-bactéries dans les réponses des plantes à la sécheresse et offrent de nouvelles voies de recherches pour l'amélioration de la résistance à la sécheresse dans les cultures. / Plant growth promoting rhizobacteria (PGPR) can enhance plant performance and plant tolerance to environmental stresses. Arabidopsis thaliana is a useful organism to study the mechanisms involved in plant-PGPR interactions. We analyzed multiple plant traits related to growth dynamics, development and physiology in order to assess the effects of Phyllobacterium brassicacearum STM196 strain, isolated from the rhizosphere of oilseed rape, on Arabidopsis responses to well-defined soil water availability. Using powerful tools for phenotyping, we developed a new high-throughput analysis to examine the implication of STM196 on plant strategies to cope with water stress. Our results show for the first time that PGPR can interfere in escape strategies of plants through modifications in plant growth and flowering time. Moreover, STM196 induced a better resistance to moderate water deficit and a better tolerance to dehydration under a severe stress. Inoculation by STM196 can represent an added value to plant resistance strategies, as illustrated by its remarkable ability to promote plant survival and biomass production under contrasted environments. Our results highlight the importance of plant-bacteria interactions in plant responses to drought and provide a new avenue of investigations to improve drought resistance in crops.
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Identification and characterization of type III effector proteins in plant-associated bacteriaThomas, William J. 04 May 2012 (has links)
Symbioses between microbes and multicellular eukaryotes are found in all biomes, and encompass a spectrum of symbiotic lifestyles that includes parasitism and disease, commensalism, and mutually beneficial interdependent host-microbe relationships. Regardless of outcome, these symbiotic lifestyles are governed by a complex molecular "courtship" between microbe and potential host. This courtship is the primary determinant of the host range of a given microsymbiont. Host immunity poses a formidable barrier to the establishment of host-microbe relationships, and the majority of microbial suitors will be thwarted by it. Only by successfully "wooing" the host cell's immune defenses with the appropriate molecular signals can a microsymbiont successfully colonize its host.
A strategy common to microsymbionts across the spectrum of symbiotic lifestyles and host organisms is the delivery of microbial-encoded effector proteins into the cytoplasm of host cells to manipulate the host cell's molecular machinery for the purposes of subverting host immunity. Bacteria, in particular, have adapted a number of secretion systems for this purpose. The most well-characterized of these is the type III secretion system (T3SS), a molecular apparatus that specializes in injecting type III effector (T3Es) proteins directly into host cells. The work in this thesis focuses on T3Es of plant-associated bacteria, with particular emphasis on mutualistic bacteria. We present evidence that collections of T3Es from Sinorhizobium fredii and Bradyrhizobium japonicum are, in stark contrast to those of phytopathogenic bacteria, in a co-evolutionary equilibrium with their hosts. This equilibrium is characterized by highly conserved T3E collections consisting of many "core" T3Es with little variation in nucleotide sequence. The T3Es of Mesorhizobium loti MAFF303099 suggest a completely different picture of the evolution of T3Es. MAFF303099 recently acquired its T3SS locus, and the work in this thesis provides an evolutionary snapshot of a mutualist that is innovating a T3E collection primarily through horizontal gene transfer. Collectively, this work represents the first comprehensive catalog of T3Es of rhizobia and, in the case of Sinorhizobium and Bradyrhizobium, the first evidence of purifying selection for T3Es. / Graduation date: 2012
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Improving abiotic and biotic stress tolerance in floriculture cropsSouth, Kaylee 13 November 2020 (has links)
No description available.
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Untersuchungen über die Wirkung von Stoffwechselprodukten, insbesondere Auxinen, des wachstumsfördernden Rhizobakteriums (PGPR) Bacillus subtilis auf die pflanzliche SalztoleranzStavropoulou, Archontia 04 August 2005 (has links)
Zur Aufklärung des Wirkungsmechanismus der toleranzerhöhenden Wirkung gegenüber Salinität des Pflanzenwurzeln besiedelnden PGPR Bacillus subtilis wurden bakterielle Stoffwechselprodukte der Stämme FZB24 und FZB41 bei der Testpflanze Tomate unter dem Einfluss von hohem Salzstress getestet. Das Kulturfiltrat mit der Gesamtheit der von B. subtilis produzierten Stoffwechselprodukte zeigte im axenischen Test zur Ermittlung des Wachstums nach 7-tägiger Behandlung der Sämlinge und nachfolgender Kultivierung unter Salzstress eine gewisse toleranzerhöhende Wirkung bei 0,1 %-Konzentration. Zur Produktaufschlüsselung wurde das Kulturfiltrat über Adsorberharz und HPLC fraktioniert. Diese Fraktionen, sowie die aus dem Kulturfiltrat nach 19 h Fermentation wurden ebenfalls bei Sämlingen axenisch getestet. Fraktionen mit verschiedenen Proteinen und Peptiden, die von B. subtilis produziert werden, zeigten teilweise eine konzentrationsabhängige Wirkung hinsichtlich der Wachstumsstimulierung und zugleich Toleranzerhöhung gegenüber Salzstress, weshalb nachfolgend ein Peptidextrakt aus B. subtilis einer Testung im axenischen System unterzogen wurde. Der Peptidextrakt zeigte gleichfalls eine erkennbare konzentrationsabhängige Wirkung. Mit gleichem Testsystem wurden Auxin-Präkursoren und Auxin selbst, die als Stoffwechselprodukte von B. subtilis nachgewiesen sind, sowohl als Wurzelbehandlung, wie auch als Blattbehandlung bei Sämlingen geprüft. Zusätzlich wurde die Wirkung der Auxine auf den Wassergehalt der Sämlinge unter Salzstress, sowie die Adventivwurzelbildung von Hypokotylsegmenten aus etiolierten Sämlingen in An- und Abwesenheit von Salinität getestet. Darüber hinaus wurde die Aufnahme und der Transport von Auxinen, ebenfalls bei Sprosssegmenten aus etiolierten Sämlingen in An- und Abwesenheit von Salinität geprüft. Schließlich wurde die Wirkung der Auxine auf das Wachstum und den Wassergehalt in einer Hydrokultur im Gewächshaus unter Salzstress ermittelt. Die Ergebnisse zeigen, dass namentlich Auxin-Präkursoren und z. T. Auxin als Stoffwechselprodukte von B. subtilis eine Erhöhung der Salzstresstoleranz bei der Testpflanze herbeiführen können, wenngleich die Wirkung auf die Salztoleranz sehr differenziert und unterschiedlich stark ausgeprägt war. Der vorhandene Effekt vor allem der Auxin-Präkursoren wird als offenbar bedeutendster Mechanismus für die wachstumsstimulierende und zugleich toleranzerhöhende Wirkung gegenüber Salinität des Rhizobakteriums bei Wurzelbesiedlung und Interaktion mit dem pflanzlichen Stoffwechsel diskutiert. / To find out the mode of tolerance increasing action against salinity of the plant root colonizing PGPR Bacillus subtilis, bacterial metabolites of the strains FZB24 and FZB41 were studied in the test plant tomato under the influence of high salinity. Because the culture filtrate with the whole range of produced metabolites by B. subtilis showed to a certain extent a tolerance increasing action at dilution of 0,1 % in axenic plant growth tests after 7 days treatment of seedlings and subsequent cultivation under salt stress, it has been fractionated with adsorber resin and HPLC. These fractions, as well as fractions from the culture filtrate after 19 h fermentation were tested also by seedlings in axenic culture. Fractions with different proteins and peptides, which were produced by B. subtilis, showed partly activities also depending of concentration with regard to the growth stimulation and at the same time tolerance increase against salt stress. Following also a peptide extract from B. subtilis was examined in the axenic plant test system, showing similarly a visible action depending of concentration. In the same test system there were tested further auxin precursors and auxin itself, which are known metabolites of B. subtilis, on seedlings both by root treatment and leaf treatment. Additionally was studied the action of auxins on the water content of the seedlings under salt stress, as well as on the adventitious root formation of hypokotyl segments from etiolated seedlings, in presence and absence of salinity. Finally it was studied the uptake and transport of auxins in segments of stems from etiolated seedlings in presence and absence of salinity. Lastly it was tested the action of auxins on plant growth and water content in a hydroponic cultivation under greenhouse conditions and salt stress. The results show that particularly auxin precursors and partly auxin as metabolites of B. subtilis can induce an increase in the salt stress tolerance of the test plant, although the action on the salt tolerance was differentiated and variable in its extent. The existing effect firstly of the auxin precursors is discussed as obviously main mechanism for the plant growth stimulating and at the same time tolerance increasing action of the rhizobacterium against salinity by root colonization and interaction with the plant metabolism.
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