Translocation and Functional Analysis of Pseudomonas savastanoi pv. savastanoi NCPPB 3335 Type III Secretion System Effectors Reveals Two Novel Effector Families of the Pseudomonas syringae Complex

Pseudomonas savastanoi pv. savastanoi NCPPB 3335 causes olive knot disease and is a model pathogen for exploring bacterial infection of woody hosts. The type III secretion system (T3SS) effector repertoire of this strain includes 31 effector candidates plus two novel candidates identified in this study which have not been reported to translocate into plant cells. In this work, we demonstrate the delivery of seven NCPPB 3335 effectors into Nicotiana tabacum leaves, including three proteins from two novel families of the P. syringae complex effector super-repertoire (HopBK and HopBL), one of which comprises two proteins (HopBL1 and HopBL2) that harbor a SUMO protease domain. When delivered by P. fluorescens heterologously expressing a P. syringae T3SS, all seven effectors were found to suppress the production of defense-associated reactive oxygen species. Moreover, six of these effectors, including the truncated versions of HopAA1 and HopAZ1 encoded by NCPPB 3335, suppressed callose deposition. The expression of HopAZ1 and HopBL1 by functionally effectorless P. syringae pv. tomato DC3000D28E inhibited the hypersensitive response in tobacco and, additionally, expression of HopBL2 by this strain significantly increased its competitiveness in N. benthamiana. DNA sequences encoding HopBL1 and HopBL2 were uniquely detected in a collection of 31 P. savastanoi pv. savastanoi strains and other P. syringae strains isolated from woody hosts, suggesting a relevant role of these two effectors in bacterial interactions with olive and other woody plants.

Vehicle to vehicle energy exchange in smart grid applications

This paper presents a novel vehicle to vehicle energy exchange market (V2VEE) between electric vehicles (EVs) for decreasing the energy cost to be paid by some users whose EVs must be recharged during the day to fulfil their daily scheduled trips and also reducing the impact of charging on the electric grid. EVs with excess of energy in their batteries can transfer this energy among other EVs which need charge during their daily trips. These second type of owners can buy the energy directly to the electric grid or they can buy the energy from other EV at lower price. An aggregator is responsible for collecting all information among vehicles located in the same area at the same time and make possible this energy transfer.

A voltage disturbances prototype for testing electrical generators connected to microgrids

The search for new energy models arises as a necessity to have a sustainable power supply. The inclusion of distributed generation sources (DG) allows to reduce the cost of facilities, increase the security of the grid or alleviate problems of congestion through the redistribution of power flows. In remote microgrids it is needed in a particular way a safe and reliable supply, which can cover the demand for a low cost; due to this, distributed generation is an alternative that is being widely introduced in these grids. But the remote microgrids are especially weak grids because of their small size, low voltage level, reduced network mesh and distribution lines with a high ratio R/X. This ratio affects the coupling between grid voltages and phase shifts, and stability becomes an issue of greater importance than in interconnected systems. To ensure the appropriate behavior of generation sources inserted in remote microgrids -and, in general, any electrical equipment-, it is essential to have devices for testing and certification. These devices must, not only faithfully reproduce disturbances occurring in remote microgrids, but also to behave against the equipment under test (EUT) as a real weak grid. This also makes the device commercially competitive. To meet these objectives and based on the aforementioned, it has been designed, built and tested a voltage disturbances generator, in order to provide a simple, versatile, full and easily scalable device to manufacturers and laboratories in the sector.

A multi-level analysis of innovation diffusion: The case of solar photovoltaic systems in Germany and the disillusion of grid parity

¿Suministrarán las fuentes de energía renovables toda la energía que el mundo necesita algún día? Algunos argumentan que sí, mientras que otros dicen que no. Sin embargo, en algunas regiones del mundo, la producción de electricidad a través de fuentes de energía renovables ya está en una etapa prometedora de desarrollo en la que su costo de generación de electricidad compite con fuentes de electricidad convencionales, como por ejemplo la paridad de red. Este logro ha sido respaldado por el aumento de la eficiencia de la tecnología, la reducción de los costos de producción y, sobre todo, los años de intervenciones políticas de apoyo financiero. La difusión de los sistemas solares fotovoltaicos (PV) en Alemania es un ejemplo relevante. Alemania no sólo es el país líder en términos de capacidad instalada de sistemas fotovoltaicos (PV) en todo el mundo, sino también uno de los países pioneros donde la paridad de red se ha logrado recientemente. No obstante, podría haber una nube en el horizonte. La tasa de difusión ha comenzado a declinar en muchas regiones. Además, las empresas solares locales – que se sabe son importantes impulsores de la difusión – han comenzado a enfrentar dificultades para manejar sus negocios. Estos acontecimientos plantean algunas preguntas importantes: ¿Es ésta una disminución temporal en la difusión? ¿Los adoptantes continuarán instalando sistemas fotovoltaicos? ¿Qué pasa con los modelos de negocio de las empresas solares locales? Con base en el caso de los sistemas fotovoltaicos en Alemania a través de un análisis multinivel y dos revisiones literarias complementarias, esta tesis doctoral extiende el debate proporcionando riqueza múltiple de datos empíricos en un conocimiento de contexto limitado. El primer análisis se basa en la perspectiva del adoptante, que explora el nivel "micro" y el proceso social que subyace a la adopción de los sistemas fotovoltaicos. El segundo análisis es una perspectiva a nivel de empresa, que explora los modelos de negocio de las empresas y sus roles impulsores en la difusión de los sistemas fotovoltaicos. El tercero análisis es una perspectiva regional, la cual explora el nivel "meso", el proceso social que subyace a la adopción de sistemas fotovoltaicos y sus técnicas de modelado. Los resultados incluyen implicaciones tanto para académicos como políticos, no sólo sobre las innovaciones en energía renovable relativas a la paridad de red, sino también, de manera inductiva, sobre las innovaciones ambientales impulsadas por las políticas que logren la competitividad de costes. ABSTRACT Will renewable energy sources supply all of the world energy needs one day? Some argue yes, while others say no. However, in some regions of the world, the electricity production through renewable energy sources is already at a promising stage of development at which their electricity generation costs compete with conventional electricity sources’, i.e., grid parity. This achievement has been underpinned by the increase of technology efficiency, reduction of production costs and, above all, years of policy interventions of providing financial support. The diffusion of solar photovoltaic (PV) systems in Germany is an important frontrunner case in point. Germany is not only the top country in terms of installed PV systems’ capacity worldwide but also one of the pioneer countries where the grid parity has recently been achieved. However, there might be a cloud on the horizon. The diffusion rate has started to decline in many regions. In addition, local solar firms – which are known to be important drivers of diffusion – have started to face difficulties to run their businesses. These developments raise some important questions: Is this a temporary decline on diffusion? Will adopters continue to install PV systems? What about the business models of the local solar firms? Based on the case of PV systems in Germany through a multi-level analysis and two complementary literature reviews, this PhD Dissertation extends the debate by providing multiple wealth of empirical details in a context-limited knowledge. The first analysis is based on the adopter perspective, which explores the “micro” level and the social process underlying the adoption of PV systems. The second one is a firm-level perspective, which explores the business models of firms and their driving roles in diffusion of PV systems. The third one is a regional perspective, which explores the “meso” level, i.e., the social process underlying the adoption of PV systems and its modeling techniques. The results include implications for both scholars and policymakers, not only about renewable energy innovations at grid parity, but also in an inductive manner, about policy-driven environmental innovations that achieve the cost competiveness.

Mecanismos implicados en las etapas iniciales de infección en la cancrosis de los cítricos provocada por Xanthomonas citri subsp. citri

La cancrosis o chancro bacteriano de los cítricos (CBC) causada por Xanthomonas citri subsp. citri (Xcc) y X. fuscans subsp. aurantifolii, afecta a un gran número de especies dentro de la familia de las rutáceas, especialmente cítricos. Esta enfermedad produce graves pérdidas económicas allí donde está presente, principalmente porque la comercialización de cítricos desde las zonas afectadas hacía zonas libres de cancrosis, está sujeta a fuertes medidas cuarentenarias. La cancrosis se encuentra distribuida a nivel mundial pero no se ha localizado ni en la Unión Europea ni en ningún área del Mediterráneo. Se han descrito tres tipos de cancrosis en función de la gama de huésped y de las características fenotípicas y genotípicas de las bacterias que las producen. La más extendida es la cancrosis tipo A producida por Xcc, dentro de la cual se distinguen los subtipos Aw y A*, originarios de Florida y Sudeste Asiático, respectivamente, que de forma natural solo son capaces de producir enfermedad en lima mejicana. En este trabajo se presentan estudios sobre mecanismos implicados en las primeras etapas de la infección, como la quimiotaxis y formación de biopelículas, en la cancrosis de los cítricos. La quimiotaxis es el proceso por el cual las bacterias se dirigen hacia zonas favorables para su supervivencia y desarrollo. Los perfiles quimiotácticos obtenidos frente a distintas fuentes de carbono, así como los estudios en relación al contenido de proteínas aceptoras de grupos metilo (MCPs), permitieron agrupar a las cepas de Xanthomonas estudiadas en este trabajo, de acuerdo a la enfermedad producida y a su gama de huésped. Todas las cepas mostraron quimiotaxis positiva frente a extractos de hoja y apoplasto de diferentes especies, sin embargo, Xcc 306, X. alfalfae subsp. citrumelonis (Xac) y X. campestris pv. campestris (Xc) manifestaron respuestas más específicas frente a extractos de apoplasto de hojas de naranjo dulce, lima y col china, respectivamente. Dicho resultado nos permite asociar el mecanismo de quimiotaxis con la capacidad de las cepas de Xanthomonas para colonizar estos huéspedes de forma específica. Las cepas estudiadas fueron capaces de realizar movimiento tipo swimming, twitching y sliding en distintos medios, siendo el movimiento swimming el único en el que se encontraron diferencias entre las cepas de Xcc con distinta gama de huésped. En este trabajo se ha estudiado además la formación de biopelículas en superficies bióticas y abióticas, un mecanismo importante tanto para la supervivencia en superficie vegetal como para el desarrollo de la infección. Las cepas de Xanthomonas estudiadas fueron capaces de formar biopelículas in vitro, siendo mayor en un medio que simula el apoplasto y que contiene una baja concentración de nutrientes en comparación con medios que contenían alta concentración de nutrientes. La formación de biopelículas en superficie vegetal se encontró relacionada, en las cepas patógenas de cítricos, con la capacidad para infectar un tejido o huésped determinado. Se han caracterizado algunos de los componentes de la matriz extracelular producida por Xcc, que compone hasta un 90% de las bipoelículas. Entre ellos destaca el ADN extracelular, que tiene un papel como adhesina en las primeras etapas de formación de biopelículas y estructural en biopelículas maduras. Además, se han identificado el pilus tipo IV como componente importante en las biopelículas, que también participa en motilidad. Finalmente, se han realizado estudios sobre la expresión de genes implicados en motilidad bacteriana y formación de biopelículas que han confirmado las diferencias existentes entre cepas de Xcc de amplia y limitada gama de huésped, así como el papel que juegan elementos como el pilus tipo IV o el flagelo en estos procesos. ABSTRACT Xanthomonas citri subsp. citri (Xcc) and X. fuscans subsp. aurantifolii are the causal agents of Citrus Bacterial Canker (CBC) which is one of the most important citrus diseases. CBC affects all Citrus species as well as other species from Rutaceae family. CBC produces strong economic losses; furthermore the commercialization of plants and fruits is restricted from infested to citrus canker free areas. The disease is worldwide distributed in tropical and subtropical areas, however it is not present in the European Union. Three types of CBC have been described according to the host range and phenotypic and genotypic characteristics. CBC type A caused by Xcc is he widest distributed. Within CBC A type two subtypes Aw and A* were described from Florida and Iran respectively, both infecting only Mexican lime. Herein mechanisms connected to early events in the citrus bacterial canker disease such as chemotaxis and biofilm formation, were studied. Chemotaxis allows bacteria to move towards the more suitable environments for its survival, host colonization and infection. Studies performed on citrus pathogenic Xanthomonas and X. campestris pv. campestris (Xc), a crucifer pathogen, have shown different chemotactic profiles towards carbon compound as well as different MCPs profile, which clustered strains according to host range and disease caused. Every strain showed positive chemotaxis toward leaf extracts and apoplastic fluids from sweet orange, Mexican lime and Chinese cabbage leaves. However, a more specific response was found for strains Xcc 306, X. alfalfae subsp. citrumelonis and Xc towards sweet orange, Mexican lime and Chinese cabbage apoplastic fluids, respectively. These results relate chemotaxis with the higher ability of those strains to specifically colonize their proper host. Xanthomonas strains studied were able to perform swimming, sliding and twitching motilities. The ability to swim was variable among CBC strains and seemed related to host range. Biofilm formation is an important virulence factor for Xcc because it allows a better survival onto the plant surface as well as facilitates the infection process. The studied Xanthomonas strains were able to form biofilm in vitro, on both nutrient rich and apoplast mimicking media, furthermore the biofilm formation by all the strains was higher in the apoplast mimicking media. The ability to form biofilm in planta by Xcc and Xac strains was dependent of the host and the tissue colonized. The wide host range CBC strain was able to form biofilm onto several citrus leaves and fruits, however the limited host range CBC strain produced biofilm solely onto Mexican lime leaves and fruits. Furthermore Xac strain, which solely infects leaves of young plants, was not able to develop biofilms on fruits. Some components of the extracellular matrix produced by Xcc strains have been characterized. Extracellular DNA acted as an adhesin at the very early stages of biofilm formation and as structural component of mature biofilm for citrus pathogenic Xanthomonas. Furthermore type IV pilus has been identified as a component of the extracellular matrix in biofilm and motility. Transcriptional studies of genes related with biofilm formation and motility have confirmed the differential behavior found among wide and limited host range CBC strains as well as the role of type IV pili and flagellum on those processes.

Integración técnica y económica de energías renovables, en un sistema eléctrico liberalizado, a partir de vectores energéticos

El objetivo de la tesis es estudiar la bondad del almacenamiento de energía en hidrógeno para minorar los desvíos de energía respecto a su previsión de parques eólicos y huertas solares. Para ello se ha partido de datos de energías horarias previstas con 24 h de antelación y la energía real generada. Se ha procedido a dimensionar la planta de hidrógeno, a partir de una modelización de la operación de la misma, teniendo siempre como objetivo la limitación de los desvíos. Posteriormente, se ha procedido a simular la operación de la planta con dos objetivos en mente, uno limitar los desvíos y por otro lado operar la planta como una central de bombeo, generando hidrógeno en horas valle y generando electricidad en horas punta. Las dos simulaciones se han aplicado a tres parques eólicos de diferentes potencias, y a una huerta solar fotovoltaica. Se ha realizado un estudio económico para determinar la viabilidad de las plantas dimensionadas, obteniendo como resultado que no son viables a día de hoy y con la estimación de precios considerada, necesitando disminuir considerablemente los costes, dependiendo fuertemente de la bondad de los métodos de previsión de viento. Por último se ha estudiado la influencia de la disminución de los desvíos generados sobre una red tipo de 30 nudos, obteniendo como resultado, que si bien no disminuyen sensiblemente los extra costes generados en regulación, sí que mejora la penetración de las energías renovables no despachables en la red. Se observa disminuyen los vertidos eólicos cuando se usa la planta de hidrógeno. ABSTRACT The aim of this thesis is to study the benefit of hydrogen energy storage to minimize energy deviations of Wind Power and Solar Photovoltaic (PV) Power Plants compared to its forecast. To achieve this goal, first of all we have started with hourly energy data provided 24 h in advance (scheduled energy), and real generation (measured energy). Secondly, It has been sized the hydrogen plant, from a modeling of its working mode, always keeping the goal in mind of limiting energy imbalances. Subsequently, It have been simulated the plant working mode following two goals, one, to limit energy imbalances and secondly to operate the plant as a pumping power plant, generating hydrogen-in valley hours and generating electricity at peak hours. The two simulations have been applied to three wind power plants with different installed power capacities, and a photovoltaic solar power plant. It has been done an economic analysis in order to determine the viability of this sized plants, turning out not viable plants today with the estimated prices considered, requiring significantly lower costs, depending heavily on the reliability of the Wind Power forecast methods. Finally, It has been studied the influence of decreasing measured imbalances (of energy) in a 30 grid node, resulting that, while it not reduces significantly the extra costs generated by reserve power, it does improve the penetration of non-manageable renewable energy on the grid, by reducing the curtailments of power of these plants.

Motivators for adoption of photovoltaic systems at grid parity: A case study from Southern Germany

In some countries, photovoltaic (PV) technology is at a stage of development at which it can compete with conventional electricity sources in terms of electricity generation costs, i.e., grid parity. A case in point is Germany, where the PV market has reached a mature stage, the policy support has scaled down and the diffusion rate of PV systems has declined. This development raises a fundamental question: what are the motives to adopt PV systems at grid parity? The point of departure for the relevant literature has been on the impact of policy support, adopters and, recently, local solar companies. However, less attention has been paid to the motivators for adoption at grid parity. This paper presents an in-depth analysis of the diffusion of PV systems, explaining the impact of policy measures, adopters and system suppliers. Anchored in an extensive and exploratory case study in Germany, we provide a context-specific explanation to the motivations to adopt PV systems at grid parity.