Involvements of the Plant 3'-5' Exonuclease ERL1 in Chloroplast Ribosomal RNA Biogenesis and RNA Silencing Pathways

ERI-1 und ihm homologe Proteine sind 3‘-5‘ Exoribonukleasen mit konservierten Funktionen in der Regulation von RNA Silencing sowie der Prozessierung ribosomaler RNA. Caenorhabditis elegans ERI-1 (Enhanced RNAi 1) enthält eine konservierte ERI-1_3’hExo_like EXOIII-Domäne, die siRNAs in vitro bindet und degradiert, und deren Inaktivierung eine RNAi-Hypersensitivität zur Folge hat. ERI-1 ist phylogenetisch konserviert, und homologe Proteine wurden Reiche-übergreifend in einer Vielzahl von Modellorganismen identifiziert. RNA-Silencing-reprimierende Eigenschaften dieser Proteine wurden in einigen Fällen charakterisiert. Zusätzlich wurde für eine Untergruppe ERI-1-homologer Proteine eine Funktion in der Biogenese der 5.8S ribosomalen RNA aufgezeigt: Katalyse des letzten Prozessierungsschritts während der Reifung des 5.8S rRNA 3‘-Endes. Diese Doppelfunktion ERI-1-homologer Proteine schlägt eine interessante Brücke zwischen evolutionär weit entfernten auf nicht-codierender RNA basierenden Mechanismen. In dieser Arbeit werden Ergebnisse präsentiert, die Charakteristika des pflanzlichen ERI-1-Homologs ERL1 in verschiedenen regulatorischen Zusammenhängen zum Gegenstand haben. ERL1 lokalisiert in Chloroplasten und zeigt keinerlei messbare Aktivität in Bezug auf die Regulierung von RNA Silencing. Im Gegensatz dazu konnte gezeigt werden, dass ERL1 eine wichtige Rolle während der Reifung der chloroplastischen 5S rRNA spielt. ERL1-supprimierende bzw. -überexprimierende transgene Pflanzen, zeigen unterschiedliche phänotypische Aberrationen. Diese beinhalten vielfarbige Blätter, reduziertes Wachstum und Fruchtbarkeit, sowie den Verlust Photosynthese-kompetenter Chloroplasten in gebleichten Sektoren. Diese Defekte werden dadurch verursacht, dass die Plastid-Entwicklung in einem frühen Stadium blockiert wird. Dies führt zu defekten Plastiden, die keine kanonischen internen Strukturen, einschließlich Grana, bilden können. Die gestörte Plastid-Entwicklung ist ein Resultat fehlerhafter Prozessierung ribosomaler RNAs und dem daraus folgenden Verlust plastidärer Transkription und Translation. Wenn ERL1 runterreguliert oder überexprimiert ist, akkumulieren 3‘-elongierte 5S rRNA-Moleküle, was Störungen in der Produktion der Ribosomen hervorruft. Die Reifung der 5S rRNA ist leit langem als Prozess bekannt, der viele aufeinander folgende endonukleolytische Spaltungen sowie exonukleolytische Rezessionen beinhaltet. Bis dato war die Gesamtheit der Exonukleasen während dieser Reifung jedoch nur lückenhaft bekannt. Die Ergebnisse dieser Arbeit zeigen, dass ERL1 eine wichtige Rolle in der Plastid-Entwicklung spielt, indem ERL1 den finalen Reifungsschritt des 5S rRNA 3‘-Endes katalysiert.

Effect of management strategies on the performance of ruminant livestock production systems and the safety of plant and animal products in the city of Sikasso, Mali

The regional population growth in West Africa, and especially its urban centers, will bring about new and critical challenges for urban development policy, especially in terms of ensuring food security and providing employment for the growing population. (Peri-) urban livestock and vegetable production systems, which can contribute significantly to these endeavours, are limited by various constraints, amongst them limited access to expensive production factors and their (in)efficient use. To achieve sustainable production systems with low consumer health risks, that can meet the urban increased demand, this doctoral thesis determined nutrient use efficiencies in representative (peri-) urban livestock production systems in three West African cities, and investigated potential health risks for consumers ensuing from there. The field study, which was conducted during July 2007 to December 2009, undertook a comparative analysis of (peri-) urban livestock production strategies across 210 livestock keeping households (HH) in the three West African cities of Kano/Nigeria (84 HH), Bobo Dioulasso/Burkina Faso (63 HH) and Sikasso/Mali (63 HH). These livestock enterprises were belonging to the following three farm types: commercial gardening plus field crops and livestock (cGCL; 88 HH), commercial livestock plus subsistence field cropping (cLsC; 109 HH) and commercial gardening plus semi-commercial livestock (cGscL; 13 HH) which had been classified in a preceding study; they represented the diversity of (peri-) urban livestock production systems in West Africa. In the study on the efficiency of ruminant livestock production, lactating cowsand sheep herd units were differentiated based on whether feed supplements were offered to the animals at the homestead (Go: grazing only; Gsf: mainly grazing plus some supplement feeding). Inflows and outflows of nutrients were quantified in these herds during 18 months, and the effects of seasonal variations in nutrient availability on animals’ productivity and reproductive performance was determined in Sikasso. To assess the safety of animal products and vegetables, contamination sources of irrigated lettuce and milk with microbiological contaminants, and of tomato and cabbage with pesticide residues in (peri-) urban agriculture systems of Bobo Dioulasso and Sikasso were characterized at three occasions in 2009. Samples of irrigation water, organic fertilizer and ix lettuce were collected in 6 gardens, and samples of cabbage and tomato in 12 gardens; raw and curdled milk were sampled in 6 dairy herds. Information on health risks for consumers of such foodstuffs was obtained from 11 health centers in Sikasso. In (peri-) urban livestock production systems, sheep and goats dominated (P<0.001) in Kano compared to Bobo Dioulasso and Sikasso, while cattle and poultry were more frequent (P<0.001) in Bobo Dioulasso and Sikasso than in Kano. Across cities, ruminant feeding relied on grazing and homestead supplementation with fresh grasses, crop residues, cereal brans and cotton seed cake; cereal grains and brans were the major ingredients of poultry feeds. There was little association of gardens and livestock; likewise field cropping and livestock were rarely integrated. No relation existed between the education of the HH head and the adoption of improved management practices (P>0.05), but the proportion of HH heads with a long-term experience in (peri-) urban agriculture was higher in Kano and in Bobo Dioulasso than in Sikasso (P<0.001). Cattle and sheep fetched highest market prices in Kano; unit prices for goats and chicken were highest in Sikasso. Animal inflow, outflow and dairy herd growth rates were significantly higher (P<0.05) in the Gsf than in the Go cattle herds. Maize bran and cottonseed expeller were the main feeds offered to Gsf cows as dry-season supplement, while Gsf sheep received maize bran, fresh grasses and cowpea pods. The short periodic transhumance of Go dairy cows help them maintaining their live weight, whereas Gsf cows lost weight during the dry season despite supplement feeding at a rate of 1506 g dry matter per cow and day, resulting in low productivity and reproductive performance. The daily live weight gains of calves and lambs, respectively, were low and not significantly different between the Go and the Gsf system. However, the average live weight gains of lambs were significantly higher in the dry season (P<0.05) than in the rainy season because of the high pressure of gastrointestinal parasites and of Trypanosoma sp. In consequence, 47% of the sheep leaving the Go and Gsf herds died due to diseases during the study period. Thermo-tolerant coliforms and Escherichia coli contamination levels of irrigation water significantly exceeded WHO recommendations for the unrestricted irrigation of vegetables consumed raw. Microbial contamination levels of lettuce at the farm gate and the market place in Bobo Dioulasso and at the farm gate in Sikasso were higher than at the market place in Sikasso (P<0.05). Pesticide residues were detected in only one cabbage and one tomato sample and were below the maximum residue limit for consumption. Counts of thermo-tolerant coliforms and Escherichia coli were higher in curdled than in raw milk (P<0.05). From 2006 to x 2009, cases of diarrhea/vomiting and typhoid fever had increased by 11% and 48%, respectively, in Sikasso. For ensuring economically successful and ecologically viable (peri-) urban livestock husbandry and food safety of (peri-) urban foodstuffs of animal and plant origin, the dissemination and adoption of improved feeding practices, livestock healthcare and dung management are key. In addition, measures fostering the safety of animal products and vegetables including the appropriate use of wastewater in (peri-) urban agriculture, restriction to approve vegetable pesticides and the respect of their latency periods, and passing and enforcement of safety laws is required. Finally, the incorporation of environmentally sound (peri-) urban agriculture in urban planning by policy makers, public and private extension agencies and the urban farmers themselves is of utmost importance. To enable an efficient (peri-) urban livestock production in the future, research should concentrate on cost-effective feeding systems that allow meeting the animals’ requirement for production and reproduction. Thereby focus should be laid on the use of crop-residues and leguminous forages. The improvement of the milk production potential through crossbreeding of local cattle breeds with exotic breeds known for their high milk yield might be an accompanying option, but it needs careful supervision to prevent the loss of the local trypanotolerant purebreds.

Effects of plant diversity on bioenergy parameters in grassland biomass

Extensive grassland biomass for bioenergy production has long been subject of scientific research. The possibility of combining nature conservation goals with a profitable management while reducing competition with food production has created a strong interest in this topic. However, the botanical composition will play a key role for solid fuel quality of grassland biomass and will have effects on the combustion process by potentially causing corrosion, emission and slagging. On the other hand, botanical composition will affect anaerobic digestibility and thereby the biogas potential. In this thesis aboveground biomass from the Jena-Experiment plots was harvested in 2008 and 2009 and analysed for the most relevant chemical constituents effecting fuel quality and anaerobic digestibility. Regarding combustion, the following parameters were of main focus: higher heating value (HHV), gross energy yield (GE), ash content, ash softening temperature (AST), K, Ca, Mg, N, Cl and S content. For biogas production the following parameters were investigated: substrate specific methane yield (CH4 sub), area specific methane yield (CH4 area), crude fibre (CF), crude protein (CP), crude lipid (CL) and nitrogen-free extract (NfE). Furthermore, an improvement of the fuel quality was investigated through applying the Integrated generation of solid Fuel and Biogas from Biomass (IFBB) procedure. Through the specific setup of the Jena-Experiment it was possible to outline the changes of these parameters along two diversity gradients: (i) species richness (SR; 1 to 60 species) and (ii) functional group (grasses, legumes, small herbs and tall herbs) presence. This was a novel approach on investigating the bioenergy characteristic of extensive grassland biomass and gave detailed insight in the sward-composition¬ - bioenergy relations such as: (i) the most relevant SR effect was the increase of energy yield for both combustion (annual GE increased by 26% from SR8→16 and by 65% from SR8→60) and anaerobic digestion (annual CH4 area increased by 22% from SR8→16 and by 49% from SR8→60) through a strong interaction of SR with biomass yield; (ii) legumes play a key role for the utilization of grassland biomass for energy production as they increase the energy content of the substrate (HHV and CH4 sub) and the energy yield (GE and CH4 area); (iii) combustion is the conversion technique that will yield the highest energy output but requires an improvement of the solid fuel quality in order to reduce the risk of corrosion, emission and slagging related problems. This was achieved through applying the IFBB-procedure, with reductions in ash (by 23%), N (28%), K (85%), Cl (56%) and S (59%) and equal levels of concentrations along the SR gradient.

Alternative plant protein sources for pigs and chickens in the tropics – nutritional value and constraints: a review

In the tropics, a large number of smallholder farms contribute significantly to food security by raising pigs and poultry for domestic consumption and for sale on local markets. The high cost and, sometimes, the lack of availability of commercial protein supplements is one of the main limitations to efficient animal production by smallholders. Locally-grown forages and grain legumes offer ecological benefits such as nitrogen fixation, soil improvement, and erosion control which contribute to improve cropping efficiency. Besides these agronomical assets, they can be used as animal feeds in mixed farming systems. In this paper we review options to include locally-grown forages and grain legumes as alternative protein sources in the diets of pigs and poultry in order to reduce farmers’ dependence on externally-purchased protein concentrates. The potential nutritive value of a wide range of forages and grain legumes is presented and discussed. The influence of dietary fibre and plant secondary metabolites contents and their antinutritive consequences on feed intake, digestive processes and animal performances are considered according to the varying composition in those compounds of the different plant species and cultivars covered in this review. Finally, methods to overcome the antinutritive attributes of the plant secondary metabolites using heat, chemical or biological treatment are reviewed regarding their efficiency and their suitability in low input farming systems.

Effects of transformation processes in 'jubraka' agroforestry systems of the Nuba Mountains, Sudan, on plant diversity

Deutsche Forschungsgemeinschaft

Influence of pre-plant densities of Meloidogyne incognita on growth and root infestation of spinach (Spinacia oleracea L.) (Amaranthaceae) – an important dimension towards enhancing crop production

Vegetables represent a main source of micro-nutrients which can improve the health status of malnourished poor in the world. Spinach (Spinacia oleracea L.) is a popular leafy vegetable in many countries which is rich with several important micro-nutrients. Thus, consuming Spinach helps to overcome micro-nutrient deficiencies. Pests and pathogens act as major yield constraints in food production. Root-knot nematodes, Meloidogyne species, constitute a large group of highly destructive plant pests. Spinach is found to be highly susceptible for these nematode attacks. Though agricultural production has largely benefited from modern technologies and innovations, some important dimensions which can minimize the yield losses have been neglected by most of the growers. Pre-plant or initial nematode density in soil is a crucial biotic factor which is directly responsible for crop losses. Hence, information on preplant nematode densities and the corresponding damage is of vital importance to develop successful control procedures to enhance crop production. In the present study, effect of seven initial densities of M. incognita, i.e., 156, 312, 625, 1250, 2,500, 5,000 and 10,000 infective juveniles (IJs)/plant (equivalent to 1000cm3 soil) on the growth and root infestation on potted spinach plants was determined in a screen house. In order to ensure a high accuracy, root infestation was ascertained by the number of galls formed, the percentage galled-length of feeder roots and galled-feeder roots, and egg production, per plant. Fifty days post-inoculation, shoot length and weight, and root length were suppressed at the lowest IJs density. However, the pathogenic effect was pronounced at the highest density at which 43%, 46% and 45% reduction in shoot length and weight, and root length, respectively, was recorded. The highest reduction in root weight (26%) was detected at the second highest density. The Number of galls and percentage galled-length of feeder roots/per plant showed significant progressive increase across the increasing IJs density with the highest mean value of 432.3 and 54%, respectively. The two shoot growth parameters and root length showed significant inverse relationship with the increasing gall formation. Moreover, the shoot and root length were shown to be mutually dependent on each other. Suppression of shoot growth of spinach greatly affects the grower’s economy. Hence, control measures are essentially needed to ensure a better production of spinach via reducing the pre-plant density below the level of 0.156 IJs/cm3.

Effects of shoot pruning and inflorescence thinning on plant growth, yield and fruit quality of greenhouse tomatoes in a tropical climate

The combined effects of shoot pruning (one or two stems) and inflorescence thinning (five or ten flowers per inflorescence) on greenhouse tomato yield and fruit quality were studied during the dry season (DS) and rainy season (RS) in Central Thailand. Poor fruit set, development of undersized (mostly parthenocarpic) fruits, as well as the physiological disorders blossom-end rot (BER) and fruit cracking (FC) turned out to be the prevailing causes deteriorating fruit yield and quality. The proportion of marketable fruits was less than 10% in the RS and around 65% in the DS. In both seasons, total yield was significantly increased when plants were cultivated with two stems, resulting in higher marketable yields only in the DS. While the fraction of undersized fruits was increased in both seasons when plants were grown with a secondary stem, the proportions of BER and FC were significantly reduced. Restricting the number of flowers per inflorescence invariably resulted in reduced total yield. However, in neither season did fruit load considerably affect quantity or proportion of the marketable yield fraction. Inflorescence thinning tended to promote BER and FC, an effect which was only significant for BER in the RS. In conclusion, for greenhouse tomato production under climate conditions as they are prevalent in Central Thailand, the cultivation with two stems appears to be highly recommendable whereas the measures to control fruit load tested in this study did not proof to be advisable.

Privatization in Colombia: a plant performance analysis

El artículo describe el programa de privatización en el sector real de la economía Colombiana durante los años noventa y ubica esta política en un contexto de desregulación de mercados y promoción de la inversión privada en la oferta de infraestructura publica y servicios públicos domiciliarios. El articulo evalúa el programa de privatización en los sectores manufacturero y de generación de energía eléctrica. Se hacen mediciones ex - post y análisis econométrico del desempeño de las firmas privatizadas. En el sector manufacturero la muestra analizada esta compuesta por 30 firmas manufactureras de gran tamaño, donde el Instituto de Fomento Industrial era socio fundador. Los principales resultados sugieren que estas firmas mantuvieron un comportamiento pro - cíclico relativo a su principal competidor privado y desestimaron planes drásticos de reestructuración operativa. Para el grupo de firmas de generación de energía el articulo estudia el impacto de la reforma regulatoria en el proceso de entrada al mercado, estructura de propiedad, competencia de mercado y eficiencia productiva. La medición de eficiencia productiva usa la técnica de Data Envelope Análisis para 33 plantas que representan el 85% de la capacidad instalada en la generación térmica de energía eléctrica. La muestra esta compuesta por plantas que estaban en funcionamiento antes de la reforma y las entrantes que comenzaron su operación comercial después de la reforma. Los resultados sugieren que los niveles de evidencia en la generación térmica a mejorado después de la reforma y que la política regulatoria ha tenido un efecto positivo en la eficiencia productiva.

Genotypic comparison of Pantoea agglomerans plant and clinical strains

Pantoea agglomerans strains are among the most promising biocontrol agents for a variety of bacterial and fungal plant diseases, particularly fire blight of apple and pear. However, commercial registration of P. agglomerans biocontrol products is hampered because this species is currently listed as a biosafety level 2 (BL2) organism due to clinical reports as an opportunistic human pathogen. This study compares plant-origin and clinical strains in a search for discriminating genotypic/phenotypic markers using multi-locus phylogenetic analysis and fluorescent amplified fragment length polymorphisms (fAFLP) fingerprinting. Results: Majority of the clinical isolates from culture collections were found to be improperly designated as P. agglomerans after sequence analysis. The frequent taxonomic rearrangements underwent by the Enterobacter agglomerans/Erwinia herbicola complex may be a major problem in assessing clinical associations within P. agglomerans. In the P. agglomerans sensu stricto (in the stricter sense) group, there was no discrete clustering of clinical/biocontrol strains and no marker was identified that was uniquely associated to clinical strains. A putative biocontrol-specific fAFLP marker was identified only in biocontrol strains. The partial ORF located in this band corresponded to an ABC transporter that was found in all P. agglomerans strains. Conclusion: Taxonomic mischaracterization was identified as a major problem with P. agglomerans, and current techniques removed a majority of clinical strains from this species. Although clear discrimination between P. agglomerans plant and clinical strains was not obtained with phylogenetic analysis, a single marker characteristic of biocontrol strains was identified which may be of use in strain biosafety determinations. In addition, the lack of Koch's postulate fulfilment, rare retention of clinical strains for subsequent confirmation, and the polymicrobial nature of P. agglomerans clinical reports should be considered in biosafety assessment of beneficial strains in this species

Control and optimization of an SBR for nitrogen removal: from model calibration to plant operation

En aquesta tesis s'ha desenvolupat un sistema de control capaç d'optimitzar el funcionament dels Reactors Discontinus Seqüencials dins el camp de l'eliminació de matèria orgànica i nitrogen de les aigües residuals. El sistema de control permet ajustar en línia la durada de les etapes de reacció a partir de mesures directes o indirectes de sondes. En una primera etapa de la tesis s'ha estudiat la calibració de models matemàtics que permeten realitzar fàcilment provatures de diferents estratègies de control. A partir de l'anàlisis de dades històriques s'han plantejat diferents opcions per controlar l'SBR i les més convenients s'han provat mitjançant simulació. Després d'assegurar l'èxit de l'estratègia de control mitjançant simulacions s'ha implementat en una planta semi-industrial. Finalment es planteja l'estructura d'uns sistema supervisor encarregat de controlar el funcionament de l'SBR no només a nivell de fases sinó també a nivell cicle.

Design and synthesis of short antimicrobial peptides for plant protection. Study of their mode of action

Aquesta tesi doctoral està basada en el desenvolupament de nous agents antimicrobians derivats del pèptid híbrid cecropina A-melitina WKLFKKILKVL-NH2 (Pep3) que siguin sostenibles i útils per al control de malalties de plantes. Es van dissenyar i sintetitzar més de 133 anàlegs de Pep3 mitjançant química combinatòria. Es van obtenir anàlegs de Pep3 amb una elevada activitat contra fitopatògens i que presentaven baixa toxicitat. Els millors anàlegs van presentar eficàcies comparables amb pesticides de referència en la prevenció d'infeccions causades per fitopatògens. Es va estudiar el mecanisme d'acció de KKLFKKILKYL-NH2 (BP100) investigant la seva interacció amb models de membrana mitjançant tècniques espectroscòpiques. Es va observar la capacitat de BP100 a induir la permeabilització, la neutralització, i l'agregació de vesícules lipídiques aniòniques a una determinada concentració llindar. Es va deduir una equació que relaciona la CMI d'un pèptid antimicrobià amb la constant de partició i la concentració llindar en la membrana.

Synthesis and evaluation of cyclic cationic peptides as antimicrobial agents for use in plant protection

Aquesta tesi doctoral se centra en l'estudi de l'aplicació de pèptids antimicrobians en la lluita contra agents patògens de cultius de plantes d'interès econòmic.L'estratègia sintètica s'ha portat a terme utilitzant metodologies convencionals de síntesi de pèptids en fase sòlida com l'estratègia tridimensional ortogonal Fmoc/tBut/Allyl. Ha calgut fer la recerca de les condicions òptimes per a l'eliminació del grup Allyl i la ciclació. D'entre els pèptids cíclics de 4-10 aminoacids sintetitzats, el decapèptid c(Lys-Leu-Lys-Leu-Lys-Phe-Lys-Lys-Leu-Gln) ha resultat ésser el més efectiu i s'ha pres com a base per al disseny d'una quimioteca de 56 pèptids. Dels resultats obtinguts s'ha sintetitzat una segona quimioteca basada en l'estructura general c(X1-X2-X3-X4-Lys-Phe-Lys-Lys-Leu-Gln) determinada com la que posseix el millor perfil d'activitat. Els pèptids més efectius obtinguts constituixen els primers exemples de pèptids cíclics actius contra E. amylovora i poden ser considerats com a bons candidats pel desenvolupament d'agents antimicrobians efectius en protecció vegetal.