Ce travail a comme objectif d’explorer le potentiel de bioremédiation de plantes et de rhizobactéries pour la réhabilitation des sols de sites miniers dans la région de Marrakech. Nous avons isolé des souches rhizosphériques et nodulaires des racines de fève et de plantes indigènes des sites miniers Draa Sfar et Kettara. Plusieurs isolats ont montré une bonne tolérance aux ETM allant jusqu’à 25 mg/l de Cu, 200 mg/l de Pb et 150 mg/l de Zn. Les souches tolérantes aux métaux ont fait l’objet de criblage pour déterminer leurs caractères PGPR (production d’exopolysaccharides, sidérophores, AIA et activité antifongique). En présence du sol minier amendé avec le sol agricole, la fève a montré une capacité à accumuler les métaux différemment d’un métal à l’autre. Le Pb a été peu mobile dans les parties aériennes des plantes, et n’a pas atteint les graines. Le Zn a été le métal le plus transloqué vers les parties aériennes bien que sa quantité soit importante au niveau des racines. Le Cu a été accumulé plutôt dans les racines des plantes. Le Zn et le Cu ont été transloqués aux graines mais à des teneurs significativement faibles. L’inoculation par les bactéries PGPR a permis de mieux accumuler le Cu dans les plantes notamment dans la partie racinaire. L’inoculation mixte PGPR-rhizobia a amélioré l’absorption du Zn dans les plantes notamment dans la partie aérienne. Les analyses in natura sur ces sites miniers fortement contaminés par des métaux lourds indiquent que le microbiote a évolué, devenant tolérant à la pollution à long terme par les métaux lourds et adapté aux plantes indigènes elles-mêmes capables de tolérer et d'accumuler les métaux lourds. / This work aims to explore the potential of plant and rhizobacteria for bioremediation in mining soil in Marrakech region. Rhizospheric and nodules strains were isolated from faba bean roots and native plants growing in Draa Sfar and Kettara mining sites. Several isolates showed good tolerance to metals up to 25 mg/l of Cu, 200 mg/l of Pb and 150 mg/l of Zn. The tolerant strains to metals were screened for their PGPR characteristics (production of exopolysaccharides, siderophores, IAA and antifungal activity). In mining soil amended with agricultural soil, faba bean showed an ability to accumulate metals differently. Pb was not very mobile in shoots, and did not reach the seeds. Zn was the most translocated metal to the aerial parts although its amount was important in roots. Cu was accumulated rather in the roots of plants than shoots. Zn and Cu were translocated to seeds but at significantly lower levels. The inoculation with PGPR bacteria allowed Cu to accumulate better in plants, particularly in roots. Whereas the PGPR-rhizobia mixed inoculation improved the absorption of Zn in plants, especially in shoots. In natura investigation of mining sites heavily contaminated with heavy metals (HMs) indicates that microbiota has evolved, becoming tolerant to long-term HMs pollution and adapted to indigenous plants themselves able of tolerating and accumulating HMs.

Der Einfluss der Rhizosphärenbakterien Bacillus subtilis FZB 24, FZB 37, FZB 42, Bacillus pumilus RK 13 und Streptomyces graminofaciens und des Algenpräparates Goemar Fruton Spezial® (Ascophyllum nodosum) auf Wachstum, Ertrag und Toleranz gegenüber Fusarium spp. bei Spargel wurde untersucht. Es wurden mehrjährige Parzellenfeldversuche und Modellversuche unter kontrollierten Bedingungen mit natürlich Fusarium spp. belasteten Böden und mit speziellen Fusarium Erregern inokulierten Substraten durchgeführt. Die Analyse des verwendeten Nachbaubodens ergab eine wesentlich höhere Kontamination mit Fusarium spp. (56%) in Relation zum Fruchtfolgeboden (14%). F. oxysporum war die dominante Fusariumart. Desweiteren wurde eine ganze Reihe anderer Arten, wie z.B. F. acuminatum, F. culmorum, F. proliferatum und F. culmorum bestimmt, was die Komplexität der Wurzel- und Stängelfäuleerkrankung belegt. Analog zu diesem Ergebnis konnten im Vergleich zur Fruchtfolgevariante von den in dem Nachbauboden kultivierten Spargelpflanzen signifikant mehr Fusarium spp. (80%) isoliert werden. Das Wurzelwachstum war hier extrem reduziert. Unter diesen konduktiven Bedingungen gelang es durch Rhizombakterisierung (107 cfu/ml) mit B. subtilis FZB 42, eine signifikante Förderung des Wurzelwachstums relativ zur unbehandelten Kontrolle und zur B. subtilis FZB 37 Variante zu erzielen. In Pathogenitätstests konnten F. culmorum, F. oxysporum und F. proliferatum als bedeutende Spargelpathogene mit signifikant reduzierter Trieb- und Wurzelmasse nachgewiesen werden, während F. acuminatum nur geringe Symptome verursachte. Eine bakterielle Saatgutbeizung (108 cfu/ml) und zusätzliche präinfektionelle Gießapplikation (107 cfu/ml) konnte eine Infektion mit F. oxysporum f.sp. asparagi nicht verhindern. Dennoch waren die negativen Effekte im Vergleich mit der nicht bakterisierten Kontrolle in den B. subtilis Varianten FZB 24 und vor allem bei FZB 42 deutlich kompensiert. Diese Ergebnisse lassen vermuten, dass eine Resistenz- bzw. Toleranzinduktion ein potentieller Wirkmechanismus der Bakterien ist. In den Feldversuchen erbrachte eine Bakterisierung einjähriger Rhizome (107 cfu/ml) zur Pflanzung und ergänzende Gießbehandlungen (108 cfu/ml/1l/m) in den folgenden 2 Jahren eine tendenzielle Reduzierung der Trieblänge und des Triebdurchmessers, insbesondere in Kombination mit dem Algenpräparat. Einen eindeutigen Einfluss auf das Sortierungsergebnis konnte nicht nachgewiesen werden. Im Gegensatz hierzu resultierte eine Saatgutbakterisierung mit B. subtilis FZB 24 in einem generell geförderten Triebwachstum und einer signifikant gesteigerten Wurzelentwicklung. Auch die anderen Mikroorganismen erbrachten eine tendenzielle Wuchsförderung. Es konnte eine gesicherte Korrelation zwischen Wurzelmasse und Knospenanzahl ermittelt werden. Blattapplikationen mit Goemar Fruton Spezial® während der Hauptvegetationsphase hatten keinen Einfluss auf das Pflanzenwachstum. / The effects of the rhizobacteria Bacillus subtilis, Strain FZB 24, FZB 37and FZB 42, Bacillus pumilus RK 13, Streptomyces graminofaciens N6 and the alga Ascophyllum nodosum (Goemar Fruton Spezial®) on plant growth, yield and Fusarium spp. tolerance of Asparagus officinalis (L.) were investigated. The trials were carried out under field conditions over sev-eral years by sawing and planting and as pot trials in the greenhouse and climate chamber with soil naturally infested with Fusarium spp. and with steamed substrates inoculated with particularly Fusarium species. Asparagus replant soil was considerable higher infested with Fusarium spp. (56 %) than fresh soil without asparagus history (14 %). F. oxysporum was the most determined Fusarium species followed by F. redolens, F. acuminatum, F. culmorum, F. proliferatum, F. solani a.o.. That indicates the complex character of root and crown rot of asparagus. From asparagus plants cultivated in replant soil a significant higher rate of Fusarium isolations (80 %) could be worked out compared to the fresh soil treatment. The root growth in the contaminated soil was extremely reduced. In these conductive situation bacterial treatments with B. subtilis FZB 42 done as rhizom soaking (107cfu/ml) resulted in significantly increased root growth up to 32, 9 % related to the non treated control. B. subtilis FZB 37 was ineffective. F. oxysporum, F. culmorum and F. proliferatum showed a high pathogenicity to asparagus seedlings, manifested in significantly reduced fresh and dry weights of shoots and roots, while F. acuminatum caused only slight symptoms. None of the tested microorganisms applicated as seed coating (108 cfu/ml) and substrate drenching before pathogen inoculation (107 cfu/ml) could prevent seedlings from being infected by F. oxysporum f. sp. asparagi. The negative effects of infection are significantly compensated by Bacillus subtilis FZB 42 and FZB 24. The results support the conclusion, that induced tolerance is a potential mechanism of bacterial mode of action. Under field conditions rhizom bacterization of one year old asparagus plants (107 cfu/ml) before planting and additional soil drenching (108 cfu/ml/1,5l/m) during the following two years reduced shoot length, shoot diameter and yield, especially in combination with Goemar Fruton Spezial®. There was no remarkable influence on spear quality. Seed coating with B. subtilis FZB 24 (108 cfu/ml) and a soil drenching (108 cfu/ml/1l/m) in summer however re-sulted in higher shoots and significantly increased fresh weight and dry substance of roots. But also the other tested microorganisms showed a plant growth promoting trend. There was a significant positive correlation between root fresh weight and number of buds. By spraying the phylloclads four times during the main vegetation season the alga application only gave a slight improvement of plant growth in the field trials.

Resumo: O cultivo intensivo agrícola, apesar de altamente produtivo, possui diversos pontos questionáveis, como suplementação acentuada de insumos sintéticos, além da aplicação de defensivos para o controle de pragas que muitas vezes são danosos ao meio ambiente. Como alternativa, a agricultura orgânica propõe uma forma de manejo menos agressiva, que promove o uso sustentável dos recursos naturais renováveis, como o aproveitamento de resíduos orgânicos, além de possuir um maior valor agregado no produto final. No presente estudo, investigamos por meio de uma abordagem metagenômica os efeitos do manejo (orgânico ou convencional) na microbiota da rizosfera de cana-de-açúcar, região da planta que notoriamente causa um grande impacto no seu desenvolvimento e produtividade. Identificamos a abundância maior na rizosfera com manejo orgânico de porções genômicas que codificam enzimas associadas ao metabolismo do nitrogênio e enxofre, sendo que estes são nutrientes fundamentais para o sucesso da planta e assim, seu desenvolvimento e produtividade em campo. Contraditoriamente, encontramos também que os genomas da microbiota orgânica possuem conteúdo ligado a um potencial mais acentuado para a degradação de compostos xenobióticos, que são ativamente aplicados no manejo convencional. De acordo com nossa predição, os microrganismos dessa microbiota poderiam realizar sua completa mineralização, o que é muito favorável na remoção desses resíduos no ambiente. Esses achados indicam que há um potencia... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Modern industrialized agriculture although highly productive, has several sustainability problems such as the for intense supplementation of synthetic inputs, beyond the application of pesticides for pest control. Which are often harmful to environment. As an alternative to this system, organic agriculture is considered more environmentally progressive, having several characteristics favorable to the crop producer, promotes sustainable use of renewable natural resources, such as the use of organic waste, besides that a higher aggregated value in the final product. This study investigates the changes that an organic system brings when compared to conventional management for the sugarcane rhizosphere microbiome, a plant region known to be determinant of plant development and associated with net productivity, using a metagenomic approach. We identified a higher abundance of genomic portions which encode enzymes associated with nitrogen and sulphur metabolism, which are fundamental nutrients for plant success and subsequently, crop productivity. Conversely, we also found that genomes of the organic microbiome contain higher portions linked to xenobiotic degradation, compounds that are actively applied in conventional management. Microorganisms from this sequenced microbiome could perform complete mineralization of these damaging compounds, which is desirable in their removal from the environment. These findings indicate that there is a unique potential for exploitation of organic... (Complete abstract click electronic access below) / Mestre

Inoculation with free-living diazotrophic bacteria is well documented to enhance vegetative growth and yield increases of various crops coupled with suppression of sublethal pathogens. The use of microbial inoculants has been identified as an alternative or supplement to use of nitrogenous fertilizers and agrochemicals for sustainable agriculture. The search for effective free-living diazotrophic bacterial strains for formulation as biofertilizers has been on going since the 1970’s and a number of inoculant biofertilizers have been developed and are commercially available. In the current study, 250 free-living diazotrophic bacteria were isolated from soils collected from the rhizosphere and leaves of different crops in different areas within KwaZulu-Natal, province, Republic of South Africa. These were evaluated for plant growth-promotion and biological control of damping-off fungi initially by in vitro screening. The growth promotion traits tested included, phosphate-solubilization, production of indole-3-acetic acid, production of ammonia and acetylene reduction. Biocontrol traits evaluated included siderophore-production, antibiosis, and production of hydrogen cyanide (HCN). Biochemical and molecular bioassay tests were conducted to identify the twenty most promising isolates selected in the in vitro study. The twenty isolates were further tested in combination with various levels of nitrogenous fertilizer for growth-promotion of maize (Zea mays L.) and wheat (Triticum aestivum L.) under greenhouse conditions. The five most promising isolates identified for growth promotion under greenhouse conditions for each crop were assessed for their effects on the germination of maize in vitro and growth and yields of two maize and two wheat cultivars, when combined with a low dose of nitrogenous fertilizer in field trials. The five Bacillus subtilis (Ehrenberg) Cohn isolates that suppressed the growth of a wide range of pathogenic fungi in vitro were tested for their efficacy against damping-off of wheat caused by Rhizoctonia solani Kühn in the greenhouse. These isolates were further studied for their modes of action against R. solani in vitro. The modes of action tested included antibiosis, production of siderophores, extracellular enzymes, production of hydrogen cyanide (HCN) and antibiotic resistance. The twenty most promising bacterial isolates identified from the in vitro screening reduced acetylene to ethylene, produced indole-3-acetic acid and siderophores, one isolate solubilized phosphate, and 9 inhibited the growth of R. solani. These twenty isolates enhanced growth of maize and wheat above the Uninoculated Control under greenhouse conditions. The growth enhancements varied with bacterial isolate x crop species interactions, which identified five different isolates for each of the two crops. Relative to the Uninoculated Control, the best five isolates significantly (P = 0.001) enhanced the growth of maize and wheat at all fertilizer levels for a number of growth parameters: increased chlorophyll levels and heights of maize, shoot dry biomass of maize and wheat, and enhanced root development of maize in the greenhouse. Inoculation of maize and wheat with the two most promising isolates identified from the field trial for each crop, in combination with 65% and 50% of the recommended amount of nitrogenous fertilizer for maize and wheat, respectively, caused the same increases in shoot biomass as the Fully Fertilized Control. Application of a combination of the best bacterial isolates and 35% nitrogenous fertilizer resulted in the same or greater shoot dry biomass and yields of both maize and wheat under field conditions. Shoot dry biomass of wheat increased by 75% above the Uninoculated Control and 30% above the Fully Fertilized Control. The wheat yield increased by 95% above the Uninoculated Control and 43% above the Fully Fertilized Control. Seed inoculation with the best isolates combined with 35% N increased yields of maize by 41% above the Uninoculated Control and 15% above the Fully Fertilized Control. The best isolates significantly (P < 0.001) increased plant height, chlorophyll levels and shoot biomass of maize relative to the Uninoculated Control. There was a positive correlation between chlorophyll level and yield, chlorophyll level and shoot dry biomass, height and shoot dry biomass and height and yield of maize at P = 0.01 with r values of 0.87, 0.77, 0.92 and 0.81, respectively. The isolates that exhibited multiple plant-growth promoting traits in vitro, increased shoot biomass of both maize and wheat in the greenhouse and field, and caused yield increases in the two crops under field conditions. Five B. subtilis isolates inhibited the growth of some of the pathogenic fungi tested in vitro up to 95%. Seed inoculation with the same isolates significantly (P = 0.001) suppressed R. solani damping-off of wheat under greenhouse conditions and exhibited multiple mechanisms of disease control in vitro. The use of microbial inoculants in combination with low doses of nitrogenous fertilizers can enhance crop production without compromising the yields. The B. subtilis isolates obtained in this study can effectively control R. solani damping-off of wheat, fix nitrogen and enhance plant growth. The use of microbial inoculants can contribute to the integrated production of cereal crops with reduced nitrogenous fertilizer inputs, as a key component of sustainable agriculture. Key words: Free-living bacteria; plant growth-promotion; diazotrophs; biological nitrogen fixation; phosphate-solubilization; siderophores; indole-3-acetic acid; biocontrol; damping-off / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2007. / Härtill 5 uppsatser.

Made available in DSpace on 2014-06-11T19:32:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-04-29Bitstream added on 2014-06-13T19:43:45Z : No. of bitstreams: 1 meneghin_sp_dr_rcla.pdf: 500401 bytes, checksum: 1dbb48df89774c8c8a05446c822888e1 (MD5) / Universidade Estadual Paulista (UNESP) / Nas usinas, no início da safra, a obtenção de matéria-prima de boa qualidade é maximizada com a aplicação de fitorreguladores, os quais aumentam o teor de sacarose da cana-de-açúcar. Em áreas onde eles são aplicados, tem se observado melhor desenvolvimento e perfilhamento das plantas. Avaliou-se aqui o efeito da aplicação dos fitorreguladores Ethrel e Moddus sobre o crescimento da cana-de-açúcar, de forma direta e indiretamente, através da modificação da microbiota rizosférica. Além disso, objetivou-se também avaliar o uso de rizobactérias, isoladas dos experimentos com fitorreguladores, para o biocontrole de doenças e seus possíveis mecanismos de ação. Os efeitos dos fitorreguladores sobre os microrganismos do solo foram avaliados em meios de cultura acrescidos de Ethrel e Moddus em concentrações de 0 a 1000 ppm. Estes fitorreguladores foram aplicados via foliar e via solo para análise do desenvolvimento da cana-de-açúcar (variedades RB72454, RB835486 e RB855156) em casa-de-vegetação, utilizando-se solo sem tratamento e tratado com brometo de metila. Após dez meses, foram avaliadas a brotação, altura e matéria seca da parte aérea e das raízes das plantas cultivadas. Rizobactérias foram isoladas dos solos contidos nos vasos e avaliadas in vitro quanto à capacidade de controle de fungos fitopatogênicos (Thielaviopsis paradoxa, Fusarium spp. e Hendersonina sacchari), e in vivo, quanto à capacidade de promoção de crescimento de plântulas de cana-de-açúcar. Alguns mecanismos de ação das rizobactérias foram também estudados, como produção de ácido indol acético, ácido cianídrico, sideróforos e solubilização de fosfato inorgânico. Constatou-se que as populações de fungos foram mais sensíveis à adição dos fitorreguladores do que outros grupos de microrganismos, com redução... / For sugar and alcohol industries, at the start of harvesting, to obtain good quality raw material is potentially possible with the application of plant regulators, which have a role in natural sugar cane maturation, increasing sucrose content. In areas where they have been applied, better plant development and shooting have been observed. The aim here was to evaluate the application of plant regulators Ethrel and Moddus on sugar cane growth, not only in a direct way, but also indirectly, through the modification of rhizosphere microorganisms. Besides, this work also aimed the evaluation of rhizobacteria isolated from the experiments using plant regulators upon the disease biocontrol and their action mechanisms in this respect. The effects of plant regulators upon the soil microorganisms were verified in culture media where Ethrel and Moddus were added in concentrations ranging from 0 to 1000 ppm, while the effects of these substances (applied in leaves and in soil) upon the sugar cane development (varieties RB72454, RB835486 and RB855156) were surveyed in greenhouse, using soil without treatment and treated with methyl bromide. After a ten-month period, the experiments were finished, and sprouting, height and aerial part and root dry matter were analyzed. Soil samples were taken from the pots for rhizobacteria isolation, which were evaluated initially in vitro regarding their ability to control plant pathogenic fungi (Thielaviopsis paradoxa, Fusarium spp. and Hendersonina sacchari), and in vivo, regarding their ability to promote sugar cane growth. Some action mechanisms were also studied, as indol acetic acid, cyanide acid and siderophore production and inorganic phosphate solubilization. It was verified that the fungi populations were more sensitive to the addition of plant regulators than other microorganisms, reducing their colony-forming unit (CFU)...(Complete abstract, click electronic access below)

Orientador: Samia Maria Tauk-Tornisielo / Banca: Sandra Regina Ceccato Antonini / Banca: Regina Teresa Rosim Monteiro / Banca: Antonio Ismael Bassinelo / Banca: Carlos Renato Corso / Resumo: Nas usinas, no início da safra, a obtenção de matéria-prima de boa qualidade é maximizada com a aplicação de fitorreguladores, os quais aumentam o teor de sacarose da cana-de-açúcar. Em áreas onde eles são aplicados, tem se observado melhor desenvolvimento e perfilhamento das plantas. Avaliou-se aqui o efeito da aplicação dos fitorreguladores Ethrel e Moddus sobre o crescimento da cana-de-açúcar, de forma direta e indiretamente, através da modificação da microbiota rizosférica. Além disso, objetivou-se também avaliar o uso de rizobactérias, isoladas dos experimentos com fitorreguladores, para o biocontrole de doenças e seus possíveis mecanismos de ação. Os efeitos dos fitorreguladores sobre os microrganismos do solo foram avaliados em meios de cultura acrescidos de Ethrel e Moddus em concentrações de 0 a 1000 ppm. Estes fitorreguladores foram aplicados via foliar e via solo para análise do desenvolvimento da cana-de-açúcar (variedades RB72454, RB835486 e RB855156) em casa-de-vegetação, utilizando-se solo sem tratamento e tratado com brometo de metila. Após dez meses, foram avaliadas a brotação, altura e matéria seca da parte aérea e das raízes das plantas cultivadas. Rizobactérias foram isoladas dos solos contidos nos vasos e avaliadas in vitro quanto à capacidade de controle de fungos fitopatogênicos (Thielaviopsis paradoxa, Fusarium spp. e Hendersonina sacchari), e in vivo, quanto à capacidade de promoção de crescimento de plântulas de cana-de-açúcar. Alguns mecanismos de ação das rizobactérias foram também estudados, como produção de ácido indol acético, ácido cianídrico, sideróforos e solubilização de fosfato inorgânico. Constatou-se que as populações de fungos foram mais sensíveis à adição dos fitorreguladores do que outros grupos de microrganismos, com redução...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: For sugar and alcohol industries, at the start of harvesting, to obtain good quality raw material is potentially possible with the application of plant regulators, which have a role in natural sugar cane maturation, increasing sucrose content. In areas where they have been applied, better plant development and shooting have been observed. The aim here was to evaluate the application of plant regulators Ethrel and Moddus on sugar cane growth, not only in a direct way, but also indirectly, through the modification of rhizosphere microorganisms. Besides, this work also aimed the evaluation of rhizobacteria isolated from the experiments using plant regulators upon the disease biocontrol and their action mechanisms in this respect. The effects of plant regulators upon the soil microorganisms were verified in culture media where Ethrel and Moddus were added in concentrations ranging from 0 to 1000 ppm, while the effects of these substances (applied in leaves and in soil) upon the sugar cane development (varieties RB72454, RB835486 and RB855156) were surveyed in greenhouse, using soil without treatment and treated with methyl bromide. After a ten-month period, the experiments were finished, and sprouting, height and aerial part and root dry matter were analyzed. Soil samples were taken from the pots for rhizobacteria isolation, which were evaluated initially in vitro regarding their ability to control plant pathogenic fungi (Thielaviopsis paradoxa, Fusarium spp. and Hendersonina sacchari), and in vivo, regarding their ability to promote sugar cane growth. Some action mechanisms were also studied, as indol acetic acid, cyanide acid and siderophore production and inorganic phosphate solubilization. It was verified that the fungi populations were more sensitive to the addition of plant regulators than other microorganisms, reducing their colony-forming unit (CFU)...(Complete abstract, click electronic access below) / Doutor

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.

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.

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.