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Made available in DSpace on 2015-03-26T13:51:46Z (GMT). No. of bitstreams: 1 01 - capa_abstract.pdf: 113068 bytes, checksum: ac8d58045328d0ccbe13706fe9c3dede (MD5) Previous issue date: 2008-09-12 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The heavy oil (10 20ºAPI) is more than half of world oil reserves, there are predictions that in 2025 are the main source of oil in the world. The difficulties in exploration this kind of oil are associated with unfavorable characteristics such as density and viscosity (100 to 10.000 cP). With the need to improve the utilization of reservoir - in view of the relatively low levels of efficiency of traditional techniques - it has expanded the research to develop new techniques for recovery, especially Microbialy Enhanced Oil Recovery (MEOR). In the recovery of oil contained in sand-pack columns, two dimensionless variables are reported as important: the capillary number and ratio of viscosity. As the increases capillary number, the residual oil decreases. This can be achieved by reducing the interfacial tension between the fluid of residual oil. Several authors have reported the tensions interfaciais ultra between biosurfactant and hydrocarbons, including heavy oil. In the present work, tested the hypothesis that the biosurfactant produced on condition of reservoir, by two isolates of Bacillus subtilis from the collection of culture of the Laboratório de Biotecnologia e Biodiversidade para o Meio Ambiente (LBBMA / DMB / UFV), are capable to remove heavy oil in sand-pack columns. Initially, the isolates of Bacillus subtilis LBBMA 155 and Bacillus subtilis subsp. spizizenii LBBMA 258 were evaluated as to the growth and biosurfactant production in response to temperature and salinity. A central composite rotatable design (CCRD) was used, with temperature and salinity being independent variable. The results were analyzed using Response Surface Methodology (RSM) and showed that the those isolates were able of growth and biosurfactant production of under anaerobic conditions. The growth was influenced by temperature and biosurfactant production was influenced by temperature and salinity. After that, the biosurfactant produced by those isolates, alone or mixed, were analyzed as the ability to remove heavy oil trapped in sand-pack columms. The injection of three volumes of porous extract biossurfactante (EB) produced by Bacillus subtilis LBBMA 155 and Bacillus subtilis subsp. spizizenii LBBMA 258 removed 13.55% and 17.42%, respectively, from residual oil (14 ºAPI). The mixed of extracts from biosurfactant produced by those two isolates showed intermediate values of DMC and recovery of residual oil reported for each. Summarize, the extracts from biosurfactant (EB) produced under anaerobic conditions by Bacillus subtilis LBBMA 155 e Bacillus subtilis subsp. spizizenii LBBM 258 are effective in recovery heavy oil nuclei of the sand-pack columms and have potencial for use Microbialy Enhanced Oil Recovery. / Os óleos pesados (10-20 ºAPI) constituem mais da metade das reservas mundiais de petróleo, havendo previsões de que em 2025 sejam a principal fonte de petróleo no mundo. As dificuldades na exploração desse tipo de óleo estão associadas a características desfavoráveis, como densidade e viscosidade (100 a 10000 cP). Com a necessidade de melhorar o aproveitamento dos reservatórios tendo em vista os níveis relativamente baixos de eficiência das técnicas tradicionais tem-se ampliado as pesquisas para o desenvolvimento de novas técnicas de recuperação, destacando-se a recuperação avançada de petróleo melhorada por microrganismos (Microbially enhanced oil recovery MEOR). Na recuperação de óleos contidos em meios porosos, duas variáveis adimensionais são relatadas como importantes: o número capilar e a razão de viscosidade. À medida que o número capilar aumenta, o óleo residual decresce. Isso pode ser obtido por meio da redução da tensão interfacial entre o fluido de arraste e o óleo residual. Diversos autores têm reportado a obtenção de tensões interfaciais ultrabaixas entre biossurfactantes e hidrocarbonetos, incluindo o óleo pesado. Neste trabalho, testou-se a hipótese de que os biossurfactantes produzidos, em condição de reservatório, por dois isolados de Bacillus subtilis pertencentes à Coleção de Culturas do Laboratório de Biotecnologia e Biodiversidade para o Meio Ambiente (LBBMA/DMB/UFV), são capazes de mobilizar óleo pesado em sistemas porosos de areia. Inicialmente, os isolados de Bacillus subtilis LBBMA 155 e BacilIus subtilis spizizenii LBBMA 258 foram avaliados quanto ao crescimento e produção de biossurfactante em resposta a variações de temperatura e salinidade. Para tanto, foi empregada a Metodologia de Superfície de Resposta (MSR) através do delineamento experimental Composto Central Rotacional (DCCR), e observou-se que esses isolados foram capazes de produzir biossurfactantes em anaerobiose, sendo o crescimento influenciado pela temperatura e a produção de biossurfactante influenciada pela temperatura e pela salinidade. Posteriormente, os biossurfactantes produzidos por esses isolados, sozinhos ou em misturas, foram avaliados quanto à capacidade de mobilização de óleo pesado retido em núcleos porosos de areia. A injeção de três volumes porosos do extrato de biossurfactante (EB) produzidos por Bacillus subtilis LBBMA 155 e Bacillus subtilis subsp. spizizenii LBBMA 258 removeram 13,55% e 17,42%, respectivamente, de petróleo residual (14 ºAPI). A mistura dos extratos de biossurfactantes produzidos por esses dois isolados mostrou valores intermediários de DMC e recuperação de petróleo residual reportados para cada um. Sinteticamente, os extratos de biossurfactantes (EB) produzidos em anaerobiose por B. subtilis LBBMA 155 e B. subtilis subsp. spizizenii LBBMA 258 são efetivos na mobilização de óleo pesado em núcleos porosos de areia e possuem potencial de utilização na recuperação avançada de petróleo melhorada por microrganismos.

Estudos envolvendo o emprego de novas técnicas de esterilização têm apresentado nítido crescimento nos últimos anos como alternativa aos processos convencionais. A grande possibilidade consiste no emprego do plasma como agente de esterilização. Essa tecnologia tem considerável potencial para desenvolver o meio mais eficiente e seguro de esterilização de artigos termossensíveis com ênfase para a indústria farmacêutica e médica e até mesmo em outras áreas industriais. O objetivo deste trabalho foi investigar a influência de alguns parâmetros de processos por plasma e correlacionar sua eficácia com processos por óxido de etileno. Neste trabalho, estudos foram realizados empregando tecnologia de esterilização por plasma usando reator Reactive Ion Etching (RIE). Os valores aplicados de potências de rádio-frequência, a 13,56 MHz foram 25 W, 50 W, 100 W e 150 W. Os gases testados foram oxigênio puro e misturas de oxigênio e peróxido de hidrogênio (190/10, 180/20 e 160/40 sccm) e fluxo constante de 200 sccm, pressão de 0,100 torr e temperatura abaixo de 60°C. Esterilizador por óxido de etileno foi empregado a 450 mg/L (Oxyfume 2002®), 55°C, 60% de umidade e -0,65 e 0,60 kgf/cm2 de pressão. Os indicadores biológicos empregados foram constituídos de esporos de Bacillus subtilis var. niger ATCC 9372, inoculados em lamínulas de vidro de 18 x 18 mm e discos de papel de 13 mm de diâmetro em uma carga de 2,0 x 107 UFC/suporte. Os tempos de exposição aos processos por plasma foram de 3 a 120 minutos. Reduções progressivas da contagem inicial de microrganismos foram observadas nos valores D: 215,91,55,55,9,19 e 2,91 minutos para processos por plasma com oxigênio puro, a 25 W, 50 W, 100 W e 150 W, respectivamente. Misturas de oxigênio e peróxido de hidrogênio apresentaram os seguintes valores D: 190/10 sccm (6,41 min), 180/20 sccm (6,47 min) e 160/40 sccm (4,02 min), a 100 W e 190/10 sccm (1,47 min), 180/20 sccm (3,11 min) e 160/40 sccm (1,94 min), a 150 W. Processos empregando óxido de etileno apresentaram valor D de 2,86 minutos. Análises por Microscopia Eletrônica de Varredura demonstraram danos causados ao córtex dos esporos. Sistema empregando plasma como principal agente de esterilização apresentou-se efetivo em desafios com indicadores biológicos. Os processos por plasma provaram ser a mais apropriada tecnologia de esterilização de materias termossensíveis e com grande potencial para substituir os métodos convencionais de esterilização em futuro próximo. / Studies involving new sterilization techniques have increased in the past few years as alternatives to conventional processes. The great possibility consists in the use of plasma as sterilization agent. This technology has the enormous potential to develop a more efficient and safer means of sterilization at thermo-sensitive matters, focusing the pharmaceutical and medical industry - even though it can be applied to other industrial areas. The aim of the present study was to investigate the influence at some parameters of plasma processes and correlate the effectiveness plasma with ethylene oxide sterilizer. In this work, studies were performed taking into account a plasma sterilization technology using a Reactive Ion Etching (RIE) reactor. Power was applied at 13.56 MHz using a 6 inch diameter electrode. The gases tested were pure oxygen and oxygen-hydrogen peroxide mixtures (190/10, 180/20, and 160/40 sccm), and gas flow held constant 200 sccm, pressure at 0.100 torr and radio-frequency power at 25 W, 50 W, 100 W, and 150 W and temperature below 60°C. Ethylene oxide sterilizer were performed using 450 mg/L (Oxyfume 2002®) at 55°C, 60% humidity and -0.65 and 0.60 kgf/cm2 pressure. The biological indicator used was Bacillus subtilis var. niger ATCC 9372, witch was inoculated in glass carries (18 x 18 mm) and paper discs (13 mm diameter) in a load of 2.0 x 107 CFU/support. The exposition times were 3 to 120 minutes. Progressive reductions of the initial microbial count could be observed in the D values witch were 215.91, 55.55, 9.19, and 2.91 minutes for pure oxygen plasma at 25 W, 50 W, 100 W and 150 W, respectively. Oxygen-hydrogen peroxide mixtures plasma showed D values: 190/10 sccm (6.41 min), 180/20 sccm (6.47 min) and 160/40 sccm (4.02 min) at 100 W and 190/10 sccm (1.47 min), 180/20 sccm (3.11 min) and 160/40 sccm (1.94 min) at 150 W. Ethylene oxide processes showed D value to 2.86 minutes. Scanning Electron Microscopy analyses showed some damage on the spore cortex. Processes using plasma as main sterilization agent are presented effective in challenge with biological indicators. The plasma proved to be the most appropriate sterilization technology in thermosensitive matters and to have a great potential to replace conventional sterilization methods in the near future.

Le lin, principale plante à fibres cultivée en France possède un intérêt industriel pour la qualité de ses fibres. Les cultures de lin sont régulièrement attaquées par un champignon tellurique, Fusarium oxysporum f. sp. lini (Fol), responsable des plus grandes pertes dans les cultures de lin. Les PGPR (Plant Growth Promoting Rhizobacteria) sont des bactéries réputées pour leurs capacités à améliorer la croissance et le développement des plantes, mais également pour leur pouvoir compétiteur au sein de la rhizosphère et leur aptitude à induire une réponse immunitaire chez les plantes. Parallèlement, l’utilisation de SDP (Stimulateur des Défenses des Plantes), molécules capables d’éliciter les mécanismes de défenses des plantes est une autre alternative pour limiter l’utilisation des pesticides. Dans le cadre de ce projet mené sur deux variétés de lin, Aramis et Mélina, il a pu être montré que Fol induisait un remodelage de la paroi au niveau des racines et des tiges, impliquant les hémicelluloses et les pectines, seulement deux jours après inoculation avec le champignon. L’utilisation de la souche ATCC 6633 de Bacillus subtilis comme agent de biocontrôle a permis de réduire de façon significative l’apparition des symptômes de la fusariose. Il a également été montré qu’en plus d’avoir un effet fongicide sur Fol, cette bactérie est capable d’induire l’expression de deux gènes de défense (Pathogenesis-Related) codant pour une β-(1,3)-glucanase (PR-2) et codant pour une chitinase-like (CTL-10), de gènes impliqués dans la voie des phénylpropanoïdes (PHENYLALANINE AMONIA LYASES, PAL-3 et PAL-4) et dans le remodelage pariétal (PECTIN METHYLESTERASE-3, PME-3) au niveau racinaire. Des analyses biochimiques ont également permis de montrer que B. subtilis provoque des modifications se traduisant par un renforcement pariétal au niveau des tiges chez les deux variétés. Enfin, l’association de la PGPR avec une molécule élicitrice (pregnénolone sulfate) a eu un effet synergique sur l’expression de gènes de défense. / In France, flax (Linum usitatissumum) is a principal fibers crop. Fusarium oxysporum f sp lini (Fol), a soil-borne fungus, is responsible for the major losses in crop yield. PGPR (Plant Growth Promoting Rhizobacteria) are known for their abilities to promote plant growth and health. These bacteria are also good competitors in the rhizosphere and can induce a plant defense response. The use of compounds able to elicit plant defense mechanisms is also an alternative to limit the use of pesticides. In this project, it has been shown that F. oxysporum f. sp. lini induces only two days after inoculation cell wall remodeling in the root and the stem involving hemicelluloses and pectins on two flax varieties, Aramis and Mélina, . The use of the Bacillus subtilis strain ATCC 6633 as biocontrol agent significantly reduced fusarium wilt appearance. In addition to its antifungal effect against Fol, this bacteria is able to induce the expression of two Pathogenesis-Related genes coding for a β-(1,3)-glucanase (PR-2) and a chitinase-like (CTL-10), genes involved in the phenylpropanoid pathway (PHENYLALANINE AMONIA LYASES, PAL-3 and PAL-4) and also in cell wall remodeling (PECTIN METHYLESTERASE-3, PME-3) in the root. Biochemical analyses show that B. subtilis causes modifications resulting in cell wall reinforcement in the stem in both varieties. Finally, the association of B. subtilis with an elicitor (pregnenolone sulfate) had a synergistic effect on the expression of defense-related genes.

Maintaining cell envelope integrity is of vital importance for all microorganisms. Not surprisingly, evolution has shaped conserved protein protection networks that connect stress perception, transmembrane signal transduction, and mediation of cellular responses upon cell envelope stress. The phage shock protein (Psp) stress response is one such conserved protection network. Most knowledge about the Psp response derives from studies in the Gram-negative model bacterium Escherichia coli, where the Psp system consists of several well-defined protein components. Homologous systems were identified in representatives of the Proteobacteria, Actinobacteria, and Firmicutes. However, the Psp system distribution in the microbial world remains largely unknown. By carrying out a large-scale, unbiased comparative genomics analysis, we found components of the Psp system in many bacterial and archaeal phyla and describe that the predicted Psp systems deviate dramatically from the known prototypes. The core proteins PspA and PspC have been integrated into various (often phylum-specifically) conserved protein networks during evolution. Based on protein domain-based and gene neighborhood analyses of pspA and pspC homologs, we built a natural classification system for Psp networks in bacteria and archaea. We validate our approach by performing a comprehensive in vivo protein interaction study of Psp domains identified in the Gram-positive model organism Bacillus subtilis and found a strong interconnected protein network. Our study highlights the diversity of Psp domain organizations and potentially diverse functions across the plethora of the microbial landscape, thus laying the ground for studies beyond known Psp functions in underrepresented organisms.

Over the course of the last decades, the continuous exposure of bacteria to antibiotics—at least in parts due to misprescription, misuse, and misdosing—has led to the widespread development of antimicrobial resistances. This development poses a threat to the available medication in losing their effectiveness in treating bacterial infections. On the drug development side, only minor advances have been made to bring forward novel therapeutics. In addition to increasing the efforts and approaches of tapping the natural sources of new antibiotics, synthetic approaches to developing novel antimicrobials are being pursued. In this study, BDTL049 was rationally designed using knowledge based on the properties of natural antibiotics. BDTL049 is a carbosilane dendritic system with bow-tie type topology, which has antimicrobial activity at concentrations comparable to clinically established natural antibiotics. In this report, we describe its mechanism of action on the Gram-positive model organism Bacillus subtilis. Exposure to BDTL049 resulted in a complex transcriptional response, which pointed toward disturbance of the cell envelope homeostasis accompanied by disruption of other central cellular processes of bacterial metabolism as the primary targets of BDTL049 treatment. By applying a combination of whole-cell biosensors, molecular staining, and voltage sensitive dyes, we demonstrate that the mode of action of BDTL049 comprises membrane depolarization concomitant with pore formation. As a result, this new molecule kills Gram-positive bacteria within minutes. Since BDTL049 attacks bacterial cells at different targets simultaneously, this might decrease the chances for the development of bacterial resistances, thereby making it a promising candidate for a future antimicrobial agent.

Das Ziel der Arbeit war die Entwicklung einer Begrünungsmethode für Gleisbette mit schneller Vegetationsentwicklung. Zur Begrünung wurden Sedumpflanzen verwendet, die durch den Einsatz von Bacillus subtilis, Lactobacillus und Nährsubstrat (allein und kombiniert) in verschiedenen Vegetationssystemen auf dem nährstoffarmen Standort Gleisbett etabliert werden sollten. Die Aktivität der inokulierten Mikroorganismen wurde indirekt über den Einfluss auf die Vegetationsleistung ermittelt. Der Einsatz von B. subtilis und Lactobacillus bewirkte eine signifikante Wachstumsförderung der oberirdischen Pflanzenteile. Die Anwendung von Nährsubstrat als Bodenhilfsmittel war ebenfalls für das Pflanzenwachstum besonders wirksam, sowohl allein appliziert als auch in Kombination mit den Bakterien. Darüber hinaus führte die Applikation der Bakterien und des Nährsubstrates zu einem reduzierten Trockenstress auf Geotextilmatten. Die Wahl des Substrates war entscheidend für die Wirksamkeit der Bakterieninokulation. Jedoch hatten höhere Versuchstemperaturen und pH-Werte ebenfalls eine positive Wirkung auf das Pflanzenwachstum. Diese Einflüsse waren in Kombination mit Ziegelbruchsubstrat wesentlich stärker als mit den anderen getesteten Substrattypen. Das Ziegelbruchsubstrat kombiniert mit einer Bakterien- und Nährsubstratbehandlung bewährte sich als günstiges, umweltschonendes Begrünungsverfahren von Gleisbettanlagen. / The aim of the present study was to develop and improve existing methods for the remediation of rail tracks using soil borne bacteria. Through the use of Bacillus subtilis and Lactobacillus ssp. alone and in combination with a nutrient solution three different growth substrates were tested. The substrates were brick chips, textile mats, and mineral wool mats. Brick chips were tested along railway tracks in Munich but all three substrates were tested along an artificial rail track on the experimental station at the Humboldt University-Berlin. Plants selected for remediation belong to the genus Sedum, which is relatively tolerant to dry conditions. The use of Bacillus subtilis and Lactobacillus in combination with a nutrient solution improved plant growth significantly. Plants inoculated with bacteria showed increased growth during the first three months but after four months there was no longer any significant difference between treatments. The addition of nutrient solution alone improved plant growth. Plant growth was significantly different on all three substrates, whereas brick chips were the best substrate. The results of this study indicate that the quality of the substrate is the most important factor for remediation and greening of rail tracks. Both bacteria tested had only a limited effect. High temperature and pH resulted in larger Sedum plants in the open field. Brick chips are a cheap substrate which can be used for rail track greening. The rapid growth of plants can be influenced by the application of an additional nutrient solution as well as inoculation with Lactobacillus and/or B. subtilis.

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.

Biotische und abiotische Stressfaktoren mindern die Quantität und die Qualität landwirtschaftlicher Erzeugnisse. Sogar die kontrollierten Wachstumsbedingungen eines Gewächshauses tragen nur bedingt zur Minderung der schädlichen Auswirkungen von suboptimalen Wachstumsfaktoren bei. Die negativen Effekte, die durch die Entstehung suboptimaler Wachstumsbedingungen auftreten können, manifestieren sich oft erst nach geraumer Zeit. Aus diesem Grund ist es wichtig, Maßnahmen zu treffen, die diesen negativen Auswirkungen entgegenwirken und eine nachhaltige Produktion von Obst und Gemüse sichern. Um die Widerstandsfähigkeit der Pflanzen gegenüber den zahlreichen Stressfaktoren zu erhöhen, werden sogenannte Pflanzenstärkungsmittel verwendet. Die zahlreichen gärtnerischen Substrate, die bei der Kultivierung von Obst und Gemüse heutzutage unentbehrlich geworden sind, besitzen verschiedene chemische und physikalische Eigenschaften. Diese Eigenschaften unterliegen dem ständigen Einfluss von verschiedenen Faktoren, insbesondere dem Pflanzenwachstum. Anderseits beeinflussen gärtnerische Substrate die Entwicklung der Pflanzen. Die Fähigkeit eines Substrates zur Wasserhaltung, sowie zur Gewährleistung einer optimalen Versorgung mit pflanzlichen Nährstoffen während der kritischen Wachstumsperioden der Pflanzen kann die Produktivität der gärtnerischen Kulturen stark beeinflussen. Dynamische Veränderungen der Substrateigenschaften während des Pflanzenwachstums und Änderungen in physiologischen Reaktionen der gärtnerischen Pflanzen können als System betrachtet werden, und als System können sie, zwecks Optimierung der pflanzlichen Systeme, beeinflusst werden. Diese Einflussnahme kann durch Verwendung von Pflanzenstärkungsmitteln verschiedener Herkunft erreicht werden. In der Fachliteratur wird häufig über Pflanzenstärkungsmittel diskutiert. Dabei wird sehr oft außer Acht gelassen, dass jedes Pflanzenstärkungsmittel ein eigenes Wirkungsspektrum besitzt. Die Breite des Wirkungsspektrums eines Pflanzenstärkungsmittels ist begrenzt, weshalb die positiven Effekte des verwendeten Mittels häufig geringer ausfallen können als erwartet. Die Arbeitshypothese dieser Arbeit belegt, dass durch die Kombination verschiedener bioaktiver Stoffe mit Pflanzenstärkungsmitteln dieses Wirkungsspektrum erweitert werden kann. Aus diesen Zusammenhängen leitet sich das Interesse an Kombinationen verschiedener Pflanzenstärkungsmittel sowie anderer bioaktiver Stoffe ab. Diese Arbeit untersucht die biologischen Effekte verschiedener Kombinationen von bioaktiven Stoffen und Pflanzenstärkungsmitteln zur Stabilisierung pflanzlicher Systeme. Als bioaktive Stoffe werden in dieser Arbeit Laktate und Humate bezeichnet, wobei Mikroorganismen als Pflanzenstärkungsmittel eingesetzt wurden. Die Ergebnisse dieser Arbeit zeigen, dass ein kombinierter Einsatz von Mikroorganismen, Humaten und Laktaten zum Einen zum Erhalt der chemischen und physikalischen Eigenschaften gärtnerischer Substrate, zum Anderen zur Stabilisierung der Pflanzen gegenüber suboptimalen Wachstumsfaktoren beiträgt. Die Anwendung der Kombination von allen drei Komponenten auf Versuchsvarianten mit Gewächshausgurken als Modellpflanze zeigte, dass die Pflanzenproduktivität bei diesen Varianten am höchsten war. Die physiologischen Reaktionen der Gurkenpflanzen im Versuch mit modellierten Stressfaktoren wurden durch Chlorophyllfluoreszenz ermittelt. Die Ergebnisse zeigten, dass die Pflanzen, die mit allen drei Komponenten behandelt wurden, gegenüber suboptimalen Wachstumsbedingungen resistent sind. Die Resultate dieser Arbeit wurden in einem gärtnerischen Unternehmen approbiert. / Biotic and abiotic stress factors reduce the quality and quantity of the horticultural produce. Even controlled growing conditions of a greenhouse can not reduce all the negative influences of the suboptimal growing factors. The negative effects caused by the suboptimal growing factors manifest themselves only after substantial period of time. That is why it is important to take steps that counter these negative effects and enable sustainable production of fruits and vegetables. To improve the resistance of the plants against different stress factors, the plant strengtheners are applied. The horticultural substrates, which are used for cultivation of the fruits and vegetables, have become unalienable and possess the range of different physical and chemical properties. These properties are subjected to constant influence of different biotic and abiotic factors; especially those resulted from the plant growth itself. On the other hand, the substrates influence the growth of the horticultural plants. The water holding capacity of the substrate and its capacity to provide plants with nutrient elements during the critical vegetation periods can influence the productivity of the plant system. Dynamic changes of the substrate properties during the plant growth and changes in physiological reactions of horticultural plants can be viewed as a system, and as the system it can be influenced for the purposes of its optimization. This influence can be achieved by application of different plant strengtheners of different origin. The role of the plant strengthening agents is being discussed in the literature. The missing point in the discussion however is the fact that the plant strengtheners have their own activity spectrum. The activity spectrum of the bioactive substance is limited and expected positive effects of its application can be lesser that expected. The hypothesis of the thesis is based on the premise that combination of the different bioactive substances and plant strengthening agents can increase the activity spectrum. These interrelations cause an interest on combinations of different plant strengtheners and others bioactive substances. This thesis investigates biological effects different combinations of bioactive substances and plant strengtheners with an aim to stabilize plant systems. In this work, lactates and humates are described as bioactive compounds, whereas microorganisms are perceived as plant strengtheners. The results of this work show that the combined application of microorganisms, humates and lactates sustain chemical and physical properties of the horticultural substrates and at the same time stabilize plant systems under the suboptimal growing factors. The experiment variants with greenhouse cucumbers as a model plant treated with all three components showed the highest productivity. Physiological reactions of the cucumber plants were investigated through measurement of the chlorophyll fluorescence of the cucumber leaves in the experiments with modeled suboptimal growing factors. The results showed that the plants treated with a combination of humates lactates and microorganism were more resistant to suboptimal growing conditions than the plants on variants without treatment. The results of this work were approbated at a horticultural enterprise.

Untersuchungen wurden durchgeführt, um die Wirkung von B. subtilis und deren Metaboliten auf den Meloidogyne- und Pratylenchus-Befall und ihre Vermehrung festzustellen sowie um die möglichen Wirkmechanismen zu studieren. Substratbehandlungen mit B. subtilis FZB 24® führten zu einem höherenMeloidogyne-Befall und einer verstärkten Nematodenvermehrung. Trotz verstärktem Befall wurde das Pflanzenwachstum verbessert (induzierte Toleranz). Weiterhin wurden durch "antibiotikafreie" Kulturfiltrate (KF) aus der bakterielen Übergangs- und stationären Phase ähnliche Wirkungen erzielt. Außerdem wurde eine systemische Wirkung von B. subtilis auf den Meloidogyne-Befall an Tomate nachgewiesen. B. subtilis bzw.die KF-behandelte Testpflanzen zeigten stärkere Anlockwirkung auf Meloidogyne-Larven (M. incognita, M. arenaria) als unbehandelte. KF (50, 10, 1%) von B. subtilis zeigten keine nematizide Wirkung auf die Meloidogyne-Larven. KNO3 als Trägersubstanz für das Bakterienpräparat besaß ähnliche Wirkungen auf den Meloidogyne-Befall und die Nematodenvermehrung.Ebenfalls wurde das Pflanzenwachstum durch KNO3-Zufuhr gefördert. Der Einsatz des nematodenfangenden Pilzes Arthrobotrys superba reduzierte den Meloidogyne-Befall (30% Gallenreduktion). Hingegen wurde durch die kombinierte Anwendung von A.superba und B. subtilis FZB 24® der Bekämpfungserfolg von A. superba reduziert. Die exogene Applikation von Phytohormonen bzw. Präkursoren zeigte keine Wirkung auf das Wachstum der Testpflanzen. Die Vermehrung von Meloidogyne wurde durch IAA und die Kombination von IAA und Kinetin gefördert. In den getesteten Konzentrationen der Phytohormone wurde keine Wirkung auf die Mortalität der Wurzelgallenälchen- (Meloidogyne-) Larven beobachtet. Der Gehalt von Enzymen (Chitinase, Glucanase und Peroxidase) aus dem Sproß behandelter Tomatenpflanzen wurde bestimmt. B. subtilis-Isolate (FZB 24® , S18) reduzierten die Population von Wurzelläsionsnematoden, Pratylenchus penetrans (nicht signifikant) [9 bzw. 15-20% pro Wurzelsystem bzw. g Wurzel]. Das Pflanzenwachstum wurde an befallenen Pflanzen durch beide Isolate verbessert (induzierte Resistenz). Es wurden keine Unterschiede zwischen den Bakterien-Isolaten festgestellt. KNO3 führte ebenfalls zu einer Verminderung der Nematodenpopulation. Die Ergebnisse werden hinsichtlich möglicher Wirkmechanismen des RhizobakteriumsB. subtilis und des nematodenfangendes PilzesA. superba zur Regulierung der Nematodenpopulation bei Tomate diskutiert. / Investigations were made to know about the effects of Bacillus subtilis and its metabolites on the infestation of tomato plants with root-knot and lesion nematodes. Further more, experiments were carried out to clear up the mode of actions ofB. subtilis and its culture filtrates on infestation of tomato seedlings and reproduction of root-knot nematodes. Substrate applications ofB. subtilis FZB 24® leaded to an increasement of infestations intensity and reproduction of root-knot nematodes (M. arenaria). Eventhough, bacterized and inoculated plants with root-knot nematodes showed better growth than with bacteria untreated plants (induced tolerance). In addition "antibiotica free" culture filtrates (CF) from transitional and stationary bacterial growth phase also promoted reproduction of root-knot nematodes. These CF elicitized tolerance of tomato plants towardsMeloidogyne too. It was proved that B. subtilis could induce a systemic reaction of tomato plants towards root-knot infestation. Besides that test plants treated with B. subtilis (cells) or CF were more attractive to Meloidogyne-Larvae (M.arenaria & M.incognita) than untreated once. CF in 50, 10 and 1% concentrations did not have a nematicidal effect on the root-knot larvae. KNO3 -the carrier of the bacterial preparate (B. subtilis FZB 24®) - had also the same effects on infestation and reproduction of root-knot nematodes. Plant growth was promoted due to application of KNO3. The use of nematode trapping fungus, Arthrobotrys superba gave some range of nematode (root-knot) control (30% gall reduction). Whereas, with the combination ofA. superba and B. subtilis FZB 24® the effect of the fungus was reduced. The application of exogenic phytohormones and precursores showed no effect on plant growth. Reproduction of Meloidogyne was promoted by IAA and combination of IAA and kinetin (not significant). In the tested concentrations of these phytohormones there was no direct mortality effect on root-knot larvae. Content of some enzymes (chitinase, glucanase and peroxidase) from shoot of treated tomato plants was determind. B. subtilis-Isolates (FZB 24® and S18) reduced the population of lesion nematodes,Pratylenchus penetrans in attacked plants (not significant) [9% per root system and 15-20% per g root]. The treatment improved the predisposition of the plants to lesion nematodes (induced resistance). Plant growth was also improved. There was no difference between the bacterial isolates in their effect. KNO3 reduced also nematode population. In general the results would be explained and discussed towards possible mode of actions of rhizobacterium B.subtilis and nematode trapping fungus A. superba.