Universiẗat, Diss., 2001--Bremen.

This thesis was performed within the research project “TrophinOak”, which addresses the impact of multitrophic interactions on the pedunculate oak (Quercus robur) clone DF159. In this frame, the present work focuses on the genetic and physiological mechanisms ruling the interaction of the mycorrhiza helper bacterium (MHB) Streptomyces sp. AcH 505 with microcuttings of DF159 either alone or in presence of the ectomycorrhizal fungus Piloderma croceum or the fungal leaf pathogen oak powdery mildew Microsphaera alphitoides. The work consists of 3 chapters. Chapter 1 characterises the growth of AcH 505 and P. croceum in a soil-based culture system used within the TrophinOak project. Besides the establishment and evaluation of quantification methods of these microorganisms by quantitative real-time PCR, the impact of the soil microbial community and the oak on the bacterium-fungus interaction was investigated, and AcH 505 and P. croceum were visualized by scanning electron microscopy. It was observed that the presence of the soil microorganisms and the oak both affect the bacterium-fungus interaction, and that P. croceum enhances the growth of AcH 505. Chapter 2 presents a study with the oak, AcH 505 and the EM fungus P. croceum, enabling to disentangle the direct effect of the MHB on the oak from the indirect one via the EM symbiosis. The used approach was transcriptomic based on RNA sequencing. It was shown that i) differential gene expression occurred between root and the distant leaf tissues (local vs. systemic effects), different developmental stages and treatments, suggesting that oak specifically coordinates its gene expression patterns, and ii) that genes related to plant growth, defence and DNA modification were dominant among the differential expressed genes, suggesting that these processes play essential roles in both symbiotic interactions investigated. Chapter 3 represents a second transcriptome study, addressing how AcH 505 suppresses powdery mildew infection in oak by analysing RNA Sequencing data from singly- and coinoculated oaks. This study combined the systemic impact of the root associated bacterium with local effects of the leaf pathogen, thereby linking belowground and aboveground interactions. Systemic defence response is induced by the bacterium and further enhanced upon pathogen challenge, suggesting that on the leaf level, some bacterial effectors are recognized as harmful for the plant.

For the first time the transcriptional reprogramming of distinct root cortex cells during the arbuscular mycorrhizal (AM) symbiosis was investigated by combining Laser Capture Mirodissection and Affymetrix GeneChip® Medicago genome array hybridization. The establishment of cryosections facilitated the isolation of high quality RNA in sufficient amounts from three different cortical cell types. The transcript profiles of arbuscule-containing cells (arb cells), non-arbuscule-containing cells (nac cells) of Rhizophagus irregularis inoculated Medicago truncatula roots and cortex cells of non-inoculated roots (cor) were successfully explored. The data gave new insights in the symbiosis-related cellular reorganization processes and indicated that already nac cells seem to be prepared for the upcoming fungal colonization. The mycorrhizal- and phosphate-dependent transcription of a GRAS TF family member (MtGras8) was detected in arb cells and mycorrhizal roots. MtGRAS shares a high sequence similarity to a GRAS TF suggested to be involved in the fungal colonization processes (MtRAM1). The function of MtGras8 was unraveled upon RNA interference- (RNAi-) mediated gene silencing. An AM symbiosis-dependent expression of a RNAi construct (MtPt4pro::gras8-RNAi) revealed a successful gene silencing of MtGras8 leading to a reduced arbuscule abundance and a higher proportion of deformed arbuscules in root with reduced transcript levels. Accordingly, MtGras8 might control the arbuscule development and life-time. The targeting of MtGras8 by the phosphate-dependent regulated miRNA5204* was discovered previously (Devers et al., 2011). Since miRNA5204* is known to be affected by phosphate, the posttranscriptional regulation might represent a link between phosphate signaling and arbuscule development. In this work, the posttranscriptional regulation was confirmed by mis-expression of miRNA5204* in M. truncatula roots. The miRNA-mediated gene silencing affects the MtGras8 transcript abundance only in the first two weeks of the AM symbiosis and the mis-expression lines seem to mimic the phenotype of MtGras8-RNAi lines. Additionally, MtGRAS8 seems to form heterodimers with NSP2 and RAM1, which are known to be key regulators of the fungal colonization process (Hirsch et al., 2009; Gobbato et al., 2012). These data indicate that MtGras8 and miRNA5204* are linked to the sym pathway and regulate the arbuscule development in phosphate-dependent manner. / Die Leguminose Medicago truncatula (gehört zur Gattung des Schneckenklees) ist in der Lage sowohl eine Symbiose mit stickstofffixierenden Bakterien (Rhizobien), als auch mit Mykorrhiza-Pilzen einzugehen. Der Mykorrhiza-Pilz Rhizophagus irregularis dringt in die Wurzelrindenzellen ein und bildet Strukturen aus, die als Arbuskeln bezeichnet werden. Diese ermöglichen den Transfer von Nährstoffen, wie Phosphat in die Wurzelzellen. Die Pflanze liefert hingegen bis zu 20 % ihrer Photosyntheseprodukte an den Pilz. Da die Lebenszeit der Arbuskeln nur wenige Tage beträgt, können Wurzelrindenzellen mehrere Arbuskeln nacheinander beherbergen. Somit können neben arbuskelhaltigen, auch nicht-arbuskelhaltige Zellen in kolonisierten Wurzeln auftreten. Die nicht-arbuskelhaltigen Zellen beeinträchtigen die Sensitivität von Genregulationsanalysen, wenn die Genregulation während der Mykorrhiza-Symbiose anhand von ganzen kolonisierten Wurzeln untersucht wird. In dieser Arbeit konnte eine Zelltyp-spezifische Analyse der Genregulation von arbuskelhaltigen und nicht-arbuskelhaltigen Zellen durchgeführt, und eine Erhöhung der Sensitivität erreicht werden. Mittels Laser Capture Microdissection wurden Zellen aus Gefrierschnitten von Wurzeln isoliert. Aus den so gewonnen Zellen konnte RNA von ausreichender Qualität und Quantität extrahiert werden, um das Transkriptom der beiden Zelltypen mittels Mikroarrayhybridisierung zu untersuchen. Transkriptionsfaktoren (TFs) spielen wahrscheinlich eine Schlüsselrolle in der Umprogrammierung von Wurzelzellen während der Mykorrhiza-Symbiose. Daher wurde die Genregulation von TF-Genen in den zwei Zelltypen gezielt untersucht. Anhand von quantitativer RT-PCR und Promoter-Reporter-Fusionen wurde die differentielle Expression von mehreren TF-Transkripten in den verschiedenen Zelltypen bestätigt. Die Charakterisierung eines potentiellen GRAS TF (MtGRAS8) konnte eine stark Symbiose- und Phosphat-abhängige Induktion von Transkripten bestätigt werden. Mutanten mit verringerter MtGras8 Transkriptmenge wiesen eine verringerte Arbuskelzahl und deformierte Arbuskeln auf. MtGras8 scheint daher an der Arbuskelentwicklung beteiligt zu sein. Vorherige Experimente zeigten, dass MtGras8 Transkripte, von der Phosphat-regulierten MikroRNA5204* geschnitten werden (Devers et al., 2011). Dies konnte durch Überexpression der MikroRNA5204* in vivo bestätigt werden. Weiterhin konnten Protein-Protein-Interaktionen von MtGras8 mit bekannten GRAS TFs (NSP1, NSP2, RAM1) nachgewiesen und daraus eine Verbindung zu bekannten Symbiose-induzierten Signalkaskaden geschlossen werden. In dieser Arbeit wurde erstmals die Umprogrammierung von nicht-arbuskelhaltigen Zellen untersucht und neue Regulationselemente für die Kontrolle der Arbuskelentwicklung, wie MtGRAS8 und MikroRNA5204*, charakterisiert.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet,2008. / Härtill 4 uppsatser.

Techn. University, Diss., 2004--Darmstadt.

The aim of this thesis was to detect whether achlophyllous orchid Corallorhiza trifida associates different fungal taxa in different habitats. Our analyses were conducted using molecular and fylogenetic methods based on DNA sequencing.

No description available.

Alder (Alnus glutinosa L. Gaertn.) is a species of a pioneer plant usually colonizing sites in the early stage of ecological succession, such as spoil heaps after open-cast brown coal mining in the Sokolov mining district in north western Bohemia, Czech Republic. These spoil heaps are very poor in nutrients available for plants, yet alders grow in a mutualistic relationship with actinomycetes Frankia, which live in root nodules of the alder plants. Frankia are able of fixing atmospheric nitrogen (N2) to ammonia (NH3) molecules, which can be then assimilate by alders, via enzyme nitrogenase. Thus, in the early stages of succession, alders have a competitive advantage to other non- fixing plant species living only on nitrates (NO3 - ) and ammonia ions (NH4 + ) present in soils. The aim of this study was to conduct and assess two greenhouse experiments. The first experiment studied the response of alder growth to presence of Frankia and arbuscular mycorrhizal fungi of order Glomerales. The performance of alder growth was significantly higher when alders were inoculated with both Frankia and mycorrhizal fungi in comparison to when alders have grown on their own or only with a mycorrhizal symbiont - both on 14 and 60 years old soils from Sokolov mines. In the second experiment, soil pH and iron (Fe) and...

This thesis was performed within the research project “TrophinOak”, which addresses the impact of multitrophic interactions on the pedunculate oak (Quercus robur) clone DF159. In this frame, the present work focuses on the genetic and physiological mechanisms ruling the interaction of the mycorrhiza helper bacterium (MHB) Streptomyces sp. AcH 505 with microcuttings of DF159 either alone or in presence of the ectomycorrhizal fungus Piloderma croceum or the fungal leaf pathogen oak powdery mildew Microsphaera alphitoides. The work consists of 3 chapters. Chapter 1 characterises the growth of AcH 505 and P. croceum in a soil-based culture system used within the TrophinOak project. Besides the establishment and evaluation of quantification methods of these microorganisms by quantitative real-time PCR, the impact of the soil microbial community and the oak on the bacterium-fungus interaction was investigated, and AcH 505 and P. croceum were visualized by scanning electron microscopy. It was observed that the presence of the soil microorganisms and the oak both affect the bacterium-fungus interaction, and that P. croceum enhances the growth of AcH 505. Chapter 2 presents a study with the oak, AcH 505 and the EM fungus P. croceum, enabling to disentangle the direct effect of the MHB on the oak from the indirect one via the EM symbiosis. The used approach was transcriptomic based on RNA sequencing. It was shown that i) differential gene expression occurred between root and the distant leaf tissues (local vs. systemic effects), different developmental stages and treatments, suggesting that oak specifically coordinates its gene expression patterns, and ii) that genes related to plant growth, defence and DNA modification were dominant among the differential expressed genes, suggesting that these processes play essential roles in both symbiotic interactions investigated. Chapter 3 represents a second transcriptome study, addressing how AcH 505 suppresses powdery mildew infection in oak by analysing RNA Sequencing data from singly- and coinoculated oaks. This study combined the systemic impact of the root associated bacterium with local effects of the leaf pathogen, thereby linking belowground and aboveground interactions. Systemic defence response is induced by the bacterium and further enhanced upon pathogen challenge, suggesting that on the leaf level, some bacterial effectors are recognized as harmful for the plant.

No description available.