Successful chickpea production in western Canada typically requires multiple applications of fungicides to minimize the severity of Ascochyta blight (AB) caused by <i>Ascochyta rabiei</i>. Although planting resistant cultivars could be economical and environmentally safer than fungicide usage, varieties with a high level of resistance are not available. The objectives of this research were i) to determine the effect of different seeding arrangement treatments on ascochyta blight severity and seed yield of two cultivars (moderately resistant and susceptible) of kabuli chickpea; ii) to compare one and four fungicide applications at recommended and reduced rates and their impact on disease severity and cost; and iii) to assess organ-specific reaction to AB in chickpea in leaves, stems and pods of 12 desi and 12 kabuli varieties that are of economic significance to western Canada. <p>Treatments significantly influenced AB severity on both moderately resistant and susceptible cultivars in a season with a severe epidemic. Seed yield was significantly influenced by treatments for both varieties in both years. Contrast analyses revealed that four fungicide applications significantly reduced the AB severity for both varieties in a season with a severe epidemic and for the susceptible variety in a season with a moderate epidemic. Seed yield of both varieties was significantly higher under four fungicide applications compared to a single application. Solid seeding and paired row arrangements did not differ in their effect on seed yield and AB severity for both varieties in both years, except that the susceptible variety benefited from paired row planting with respect to seed yield and reduced AB severity in the season with a severe epidemic. Reducing fungicide rates and seeding rate could reduce the cost of cultivation without significantly affecting disease control and yield. Economic assessment revealed that in a severe epidemic season, the gross returns were high for the moderately resistant variety under four fungicide applications than one fungicide application. Gross returns for the susceptible variety were higher under four fungicide applications in both years.<p>There were differences among varieties for AB severity on leaves, stems and pods, seed yield and 1000 seed weight at all site-years tested. The variation was greater in kabuli varieties than desi varieties. AB severity on leaves, stems and pods was lower under high fungicide regimes, with few exceptions. Varieties with a fern leaf type had lower AB severity than those with unifoliate leaves. There was a positive correlation among AB severity on leaves, stems and pods. No differences in organ-specific reaction were observed.

Successful chickpea production in western Canada typically requires multiple applications of fungicides to minimize the severity of Ascochyta blight (AB) caused by <i>Ascochyta rabiei</i>. Although planting resistant cultivars could be economical and environmentally safer than fungicide usage, varieties with a high level of resistance are not available. The objectives of this research were i) to determine the effect of different seeding arrangement treatments on ascochyta blight severity and seed yield of two cultivars (moderately resistant and susceptible) of kabuli chickpea; ii) to compare one and four fungicide applications at recommended and reduced rates and their impact on disease severity and cost; and iii) to assess organ-specific reaction to AB in chickpea in leaves, stems and pods of 12 desi and 12 kabuli varieties that are of economic significance to western Canada. <p>Treatments significantly influenced AB severity on both moderately resistant and susceptible cultivars in a season with a severe epidemic. Seed yield was significantly influenced by treatments for both varieties in both years. Contrast analyses revealed that four fungicide applications significantly reduced the AB severity for both varieties in a season with a severe epidemic and for the susceptible variety in a season with a moderate epidemic. Seed yield of both varieties was significantly higher under four fungicide applications compared to a single application. Solid seeding and paired row arrangements did not differ in their effect on seed yield and AB severity for both varieties in both years, except that the susceptible variety benefited from paired row planting with respect to seed yield and reduced AB severity in the season with a severe epidemic. Reducing fungicide rates and seeding rate could reduce the cost of cultivation without significantly affecting disease control and yield. Economic assessment revealed that in a severe epidemic season, the gross returns were high for the moderately resistant variety under four fungicide applications than one fungicide application. Gross returns for the susceptible variety were higher under four fungicide applications in both years.<p>There were differences among varieties for AB severity on leaves, stems and pods, seed yield and 1000 seed weight at all site-years tested. The variation was greater in kabuli varieties than desi varieties. AB severity on leaves, stems and pods was lower under high fungicide regimes, with few exceptions. Varieties with a fern leaf type had lower AB severity than those with unifoliate leaves. There was a positive correlation among AB severity on leaves, stems and pods. No differences in organ-specific reaction were observed.

Vagal afferents form the primary gut-to-brain neural axis and are thought to communicate negative feedback signals to the central nervous system to attenuate consummatory behaviors by promoting satiation and possibly satiety. The expansive and fluid nature of the gastrointestinal organs has made it methodologically challenging to decipher the negative feedback signals, and how the signals are disseminated or converged within the central feeding systems. We sought to understand the anatomical relationship and organization between the terminal fields of the peripheral axonal projections and the central axonal projections of gastrointestinal (GI) vagal afferents for clues about what and how information is communicated along the gut-brain axis. Here, we quantified the density and distribution of peripheral and central GI vagal axonal projections in neurotrophin-4 deficient (KO) and control mice. KO mice exhibited a 75 and 55% reduction in small intestinal vagal mucosal afferents, proximally distally, and no significant reduction of mucosal vagal afferents in the stomach, compared to controls. Previous characterization, similarly, reported a >70% reduction in small intestinal vagal muscle afferents and no loss of muscle afferents in the stomach. Centrally, KO mice exhibited an increase in central terminal axonal projections in the medial nucleus tractus solitarius. Our findings support previous hypotheses that neurotrophin-4 exerts an organ-specific regulation of development of gastrointestinal vagal afferents innervation. Furthermore, our findings highlight the dynamic relationship between the peripheral and central axonal projections of vagal afferents.

No description available.

Verticillium is a soil-borne fungus that is one of the world's foremost plant pathogens. Commercial plant grafting suggests that resistance occurs in the root; this conflicts with decades of research indicating that resistance occurs in the stem. The goal of this thesis work was to use an alternative approach to determine the location of resistance by expressing the Ve1 gene using organ-specific promoters. Promoter sequences for the stem-specific gene, Ribulose 1,5-bisphosphate carboxylase oxygenase small chain 2A (Rbsc2A), and root-specific gene, Tobacco Mosaic Virus Induced (TMVi) were taken from the Sol Genomics Network (SGN) database, cloned into constructs with the Ve1 gene and susceptible tomato germplasm was transformed using Agrobacterium tumefaciens. Preliminary results suggest that resistance may not be localized and expression of the Ve1 gene in either the root or the stem is sufficient to develop whole plant resistance to the Verticillium pathogen.

Bei der Aufrechterhaltung des homeostatischen Gleichgewichts und der peripheren Selbst-Toleranz spielen CD4+CD25+ regulatorische T-Zellen (Tregs) eine wichtige Rolle. In Vorarbeiten wurden Subpopulationen von murinen CD4+ Tregs identifiziert, die sich durch die Expression des Integrins alphaE auszeichnen. Diese alphaE+ Treg Subpopulationen weisen einen Effektor/Memory-ähnlichen Phänotyp auf. In der vorliegenden Dissertation wurde untersucht, welche Bedingungen zur Entwicklung von alphaE+ Effektor/Memory Tregs in vivo führen und aus welchen Vorläuferzellen sie entstehen. Dabei zeigte sich, dass es sich bei den alphaE+ Tregs um Effektor/Memory T-Zellen handelt, die unter physiologischen Bedingungen in vivo ein hohes Maß an Zellteilung aufweisen, welche zum Teil abhängig von der bakteriellen Besiedelung des Darms ist. Darüber hinaus wurde beobachtet, dass alphaE+ Tregs nach oraler, antigen-spezifischer Aktivierung in den darm-assozierten lymphoiden Geweben sowohl aus konventionellen naiven CD4+ T-Zellen, als auch aus thymus-generierten naiven CD4+CD25+ Tregs entstehen können. Zusammenfassend deuten die erzielten Ergebnisse darauf hin, dass das spezifische mukosale Mikroenvironment sowohl die Expansion als auch Konvertierung von Tregs fördert und so eine wichtige Rolle für die Aufrechterhaltung der Homeostase von alphaE+Foxp3+ Tregs spielt. Zudem wurde in dieser Arbeit die Ausbildung von gewebespezifischen Homingrezeptor-Phänotypen von naiven CD4+CD25+ Tregs untersucht. In in vitro Kultur-Systemen zeigte sich, dass selektive Modulation von Tregs, ähnlich wie bei konventionellen T-Zellen, die Induktion von organspezifischen Migrationseigenschaften ermöglicht. So konnte eine effiziente Wanderung von Tregs in den Darm ausgelöst werden. Diese Daten legen den Schluss nahe, dass die Herstellung von Tregs mit spezifischen Wanderungseigenschaften eine Option für therapeutische Anwendungen in der adoptiven T-Zell Therapie sein könnte. / Regulatory CD4+CD25+ T cells (Tregs) play an important role in immune homeostasis and in the maintenance of self-tolerance. Previously, subsets of murine CD4+ Tregs characterised by expression of the integrin alphaE had been identified. These alphaE+ Treg subsets display an effector/memory-like phenotype. In the present study the circumstances favouring in vivo generation of effector/memory-like alphaE+ Tregs were analysed. The results presented here show that alphaE+ effector/memory-like Treg subsets contain a large fraction of cycling cells under physiologic conditions in healthy mice. This in vivo proliferation depended, at least in part, on intestinal commensal microflora. Furthermore, it was observed that alphaE+ Tregs not only developed by differentiation of naive-like CD4+CD25+ Tregs, but were also generated de novo from naive CD4+ T cells in the gut-associated lymphoid tissue upon oral antigen delivery. Taken together, these results indicate that the mucosal microenvironment favours both expansion and conversion of Tregs and thereby represents an important mechanism for the homeostatic maintenance of alphaE+Foxp3+ Tregs. In addition, susceptibility of naive CD4+CD25+ Tregs to acquire tissue-specific homing receptor phenotypes was investigated. In vitro culture systems demonstrated that Tregs, similarly to conventional T cells, could be configured with organ-selective homing properties allowing efficient targeting into the gut. These results suggest that generation of Tregs with specific homing properties for therapeutic purposes in adoptive T cell therapy might be a feasible option.

Immunologische Toleranz ist eine elementare Eigenschaft des Immunsystems, die primär durch die klonale Deletion autoreaktiver T-Zellen im Thymus gewährleistet wird. Neben diesem als zentrale Toleranz bezeichneten Mechanismus, verfügt ein Organismus gleichzeitig über periphere Toleranzmechanismen wie Ignoranz, Anergie und regulatorische T-Zellen. Trotz dieser Kontrollmechanismen können in bestimmten Situationen autoreaktive CD4+ und CD8+ T-Zellen aktiviert werden und meistens zu örtlich und zeitlich begrenzten Autoimmunreaktionen führen. Ursache hierfür kann die hormonelle Regulation oder das gewebespezifische Vorkommen eines Selbsttantigens sein. Am Beispiel von HSP60-kreuzreaktiven CD8+ T-Zellen konnte gezeigt werden, dass der Transfer dieser T-Zellen in Tiere zu einer Entzündung des Dünndarms aber nicht des Dickdarms führt, obwohl das Selbstantigen im letzteren wesentlich stärker exprimiert wird. Die Gewebespezifität der Autoimmunpathologie konnte durch die in den Organen unterschiedliche, proteasomale Antigenprozessierung, erklärt werden. Proteinbiochemische und immunologische Analysen ergaben, dass sich die 20S Proteasomen verschiedener Organe strukturell und funktionell deutlich unterscheiden und somit jedes Gewebe ein individuelles Repertoire von MHC-Klasse I restringierten Peptiden präsentiert. Damit wurde ein weiterer Mechanismus entdeckt, durch den Reaktivität von protektiven und pathologischen CD8+ T-Zellen kontrolliert wird. / Immunological tolerance which is primarily mediated by the clonal deletion of autoreactive T cells in the thymus is a key feature of the immune system. Besides this central tolerance, several mechanisms act also in the periphery including ignorance, anergy and regulatory T cells. Despite all these checkpoints, autoreactive CD4+ and CD8+ T cells may still be activated causing local and time restricted autoimmune-reactions. This may refer primarily to self-antigens which are hormonally regulated or tissue-specifically expressed. Adoptive transfer of crossreactive, hsp60-specific CD8+ T cells into mice induced an local inflammation of the small intestine but not the colon despite elevated expression of hsp60 in the latter organ. The pathology could be explained by the finding that the proteasomal antigen processing varies between different organs. Biochemical and immunological analyses revealed that 20S proteasomes of different organs vary in their structural and functional properties indicating that every tissue displays an individual and distinct repertoire of MHC class I peptides. This represents a new mechanism by which the activity of protective and pathological CD8+ T cell responses may be controlled.

Nitrogen (N), an essential plant macronutrient, is among the most limiting factors of crop yield. To sustain modern agriculture, N is often amended in soil in the form of chemical N fertilizer, a major anthropogenic contributor to nutrient pollution that affects climate, biodiversity and human health. To achieve agricultural sustainability, a comprehensive understanding of the regulation of N response in plants is required, in order to engineer crops with higher N use efficiency. Recently, epigenetic mechanisms, such as histone modifications, have gained increasing importance as a new layer of regulation of biological processes. However, our understanding of how epigenetic processes regulate N uptake and assimilation is still in its infancy. To fill this knowledge gap, we first performed a meta-analysis that combined functional genomics and network inference approaches to identify a set of N-responsive epigenetic regulators and predict their effects in regulating epigenome and transcriptome during plant N response. Our analysis suggested that histone modifications could serve as a regulatory mechanism underlying the global transcriptomic reprogramming during plant N response. To test this hypothesis, I applied chromatin immunoprecipitation-sequencing (ChIP-Seq) to monitor the genome-wide changes of four histone marks (H3K27ac, H3K4me3, H3K36me3 and H3K27me3) in response to N supply in tomato plants, followed by RNA-Seq to profile the transcriptomic changes. To investigate the organ specificity of histone modifications, I assayed shoots and roots separately. My results suggest that up to two-thirds of differentially expressed genes (DEGs) are modified in at least one of the four histone marks, supporting an integral role of histone modification in regulating N response. I observed a synergistic modification of active histone marks (H3K27ac, H3K4me3 and H3K36me3) at gene loci functionally relevant to N uptake and assimilation. Surprisingly, I uncovered a non-canonical role of H3K27me3, which is conventionally associated with repressed genes, in modulating active gene expression. Interestingly, such regulatory role of H3K27me3 is specifically associated with highly expressed genes or low expressed genes, depending on the organ context. Overall, I revealed the multi-faceted role of histone marks in mediating the plant N response, which will guide breeding and engineering of better crops with higher N use efficiency