Bacillus amyloliquenaciense FZB42 ist ein bekanntes Pflanzenwachstum-stimulierendes Rhizobakterium. Es produziert neben einer Vielzahl an Sekundärmetaboliten mit antibakterieller und antifungaler Wirkung, auch das Pflanzenhormon IAA. Obwohl viele dieser Mechanismen diskutiert werden, ist wenig darüber bekannt, auf welche Weise die Bakterien das Pflanzenwachstum fördern. In dieser Arbeit wurde eine Transposonmutagenese mithilfe des ‘mariner-transposons’ durchgeführt, und so eine Transposonbibliothek erstellt. Diese wurde dann auf geeignete Phänotypen untersucht, um die Gene zu finden, welche bestimmte Phänotypen verursachen. So konnten drei Mutanten erzeugt werden, die auf Grund der gestörten Biofilmbildung und der Fähigkeit zu schwärmen die Pflanzenwurzeln nicht mehr kolonialisieren konnten. Eine solche degU-Mutante, welche in der Biofilmbildung und ‚Swarming’ defizitär war und zwei Mutanten (yusV und pabB), die eine Beeinträchtigung in der Biofilmbildung aufwiesen, konnten durch Komplementation und Retransformation bestätigt werden. Mithilfe des Lemna-Biosystems und anderer Analysen mit A. thaliana konnten drei Gene bei B. amyloliqufaciens FZB42 gefunden werden, die wichtig für die Förderung des Pflanzenwachstums sind. Koloniesierungsexperimente der Wurzeln von A. thaliana mit diesen Mutanten zeigten deutlich verändertes Wachstum, verglichen mit dem Wildtypstamm. Ein weiteres Ziel dieser Arbeit war es neue Antibiotika in Mutanten, die in ihren nicht-ribosomalen Synthesen blockiert sind, zu finden. So konnten durch die Untersuchungen der Transposonbibliothek der Mutanten zwei neue Antibiotika entdeckt werden. Genauere Analysen dieser Antibiotika bestätigten, dass es sich um ein neues Bacteriocin (Amylocyclicin A) und ein neues Thiazol/Oxazole-modifiziertes Microcin (Plantazolicin) handelt. Die abschließenden Arbeiten beschäftigten sich dann mit Untersuchungen von Genen, welche für die Produktion von Substanzen gegen Nematoden verantwortlich sind. Hierbei konnten vier Mutanten gefunden werden, die durch eine Transposoninsertion eine schlechtere. / Bacillus amyloliqufaciens FZB42 has been known as PGPR which has an impressive effect to improve plant growth. It produces not only vast array of secondary metabolites with antibacterial and antifungal activities, but also produces the plant hormone IAA. Although many mechanisms have been elucidated, our knowledge about basic molecular mechanisms responsible for its beneficial action is far from complete. In this study, transposon mutagenesis based on mariner tranposon was applied to generate tranposon library which then was screened to identify the genes involved in plant growth-promoting activity. Three mutants that were impaired in their ability to colonize plant surface due to defects in biofilm formation and swarming motility were found. One mutant (degU mutant) showed defect in biofilm formation and swarming motility, as well, two mutants (yusV mutant and pabB mutant) impaired in biofilm formation were confirmed by complementation and retransformation. Screening by the Lemna biosystem and further assays with A. thaliana revealed three genes responsible for reduction in plant growth promoting activity of B. amyloliqufaciens FZB42. Colonization studies of these mutants in A. thaliana roots revealed patterns different to the wild type. A further issue pursued in this study was to discover new antibiotics using a mutant which has been blocked in its nonribosomally pathway. Screening of tranposon librabries from this mutant led to the finding of two novel ribosomally synthesized antibiotics. Further characterization revealed that these new antibiotics belonged to a novel bacteriocin (Amylocyclicin A) and a novel thiazole/oxazole-modified microcin (Plantazolicin). Last work in this study was looking for genes responsible for nematocidal production. Four mutants which showed reduction in nematocidal activity due to transposon insertion were found.

<p>  </p> <p><em>Sclerotinia sclerotiorum </em>is a soilborne pathogen of soybean that causes Sclerotinia stem rot, alternatively called white mold. Sclerotinia stem rot can cause significant yield losses under cool and wet environmental conditions. Two biofungicides, <em>Coniothyrium minitans </em>and <em>Bacillus amyloliquefaciens, </em>are currently available and labeled to limit or suppress <em>S. sclerotiorum</em> in soybean. These biofungicides can be applied in place of synthetic foliar fungicides to provide an alternative mode of action for the control of Sclerotinia stem rot. However, limited information is available regarding the efficacy of <em>C. minitans </em>and <em>B. amyloliquefaciens </em>as biocontrol agents of <em>S. sclerotiorum </em>in soybean and the sensitivity of the biofungicides biological activity on <em>S. sclerotiorum </em>to pesticides commonly used in soybean production systems. This research aims to provide management recommendations for <em>S. sclerotiorum </em>in soybean using <em>C. minitans </em>and <em>B. amyloliquefaciens </em>and to develop guidelines for how to incorporate the biofungicides into an established soybean pest management program. To assess the effectiveness of <em>C. minitans </em>and <em>B. amyloliquefaciens </em>as biocontrol agents of <em>S. sclerotiorum </em>dual culture, amended media, and soil plate assays were conducted along with experiments in the growth chamber and field. The presence of a distinct inhibition zone surrounding the <em>B. amyloliquefaciens </em>colony in the dual culture assay and the absence of mycelial growth on the media plates amended with <em>B. amyloliquefaciens </em>confirmed that the bacteria can control the mycelial growth of <em>S. sclerotiorum </em>through antibiosis. The absence of an inhibition zone surrounding the <em>C. minitans </em>isolate in the dual culture assay along with the degradation of sclerotia following treatment with <em>C. minitans </em>in the soil plate assay indicates an inability to limit the mycelial growth of <em>S. sclerotiorum </em>and confirms that the primary mode of action is mycoparasitism. In the growth chamber, <em>B. amyloliquefaciens</em> at 14.03 L/ha applied using the dip method significantly reduced Sclerotinia stem rot lesion length when compared to the non-treated control and resulted in the lowest lesion area under the disease progress curve (lAUDPC). When <em>B. amyloliquefaciens </em>and <em>C. minitans </em>were applied in the field, no differences were observed between treatments for soybean moisture, test weight, or yield. To evaluate the sensitivity of <em>B. amyloliquefaciens </em>and <em>C. minitans</em> biological activity on <em>S. sclerotiorum </em>to pesticides commonly used in soybean production systems a poison plate assay as well as soil plate, growth chamber, and field experiments were conducted. In the poison plate assay <em>C. minitans </em>was most sensitive to the preemergence herbicide flumioxazin and the synthetic fungicides boscalid and fluazinam, while <em>B. amyloliquefaciens </em>was sensitive only to the synthetic fungicide fluazinam. In the soil plate assay the mycoparasitic activity of <em>C. minitans </em>on sclerotia of <em>S. sclerotiorum </em>was sensitive to flumioxazin, metribuzin, glyphosate, picoxystrobin, and boscalid. In the controlled environment experiments, none of the pesticides tested decreased the efficacy of <em>B. amyloliquefaciens</em>. There were no significant interactions between <em>C. minitans </em>and <em>B. amyloliquefaciens </em>with preemergence herbicides, postemergence herbicides, and synthetic fungicides for soybean moisture, test weight, and yield. This research demonstrates that <em>B. amyloliquefaciens </em>and <em>C. minitans </em>are effective biocontrol agents of <em>S. sclerotiorum </em>in soybean. However, antagonistic relationships exist between the biofungicides and certain preemergence, postemergence, and synthetic fungicides used in soybean production systems.</p>