Determinants of crop diversity and composition in Enset-coffee agroforestry homegardens of Southern Ethiopia

Households in much of the tropics depend for their livelihoods on the variety and continued production of food and other products that are provided by their own farms. In such systems, maintenance of agrobiodiversity and ensuring food security are important for the well being of the population. The enset-coffee agroforestry homegardens of Southern Ethiopia that are dominated by two native perennial crops, Coffee (Coffea arabica L.) and Enset (Enset ventricosum Welw. Cheesman), are examples of such agricultural systems. This study was conducted in Sidama administrative zone of Southern Ethiopia to determine the factors that influence the diversity and composition of crops in the systems. Data were collected from 144 sample homegardens selected from four districts. Stepwise multiple regression analysis was used to relate indices of crop diversity and area share of major crops with the physical and socioeconomic factors. The study revealed that socioeconomic factors, mainly proximity to markets, affected negatively crop species richness. The production area of the main crops enset and coffee decreased with increasing proximity to market and road while that of maize and khat increased. At household level, farm size had a significant effect on area share of enset and coffee. As farm size increased the share of the cash crop, coffee increased but that of the staple, enset declined. Enset, which is the backbone of the system in terms of food security, is declining on small farms and the share of monoculture maize system is increasing. The trend towards declining agrobiodiversity, and reduction in the production area of the main perennial crops and their gradual replacement with monoculture fields could make the systems liable to instability and collapse. As these sites are high potential agricultural areas, intensification can be achieved by integrating high-value and more productive crops, such as fruits, spices and vegetables, while maintaining the integrated and complex nature of the systems.

Heterochromatin und epigenetische Kontrolle der Centromere in Dictyostelium discoideum

Heterochromatin Protein 1 (HP1) is an evolutionarily conserved protein required for formation of a higher-order chromatin structures and epigenetic gene silencing. The objective of the present work was to functionally characterise HP1-like proteins in Dictyostelium discoideum, and to investigate their function in heterochromatin formation and transcriptional gene silencing. The Dictyostelium genome encodes three HP1-like proteins (hcpA, hcpB, hcpC), from which only two, hcpA and hcpB, but not hcpC were found to be expressed during vegetative growth and under developmental conditions. Therefore, hcpC, albeit no obvious pseudogene, was excluded from this study. Both HcpA and HcpB show the characteristic conserved domain structure of HP1 proteins, consisting of an N-terminal chromo domain and a C-terminal chromo shadow domain, which are separated by a hinge. Both proteins show all biochemical activities characteristic for HP1 proteins, such as homo- and heterodimerisation in vitro and in vivo, and DNA binding activtity. HcpA furthermore seems to bind to K9-methylated histone H3 in vitro. The proteins thus appear to be structurally and functionally conserved in Dictyostelium. The proteins display largely identical subnuclear distribution in several minor foci and concentration in one major cluster at the nuclear periphery. The localisation of this cluster adjacent to the nucleus-associated centrosome and its mitotic behaviour strongly suggest that it represents centromeric heterochromatin. Furthermore, it is characterised by histone H3 lysine-9 dimethylation (H3K9me2), which is another hallmark of Dictyostelium heterochromatin. Therefore, one important aspect of the work was to characterise the so-far largely unknown structural organisation of centromeric heterochromatin. The Dictyostelium homologue of inner centromere protein INCENP (DdINCENP), co-localized with both HcpA and H3K9me2 during metaphase, providing further evidence that H3K9me2 and HcpA/B localisation represent centromeric heterochromatin. Chromatin immunoprecipitation (ChIP) showed that two types of high-copy number retrotransposons (DIRS-1 and skipper), which form large irregular arrays at the chromosome ends, which are thought to contain the Dictyostelium centromeres, are characterised by H3K9me2. Neither overexpression of full-length HcpA or HcpB, nor deletion of single Hcp isoforms resulted in changes in retrotransposon transcript levels. However, overexpression of a C-terminally truncated HcpA protein, assumed to display a dominant negative effect, lead to an increase in skipper retrotransposon transcript levels. Furthermore, overexpression of this protein lead to severe growth defects in axenic suspension culture and reduced cell viability. In order to elucidate the proteins functions in centromeric heterochromatin formation, gene knock-outs for both hcpA and hcpB were generated. Both genes could be successfully targeted and disrupted by homologous recombination. Surprisingly, the degree of functional redundancy of the two isoforms was, although not unexpected, very high. Both single knock-out mutants did not show any obvious phenotypes under standard laboratory conditions and only deletion of hcpA resulted in subtle growth phenotypes when grown at low temperature. All attempts to generate a double null mutant failed. However, both endogenous genes could be disrupted in cells in which a rescue construct that ectopically expressed one of the isoforms either with N-terminal 6xHis- or GFP-tag had been introduced. The data imply that the presence of at least one Hcp isoform is essential in Dictyostelium. The lethality of the hcpA/hcpB double mutant thus greatly hampered functional analysis of the two genes. However, the experiment provided genetic evidence that the GFP-HcpA fusion protein, because of its ability to compensate the loss of the endogenous HcpA protein, was a functional protein. The proteins displayed quantitative differences in dimerisation behaviour, which are conferred by the slightly different hinge and chromo shadow domains at the C-termini. Dimerisation preferences in increasing order were HcpA-HcpA << HcpA-HcpB << HcpB-HcpB. Overexpression of GFP-HcpA or a chimeric protein containing the HcpA C-terminus (GFP-HcpBNAC), but not overexpression of GFP-HcpB or GFP-HcpANBC, lead to increased frequencies of anaphase bridges in late mitotic cells, which are thought to be caused by telomere-telomere fusions. Chromatin targeting of the two proteins is achieved by at least two distinct mechanisms. The N-terminal chromo domain and hinge of the proteins are required for targeting to centromeric heterochromatin, while the C-terminal portion encoding the CSD is required for targeting to several other chromatin regions at the nuclear periphery that are characterised by H3K9me2. Targeting to centromeric heterochromatin likely involves direct binding to DNA. The Dictyostelium genome encodes for all subunits of the origin recognition complex (ORC), which is a possible upstream component of HP1 targeting to chromatin. Overexpression of GFP-tagged OrcB, the Dictyostelium Orc2 homologue, showed a distinct nuclear localisation that partially overlapped with the HcpA distribution. Furthermore, GFP-OrcB localized to the centrosome during the entire cell cycle, indicating an involvement in centrosome function. DnmA is the sole DNA methyltransferase in Dictyostelium required for all DNA(cytosine-)methylation. To test for its in vivo activity, two different cell lines were established that ectopically expressed DnmA-myc or DnmA-GFP. It was assumed that overexpression of these proteins might cause an increase in the 5-methyl-cytosine(5-mC)-levels in the genomic DNA due to genomic hypermethylation. Although DnmA-GFP showed preferential localisation in the nucleus, no changes in the 5-mC-levels in the genomic DNA could be detected by capillary electrophoresis.

Framework for middleware executed on mobile devices

Die ubiquitäre Datenverarbeitung ist ein attraktives Forschungsgebiet des vergangenen und aktuellen Jahrzehnts. Es handelt von unaufdringlicher Unterstützung von Menschen in ihren alltäglichen Aufgaben durch Rechner. Diese Unterstützung wird durch die Allgegenwärtigkeit von Rechnern ermöglicht die sich spontan zu verteilten Kommunikationsnetzwerken zusammen finden, um Informationen auszutauschen und zu verarbeiten. Umgebende Intelligenz ist eine Anwendung der ubiquitären Datenverarbeitung und eine strategische Forschungsrichtung der Information Society Technology der Europäischen Union. Das Ziel der umbebenden Intelligenz ist komfortableres und sichereres Leben. Verteilte Kommunikationsnetzwerke für die ubiquitäre Datenverarbeitung charakterisieren sich durch Heterogenität der verwendeten Rechner. Diese reichen von Kleinstrechnern, eingebettet in Gegenstände des täglichen Gebrauchs, bis hin zu leistungsfähigen Großrechnern. Die Rechner verbinden sich spontan über kabellose Netzwerktechnologien wie wireless local area networks (WLAN), Bluetooth, oder UMTS. Die Heterogenität verkompliziert die Entwicklung und den Aufbau von verteilten Kommunikationsnetzwerken. Middleware ist eine Software Technologie um Komplexität durch Abstraktion zu einer homogenen Schicht zu reduzieren. Middleware bietet eine einheitliche Sicht auf die durch sie abstrahierten Ressourcen, Funktionalitäten, und Rechner. Verteilte Kommunikationsnetzwerke für die ubiquitäre Datenverarbeitung sind durch die spontane Verbindung von Rechnern gekennzeichnet. Klassische Middleware geht davon aus, dass Rechner dauerhaft miteinander in Kommunikationsbeziehungen stehen. Das Konzept der dienstorienterten Architektur ermöglicht die Entwicklung von Middleware die auch spontane Verbindungen zwischen Rechnern erlaubt. Die Funktionalität von Middleware ist dabei durch Dienste realisiert, die unabhängige Software-Einheiten darstellen. Das Wireless World Research Forum beschreibt Dienste die zukünftige Middleware beinhalten sollte. Diese Dienste werden von einer Ausführungsumgebung beherbergt. Jedoch gibt es noch keine Definitionen wie sich eine solche Ausführungsumgebung ausprägen und welchen Funktionsumfang sie haben muss. Diese Arbeit trägt zu Aspekten der Middleware-Entwicklung für verteilte Kommunikationsnetzwerke in der ubiquitären Datenverarbeitung bei. Der Schwerpunkt liegt auf Middleware und Grundlagentechnologien. Die Beiträge liegen als Konzepte und Ideen für die Entwicklung von Middleware vor. Sie decken die Bereiche Dienstfindung, Dienstaktualisierung, sowie Verträge zwischen Diensten ab. Sie sind in einem Rahmenwerk bereit gestellt, welches auf die Entwicklung von Middleware optimiert ist. Dieses Rahmenwerk, Framework for Applications in Mobile Environments (FAME²) genannt, beinhaltet Richtlinien, eine Definition einer Ausführungsumgebung, sowie Unterstützung für verschiedene Zugriffskontrollmechanismen um Middleware vor unerlaubter Benutzung zu schützen. Das Leistungsspektrum der Ausführungsumgebung von FAME² umfasst: • minimale Ressourcenbenutzung, um auch auf Rechnern mit wenigen Ressourcen, wie z.B. Mobiltelefone und Kleinstrechnern, nutzbar zu sein • Unterstützung für die Anpassung von Middleware durch Änderung der enthaltenen Dienste während die Middleware ausgeführt wird • eine offene Schnittstelle um praktisch jede existierende Lösung für das Finden von Diensten zu verwenden • und eine Möglichkeit der Aktualisierung von Diensten zu deren Laufzeit um damit Fehlerbereinigende, optimierende, und anpassende Wartungsarbeiten an Diensten durchführen zu können Eine begleitende Arbeit ist das Extensible Constraint Framework (ECF), welches Design by Contract (DbC) im Rahmen von FAME² nutzbar macht. DbC ist eine Technologie um Verträge zwischen Diensten zu formulieren und damit die Qualität von Software zu erhöhen. ECF erlaubt das aushandeln sowie die Optimierung von solchen Verträgen.