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UNVEILING NOVEL ASPECTS OF D-AMINO ACID METABOLISM IN THE MODEL BACTERIUM PSEUDOMONAS PUTIDA KT2440Radkov, Atanas D. 01 January 2015 (has links)
D-amino acids (D-AAs) are the α-carbon enantiomers of L-amino acids (L- AAs), the building blocks of proteins in known organisms. It was largely believed that D-AAs are unnatural and must be toxic to most organisms, as they would compete with the L-counterparts for protein synthesis. Recently, new methods have been developed that allow scientists to chromatographically separate the two AA stereoisomers. Since that time, it has been discovered that D-AAs are vital molecules and they have been detected in many organisms. The work of this dissertation focuses on their place in bacterial metabolism. This specific area was selected due to the abundance of D-AAs in bacteria-rich environments and the knowledge of their part in several processes, such as peptidoglycan synthesis, biofilm disassembly, and sporulation. We focused on the bacterium Pseudomonas putida KT2440 which inhabits the densely populated plant rhizosphere. Due to its versatility and cosmopolitan character, this bacterium has provided an excellent system to study D-AA metabolism.
In the first chapter, we have developed a new approach to identify specific genes encoding enzymes acting on D-AAs, collectively known as amino acid racemases. Using this novel method, we identified three amino acid racemases encoded by the genome of P. putida KT2440. All of the enzymes were subsequently cloned and purified to homogeneity, followed by a complete biochemical characterization. The aim of the second chapter was to understand the specific role of the peculiar broad-spectrum amino acid racemase Alr identified in chapter one. After constructing a markerless deletion of the cognate gene, we conducted a variety of phenotypic assays that led to a model for a novel catabolic pathway that involves D-ornithine as an intermediate. The work in chapter three identifies for the first time numerous rhizosphere-dwelling bacteria capable of catabolizing D-AAs. Overall, the work in this dissertation contributes a novel understanding of D-AA catabolism in bacteria and aims to stimulate future efforts in this research area.
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Adaptation de la symbiose rhizobienne chez le haricot à la déficience en phosphore : détermination de la réponse de la plante en terme d'échanges gazeux et de flux minéraux échangés avec la rhizosphère / Nodulated bean adaptation to P deficiency : determination of nodule respiration and proton effluxAlkama, Nora 09 December 2010 (has links)
La déficience en phosphore est l’un des facteurs limitant la production de légumineuses à graines, dont le haricot (Phaseolus vulgaris). La diversité génétique de cette espèce en Amérique latine a permis d'identifier avec le CIAT, des lignées recombinantes du croisement de DOR364 et BAT477 à fort pouvoir fixateur d'azote (FSN) et à forte efficacité d'utilisation de phosphore (EUP). Les résultats obtenus au cours de ce travail nous ont permis de répondre à notre objectif principal de thèse qui consistait à vérifier la pertinence des critères d'adaptation de la fixation symbiotique de l'azote chez le haricot à la disponibilité de P que nous avons déterminé. Nous avons démontré, d'une part, en milieu hydroaéroponique, que les racines nodulées du haricot, excrètent dans leur rhizosphère une quantité de H+ qui est corrélée à la perméabilité nodulaire. Ce qui laisse penser qu'une part d'H+ excrétée par les racines nodulées est liée à la fixation symbiotique de N2. D'autre part, nous avons démontré que sous déficience en P les lignées tolérantes acidifient plus leur rhizosphère que les sensibles. En multisites les facteurs les plus déterminants de la hiérarchisation des sites sont N-total et P-total. Deux groupes de lignées se distinguent, les tolérantes versus les sensibles à la faible fertilité des sols en P. La lignée locale se distingue des groupes précédents. Elle est capable de croître dans différents sols, notamment les contraignants avec une grande capacité à noduler, par conséquent, à compenser la déficience en N. Nous avons également pu démontrer que les biomasses nodulaire et aérienne sont corrélées au P Olsen du sol / Phosphorus deficiency is one of the limiting factor for grain legume production like bean (Phaseolus vulgaris). The results obtained during this work enabled us to confirm our main aim of this work which is checking the relevance of the criteria of adaptation of the symbiotic nitrogen fixation of bean to P availability. We showed, in controlled conditions that the nodulated roots of bean, release in their rhizosphere a quantity of H+ which is correlated with the nodul permeability. What lets think that a share of H+ released is related to the symbiotic N2 fixing. In addition, we showed that under P deficiency the tolerant lines acidify more their rhizosphere that the sensitive ones. Into multisite trial the most determining factors of the hierarchisation of the sites are total-N and total-P. Two groups of lines are distinguished: tolerant versus sensitive to P deficiency. The local farmer line is distinguished from the studied lines. It is able to grow in various soil fertility, in particular the constraining soils. ones with a great capacity with noduler, consequently, to compensate for deficiency in NR. We also could show that shoot and nodule biomass are correlated with the Olsen-P Olsen of the soil
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Caractérisation de la communauté fongique impliquée dans la minéralisation du soufre organique dans les rhizosphères de colza et d'orge / Characterization of fungal community implicated in the mineralization of organic sulfur in rhizosphere of rape and barleyHamdan, Lama 09 November 2010 (has links)
En Europe de l’Ouest, S est devenu un élément limitant pour la croissance des plantes. Ainsi, des carences en S apparaissent de plus en plus fréquemment sur des cultures tel que le colza. Dans le sol, 95% de S est sous formes organiques, non disponibles pour les plantes. L’intervention de la microflore est indispensable pour assurer la minéralisation du S organique en sulfates, assimilables par la plante. Nos objectifs ont été de caractériser la communauté fongique impliquée dans la minéralisation des esters de S, forme majoritaire de S organique, via une activité arylsulfatase (ARS), dans les rhizosphères de colza et d’orge. La communauté fongique est composée de plusieurs genres affiliés principalement aux Ascomycètes. Chez les souches fongiques isolées de la rhizosphère de colza et d’orge, une activité ARS a été détectée dans différents compartiments cellulaires. La régulation de ces activités ARS semble dépendante du taxon considéré. Nous avons par ailleurs montré que l’environnement rhizosphérique n’influence pas toujours la taille de la communauté fongique ARS. Dans les sols, si les activités ARS totale et intracellulaire semblent négativement corrélées avec les quantités de sulfates, l’activité ARS extracellulaire semble indépendante de la disponibilité en sulfates. En conclusion, l’ensemble des expérimentations suggère que la communauté fongique fonctionnelle joue un rôle dans la dynamique du S dans les sols agricoles. Des approches d’écologie fonctionnelle permettraient de mieux cerner leur implication dans la disponibilité en S minéral pour la plante / In Western Europe, sulfur (S) deficiency occurs in certain crops, including crucifers and cereals. Therefore, S becomes limiting for crop production and plants exhaust S mainly from soil organic S. In soil, 95% of S is in organic form that is not readily available for plants. This organic form containing principally ester S requires microbial mineralization to sulfate by arylsulfatase (ARS) enzyme. Our objectives were to characterize the fungal community having the ARS activity in the rhizosphere of rape versus that of barley. Functional fungal community comprised several genera principally belonging to Ascomycota. In different fungal strains, ARS activity was detected in different cellular compartments. The regulation of ARS was mostly dependent on microbial taxa. The density of the functional fungal community was not influenced by rhizospheric compartment.In soils, total and intracellular ARS activities were negatively correlated with soil sulfates whereas soil extracellular ARS activity was independent of sulfates. The overall results suggest that the functional fungal community could play a role in the dynamics of S in agricultural soils. Further approaches should be developed to allow a better understanding of their potential involvement in S nutrition of crops
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Impact de signaux rhizosphériques sur l’expression de gènes phytobénéfiques chez les bactéries symbiotiques associatives / Impact of signalling rhizospheric molecules on the expression of plant beneficial genes from associative symbiotic bacteriaCombes-Meynet, Émeline 08 April 2010 (has links)
La rhizosphère est le siège de multiples interactions biotiques, contrôlées au moins en partie par l’échange de molécules signal entre les racines des plantes et les microorganismes qui leur sont associés. Dans le contexte des symbioses associatives entre les PGPR (Plant-Growth Promoting Rhizobacteria) et les plantes, le rôle et l’importance de ces molécules restent encore largement méconnus. Ainsi, l’influence de différents signaux rhizosphériques sur l’expression génique et les propriétés phytobénéfiques des PGPR a été étudiée par, (i) la caractérisation chimique d’une nouvelle molécule phytobénéfique d’Azospirillum et de son impact sur l’architecture du système racinaire, (ii) une approche ciblée portant sur l’analyse de l’expression in situ du gène phytobénéfique acdS de PGPR, et (iii) une approche globale permettant d’identifier la réponse du transcriptome d’Azospirillum à un métabolite secondaire bactérien. Ces travaux ont permis d’identifier des signaux rhizosphériques impliqués dans le contrôle de la colonisation racinaire et/ou des propriétés de phytostimulation des PGPR sur leur plante hôte / In rhizosphere, numerous biotic interactions are controlled by molecular signals exchanged between plant roots and microorganisms. In the case of the associative symbiosis between PGPR (Plant-Growth Promoting Rhizobacteria) and plants, the role and importance of these molecules are largely unknown. The impact of several rhizospheric signals on PGPR gene expression and plant beneficial properties was thus studied by performing (i) the chemical characterization of a new Azospirillum plant-stimulatory molecule whose positive impact on root development was revealed, (ii) an approach targeting the in situ expression of the acdS plant beneficial gene in PGPR, and (iii) a global approach to identify the Azospirillum transcriptomic response to a bacterial secondary metabolite. This works allowed the identification of rhizospheric signals controlling PGPR root colonization and/or plant growth promotion
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Effet de l’exsudation de citrate chez le lupin blanc sur la phytodisponibilité de l’uranium / Effect of citrate exudation in white lupine on uranium phytoavailabilityTailliez, Antoine 08 July 2013 (has links)
L’étude proposée s’inscrit dans un contexte de contamination croissante de la pédosphère et fait le choix d’appréhender cette problématique via une approche centrée sur la rhizospère et sur les mécanismes gouvernant son évolution. Ces travaux s'intéressent à la relation entre l’uranium – un métal naturel ubiquiste non-essentiel – et le citrate – un acide organique de faible poids moléculaire exsudé par certaines espèces végétales poussant sur un milieu pauvre en phosphore. Parmi ces espèces particulières le lupin blanc (Lupinus albus) fait office de plante modèle. Un sol naturellement riche en uranium prélevé à l’affleurement d’une veine de pechblende a été étudié. L’objectif du travail a été de savoir si l’activité racinaire est suffisante pour provoquer la remobilisation de l’uranium dans la solution du sol et favoriser son transfert. Le système sol/plante a été étudié sur rhizotest et de manière découplée en hydroponie et en réacteurs fermés. Il a été démontré que la présence d’uranium dans un milieu contrôlé (hydroponie) stimule l’exsudation de citrate chez les lupins blancs qui ne sont pas carencés en phosphore et que cette exsudation de citrate favorise la solubilisation de l’uranium dans la solution du sol en réacteurs fermés. Ces résultats ont été confirmés par les observations faites sur rhizotest même si les valeurs obtenues dans ce cas étaient plus nuancées. Ceci justifie les approches utilisées et met en évidence leur complémentarité. Les résultats obtenus ont également permis d’apporter des informations concernant la toxicité de l’uranium, son internalisation dans les cellules racinaires ainsi que sur son transfert des racines aux parties aériennes. / The present study is related to the context of pedosphere contamination increasing and chose to consider this issue through a focus on rhizosphere and mechanisms that govern its evolution. This work is oriented on the relation between uranium – a non-essential ubiquitous metal – and citrate – a low molecular weight organic acid exudated by specific species when grown on a phosphorus-deficient environment. Among these particular species white lupine (Lupinus albus) is considered as a model species. Furthermore, a natural soil containing huge amounts of uranium has been collected at the outcrop of a pitchblende vein and used during some experiments. The main aim of this PhD thesis was to determine if the root activity of white lupine is sufficient to cause the remobilization of uranium into soil solution and so to promote the transfer of this contaminant to the plant. The soil/plant system has been studied with rhizotests and by combining datas obtained with hydroponics experiments and batches tests. It has been demonstrated that the presence of uranium in a controlled growth medium (hydropony) stimulates citrate exudation in P-sufficient white lupine. More, this citrate exudation enhances uranium solubilization in soil solution in batches. These results have been confirmed by observations on rhizotest even if the values in this case were more nuanced. This justifies the employed approaches and highlights their coherence and complementarity. The results also allowed to provide additional information concerning uranium toxicity, its internalization in root cells as well as its transfer from roots to aerial parts.
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Fatores determinantes na composição da comunidade bacteriana associada às plantas / Determinant factors in the composition of bacterial communities with plants associatedAndreote, Fernando Dini 08 October 2007 (has links)
A interação entre bactérias e plantas resulta na ocorrência de vários processos biológicos no ambiente e pode ser regulada por diferentes fatores. Considerando que um recíproco reconhecimento ocorre entre as espécies de bactérias e plantas, alterações nos fatores bióticos e abióticos interferem diretamente nesta interação levando a modificações na composição das comunidades bacterianas associadas às plantas. No presente trabalho foram avaliados diferentes fatores que estão diretamente relacionados a este assunto. Atualmente, a aplicação de técnicas de microbiologia molecular permite acessar alterações causadas nestas comunidades de maneira independente do cultivo bacteriano. Desta maneira, foi demonstrado que na rizosfera de plantas de tabaco, os diferentes estágios de desenvolvimento são os principais determinantes da composição da comunidade bacteriana em detrimento do genótipo das plantas, sejam estas transgênicas ou convencionais. Utilizando plantas transgênicas e convencionais de eucalipto de diferentes genótipos, foi verificado que diferentes plantas não transgênicas podem apresentar comunidades bacterianas mais distintas do que quando os clones não transgênicos são comparados com os transgênicos. No entanto, efeitos específicos podem ocorrer na interação bactéria-planta, como o observado pela inibição da população de Methylobacterium spp. em plantas transgênicas de eucalipto TR-15. Avaliando o efeito da inoculação de bactérias endofíticas na composição de comunidades bacterianas associadas à plantas de batata de diferentes cultivares, os resultados mostram que a inoculação de Pseudomonas putida altera a comunidade bacteriana de maneira similar a alteração da variedade da planta. Os demais isolados avaliados, classificados como Paenibacillus sp. e M. mesophilicum, causam menores alterações na composição das comunidades bacterianas. Adicionalmente foi demonstrado que a colonização das plantas de batata pelo endófito P. putida resulta em pequena alteração no perfil metabólico da planta hospedeira. Por fim, buscando melhores métodos de isolamento da comunidade bacteriana da rizosfera de batata, os resultados mostraram que a mimetização do ambiente pode resultar em melhor acesso da diversidade bacteriana por meio de isolamento e cultivo das espécies componentes desta comunidade. / The plant-bacteria interactions result in the occurrence of biological process in the environment and might be regulated by different factors. Considering that a reciprocal recognition occur amongst bacteria and plants species, shifts in biotic and abiotic factors interfere directly on these interactions, leading to modifications in the composition of bacterial communities to plants associated. In the present work different factors related to this subject were evaluated. Nowadays, the applications of techniques of molecular microbiology allow assessing the shifts caused on these communities by a culture independent approach. On this way, it was demonstrated that in rhizosphere of tobacco plants, different stages of plants development are main determinants of bacterial community composition rather than the plants genotypes, transgenic or not. Using plants transgenic or not, carrying different genotypes, it was verified that different non-transgenic plants could harbor bacterial communities more distinct than those observed in association with transgenic plants. However, specific effects could be observed like the inhibition of Methylobacterium spp. population in eucalyptus transgenic plants TR-15. Considering the effect of endophytic bacteria inoculation in the composition of bacterial communities associated to different cultivars of potato plants, the results show that inoculation of Pseudomonas putida causes similar shifts in the bacterial community similarly to those observed when different cultivars are considered. Other evaluated strains, classified as Paenibacillus sp. and M. mesophilicum, caused minor alterations in the composition of bacterial communities. In addition, it was demonstrated that plant colonization by endophytic P. putida results in small effects on the metabolic profile of host plant. At least, aiming better methodologies for bacteria isolation from potato rhizosphere, the results show that mimicking the natural environment could result in a better assessment of bacterial diversity by isolation of species present in this community.
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Actinobactérias da rizosfera de Araucaria angustifolia com potencial biotecnológico / Rhizosphere Actinobacteria from Araucaria angustifolia with biotechnological potentialVasconcellos, Rafael Leandro de Figueiredo 31 October 2008 (has links)
A espécie Araucaria angustifolia, pertencente ao fragilizado Bioma Mata Atlântica, foi, durante décadas, uma das maiores fontes de madeira no Brasil. Essa espécie é de grande importância sendo fonte de alimento e matéria prima para a produção de móveis, polpa de celulose e vernizes. Devido à grande importância econômica e ambiental da araucária, estudos voltados à preservação e manejo da espécie se tornaram necessários. Este trabalho teve como finalidade isolar actinobactérias, presentes na rizosfera de araucária, antagônicos aos fungos Fusarium sp .e Armillaria sp. Estes patógenos são responsáveis por danos às raízes e sementes causando podridão e perda de mudas. Além disso, verificou-se também o efeito das actinobactérias sobre a germinação de esporos de Gigaspora rosea. Após a seleção dos melhores inibidores, identificação morfológica e seqüenciamento do gene 16S rRNA, verificou-se o efeito destes sobre o crescimento de Pinus taeda, na presença e na ausência do fungo ectomicorrízico (Suillus brevipes). Os isolados ainda foram avaliados quanto à produção de enzimas como quitinase, lipase, fosfatase, celulase, amilase e protease. Para o isolamento foram coletadas raízes de 15 árvores adultas presentes na mata nativa. Dez gramas de raízes frescas com resíduos de solo aderidos firmemente foram agitados por 30 minutos em solução salina 0,85 % padronizando como solo rizosférico aquele que soltou das raízes. Neste isolamento foram utilizadas duas metodologias: por plaqueamento e por separação física pela utilização de membrana de 0,45 micrômetros. Foram obtidos 33 possíveis actinobactérias. Os isolados foram testados quanto à capacidade antagônica pelo método de contato direto, inoculando-os a 30 mm de distância do fungo fitopatogênico Fusarium sp., em placas de petri contendo meio ISP2. Os halos de inibição foram medidos após 3, 5, 7 e 10 dias. De todos os isolados testados 6 mantiveram a inibição por 10 dias com manutenção de halos de inibição de até 4 mm contra um Fusarium sp. isolado de sementes de milho e 2 foram eficientes na inibição de Fusarium sp, patógeno de Pinus sp. A análise de inibição do fungo Armillaria sp, foi feito em meio líquido medindo o crescimento em mg/dia após 30 dias na presença de extratos de actinobactérias e também pela contagem de rizomorfas em placa de petri após 20 dias de incubação em contato direto com as actinobactérias. Verificou-se que de 28 isolados testados 24 foram capazes de inibir a produção de rizomorfas, destacando-se o isolado A43 que foi capaz de inibir ambos os fungos, Fusarium e Armillaria. Os esporos de Gigaspora rosea tiveram a taxa de germinação avaliada na presença de actinobactérias a partir da técnica de dupla camada. Todos os seis isolados testados foram capazes de estimular a germinação dos esporos desse fungo micorrizico. Nenhum dos seis isolados (A43, A43b, PNA, A64, A75 e A93) foi capaz de produzir fosfatases e lipases. Porém os isolados A93, A75, A64 e PNA produziram protease, quitinase e amilase. Em casa de vegetação, foi testado o efeito de seis isolados, na presença e não ausência de ectomicorriza (Suillus brevipes), sobre plântulas de Pinus taeda. Analisou-se o diâmetro, a altura, a massa seca da raiz e da parte aérea e também o fósforo da parte aérea. Destaca-se o isolado A43 que, quando na ausência de ectomicorriza, estimulou o desenvolvimento da planta em relação ao controletambém sem ectomicorriza, provocando aumento de 100 % para massa seca da parte aérea e da raiz. Os resultados encontrados neste trabalho poderão levar ao desenvolvimento de novas tecnologias, visando à identificação de novos metabólitos e novas técnicas de manejo, voltados ao controle de doenças de plantas, especialmente as espécies arbóreas. / The tree Araucaria angustifolia, belonging to the endangered Atlantic Forest biome, for many decades was the source of Brazilian wood. This species is also very important in providing food and feed, as well as raw material for joinery, cellulose pulp and varnish. Due to the economic and environmental importance of A. angustifolia, research projects involving the preservation and management of this species are becoming more urgent and necessary. The aim of this work was to isolate Araucaria rhizosphere actinobacteria with antagonic effects against the plant pathogens Fusarium sp. and Armillaria sp. These fungi cause root rot and seed damage, with the consequent loss of seedlings. Moreover, the effect of these actinobacteria on Gigaspora rosea spore germination was studied. After the selection of the best pathogen inhibitors, we also tested the effect of these microorganisms on Pinus taeda growth, in the presence or absence of the ectomycorrhizal fungus Suillus brevipes. The production of protease, chitinase, lipase, phosphatase, cellulase and amylase of these bacteria in culture media was also investigated. For the isolation of rhizosphere bacteria, we collected roots of 15 adult trees in a native forest. Ten grams of fresh roots with soil residues adhered to the surface were shaken in 0,85 % salt solution for 30 minutes. Two techniques were used, the dilution plate method and the coverage of the medium, utilizing a 0,45 µm membrane to separate these filamentous bacteria. About 33 actinobacteria were isolated. After isolation the actinobacteria were tested against the plant pathogenic fungi Fusarium spp., utilizing dual culture techniques and ISP2 medium. The inhibition halo was measured after 3, 5, 7 and 10 days. Six of our isolates maintained an inhibition zone measuring at least 4 mm against Fusarium sp. isolated from corn seed and 2 mm against the Fusarium, which causes root root of Pine trees. For the inhibition test of Armillaria, in liquid medium with the addition of culture extracts of actinobacteria, the growth in mg/day was measured after thirty days growth, and the number of rizomorphs produced in culture dishes after twenty days in dual culture with the actinobacteria was counted. Six bacteria proved to be antagonistic (A43, A43b, A64, PNA, A93 e A75), and only one had no effect. Possibly the elevation of the pH value played also a role in this situation. About 24 of 28 isolates inhibited the rizomorph production, especially the isolate A43 that showed a double antagonism against Fusarium and Armillaria. The dual layer test was used to investigate the reaction of the arbuscular mycorrhizal fungus Gigaspora rosea spore germination to the presence of actinobacteria. All the six actinobacteria stimulated the germination, but the germ tube did not grow straight forward as in the control. This result may indicate a negative effect against this arbuscular mycorrhizal fungus. None of the six isolates tested (A43, A43b, PNA, A64, A75 and A93) produced phosphatases and lipases, but A93, A75, A64 and PNA produced protease, amylase and chitinase. Isolates A43 and A43b did not produce any of the enzymes tested. This fact suggests that there is production of an antibiotic acting against the pathogenic fungi. Pinus taeda seedlings were grown under green-house conditions. After three months the stem diameter, shoot height, root and shoot dry weight and shoot phosphorus content were evaluated. Plants with ectomycorrhiza presented a significant growth promotion in comparison with the nonmycorrhizal ones. Among the actinobacteria in the absence of mycorrhiza only the isolate A43 produced a 100% growth enhancement in comparison with the control plant without ectomycorrhiza. The results presented in this dissertation could lead to the development of new technologies and new management techniques, with regard to the control of plant diseases, especially in tree species.
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Untersuchungen zum Einfluß selektierter arbuskulärer Mykorrhizapilze (AMP) und assoziativer Rhizosphärenbakterien einzeln und kombiniert auf das Wachstum und den Zierwert von Zierpflanzen für den urbanen BereichJahn, Mareile 01 January 1998 (has links)
In mehrjährigen Gefäß- und Feldversuchen wurde der Einfluß von selektierten arbuskulären Mykorrhizapilzen (Glomus ssp. VAM3, Glomus intraradices Isolat 49) und assoziativen Rhizosphärenbakterien (Pseudomonas fluorescens PsIA12, Agrobacterium rhizogenes A1A4, Rhizobium trifolii R39, Stenotrophomas maltophilia PsIB2 und PsI2) einzeln und kombiniert auf das Wachstum und den Zierwert von ein- und mehrjährigen Zierpflanzen für den urbanen Bereich untersucht. Auf urbanen streßbelasteten Standorten führten diese Rhizosphärenmikroorganismen ohne zusätzliche Mineraldüngung zur Förderung von Wachstum und Zierwert bei Zierpflanzen. Frühzeitige Inokulationen zur Aussaat bzw. Pflanzung verkürzten die Pflanzenanzucht und reduzierten Pflanzenausfälle. Die drei Pflanzenarten reagierten unterschiedlich. Die deutlichsten Effekte wurden bei Tagetes durch Einzelinokulationen erzielt. Es zeichneten sich sortenspezifische Reaktionen ab. Bei Miscanthus waren Kombinationen von Mikroorganismen wirksam und förderten das Wachstum über drei Jahre. Nicht eindeutig reproduzierbar waren die Wirkungen bei Gladiolen, der Zierwert wurde positiv beeinflußt. Alle Bakterienstämme produzierten in Reinkultur Auxine und z.T. Cytokinine. Zwischen Phytohormonbildung, Mykorrhizierung der Wurzeln, Wurzelbesiedlung durch die autochthone Mikroflora und Wurzelstimulierung zeichneten sich wiederholt positive Wechselwirkungen ab. Von Ackerstandorten isolierte Bakterien waren auch auf urbanen nährstoffarmen Standorten wirksam. Die Bakterien besiedelten die Rhizosphäre von Tagetes und Gladiolen während der Vegetationsperiode im Freiland. Nach einjähriger Trockenlagerung des Bodens besiedelten PsIA12 und PsIB2 erneut die Rhizosphäre der nichtinokulierten Tagetes. / In long-term pot and field trials the influence of selected arbuscular mycorrhizal fungi (Glomus ssp. VAM3, Glomus intraradices Isolat 49) and associative rhizosphere bacteria (Pseudomonas fluorescens PsIA12, Agrobacterium rhizogenes A1A4, Rhizobium trifolii R39, Stenotrophomas maltophilia PsIB2 and PsI2), single and in combination, on growth and ornamental value of annual and perennial ornamental plants under urban conditions was tested. These rhizosphere microorganisms did stimulate growth and ornamental value of ornamental plants without the additon of mineral fertilizers on urban stressed sites. An early inoculation during seeding or planting did shorten plant cultivation and reduced plant losses. The three plant species did respond differently. The most obvious effects were achieved by single inoculation on Tagetes. Variety specific reactions did occur. Combination of microorganisms was effective on Miscanthus and did stimulate growth over three years. The effects on Gladiolus were not repeatable but ornamental value was stimulated. All bacteria strains did produce Auxin and partially Cytokinin in pure culture. Between phytohormone production, root mycorrhization and root colonization by the autochthonous microflora positive interactions did occur. Bacteria isolated from agricultural sites were also effective on oligotrophic urban sites. The bacteria did establish in the rhizosphere of Tagetes and Gladiolus over the vegetation period on the field. After one year dry storage of the soil PsIA12 and PsIB2 did re-establish in the non-inoculated Tagetes rhizosphere.
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Weathering and incipient pedogenesis of Technosols constructed from dolomitic limestone mine spoils / Intemperismo e pedogênese incipiente em Tecnossolos desenvolvidos a partir de rejeitos de mineração de calcário dolomíticoRuiz, Francisco 07 February 2019 (has links)
Technosols combine soils subjected to strong human influence and whose pedogenesis is dominated by its technical origin. They contain a significant amount of materials made, modified or directly transported by human activity, defined as artefacts. Soils formed from technogenic parent materials are spreading and their pedogenesis is still little understood. Given that the sustainable use of the soil resources demands wide knowledge about its genesis, it is necessary to bring further information about Technosols, especially in the tropical environment, where they have been poorly explored. Thus, in this study we characterized three Technosols constructed from dolomitic limestone mine spoils, cultivated with sugar cane (for 2 and 6 years) and pasture (for 20 years). The Technosols experienced a rapid morphological evolution expressed by color development and aggregation. Pedoplasmation, melanization and redoximorphic were the mainly processes occurring in these constructed soils. The combination of grasses over Ca-rich mine spoils triggered organic carbon accumulation and originated a well-developed surface horizon with strong crumb structure and dark color. The rhizosphere played a pivotal role in mineral weathering of the Technosols. Exudation and microbial activity were about two times higher in the rhizosphere than in the bulk soil leading to an enhanced mineral dissolution. Overall, the primary minerals in the rhizosphere presented a medium to high degree of weathering while those from bulk soil were little weathered. Our data indicates that the rhizosphere may provide a favorable biogeochemical environment to the formation of secondary clay minerals. This study evidences that constructed tropical soils may evolve in an accelerated rate of pedogenesis and, thus, are a real alternative for the reclamation of degraded areas, potentially restoring original soil ecosystem services. / Tecnossolos são uma classe de solos cuja pedogênese é dominada por sua origem antrópica. Esses solos contêm uma quantidade significativa de materiais feitos, modificados ou transportados diretamente pela atividade humana, definidos como artefatos. Solos formados a partir de materiais de origem antrópicos são muito comuns, entretanto sua pedogênese ainda é pouco compreendida. O uso sustentável do recurso solo exige um amplo conhecimento sobre sua gênese, sendo assim, entender a formação de Tecnossolos, especialmente no ambiente tropicalé crucial para seu uso e manejo visando sua incorporação na produçao de alimento, fibras e energia. Neste estudo foram estudados três Tecnossolos construídos a partir de rejeitos de mineração de calcário dolomítico, cultivados por dois e seis anos com cana-de-acúcar e por 20 anos sob pastagem. Os Tecnossolos apresentaram uma rápida evolução morfológica expressa principalmente pelo desenvolvimento de cor e agregação. Dentre os processos identificados, os mais importantes foram pedoplasmação, melanização e os redoximórficos. Devido à combinação de gramíneas e rejeitos ricos em cálcio desenvolveu-se um horizonte de superfície com estrutura grumosa forte e cor escura, com altos teores de carbono orgânico. A rizosfera desempenhou um papel fundamental no intemperismo mineral dos Tecnossolos. A exsudação e a atividade microbiana foram cerca de duas vezes maiores na rizosfera do que no solo não rizosférico, levando a uma dissolução mineral mais proeminente na rizosfera. No geral, os minerais primários da rizosfera apresentaram um grau médio/alto de intemperismo, enquanto aqueles do solo não rizosférico foram pouco intemperizados. As condições biogeoquímicas da rizosfera apontam para um ambiente favorável à formação de minerais de argila na rizosfera. O estudo demonstra que a pedogênese de Tecnossolos emambiente tropical parece avançar rapidamente; o que pode garantir não só o sucesso da reabilitação das áreas, mas ainda a retomada de serviços ecossistêmicos prestados pelos solos.
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Phosphorus dynamics in the rhizosphere of sugarcane under phosphate sources and filter cake / Dinâmica do fósforo na rizosfera de cana-de-açúcar sob fontes fosfatadas e torta de filtroArruda, Bruna 11 November 2015 (has links)
In tropical soils, phosphorus (P) is adsorbed onto the oxide surface reducing the availability to plants. Thus, the solubility of the P sources and the particular plant characteristics which increases the absorption efficiency becomes important in the rhizosphere where roots and soil microorganisms interacts together. The objectives of this research were to evaluate sugarcane cultivars in relation to changes in the rhizosphere and the efficiency of P absorption and to evaluate the dynamics of P in the rhizosphere of sugarcane under the application of filter cake combined with mineral fertilizers. Four experiments were conducted in a greenhouse. The soil was a sandy clay loam Ferralsol from Piracicaba - SP, with low soil P, arranged in a randomized block design with four replications for each experiment. The experiment 1 was conducted in a 5x2 factorial arrangement with four sugarcane cultivars: RB92-579; RB85-5156; RB86-7515; RB86-6928 or no plants subjected to presence or absence of phosphate fertilizer (78.4 mg P kg-1) and soil adhering to the root was considered rhizosphere. The experiment 2 evaluated five doses of phosphate fertilizers: 0; 9.8; 19.6; 39.2 and 78.4 mg P kg-1 soil with the most promising cultivar for P absorption from experiment 1, and the rhizosphere was collected in five distances from the rhizoplane (mm): i) 0-2; ii) 2-4; iii) 4-6; iv) 6-8; v) 8-10. Experiment 3 was conducted in a 2x3 factorial: absence or presence of filter cake (5 g DM kg-1) with rock phosphate, soluble phosphate (78.4 mg soluble P kg-1 soil) or phosphate absence, and the rhizosphere soil was sampled 2 mm from the rhizoplane. Experiment 4 involved five filter cake rates: 0; 2.5; 5; 10 and 15 g DM kg-1 applied to the bulk soil or in the planting furrow. The cultivar RB96-6928 showed better adaptation characteristics under low availability of phosphorus, with good root development and P uptake. The rate 38.5 mg P kg-1 was the most suitable to promote optimal seedling development. The filter cake was efficient to increase P uptake and the interaction between filter cake and rock phosphate was efficient to increase production factors as the number of tillers per plant. The highest P uptake occurred when filter cake was applied to planting furrow at a rate of 10 g DM kg-1. / Em solos tropicais o fósforo (P) é adsorvido nas superfícies dos óxidos, reduzindo a disponibilidade às plantas. Assim, a solubilidade das fontes de P e as características particulares da planta que aumentam a eficiência de absorção tornam-se importantes na rizosfera, onde as raízes, microrganismos e solo interagem biologicamente de forma intensa. Os objetivos desta pesquisa foram avaliar cultivares de cana-de-açúcar quanto a mudanças na rizosfera e eficiência na absorção de P e avaliar a dinâmica do P na rizosfera de cana-de-açúcar sob a aplicação de torta de filtro juntamente com fertilizantes minerais. Foram conduzidos quatro experimentos em casa de vegetação. O solo utilizado foi um Latossolo Vermelho amarelo arenoso da região de Piracicaba - SP, com baixo teor de P, em delineamento em blocos ao acaso com quatro repetições para todos os experimentos. O experimento 1 foi conduzido em esquema fatorial 5x2, com 4 cultivares de cana-de-açúcar: RB92-579; RB85-5156; RB86-7515; RB86-6928 ou ausência de plantas, submetidos a presença ou ausência de adubação fosfatada (78.4 mg P kg-1), e o solo aderido a raiz foi amostrado com sendo rizosférico. O experimento 2 avaliou cinco doses de adubação fosfatada: 0; 9.8; 19.6; 39.2 e 78.4 mg P kg-1 solo com o cultivar mais promissor para absorção de P do experimento 1, sendo o solo rizosférico coletado em cinco distâncias a partir do rizoplano (mm): i) 0-2; ii) 2-4; iii) 4-6; iv) 6-8; v) 8-10. O experimento 3 foi conduzido em esquema fatorial 2x3: ausência ou presença de torta de filtro (5 g MS kg-1) com fosfato natural, fosfato solúvel (78.4 mg P solúvel kg-1 solo) ou ausência de fosfato, sendo o solo rizosférico amostrado a 2 mm do rizoplano. O experimento 4 envolveu cinco doses de torta de filtro: 0; 2.5; 5; 10 e 15 g MS kg-1 aplicados no volume total do solo ou no sulco de plantio. A cultivar RB96-6928 mostrou características de melhor adaptação sob baixa disponibilidade de P, com destacado desenvolvimento radicular e boa absorção de P. A dose 38.5 mg P kg-1 foi a mais apropriada para promover um adequado desenvolvimento das mudas. A torta de filtro foi eficiente para aumentar a absorção de P e a interação entre torta de filtro e fosfato natural foi eficiente para incrementar fatores de produção como o número de perfilhos por planta. A maior absorção de P se deu quando a torta de filtro foi aplicada no sulco de plantio na dose de 10 g MS kg-1.
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